小麦TaP5CS基因抗旱相关分子标记的开发  

Development of Molecular Marker for TaP5CS Gene for Improving Drought Resistance in Wheat

作　　者：于铭 魏凡 常鹏举 杨智全[1] 武军[1] 陈东升[2] 张晓科[1] YU Ming;WEI Fan;CHANG Pengju;YANG Zhiquan;WU Jun;CHEN Dongsheng;ZHANG Xiaoke(College of Agronomy,Northwest A&F University,Yangling,Shaanxi 71200,China;The Crop Research Institute,Ningxia Academy of Agriculture and Forestry Science,Yinchuan,Ningxia 750002,China)

机构地区：[1]西北农林科技大学农学院,陕西杨凌712100 [2]宁夏农林科学院农作物研究所,宁夏银川750002

出　　处：《麦类作物学报》2021年第6期658-664,共7页Journal of Triticeae Crops

基　　金：国家重点研发计划项目(2016YFD0101802);国家自然科学基金项目(31671693);陕西省重点研发计划项目(2019ZDLNY04-05);宁夏农业育种专项(NXNYYZ201302);中央引导地方科技发展专项(2020YDDF0056);宁夏农林科学院科技先导资金项目(DWX-2019001)。

摘　　要：脯氨酸是小麦抵御干旱胁迫的重要调节物质,吡咯啉-5-羧酸合成酶(Pyrrpline-5-carboxylatesynthetase,P5CS)是干旱条件下脯氨酸合成的关键酶。为了提高小麦抗旱品种的选育效率,本研究选用国内119个小麦品种(系),鉴定其萌发期抗旱性;并从中选取10个抗旱性不同的小麦品种,分析响应干旱的叶片脯氨酸积累量与其抗旱性的关系;通过两组极端抗旱品种间TaP5CS基因序列比对和随机群体验证,开发该基因抗旱相关分子标记。结果表明,119个小麦品种(系)的抗旱等级不同,部分品种间相对发芽率的差异达到极显著水平,响应干旱的叶片脯氨酸积累量与抗旱性呈正相关。TaP5CS-1A基因在两组极端品种间基因组序列存在2个单核苷酸变异(SNP)位点,扩增第5内含子上涵盖SNP位点的1894 bp序列,其中2个强抗旱性品种在此SNP位点不能被PflmⅠ酶切,命名为TaP5CS-1A-a等位变异;2个弱抗旱性品种在此SNP位点被酶切为1706 bp和188 bp的条带,命名为TaP5CS-1A-b等位变异,以此开发分子标记TaP5CS-1A-CAPS。利用此标记检测119个小麦品种(系),采用普通PCR和半巢式PCR的方法分别进行扩增和相同酶切,结果完全一致:携带TaP5CS-1A-a和TaP5CS-1A-b等位变异类型的品种分别有61和58个,平均相对发芽率分别为60.7%和44.6%,前者极显著高于后者(P<0.01),说明TaP5CS-1A基因与小麦抗旱性相关,开发该基因分子标记TaP5CS-1A-CAPS可以用于小麦抗旱性的筛选与鉴定。Proline is an important regulator of wheat resistance to drought stress.Pyrrpline-5-carboxylate synthetase(P5 CS)is a key enzyme for proline synthesis under drought condition.In order to improve the breeding efficiency of drought-resistant wheat varieties,in this study 119 domestic wheat varieties(lines)were selected to identify their drought resistance at germination period.10 varieties with different drought resistance were selected to analyze the relationship between the accumulation of proline in leaf and drought resistance in response to drought.The molecular markers related to drought resistance were developed through sequence alignment and random population validation between two groups of stronger and weaker drought-resistant varieties.The results showed that the relative germination rate of varieties(lines)with different drought resistance levels was significantly different(P<0.01).The proline accumulation in the leaves in response to drought was positively correlated with drought resistance.The TaP5 CS-1 A gene has 2 single nucleotide polymorphism(SNP)sites in the genome sequence between stronger and weaker drought-resistant cultivars,and the 1894 bp sequence covering the SNP site in the fifth intron was amplified.The two stronger drought-resistant cultivars were designated as TaP5 CS-1 A-a allele because they can not be digested by PflmⅠenzyme at this SNP site.The two weaker drought-resistant varieties were enzymatically digested into 1706 bp and 188 bp bands at this SNP site and designated as TaP5 CS-1 A-b allelic variation,so as to develop molecular markers TaP5 CS-1 A-CAPS,which were used to distinguish the allelic variation among the 119 varieties(lines).In order to facilitate the observation of the electrophoresis results,the method of half-nested PCR was used for PCR amplification and the same enzyme digestion,which were completely consistent with the results of ordinary PCR.The number of cultivars with TaP5 CS-1 A-a and TaP5 CS-1 A-b were 61 and 58,respectively,and the average relative germina

关 键 词：普通小麦 抗旱性 TaP5CS 分子标记 

分 类 号：S512.1[农业科学—作物学] S330