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Article

Genomic Sequence Resource of Talaromyces albobiverticillius, the Causative Pathogen of Pomegranate Pulp Rot Disease

by
Tan Wang
1,*,
Shuchang Chen
1,
Qiuhong Niu
1,
Guangling Xu
1,
Chenxu Lu
1 and
Jin Zhang
2
1
School of Life Science and Agricultural Engineering, Nanyang Normal University, Nanyang 473061, China
2
School of Forestry and Biotechnology, Zhejiang A&F University, Hangzhou 311300, China
*
Author to whom correspondence should be addressed.
J. Fungi 2023, 9(9), 909; https://doi.org/10.3390/jof9090909
Submission received: 27 July 2023 / Revised: 2 September 2023 / Accepted: 6 September 2023 / Published: 7 September 2023
(This article belongs to the Special Issue Bioinformatics in Pathogenic Fungi)
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Abstract

:
Talaromyces albobiverticillius, a prominent pathogen responsible for pomegranate pulp rot disease, inflicts significant damage on Punica granatum L. Besides its pathogenicity, this fungus possesses the potential to produce substantial amounts of red pigments, making it promising for industrial applications. This study presents the genome annotation of T. albobiverticillius field strain Tp-2, isolated from pomegranates. The genome assembly, generated through a combination of Oxford Nanopore and Illumina sequencing reads, yielded a high-quality assembly with 14 contigs, featuring an N50 length of 4,594,200 bp. The complete genome of strain Tp-2 spans 38,354,882 bp, with a GC content of 45.78%. Importantly, the assembly exhibits remarkable integrity, with 98.3% of complete Benchmarking Universal Single-Copy Orthologs validating genome completeness. Genome prediction analysis reveals the presence of 10,380 protein-coding genes. To our knowledge, this study is the first report on the genome sequence of T. albobiverticillius, offering valuable insights into its genetic variation and molecular mechanisms of pigment production.

1. Introduction

The genus Talaromyces was first described by Benjamin in 1955 as a sexual state of Penicillium [1]. In 2011, Professors Houbraken and Samson redefined the genus Talaromyces based on genetic identification and the priority principle of international nomenclature regulations [2]. The number of Talaromyces species is rapidly increasing, owing to the availability of reliable and complete sequence data contributed from around the world [3]. Talaromyces includes medically important species, such as Talaromyces marneffei, which are associated with acquired immunodeficiency syndrome caused by human immunodeficiency virus infections [4]. Moreover, Talaromyces comprises species known for their production of various bioactive compounds, including anticancer agents [5], antibacterial and antifungal compounds [6], antioxidative compounds [7], enzymes [8,9], and natural colorants [10].
Among these pigment-producing species, Talaromyces albobiverticillius generates a substantial number of nontoxic Monascus-like azaphilone red pigments [11,12,13] which can be used as natural colorants in industrial applications. Meanwhile, pomegranate fruit pulp rot disease caused by T. albobiverticillius has been found in Italy [14] and China [15], resulting in significant production losses. The cultivation of pomegranate (Punica granatum L.) has recently sparked growing interest due to its potential impact on human health. Losses experienced both during cultivation and after harvesting are the main concerns in this valuable production chain. The main postharvest diseases of pomegranates were caused by latent pathogens that infect the fruits during the blooming phase (Coniella granati, Alternaria spp., Botrytis spp.), and secondarily by wound pathogens (Penicillium spp., Talaromyces spp., and Aspergillus spp.) that affect the fruits during the processing from harvest until the conclusion of storage [16,17]. In 2020, instances of postharvest disease in pomegranates attributed to Talaromyces spp. were documented in Serbia [18]. Hence, within the pomegranate sector, investigating the genomic genetic data and the pathogenic mechanisms of Talaromyces spp. has become highly imperative.
However, no information on the T. albobiverticillius genome is currently available. Herein, we present the first genome assembly for T. albobiverticillius and analyze its genomic features using Oxford Nanopore and Illumina sequencing technologies. The availability of this genomic data is likely to aid research on evolution, pigment manufacturing systems, infection mechanisms, and disease management.

2. Materials and Methods

2.1. Fungal Material and Culture Conditions

The strain Tp-2 used in this study was originally isolated from contaminated pomegranate fruits gathered from Xichuan, a city in China’s Henan province. The surface of the colony was grayish-green with a claret-red exudate. Fuchsia pigments are normally secreted into potato dextrose agar (PDA) medium. T. albobiverticillius was identified as the strain Tp-2 based on morphological traits and a multigene phylogenetic analysis [15]. Before DNA extraction, the strain was grown on PDA medium for 5 days at 25 °C. Mycelium strain Tp-2 was harvested and immediately frozen in liquid nitrogen for genomic DNA extraction.

2.2. DNA Extraction and Genome Sequencing

The genomic DNA of the Tp-2 strain was extracted from 5-day-old fresh mycelia using the sodium dodecyl sulfate method [19]. Briefly, 1 g of fresh mycelia was collected and combined with 900 μL of 10% SDS lysis buffer. To this mixture, 100 μg proteinase K per ml of emulsion solution was added, and the solution was incubated for 1 h at 50 °C. Subsequently, 350 μL of neutral saturated salt solution (NaCl) per ml was introduced into the tube. The tube was then centrifuged at 590× g for 15 min, resulting in DNA remaining exclusively in the aqueous phase. The resultant aqueous phase was mixed with two volumes of absolute ethyl alcohol. After removing the supernatant, the DNA pellet was washed once with 75% ethanol and the DNA was precipitated out of the solution by centrifugation at 9500× g for 5 min. The isolated DNA was detected using 1% agarose gel electrophoresis and quantified using the Qubit 2.0 Fluorometer (Thermo Scientific, Waltham, MA, USA). Libraries for nanopore sequencing were constructed with a 10-kb insert size of genomic DNA. First, large DNA fragments were recovered using the Blue Pippin automatic nucleic acid fragment recovery system and then repaired. Barcodes were added using the PCR-free method of the EXP-NBD104 kit (Oxford Nanopore Technologies, Oxford, UK). The size of the fragments was subsequently determined using the AATI automatic mixer (Agilent Technologies, Santa Clara, CA, USA). Finally, the SQK-LSK109 connection kit (Oxford Nanopore Technologies, Oxford, UK) was used to link the adapter, completing the construction of the 10-K library. Libraries for Illumina sequencing were generated using the NEBNext® UltraTM DNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA, USA) following the manufacturer’s recommendations, with index codes added to attribute sequences to each sample. Briefly, DNA samples were sonicated to a size of 350 bp, and then DNA fragments were end-polished, A-tailed, and ligated with full-length adapters for Illumina sequencing, followed by further PCR amplification. The PCR products were purified using AMPure XP Bead-based reagent (Beckman, Indianapolis, IN, USA), and the libraries were analyzed for size distribution using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) and quantified using real-time PCR. The whole genome of the Tp-2 strain was sequenced using the Nanopore PromethION 48 platform and Illumina NovaSeq PE150 (Novogene Bioinformatics Technology, Beijing, China).

