<?xml version="1.0" encoding="UTF-8"?><!DOCTYPE article PUBLIC "-//NLM//DTD JATS (Z39.96) Journal Publishing DTD v1.2 20190208//EN" "http://jats.nlm.nih.gov/publishing/1.2/JATS-journalpublishing1.dtd"><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" article-type="research-article" dtd-version="1.2" xml:lang="en">
    <front>
        <journal-meta>
            <journal-id journal-id-type="pmc">F1000Research</journal-id>
            <journal-title-group>
                <journal-title>F1000Research</journal-title>
            </journal-title-group>
            <issn pub-type="epub">2046-1402</issn>
            <publisher>
                <publisher-name>F1000 Research Limited</publisher-name>
                <publisher-loc>London, UK</publisher-loc>
            </publisher>
        </journal-meta>
        <article-meta>
            <article-id pub-id-type="doi">10.12688/f1000research.74041.1</article-id>
            <article-categories>
                <subj-group subj-group-type="heading">
                    <subject>Research Article</subject>
                </subj-group>
                <subj-group>
                    <subject>Articles</subject>
                </subj-group>
            </article-categories>
            <title-group>
                <article-title>Rice husk and melaleuca biochar additions reduce soil CH
                    <sub>4</sub> and N
                    <sub>2</sub>O emissions and increase soil organic matter and nutrient availability</article-title>
                <fn-group content-type="pub-status">
                    <fn>
                        <p>[version 1; peer review: 1 approved, 1 approved with reservations]</p>
                    </fn>
                </fn-group>
            </title-group>
            <contrib-group>
                <contrib contrib-type="author" corresp="no">
                    <name>
                        <surname>Tran Sy</surname>
                        <given-names>Nam</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Conceptualization</role>
                    <role content-type="http://credit.niso.org/">Data Curation</role>
                    <role content-type="http://credit.niso.org/">Formal Analysis</role>
                    <role content-type="http://credit.niso.org/">Investigation</role>
                    <role content-type="http://credit.niso.org/">Methodology</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Original Draft Preparation</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Review &amp; Editing</role>
                    <uri content-type="orcid">https://orcid.org/0000-0001-6445-0076</uri>
                    <xref ref-type="aff" rid="a1">1</xref>
                </contrib>
                <contrib contrib-type="author" corresp="yes">
                    <name>
                        <surname>Huynh Van</surname>
                        <given-names>Thao</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Conceptualization</role>
                    <role content-type="http://credit.niso.org/">Data Curation</role>
                    <role content-type="http://credit.niso.org/">Formal Analysis</role>
                    <role content-type="http://credit.niso.org/">Investigation</role>
                    <role content-type="http://credit.niso.org/">Methodology</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Original Draft Preparation</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Review &amp; Editing</role>
                    <uri content-type="orcid">https://orcid.org/0000-0001-6582-9061</uri>
                    <xref ref-type="corresp" rid="c1">a</xref>
                    <xref ref-type="aff" rid="a1">1</xref>
                </contrib>
                <contrib contrib-type="author" corresp="no">
                    <name>
                        <surname>Chiem</surname>
                        <given-names>Nguyen Huu</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Project Administration</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Review &amp; Editing</role>
                    <xref ref-type="aff" rid="a1">1</xref>
                </contrib>
                <contrib contrib-type="author" corresp="no">
                    <name>
                        <surname>Nguyen Van</surname>
                        <given-names>Cong</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Conceptualization</role>
                    <role content-type="http://credit.niso.org/">Supervision</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Review &amp; Editing</role>
                    <xref ref-type="aff" rid="a1">1</xref>
                </contrib>
                <contrib contrib-type="author" corresp="no">
                    <name>
                        <surname>Mitsunori</surname>
                        <given-names>Tarao</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Supervision</role>
                    <role content-type="http://credit.niso.org/">Validation</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Review &amp; Editing</role>
                    <xref ref-type="aff" rid="a2">2</xref>
                </contrib>
                <aff id="a1">
                    <label>1</label>College of Environment and Natural resources, 3/2 street, Can Tho University, Can Tho city, 900000, Vietnam</aff>
                <aff id="a2">
                    <label>2</label>Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo 183-8506, 183-8506, Japan</aff>
            </contrib-group>
            <author-notes>
                <corresp id="c1">
                    <label>a</label>
                    <email xlink:href="mailto:hvthao@ctu.edu.vn">hvthao@ctu.edu.vn</email>
                </corresp>
                <fn fn-type="conflict">
                    <p>No competing interests were disclosed.</p>
                </fn>
            </author-notes>
            <pub-date pub-type="epub">
                <day>8</day>
                <month>11</month>
                <year>2021</year>
            </pub-date>
            <pub-date pub-type="collection">
                <year>2021</year>
            </pub-date>
            <volume>10</volume>
            <elocation-id>1128</elocation-id>
            <history>
                <date date-type="accepted">
                    <day>1</day>
                    <month>11</month>
                    <year>2021</year>
                </date>
            </history>
            <permissions>
                <copyright-statement>Copyright: &#x00a9; 2021 Tran Sy N et al.</copyright-statement>
                <copyright-year>2021</copyright-year>
                <license xlink:href="https://creativecommons.org/licenses/by/4.0/">
                    <license-p>This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
                </license>
            </permissions>
            <self-uri content-type="pdf" xlink:href="https://f1000research.com/articles/10-1128/pdf"/>
            <abstract>
                <p>
                    <bold>Background</bold>: Biochar is a promising material in mitigating greenhouse gases (GHGs) emissions from paddy fields due to its remarkable structural properties. Rice husk biochar (RhB) and melaleuca biochar (MB) are amendment materials that could be used to potentially reduce emissions in the Vietnamese Mekong Delta (VMD). However, their effects on CH
                    <sub>4</sub> and N
                    <sub>2</sub>O emissions and soil under local water management and conventional rice cultivation have not been thoroughly investigated.</p>
                <p>
                    <bold>Methods</bold>: We conducted a field experiment using biochar additions to the topsoil layer (0-20 cm). Five treatments comprising 0 t ha
                    <sup>&#x2212;1</sup> (CT0); 5 t ha
                    <sup>&#x2212;1</sup> (RhB5) and 10 t ha
                    <sup>&#x2212;1</sup> (RhB10), and 5 t ha
                    <sup>&#x2212;1</sup> (MB5) and 10 t ha
                    <sup>&#x2212;1</sup> (MB10) were designed plot-by-plot (20 m
                    <sup>2</sup>) in triplicates.</p>
                <p>
                    <bold>Results</bold>: The results showed that biochar application from 5 to 10 t ha
                    <sup>&#x2212;1</sup> significantly decreased cumulative CH
                    <sub>4</sub> (24.2 &#x2013; 28.0%, RhB; 22.0 &#x2013; 14.1%, MB) and N
                    <sub>2</sub>O (25.6 &#x2013; 41.0%, RhB; 38.4 &#x2013; 56.4%, MB) fluxes without a reduction in grain yield. Increasing the biochar application rate further did not decrease significantly total CH
                    <sub>4</sub> and N
                    <sub>2</sub>O fluxes but was seen to significantly reduce the global warming potential (GWP) and yield-scale GWP in the RhB treatments. Biochar application improved soil Eh but had no effects on soil pH. Whereas CH
                    <sub>4</sub> flux correlated negatively with soil Eh (
                    <italic toggle="yes">P &lt;</italic> 0.001; 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.552, RhB; 
                    <italic toggle="yes">P &lt;</italic> 0.001; 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.502, MB). The soil physicochemical properties of bulk density, porosity, organic matter, and anaerobically mineralized N were significantly improved in biochar-amended treatments, while available P also slightly increased.</p>
                <p>
                    <bold>Conclusions</bold>: Biochar supplementation significantly reduced CH
                    <sub>4</sub> and N
                    <sub>2</sub>O fluxes and improved soil mineralization and physiochemical properties toward beneficial for rice plant. The results suggest that the optimal combination of biochar-application rates and effective water-irrigation techniques for soil types in the MD should be further studied in future works.</p>
            </abstract>
            <kwd-group kwd-group-type="author">
                <kwd>Biochar amendment</kwd>
                <kwd>conventional rice farming</kwd>
                <kwd>greenhouse gas emissions</kwd>
                <kwd>melaleuca biochar</kwd>
                <kwd>rice-husk biochar</kwd>
                <kwd>soil fertility</kwd>
            </kwd-group>
            <funding-group>
                <award-group id="fund-1">
                    <funding-source>This study was funded by the Can Tho University Improvement Project VN14-P6, Japanese ODA loan. The authors declare no competing financial interests.</funding-source>
                    <award-id>VN14-P6</award-id>
                </award-group>
                <funding-statement>This study is funded in part by the Can Tho University Improvement Project VN14-P6, supported by a Japanese ODA loan. The authors declare no competing financial interests.</funding-statement>
            </funding-group>
        </article-meta>
    </front>
    <body>
        <sec id="sec1" sec-type="intro">
            <title>Introduction</title>
            <p>In Vietnam, the agricultural sector contributes approximately 30% of national greenhouse gases (GHGs) emissions (MORNE, 2017). For rice cultivation, paddy fields are the primary source of GHGs emissions (
                <xref ref-type="bibr" rid="ref33">Nan 
                    <italic toggle="yes">et al.</italic>, 2020</xref>; 
                <xref ref-type="bibr" rid="ref43">Shinoda 
                    <italic toggle="yes">et al.</italic>, 2019</xref>), accounting for 50% of the sub-sectors in agricultural production and roughly 14.6% of national GHG emissions in Vietnam (
                <xref ref-type="bibr" rid="ref32">MONRE, 2017</xref>). According to 
                <xref ref-type="bibr" rid="ref9">NDC (2020)</xref>, Vietnam is committed to reducing 8% of total national GHGs emissions from domestic resources by 2030. Management and technological strategies will play a vital role in reducing the total carbon footprint. Biochar is a carbonized biomass product produced from thermochemical conversion of organic materials under oxygen-limited conditions (
                <xref ref-type="bibr" rid="ref27">Lohri 
                    <italic toggle="yes">et al.</italic>, 2016</xref>; 
                <xref ref-type="bibr" rid="ref63">Wu 
                    <italic toggle="yes">et al.</italic>, 2012</xref>; 
                <xref ref-type="bibr" rid="ref61">Waqas 
                    <italic toggle="yes">et al.</italic>, 2018</xref>). Biochar applications have been noted as one of the most promising approaches for reducing GHGs emissions from rice production (
                <xref ref-type="bibr" rid="ref24">Koyama 
                    <italic toggle="yes">et al</italic>., 2015</xref>; 
                <xref ref-type="bibr" rid="ref64">Wu 
                    <italic toggle="yes">et al.</italic>, 2019a</xref>; 
                <xref ref-type="bibr" rid="ref34">
                    <italic toggle="yes">Nan et al.</italic>, 2021</xref>), and IPCC recently recommended the method (
                <xref ref-type="bibr" rid="ref21">Ji 
                    <italic toggle="yes">et al.</italic>, 2020</xref>). Previous studies have demonstrated that biochar incorporated into soil paddy fields positively rehabilitated soil properties such as pH neutralization, cation exchange capacity (CEC), and buffering capability, soil organic materials (SOM), and nitrogen storage (
                <xref ref-type="bibr" rid="ref41">Qin 
                    <italic toggle="yes">et al.</italic>, 2016</xref>; 
                <xref ref-type="bibr" rid="ref28">Luo 
                    <italic toggle="yes">et al.</italic>, 2020</xref>); improved plant available water, microporosity, and soil aggregate stability, and decreased bulk density (
                <xref ref-type="bibr" rid="ref5">Burrell 
                    <italic toggle="yes">et al.</italic>, 2016</xref>); effected on soil functions and fertility (
                <xref ref-type="bibr" rid="ref17">Giagnoni 
