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在马铃薯选育系 SW93-1015 中,利用效应子驱动的标记开发和抗疫霉属(Phytophthora infestans)抗性基因的克隆。

Effector-driven marker development and cloning of resistance genes against Phytophthora infestans in potato breeding clone SW93-1015.

机构信息

Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Alnarp, Sweden.

Department of Plant Breeding, Swedish University of Agricultural Sciences, Alnarp, Sweden.

出版信息

Theor Appl Genet. 2016 Jan;129(1):105-15. doi: 10.1007/s00122-015-2613-y. Epub 2015 Oct 30.

Abstract

We show the usefulness of integrating effector screening in a breeding program and in resistance gene cloning, with Phytophthora resistance in the Swedish potato breeding clone SW93-1015 as an example. Phytophthora infestans is one of the most devastating plant pathogens worldwide. We have earlier found that the SW93-1015 potato breeding clone has an efficient resistance against P. infestans under field conditions in Sweden, which has an unusually high local diversity of the pathogen. This potato clone has characteristics that are different from classical R-gene-mediated resistance such as elevated levels of hydrogen peroxide (H2O2) under controlled conditions. Analysis of 76 F1 potato progenies from two individual crosses resulted in nearly 50% resistant clones, from both crosses. This result suggests that the SW93-1015 clone has a simplex genotype for this trait. Screening with over 50 different P. infestans effectors, containing the conserved motif RXLR (for Arg, any amino acid, Leu, Arg), revealed a specific response to Avr2, which suggests that SW93-1015 might contain a functional homolog of the R2 resistance gene. We cloned eight R2 gene homologs from SW93-1015, whereof seven have not been described before and one gene encoded a protein identical to Rpi-ABPT. Expression of this gene in potato cultivar Désirée provided R2-specific resistance, whereas other homologues did not. Using RNAseq analyses we designed a new DNA marker for the R2 resistance in SW93-1015. In summary, we have demonstrated the use of effector screening in practical breeding material and revealed the key resistance mechanism for SW93-1015.

摘要

我们展示了在一个育种计划和抗性基因克隆中整合效应子筛选的有用性,以瑞典马铃薯品种 SW93-1015 中的 Phytophthora 抗性为例。Phytophthora infestans 是全球最具破坏性的植物病原体之一。我们之前发现,SW93-1015 马铃薯品种在瑞典田间条件下对 P. infestans 具有高效抗性,而瑞典的病原体具有异常高的本地多样性。该马铃薯品种具有不同于经典 R 基因介导抗性的特征,例如在受控条件下过氧化氢 (H2O2) 水平升高。对来自两个个体杂交的 76 个 F1 马铃薯后代的分析导致近 50%的抗性克隆来自两个杂交种。这一结果表明,SW93-1015 克隆对此性状具有单倍型基因型。用超过 50 种不同的 P. infestans 效应子进行筛选,其中包含保守基序 RXLR(用于 Arg、任何氨基酸、Leu、Arg),发现对 Avr2 有特异性反应,这表明 SW93-1015 可能含有 R2 抗性基因的功能同源物。我们从 SW93-1015 中克隆了 8 个 R2 基因同源物,其中 7 个以前没有描述过,一个基因编码的蛋白质与 Rpi-ABPT 相同。在马铃薯品种 Désirée 中表达该基因提供了 R2 特异性抗性,而其他同源物则没有。使用 RNAseq 分析,我们为 SW93-1015 中的 R2 抗性设计了一个新的 DNA 标记。总之,我们已经证明了在实际育种材料中使用效应子筛选的有效性,并揭示了 SW93-1015 的关键抗性机制。

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