2.3. Genome Assembly and Prediction

The nanopore data was assembled using the Flye software (version 2.9-b1768), which produced preliminary assembly results. The splicing results were corrected using Racon (version 1.4.13) and Pilon (version 1.22) software, which produced final assembly results. The Benchmarking Universal Single-Copy Orthologs (BUSCO version 3.0.2) assessment used a single-copy orthologous gene library, and software such as Metaeuk and Hmmer were used to evaluate the assembled genome to assess the assembled genome’s integrity. The Augustus (version 2.7) program was used to retrieve predicted coding genes.

2.4. Phylogenetic Analysis

The NCBI Genome Database was used to retrieve the annotated gene sequences from additional species of phylogenetically related Talaromyces taxa. Genome alignments were performed in MUMmer 3 (version 3.2.2) using an all-against-all comparison with the default parameters. Then, the tool OrthoFinder (version 2.4.0) was used to identify key orthologs [20]. With the help of IQ-Tree (version 2.2.0), a phylogenetic tree was constructed [21].

2.5. Dispersed Repeat Sequences and Noncoding tRNA Annotation

Dispersed repeat sequences were masked throughout the genome using the Repeat Masking option (RepeatMasker v.4.0.5) [22], which was implemented in OmicsBox software v.1.4.12. Tandem repeats were analyzed using TRF (version 4.07b) [23]. Transfer RNA (tRNA) genes were predicted using tRNAscan-SE (version 1.3.1) [24], while ribosomal RNA (rRNA) genes were analyzed using rRNAmmer (version 1.2) [25]. For small RNA (sRNA) analysis, we initially aligned and annotated the data with the Rfam database [26]. Subsequently, the CMsearch program (Version 1.1rc4) was employed to determine the final sRNA annotations.

2.6. Gene Annotation and Functional Analyses

Basic annotations of predicted genes were performed using seven major databases, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Clusters of Orthologous Groups (KOG), Non-Redundant Protein Database (NR), Transporter Classification Database (TCDB), Pfam, and Swiss-Prot [27,28,29,30,31]. We comprehensively predicted whether the protein sequence was a secreted protein using SignalP (Version 4.1) [32] and TMHMM (Version 2.0c) algorithms to determine whether the genome sequences included signal peptides and transmembrane structures. Meanwhile, we analyzed the secondary metabolism gene clusters using antiSMASH (version 5.1) [33] and causative genes using Pathogen–Host Interactions (PHI, version 4.12) and Database of Fungal Virulence Factors (DFVF) [34]. The Carbohydrate-Active Enzymes (CAZymes) Database [35] predicted CAZymes.

3. Results

3.1. Genome Assembly and Genomic Characteristics

We present the first de novo genome sequencing of the Tp-2 strain of T. albobiverticillius using short- and long-read technologies. In this study, nanopore sequencing produced 6,588,890,037 bp of raw read data and Illumina paired-end sequencing produced 4844 Mb of raw read data (described in Section 2.2). The assembled Tp-2 genome had a length of 38,354,882 bp and a GC content of 45.78%. The BUSCO analysis (described in Section 2.3) showed that 98.3% of the BUSCOs were complete and that 98.3%, 0%, and 0.3% of the BUSCOs were single-copy, duplicated, and fragmented, respectively (Figure S1). More characteristics of the assembled genome are presented in Table 1 and Figure 1.
The assembly statistics of T. albobiverticillius and its phylogenetically related species are presented in Table 2. To establish the evolutionary relationship among the Talaromyces species, pairwise comparisons were conducted using the genomes of T. albobiverticillius, Tp-2, T. atroroseus IBT 11181, T. islandicus WF-38-12, T. marneffei ATCC 18224, and T. verruculosus TS63-9 (described in Section 2.4). These assessments revealed 3777 single-copy orthologous genes shared among these five species. Additionally, the genome data of T. atroroseus, T. islandicus, T. marneffei, and T. verruculosus contained 6396, 5809, 5871, and 6493 one-to-one orthologous genes, respectively. The phylogenetic analysis illustrated that T. atroroseus exhibited a closer evolutionary relationship to T. albobiverticillius than to other species (Figure 2).

3.2. Gene Prediction and Functional Annotation

We employed the Augustus 2.7 program to predict fungal genes and obtained 10,380 genes (Table 3). Subsequently, through rRNAmmer and tRNAscan-SE analyses, we predicted 204 noncoding RNAs, which included 47 ribosomal RNAs (rRNA) and 157 transfer RNAs (tRNA) (described in Section 2.5). Our investigation led to the identification of 7412 proteins harboring Pfam domains, 8844 genes participating in various KEGG pathways, and 2160 KOG genes (described in Section 2.6). Additionally, antiSMASH analysis revealed 214 Type I polyketide synthases (T1PKSs) genes closely associated with red pigment synthesis in T. albobiverticillius. Furthermore, we mapped 750 genes to the CAZyme database and encoded CAZymes. A total of 227 genes were assigned to the cytochrome P450 family. Notably, 400 virulence genes associated with pathogenesis were predicted via DFVF. Additionally, our search of the PHI database revealed 1332 associated genes. Moreover, we identified 2057 transmembrane proteins, 638 transport proteins, and 671 secreted proteins using the TMHMM, TCDB, and SignalP databases, respectively. These genes are involved in the pathogenesis of fungi and may play an important role in the pathogenesis of T. albobiverticillius.

