The complete genome sequences of Penicillium concavorugulosum

Introduction

Penicillium species are a widespread group of facultative fungi which are found in various habitats such as soil, air and decaying plant matter (Pitt and Hocking, 2009; Visagie et al., 2005). Various studies have reported the genus to contain several species which play different roles in agriculture and human health. For example, species such as Penicillium cyclopium and Penicillium citrinum have been reported to be contaminants in corn, soybean, and dried beans (Munkvold et al., 2019). Whereas Penicillium expansum and many other Penicillium species have been reported to cause post-harvest losses in pome fruits and apples (Pitt and Hocking, 2009; Wu et al., 2019).

Many Penicillium species produce extracellular enzymes such as cellulolytic enzymes, polysaccharases and pectic enzymes that aid in the breakdown of organic materials and use in antibiotic production, cell degeneration, as well as food production (Yoon et al., 2007; Rabha and Jha, 2018). Production of these enzymes and phytohormones by Penicillium fungal species that influence plant physiological processes was also reported by (Chanclud and Morel, 2016). Tiwari et al. (2011) also highlighted Penicillium species as some of many fungal microorganisms that possess multifunctional properties that enable them to be used in various agro-industries, biological, medicinal, and commercial purposes.

This potential widespread use of different fungal species, including Penicillium species, require accurate identification. The need for accurate identification of various important micro-organisms has led to the development and utilization of molecular identification techniques. This technology uses polymerase chain reaction (PCR). Fungal microorganisms are mostly analyzed using the PCR method to determine the ITS region for accurate differentiation of species amongst the genus (Johnston, 2011).

One of the three Penicillium fungal species was established to possess multifunctional properties such as antibiosis, competition and myco-parasitism, and therefore was considered valuable for use as a bio-control agent against soil fungal pathogens (Segone, 2021). Furthermore, complete genome sequences for this Penicillium fungal species would also contribute to taxonomic studies, and evolution processes as well as understanding its various inherent antagonistic properties.

Methods

Four Penicillium species were isolated by use of soil serial dilution procedure. Soil was collected from different sites at the North-West University Mafikeng agricultural farm (25.8278° S, 25.6079° E). These sites had previously been subjected to different agronomic practices such as minimum soil tillage, weed control, fertilizer management and controlled irrigation.

DNA was extracted from the isolate mycelium by use of the Quick-DNA fungal/bacterial Miniprep Kit (Zymo Research, Catalogue No. D6005) (White et al., 1990).

Amplification of the target genes was carried out by use of OneTaq Quick load 2×Master Mix (NEB, Catalogue No. M0486) with primers presented in Table 1. Each Eppendorf tube comprised 10μL of NEB OneTaq 2X MasterMix with Standard Buffer (Catalogue No. M0482S), 1μL of genomic DNA (10-30ng/μL), 1μL forward primer (10μM), 1μL reverse primer (10μM), and 7μL nuclease-free water (Catalogue No. E476). Amplification cycles had an initial denaturation at 94°C for 10 minutes, 35 cycles of denaturation at 94°C for 30 seconds, annealing at 50°C, elongation at 68°C for 1 minute and final elongation at 72°C for 10 minutes. The PCR amplicons were stored at 4°C until electrophoresis.

Results

The genome assembly for the test Penicillium fungal species yielded a total sequence length of 570 with an N50 value of 796.6 bits (882), Expect = 0E00, Identities = 441/441 (100%), Gaps = 0/441 (0%).

This Penicillium fungal isolate was identified as Penicillium concavorugulosum (Accession: MK 841454.1).