                    <italic toggle="yes">et al.</italic>, 2019</xref>; 
                <xref ref-type="bibr" rid="ref44">Siedt 
                    <italic toggle="yes">et al.</italic>, 2021</xref>); and ameliorated nutrient availability of carbon (C), nitrogen (N), phosphorus (P), potassium (K), magnesium (Mg) and Calcium (Ca) (
                <xref ref-type="bibr" rid="ref25">Li 
                    <italic toggle="yes">et al.</italic>, 2019</xref>). Furthermore, biochar forms a great habitat for different microorganisms via providing macro-, meso- and micropores (
                <xref ref-type="bibr" rid="ref40">Palansooriya 
                    <italic toggle="yes">et al.</italic>, 2019</xref>; 
                <xref ref-type="bibr" rid="ref64">Wu 
                    <italic toggle="yes">et al.</italic>, 2019a</xref>), supports microbial communities by providing labile C substrates for degradation (
                <xref ref-type="bibr" rid="ref45">Smith 
                    <italic toggle="yes">et al.</italic>, 2010</xref>), stimulating biodiversity and abundance of methanotrophic microbes (
                <xref ref-type="bibr" rid="ref41">Qin 
                    <italic toggle="yes">et al.</italic>, 2016</xref>). Moreover, the addition of biochar to the soil reduces GHGs emissions (
                <xref ref-type="bibr" rid="ref47">Spokas and Reicosky, 2009</xref>; 
                <xref ref-type="bibr" rid="ref24">Koyama 
                    <italic toggle="yes">et al.</italic>, 2015</xref>; 
                <xref ref-type="bibr" rid="ref33">Nan 
                    <italic toggle="yes">et al.</italic>, 2020</xref>; 
                <xref ref-type="bibr" rid="ref20">Huang 
                    <italic toggle="yes">et al.</italic>, 2019</xref>) and increases rice yield under different favorable conditions (
                <xref ref-type="bibr" rid="ref68">Yang 
                    <italic toggle="yes">et al.</italic>, 2019</xref>; 
                <xref ref-type="bibr" rid="ref39">Paiman and Effendy, 2020</xref>).</p>
            <p>In the Mekong Delta (MD), melaleuca is an abundantly available hard firewood resource, accounting for 176,295 ha (
                <xref ref-type="bibr" rid="ref18">GIZ, 2009</xref>); the wood reserve of melaleuca is estimated at 13 million m
                <sup>3</sup>. In addition, rice husk is known as a by-product of rice production, accounting for 20% of rice yields (
                <xref ref-type="bibr" rid="ref10">Chungsangunsit 
                    <italic toggle="yes">et al</italic>., 2009</xref>). It is estimated that VMD annually produces around 1.9 million tons of rice husk (
                <xref ref-type="bibr" rid="ref46">Son 
                    <italic toggle="yes">et al.</italic>, 2017</xref>). Biomass (hardwood and crop residues) are often used as typical feedstock for making biochar pyrolysis owing to their multiple-porous structure (
                <xref ref-type="bibr" rid="ref36">Nguyen 
                    <italic toggle="yes">et al.</italic>, 2018</xref>; 
                <xref ref-type="bibr" rid="ref33">Nan 
                    <italic toggle="yes">et al.</italic>, 2020</xref>), which facilitates the multifunctional purposes of soil amendment and pollutant remediation. Therefore, both melaleuca and rice husk could be used to produce biochar, which is then applied to rice paddy fields as a GHGs emission reduction strategy. Although previous studies have demonstrated the effectiveness of biochar incorporation on reducing GHGs emissions, little attention has been paid to the quantitative variation of rice husk biochar (RhB) and melaleuca biochar (MB) on GHGs emissions and soil improvement in VMD lowland conditions. Moreover, the majority of previous studies exclusively emphasize CH
                <sub>4</sub> and N
                <sub>2</sub>O emissions on water practices by controlled irrigation, and alternative wetting and drying, and midseason drainage (
                <xref ref-type="bibr" rid="ref68">Yang 
                    <italic toggle="yes">et al.</italic> 2019</xref>, 
                <xref ref-type="bibr" rid="ref48">Sriphirom 
                    <italic toggle="yes">et al.</italic> 2020</xref>, 
                <xref ref-type="bibr" rid="ref52">Uno 
                    <italic toggle="yes">et al.</italic> 2021</xref>), while atypical water irrigation regime has not been thoroughly elucidated.</p>
            <p>Thus, we aimed (i) to elucidate the CH
                <sub>4</sub> and N
                <sub>2</sub>O emissions and global warming potential (GWP) from the incorporation of RhB and MB into the paddy field soils under locally typical water management regimes in the VMD, and (ii) to determine the effects of RhB and MB amendments on soil physicochemical properties. We, therefore, conducted a field experiment with a variety of RhB and MB amendment amounts under conventional farming practices. Our field experimentation confirmed that RhB and MB application to rice paddy fields was feasible in reducing GHGs emissions. Simultaneously, biochar application improved soil availability of SOM and anaerobically mineralized N.</p>
        </sec>
        <sec id="sec2" sec-type="methods">
            <title>Methods</title>
            <sec id="sec3">
                <title>Site description</title>
                <p>A field experiment was carried out on a typical smallholding farmer's paddy field in Thoi An Dong Village, Can Tho city, Vietnam (10&#x00b0;3&#x2032;44&#x2033;N, 105&#x00b0;41&#x2032;55&#x2033;E). The study area was located in the center of the Mekong Delta, Vietnam, which is a tropical area influenced by the monsoon climate zone, with measured mean annual rainfall (2,088.4 mm), air temperature (27.5 &#x2013; 27.5 &#x00b0;C), humidity (78.0 &#x2013; 86.0%), sunshine (2,467.4 &#x2013; 2,695.4 hours) in the period from 2015-2019 (
                    <xref ref-type="bibr" rid="ref12">DONRE, 2020</xref>). The precipitation and temperature during the experiment were recorded by a weather station placed at the farmer's house (~150 m from the field experiment). The soil was classified as Thionic Glycesol (International Union of Soil Sciences (IUSS) working group World Reference Base (WRB), 2015) (
                    <xref ref-type="bibr" rid="ref11">Dong 
                        <italic toggle="yes">et al.</italic>, 2012</xref>, 
                    <xref ref-type="bibr" rid="ref29">Minamikawa 
                        <italic toggle="yes">et al.,</italic> 2021</xref>). The elementary properties were (0-20 cm depth) as follows: pH (H
                    <sub>2</sub>O), 5.41; EC, 0.9 mS cm
                    <sup>&#x2212;1</sup>; bulk density, 0.92 g.cm
                    <sup>&#x2212;3</sup>, silty clay texture (59.3% clay, 39.5% silt, 1.2% sand); organic matter, 87 g kg
                    <sup>&#x2212;1</sup>; total N, 4.21 g kg
                    <sup>&#x2212;1</sup>; cation exchange capacity (CEC), 37.4 meq 100 g
                    <sup>&#x2212;1</sup>; exchangeable K, 0.54 meq 100 g
                    <sup>&#x2212;1</sup>, exchangeable Mg, 5.47 meq 100 g
                    <sup>&#x2212;1</sup>; exchangeable Ca, 10.5 meq 100 g
                    <sup>&#x2212;1</sup>; and total C, 40.76 g kg
                    <sup>&#x2212;1</sup>.</p>
            </sec>
            <sec id="sec4">
                <title>Biochar preparation</title>
                <p>RhB was made on-site using a simple semi-industrial pyrolysis batch method (
                    <xref ref-type="bibr" rid="ref37">Oikawa 
                        <italic toggle="yes">et al</italic>., 2016</xref>). Here a short iron bar was to set onto the ground. A stainless chimney pipe 1.5m long was vertically erected to the bar using wire. The pipe was kept at a 10-cm distance from the ground to release smoke generated during the pyrolysis process. Embers were placed adjacent to the bar to kick off the carbonization process. Then, rice husk was poured around the bar according to a coniform shape with 1.5 m height and 1.5 m diameter. RhB was generated from the bottom to the summit. After finishing the pyrolysis process (two days), RhB was watered to achieve ambient temperature.</p>
                <p>MB was produced by a poor-oxygen pyrolysis process under a traditional bell-shaped charcoal production kiln for a 30-day batch. The kiln was made from baked bricks, clay, and sand mortar. The kiln&#x2019;s structure comprises a bell-shaped heating firewood chamber, a door used for firewood loading, and biochar unloading. A combustion chamber provided hot air for the carbonization process, while four chimneys were installed around the heating chamber discharging smoke during the carbonization process. Firewood was fully loaded according to each layer underneath the heating chamber; the lowest layer was kept 10 cm away from the ground to ensure air convection. Before starting, the door was closed to begin the carbonization process. Air heating from the combustion chamber was slowly provided to the inner heating chamber to form carbonization. After 30 days of pyrolysis, the heating was switched off, and the combustion chamber was blocked off for an additional 15 days to cool to ambient temperature. The images of RhB and MB and their properties are shown in 
                    <xref ref-type="fig" rid="f1">Figure 1</xref> and 
                    <xref ref-type="table" rid="T1">Table 1</xref>, respectively.</p>
                <fig fig-type="figure" id="f1" orientation="portrait" position="float">
                    <label>Figure 1. </label>
                    <caption>
                        <title>Scanning electron microscope (SEM) images of biochar produced from rice husk (a) and melaleuca (b) at X800 magnification.</title>
                    </caption>
                    <graphic id="gr1" orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/77750/7bb49ef0-b454-4111-85c7-a86f61bd24f7_figure1.gif"/>
                </fig>
                <table-wrap id="T1" orientation="portrait" position="float">
                    <label>Table 1. </label>
                    <caption>
                        <title>Main properties of biochar derived from rice husk and melaleuca used in the field experiment.</title>
                    </caption>
                    <table content-type="article-table" frame="hsides">
                        <thead>
                            <tr>
                                <th align="left" colspan="1" rowspan="1" valign="top">Items</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Rice husk</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Melaleuca</th>
                            </tr>
                        </thead>
                        <tbody>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">pH (H
                                    <sub>2</sub>O)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">9.56</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">7.54</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">EC (mS cm
                                    <sup>&#x2212;1</sup>)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.78</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.28</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">CEC (cmol (+) kg
                                    <sup>&#x2212;1</sup>)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">13.2</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">9.55</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Total C (g kg
                                    <sup>&#x2212;1</sup>)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">253.5</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">291.8</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Total N (g kg
                                    <sup>&#x2212;1</sup>)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">3.26</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2.50</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Total P (g kg
                                    <sup>&#x2212;1</sup>)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.13</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.33</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Specific surface area (m
                                    <sup>2</sup> g
                                    <sup>&#x2212;1</sup>)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">51.93</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2.04</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Total pore volume (cm
                                    <sup>3</sup> g
                                    <sup>&#x2212;1</sup>)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.026</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.001</td>
                            </tr>
                        </tbody>
                    </table>
                </table-wrap>
            </sec>
            <sec id="sec5">
                <title>Experimental design</title>
                <p>The size of each experimental plot was 20 m
                    <sup>2</sup> (4 m &#x00d7; 5 m) which were arranged in a randomized complete block design with three replications. Each plot was separated by soil banks and covered with mulch film. Five treatments with RhB and MB incorporated into the soil paddy field comprised 0 t ha
                    <sup>&#x2212;1</sup> (conventional rice cultivation without biochar supplementation), 5 t ha
                    <sup>&#x2212;1</sup>, and 10 t ha
                    <sup>&#x2212;1</sup> (based dried weight) named CT0, RhB5, RhB10, MB5, and MB10, respectively. Biochar was manually spread on the soil surface of each pot and evenly incorporated into the plow layer of soil (approximately 20 cm) by shovels and rakes before sowing. Biochar additions were applied one time solely at the beginning of the experiment.</p>
            </sec>
            <sec id="sec6">
