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基于差异表达结合单核苷酸多态性关联分析和连锁图谱,筛选和验证马铃薯对致病疫霉成熟校正抗性的候选基因

Selection and validation of potato candidate genes for maturity corrected resistance to Phytophthora infestans based on differential expression combined with SNP association and linkage mapping.

作者信息

Muktar Meki S, Lübeck Jens, Strahwald Josef, Gebhardt Christiane

机构信息

Department for Plant Breeding and Genetics, Max Planck Institute for Plant Breeding Research Cologne, Germany.

Saka-Pflanzenzucht GmbH & Co. KG Windeby, Germany.

出版信息

Front Genet. 2015 Sep 23;6:294. doi: 10.3389/fgene.2015.00294. eCollection 2015.

Abstract

Late blight of potato (Solanum tuberosum L.) caused by the oomycete Phytophthora infestans (Mont.) de Bary, is one of the most important bottlenecks of potato production worldwide. Cultivars with high levels of durable, race unspecific, quantitative resistance are part of a solution to this problem. However, breeding for quantitative resistance is hampered by the correlation between resistance and late plant maturity, which is an undesirable agricultural attribute. The objectives of our research are (i) the identification of genes that condition quantitative resistance to P. infestans not compromised by late plant maturity and (ii) the discovery of diagnostic single nucleotide polymorphism (SNP) markers to be used as molecular tools to increase efficiency and precision of resistance breeding. Twenty two novel candidate genes were selected based on comparative transcript profiling by SuperSAGE (serial analysis of gene expression) in groups of plants with contrasting levels of maturity corrected resistance (MCR). Reproducibility of differential expression was tested by quantitative real time PCR and allele specific pyrosequencing in four new sets of genotype pools with contrasting late blight resistance levels, at three infection time points and in three independent infection experiments. Reproducibility of expression patterns ranged from 28 to 97%. Association mapping in a panel of 184 tetraploid cultivars identified SNPs in five candidate genes that were associated with MCR. These SNPs can be used in marker-assisted resistance breeding. Linkage mapping in two half-sib families (n = 111) identified SNPs in three candidate genes that were linked with MCR. The differentially expressed genes that showed association and/or linkage with MCR putatively function in phytosterol synthesis, fatty acid synthesis, asparagine synthesis, chlorophyll synthesis, cell wall modification, and in the response to pathogen elicitors.

摘要

由卵菌致病疫霉(Phytophthora infestans (Mont.) de Bary)引起的马铃薯晚疫病是全球马铃薯生产中最重要的制约因素之一。具有高水平持久、小种非特异性、数量抗性的品种是解决这一问题的部分方案。然而,数量抗性育种受到抗性与植株晚熟之间相关性的阻碍,而晚熟是一种不理想的农艺性状。我们研究的目标是:(i)鉴定决定对致病疫霉数量抗性且不受植株晚熟影响的基因;(ii)发现诊断性单核苷酸多态性(SNP)标记,用作分子工具以提高抗性育种的效率和精度。基于在成熟校正抗性(MCR)水平不同的植株组中通过SuperSAGE(基因表达序列分析)进行的比较转录谱分析选择了22个新的候选基因。通过定量实时PCR和等位基因特异性焦磷酸测序,在具有不同晚疫病抗性水平的四组新基因型库中、在三个感染时间点以及在三个独立的感染实验中,测试了差异表达的可重复性。表达模式的可重复性范围为28%至97%。在一组184个四倍体品种中进行关联作图,鉴定出五个与MCR相关的候选基因中的SNP。这些SNP可用于标记辅助抗性育种。在两个半同胞家系(n = 111)中进行连锁作图,鉴定出三个与MCR连锁的候选基因中的SNP。与MCR显示关联和/或连锁的差异表达基因可能在植物甾醇合成、脂肪酸合成、天冬酰胺合成、叶绿素合成、细胞壁修饰以及对病原体激发子的反应中发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2452/4585299/ca6dfb59688a/fgene-06-00294-g0001.jpg

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