The fragrance of rice is one of the premium characteristics that breeders want to include in rice varieties due to the higher market value. Nucleotide deletions in exons 2(7 bp) and 7(8 bp) of Betaine Aldehyde Dehydro...The fragrance of rice is one of the premium characteristics that breeders want to include in rice varieties due to the higher market value. Nucleotide deletions in exons 2(7 bp) and 7(8 bp) of Betaine Aldehyde Dehydrogenase 2(BADH2) are associated with fragrance in rice. In this study, a new 13 bp deletion in exon 7 of the BADH2 gene was discovered in the Nang Thom Cho Dao(NTCD) variety, and the mutation has been closely related to the genetic background of indica subspecies through the Bayesian phylogenetic approach and haplotype network analysis of the 3 000 Rice Genomes Project. In addition, a set of functional markers(EX07-13F, EX07-13RN, and EX07-13RM) identified the 13 bp deletion only within NTCD(no amplified band) compared with both non-aromatic and other aromatic rice varieties(110 bp band). The deletion of 13 bases instead of 8 bases in exon 7 of BADH2 caused a premature stop codon, which down-regulated the expression of the BADH2 transcript while associated with up-regulation of OsP5CS and the high amount of 2-acetyl-1-pyrroline. It is potential to use the deletion in exon 7 of the BADH2 gene as a novel marker for adulteration and breeding of fragrant rice varieties, particularly for NTCD.展开更多
In this study,we performed amplificaion and sequence analysis of exon7 of gene Badh2 of 12 fragrant rice materials,and identified the aroma of fragrant rice materials by the method of seed chewing and KOH soaking,so a...In this study,we performed amplificaion and sequence analysis of exon7 of gene Badh2 of 12 fragrant rice materials,and identified the aroma of fragrant rice materials by the method of seed chewing and KOH soaking,so as to analyze the sequence mutation of exon 7 in the Badh2 gene of rice material and its corresponding relation with the flavor character.The results showed that an 8 bp deletion(aaaa--t---ggc)and a mutation of SNP(g→t)in exon 7 of Badh2 gene were found in 10 materials,including Xiangnuo,Lvjinxiang,Meixiangzhan 2,Huaxiang,Yuexiangxuan 1,Hongyuxiang,Meixiangxuan 1,Baxiangxuan 1,Taixiangxuan 1,Taixiangxuan 2.This mutation was consistent with the mutation of EU155083 sequence in GenBank and was reported for the first time in Chinese rice materials.In these 10 fragrant rice materials with mutation,Huaxiang and Meixiangxuan 1 were identified as the heterozygote genotype,and Hongyuxiang was identified as non-fragrant rice,so the sequence mutation in exon 7 of Badh2 gene in fragrant rice materials did not correspond to aroma traits one by one;and 7 materials were identified as fragrant rice,and the brown rice of Meixiangzhan 2 and Yuexiangyuan 1 had sweet taste.The results could provide a reference for the research on the genetic mechanism of rice aroma character and the promotion of fragrant rice varieties.展开更多
badh2是控制水稻香味的主效基因,其与等位非香基因Badh2功能差异主要在于第七外显子上的8 bp缺失及3个SNP,开发其易于分辨且稳定的功能标记将有助于香稻分子育种。本研究基于四引物扩增受阻突变体系PCR(Tetra Primer Amplification Refr...badh2是控制水稻香味的主效基因,其与等位非香基因Badh2功能差异主要在于第七外显子上的8 bp缺失及3个SNP,开发其易于分辨且稳定的功能标记将有助于香稻分子育种。本研究基于四引物扩增受阻突变体系PCR(Tetra Primer Amplification Refractory Mutation System PCR,Tetra-primer ARMS PCR)原理,在功能差异位点外侧两引物一致的情况下,分别设计与badh2和Badh2完全匹配的5'端添加错配碱基的靶引物。结果表明,经1%的琼脂糖凝胶电泳后,由完全匹配引物组成的PCR体系基本能分辨badh2/Badh2基因型,但目标条带较模糊;而引物中5'端引入错配碱基的PCR体系能产生更明晰、易于分辨的目标条带。本研究将有助于香稻分子育种及为其他类似分子标记的优化提供借鉴。展开更多