                <title>Rice cultivation and water management</title>
                <p>According to the local crop calendar, the experiment time corresponded with the Spring-Summer (SS) season (the second crop) (
                    <xref ref-type="table" rid="T2">Table 2</xref>). This is a transitional season between the dry and wet seasons. Rice straw and rice stubble from the previous rice crop cycle (Winter-Spring) were plowed by a hand tractor and underwent a 7-day fallow period before sowing. A short-duration variety of rice (IR50404 cultivar) typically grown in VMD was used in this field experiment (85-90 days of maturity). Pre-germinated seeds were sown on the wet-levelled soil using drum seeders at a rate equivalent to 120 kg ha
                    <sup>&#x2212;1</sup>. The irrigation followed regionally typical water management based on the farmer&#x2019;s practical experience.</p>
                <table-wrap id="T2" orientation="portrait" position="float">
                    <label>Table 2. </label>
                    <caption>
                        <title>Rice cropping calendar in the field experiment.</title>
                    </caption>
                    <table content-type="article-table" frame="hsides">
                        <thead>
                            <tr>
                                <th align="left" colspan="1" rowspan="1" valign="top">Cultivated schedule</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Date of experiment
                                    <xref ref-type="table-fn" rid="tfn1">
                                        <sup>1)</sup>
                                    </xref>
                                </th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Days after sowing</th>
                            </tr>
                        </thead>
                        <tbody>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Plowing</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">14/03/2019</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">&#x2212;7</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Biochar incorporation</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">21/03/2019</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Sowing</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">21/03/2021</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Starting irrigation</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">29/03/2021</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">8</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Fertilization</td>
                                <td colspan="1" rowspan="1"/>
                                <td colspan="1" rowspan="1"/>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">1
                                    <sup>st</sup> topdressing (16-8-20)
                                    <xref ref-type="table-fn" rid="tfn2">
                                        <sup>2)</sup>
                                    </xref>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">30/03/2019</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">9</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">2
                                    <sup>nd</sup> topdressing (32-16-0)
                                    <xref ref-type="table-fn" rid="tfn2">
                                        <sup>2)</sup>
                                    </xref>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">13/04/2019</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">23</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">3
                                    <sup>rd</sup> topdressing (32-16-20)
                                    <xref ref-type="table-fn" rid="tfn2">
                                        <sup>2)</sup>
                                    </xref>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">27/04/2019</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">38</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Drainage</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">30/05/2019</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">70</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Harvest</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">14/06/2019</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">85</td>
                            </tr>
                        </tbody>
                    </table>
                    <table-wrap-foot>
                        <fn-group content-type="footnotes">
                            <fn id="tfn1">
                                <label>
                                    <sup>1)</sup>
                                </label>
                                <p>dd/mm/yyyy.</p>
                            </fn>
                            <fn id="tfn2">
                                <label>
                                    <sup>2)</sup>
                                </label>
                                <p>The numbers in parenthesis indicate the amount (kg ha
                                    <sup>&#x2212;1</sup>) of fertilizers applied in terms of N, P and K, respectively.</p>
                            </fn>
                        </fn-group>
                    </table-wrap-foot>
                </table-wrap>
            </sec>
            <sec id="sec7">
                <title>Fertilizer application</title>
                <p>Inorganic fertilizers with the total amount of 80 kg N ha
                    <sup>&#x2212;1</sup>, 40 kg P
                    <sub>2</sub>O
                    <sub>5</sub> ha
                    <sup>&#x2212;1</sup> and 40 kg KCl ha
                    <sup>&#x2212;1</sup> were applied. The fertilization was divided into intervals at 9, 23, and 38 days after sowing (DAS) by broadcasting. Nitrogen (N) was applied as urea at a rate of 16-32-32 kg N ha
                    <sup>&#x2212;1</sup> (broadcasted three times). Phosphorus (P) was applied as superphosphate at a rate of 8-16-16 kg P
                    <sub>2</sub>O
                    <sub>5</sub> ha
                    <sup>&#x2212;1</sup> tolerant (broadcasted three times). Whereas potassium (K) was applied as potassium chloride at a 20-0-20 kg KCl ha
                    <sup>&#x2212;1</sup> rate (broadcasted twice). The rice cropping calendar and fertilizer application are shown in 
                    <xref ref-type="table" rid="T2">Table 2</xref>.</p>
            </sec>
            <sec id="sec8">
                <title>Measurements</title>
                <p>Scanning electron microscope (SEM) images of RhB and MB were captured by microscope (TM-1000, Hitachi, Japan). Specific surface area and total pore volume were determined using BET Surface Area Analyzer (Quatachrome Nova 1000e, USA).</p>
                <p>A weather station (WS-GP1, Delta-T Devices, Cambridge, UK) was installed on-site to record hourly temperature and rainfall at the experimental site. Redox potential (Eh) at plow-layer soil (20 cm) was measured by using platinum-tipped electrodes pined into the ground at a depth of 5, 10, and 20 cm; a portable Eh meter (HM31P; TOA-DKK, Japan) was connected to the electrodes to record soil Eh values at corresponding times to gas sampling. Surface water levels were also recorded simultaneously with gas sampling, using a ruler to read values directly in a plastic-perforated tube pre-installed in each plot.</p>
                <p>Topsoil samples (0-20 cm) in each plot were collected before adding biochar and harvest by an auger 3 cm diameter. Visible remaining biomass was eliminated before air drying and sieved at 2.0 mm. Initial soil samples (
                    <italic toggle="yes">n =</italic> 15) were mixed into a collective sample for analysis. Harvest soil samples were collected for each plot separately. Physical soil properties were measured as follows: soil texture - Pipette Robinson method (
                    <xref ref-type="bibr" rid="ref6">Carter and Gregorich, 2008</xref>), bulk density - Core method, and the particle density of soil (
                    <xref ref-type="bibr" rid="ref2">Blake and Hartge, 1986</xref>). Biochar and soil chemical properties were detected as follows: pH (H
                    <sub>2</sub>O) &#x2013; a portable pH meter (HANA, Germany), soil organic matter (SOM) and total organic C (TOC) &#x2013; 
                    <xref ref-type="bibr" rid="ref55">Walkley and Black (1934)</xref>, total P - 
                    <xref ref-type="bibr" rid="ref3">Bowman (1988)</xref>, available P (AP) - 
                    <xref ref-type="bibr" rid="ref38">Olsen and Sommers (1982)</xref>, total N &#x2013; semi-micro Kjeldahl method (
                    <xref ref-type="bibr" rid="ref4">Bremner, 1996</xref>), anaerobically mineralized N (AN)&#x2013; a 7-day anaerobic incubation at 40 &#x00b0;C (
                    <xref ref-type="bibr" rid="ref22">Keeney and Bremner, 1996</xref>), and CEC and exchangeable cations &#x2013; 
                    <xref ref-type="bibr" rid="ref49">Thomas (1982)</xref>.</p>
                <p>Rice yield was determined by harvesting from a 2.5 m &#x00d7; 2.0 m area in each plot at physiological maturity and removed unfilled grains by water before sun drying. A grain moisture tester (Riceter f2, Kett Electric Laboratory, Tokyo, Japan) was used to measure moisture content. The presented rice yield was adjusted to a 14% moisture content.</p>
                <p>The closed chamber method was used to collect gas samples. A chamber was made from transparent polyvinyl chloride (PVC) panels with a 1.5 mm thickness. The cross-sectional area was 0.25 m
                    <sup>2</sup> (0.5 m &#x00d7; 0.5 m). The height of the chamber was 70 cm from the bottom to the top layer. The chamber inside was equipped with a circulating fan, a temperature meter, and a pressure control plastic bag as described in detail by 
                    <xref ref-type="bibr" rid="ref30">Minamikawa (2015)</xref>. The chamber was placed on a plastic pre-installed base (a groove 4.5 cm depth) in each plot and sealing off by water before sampling. After chamber closure, a syringe (50 mL) was utilized to take inside gas at 1, 11, 21, and 31 minutes. Then, gas samples were injected into a 20-mL evacuated vial. The gas sampling was carried out from 10 DAS to 73 DAS at 7-day intervals. The concentrations of CH
                    <sub>4</sub> and N
                    <sub>2</sub>O were analyzed with a gas chromatograph (8610C, SRI Instruments, CA, USA) equipped with a flame ionization detector (FID) and an electron capture detector (ECD) for the analysis of CH
                    <sub>4</sub> and N
                    <sub>2</sub>O, respectively. The columns for the analysis of CH
                    <sub>4</sub> and N
                    <sub>2</sub>O were packed with Porapak Q (50&#x2013;80 mesh); dinitrogen (N
                    <sub>2</sub>) was used as the carrier gas for both FID and ECD.</p>
                <p>Porosity was calculated by dividing volume pores (based on the subtraction between bulk density and particle density of soils) by volume total (
                    <xref ref-type="bibr" rid="ref15">Flint and Flint, 2002</xref>). CH
                    <sub>4</sub> and N
                    <sub>2</sub>O fluxes were calculated by a linear progression of gas concentration change over time, and total fluxes of CH
                    <sub>4</sub> and N
                    <sub>2</sub>O were calculated using a trapezoidal integration method described by 
                    <xref ref-type="bibr" rid="ref30">Minamikawa (2015)</xref>. Global warming potential (GWP) was calculated based on CO
                    <sub>2</sub> equivalence (1 CH
                    <sub>4</sub> = 34 CO
                    <sub>2</sub>-eq; 1 N
                    <sub>2</sub>O = 298 CO
                    <sub>2</sub>-eq) at a 100-year scale of climate-carbon feedbacks (
                    <xref ref-type="bibr" rid="ref31">Myhre 
                        <italic toggle="yes">et al.</italic>, 2013</xref>). Yield-scale GWP was calculated by dividing the GWP by grain yield (
                    <xref ref-type="bibr" rid="ref29">Minamikawa 
                        <italic toggle="yes">et al.</italic>, 2021</xref>).</p>
            </sec>
            <sec id="sec9">
                <title>Statistical analysis</title>
                <p>One-Way analysis of variance (ANOVA) was used to assess the effects of each biochar on grain yield, gas fluxes, GWP, yield-scale GWP, and soil improvement. The difference of treatments was carried out using Duncan&#x2019;s method for all pairwise multiple comparison procedures. Linear regression analyses were performed to assess the relationship between Eh change and methane emission. We also analyzed the relationship between biochar application rate and gas emissions. In the statistical analysis, we did not compare the difference between RhB and MB. All analyses were carried out using 
                    <ext-link ext-link-type="uri" xlink:href="http://www.r-project.org/">R</ext-link> 
                    <ext-link ext-link-type="uri" xlink:href="https://stat.ethz.ch/R-manual/R-devel/library/stats/html/00Index.html">stats</ext-link> Version 4.2.0 (R Project for Statistical Computing, RRID:SCR_001905). The results are presented in tabular form with the values including mean &#x00b1; standard deviation (SD) and the different symbols with a confidence level of 95%.</p>
            </sec>
        </sec>
        <sec id="sec10" sec-type="results">
            <title>Results</title>
            <sec id="sec11">
                <title>Weather and water management</title>
                <p>The mean air temperature and the total rainfall during the experiment were 28.9 &#x00b0;C and 429 mm, respectively (
                    <xref ref-type="fig" rid="f2">Figure 2</xref>). High rainfall was observed between &#x2013;40-60 and &#x2013;65-80 DAS. 
