Heilongjiang Academy of Agricultural Sciences, Harbin, China.
College of Agriculture, Northeast Agriculture University, Harbin, China.
Sci Rep. 2020 Oct 22;10(1):18065. doi: 10.1038/s41598-020-75235-x.
Outbreaks of gummy stem blight (GSB), an emerging seed pumpkin disease, have increased in number and have become more widespread in recent years. Previously we reported that Stagonosporopsis cucurbitacearum (Sc.) is the dominant fungal cause of pumpkin seedling GSB in Northeast China, where it has greatly reduced crop yields in that region. Here, high-throughput whole-genome sequencing and assembly of the Sc. genome were conducted toward revealing pathogenic molecular regulatory mechanisms involved in fungal growth and development. Zq-1 as representative Sc. strain, DNA of Zq-1was prepared for genomic sequencing, we obtained 5.24 Gb of high-quality genomic sequence data via PacBio RS II sequencing. After sequence data was processed to filter out low quality reads, a hierarchical genome-assembly process was employed that generated a genome sequence of 35.28 Mb in size. A total of 9844 genes were predicted, including 237 non-coding RNAs, 1024 genes encoding proteins with signal peptides, 2066 transmembrane proteins and 756 secretory proteins.Transcriptional identification revealed 54 differentially expressed secretory proteins. Concurrently, 605, 130 and 2869 proteins were matched in the proprietary databases Carbohydrate-Active EnZymes database (CAZyme), Transporter Classification Database (TCDB) and Pathogen-Host Interactions database (PHI), respectively. And 96 and 36 DEGs were identified form PHI database and CAZyme database, respectively. In addition, contig00011.93 was an up-regulated DEG involving ATP-binding cassette metabolism in the procession of infection. In order to test relevance of gene predictions to GSB, DEGs with potential pathogenic relevance were revealed through transcriptome data analysis of Sc. strains pre- and post-infection of pumpkin. Interestingly, Sc. and Leptosphaeria maculans (Lm.) exhibited relatively similar with genome lengths, numbers of protein-coding genes and other characteristics. This work provides a foundation for future exploration of additional Sc. gene functions toward the development of more effective GSB control strategies.
近年来,一种新兴的瓜类种实疫病——胶质茎溃疡病(GSB)的爆发次数有所增加,且范围也更加广泛。此前我们曾报道,链格孢(Stagonosporopsis cucurbitacearum (Sc.))是东北地区南瓜种实 GSB 的主要真菌病原体,这极大地降低了该地区的作物产量。在这里,我们进行了高通量全基因组测序和 Sc. 基因组组装,以揭示参与真菌生长和发育的致病分子调控机制。以 Zq-1 作为代表性的 Sc. 菌株,我们对其 DNA 进行了基因组测序准备,通过 PacBio RS II 测序获得了 5.24Gb 的高质量基因组序列数据。在对序列数据进行处理以过滤低质量读数之后,采用了分层基因组组装流程,生成了大小为 35.28Mb 的基因组序列。共预测到 9844 个基因,包括 237 个非编码 RNA、1024 个编码具有信号肽的蛋白质的基因、2066 个跨膜蛋白和 756 个分泌蛋白。转录鉴定揭示了 54 个差异表达的分泌蛋白。同时,在碳水化合物活性酶数据库(CAZyme)、转运蛋白分类数据库(TCDB)和病原体-宿主相互作用数据库(PHI)中分别匹配了 605、130 和 2869 个蛋白质。并且从 PHI 数据库和 CAZyme 数据库中分别鉴定到 96 个和 36 个 DEG。此外,在感染过程中,contig00011.93 是一个上调的 DEG,涉及 ATP 结合盒代谢。为了测试基因预测与 GSB 的相关性,通过 Sc. 菌株在感染前后的转录组数据分析,揭示了具有潜在致病相关性的 DEG。有趣的是,Sc. 和麦角硫因(Lm.)的基因组长度、蛋白质编码基因数量和其他特征相对相似。这项工作为进一步探索 Sc. 基因功能提供了基础,以开发更有效的 GSB 控制策略。