<FONT face="Verdana">A Simple Selection Method for  Effective Screening Concentration of NaCl for Generating Marker-Free Salt-Tolerant Transgenic Rice</FONT>

ZHAO Yan,LUO Yuan-yuan,ZHANG Xiao-li,GUO Long-biao,QIAN Qian,DONG Yu-ran

doi:10.3969/j.issn.1001-7216.2011.03.003

2011 , Vol. 25 >Issue 3: 243 - 248

DOI: https://doi.org/10.3969/j.issn.1001-7216.2011.03.003

研究报告

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

Expand

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 date: 1900-01-01

Revised date: 1900-01-01

Online published: 2011-05-10

Fold

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

Cite this article

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

References

[1]张鹤山, 刘洋, 田宏, 等. 黑麦种子在不同盐分胁迫下萌发特性的研究. 种子, 2009, 28(3): 14-17.

[2]Rai M, He C, Wu R. Comparative functional analysis of three abiotic stress-inducible promoters in transgenic rice. Transgenic Res, 2009, 18: 787-799.

[3]胡婷婷, 刘超, 王健康, 等. 水稻耐盐基因遗传及耐盐育种研究. 分子植物育种, 2009, 7(1): 110-116.

[4]Wang Y, Xue Y, Li J. Towards molecular breeding and improvement of rice in China. Trends Plant Sci, 2005, 10(12): 610-614.

[5]Darbani B, Eimanifar A, Stewart C N, et al. Methods to produce marker-free transgenic plants. Biotechnol J, 2007, 2: 83-90.

[6]Zhu Z, Wu R. Regeneration of transgenic rice plants using high salt for selection without the need for antibiotics or herbicides. Plant Sci, 2008, 174: 519-523.

[7]祁栋灵, 韩龙植, 张三元. 水稻耐盐/碱性鉴定评价方法. 植物遗传资源学报, 2005, 6(2): 226-231.

[8]方先文, 汤陵华, 王艳平. 耐盐水稻种质资源的筛选. 植物遗传资源学报, 2004, 5(3): 295-298.

[9]张燕红, 袁杰, 王奉斌. 不同新疆水稻品种种子发芽耐盐性的比较. 安徽农学通报, 2009, 15(3): 102-103.

[10]Hakim M A, Uraimi A, Begum S M, et al. Effect of salt stress on germination and early seedling growth of rice (Oryza sativa L.). Afr J Biotechnol, 2010, 9(13): 1911-1918.

[11]Jamil M, Rha E S. Response of transgenic rice at germination and early seedling growth under salt stress. Pak J Biol Sci, 2007, 10(23): 4303-4306.

[12]卢德赵, 王慧中, 华志华, 等. 转甜菜碱醛脱氢酶基因水稻的获得及其耐盐性研究. 科技通报, 2003, 19(3): 179-182.

[13]Edwards K, Johnstone C, Thompson C. A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucl Acids Res, 1991, 19: 1349.

[14]卢德赵, 祝水金, 钱前, 等. BADH基因转化水稻的方法比较. 中国水稻科学, 2003, 17(4): 323-327.

[15]林秀峰, 刘志铭. 转甜菜碱醛脱氢酶基因水稻的获得. 吉林农业科学, 2005, 30(2): 7-9.

Options

Outlines

模态框（Modal）标题

Abstract

Cite this article

References