RenSeq on Sr27 carrying triticale variety Coorong and 4 susceptible EMS mutants

Seeds of triticale variety Coorong carrying stem rust resistance (R) gene Sr27 were subjected to EMS mutagenesis. Screening of M2 families with wheat stem rust pathogen Pgt21-0 inoculation identified several susceptible mutants. Genomic DNA from wildtype and 4 susceptible mutants were enriched using a bait library specific for Triticeae NB-LRRs (V2). This NLR gene capture was performed by Arbor Biosciences (Ann Arbor, USA) following the myBaits protocol. Enriched libraries were sequenced by HiSeq 2500 (Illumina) to produce 250 bp paired-end reads.

携带茎锈病抗性（R）基因Sr27的小黑麦品种库伦（Coorong）的种子经乙基甲磺酸（EMS）诱变处理。随后采用小麦茎锈病病原菌Pgt21-0接种筛选M2代株系，成功鉴定出多个感病突变体。提取野生型材料与4株感病突变体的基因组DNA，使用针对小麦族NB-LRR（核苷酸结合位点-富亮氨酸重复序列，Triticeae NB-LRRs）的V2版特异性捕获探针库进行富集。该NLR基因捕获实验由美国安阿伯的Arbor Biosciences公司依照myBaits操作流程完成。富集后的文库通过Illumina公司的HiSeq 2500测序平台进行测序，生成250 bp双端测序读段。