Microarray-Based Genetic Mapping Using Soybean Near-Isogenic Lines and Generation of SNP Markers in the Rag1 Aphid-Resistance Interval
Microarray-Based Genetic Mapping Using Soybean Near-Isogenic Lines and Generation of SNP Markers in the Rag1 Aphid-Resistance Interval
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DOI：
10.3835/plantgenome2008.04.0207
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发表时间：
2008-11-01
期刊：
影响因子：
4.2
通讯作者：
Hudson, Matthew E.
中科院分区：
文献类型：
--
作者：
Kaczorowski, Karen A.;Kim, Ki-Seung;Hudson, Matthew E.
利用近等基因系(NILs)和Affymetrix大豆基因芯片微阵列技术，筛选与大豆抗蚜基因Rag1紧密连锁的遗传标记。作者声明：[MER.将抗蚜虫品种Dowling和敏感品种Dwight的基因组DNA标记后与阵列杂交，共检测到1500多个可能的单特征多态。为了寻找与Rag1抗蚜基因座紧密连锁的多态性，还对从Dowling回交到Dwight的两个近等基因系的基因组DNA样品进行了杂交。近等基因系与轮回亲本杂交信号的比较表明，在近等基因系与轮回亲本之间，每个近等基因系都有70多个SFP。两个近等基因系共有22个SFP，代表推测与Rag1连锁的分子标记。选择的4个SFP被转化为SNP标记，经常规遗传作图证实与Rag1紧密连锁。我们描述的技术可以用来识别感兴趣的遗传区域的多态，并使用合适的寡核苷酸微阵列产生与重要农艺性状紧密连锁的分子标记。
A strategy using near-isogenic lines (NILs) and Affymetrix Soybean GeneChip microarrays was employed to identify genetic markers closely linked to the soybean aphid [Aphis glycines Matsumura (Hemiptera: Aphididae)] resistance gene Rag1 in soybean [Glycine max (L.) Merr.]. Genomic DNA from the aphid-resistant cultivar Dowling and the aphid-susceptible cultivar Dwight was labeled and hybridized to arrays, identifying more than 1500 putative single feature polymorphisms (SFPs) between these genotypes. To find polymorphisms closely linked to the Rag1 aphid-resistance locus, genomic DNA samples from two NILs developed through backcrossing Rag1 from Dowling four times to Dwight were also hybridized. Comparison of hybridization signals between the NILs and the recurrent parent identified more than 70 SFPs in each NIL between the NIL and the recurrent parent genotype. There were 22 SFPs shared by both NILs, representing molecular markers putatively linked to Rag1. Four selected SFPs were converted to SNP markers and confirmed by conventional genetic mapping to be closely linked to Rag1. The technique that we describe can be used to identify polymorphisms in a genetic region of interest and generate molecular markers closely linked to an agronomically important trait using a suitable oligonucleotide microarray.