3.2.1. GO Annotation

The database categorizes genes into three groups based on their roles, including cellular component, molecular function, and biological process (described in Section 2.6). Gene function classification analysis of T. albobiverticillius revealed that most genes participating in biological processes were involved in metabolic (3988), cellular (2971), and single-organism (2657) activities, as well as localization (1491) processes (Table S1). Cell (2806), membrane (2589), organelle (1444) portions, macromolecular complex (646), and membrane-enclosed lumen (139) were the most common gene categories listed in the cellular component category. Genes were mostly involved in catalytic activity (4117), binding (3660), transporter activity (785), structural molecule activity (128), and signal transducer activity (54).

3.2.2. Annotation of CAZymes

The CAZymes database is a professional database related to carbohydrate enzymes, including those capable of catalyzing carbohydrate degradation and modification as well as related enzyme families for biosynthesis (described in Section 2.6). A total of 750 genes encoding putative CAZymes were identified in T. albobiverticillius (Table S2). Under the five categories and one structural domain, multiple families have been established. Based on the results, GHs were the most prevalent family, with 105 enzymes (427 genes). The next most abundant category was GTs, with 37 enzymes (121 genes), followed by the AA family with 14 enzymes (60 genes). The other three categories, namely CBMs, CEs, and PLs, were relatively less numerous, containing 12 (91 genes), 10 (40 genes), and 8 (11 genes) enzymes, respectively (Table 4). The main GH subfamilies detected were chitinases (GH18), β-glucosidases (GH3), polygalacturonases (GH28), invertase (GH32), α-L-rhamnosidase (GH78), α-1,6-mannanase (GH76), α-glucosidase (GH31), α-galactosidase (GH27), and α-1,2-mannosyltransferase (GH22), which are listed in descending order of the quantity of genes. Within the GT families, the most abundant were those involved in N-acetyllactosaminide (GT31), UDP-glucuronosyltransferases (GT1), α-1,2-mannosyltransferase (GT22), and α-trehalose-phosphate synthase (GT20). Among the AA family, the most predominant subfamilies were oxidoreductases (AA1) and glucooligosaccharide oxidases (AA7). Regarding the CE subfamilies, acetylesterase (CE16) and acetyl xylan esterase (CE5) were the most abundant, while the chitopentaose-binding (CBM50) family was the most prominent in the CBM category.

3.2.3. Annotation of Transport Proteins

The Transporter Classification Database (TCDB) is a classification system for predicting membrane transport proteins (described in Section 2.6). In T. albobiverticillius, 638 genes encoding transport proteins were discovered, with 18 genes (2.8%) sharing more than 80% similarity (Table S3). The electrochemical potential-driven transporters (263 genes) were the most prominent group, followed by the main active transporters (163 genes) and channels and pores (73 genes).

3.2.4. Annotation of Secreted Proteins

Secreted proteins are those that are produced within a cell and are subsequently released to exert their effects outside of it. The N-terminus of secreted proteins typically harbors a signal peptide, consisting of approximately 15 to 30 amino acids. The SignalP prediction tool was utilized to annotate whether a protein sequence includes a signal peptide structure. Furthermore, the TMHMM tool was employed to determine the presence of transmembrane structures within a protein sequence. Ultimately, proteins that possess signal peptide structures while lacking transmembrane structures are singled out as secreted proteins. Following an analysis utilizing the SignalP 4.1 program (described in Section 2.6.), we have identified a total of 671 secreted proteins within the entirety of the genomic dataset (Table S4). Among these, the count of proteins with a prediction score surpassing 0.8 amounted to 190, constituting 28.3% of all the predicted secreted proteins. This observation signifies a higher likelihood of these proteins containing signal peptides.

3.2.5. Annotation of Putative Virulence-Associated Genes

The DFVF is a comprehensive database of fungal virulence factors, containing 2058 known pathogenic genes. Each gene is associated with details about the disease it causes and the host it targets, along with Pfam functional domain annotations and GO annotation information. By comparing the results obtained from the Diamond program (described in Section 2.6), we identified 400 homologous virulence genes, of which 38 genes (9.5%) shared more than 80% identity (Table S5). These highly homologous genes include polyubiquitin (CAA76783) in Candida albicans (100%), histone 3 (ACZ56011) in Fusarium sp. (97.7%), MAP kinase (AAS20192) in Alternaria brassicicola (95.1%), tubulin alpha-1 subunit (EAL87967) in Aspergillus fumigatus (95.1%), Ras-related protein (EAQ71072) in Pyricularia oryzae (86.7), argininosuccinate lyase (BAB40769) in Fusarium oxysporum (86.3%), Rab/GTPase (CAC41973) in Colletotrichum lindemuthianum (83.6%), and catalase (AAN04057) in Talaromyces marneffei (83.6%). These genes play critical roles in the pathogenicity of their respective species.
The Pathogen–Host Interactions (PHI) database contains verified information on virulence-associated genes that regulate pathogen host interactions. Based on the PHI analysis (described in Section 2.6), we discovered 1332 PHI putative genes in the whole genome of T. albobiverticillius, 462 of which do not affect pathogenicity (Table S6). Furthermore, 550 genes were linked to decreased virulence and 100 gene deletions entirely failed to produce illness. There were 23 increased virulence (hypervirulence) genes and 75 fatal genes, all of which were required for the pathogen’s growth, metabolism, and reproduction. Only 15 and 3 of the 1332 putative genes were effectors and chemical targets, respectively. Fusarium graminearum and Magnaporthe oryzae had the highest number of homologous genes related to virulence (118 and 117 genes, respectively), followed by Aspergillus fumigatus with 83 genes (Table 5).

