JI Cai-feng,SHAO Rui-chao,ZHANG Qi,et al.Molecular Detection and Resistance Analysis of Soybean-lines with HrpZPsta[J].Soybean Science,2020,39(02):220-226.[doi:10.11861/j.issn.1000-9841.2020.02.0220]
转hrpZPsta大豆株系的分子检测和抗性分析
- Title:
- Molecular Detection and Resistance Analysis of Soybean-lines with HrpZPsta
- 关键词:
- 大豆; JN27-119-21; 疫霉根腐病; hrpZPsta基因; 抗病性
- Keywords:
- Soybean; JN27-119-21; Phytophthora root rot; hrpZPsta Gene; Disease resistance
- 文献标志码:
- A
- 摘要:
- 为考察转基因大豆株系JN27-119-21后代中hrpZPsta外源基因的遗传稳定性、检测JN27-119-21后代的抗病能力，从而为培育抗疫霉根腐病的大豆新品种提供理论参考，以转hrpZPsta基因大豆株系JN27-119-21的T7、T8代为供试材料，通过PCR和Southern Blot分子生物学检测方法和下胚轴侵染法鉴定转基因株系后代抗疫霉菌能力。研究结果表明：外源hrpZPsta基因在高世代转基因株系中能够稳定遗传，hrpZPsta基因已被成功整合到大豆受体吉农27的基因组中，且整合方式为单拷贝，整合位点不同。hrpZPsta基因在转化株系的根、茎、叶中均有表达，T7代平均相对表达量分别为2.877，1.336和7.734，T8代平均相对表达量分别为3.612、1.746和8.627，2个世代相对表达量水平均为叶＞根＞茎。转基因株系后代对疫霉根腐病的抗病级别为抗性，而受体株系为中抗，抗病能力明显提高。
- Abstract:
- In order to verify the genetic stability of hrpZPsta gene in transgenic line JN27-119-21 and detect the disease resistance of the progenies of JN27-119-21, so as to provide a theoretical reference for breeding new soybean varieties resistant to Phytophthora root rot, in this experiment, T7 and T8 generation of soybean-lines JN27-119-21 with hrpZPsta gene were used as test materials for identificating the disease resistance ability by PCR and Southern Blot molecular biological detection and hypocotyl infection to identify the resistance to Phytophthora of the offspring of JN27-119-21. The results showed: The exogenous hrpZPsta gene can be stably inherited in high-generation transgenic lines, the hrpZPsta gene was successfully integrated into the genome of soybean receptor Jinong 27 with single copy, and the integration sites were different. The hrpZPsta gene was expressed in the roots, stems and leaves of the transformed lines, the average relative expression of the T7 was 2.877, 1.336 and 7.734, the average relative expression of T8 was 3.612, 1.746 and 8.627, and the relative expression level the T7 and T8 were both leaf>root>stem. The offspring of the transgenic lines were resistant to the disease resistance level of Phytophthora root rot, while the recipient strain was medium resistant and the disease resistance was significantly improved.
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备注/Memo
收稿日期：2019-09-27