小麦HMW-GS检测方法的优化及应用  

Optimization and Application of the Detection Method of HMW-GS in Common Wheat

作　　者：高正[1] 王晓龙[1] 张晓科[1] 白升升[1] 付晓洁[1] 相吉山[2,3] 穆培源[2,3] 梁强[1] 

机构地区：[1]西北农林科技大学农学院/国家小麦改良中心杨凌分中心,陕西杨凌712100 [2]新疆农垦科学院作物研究所,新疆石河子832000 [3]谷物品质与遗传改良兵团重点实验室,新疆石河子832000

出　　处：《麦类作物学报》2013年第5期929-934,共6页Journal of Triticeae Crops

基　　金：新疆兵团十二五主要生物种质资源研究专项(2012BB047);国家小麦产业技术体系建设专项(CARS-03);西北农林科技大学唐仲英育种基金项目

摘　　要：为了准确快速鉴定小麦品种的HMW-GS组成,以18个已知HMW-GS组成的品种为对照,将4种不同浓度分离胶的SDS-PAGE与分子标记(Bx7、Bx7OE和By 8)相结合,构建一套适于检测HMW-GS组成的方法,并用该方法检测214份陕西小麦品种(系)的HMW-GS组成。4种分离胶浓度中,除了亚基7、7*与7OE和8与8*外,9%的分离胶可以很清楚地分离Glu1-位点的其他常见亚基,结合分子标记(Bx7、Bx7OE和By 8)检测对照品种,结果与已知亚基(基因)一致。用分离胶浓度为9%的SDS-PAGE结合分子标记(Bx7、Bx7OE和By 8)方法对陕西品种(系)检测结果表明,Glu-A1、Glu-B1和Glu-D1分别有3、8和3个亚基种类,共28种亚基组合类型。以1、Null、7+9、7+8、7+8*、14+15、2+12和5+10为主要亚基类型,频率分别为55.6%、41.6%、43.9%、26.2%、10.3%、15.4%、79.0%和12.6%;同时在Glu-B1位点发现7+8*和6+8*亚基。该方法可以快速准确地评价小麦HMW-GS组成,有效区分Glu-B1位点的亚基7与7OE,8与8*,鉴定结果稳定可靠。High molecular weight glutenin subunit （HMW GS） composition is closely related to processing quality of wheat. It is very important to identify HMW GS composition for wheat quality improvement. A method, based on using four different concentrations of separation gel of SDS PAGE and molecular markers on Bx7, Bx7OE and By 8, was developed and validated by 18 cotrol cultivars with known HMW GS.Among four concentrations of separation gel, 9% separation gel can clearly separated most HMW GS except the subunits 7, 7OE ,8＊ and 8,which were distinguished with molecular markers. The identifying results of cultivars were consistent with their known HMW GS. It indicated that the method was stable and reliable. Then the HMW GS compositions of 214 major cultivars from Shaanxi Province were identified by the developed method. There were 28 HMW GS combinations detected in the 214 wheat cultivars. Three, eight and three alleles were present at Glu A1, Glu B1and Glu D1 loci, respectively in wheat from Shaanxi Province. The most common alleles were 1, Null, 7＋9, 7＋8, 7＋8＊, 14＋15, 2＋12 and 5＋10, and their frequencies were 55.6%, 41.6%, 43.9%, 26.2%, 10.3%, 15.4%, 79.0% and 12.6%, respectively. HMW GS 7＋8＊ and 6＋8＊ were found in Shaanxi wheat. In conclusion, most HMW GS, especially 7 and 7OE, 8 and 8＊ at Glu B1 locus, could be effectively distinguished by the developed method. In this study, more HMW GS were detected in Shaanxi wheat cultivars, but few HMW GS with strong gluten were found. Combining the SDS PAGE and molecular markers may serve as a rapid and efficient method to cultivar quality evaluation in wheat breeding.

关 键 词：小麦 HMW GS SDS PAGE 分子标记 

分 类 号：S512.1[农业科学—作物学] S330