3.2.6. Annotation of Secondary Metabolites Gene Clusters

Most secondary metabolites have complicated molecular structures, such as polyketides and nonribosomal peptides, which are produced by the enzymes polyketide synthase (PKS) and nonribosomal peptide synthase (NRPS), respectively. One of the crucial secondary metabolites produced by fungi is pigment, which is often produced following active cell development. Fungal pigments have reportedly been shown to have bioactive properties, which is significant for screening new drugs [13]. It can be seen from Figure 3 that the 5-day-old colony (Figure 3a) of the Tp-2 strain was cultured on a PDA plate (90 mm in diameter). As the culture time increased, the 14-day-old colony (Figure 3b) revealed the secretion of claret-red pigments into the medium. According to previous research, T. albobiverticillius was known to produce pigments that resembled azaphilones (Figure 3). FK17-P2b1, monascusone A, PP-Y, monascorubrin, PP-O, talaralbols A, and talaralbols are the names of the seven azaphilone pigments that have been isolated and reported [36]. The biosynthesis of azaphilone involves both the fatty acid synthesis route and the polyketide pathway.
There were 62 gene clusters (724 genes) involved in the secondary metabolism of T. albobiverticillius (Table S7). The gene clusters identified on the T. albobiverticillius genome encode various types of secondary metabolite biosynthetic pathways, including 18 T1PKS (type 1 PKS), 15 NRPS-like, 9 NRPS, 9 terpenes, 4 NRPS-T1PKS, 4 NRPS-like-T1PKS, 1 β-lactone, 1 NRPS-β-lactone, and 1 other (described in Section 2.6). Using the antiSMASH program, we found six PKS genes with 100% similarity in the T1PKS gene cluster. These PKS genes encode the secondary metabolites YWA1 from Aspergillus oryzae, alternariol from Parastagonospora nodorum, cichorine from Aspergillus nidulans, ACT toxin II from Alternaria alternata, and monascorubrin from Talaromyces marneffei. Upon analyzing these sequences, we discovered that the genome of T. albobiverticillius contained homologous genes that encode various secondary metabolites, such as toxins and pigments.

4. Discussion

Talaromyces now comprises 171 species, a notable increase since the genus’ reclassification in 2011, demonstrating the genus’ great diversity and widespread interest. In total, 21 species of Talaromyces have had their whole genomes sequenced as of the end of August 2023, according to the NCBI Genome Database. In this study, the genomic information of T. albobiverticillius was compared with that of four additional significant species of the Talaromyces genus. It was discovered that GC content (46%), the number of predicted genes (9523–11,447), and the number of homologous genes were virtually the same, suggesting that these markers are highly conserved throughout species. The significant variation in the genome assembly size (28.6–38.4 Mb) could be due to various sequencing techniques, depths, and assembly techniques. The comparison results of the genome phylogenetic tree revealed that T. albobiverticillius and T. atroroseus belonged to the same branch and that T. marneffei and T. verruculosus belonged to the same branch.
Remarkably, T. atroroseus, which happens to be the nearest relative to T. albobiverticillius, is renowned for its ability to produce monascus pigments, which are highly applicable in the food industry [37]. A draft genome of a prospective fungal cell factory, T. atroroseus IBT 11181 (CBS 123796), was reported in 2017 [38]. Likewise, T. verruculosus has garnered recognition for its exceptional ability to produce cellulases and antibacterial compounds [39]. In 2016, researchers unveiled the preliminary genome of T. verruculosus strain TS63-9, known for its remarkably high pectinase activity. T. marneffei is native to Southeast Asia and South China and serves as an opportunistic infectious agent in individuals with weakened immune systems [40]. The prevalence of systemic T. marneffei infection has experienced a significant surge in recent times, aligning with the rising occurrence of HIV infections [41]. T. islandicus is a globally distributed storage mold that is associated with the deterioration of grain crops following harvest, posting a threat to their quality [42]. In 2015, researchers successfully conducted whole genome sequencing and generated a preliminary version of T. islandicus, focusing on the WF-38-12 strain [43]. This particular strain exhibits a remarkably adaptable metabolism, which is evident through its ability to produce a wide variety of biopolymer-degrading enzymes, mycotoxins, and anthraquinones. These traits collectively present a diverse array of potential applications in various industrial fields. Interestingly, T. islandicus contains the most unassigned genes (515) out of all analyzed species, which is consistent with the finding that T. islandicus was farthest from the others in the phylogenetic tree. The evolutionary tree’s homology relationship might be explained by the host species or by the strain specificity within a given host species.
To predict genes, we employed various databases, including GO, KEGG, KOG, and NR. We also attempted to accurately and comprehensively annotate the gene functions of T. albobiverticillius. It was discovered that the NR database, with a total of 9782 genes, contained the most annotated genes. These findings exhibit resemblances to earlier genome-wide analyses conducted on closely related species.
T. albobiverticillius was a long-lasting latent pathogen on pomegranates, according to earlier research. On pomegranates, conidia infect epidermal cells in the stigma, hair, and silks. Along with the parenchyma and vascular cells in the silks, hyphae spread infection toward the ovary. The outcome demonstrated that T. albobiverticillius was capable of directly infecting silks and causing pulp rot through the silk channel. The fruits had no visible symptoms, although some of their pulp had turned brown before harvest. Fruits collected from afflicted orchards developed brown lesions on their peels after long-term (30–40 days) storage under ambient conditions, from which T. albobiverticillius could be isolated. To date, there has been no investigation into the pathogenic genes of T. albobiverticillius. Accordingly, the genome’s pathogenicity-related genes, such as polyubiquitin, MAP kinase, catalase, and Ras protein, were also discovered by comparing the genomes to the those in the DFVF and PHI databases. These pathogenic genes were involved in translational modification, signal transduction, and resistance to oxidative damage, which were relatively common virulence-associated factors in plant pathogenic fungi. Moreover, an abundance of gene clusters involved in secondary metabolism was found in T. albobiverticillius. The majority of these secondary metabolites were toxic polyketides, nonribosomal peptides, terpenes, and indoles that induce cell death in the host and contribute to disease development. The genomes contained many gene clusters, including T1PKS, NRPS-like, and NRPS. These secondary metabolites included alternariol, cichorine, ACT toxin II, and monascorubrin. This is consistent with previous reports that, in T. marneffei, the PKS gene PM1 is responsible for the production of yellow mitorubrin pigments (mitorubrinic acid and mitorubrinol). As a result, we suggest that T. albobiverticillius has genomic characteristics that favor virulence and pigment production.
Genome sequencing establishes a crucial molecular basis and foundation for our subsequent investigations into the gene function of T. albobiverticillius. We have successfully concluded a comprehensive study on the influence of diverse medium components, temperature variations, and culture conditions on pigment synthesis. Within this process, we also conducted a transcriptome sequencing experiment. Leveraging the fully annotated, whole-genome data, we were able to conduct a comparative analysis with the reference transcriptome data, yielding remarkably comprehensive results. Furthermore, our strategy encompasses the exploration of genes associated with pigment synthesis. We intend to target specific genes, such as the PKS1 polyketide synthases, for knockout experiments and subsequent validation of gene function. While the validation of these specific genes is pending confirmation through first-generation sequencing outcomes, the comprehensive scope of whole-genome sequencing furnishes a robust underpinning and remarkable convenience for endeavors such as gene prediction, primer design, and gene cloning.

