另外,由于SCoT分子标记扩增片段大小在200~2 000 bp之间[13],而其长度多态性差异通常在几个碱基至几十个碱基之间,利用现有SCoT检测方法(1.5%琼脂糖凝胶电泳或3.5%变性聚丙烯酰胺凝胶电泳)无法精确区分该标记,因此通过对SCoT长度多态性标记片段进行测序分析,可以清楚地了解该片段的序列,并开发出更方便、精确的分子标记。本研究通过对SCoT12分子标记中的一个长度多态性片段进行测序分析,开发出了一个新的SCoT12-SSR分子标记,该SSR标记可以准确区分此位点上的5个不同的等位变异。因此,利用该方法开发新的多态性标记是对现有花生多态性分子标记的补充。 参 考 文 献: [1] Kochert G, Stalker H T, Gimenes M, et al. RFLP and cytogenetic evidence on the origin and evolution of allotetraploid domesticated peanut, Arachis hypogaea (Leguminosae)[J]. Am. J. Bot., 1996, 83: 1282-1291. [2] Kochert G, Halward T, Branch W D, et al. RFLP variability in peanut (Arachis hypogaea L.) cultivars and wild species[J]. Theor. Appl. Genet., 1991, 81: 565-570. [3] McCouch S R, Teytelman L, Xu Y, et al. Development and mapping of 2240 new SSR marker for rice (Oryza sativa L.)[J]. DNA Res., 2002, 9(6): 199-207. [4] Sharopova N, McMullen M D, Linda S, et al. Development and mapping of SSR markers for maize[J]. Plant Mol. Bio., 2002, 48(5/6): 463-481. [5] Rder M S, Korzun V, Wendehake K, et al. A microsatellite map of wheat[J]. Genetics, 1998, 149(4): 2007-2023. [6] Zhao Y, Prakash C S, He G. Characterization and compilation of polymorphic simple sequence repeat (SSR) markers of peanut from public database[J]. BMC Res. Notes, 2012, 5: 362. [7] Guo B, Pandey M K, He G, et al. Recent advances in molecular genetic linkage maps of cultivated peanut[J]. Peanut Science, 2013, 40(2): 95-106. [8] Collard B C Y, Mackill D J. Start codon targeted (SCoT) polymorphism: a simple, novel DNA marker technique for generating gene-targeted markers in plants[J]. Plant Mol. Biol. Rep., 2009, 27: 86-93. [9] Gorji A M, Poczai P, Polgar Z, et al. Efficiency of arbitrarily amplified dominant markers (SCoT, ISSR and RAPD) for diagnostic fingerprinting in tetraploid potato[J]. Am. J. Pot. Res., 2011, 88: 226-237. [10] Xiong F, Zhou R, Han Z, et al. Start condon targeted polymorphism for evaluation of functional genetic variation and relationships in cultivated peanut (Arachis hypogaea L.) genotypes[J]. Mol. Biol. Rep., 2011, 38: 3487-3494. [11] 贺梁琼, 熊发前, 钟瑞春, 等. 利用SCoT标记分析花生栽培种×A. chacoensis组合异源多倍化的早期基因组变化[J]. 中国农业科学, 2013, 46(8): 1555-1563. [12] 熊发前, 唐荣华, 陈忠良, 等. 目标起始密码子多态性(SCoT): 一种基于翻译起始位点的目的基因标记新技术[J]. 分子植物育种, 2009, 7(3): 635-638. [13] 熊发前, 蒋菁, 钟瑞春, 等. 目标起始密码子多态性(SCoT)分子标记技术在花生属中的应用[J]. 作物学报, 2010, 36(10): 2055-2061. 山 东 农 业 科 学 2014,46(11):15~17 |