[1]姬月梅,陈受宜,李英慧,等.农杆菌介导大豆子叶节遗传转化体系的优化研究[J].大豆科学,2008,27(01):26-32.[doi:10.11861/j.issn.1000-9841.2008.01.0026]
　JI Yue-mei,CHEN Shou-yi,LI Ying-hui,et al.Optimization of Genetic Transformation System from Soybean Cotyledon Mediated by Agrobacterium[J].Soybean Science,2008,27(01):26-32.[doi:10.11861/j.issn.1000-9841.2008.01.0026]
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农杆菌介导大豆子叶节遗传转化体系的优化研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第27卷 期数: 2008年01期 页码: 26-32 栏目: 出版日期: 2008-02-25
Title:
Optimization of Genetic Transformation System from Soybean Cotyledon Mediated by Agrobacterium
作者:
姬月梅1; 2; 陈受宜3; 李英慧2; 张丽娟2; 宋晓华1; 常汝镇2; 邱丽娟2
1宁夏大学农学院，宁夏银川 750021;
2中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程/农业部作物种质资源与生物技术重点开放实验室，北京100081；
3中国科学院遗传与发育生物学研究所，国家重点实验室植物基因组学，北京100101
Author(s):
JI Yue-mei1; 2; CHEN Shou-yi3; LI Ying-hui2; ZHANG Li-juan2; SONG Xiao-hua1; CHANG Ru-zhen2; and QIU Li-juan2
1Ningxia University Agricultural College，Yinchuan 750021，Ningxia;
2The National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI) / Key Laboratory of Germplasm & Biotechnology (MOA)，Institute of Crop Sciences，Chinese Academy of Agricultural Sciences，100081 Beijing;
3The National Key Lab of Plant Genomics，Institute of Genetics and Developmental Biology，Chinese Academy of Sciences，Beijing 100101，China
关键词:
大豆; 遗传转化; 子叶节; Na+/H+逆向转运蛋白; 再生
Keywords:
Soybean; Transformation; Cotyledonary-node; Na+/H+ antiporter; Regeneration
DOI:
10.11861/j.issn.1000-9841.2008.01.0026
文献标志码:
A
摘要:
以大豆品种“中豆32、Peking、早熟18、绥农14”子叶节为受体材料，用农杆菌介导法转入与抗逆相关的小麦Na+/H+逆向转运蛋白基因（TaNHX2），探索外植体大小、培养基主要成分、培养时间等因素对外植体分化的影响，旨在优化遗传转化条件，提高大豆转基因的遗传转化效率。研究结果表明，以健康外植体获得率、抗性丛生芽获得率和抗性芽伸长比率为指标，筛选并建立的优化转化系统为：大豆萌发和不定芽诱导时分别加入0.5 mg L-16-BA和1.0 mg L-16-BA，浸染时间为30 min、共培养时间为3 d；外植体大小为2/3子叶，kan抗性筛选浓度第一阶段和第二阶段分别是60和50 mg L-1；利用上述方法，已获得中豆32转基因再生植株，经PCR分子检测，证明目的基因TaNHX2已导入并整合到大豆基因组中，转化率为3.78%
Abstract:
Transgenic soybeans have been widely planted aboard，which leading the development of the transgenic crop in the world.Establishment of an efficient genetic transformation technology system will facilitate physiological and molecular biology studies as well as the improvement of cultivars in China. The objective of this study was to optimize the conditions of the cotyledon node system of Agrobacterium-mediated soybean transformation so that the transformation plants can be obtained.Four soybean genotypes of Zhongdou 32，Peking，Zaoshu 18 and Suinong 14 was used to introduce the wheat transcription factor Na+/H+antiporter into soybean genome by Agrobacterium-mediated transformation.The proportions of healthy explants，resistance tufty shoots and elongated shoots used as criteria for selection of proper conditions.The results indicated that the optimal system included in 0.5 mg L-1and 1.0 mg L-1of 6-BA for germination medium and shoot induction，2/3 cotyledon infected with 30 min for infection，3 days for co-culture，selection with 60 and 50 mg L-1Kan during the first and second shoot initiation stages. Based on the optimized condition positive transformed plants containing TaNHX2 was obtained，which tested by specific PCR，indicating that Na+/H+ antiporter gene was into the genome of soybean.The results indicated that genotypes mainly affected rates of transformation and regeneration.The rate of genetic transformation in cultivar Zhongdou 32 was 3.78%.

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备注/Memo

基金项目：农作物基因资源与遗传改良国家重大科学工程开放课题（NFCRI 2006）

作者简介：姬月梅（1981-），女，，硕士，主要从事大豆转基因研究。E-mail：ji_yuemei@126.com
通讯作者(Corresponding author)：邱丽娟，研究员，博士。Tel：010-62186623;E-mail: qiu_lijuan@263.net

更新日期/Last Update: 2014-10-08