5. Conclusions

Here, we present the genome sequence, assembly, and annotation of T. albobiverticillius. The T. albobiverticillius genome contains a rich diversity of putative genes that are also found in other plant pathogens. Furthermore, this genome sequence holds the potential to facilitate the development of molecular markers for use in evolutionary research. As the first available T. albobiverticillius genome assembly, it is poised to enhance disease management and pigment production strategies.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/jof9090909/s1, Figure S1: BUSCO assessment results of the T. albobiverticillius genome of the Tp-2 strain; Table S1: Gene ontology annotation of the T. albobiverticillius genome; Table S2: Carbohydrate-active enzyme annotation of T. albobiverticillius genome; Table S3: Transport protein annotation of T. albobiverticillius genome; Table S4: Secreted proteins annotation of T. albobiverticillius genome; Table S5: Homologous virulence genes of the T. albobiverticillius genome; Table S6: Putative genes in the PHI database of the T. albobiverticillius genome; Table S7: Secondary metabolites gene cluster annotation of the T. albobiverticillius genome.

Author Contributions

Conceptualization, T.W. and Q.N.; methodology, T.W.; software, T.W.; validation, S.C. and G.X.; formal analysis, T.W.; investigation, T.W.; resources, G.X. and C.L.; data curation, T.W.; writing—original draft preparation, T.W.; writing—review and editing, Q.N., J.Z.; visualization, T.W.; supervision, Q.N.; project administration, T.W.; funding acquisition, T.W. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the doctoral program of Nanyang Normal University Special Research Fund (No. 2022ZX039), the Science and Technique Foundation of Henan Province (No. 202102310022), and the Research Center of Henan Provincial Agricultural Biomass Resource Engineering and Technology.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The read data of Oxford Nanopore and Illumina sequencing have been submitted to the NCBI Sequence Read Archive with accession number PRJNA835810. This Whole Genome Shotgun project has been deposited at DDBJ/ENA/GenBank under the accession JAMBUR000000000. The version described in this paper is version JAMBUR020000000.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Yilmaz, N.; Visagie, C.M.; Houbraken, J.; Frisvad, J.C.; Samson, R.A. Polyphasic taxonomy of the genus Talaromyces. Stud. Mycol. 2014, 78, 175–341. [Google Scholar] [CrossRef] [PubMed]
  2. Houbraken, J.; Samson, R.A. Phylogeny of Penicillium and the segregation of Trichocomaceae into three families. Stud. Mycol. 2011, 70, 1–51. [Google Scholar] [CrossRef] [PubMed]
  3. Sun, B.-D.; Chen, A.J.; Houbraken, J.; Frisvad, J.C.; Wu, W.-P.; Wei, H.-L.; Zhou, Y.-G.; Jiang, X.-Z.; Samson, R.A. New section and species in Talaromyces. MycoKeys 2020, 68, 75–113. [Google Scholar] [CrossRef] [PubMed]
  4. Limper, A.H.; Adenis, A.; Le, T.; Harrison, T.S. Fungal infections in HIV/AIDS. Lancet Infect. Dis. 2017, 17, e334–e343. [Google Scholar] [CrossRef] [PubMed]
  5. Kumari, M.; Taritla, S.; Sharma, A.; Jayabaskaran, C. Antiproliferative and Antioxidative Bioactive Compounds in Extracts of Marine-Derived Endophytic Fungus Talaromyces purpureogenus. Front. Microbiol. 2018, 9, 1777. [Google Scholar] [CrossRef] [PubMed]
  6. Kim, M.-J.; Shim, C.-K.; Kim, Y.-K.; Hong, S.-J.; Park, J.-H.; Han, E.-J.; Kim, S.-C. Enhancement of Seed Dehiscence by Seed Treatment with Talaromyces flavus GG01 and GG04 in Ginseng (Panax ginseng). Plant Pathol. J. 2017, 33, 1–8. [Google Scholar] [CrossRef]
  7. Li, H.-L.; Li, X.-M.; Li, X.; Wang, C.-Y.; Liu, H.; Kassack, M.U.; Meng, L.-H.; Wang, B.-G. Antioxidant Hydroanthraquinones from the Marine Algal-Derived Endophytic Fungus Talaromyces islandicus EN-501. J. Nat. Prod. 2017, 80, 162–168. [Google Scholar] [CrossRef] [PubMed]
  8. Zerva, A.; Antonopoulou, I.; Enman, J.; Iancu, L.; Rova, U.; Christakopoulos, P. Cross-Linked Enzyme Aggregates of Feruloyl Esterase Preparations from Thermothelomyces thermophila and Talaromyces wortmannii. Catalysts 2018, 8, 208. [Google Scholar] [CrossRef]
  9. Varriale, S.; Houbraken, J.; Granchi, Z.; Pepe, O.; Cerullo, G.; Ventorino, V.; Chin-A-Woeng, T.; Meijer, M.; Riley, R.; Grigoriev, I.V.; et al. Talaromyces borbonicus, sp. nov., a novel fungus from biodegraded Arundo donax with potential abilities in lignocellulose conversion. Mycologia 2018, 110, 316–324. [Google Scholar] [CrossRef] [PubMed]
  10. Frisvad, J.C.; Yilmaz, N.; Thrane, U.; Rasmussen, K.B.; Houbraken, J.; Samson, R.A. Talaromyces atroroseus, a new species efficiently producing industrially relevant red pigments. PLoS ONE 2013, 8, e84102. [Google Scholar] [CrossRef] [PubMed]
