一种简便的获得无标记耐盐转基因水稻植株的NaCl有效筛选浓度选择法

doi:10.3969/j.issn.1001-7216.2011.03.003

中国水稻科学

一种简便的获得无标记耐盐转基因水稻植株的NaCl有效筛选浓度选择法

赵艳^1,2，*;罗园园¹;张晓丽¹;郭龙彪²; 钱前^2，*; 董禹然¹

¹浙江工商大学食品与生物工程学院, 浙江杭州 310012; ²中国水稻研究所水稻生物学国家重点实验室/国家水稻改良中心, 浙江杭州 310006; *通讯联系人, E-mail: yanzhao9918@163.com; qianqian188@hotmail.com

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-05-10 发布日期:2011-05-10

A Simple Selection Method for Effective Screening Concentration of NaCl for Generating Marker-Free Salt-Tolerant Transgenic Rice

ZHAO Yan^1,2,*; LUO Yuan-yuan¹; ZHANG Xiao-li¹; GUO Long-biao²; QIAN Qian2，*; DONG Yu-ran¹

¹College of Food Science, Biotechnology Engineering, Zhejiang Gongshang University, Hangzhou 310012, China; ² State Key Laboratory of Rice Biology/Chinese National Center for Rice Improvement, China National Rice Research Institute, Hangzhou 310006, China; *Corresponding authors, E-mail：yanzhao9918@163.com; qianqian188@hotmail.com

Received:1900-01-01 Revised:1900-01-01 Online:2011-05-10 Published:2011-05-10

摘要/Abstract

摘要： 以4个粳稻品种(日本晴、秀水11、武运粳7号、中花11)为材料, 研究发现，在含NaCl的NB培养基上，NaCl对水稻种子发芽的抑制率与对成熟胚愈伤组织生长的抑制率之间呈线性正相关。水稻种子发芽抑制率达50%左右时的NaCl浓度与愈伤组织生长抑制率达80%左右时的NaCl浓度一致，可以作为耐盐基因转化水稻时NaCl有效筛选浓度的重要参考。由此获得日本晴、武运粳7号和中花11 NaCl有效筛选浓度为200 mmol/L,秀水11 NaCl有效筛选浓度为250 mmol/L。采用上述浓度的NaCl对BADH基因转化的日本晴和秀水11愈伤组织进行筛选，成功获得了无标记基因的转基因水稻植株，转化率分别达到38.9%和32.0%。建立的NaCl筛选浓度确定法通过测定NaCl对水稻种子的发芽抑制率来确定水稻耐盐基因转化时合适的NaCl筛选浓度，操作简便，实验周期短，无需其他抗性标记基因。

关键词: 水稻, 耐盐基因, 甜菜碱醛脱氢酶基因, 转化, 无标记基因

Abstract: The relevance between inhibition rates of rice seed germination and mature embryo callus growth by NaCl at different concentrations was studied by using four japonica rice cultivars (Nipponbare, Xiushui 11, Wuyunjing 7 and Zhonghua 11) as materials. Results showed that there was a significant positive correlation between the inhibition rates of rice seed germination and mature embryo callus growth under salt stress (on NB medium containting NaCl). The NaCl concentration at which the inhibition rate of rice seed germination reached about 50% was concordant with the concentration at which the inhibition rate of mature embryo callus growth reached about 80%. These could be used as important references to determine the effective concentrations of NaCl for screening of transformed cells in rice transformation with salttolerant gene. The effective NaCl concentrations were 200 mmol/L for Nipponbare, Wuyunjing 7 and Zhonghua 11 and 250 mmol/L for Xiushui 11. Transgenic rice plants were successfully obtained with the NaCl concentration as screening standard in the transformation of Nipponbare and Xiushui 11 with salttolerant gene BADH. The transformation frequency of Nipponbare and Xiushui 11 were 38.9% and 320%, respectively. The method that the suitable concentration of NaCl for screening in rice transformation was determined by testing the inhibition rate of rice seed germination, it is free of resistance marker genes and easy to operate with short experimental duration.

Key words: rice, salttolerance gene, betaine aldehyde dehydrogenase gene, transformation, marker-free

赵艳,罗园园,张晓丽,郭龙彪, 钱前, 董禹然. 一种简便的获得无标记耐盐转基因水稻植株的NaCl有效筛选浓度选择法[J]. 中国水稻科学 2011,25(3): 243-248 . , DOI: 10.3969/j.issn.1001-7216.2011.03.003 .

ZHAO Yan,LUO Yuan-yuan,ZHANG Xiao-li,GUO Long-biao,QIAN Qian,DONG Yu-ran. A Simple Selection Method for Effective Screening Concentration of NaCl for Generating Marker-Free Salt-Tolerant Transgenic Rice[J]. Chinese Journal of Rice Science 2011,25(3): 243-248 ., DOI: 10.3969/j.issn.1001-7216.2011.03.003 .

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一种简便的获得无标记耐盐转基因水稻植株的NaCl有效筛选浓度选择法

A Simple Selection Method for Effective Screening Concentration of NaCl for Generating Marker-Free Salt-Tolerant Transgenic Rice

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