Prasanth C Naveen, Viswanathan R, Malathi P, Sundar A Ramesh
Division of Crop Protection, Sugarcane Breeding Institute, Indian Council of Agricultural Research, Tamil Nadu, Coimbatore, 641007 India.
3 Biotech. 2021 Jan;11(1):20. doi: 10.1007/s13205-020-02572-z. Epub 2021 Jan 3.
an intriguing pathogen causing red rot in sugarcane, exhibits enormous variation for pathogenicity under field conditions. A species-specific marker is very much needed to classify the virulence among the varying population and to identify the potential of a pathotype by mining the microsatellites, which are considered to be the largest genetic source to develop molecular markers for an organism. In this study, we have mined the genome using MISA database which yielded 12,121 SSRs from 48.1 Mb and 2745 SSRs containing sequences. The most frequent SSR types from the genome of was di-nucleotide which constitutes 50.89% followed by tri-nucleotide 39.60%, hepta-nucleotide 6.7%, hexa-nucleotide 1.38% and penta-nucleotide 1.3%. Over 90 SSR containing sequences from the genome were predicted using BlastX which are found to be non-homologs. Most of the annotated SSR containing sequences fell in CAZy superfamilies, proteases, peptidases, plant cell wall degrading enzymes (PCDWE) and membrane transporters which are considered to be pathogenicity gene clusters. Among them, glycosyl hydrolases (GH) were found to be abundant in SSR containing sequences which again proved our previous transcriptome results. Our results suggested that the mined microsatellites from genome show absence of homolog sequences which suggests that these markers could be used as an ideal species-specific molecular marker. Two virulence specific markers were characterized using conventional PCR assays from along with virulent species-specific (VSS) marker developed for . The study lays the foundation for the development of specific molecular marker to phenotype the pathotypes based on virulence.
一种引起甘蔗赤腐病的有趣病原体,在田间条件下致病性表现出巨大差异。非常需要一种物种特异性标记来对不同群体中的毒力进行分类,并通过挖掘微卫星来鉴定致病型的潜力,微卫星被认为是为生物体开发分子标记的最大遗传来源。在本研究中,我们使用MISA数据库挖掘了基因组,该数据库从48.1 Mb中产生了12,121个SSR和2745个含SSR的序列。该基因组中最常见的SSR类型是二核苷酸,占50.89%,其次是三核苷酸,占39.60%,七核苷酸占6.7%,六核苷酸占1.38%,五核苷酸占1.3%。使用BlastX预测了该基因组中超过90个含SSR的序列,发现它们是无同源物的。大多数注释的含SSR序列属于碳水化合物活性酶(CAZy)超家族、蛋白酶、肽酶、植物细胞壁降解酶(PCDWE)和膜转运蛋白,这些被认为是致病基因簇。其中,糖基水解酶(GH)在含SSR的序列中含量丰富,这再次证明了我们之前的转录组结果。我们的结果表明,从该基因组中挖掘的微卫星显示没有同源序列,这表明这些标记可作为理想的物种特异性分子标记。使用常规PCR分析从该病原体中鉴定了两个毒力特异性标记,以及为另一种病原体开发的毒力物种特异性(VSS)标记。该研究为开发基于毒力对致病型进行表型分析的该病原体特异性分子标记奠定了基础。