  11. Venkatachalam, M.; Zelena, M.; Cacciola, F.; Ceslova, L.; Girard-Valenciennes, E.; Clerc, P.; Dugo, P.; Mondello, L.; Fouillaud, M.; Rotondo, A.; et al. Partial characterization of the pigments produced by the marine-derived fungus Talaromyces albobiverticillius 30548. Towards a new fungal red colorant for the food industry. J. Food Compost. Anal. 2018, 67, 38–47. [Google Scholar] [CrossRef]
  12. Venkatachalam, M.; Gérard, L.; Milhau, C.; Vinale, F.; Dufossé, L.; Fouillaud, M. Salinity and Temperature Influence Growth and Pigment Production in the Marine-Derived Fungal Strain Talaromyces albobiverticillius 30548. Microorganisms 2019, 7, 10. [Google Scholar] [CrossRef] [PubMed]
  13. Venkatachalam, M.; Shum-Chéong-Sing, A.; Dufossé, L.; Fouillaud, M. Statistical Optimization of the Physico-Chemical Parameters for Pigment Production in Submerged Fermentation of Talaromyces albobiverticillius 30548. Microorganisms 2020, 8, 711. [Google Scholar] [CrossRef]
  14. Mincuzzi, A.; Sanzani, S.M.; Garganese, F.; Ligorio, A.; Ippolito, A. First Report of Talaromyces albobiverticillius Causing Postharvest Fruit Rot on Pomegranate in Italy. J. Plant Pathol. 2017, 99, 303. [Google Scholar]
  15. Huang, S.; Li, T.; Yang, T.; Zheng, X.; Yang, D.; Fu, G.; Hu, G.; Tao, A.; Wang, L.; Xu, T.; et al. First Report of Pulp Rot in Externally Asymptomatic Pomegranate Fruit Caused by Talaromyces albobiverticillius in Henan Province, China. Plant Dis. 2022, 106, 1989. [Google Scholar] [CrossRef] [PubMed]
  16. Mincuzzi, A.; Sanzani, S.M.; Caputo, M.; D’Ambrosio, P.; Palou, L.; Ragni, M.; Ippolito, A. Effectiveness of alternative means for controlling pomegranate postharvest pathogens. Acta Hortic. 2023, 1363, 181–186. [Google Scholar] [CrossRef]
  17. Labuda, R.; Hudec, K.; Piecková, E.; Mezey, J.; Bohovic, R.; Mátéová, S.; Lukác, S.S. Penicillium implicatum causes a destructive rot of pomegranate fruits. Mycopathologia 2004, 157, 217–223. [Google Scholar] [CrossRef] [PubMed]
  18. Stošić, S.; Ristić, D.; Gašić, K.; Starović, M.; Ljaljević Grbić, M.; Vukojević, J.; Živković, S. Talaromyces minioluteus: New Postharvest Fungal Pathogen in Serbia. Plant Dis. 2020, 104, 656–667. [Google Scholar] [CrossRef] [PubMed]
  19. El-Ashram, S.; Al Nasr, I.; Suo, X. Nucleic acid protocols: Extraction and optimization. Biotechnol. Rep. 2016, 12, 33–39. [Google Scholar] [CrossRef] [PubMed]
  20. Emms, D.M.; Kelly, S. OrthoFinder: Phylogenetic orthology inference for comparative genomics. Genome Biol. 2019, 20, 238. [Google Scholar] [CrossRef]
  21. Minh, B.Q.; Schmidt, H.A.; Chernomor, O.; Schrempf, D.; Woodhams, M.D.; von Haeseler, A.; Lanfear, R. IQ-TREE 2: New Models and Efficient Methods for Phylogenetic Inference in the Genomic Era. Mol. Biol. Evol. 2020, 37, 1530–1534. [Google Scholar] [CrossRef]
  22. Saha, S.; Bridges, S.; Magbanua, Z.V.; Peterson, D.G. Empirical comparison of ab initio repeat finding programs. Nucleic Acids Res. 2008, 36, 2284–2294. [Google Scholar] [CrossRef]
  23. Benson, G. Tandem repeats finder: A program to analyze DNA sequences. Nucleic Acids Res. 1999, 27, 573–580. [Google Scholar] [CrossRef]
  24. Lowe, T.M.; Eddy, S.R. tRNAscan-SE: A program for improved detection of transfer RNA genes in genomic sequence. Nucleic Acids Res. 1997, 25, 955–964. [Google Scholar] [CrossRef]
  25. Lagesen, K.; Hallin, P.; Rødland, E.A.; Staerfeldt, H.-H.; Rognes, T.; Ussery, D.W. RNAmmer: Consistent and rapid annotation of ribosomal RNA genes. Nucleic Acids Res. 2007, 35, 3100–3108. [Google Scholar] [CrossRef]
  26. Gardner, P.P.; Daub, J.; Tate, J.G.; Nawrocki, E.P.; Kolbe, D.L.; Lindgreen, S.; Wilkinson, A.C.; Finn, R.D.; Griffiths-Jones, S.; Eddy, S.R.; et al. Rfam: Updates to the RNA families database. Nucleic Acids Res. 2009, 37, D136–D140. [Google Scholar] [CrossRef]
  27. Ashburner, M.; Ball, C.A.; Blake, J.A.; Botstein, D.; Butler, H.; Cherry, J.M.; Davis, A.P.; Dolinski, K.; Dwight, S.S.; Eppig, J.T.; et al. Gene ontology: Tool for the unification of biology. The Gene Ontology Consortium. Nat. Genet. 2000, 25, 25–29. [Google Scholar] [CrossRef]
  28. Kanehisa, M.; Goto, S.; Hattori, M.; Aoki-Kinoshita, K.F.; Itoh, M.; Kawashima, S.; Katayama, T.; Araki, M.; Hirakawa, M. From genomics to chemical genomics: New developments in KEGG. Nucleic Acids Res. 2006, 34, D354–D357. [Google Scholar] [CrossRef]
  29. Galperin, M.Y.; Makarova, K.S.; Wolf, Y.I.; Koonin, E.V. Expanded microbial genome coverage and improved protein family annotation in the COG database. Nucleic Acids Res. 2015, 43, D261–D269. [Google Scholar] [CrossRef] [PubMed]
  30. Saier, M.H.; Reddy, V.S.; Tsu, B.V.; Ahmed, M.S.; Li, C.; Moreno-Hagelsieb, G. The Transporter Classification Database (TCDB): Recent advances. Nucleic Acids Res. 2016, 44, D372–D379. [Google Scholar] [CrossRef]
  31. Bairoch, A.; Apweiler, R. The SWISS-PROT protein sequence database and its supplement TrEMBL in 2000. Nucleic Acids Res. 2000, 28, 45–48. [Google Scholar] [CrossRef] [PubMed]
