香稻品种OsBADH2突变位点分析及其功能标记的开发

doi:10.13560/j.cnki.biotech.bull.1985.2020-1086

摘要/Abstract

摘要：

香稻拥有浓郁的香味和较高的营养价值,深得消费者的喜爱和育种专家的重视,挖掘和利用香稻基因资源为培育优质香稻品种奠定基础。水稻甜菜碱醛脱氢酶基因（OsBADH2）功能的丧失导致稻米具有香味。通过对10份香稻品种OsBADH2编码区进行测序分析,明确其变异类型。根据香稻OsBADH2第7外显子8 bp的缺失,设计功能标记E7,利用E7分别对测序的10份香稻品种和2个香稻/非香稻育种群体的OsBADH2基因型进行验证与检测。结果表明,象牙香占、花香B、农香24等9份香稻均在第7外显子发生8 bp的缺失和3 bp的替换,而香稻Xiao7在第14外显子有1个碱基（G）的插入突变。功能标记E7能快速准确地鉴定OsBADH2的纯合显性、纯合隐性和杂合基因型。

关键词: 香稻, OsBADH2, 功能标记, 分子标记辅助选择

Abstract:

Fragrant rice is favored worldwide by consumers and rice breeders,due to its full aroma and high nutritional value. Exploring and utilizing the flagrant gene resources would lay a foundation for breeding excellent-quality varieties. Loss of function of the betaine aldehyde dehydrogenase gene（OsBADH2）leads to that rice is of fragrance. OsBADH2 coding regions in 10 fragrant rice varieties were sequenced to explicit its mutant type. A functional marker E7 was designed according to the 8-bp deletion in exon 7 of OsBADH2 in the fragrant rice. Genotypes of 10 fragrant rice varieties and 2 populations derived from fragrant rice/non-fragrant varieties were identified and verified by E7. The results indicated that 8 bp deletion and 3 bp replacement in the exon 7 of OsBADH2 gene were found in 9 cultivars,including Xiangyaxiangzhan,Huaxiang B and Nongxiang 24 et a1. And 1 bp（G）insertion mutation in exon 14 in the fragrant rice of Xiao7 appeared. The homozygous dominant,homozygous recessive and heterozygous genotypes of OsBADH2 could be rapidly and accurately distinguished by functional marker E7.

Key words: fragrant rice, OsBADH2, functional marker, molecular marker-assisted selection

孙平勇, 张武汉, 舒服, 何强, 张莉, 彭志荣, 邓华凤. 香稻品种OsBADH2突变位点分析及其功能标记的开发[J]. 生物技术通报, 2021, 37(4): 1-7.

SUN Ping-yong, ZHANG Wu-han, SHU Fu, HE Qiang, ZHANG Li, PENG Zhi-rong, DENG Hua-feng. Analysis of Mutation Sites of OsBADH2 Gene in Fragrant Rice and Development of Related Functional Marker[J]. Biotechnology Bulletin, 2021, 37(4): 1-7.

图/表 6

图1 OsBADH2 编码区的扩增

Fig. 1 Amplification result from coding regions of OsBADH2

图2 OsBADH2 的基因结构和香稻的变异类型

Fig. 2 Genetic structure of OsBADH2 and allelic variation types in fragrant rice

图3 功能标记E7 在OsBADH2 上的位置

Fig.3 Location of functional marker E7 in the OsBADH2

图 4 功能标记E7 对不同品种OsBADH2 基因型的鉴定 M：100 bp marker;1-12：巴斯马蒂、香杂1 号、玉油香占、象牙香占、花香B、农香24、稻花香、香粳、明香10S、xiao7、魔王谷和02428

Fig. 4 Molecular detection of OsBADH2 genotypes by functional marker E7 in different rice varieties M: 100 bp marker. 1-12: Basmati, Xiangza 1, Yuyouxiangzhan, Xiangyaxiangzhan, Huaxiang B, Nongxiang 24, Daohuaxiang, Xianggeng, Mingxing10S, xiao7, Mowanggu, and 02428

图5 不同品种香味物质2AP含量比较 a、b、c 表示显著性差异

Fig. 5 Content of 2AP in different rice varieties a, b and c represent significant differences

图6 功能标记E7 对2 份育种群体OsBADH2 基因型的鉴定 M: 100 bp marker;(A) 1: 稻花香;2: 粳944;3-30: 粳944/ 稻花香// 粳944 群体的部分单株。(B) 1: 明香10S;2: 泰丰B;3-30: 泰丰B/ 明香10S// 泰丰B 群体的部分单株

Fig. 6 Molecular detection of OsBADH2 genotypes by functional marker E7 in two breeding populations M: 100 bp marker. (A) 1: Daohuaxiang. 2: Geng 944, 3-30: Parts of plants from Geng 944 / Daohuaxiang//Geng 944 population. (B) 1: Mingxing10S, 2: Taifeng B. 3-30: Parts of plants from Taifeng B/Mingxing10S // Taifeng B

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