                    <xref ref-type="fig" rid="f3">Figure 3</xref> shows that the flooding water regime was predominantly observed during the experimental regression. The trend of water levels variation was similar over treatments. Water was irrigated from 7 DAS, reflooded 3-5 cm from soil surface for fertilizing (9, 23, and 38 DAS) and respective multiple drainage practice (&#x2212;10 to 5 cm) (
                    <xref ref-type="bibr" rid="ref52">Uno 
                        <italic toggle="yes">et al</italic>., 2021</xref>) was carried out for the remaining periods. Fifteen days before harvesting (70 DAS), the soil was drained and kept saturated to minimize rice lodging and easy-to-harvest grain. Rice plants flowered and headed during 45-60 DAS.</p>
                <fig fig-type="figure" id="f2" orientation="portrait" position="float">
                    <label>Figure 2. </label>
                    <caption>
                        <title>Temperature and rainfall during the field experiment.</title>
                    </caption>
                    <graphic id="gr2" orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/77750/7bb49ef0-b454-4111-85c7-a86f61bd24f7_figure2.gif"/>
                </fig>
                <fig fig-type="figure" id="f3" orientation="portrait" position="float">
                    <label>Figure 3. </label>
                    <caption>
                        <title>Time course changes in soil redox potential (Eh), water level, hourly CH
                            <sub>4</sub> and N
                            <sub>2</sub>O fluxes in the paddy field applied without (left) or with RhB (center) or MB (right) during the field experiment.</title>
                        <p>Error bars indicate the standard error (
                            <italic toggle="yes">n</italic> = 3). Vertical dotted lines illustrate agronomic management of the first, the second and the third topdressing fertilizer (F
                            <sub>1</sub>, F
                            <sub>2</sub> and F
                            <sub>3</sub>, respectively), drainage (D) and harvest (H).</p>
                    </caption>
                    <graphic id="gr3" orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/77750/7bb49ef0-b454-4111-85c7-a86f61bd24f7_figure3.gif"/>
                </fig>
            </sec>
            <sec id="sec12">
                <title>CH
                    <sub>4</sub> and N
                    <sub>2</sub>O emissions</title>
                <p>CH
                    <sub>4</sub> emissions gradually increased in the early rice growth stage (0-17 DAS) and almost stopped after drainage (70 DAS) (
                    <xref ref-type="fig" rid="f3">Figure 3</xref>). It should be noted that CH
                    <sub>4</sub> flux was predominant in the period from 17 &#x2013; 59 DAS and several CH
                    <sub>4</sub> flux peaks were observed between treatments (i.e., three peaks were observed in MB5 and MB10). Maximum CH
                    <sub>4</sub> flux peaks reached simultaneously in all treatments after 31 DAS. Highest peaks between treatments are represented in a descending way as follows: CT0 &gt; MB5 &gt; MB10 &gt; RhB5 &gt; RhB10. Compared to the CT0 treatment, biochar application reduced total CH
                    <sub>4</sub> emissions significantly (
                    <xref ref-type="table" rid="T3">Table 3</xref>). Particularly, RhB5 and RhB10 mitigated total CH
                    <sub>4</sub> flux from 24.2 to 28.0%, respectively, while MB5 and MB10 alleviated between 22.0 and 14.1%, respectively. Irrespective of RhB and MB, the CH
                    <sub>4</sub> flux was insignificant with an increasing biochar addition rate from 5 to 10 t ha
                    <sup>&#x2212;1</sup> (
                    <italic toggle="yes">P</italic> &lt; 0.01). There was a negative linear regression relationship between biochar application rate and total CH
                    <sub>4</sub> emission (
                    <italic toggle="yes">P &lt;</italic> 0.001, 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.825) (
                    <xref ref-type="fig" rid="f4">Figure 4</xref>). In contrast, the linear regression of melaleuca biochar was poorly explained with increasing biochar amendment rate and total CH
                    <sub>4</sub> flux (
                    <italic toggle="yes">P =</italic> 0.095, 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.254).</p>
                <table-wrap id="T3" orientation="portrait" position="float">
                    <label>Table 3. </label>
                    <caption>
                        <title>Grain, total CH
                            <sub>4</sub> and N
                            <sub>2</sub>O fluxes, global warming potential (GWP) and yield-scaled GWP at 100 years scale in the paddy field applied without or with biochar
                            <xref ref-type="table-fn" rid="tfn3">
                                <sup>1)</sup>
                            </xref>.</title>
                    </caption>
                    <table content-type="article-table" frame="hsides">
                        <thead>
                            <tr>
                                <th align="left" colspan="1" rowspan="1" valign="top">Treatment
                                    <xref ref-type="table-fn" rid="tfn4">
                                        <sup>2)</sup>
                                    </xref>
                                </th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Grain (g m
                                    <sup>&#x2212;2</sup>)</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">CH
                                    <sub>4</sub> (g CH
                                    <sub>4</sub> m
                                    <sup>&#x2212;2</sup>)</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">N
                                    <sub>2</sub>O (g N
                                    <sub>2</sub>O m
                                    <sup>&#x2212;2</sup>)</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">GWP (g CO
                                    <sub>2</sub>-eq m
                                    <sup>&#x2212;2</sup>)</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Yield-scaled GWP (g CO
                                    <sub>2</sub>-eq m
                                    <sup>&#x2212;2</sup>)</th>
                            </tr>
                        </thead>
                        <tbody>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">CT0</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">498 &#x00b1; 47.6</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">18.6 &#x00b1; 0.80
                                    <sup>aA</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.39 &#x00b1; 0.07
                                    <sup>aA</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">749 &#x00b1; 13.5
                                    <sup>aA</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">1.51 &#x00b1; 0.13
                                    <sup>aA</sup>
                                </td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">RhB5</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">513 &#x00b1; 56.2</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">14.1 &#x00b1; 0.23
                                    <sup>b</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.29 &#x00b1; 0.07
                                    <sup>ab</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">566 &#x00b1; 25.5
                                    <sup>b</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">1.12 &#x00b1; 0.18
                                    <sup>b</sup>
                                </td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">RhB10</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">510 &#x00b1; 33.0</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">13.4 &#x00b1; 0.30
                                    <sup>b</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.23 &#x00b1; 0.04
                                    <sup>b</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">524 &#x00b1; 2.76
                                    <sup>c</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">1.03 &#x00b1; 0.06
                                    <sup>c</sup>
                                </td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">MB5</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">519 &#x00b1; 9.86</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">14.5 &#x00b1; 1.00
                                    <sup>B</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.24 &#x00b1; 0.01
                                    <sup>B</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">563 &#x00b1; 31.6
                                    <sup>B</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">1.09 &#x00b1; 0.08
                                    <sup>B</sup>
                                </td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">MB10</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">517 &#x00b1; 10.9</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">15.9 &#x00b1; 0.90
                                    <sup>B</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.17 &#x00b1; 0.07
                                    <sup>B</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">591 &#x00b1; 10.8
                                    <sup>B</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">1.14 &#x00b1; 0.44
                                    <sup>B</sup>
                                </td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">
                                    <italic toggle="yes">P</italic> value
                                    <xref ref-type="table-fn" rid="tfn5">
                                        <sup>3)</sup>
                                    </xref>
                                </td>
                                <td colspan="1" rowspan="1"/>
                                <td colspan="1" rowspan="1"/>
                                <td colspan="1" rowspan="1"/>
                                <td colspan="1" rowspan="1"/>
                                <td colspan="1" rowspan="1"/>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">CT &#x00d7; RhB</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">&#x2020;</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">***</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">*</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">***</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">**</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">CT &#x00d7; MB</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">&#x2020;</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">**</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">**</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">***</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">**</td>
                            </tr>
                        </tbody>
                    </table>
                    <table-wrap-foot>
                        <fn-group content-type="footnotes">
                            <fn id="tfn3">
                                <label>
                                    <sup>1)</sup>
                                </label>
                                <p>Data represent as means &#x00b1; SD (
                                    <italic toggle="yes">n</italic> = 3).</p>
                            </fn>
                            <fn id="tfn4">
                                <label>
                                    <sup>2)</sup>
                                </label>
                                <p>CT0, control treatment; RhB5 and RhB10, 5 and 10 t ha
                                    <sup>&#x2212;1</sup> rice-husk biochar amendment, respectively; MB5 and MB10, 5 and 10 t ha
                                    <sup>&#x2212;1</sup> melaleuca biochar amendment, respectively.</p>
                            </fn>
                            <fn id="tfn5">
                                <label>
                                    <sup>3)</sup>
                                </label>
                                <p>Statistical analysis did not compare between RhB and MB. The letters indicate significant difference according to Duncan&#x2019;s multiple range test (***
                                    <italic toggle="yes">P</italic> &lt; 0.001, **
                                    <italic toggle="yes">P</italic> &lt; 0.01, *
                                    <italic toggle="yes">P</italic> &lt; 0.05 and &#x2020;
                                    <italic toggle="yes">P</italic> &gt; 0.05). Normal and capital lowercases indicate a significant difference between CT0 vs. RhB and CT0 vs. MB, respectively.</p>
                            </fn>
                        </fn-group>
                    </table-wrap-foot>
                </table-wrap>
                <fig fig-type="figure" id="f4" orientation="portrait" position="float">
                    <label>Figure 4. </label>
                    <caption>
                        <title>Relationship between biochar application rate and total CH
                            <sub>4</sub> (above) and N
                            <sub>2</sub>O (below) fluxes during the field experiment.</title>
                        <p>Each symbol represents one replication in each treatment.</p>
                    </caption>
                    <graphic id="gr4" orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/77750/7bb49ef0-b454-4111-85c7-a86f61bd24f7_figure4.gif"/>
                </fig>
                <p>N
                    <sub>2</sub>O was released mainly in the early stage of rice growth in all treatments (
                    <xref ref-type="fig" rid="f3">Figure 3</xref>). The highest N
                    <sub>2</sub>O flux peaks were observed in the CT0 (24 DAS). All measured values were below 1.5 mg N
                    <sub>2</sub>O m
                    <sup>&#x2212;2</sup> h
                    <sup>&#x2212;1</sup>. As observed, N
                    <sub>2</sub>O flux flushed mainly during the fertilizing period from 9 to 38 DAS, even though experimental pots were predominantly flooded, especially in the CT0 accounted for 56.8% in total, while RhB and MB varied by 50.6-53.1% and 52.3-47.6%, respectively. Total N
                    <sub>2</sub>O emission was reduced in RhB or MB applied soil compared to CT0 (
                    <xref ref-type="table" rid="T3">Table 3</xref>). Specifically, RhB10 significantly reduced by approximately 41.0%, whereas MB5 and MB10 by 38.5 and 56.4%, respectively. However, the reduction of total N
                    <sub>2</sub>O flux was insignificant in MB5. As a result, there were different negative linear relationships of biochar application rate and total N
                    <sub>2</sub>O flux (RhB, 
                    <italic toggle="yes">P =</italic> 0.012, 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.619; MB, 
                    <italic toggle="yes">P =</italic> 0.002, 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.757) (
                    <xref ref-type="fig" rid="f4">Figure 4</xref>).</p>
            </sec>
            <sec id="sec13">
                <title>Rice yield, GWP, and yield-scaled GWP</title>
                <p>Biochar addition to the soil slightly increased rice yield compared to the CT0, but the statistical analysis was insignificant (
                    <xref ref-type="table" rid="T3">Table 3</xref>). A similar pattern about emissions was seen among GWP, yield-scaled GWP, and total CH
                    <sub>4</sub> flux due to CH
                    <sub>4</sub> flux was greatest contribute to GWP, yield-scaled GWP. The RhB additions significantly decreased the GWP and yield-scaled GWP by 24.4 &#x2013; 30.0% and 25.8 &#x2013; 31.8% for RhB5 and RhB10, respectively. Although MB significantly diminished the GWP and yield-scaled GWP by 24.8 &#x2013; 21.09% and 27.8 &#x2013; 24.5%, respectively, there was no significant difference between MB5 and MB10.</p>
            </sec>
            <sec id="sec14">
                <title>Soil characteristics</title>
                <p>A similar performance pattern of soil Eh condition was seen among treatments (
                    <xref ref-type="fig" rid="f3">Figure 3</xref>). Eh reduced after initial irrigation and was seen to reach a stable level (below -250 mV) during the rice growth period from 17 to 66 DAS. Whereas the final drainage rapidly increased the soil Eh condition (73 DAS) in all treatments. The supplementation of RhB and MB obviously improved soil Eh condition compared to the CT0 by 7.44 &#x2013; 14.5% and 10.7 &#x2013; 19.0%, respectively (
                    <xref ref-type="table" rid="T4">Table 4</xref>). There was a negative linear relationship between hourly CH
                    <sub>4</sub> flux and the Eh values in RhB (
                    <italic toggle="yes">P &lt;</italic> 0.001; 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.552) and MB (
                    <italic toggle="yes">P &lt;</italic> 0.001; 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.502) (
                    <xref ref-type="fig" rid="f5">Figure 5</xref>).</p>
                <table-wrap id="T4" orientation="portrait" position="float">
                    <label>Table 4. </label>
                    <caption>
                        <title>Physiochemical properties
                            <xref ref-type="table-fn" rid="tfn6">
                                <sup>1)</sup>
                            </xref> of soil applied without or with biochar.</title>
                    </caption>
                    <table content-type="article-table" frame="hsides">
