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Selection and establishment of high oleic acid content in soybean by using their molecular markers

利用分子标记筛选和建立高油酸大豆品种

基本信息

批准号：
11660006
负责人：
TAKAGI Yutaka
金额：
$ 2.5万
依托单位：
Saga University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11660006/
关键词：
soybean fatty acid fatty acid desaturase oleic acid FAD2-1 FAD2-2 ダイス

项目摘要

The mutants KK21 and HOLL, both of which contain 40 to 45% oleic acid of total fatty acids, were reciprocally crossed in our laboratory. Six high oleic acid lines (approx. 60% of total fatty acid) were selected from F2 progeny and were further confirmed by analyzing the fatty acid contents in both of F3 and F4 progenies.As a result, three of the six selected lines showed higher oleic acid phenotype, compared with their parental lines. The oleic acid content of these three lines distributed in the range of 57.9 to 67.6%, 56.4 to 68.4% and 47.4 to 68.4%, respectively. These values were relatively higher than each of their parents, KK21 (37.2 to 51.2%) or HOLL (44.6 to 53.8%), which were grown under same condition. Furthermore, we selected 3 to 5 F3 offspring-lines from each of 3 lines and those 11 offspring-lines that consist of 20 plants of F4 progeny, were subjected to analyze for oleic acid content. We found one of these lines that has very high oleic acid content which is more than 60%.In addition, we carried out a comparison study for the expression levels of FAD2-1 or FAD2-2 with northern-blot and quantitative RT-PCR techniques, but no significant difference was detected between Bay (original variety) and other mutants. To determine their mutation at the base substitution level, we are now analyzing the nucleotide sequences of FAD2-1 and FAD2-2 gene, that were isolated from Bay and other mutants.It is necessary to push forward the examination of genetic stability of very high oleic acid content of selected one using the next generation, and to analyze the genotype of this selected germline with the designed primers that can detect the polymorphism between Bay and other mutants.

我们对油酸含量占总脂肪酸40 ~ 45%的突变体KK 21和HOLL进行了回交。6条高油酸生产线（约通过对F3和F4代脂肪酸含量的分析，进一步证实了F2代脂肪酸含量高（占总脂肪酸的60%），其中3个品系的油酸表型高于其亲本。3个品系的油酸含量分别为57.9 ~ 67.6%、56.4 ~ 68.4%和47.4 ~ 68.4%。这些值相对高于在相同条件下生长的亲本KK 21（37.2 - 51.2%）或HOLL（44.6 - 53.8%）。此外，我们从3个品系中的每一个中选择3至5个F3后代品系，并且对由20株F4后代植物组成的这11个后代品系进行油酸含量分析。利用Northern blot和定量RT-PCR技术对FAD 2 -1和FAD 2 -2的表达水平进行了比较研究，结果表明，Bay与其他突变体的表达水平差异不显著。为了在碱基替换水平上确定它们的突变，我们现在正在分析从Bay和其他突变体中分离的FAD 2 -1和FAD 2 -2基因的核苷酸序列。有必要利用下一代推进对所选择的极高油酸含量的遗传稳定性的检查，并利用设计的能检测Bay与其它突变体之间多态性的引物对该品系进行基因型分析。