基于基因混池测序技术的小麦条锈菌对三唑酮抗性位点相关分析

Identification of triadimefon resistance loci in wheat stripe rust fungus based on gene pool sequencing

为探究小麦条锈菌Puccinia striiformis f. sp. tritici对三唑类杀菌剂产生抗药性的相关机理,以小麦条锈菌抗药性菌株YQ324与敏感性菌株贵1-2在转主寄主堆花小檗Berberis aggregata上构建的有性遗传群体为材料,通过离体叶段法对F1代杂交及F2代自交后代的夏孢子群体进行三唑酮敏感性生物学测定,并在F2代夏孢子群体中筛选20株抗药性菌株和20株敏感性菌株分别构建DNA抗、感池。在对抗、感混池进行高通量测序之后,通过质量检测和数据分析得到2个极端混池中的变异信息。根据ΔIndex为1的筛选标准在小麦条锈菌中鉴定出67个与三唑酮抗性显著相关的单核苷酸多态性(single nucleotide polymorphism,SNP)位点;利用SNPEff软件对这些SNP位点引起氨基酸非同义突变所在的基因序列进行功能注释,最终在pcontig030上筛选到Pst104E09436和Pst104E09437这2个基因作为小麦条锈菌对三唑酮抗性相关的候选基因。

In order to explore the mechanism of resistance in Puccinia striiformis f. sp. tritici, the wheat stripe rust fungus, to triadimefon, the resistant strain YQ324 and sensitive strain Gui 1-2 of the fungus were hybridized on Berberis aggregata and a F2 uredinial population was constructed. After evaluating the triadimefon resistance for the F1 hybridizing and F2 selfing uredinial populations using a detached leaf method in vitro, the resistant and susceptible DNA bulks were prepared, both of which included 20 extreme individuals. After high-throught put sequencing, the genomic variant information in the two bulks was obtained through quality detection control and data analysis. Using a screening criterion of Δindex equal to one, 67 SNP loci of wheat stripe rust fungus were identified. SNPEff software was used to annotate the gene sequence that contained at least one significant SNP locus causing a non-synonymous mutation in amino acid. Finally, Pst104 E09436 and Pst104 E09437 genes were identified on pcontig030 as candidate genes for triadimefon resistance in wheat stripe rust fungus.