                        <thead>
                            <tr>
                                <th align="left" colspan="1" rowspan="1" valign="top">Treatment
                                    <xref ref-type="table-fn" rid="tfn7">
                                        <sup>2)</sup>
                                    </xref>
                                </th>
                                <th align="left" colspan="1" rowspan="1" valign="top">pH</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Eh
                                    <xref ref-type="table-fn" rid="tfn7">
                                        <sup>3)</sup>
                                    </xref> (mV)</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Bulk density (g cm
                                    <sup>&#x2212;3</sup>)</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Porosity (%)</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">SOM (g kg
                                    <sup>&#x2212;1</sup>)</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">AP (mg kg
                                    <sup>&#x2212;1</sup>)</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">AN (mg kg
                                    <sup>&#x2212;1</sup>)</th>
                            </tr>
                        </thead>
                        <tbody>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">CT0</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">4.69 &#x00b1; 0.10</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">-242 &#x00b1; 12.3
                                    <sup>cB</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.97 &#x00b1; 0.10
                                    <sup>aA</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">53.3 &#x00b1; 0.75
                                    <sup>bB</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">31.8 &#x00b1; 0.36
                                    <sup>cC</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">19.0 &#x00b1; 3.95
                                    <sup>B</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">10.7 &#x00b1; 1.03
                                    <sup>bB</sup>
                                </td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">RhB5</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">4.81 &#x00b1; 0.21</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">-224 &#x00b1; 6.12
                                    <sup>b</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.78 &#x00b1; 0.04
                                    <sup>b</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">61.5 &#x00b1; 7.20
                                    <sup>ab</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">43.2 &#x00b1; 2.08
                                    <sup>b</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">21.8 &#x00b1; 3.48</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">15.5 &#x00b1; 0.52
                                    <sup>a</sup>
                                </td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">RhB10</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">5.26 &#x00b1; 0.64</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">-207 &#x00b1; 2.13
                                    <sup>a</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.74 &#x00b1; 0.13
                                    <sup>b</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">65.1 &#x00b1; 2.07
                                    <sup>a</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">47.6 &#x00b1; 1.13
                                    <sup>a</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">25.2 &#x00b1; 4.04</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">14.8 &#x00b1; 0.60
                                    <sup>a</sup>
                                </td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">MB5</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">4.82 &#x00b1; 0.13</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">-216 &#x00b1; 16.3
                                    <sup>A</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.75 &#x00b1; 0.06
                                    <sup>B</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">55.5 &#x00b1; 1.92
                                    <sup>A</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">39.1 &#x00b1; 3.03
                                    <sup>B</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">25.1 &#x00b1; 2.81
                                    <sup>AB</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">14.5 &#x00b1; 1.10
                                    <sup>AB</sup>
                                </td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">MB10</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">4.68 &#x00b1; 0.17</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">-196 &#x00b1; 5.78
                                    <sup>A</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.71 &#x00b1; 0.05
                                    <sup>B</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">62.9 &#x00b1; 4.19
                                    <sup>A</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">45.0 &#x00b1; 1.30
                                    <sup>A</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">28.8 &#x00b1; 2.79
                                    <sup>A</sup>
                                </td>
                                <td align="left" colspan="1" rowspan="1" valign="top">16.6 &#x00b1; 2.97
                                    <sup>A</sup>
                                </td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">
                                    <italic toggle="yes">P</italic> value
                                    <xref ref-type="table-fn" rid="tfn8">
                                        <sup>4)</sup>
                                    </xref>
                                </td>
                                <td colspan="1" rowspan="1"/>
                                <td colspan="1" rowspan="1"/>
                                <td colspan="1" rowspan="1"/>
                                <td colspan="1" rowspan="1"/>
                                <td colspan="1" rowspan="1"/>
                                <td colspan="1" rowspan="1"/>
                                <td colspan="1" rowspan="1"/>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">CT &#x00d7; RhB</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">&#x2020;</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">*</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">*</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">*</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">***</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">&#x2020;</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">***</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">CT &#x00d7; MB</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">&#x2020;</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">*</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">***</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">*</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">***</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">*</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">*</td>
                            </tr>
                        </tbody>
                    </table>
                    <table-wrap-foot>
                        <fn-group content-type="footnotes">
                            <fn id="tfn6">
                                <label>
                                    <sup>1)</sup>
                                </label>
                                <p>Data represent as means &#x00b1; SD (
                                    <italic toggle="yes">n</italic> = 3).</p>
                            </fn>
                            <fn id="tfn7">
                                <label>
                                    <sup>2)</sup>
                                </label>
                                <p>Abbreviations are the same as 
                                    <xref ref-type="table" rid="T3">Table 3</xref>.</p>
                            </fn>
                            <fn id="tfn8">
                                <label>
                                    <sup>3)</sup>
                                </label>
                                <p>Mean value is based on the whole values measured during the experimentation in each plot at 3 soil levels depth comprising 5, 10 and 20 cm between 10 and 64 DAS.</p>
                            </fn>
                            <fn id="tfn9">
                                <label>
                                    <sup>4)</sup>
                                </label>
                                <p>Statistical analysis was carried out as the same as 
                                    <xref ref-type="table" rid="T3">Table 3</xref>.</p>
                            </fn>
                        </fn-group>
                    </table-wrap-foot>
                </table-wrap>
                <fig fig-type="figure" id="f5" orientation="portrait" position="float">
                    <label>Figure 5. </label>
                    <caption>
                        <title>Relationship between the hourly CH
                            <sub>4</sub> flux and Eh in the field applied with RhB (above) or MB (below).</title>
                    </caption>
                    <graphic id="gr5" orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/77750/7bb49ef0-b454-4111-85c7-a86f61bd24f7_figure5.gif"/>
                </fig>
                <p>
                    <xref ref-type="table" rid="T4">Table 4</xref> represents the soil characteristic differences between treatments at the time of harvest. Overall, although biochar amendment was seen to increase soil pH slightly, statistical analysis implied no significant difference between treatments. Yet, biochar amendment significantly reduced the soil bulk density (RhB5, 19%; RhB10, 23%; MB5, 22.7% and MB10 26.8%) and ameliorated the soil porosity (RhB5, 8.2%; RhB10, 11.8%; MB5, 2.2%, and MB10 9.6%). However, increasing RhB and MB biochar application rate from 5 to 10 t ha
                    <sup>&#x2212;1</sup> did not significantly change soil bulk density and porosity. Moreover, intensifying biochar incorporation significantly increased SOM by 38.6 &#x2013; 52.7% for RhB and 25.4 &#x2013; 45.9% for MB. Notably, AN in biochar-applied treatments was higher than that of the CT0 by 44.8 &#x2013; 38.3% and 35.5 &#x2013; 55.1% for RhB and MB, respectively. AP significantly increased in the MB treatments by 32.1 &#x2013; 51.58% but did not in RhB. Although additional biochar increased the available and mineralized nutrients, statistical analysis results showed no significant difference between biochar application rates of 5 to 10 t ha
                    <sup>&#x2212;1</sup> (
                    <xref ref-type="table" rid="T4">Table 4</xref>) (
                    <xref ref-type="bibr" rid="ref51">Tran Sy 
                        <italic toggle="yes">et al</italic>., 2021</xref>).</p>
            </sec>
        </sec>
        <sec id="sec15" sec-type="discussion">
            <title>Discussion</title>
            <sec id="sec16">
                <title>Effects of biochar incorporation on CH
                    <sub>4</sub> and N
                    <sub>2</sub>O fluxes</title>
                <p>Conventional practices without biochar application released 18.6 g CH
                    <sub>4</sub> m
                    <sup>&#x2212;2</sup> and 0.39 g N
                    <sub>2</sub>O m
                    <sup>&#x2212;2</sup> (
                    <xref ref-type="table" rid="T3">Table 3</xref>). These values are in accordance with previous findings conducted in the VMD (
                    <xref ref-type="bibr" rid="ref54">Vo 
                        <italic toggle="yes">et al.</italic> 2020</xref>; 
                    <xref ref-type="bibr" rid="ref29">Minamikawa 
                        <italic toggle="yes">et al.</italic> 2021</xref>; 
                    <xref ref-type="bibr" rid="ref52">Uno 
                        <italic toggle="yes">et al.</italic> 2021</xref>). Notably, RhB and MB amendments under typically local water management, and conventional practices significantly reduced CH
                    <sub>4</sub> flux by 24.2 % in RhB5, 28.0 % in RhB10, 22.0 % in MB5, 14.1 % in MB10 and N
                    <sub>2</sub>O flux by 38.5 % in RhB5, 56.4 % in RhB10, 25.6 % in MB5, 41.0 % in MB10, and slightly improved rice yield (2.41-4.21%) (
                    <xref ref-type="table" rid="T3">Table 3</xref>). Similarly, 
                    <xref ref-type="bibr" rid="ref68">Yang 
                        <italic toggle="yes">et al.</italic> (2019)</xref> demonstrated that biochar additions (20 - 40 t ha
                    <sup>&#x2212;1</sup>) under controlled irrigation in the Taihu Lake region, China mitigated both CH
                    <sub>4</sub> and N
                    <sub>2</sub>O emissions by 35.7% and 21.5%, respectively, and simultaneously enhanced rice yield by 16.7-24.3%. Moreover, 
                    <xref ref-type="bibr" rid="ref65">Wu 
                        <italic toggle="yes">et al.</italic> (2019b)</xref> reported that biochar additions (20 - 40 t ha
                    <sup>&#x2212;1</sup>) significantly decreased CH
                    <sub>4</sub> and N
                    <sub>2</sub>O fluxes by 11.2-17.5% and 19.5-26.3%, respectively, and increased grain yield by 7.9-9.2%. In line with our findings, a long-term biochar application (5 &#x2013; 10 t ha
                    <sup>&#x2212;1</sup>) in China's typical double rice plantation region also significantly decreased CH
                    <sub>4</sub> flux by 26.18% (
                    <xref ref-type="bibr" rid="ref41">Qin 
                        <italic toggle="yes">et al.</italic> 2016</xref>). Nevertheless, 
                    <xref ref-type="bibr" rid="ref57">Wang 
                        <italic toggle="yes">et al.</italic> (2011)</xref> reported that the biochar incorporation (50 ton ha
                    <sup>&#x2212;1</sup>) into the soil significantly decreased N
                    <sub>2</sub>O flux by 41.4-93.5% in lab-scale experiments. In parallel, a meta-analysis based on 30 studies with 261 experimental treatments (lab-scale and pilot-scale) from 2007 to 2013 demonstrated that the addition of biochar reduced N
                    <sub>2</sub>O emissions by 54% (
                    <xref ref-type="bibr" rid="ref8">Cayuela 
                        <italic toggle="yes">et al.</italic> 2014</xref>). However, 
                    <xref ref-type="bibr" rid="ref24">Koyama 
                        <italic toggle="yes">et al.</italic> (2015)</xref> reported that biochar application (10-40 t ha
                    <sup>&#x2212;1</sup>) reduced CH
                    <sub>4</sub> flux but did not N
                    <sub>2</sub>O. In the case of 
                    <xref ref-type="bibr" rid="ref26">Liu 
                        <italic toggle="yes">et al.</italic> (2014)</xref>, biochar supplementation (24-48 t ha
                    <sup>&#x2212;1</sup>) significantly reduced CH
                    <sub>4</sub> flux by 33.9-40.2%, while N
                    <sub>2</sub>O flux significantly increased by 150 to 190%. Overall, biochar amendment could reduce CH
                    <sub>4</sub> flux from a rice paddy field, but in some cases, the effect on N
                    <sub>2</sub>O flux remains uncertain. Our study demonstrated that rice husk and melaleuca biochar applications with a range of 5-10 t ha
                    <sup>&#x2212;1</sup> significantly reduced both CH
                    <sub>4</sub> and N
                    <sub>2</sub>O fluxes within a Thionic Glycesol soil in the VMD. Albeit, the biochar application rate between 5 and 10 t ha
                    <sup>&#x2212;1</sup> hardly obtained the disparity of CH
                    <sub>4</sub> and N
                    <sub>2</sub>O emissions. Thus, a wide range of biochar application amounts should be evaluated to provide more tailored recommendations.</p>
                <p>The CH
                    <sub>4</sub> mitigation by biochar application consistently pertains to the increasing soil oxidation rate and methanotrophs community. Although we did not determine the number of methanogens and methanotrophs, 
                    <xref ref-type="bibr" rid="ref34">Nan 
                        <italic toggle="yes">et al.</italic> (2021)</xref> demonstrated that biochar application stimulates the abundance in either methanogens or methanotrophs, with a high amount of methanotrophs detected in most cases resulted in decreasing of CH
                    <sub>4</sub> flux. Moreover, 
                    <xref ref-type="bibr" rid="ref64">Wu 
                        <italic toggle="yes">et al.</italic> (2019a)</xref> reported that biochar applications to fertilized paddy field soils increased the total type I 
                    <italic toggle="yes">pmo</italic>A (preferred the CH
                    <sub>4</sub> environment) and type II 
                    <italic toggle="yes">pmo</italic>A (more dynamic in low CH
                    <sub>4</sub> conditions) methanotrophs comparing to non-amended biochar, indicating that CH
                    <sub>4</sub> flux mitigation by promoting potential CH
                    <sub>4</sub> oxidation. Thus, we adopted a hypothesis that the balance of activities between methanogens and methanotrophs in a site-specific environment results in either an increasing or decreasing CH
                    <sub>4</sub> flux. 