基金funded in part by the Can Tho University Improvement Project VN14-P6 supported by a Japanese Official Development Assistance loan。
文摘The fragrance of rice is one of the premium characteristics that breeders want to include in rice varieties due to the higher market value. Nucleotide deletions in exons 2(7 bp) and 7(8 bp) of Betaine Aldehyde Dehydrogenase 2(BADH2) are associated with fragrance in rice. In this study, a new 13 bp deletion in exon 7 of the BADH2 gene was discovered in the Nang Thom Cho Dao(NTCD) variety, and the mutation has been closely related to the genetic background of indica subspecies through the Bayesian phylogenetic approach and haplotype network analysis of the 3 000 Rice Genomes Project. In addition, a set of functional markers(EX07-13F, EX07-13RN, and EX07-13RM) identified the 13 bp deletion only within NTCD(no amplified band) compared with both non-aromatic and other aromatic rice varieties(110 bp band). The deletion of 13 bases instead of 8 bases in exon 7 of BADH2 caused a premature stop codon, which down-regulated the expression of the BADH2 transcript while associated with up-regulation of OsP5CS and the high amount of 2-acetyl-1-pyrroline. It is potential to use the deletion in exon 7 of the BADH2 gene as a novel marker for adulteration and breeding of fragrant rice varieties, particularly for NTCD.
基金Earmarked Fund for China Agriculture Research System(CARS-01-89)Fundamental Scientific Research Fund of Hainan Academy of Agricultural Sciences(JBKYYWF2020-03)Science and Technology Innovation Project of Hainan Academy of Agricultural Sciences(KJCX-2020-11)。
文摘In this study,we performed amplificaion and sequence analysis of exon7 of gene Badh2 of 12 fragrant rice materials,and identified the aroma of fragrant rice materials by the method of seed chewing and KOH soaking,so as to analyze the sequence mutation of exon 7 in the Badh2 gene of rice material and its corresponding relation with the flavor character.The results showed that an 8 bp deletion(aaaa--t---ggc)and a mutation of SNP(g→t)in exon 7 of Badh2 gene were found in 10 materials,including Xiangnuo,Lvjinxiang,Meixiangzhan 2,Huaxiang,Yuexiangxuan 1,Hongyuxiang,Meixiangxuan 1,Baxiangxuan 1,Taixiangxuan 1,Taixiangxuan 2.This mutation was consistent with the mutation of EU155083 sequence in GenBank and was reported for the first time in Chinese rice materials.In these 10 fragrant rice materials with mutation,Huaxiang and Meixiangxuan 1 were identified as the heterozygote genotype,and Hongyuxiang was identified as non-fragrant rice,so the sequence mutation in exon 7 of Badh2 gene in fragrant rice materials did not correspond to aroma traits one by one;and 7 materials were identified as fragrant rice,and the brown rice of Meixiangzhan 2 and Yuexiangyuan 1 had sweet taste.The results could provide a reference for the research on the genetic mechanism of rice aroma character and the promotion of fragrant rice varieties.
文摘badh2是控制水稻香味的主效基因,其与等位非香基因Badh2功能差异主要在于第七外显子上的8 bp缺失及3个SNP,开发其易于分辨且稳定的功能标记将有助于香稻分子育种。本研究基于四引物扩增受阻突变体系PCR(Tetra Primer Amplification Refractory Mutation System PCR,Tetra-primer ARMS PCR)原理,在功能差异位点外侧两引物一致的情况下,分别设计与badh2和Badh2完全匹配的5'端添加错配碱基的靶引物。结果表明,经1%的琼脂糖凝胶电泳后,由完全匹配引物组成的PCR体系基本能分辨badh2/Badh2基因型,但目标条带较模糊;而引物中5'端引入错配碱基的PCR体系能产生更明晰、易于分辨的目标条带。本研究将有助于香稻分子育种及为其他类似分子标记的优化提供借鉴。
