小麦籽粒2A染色体PPO基因新分子标记的开发与应用  

A New STS Marker for Grain PPO Gene on 2A chromosome in Common Wheat

作　　者：周志良[1] 司红起[1] 闫小燕[1] 徐恩静[1] 常成[1] 张海萍[1] 卢杰[1] 马传喜[1] 

机构地区：[1]农业部安徽省小麦区域技术创新中心,安徽小麦产业工程技术研究中心,省部共建分子植物育种重点实验室,安徽农业大学农学院,合肥230036

出　　处：《分子植物育种》2011年第1期81-86,共6页Molecular Plant Breeding

基　　金：现代农业产业技术体系专项经费;国家科技支撑计划(2009BADA6B01);安徽省高校自然科学研究重点项目;安徽省良种优质化专项(08010301065);安徽农业大学科研启动基金(wd2008-10)共同资助

摘　　要：参照位于小麦2B染色体上的多酚氧化酶(Polyphenol Oxidase)基因保守区序列(GenBank:AB254804)设计引物,以一组籽粒PPO活性两极端材料为扩增模板进行筛选,并以195份中国微核心种质材料对其有效性验证,旨在开发小麦籽粒PPO基因新分子标记。在设计的引物中,编号为F-4的引物PCR产物在籽粒PPO活性处于两极端的12份小麦品种中表现出规律性极强的多态性,在籽粒PPO活性极高的6个小麦品种扩增出了1条带(a),在籽粒PPO活性极低的6个小麦品种扩增出了3条带(a,b,c)。利用195个微核心种质材料对引物F-4进行有效性验证,其中123个材料扩增出了一条带(a带),其PPO均值为246.22A475U/(min·g·103),剩下的72个材料都扩增出3条带(a,b,c);其PPO活性均值为155.95A475U/(min·g·103);比前者低90.27A475U/(min·g·103),差异显著。在此基础上利用一套中国春缺体将b条带定位于2A染色体上。扩增序列Blast分析发现,b序列与EF070148序列(2Ab)重合区域为99%,相似达99%,基本可以认定为同一序列;a序列与EF070149序列(2Da)重合区域达99%,相似为98%,基本可以认定为同一序列。位于小麦2A染色体上的PPO基因分子标记F-4可以在小麦PPO活性分子育种中加以应用。Based on a wheat grain DNA sequence AB254804（GenBank Accession Number） mapped to chromosome 2B,some pairs of primers were designed.A group of wheat cultivars with high and low PPO activity were amplified to find PPO gene alleles.The six cultivars with the three fragments（a,b,c） had significantly lower mean PPO activity than those with single fragments（Sequence of a）.A total of 195 cultivars were used to verify the association between PPO activities and the STS marker.In the present study,we developed a gene-specific marker（F-4） for low PPO activity.Three fragments were detected in the cultivars with low PPO activity 155.95 A475U/（min·g·103） and a single PCR fragment（about 400 bp） in the cultivars with high PPO activity 246.22 A475U/（min·g·103）.The marker F-4 was mapping to chromosome 2A using a set of nulli-tetrasomic lines of Chinese Spring.Through blast search in NCBI,we found that Sequence of b is very similar to EF070148（2Ab） which was positioned on the 2A chromosome,and sequence of a is very similar to EF070149（2Db） which was positioned on 2D chromosome.Thus,the STS markers F-4 is an efficient and reliable markers for the evaluation of PPO activity and can be used in wheat breeding programs.

关 键 词：小麦 多酚氧化酶 内含子 等位变异 

分 类 号：S512.1[农业科学—作物学]