Dissection of a rapidly evolving wheat resistance gene cluster by long-read genome sequencing accelerated the cloning of Pm69
2023; Elsevier BV; Volume: 5; Issue: 1 Linguagem: Inglês
10.1016/j.xplc.2023.100646
ISSN2590-3462
AutoresYinghui Li, Zhen-Zhen Wei, Hanan Sela, Liubov Govta, Valentyna Klymiuk, Rajib Roychowdhury, Harmeet Singh Chawla, Jennifer Ens, Krystalee Wiebe, Valeria Bocharova, Roi Ben‐David, Prerna B. Pawar, Yuqi Zhang, Samidha Jaiwar, István Molnár, Jaroslav Doležel, Gitta Coaker, Curtis Pozniak, Tzion Fahima,
Tópico(s)Plant Virus Research Studies
ResumoGene cloning in repeat-rich polyploid genomes remains challenging. Here we describe a strategy for overcoming major bottlenecks in the cloning of the powdery mildew (Pm) resistance gene (R-gene) Pm69 derived from tetraploid wild emmer wheat (WEW). A conventional positional cloning approach was not effective due to suppressed recombination. Chromosome sorting was compromised by insufficient purity. A Pm69 physical map, constructed by assembling Oxford Nanopore Technology (ONT) long-read genome sequences, revealed a rapidly evolving nucleotide-binding leucine-rich repeat (NLR) R-gene cluster with structural variations. A single candidate NLR was identified by anchoring RNASeq reads of susceptible mutants to ONT contigs and was validated by virus-induced gene silencing. Pm69 is likely a newly evolved NLR, which was discovered only in one location across the WEW distribution range in Israel. Pm69 was successfully introgressed into cultivated wheat, and a diagnostic molecular marker was used to accelerate its deployment and pyramiding with other R-genes.
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