  32. Petersen, T.N.; Brunak, S.; von Heijne, G.; Nielsen, H. SignalP 4.0: Discriminating signal peptides from transmembrane regions. Nat. Methods 2011, 8, 785–786. [Google Scholar] [CrossRef] [PubMed]
  33. Medema, M.H.; Blin, K.; Cimermancic, P.; de Jager, V.; Zakrzewski, P.; Fischbach, M.A.; Weber, T.; Takano, E.; Breitling, R. antiSMASH: Rapid identification, annotation and analysis of secondary metabolite biosynthesis gene clusters in bacterial and fungal genome sequences. Nucleic Acids Res. 2011, 39, W339–W346. [Google Scholar] [CrossRef]
  34. Urban, M.; Pant, R.; Raghunath, A.; Irvine, A.G.; Pedro, H.; Hammond-Kosack, K.E. The Pathogen-Host Interactions database (PHI-base): Additions and future developments. Nucleic Acids Res. 2015, 43, D645–D655. [Google Scholar] [CrossRef]
  35. Cantarel, B.L.; Coutinho, P.M.; Rancurel, C.; Bernard, T.; Lombard, V.; Henrissat, B. The Carbohydrate-Active EnZymes database (CAZy): An expert resource for Glycogenomics. Nucleic Acids Res. 2009, 37, D233–D238. [Google Scholar] [CrossRef] [PubMed]
  36. Bai, W.; Jing, L.-L.; Guan, Q.-Y.; Tan, R.-X. Two new azaphilone pigments from Talaromyces albobiverticillius and their anti-inflammatory activity. J. Asian Nat. Prod. Res. 2021, 23, 325–332. [Google Scholar] [CrossRef]
  37. Ruiz-Sánchez, J.P.; Morales-Oyervides, L.; Giuffrida, D.; Dufossé, L.; Montañez, J.C. Production of Pigments under Submerged Culture through Repeated Batch Fermentation of Immobilized Talaromyces atroroseus GH2. Fermentation 2023, 9, 171. [Google Scholar] [CrossRef]
  38. Thrane, U.; Rasmussen, K.B.; Petersen, B.; Rasmussen, S.; Sicheritz-Pontén, T.; Mortensen, U.H. Genome Sequence of Talaromyces atroroseus, Which Produces Red Colorants for the Food Industry. Genome Announc. 2017, 5, e01736-16. [Google Scholar] [CrossRef]
  39. Chadni, Z.; Rahaman, M.H.; Jerin, I.; Hoque, K.; Reza, M.A. Extraction and optimisation of red pigment production as secondary metabolites from Talaromyces verruculosus and its potential use in textile industries. Mycology 2017, 8, 48–57. [Google Scholar] [CrossRef]
  40. He, L.; Mei, X.; Lu, S.; Ma, J.; Hu, Y.; Mo, D.; Chen, X.; Fan, R.; Xi, L.; Xie, T. Talaromyces marneffei infection in non-HIV-infected patients in mainland China. Mycoses 2021, 64, 1170–1176. [Google Scholar] [CrossRef]
  41. Ning, C.; Lai, J.; Wei, W.; Zhou, B.; Huang, J.; Jiang, J.; Liang, B.; Liao, Y.; Zang, N.; Cao, C.; et al. Accuracy of rapid diagnosis of Talaromyces marneffei: A systematic review and meta-analysis. PLoS ONE 2018, 13, e0195569. [Google Scholar] [CrossRef] [PubMed]
  42. Fukaya, M.; Ozaki, T.; Minami, A.; Oikawa, H. Biosynthetic machineries of anthraquinones and bisanthraquinones in Talaromyces islandicus. Biosci. Biotechnol. Biochem. 2022, 86, 435–443. [Google Scholar] [CrossRef] [PubMed]
  43. Schafhauser, T.; Wibberg, D.; Rückert, C.; Winkler, A.; Flor, L.; van Pée, K.-H.; Fewer, D.P.; Sivonen, K.; Jahn, L.; Ludwig-Müller, J.; et al. Draft genome sequence of Talaromyces islandicus (“Penicillium islandicum”) WF-38-12, a neglected mold with significant biotechnological potential. J. Biotechnol. 2015, 211, 101–102. [Google Scholar] [CrossRef] [PubMed]
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Figure 1. Genomic landscape of Talaromyces albobiverticillius strain Tp-2 visualized using Circos software (version 0.69-9). The outermost circle represents the location coordinates of the genome sequence. Moving inward, the subsequent circles depict GC content, GC-skew, density of coding genes, rRNA, snRNA, and tRNA. The curves connecting the different regions in the central circle represent gene duplications.
Figure 1. Genomic landscape of Talaromyces albobiverticillius strain Tp-2 visualized using Circos software (version 0.69-9). The outermost circle represents the location coordinates of the genome sequence. Moving inward, the subsequent circles depict GC content, GC-skew, density of coding genes, rRNA, snRNA, and tRNA. The curves connecting the different regions in the central circle represent gene duplications.
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Figure 2. A phylogenetic tree of five Talaromyces species constructed based on whole genome sequence data using IQ-Tree.
Figure 2. A phylogenetic tree of five Talaromyces species constructed based on whole genome sequence data using IQ-Tree.
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Figure 3. Colonies and conidia morphology of the Tp-2 strain under an electron microscope. (a) A 5-day-old colony of the Tp-2 strain cultivated on a PDA plate (90 mm in diameter) at 28 °C. (b) A 14-day-old colony, revealing the secretion of claret-red pigments into the medium. (c) Conidia and strigmata visualized under a scanning electron microscope. Scale bar = 10 μm.