                    <xref ref-type="bibr" rid="ref14">Feng 
                        <italic toggle="yes">et al.</italic> (2012)</xref> revealed the main mechanisms of CH
                    <sub>4</sub> flux reduction in a biochar-supplemented field were by (1) increased methanotrophic proteobacterial abundance significantly and (2) decreased the methanogenic to methanotrophic proportion substantially. Thus, an increase of methanotrophs dynamic in paddy field soil by biochar addition can be expected to play a vital role in mitigating CH
                    <sub>4</sub> fluxes. Our study demonstrated that rice husk and melaleuca biochar could promote low-GHG emissions in the rice production system in the VMD.</p>
                <p>We achieved N
                    <sub>2</sub>O flux reduction by incorporating biochar into the topsoil layer when compared to the non-amended biochar field. However, several hypotheses supposed that soil applied with biochar could not decrease the N
                    <sub>2</sub>O flux (
                    <xref ref-type="bibr" rid="ref24">Koyama 
                        <italic toggle="yes">et al.</italic>, 2015</xref>; 
                    <xref ref-type="bibr" rid="ref53">van Zwieten 
                        <italic toggle="yes">et al</italic>., 2010</xref>). Similar to our field study, several findings achieved a total N
                    <sub>2</sub>O flux reduction (
                    <xref ref-type="bibr" rid="ref42">Shaukat 
                        <italic toggle="yes">et al.</italic>, 2019</xref>; 
                    <xref ref-type="bibr" rid="ref69">Zhang 
                        <italic toggle="yes">et al</italic>., 2010</xref>). The mitigation of N
                    <sub>2</sub>O flux in biochar-treated soils could be attributed to soil moisture contents and nitrification processes (
                    <xref ref-type="bibr" rid="ref1">Ameloot 
                        <italic toggle="yes">et al.</italic>, 2016</xref>). In agreement with the hypothesis, 
                    <xref ref-type="bibr" rid="ref42">Shaukat 
                        <italic toggle="yes">et al.</italic> (2019)</xref> demonstrated that fields with biochar added retained 9-14% higher moisture contents than fields without biochar amended and resulted in a significant reduction of the N
                    <sub>2</sub>O flux. Supporting the idea, 
                    <xref ref-type="bibr" rid="ref56">Wang 
                        <italic toggle="yes">et al.</italic> (2013)</xref> revealed the relationship between the denitrifying community and N
                    <sub>2</sub>O flux change, where biochar supplementation significantly shifted the abundance of NO
                    <sub>3</sub>-utilizing bacteria (carrying the 
                    <italic toggle="yes">nir</italic>K and 
                    <italic toggle="yes">nir</italic>S genes), leading to less N
                    <sub>2</sub>O generation and more N
                    <sub>2</sub>O-consuming bacteria (carrying the 
                    <italic toggle="yes">nos</italic>Z gene). Moreover, 
                    <xref ref-type="bibr" rid="ref7">Cayuela 
                        <italic toggle="yes">et al.</italic> (2013)</xref> used 
                    <sup>15</sup>N gas-flux to observe the reduction of N
                    <sub>2</sub>O/(N
                    <sub>2</sub>+N
                    <sub>2</sub>O) and demonstrated that biochar facilitated the last step of denitrification. The key mechanisms of N
                    <sub>2</sub>O flux reduction under biochar amendment were by (i) stimulated nitrification generation via electron donation, a decrease in total denitrification by serving as an alternative electron acceptor by acting as electron shuttle to soil NO
                    <sub>3</sub>
                    <sup>&#x2212;</sup> consuming microorganisms (
                    <xref ref-type="bibr" rid="ref7">Cayuela 
                        <italic toggle="yes">et al.</italic>, 2013</xref>), and (ii) based on the entrapment of N
                    <sub>2</sub>O in water-saturated soil pores and co-occurrent stimulation of microbial N
                    <sub>2</sub>O reduction deriving in an overall decrease of the N
                    <sub>2</sub>O/(N
                    <sub>2</sub>O&#x2009;+&#x2009;N
                    <sub>2</sub>) ratio (
                    <xref ref-type="bibr" rid="ref19">Harter 
                        <italic toggle="yes">et al.</italic>, 2016</xref>). Therefore, biochar could be attributed as a decisive factor to inhibit N
                    <sub>2</sub>O production and simultaneously stimulate N
                    <sub>2</sub>O utilization. As such, these findings and the above-discussed mechanisms strongly support our findings in suggesting N2O flux reduction from biochar amendment in the rice paddy field.</p>
                <p>Our study showed that N
                    <sub>2</sub>O emission was mainly concentrated during fertilization, which indicates fertilization provides more available N driving for soil N
                    <sub>2</sub>O emission. 
                    <xref ref-type="bibr" rid="ref67">Xie 
                        <italic toggle="yes">et al.</italic> (2013)</xref> observed 
                    <sup>15</sup>N abundance significantly intensified by the application of 
                    <sup>15</sup>N-enriched urea. Our study did not measure NH
                    <sub>4</sub>
                    <sup>+</sup> or NO
                    <sub>3</sub>
                    <sup>&#x2212;</sup> concentration during fertilizing, so the mechanism remains uncertain. N
                    <sub>2</sub>O emission via the nitrification process directly pertains to soil physical, chemical, and biological properties (
                    <xref ref-type="bibr" rid="ref20">Huang 
                        <italic toggle="yes">et al.</italic>, 2019</xref>). Thus, we speculate that N fertilizing increased the nitrification activities and stimulated the strong metabolism of potential N
                    <sub>2</sub>O-producing bacteria. 
                    <xref ref-type="bibr" rid="ref29">Minamikawa 
                        <italic toggle="yes">et al.</italic> (2021)</xref> reported that higher N availability levels in soil than rice plant uptake demands resulted in increasing N
                    <sub>2</sub>O emissions. Although N-fertilizing obviously promoted N
                    <sub>2</sub>O emissions for the majority of time, N
                    <sub>2</sub>O emission peaks of biochar-amended soil were lower than that of biochar-unamended soil. This would indicate that biochar potentially changed the functionality and diversity of denitrifiers within the soils and inhibited the conversion of NO
                    <sub>2</sub>
                    <sup>&#x2212;</sup> and NO
                    <sub>3</sub>
                    <sup>&#x2212;</sup> to N
                    <sub>2</sub>O (
                    <xref ref-type="bibr" rid="ref69">Zhang 
                        <italic toggle="yes">et al.</italic>, 2010</xref>).</p>
                <p>Water management is a crucial factor in the strategy of GHGs reduction, although we achieved the GHGs reduction under typical water management when most of the time the soil was flooded. Multiple-flooded times in this study were due to the combination of high rainfall in the transition season (rainfall, 
                    <xref ref-type="fig" rid="f2">Figure 2</xref>; water level, 
                    <xref ref-type="fig" rid="f3">Figure 3</xref>) and the typical flooding water management practice of the farmers in the region. 
                    <xref ref-type="bibr" rid="ref52">Uno 
                        <italic toggle="yes">et al</italic>. (2021)</xref> conducted a 2-year field experiment in An Giang province in the VMD and demonstrated that AWD (known as multiple drainages) significantly reduced CH
                    <sub>4</sub> by 35%, while found no difference in N
                    <sub>2</sub>O emissions, but a 22% yield improved. Moreover, 
                    <xref ref-type="bibr" rid="ref29">Minamikawa 
                        <italic toggle="yes">et al.</italic> (2021)</xref> registered that the intermittent irrigation technique is also a promising approach to mitigate CH
                    <sub>4</sub> emissions by reductive soil conditions. Thus, integrating AWD and intermittent irrigation by incorporating biochar into the soil under the MD&#x2019;s edaphology, climate, and traditional practices could be feasible for further works.</p>
            </sec>
            <sec id="sec17">
                <title>Relationship between biochar amendment ratios and CH
                    <sub>4</sub> and N
                    <sub>2</sub>O fluxes</title>
                <p>There is a negative correlation between CH
                    <sub>4</sub> flux and RhB application rate (
                    <italic toggle="yes">P &lt;</italic> 0.001, 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.825) (
                    <xref ref-type="fig" rid="f4">Figure 4</xref>). It is indicated that CH
                    <sub>4</sub> flux decreased with the increase of rice-husk biochar application (
                    <xref ref-type="bibr" rid="ref66">Xiao 
                        <italic toggle="yes">et al.</italic> 2018</xref>). On the other hand, although increasing MB application rate could mitigate the CH
                    <sub>4</sub> emission, the relationship found a poor explanation (
                    <italic toggle="yes">P =</italic> 0.095, 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.254). This contrast could be partly attributed to biochar-carbonized properties. MB was low in the specific surface area and total pore volume compared to RhB (
                    <xref ref-type="table" rid="T1">Table 1</xref>). 
