大豆子叶节遗传转化体系的优化研究  

Optimization for the Transformation System of Soybean Cotyledon Node

作　　者：李冬梅[1] 陈薇[1] 李永光[1] 李文滨[1] LI Dong-mei;CHEN Wei;LI Yong-guang;LI Wen-bin(Soybean Reasearch Institute Key Laboratory of Soybean Biology in Chinese Ministry of Education/Agricultural Ministry＇ s Northeast Areal Key Laboratory of Soybean Biology and Genetic Breeding,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]东北农业大学大豆研究所/大豆生物学教育部重点实验室/农业部东北大豆生物学与遗传育种重点实验室,黑龙江哈尔滨150030

出　　处：《大豆科学》2018年第4期531-538,共8页Soybean Science

基　　金：东北农业大学青年才俊项目(17QC01);抗逆转基因大豆新品种培育专项(2016ZX08004-002)

摘　　要：为了提高大豆转化效率,建立一个稳定高效的大豆遗传转化体系,选择了4个代表性基因型大豆子叶节为外植体,以子叶节和丛生芽GUS瞬时表达率为指标,优化了大豆遗传转化过程。结果表明:在侵染阶段分别进行超声波和抽真空辅助处理,GUS染色显示,4个基因型品种处理不同时间后遗传转化效率均有不同程度提高,超声波30 s或者抽真空2 min后农杆菌的侵染效率提高最为明显,转化效率分别提高8%~42%、16%~41%。在芽诱导前期进行PPT浓度筛选,4种基因型的最适PPT浓度均为0.2 mg·L^(-1),转化效率提高18%~33%。For improving the efficiency and establishing a stable system, the process of soybean cotyledonary transformation was optimized in this study. Cluster buds of cotyledons with four different genotypes were chosen for analyzing the transformation efficiency by b-glueuronidase （GUS） staining. The results indicated that treatments with ultrasonic and vacuum were positive regulators for Agrobacterium infection. GUS staining showed that, the genetic transformation efficiencies of four genotypes were improved after treatment for different periods of time. The preferred conditions for both treatments were ultrasonic for 30 s and vacuum for 2 min, and conversion efficiency increased by 8%-42% and 16%-41%, respectively, which applied for all of the above four genotypes. At the earlier stage of induction of clustered buds, the optimal screening concentration of Phosphinothricin （PPT） was 0. 2 mg·L-1 , and the conversion efficiency increased by 18%-33%.

关 键 词：农杆菌介导 大豆子叶节 超声波 抽真空 

分 类 号：S565.1[农业科学—作物学]