Figure 3. Colonies and conidia morphology of the Tp-2 strain under an electron microscope. (a) A 5-day-old colony of the Tp-2 strain cultivated on a PDA plate (90 mm in diameter) at 28 °C. (b) A 14-day-old colony, revealing the secretion of claret-red pigments into the medium. (c) Conidia and strigmata visualized under a scanning electron microscope. Scale bar = 10 μm.
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Table 1. Genome assembly statistics of Talaromyces albobiverticillius strain Tp-2.
Table 1. Genome assembly statistics of Talaromyces albobiverticillius strain Tp-2.
VariablesStatistics
Genome assembly size (bp)38,354,882
Number of contigs14
Contigs N50 (bp) 4,594,200
Maximum contigs length (bp) 6,575,826
GC content (%) 45.78
BUSCO completeness (%) * 98.3
Fragmented BUSCOs (%)0.3
Missing BUSCOs (%)1.4
Total BUSCOs searched758
SINEs (bp)2322
LINEs (bp)32,971
LTR (bp)175,050
DNA transposons (bp)116,497
Total length of Repeats (bp)321,868
TR (bp)244,896
Minisatellite DNA (bp)161,135
Microsatellite DNA (bp)18,152
Protein-coding genes10,380
Mean gene length (bp)1432
* BUSCO, Benchmarking universal single-copy orthologs; SINEs: Short interspersed nuclear elements; LINEs: Long interspersed nuclear elements; LTR: Long terminal repeat retrotransposons; TR: Tandem repeat.
Table 2. Genome assembly statistics of Talaromyces albobiverticillius and phylogenetically related Talaromyces taxa.
Table 2. Genome assembly statistics of Talaromyces albobiverticillius and phylogenetically related Talaromyces taxa.
Genome
Statistics		Talaromyces
albobiverticillius		Talaromyces
atroroseus		Talaromyces
islandicus		Talaromyces
marneffei		Talaromyces
verruculosus
strainTp-2IBT 11181WF-38-12ATCC 18224TS63-9
hostPunica granatumCapsicum annuumTriticum aestivumHomo sapiensNicotiana tabacum
countryChinaDenmarkGermanyUSAChina
sequencing
technology		Oxford
Nanopore		Illumina HiSeqIllumina MiSeqSOLiDIllumina HiSeq
assembly size (Mb)38.430.834.728.637.6
protein-coding10,3809523992710,02311,447
GC content (%)4646464646
unassigned
genes 1		381325515429398
one-to-one
orthologous 2		—6396580958716493
1 Unassigned genes were not assigned to any orthogroups of these five species genome data. 2 Orthologous genes compared to T. albobiverticillius genes.
Table 3. Gene functional annotation of Talaromyces albobiverticillius strain Tp-2.
Table 3. Gene functional annotation of Talaromyces albobiverticillius strain Tp-2.
VariablesStatistics
Protein-coding genes10,380
Genes annotated by Pfam7412
Genes annotated by KEGG8844
Genes annotated by KOG2160
Genes annotated by NR9782
Genes annotated by Swiss-Prot3657
Type I polyketide synthases genes214
Cytochrome P450 genes227
Virulence genes 400
Carbohydrate active enzymes 750
Transmembrane proteins 2057
Transport proteins638
Secreted Protein671
Table 4. Categories of carbohydrate-active enzymes in Talaromyces albobiverticillius strain Tp-2.
Table 4. Categories of carbohydrate-active enzymes in Talaromyces albobiverticillius strain Tp-2.
CategoriesFamiliesGenes
Glycoside Hydrolases (GHs)105427
Glycosyltransferases (GTs)37121
Polysaccharide Lyases (PLs)811
Carbohydrate Esterases (CEs)1040
Auxiliary Activities (AAs)1460
Carbohydrate-Binding Modules (CBMs)1291
Table 5. Putative genes of the Pathogen–Host Interactions database in Talaromyces albobiverticillius strain Tp-2.
Table 5. Putative genes of the Pathogen–Host Interactions database in Talaromyces albobiverticillius strain Tp-2.
Pathogen SpeciesPHI DatabaseAssociated with Virulence 1
Alternaria alternata108
Aspergillus flavus1515
Aspergillus fumigatus14783
Beauveria bassiana4530
Bipolaris maydis1111
Botrytis cinerea4118
Candida albicans4435
Colletotrichum gloeosporioides108
Colletotrichum graminicola76
Cryptococcus neoformans4029
Fusarium graminearum366118
Magnaporthe oryzae163117
Sclerotinia sclerotiorum1711
Verticillium dahliae1911
Zymoseptoria tritici178
1 Genes associated with virulence, including loss, reduced, and increased virulence genes, chemistry targets, and effectors (plant virulence determinants).
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Wang, T.; Chen, S.; Niu, Q.; Xu, G.; Lu, C.; Zhang, J. Genomic Sequence Resource of Talaromyces albobiverticillius, the Causative Pathogen of Pomegranate Pulp Rot Disease. J. Fungi 2023, 9, 909. https://doi.org/10.3390/jof9090909

AMA Style

Wang T, Chen S, Niu Q, Xu G, Lu C, Zhang J. Genomic Sequence Resource of Talaromyces albobiverticillius, the Causative Pathogen of Pomegranate Pulp Rot Disease. Journal of Fungi. 2023; 9(9):909. https://doi.org/10.3390/jof9090909

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Wang, Tan, Shuchang Chen, Qiuhong Niu, Guangling Xu, Chenxu Lu, and Jin Zhang. 2023. "Genomic Sequence Resource of Talaromyces albobiverticillius, the Causative Pathogen of Pomegranate Pulp Rot Disease" Journal of Fungi 9, no. 9: 909. https://doi.org/10.3390/jof9090909

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