                    <xref ref-type="bibr" rid="ref21">Ji 
                        <italic toggle="yes">et al.</italic> (2020)</xref> revealed that biochar structure intimately related to anaerobic CH
                    <sub>4</sub> oxidation and created a suitable environment for CH
                    <sub>4</sub>-consuming bacteria.</p>
                <p>Similarly, we found a negative correlation between the N
                    <sub>2</sub>O flux reductions and the application rate of RhB (
                    <italic toggle="yes">P =</italic> 0.012, 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.619) and MB (
                    <italic toggle="yes">P =</italic> 0.002, 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.757). In agreement with our finding, a meta-analysis of 
                    <xref ref-type="bibr" rid="ref8">Cayuela 
                        <italic toggle="yes">et al.</italic> (2014)</xref> showed a negative relationship between biochar application rates and reduced N
                    <sub>2</sub>O flux, where sufficient N
                    <sub>2</sub>O reduction was 1-2% biochar amendments, whereas, incorporating more than 10% of biochar into the soil was found to reach up to 80%. In line with our study, 
                    <xref ref-type="bibr" rid="ref20">Huang 
                        <italic toggle="yes">et al.</italic> (2019)</xref> also showed a negative relationship between biochar application rates and N
                    <sub>2</sub>O flux. Overall, the increase of biochar application rates could potentially stimulate the CH
                    <sub>4</sub> and N
                    <sub>2</sub>O reduction. However, for CH
                    <sub>4</sub> and N
                    <sub>2</sub>O fluxes, the application of 5 and 10 t ha
                    <sup>&#x2212;1</sup> remains unclear.</p>
            </sec>
            <sec id="sec18">
                <title>Effect of biochar incorporation on Soil Eh and CH
                    <sub>4</sub> emission</title>
                <p>Our study found that the negative linear relationship between soil Eh and hourly CH
                    <sub>4</sub> flux with RhB (
                    <italic toggle="yes">P &lt;</italic> 0.001; 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.552) and MB (
                    <italic toggle="yes">P &lt;</italic> 0.001; 
                    <italic toggle="yes">r</italic>
                    <sup>2</sup> = 0.502) (
                    <xref ref-type="fig" rid="f5">Figure 5</xref>). Similar results were also observed by 
                    <xref ref-type="bibr" rid="ref58">Wang 
                        <italic toggle="yes">et al</italic>. (2018)</xref>. This indicates that an increase of soil redox potential decreased CH
                    <sub>4</sub> emission, which is in line with the report by 
                    <xref ref-type="bibr" rid="ref50">Towprayoon (2020)</xref>. Moreover, soil Eh remained below &#x2212;250 from 17 to 66 DAS in our study (
                    <xref ref-type="fig" rid="f4">Figure 4</xref>), implying a favorable condition for CH
                    <sub>4</sub> emission (
                    <xref ref-type="bibr" rid="ref60">Wang 
                        <italic toggle="yes">et al.</italic> 1993</xref>). Final drainage rapidly increased soil Eh and reduced CH
                    <sub>4</sub> flux (
                    <xref ref-type="fig" rid="f3">Figure 3</xref>), indicating the strong sensibility of soil Eh and CH
                    <sub>4</sub> flux under water management.</p>
                <p>Biochar application increased soil Eh compared to non-amended soils (
                    <xref ref-type="table" rid="T4">Table 4</xref>). This indicates that biochar was the critical factor contributing to the positive effects of anaerobic CH
                    <sub>4</sub> oxidation activities known as the electronic accepting capacities (EAC) of biochar (
                    <xref ref-type="bibr" rid="ref34">Nan 
                        <italic toggle="yes">et al.</italic>, 2021</xref>). The supplementation of biochar intensifies oxygen-containing functional groups (carboxyl, carbonyl, quinone phenolic hydroxyl group) and positively improves biochar redox potential (
                    <xref ref-type="bibr" rid="ref23">Kl&#x00fc;pfel 
                        <italic toggle="yes">et al.</italic> 2014</xref>; 
                    <xref ref-type="bibr" rid="ref62">Wu 
                        <italic toggle="yes">et al.</italic> 2016</xref>). The increase of Eh and the reduction of CH
                    <sub>4</sub> emissions could also be explained by the porosity and absorbability characteristics of biochar, which enable robust CH
                    <sub>4</sub>-utilizing bacteria activities and intensify the diffusion and metabolism process. In a similar way, biochar incorporation into soils improves soil aeration, creating a favorable environment for methanotrophic bacteria resulting in soil Eh amelioration and better reduction of CH
                    <sub>4</sub> oxidation (
                    <xref ref-type="bibr" rid="ref14">Feng 
                        <italic toggle="yes">et al.</italic>, 2012</xref>).</p>
            </sec>
            <sec id="sec19">
                <title>Effects of biochar incorporation on grain yield, GWP and Yield-scaled GWP</title>
                <p>Although biochar amendments could improve yield (2.41-4.21%) (
                    <xref ref-type="table" rid="T3">Table 3</xref>), multiple comparison analyses found no significant difference between amended and unamended soils. Several studies have found similar results (
                    <xref ref-type="bibr" rid="ref41">Qin 
                        <italic toggle="yes">et al.</italic> 2016</xref>; 
                    <xref ref-type="bibr" rid="ref35">Nguyen 
                        <italic toggle="yes">et al.</italic> 2016</xref>). The undistinctive grain yield could be partly attributed to spatial and temporal variations, i.e., climatic conditions, field practices, soil substrates (
                    <xref ref-type="bibr" rid="ref67">Xie 
                        <italic toggle="yes">et al.</italic> 2013</xref>).</p>
                <p>Biochar-amended soil significantly decreased GWP by 21.1-30.0% and yield-scaled GWP by 24.5% - 31.8% (
                    <xref ref-type="table" rid="T3">Table 3</xref>). It was indicated that RhB and MB application potentially mitigates total CH
                    <sub>4</sub> and N
                    <sub>2</sub>O emissions without scarifying grain yield. 
                    <xref ref-type="bibr" rid="ref68">Yang 
                        <italic toggle="yes">et al.</italic> (2019)</xref> performed a double-season field experiment on biochar applications ranging from 20 to 40 t ha
                    <sup>&#x2212;1</sup> and found that the average GWP and yield-scaled GWP reduced by 18.7% - 16.4%, and 80.3% - 41.6%, respectively. Similarly, Zhang 
                    <italic toggle="yes">et al.</italic> (2019) observed a six-year field experiment on biochar-applied soils at rates of between 20-40 t ha
                    <sup>&#x2212;1</sup> and showed a GWP and yield-scaled GWP reduction by 12.1-18.4% and 35.9-56.4%, respectively. Here we observed that CH
                    <sub>4</sub> flux was the key contributor in the GWP and yield-scaled GWP via the field experiment in the VMD&#x2019;s transition season, while N
                    <sub>2</sub>O flux was more neglectable. Thus, future works should emphasize on reducing the GWP, yield-scaled GWP, and concentrate on the CH
                    <sub>4</sub> mitigation technology solutions rather than N
                    <sub>2</sub>O emissions.</p>
            </sec>
            <sec id="sec20">
                <title>Effects of biochar incorporation on soil improvement</title>
                <p>Soil improvement under short-term and long-term biochar applications has been widely recognized. Our study showed that biochar amendment insignificantly increased soil pH (
                    <xref ref-type="table" rid="T4">Table 4</xref>), which indicated no effect of biochar addition on soil pH perfection as suggested by previous studies (
                    <xref ref-type="bibr" rid="ref68">Yang 
                        <italic toggle="yes">et al.</italic>, 2019</xref>). However, biochar amendment significantly decreased the soil bulk density and improved soil porosity in comparison to non-amended soils. Furthermore, higher applied biochar rates showed lesser soil bulk density and higher porosity indicating that biochar directly upgraded soil physiology. Amelioration of soil surface area and porosity by biochar amendment intensifies soil aeration and functions of aeration, such as CH
                    <sub>4</sub> oxidation, and provides habitat for methanotrophs (
                    <xref ref-type="bibr" rid="ref34">Nan 
                        <italic toggle="yes">et al</italic>., 2021</xref>). Moreover, it stimulates NH
                    <sub>4</sub>
                    <sup>+</sup> absorbance ability resulting in suppressing nitrification processes and N
                    <sub>2</sub>O flux reduction in the field (
                    <xref ref-type="bibr" rid="ref59">Wang 
                        <italic toggle="yes">et al.</italic>, 2020</xref>).</p>
                <p>It is evident that increasing biochar application boosted SOM and AN, with a slightly increased available P through the season (
                    <xref ref-type="table" rid="T4">Table 4</xref>). The increasing of SOM and AN showed a high nutrient availability in the soil. Notably, the soil improvement did not increase soil CH
                    <sub>4</sub> and N
                    <sub>2</sub>O emissions as above-mentioned and discussed. AN could be used as a soil health indicator (
                    <xref ref-type="bibr" rid="ref16">Garc&#x00ed;a 
                        <italic toggle="yes">et al.</italic>, 2020</xref>). The interdependence among AN, SOC, and particulate OC was demonstrated by a positive correlation (
                    <xref ref-type="bibr" rid="ref13">Dom&#x00ed;nguez 
                        <italic toggle="yes">et al.</italic>, 2016</xref>). In connection with our study, 
                    <xref ref-type="bibr" rid="ref68">Yang 
                        <italic toggle="yes">et al</italic>. (2019)</xref> observed that biochar amendment slightly increased SOC, significantly increased NH
                    <sub>4</sub>
                    <sup>+</sup> by 47.7%, and significantly decreased NO
                    <sub>3</sub>
                    <sup>&#x2212;</sup> by 30.4%. Incorporating biochar into soils could inhibit nitrification and produce more NH
                    <sub>4</sub>
                    <sup>+</sup> than NO
                    <sub>3</sub>
                    <sup>&#x2212;</sup> consisting of an anoxic environment (water level and redox potential; 
                    <xref ref-type="fig" rid="f2">Figure 2</xref>). Increasing NH
                    <sub>4</sub>
                    <sup>+</sup> concentrations and declining NO
                    <sub>3</sub>
                    <sup>&#x2212;</sup> concentrations would partly explain the enhanced CH
                    <sub>4</sub>-consuming figure and N
                    <sub>2</sub>O oxidation (
                    <xref ref-type="bibr" rid="ref66">Xiao 
                        <italic toggle="yes">et al.</italic>, 2018</xref>). Overall, biochar application offers benefits not only for nutrients availability, but also for GHGs mitigation.</p>
            </sec>
        </sec>
        <sec id="sec21" sec-type="conclusions">
            <title>Conclusions</title>
            <p>This study assessed the effects of rice husk biochar or melaleuca biochar amendment at 5 or 10 t ha
                <sup>&#x2212;1</sup> on CH
                <sub>4</sub> and N
                <sub>2</sub>O emissions and the physiochemical soil properties after rice cultivation under typical water management and conventional practice regime in the VMD. Incorporating biochar into soils significantly mitigated CH
                <sub>4</sub> and N
                <sub>2</sub>O emissions without reducing grain yield. Consequently, a lower GWP and yield-scaled GWP from biochar-amended soils were achieved. Although higher biochar applications decreased CH
                <sub>4</sub> and N
                <sub>2</sub>O emissions, there was no significant difference between biochar-amended rates. Biochar significantly increased soil Eh conditions. There was a negative linear relationship between soil Eh and CH
                <sub>4</sub> emission rate for biochar-applied fields. N
                <sub>2</sub>O emissions from biochar fields were relatively low and mostly concentrated during the fertilization period. Biochar amendments improved soil fertility via physical properties of soils by decreasing bulk density and intensifying porosity and the chemical characteristics of the soils by ameliorating SOM, AN and AP, but did not affect soil pH. Similar to GHG emissions, biochar application rates of between 5 and 10 t ha
                <sup>&#x2212;1</sup> could not obtain significant soil improvement. This study will help lower-GHG emissions from rice farming practices in the VMD. Further works should study the combination of biochar-application rates and effective water irrigation techniques on different soils in the VMD.</p>
        </sec>
        <sec id="sec22">
            <title>Data availability</title>
            <sec id="sec23">
                <title>Underlying data</title>
                <p>Figshare: Biochar reduces GHGs from paddy fields. 
                    <ext-link ext-link-type="uri" xlink:href="https://doi.org/10.6084/m9.figshare.16625137.v1">https://doi.org/10.6084/m9.figshare.16625137.v1</ext-link> (
                    <xref ref-type="bibr" rid="ref51">Tran Sy 
                        <italic toggle="yes">et al</italic>., 2021</xref>).</p>
                <p>This project contains the following underlying data:
                    <list list-type="bullet">
                        <list-item>
                            <label>-</label>
                            <p>Nam et al_Raw data biochar_F1000research.xlsx</p>
                        </list-item>
                    </list>
                </p>
                <p>Data are available under the terms of the 
                    <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/publicdomain/zero/1.0/">Creative Commons Zero &#x201c;No rights reserved&#x201d; data waiver</ext-link> (CC0 1.0 Public domain dedication).</p>
            </sec>
        </sec>
    </body>
    <back>
        <ack>
            <title>Acknowledgments</title>
            <p>This study is funded in part by the Can Tho University Improvement Project VN14-P6, supported by a Japanese ODA loan. We would like to thank Cuu Long Delta Rice Research Institute (CLRRI), Vietnam, and Mr. Ho Minh Nhut (master student K25 in Can Tho University, Vietnam) for their support of this study. We also thank Dr. Nigel Downes for proofreading the manuscript</p>
        </ack>
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    <sub-article article-type="reviewer-report" id="report120157">
        <front-stub>
            <article-id pub-id-type="doi">10.5256/f1000research.77750.r120157</article-id>
            <title-group>
                <article-title>Reviewer response for version 1</article-title>
            </title-group>
            <contrib-group>
                <contrib contrib-type="author">
                    <name>
                        <surname>Tho</surname>
                        <given-names>Bui Truong</given-names>
                    </name>
                    <xref ref-type="aff" rid="r120157a1">1</xref>
                    <role>Referee</role>
                </contrib>
                <aff id="r120157a1">
                    <label>1</label>The Centre of Hi-tech Application in Agriculture, Chau Thanh district, Vietnam</aff>
            </contrib-group>
            <author-notes>
                <fn fn-type="conflict">
                    <p>
                        <bold>Competing interests: </bold>No competing interests were disclosed.</p>
                </fn>
            </author-notes>
            <pub-date pub-type="epub">
                <day>8</day>
                <month>2</month>
                <year>2022</year>
            </pub-date>
            <permissions>
                <copyright-statement>Copyright: &#x00a9; 2022 Tho BT</copyright-statement>
                <copyright-year>2022</copyright-year>
                <license xlink:href="https://creativecommons.org/licenses/by/4.0/">
                    <license-p>This is an open access peer review report distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
                </license>
            </permissions>
            <related-article ext-link-type="doi" id="relatedArticleReport120157" related-article-type="peer-reviewed-article" xlink:href="10.12688/f1000research.74041.1"/>
            <custom-meta-group>
                <custom-meta>
                    <meta-name>recommendation</meta-name>
                    <meta-value>approve</meta-value>
                </custom-meta>
            </custom-meta-group>
        </front-stub>
        <body>
            <p>This article, "Rice husk and Melaleuca biochar additions reduce soil CH
                <sub>4</sub>&#x00a0;and N
                <sub>2</sub>O emissions and increase soil organic matter and nutrient availability" aims to evaluate the impacts of two biochar amendments on soil CH
                <sub>4</sub> and N
                <sub>2</sub>O emissions and its effect on soil physiochemical properties in Mekong Delta based on 80 days of field study.</p>
            <p> </p>
            <p> The topic is interesting and poorly treated in literature. The data are correctly processed and the results are well discussed. However, there are some minor issues and my reviews are focused on improving a revision of the current article, and I hope the authors can accommodate my suggestions. 
