Isolation and Molecular Marker Detection of Badh2 Gene from Aromatic Rice Germplasm Resources in Southern Henan

Bo Peng, Yu Zhu, Zi-yu Wang, Juan Peng, Lu-Lu He, Xiayu Tian, Ziyue Liu, Ling Sun, Ying Wang, Xiao-Hua Song, Huilong Li, Yan-Yang Sun, Rui-hua Pang, Jintao Li, Quanxiu Wang, Wei Zhou, Hongyu Yuan
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Abstract

The production of aroma in aromatic rice is due to the increase of 2-acetyl-1-pyrroline (2-AP) precursor substances caused by the functional deletion of Badh2 gene on chromosome 8, and the accumulation of 2AP makes rice produce aroma. In this study, Badh2 gene was isolated and cloned from 18 representative aromatic rice cultivars in Southern Henan, and the bioinformatics analysis of Badh2 gene was carried out. Meanwhile, seven functional molecular markers developed by Badh2 gene were used to detect and analyze Badh2 gene in 18 aromatic rice varieties from Southern Henan. The results showed that the coding region of Badh2 gene was 1509 bp in length. It contained 15 exons and 14 introns, and encoded 503 amino acids. There are many types of variation of the Badh2 gene in the 18 aromatic rice varieties. According to the variation of Badh2 gene, the tested aromatic rice varieties could be divided into three groups, among which Xinxianggeng 1, Xiangnuo 25, Heixiangdao 193 and Xiangbao 2 were concentrated in group Ⅰ, while the other 14 kinds of aromatic rice were concentrated in group II. Seven functional molecular markers of Badh2 gene were used to detect different varieties mutation types in exon 2, exon 4~5, exon 7 and exon 13 of Badh2 gene. No aromatic rice varieties with different mutation types were found in promoter region, exon 12 and exon 14 of Badh2 gene. Therefore, our results provide important information for understanding the genetic basis of fragrant genes in aromatic rice germplasm resources in Southern Henan and breeding new varieties of high-quality aromatic rice using molecular marker-assisted selection.
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豫南香稻种质资源中Badh2基因的分离及分子标记检测
芳香水稻香气的产生是由于8号染色体上Badh2基因的功能性缺失导致2-乙酰基-1-吡咯啉(2-AP)前体物质的增加,2AP的积累使水稻产生香气。本研究从豫南18个代表性香稻品种中分离并克隆了Badh2基因,并对Badh2基因进行了生物信息学分析。同时,利用Badh2基因开发的7个功能分子标记,对18个豫南香稻品种的Badh2基因进行了检测和分析。结果表明,Badh2基因的编码区长度为1509 bp。它包含15个外显子和14个内含子,编码503个氨基酸。在18个香稻品种中,Badh2基因存在多种变异类型。根据Badh2基因的变异,所试香稻品种可分为3个群体,其中新香更1号、香诺25号、黑香道193号和香宝2号集中在Ⅰ群体,其余14种香稻集中在II群体。利用Badh2基因的7个功能分子标记检测了Badh2基因外显子2、外显子4~5、外显子7和外显子13的不同变异类型。在Badh2基因启动子区、12外显子和14外显子未发现不同突变类型的芳香水稻品种。因此,本研究结果为了解豫南香稻种质资源中芳香基因的遗传基础,以及利用分子标记辅助选择选育优质香稻新品种提供了重要信息。
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