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Identification and Evaluation of Single-Nucleotide Polymorphisms in Allotetraploid Peanut (Arachis hypogaea L.) Based on Amplicon Sequencing Combined with High Resolution Melting (HRM) Analysis

文献类型： 外文期刊

作者： Hong, Yanbin 1 ; Pandey, Manish K. 3 ; Liu, Ying 1 ; Chen, Xiaoping 1 ; Liu, Hong 4 ; Varshney, Rajeev K. 3 ; Liang, X 1 ;

作者机构： 1.Guangdong Acad Agr Sci, Crops Res Inst, Peanut Res Ctr, Guangzhou, Guangdong, Peoples R China

2.Sun Yat Sen Univ, Sch Life Sci, Guangzhou 510275, Guangdong, Peoples R China

3.Int Crops Res Inst Semi Arid Trop, Ctr Excellence Genom, Hyderabad, Andhra Pradesh, India

4.South China Agr Univ, Coll Agr, Guangzhou, Guangdong, Peoples R China

5.Univ Western Australia, Sch Plant Biol, Crawley, WA, Australia

6.Univ Western Australia, Inst Agr, Crawley, WA, Australia

关键词： SNPs;peanut;tetraploid;high resolution melting (HRM);polymorphism

期刊名称：FRONTIERS IN PLANT SCIENCE （ 影响因子：5.753； 五年影响因子：6.612 ）

ISSN： 1664-462X

年卷期： 2015 年 6 卷

页码：

收录情况： SCI

摘要： The cultivated peanut (Arachis hypogaea L.) is an allotetraploid (AABB) species derived from the A-genome (Arachis duranensis) and B-genome (Arachis ipaensis) progenitors. Presence of two versions of a DNA sequence based on the two progenitor genomes poses a serious technical and analytical problem during single nucleotide polymorphism (SNP) marker identification and analysis. In this context, we have analyzed 200 amplicons derived from expressed sequence tags (ESTs) and genome survey sequences (GSS) to identify SNPs in a panel of genotypes consisting of 12 cultivated peanut varieties and two diploid progenitors representing the ancestral genomes. A total of 18 EST-SNPs and 44 genomic-SN Ps were identified in 12 peanut varieties by aligning the sequence of A. hypogaea with diploid progenitors. The average frequency of sequence polymorphism was higher for genomic-SNPs than the EST-SNPs with one genomic-SNP every 1011 bp as compared to one EST-SNP every 2557 bp. In order to estimate the potential and further applicability of these identified SNPs, 96 peanut varieties were genotyped using high resolution melting (HRM) method. Polymorphism information content (PIG) values for EST-SNPs ranged between 0.021 and 0.413 with a mean of 0.172 in the set of peanut varieties, while genomic-SNPs ranged between 0.080 and 0.478 with a mean of 0.249. Total 33 SNPs were used for polymorphism detection among the parents and 10 selected lines from mapping population Y13Zh (Zhenzhuhei x Yueyou13). Of the total 33 SNPs, nine SNPs showed polymorphism in the mapping population Y13Zh, and seven SNPs were successfully mapped into five linkage groups. Our results showed that SNPs can be identified in allotetraploid peanut with high accuracy through amplicon sequencing and HRM assay. The identified SN Ps were very informative and can be used for different genetic and breeding applications in peanut.

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