                <list list-type="order">
                    <list-item>
                        <p>Site description: &#x201c;air temperature (27.5 &#x2013; 27.5 &#x00b0;C)&#x201d; I think there is a mistake, authors should double-check the information.</p>
                    </list-item>
                    <list-item>
                        <p>Statistical analysis: Authors should confirm that all data were test homogeneity of variance for using One-Way analysis of variance (ANOVA).</p>
                    </list-item>
                    <list-item>
                        <p>Authors used &#x201c;Vietnamese Mekong Delta (VMD)&#x201d;, some places used &#x201c;Mekong Delta (MD)&#x201d;, it should unified in all the manuscript.</p>
                    </list-item>
                </list>
            </p>
            <p>Is the work clearly and accurately presented and does it cite the current literature?</p>
            <p>Yes</p>
            <p>If applicable, is the statistical analysis and its interpretation appropriate?</p>
            <p>Partly</p>
            <p>Are all the source data underlying the results available to ensure full reproducibility?</p>
            <p>Partly</p>
            <p>Is the study design appropriate and is the work technically sound?</p>
            <p>Yes</p>
            <p>Are the conclusions drawn adequately supported by the results?</p>
            <p>Yes</p>
            <p>Are sufficient details of methods and analysis provided to allow replication by others?</p>
            <p>Yes</p>
            <p>Reviewer Expertise:</p>
            <p>Hi-tech Application in Agriculture, Climate change mitigation, GHG emissions, Sustainable crop rotations, Plant ecophysiology.</p>
            <p>I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.</p>
        </body>
        <sub-article article-type="response" id="comment7792-120157">
            <front-stub>
                <contrib-group>
                    <contrib contrib-type="author">
                        <name>
                            <surname>Huynh Van</surname>
                            <given-names>Thao</given-names>
                        </name>
                        <aff>College of Environment and Natural Resources, Cantho Univeristy, Vietnam</aff>
                    </contrib>
                </contrib-group>
                <author-notes>
                    <fn fn-type="conflict">
                        <p>
                            <bold>Competing interests: </bold>No competing interests were disclosed.</p>
                    </fn>
                </author-notes>
                <pub-date pub-type="epub">
                    <day>9</day>
                    <month>2</month>
                    <year>2022</year>
                </pub-date>
            </front-stub>
            <body>
                <p>Thank you very much for your constructive comments. All your comments have been carefully read and revised by the authors. Our responses are enclosed as follows:</p>
                <p> </p>
                <p> 
                    <bold>Comment 1#.</bold> Site description: &#x201c;air temperature (27.5 &#x2013; 27.5 &#x00b0;C)&#x201d; I think there is a mistake, authors should double-check the information.</p>
                <p> 
                    <bold>
                        <italic>Reply 1:</italic>
                    </bold>&#x00a0;Thank you for finding this mistake. We re-referred to the original document and corrected the information in the manuscript. The air temperature should be 27.5
                    <sup>o</sup>C instead.</p>
                <p> </p>
                <p> 
                    <bold>Comment 2#:</bold> Statistical analysis: Authors should confirm that all data were test homogeneity of variance for using One-Way analysis of variance (ANOVA).</p>
                <p> 
                    <bold>
                        <italic>Reply 2:</italic>
                    </bold> We amended additional information related to the homogeneity of variance in the statistical analysis. The revised is as follows &#x201c;Significant different comparison among treatments was considered at Duncan&#x2019;s multiple range test (***P &lt; 0.001, **P &lt; 0.01, *P &lt; 0.05 and &#x2020;P &gt; 0.05) after passing homogeneity of variance&#x201d;.</p>
                <p> </p>
                <p> 
                    <bold>Comment 3:</bold> Authors used &#x201c;Vietnamese Mekong Delta (VMD)&#x201d;, some places used &#x201c;Mekong Delta (MD)&#x201d;, it should be unified in all the manuscript.</p>
                <p> 
                    <bold>
                        <italic>Reply 3:</italic>
                    </bold> We have unified the abbreviation in all parts of the manuscript which is &#x201c;VMD&#x201d;.</p>
                <p> </p>
                <p> Best regards,</p>
                <p> Authors.</p>
            </body>
        </sub-article>
    </sub-article>
    <sub-article article-type="reviewer-report" id="report99426">
        <front-stub>
            <article-id pub-id-type="doi">10.5256/f1000research.77750.r99426</article-id>
            <title-group>
                <article-title>Reviewer response for version 1</article-title>
            </title-group>
            <contrib-group>
                <contrib contrib-type="author">
                    <name>
                        <surname>Tariq</surname>
                        <given-names>Azeem</given-names>
                    </name>
                    <xref ref-type="aff" rid="r99426a1">1</xref>
                    <role>Referee</role>
                    <uri content-type="orcid">https://orcid.org/0000-0002-6132-2391</uri>
                </contrib>
                <aff id="r99426a1">
                    <label>1</label>Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark</aff>
            </contrib-group>
            <author-notes>
                <fn fn-type="conflict">
                    <p>
                        <bold>Competing interests: </bold>No competing interests were disclosed.</p>
                </fn>
            </author-notes>
            <pub-date pub-type="epub">
                <day>4</day>
                <month>1</month>
                <year>2022</year>
            </pub-date>
            <permissions>
                <copyright-statement>Copyright: &#x00a9; 2022 Tariq A</copyright-statement>
                <copyright-year>2022</copyright-year>
                <license xlink:href="https://creativecommons.org/licenses/by/4.0/">
                    <license-p>This is an open access peer review report distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
                </license>
            </permissions>
            <related-article ext-link-type="doi" id="relatedArticleReport99426" related-article-type="peer-reviewed-article" xlink:href="10.12688/f1000research.74041.1"/>
            <custom-meta-group>
                <custom-meta>
                    <meta-name>recommendation</meta-name>
                    <meta-value>approve-with-reservations</meta-value>
                </custom-meta>
            </custom-meta-group>
        </front-stub>
        <body>
            <p>The authors of "Rice husk and melaleuca biochar additions reduce soil CH
                <sub>4</sub> and N
                <sub>2</sub>O emissions and increase soil organic matter and nutrient availability" present an effort in assessing the effects of two biochar amendments on soil CH
                <sub>4</sub> and N
                <sub>2</sub>O emissions and effects on soil physiochemical properties based on 80 days of field study. The overall paper has been written in a good English and in a logical manner. However, there are some minor issues that authors need to deal before considering this paper for indexing.</p>
            <p> </p>
            <p> Authors used the term &#x201c;soil organic matter and nutrient availability&#x201d; in title, but they have focused on different soil properties e.g. soil porosity, soil bulk density, soil redox potential, soil organic matter and nitrogen. Authors should use the term soil physiochemical properties instead.</p>
            <p> </p>
            <p> Authors did not explain the results related to soil properties in the abstract section (e.g. there is no explanation about soil organic matter and nitrogen contents in abstract).</p>
            <p> </p>
            <p> Authors stated that they used 95 % confidence level, but the result section predicted that authors have used different levels of probability for difference analysis. Author should explain the analysis carefully in the statistical analysis section in Material and Methods.</p>
            <p>Is the work clearly and accurately presented and does it cite the current literature?</p>
            <p>Yes</p>
            <p>If applicable, is the statistical analysis and its interpretation appropriate?</p>
            <p>Partly</p>
            <p>Are all the source data underlying the results available to ensure full reproducibility?</p>
            <p>Partly</p>
            <p>Is the study design appropriate and is the work technically sound?</p>
            <p>Yes</p>
            <p>Are the conclusions drawn adequately supported by the results?</p>
            <p>Yes</p>
            <p>Are sufficient details of methods and analysis provided to allow replication by others?</p>
            <p>Yes</p>
            <p>Reviewer Expertise:</p>
            <p>Climate change mitigation, GHG emissions, sustainable crop rotations, ecosystem modeling</p>
            <p>I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.</p>
        </body>
        <sub-article article-type="response" id="comment7791-99426">
            <front-stub>
                <contrib-group>
                    <contrib contrib-type="author">
                        <name>
                            <surname>Huynh Van</surname>
                            <given-names>Thao</given-names>
                        </name>
                        <aff>College of Environment and Natural Resources, Cantho Univeristy, Vietnam</aff>
                    </contrib>
                </contrib-group>
                <author-notes>
                    <fn fn-type="conflict">
                        <p>
                            <bold>Competing interests: </bold>No competing interests were disclosed.</p>
                    </fn>
                </author-notes>
                <pub-date pub-type="epub">
                    <day>9</day>
                    <month>2</month>
                    <year>2022</year>
                </pub-date>
            </front-stub>
            <body>
                <p>Thank you very much for giving us constructive comments that helped us improve this paper immensely. All your comments have been carefully read and revised to elucidate underlying aspects. Our responses to your comments are as follows:</p>
                <p> </p>
                <p> 
                    <bold>Comment 1:</bold> Authors used the term &#x201c;soil organic matter and nutrient availability&#x201d; in title, but they have focused on different soil properties e.g. soil porosity, soil bulk density, soil redox potential, soil organic matter and nitrogen. Authors should use the term soil physiochemical properties instead.</p>
                <p> 
                    <bold>
                        <italic>Reply 1#:</italic>
                    </bold> Thank you very much for the recommendation for the title term of &#x201c;soil physiochemical properties&#x201d;. The recommendation has been adopted. The revision is as follows &#x201c;Rice husk and melaleuca biochar additions reduce soil CH
                    <sub>4</sub> and N
                    <sub>2</sub>O emissions and increase soil physicochemical properties.&#x201d;</p>
                <p> </p>
                <p> 
                    <bold>Comment 2:</bold> Authors did not explain the results related to soil properties in the abstract section (e.g. there is no explanation about soil organic matter and nitrogen contents in abstract).</p>
                <p> 
                    <bold>
                        <italic>Reply 2#:</italic>
                    </bold> We revised the abstract with additional information related to the explanation and implication for improving soil physiochemical characteristics. The revision is as follows &#x201c;The results showed&#x2026;Ameliorating soil aeration and functions by adding RhB and MB resulted in improving soil physicochemical characteristics, especially significant SOM and AN boosting, which indicate better soil health, structure, and fertility&#x201d;.</p>
                <p> </p>
                <p> 
                    <bold>Comment 3:</bold> Authors stated that they used 95% confidence level, but the result section predicted that authors have used different levels of probability for difference analysis. Author should explain the analysis carefully in the statistical analysis section in Material and Methods.</p>
                <p> 
                    <bold>
                        <italic>Reply 3#:</italic>
                    </bold> We added more information to the statistical level to make it clearer to readers. The supplementations are as follows &#x201c;The results are&#x2026;Significant different comparison among treatments was considered at Duncan&#x2019;s multiple range test (***P &lt; 0.001, **P &lt; 0.01, *P &lt; 0.05 and &#x2020;P &gt; 0.05)&#x201d;</p>
                <p> </p>
                <p> We look forward to hearing from you.</p>
                <p> Best regards,</p>
                <p> Authors.</p>
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