Zhao Chuanzhi, Qiu Jingjing, Agarwal Gaurav, Wang Jiangshan, Ren Xuezhen, Xia Han, Guo Baozhu, Ma Changle, Wan Shubo, Bertioli David J, Varshney Rajeev K, Pandey Manish K, Wang Xingjun
Biotechnology Research Center, Shandong Academy of Agricultural Sciences and Shandong Provincial Key Laboratory of Crop Genetic Improvement, Ecology and PhysiologyJinan, China.
College of Life Sciences, Shandong Normal UniversityJinan, China.
Front Plant Sci. 2017 Jul 18;8:1209. doi: 10.3389/fpls.2017.01209. eCollection 2017.
Despite several efforts in the last decade toward development of simple sequence repeat (SSR) markers in peanut, there is still a need for more markers for conducting different genetic and breeding studies. With the effort of the International Peanut Genome Initiative, the availability of reference genome for both the diploid progenitors of cultivated peanut allowed us to identify 135,529 and 199,957 SSRs from the A () and B genomes (), respectively. Genome sequence analysis showed uneven distribution of the SSR motifs across genomes with variation in parameters such as SSR type, repeat number, and SSR length. Using the flanking sequences of identified SSRs, primers were designed for 51,354 and 60,893 SSRs with densities of 49 and 45 SSRs per Mb in and , respectively. PCR analysis of these SSR markers showed high transferability between wild and cultivated species. Two physical maps were developed for the A genome and the B genome using these SSR markers, and two reported disease resistance quantitative trait loci (QTLs), for tomato spotted wilt virus (TSWV) and for leaf spot (LS), were mapped in the 8.135 Mb region of chromosome A04 of . From this genomic region, 719 novel SSR markers were developed, which provide the possibility for fine mapping of these QTLs. In addition, this region also harbors 652 genes and 49 of these are defense related genes, including two NB-ARC genes, three LRR receptor-like genes and three WRKY transcription factors. These disease resistance related genes could contribute to resistance to viral (such as TSWV) and fungal (such as LS) diseases in peanut. In summary, this study not only provides a large number of molecular markers for potential use in peanut genetic map development and QTL mapping but also for map-based gene cloning and molecular breeding.
尽管在过去十年里人们为开发花生的简单序列重复(SSR)标记付出了诸多努力,但开展不同的遗传和育种研究仍需要更多标记。在国际花生基因组计划的努力下,栽培花生二倍体祖先的参考基因组的可得性使我们能够分别从A()基因组和B基因组()中鉴定出135,529个和199,957个SSR。基因组序列分析表明,SSR基序在基因组中的分布不均匀,在SSR类型、重复次数和SSR长度等参数上存在差异。利用鉴定出的SSR的侧翼序列,分别为51,354个和60,893个SSR设计了引物,在和中SSR密度分别为每兆碱基49个和45个。对这些SSR标记进行的PCR分析表明,它们在野生和栽培物种之间具有很高的可转移性。利用这些SSR标记为A基因组和B基因组构建了两个物理图谱,并且在的A04染色体的8.135兆碱基区域定位了两个已报道的抗病数量性状位点(QTL),即针对番茄斑萎病毒(TSWV)的和针对叶斑病(LS)的。从这个基因组区域开发了719个新的SSR标记,这为这些QTL的精细定位提供了可能。此外,该区域还包含652个基因,其中49个是与防御相关的基因,包括两个NB-ARC基因、三个富含亮氨酸重复序列(LRR)的类受体基因和三个WRKY转录因子。这些抗病相关基因可能有助于花生抵抗病毒(如TSWV)和真菌(如LS)病害。总之,本研究不仅为花生遗传图谱构建和QTL定位提供了大量潜在可用的分子标记,也为基于图谱的基因克隆和分子育种提供了标记。