水稻无选择标记Xa21转基因系CX8621的获得与遗传分析
# 共同第一作者;
收稿日期: 2015-09-07
修回日期: 2015-10-16
网络出版日期: 2016-01-10
基金资助
国家转基因生物新品种培育重大专项(2008/2014ZX08001-002);中国科学院知识创新方向项目(KSCX2-EWN-01)
Generation and Genetic Analysis of Xa21 Transgenic Rice Line CX8621 Without Selectable Markers
# These authors contributed equally to this work;
Received date: 2015-09-07
Revised date: 2015-10-16
Online published: 2016-01-10
夏志辉, 刘鹏程, 高利芬, 刘栋峰, 姜丽, 江光怀, 郭乐群, 翟文学 . 水稻无选择标记Xa21转基因系CX8621的获得与遗传分析[J]. 中国水稻科学, 2016 , 30(1) : 10 -16 . DOI: 10.16819/j.1001-7216.2016.5130
Selectable marker-free and stable inheritance are the basic requirements for safe application of transgenic crops. Following the regulations approved by the National Committee for Transgenic Biosafety, the wide spectrum bacterial blight resistance gene Xa21 was transferred into a widely used restorer line Minghui 86 by using a double right-borders T-DNA vector through Agrobacterium tumefaciens-mediated system. The transgene Xa21 was confirmed by resistance identification, PCR analysis and Southern blot in the progenies. The single-copy homozygous transgenic line CX8621 without selectable marker and vector backbone sequences was obtained from T3 generation. The transgenic restorer line CX8621 has stable bacterial blight resistance with years of observation and identification. Up to now, CX8621 has self-fertilized to 16th generation with stable inheritance of transgene Xa21 and passed the bio-safety evaluation of small and large scale of field tests (environmental release and productive test).
Key words： transgenic rice; bacterial blight; Xa21; selectable maker-free; stable inheritance
| [1] | Lu B R, Snow A A.Gene flow from genetically modified rice and its environmental consequences.BioScience, 2005, 55(8): 669-678. |
| [2] | 程式华,胡培松.中国水稻科技发展战略.中国水稻科学,2008,22(3):223-226. |
| [2] | Chen S Y, Hu P S.Development strategy of rice science and technology in China.Chin J Rice Sci, 2008,22(3):223-226. (in Chinese with English abstract) |
| [3] | 陈浩,林拥军,张启发. 转基因水稻研究的回顾与展望.科学通报,2009,54(18):2699-2717. |
| [3] | Chen H, Lin Y J, Zhang Q F.Review and prospect of transgenic rice research.Chin Sci Bull, 2009, 54: 2699-2717. (in Chinese with English abstract) |
| [4] | Tang W, Chen H,Xu C, et al.Development of insect-resistant transgenic indica rice with a synthetic cry1C* gene.Mol Breeding, 2006, 18(1): 1-10. |
| [5] | 尚玉花,李莉,陆徐忠,等.利用Cre/loxP系统删除转Bt基因水稻中的选择标记基因.生物学杂志,2011,28(2):5-8. |
| [5] | Shang Y H, Li L, Lu X Z, et al.Using Cre/loxP system to excise marker gene in Bt-transgenic rice.J Biol, 2011, 28(2): 5-8. (in Chinese with English abstract) |
| [6] | 夏志辉,李晓兵,陈彩艳,等.无选择标记和载体骨干序列的Xa21转基因水稻的获得. 生物工程学报,2006,22(2):204-210. |
| [6] | Xia Z H, Li X B, Chen C Y, et al.Generation of selectable marker-free and vector backbone sequence-free Xa21 transgenic rice.Chin J Biotech, 2006, 22(2): 204-210. (in Chinese with English abstract) |
| [7] | 吴爱忠,唐克轩,潘俊松,等.转基因培育抗除草剂水稻.遗传学报,2000,27(11):992-998. |
| [7] | Wu A Z, Tang K X, Pan J S, et al.Production of herbicide-resistant rice with transforming heterogene.Acta Gene Sin, 2000, 27(11): 992-998. (in Chinese with English abstract) |
| [8] | 于恒秀,刘巧泉,陈秀花,等.根癌农杆菌介导的水稻转化体系的优化及转反义Wx基因植株的获得.中国水稻科学,2002, 16(4):304-310. |
| [8] | Yu H X, Liu Q Q, Chen X H, et al.Optimization of Agrobacterium-mediated transformation of rice and regeneration of transgenic plants with antisense Wx gene.Chin J Rice Sci, 2002, 16(4): 304-310. (in Chinese with English abstract) |
| [9] | Gao Y F, Jing Y X, Shen S H, et al.Transfer of lysine-rich protein gene into rice and production of fertile transgenic plants.Acta Bot Sin, 2001, 43(5): 506-511. |
| [10] | 朱祯. 转基因水稻研发进展.中国农业科技导报,2010,12(2):9-16. |
| [10] | Zhu Z.Progress in research and development of transgenic rice.J Agric Sci Tech, 2010, 12(2): 9-16. (in Chinese with English abstract) |
| [11] | 李黎红,叶卫军,郭龙彪.我国转基因水稻研究进展和商业化前景分析.中国稻米,2012,18(6):1-4. |
| [11] | Li L H, Ye W J, Guo L B.Research progress and commercialization prospect of transgenic rice in China.China Rice, 2012, 18(6): 1-4. (in Chinese with English abstract) |
| [12] | 农业部. 农业转基因生物安全评价管理办法.中华人民共和国国务院公报,2002,35:28-33. |
| [12] | China Ministry of Agriculture. Measures for the safety assessment of agricultural genetically modified organisms. Gazette of the State Council of the People's Republic of China, 2002, 35: 28-33. (in Chinese) |
| [13] | Maqbool S B, Christou P.Multiple traits of agronomic importance in transgenic indica rice plants: Analysis of transgene integration patterns, expression levels and stability.Mol Breeding, 1999, 5(5): 471-480. |
| [14] | Song W Y, Wang G L, Chen L L, et al.A receptor kinase-like protein encoded by the rice disease resistance gene, Xa21.Science, 1995, 270(5243): 1804-1806. |
| [15] | Gao L F, Xia Z H, Jiang G H, et al.Generation of marker-free, bacterial blight-resistant transgenic sterile line and hybrid rice with Xa21.Plant Breeding, 2011, 130(4):438-443. |
| [16] | 翟文学, 周永力. 由农杆菌介导将白叶枯病抗性基因 Xa21 转入我国的5个水稻品种. 中国科学: C辑,2000,30(2):200-206. |
| [16] | Zhai W X, Zhou Y L.The rice bacterial blight resistance gene Xa21 was transferred into 5 rice varieties in China. Sci China:Series C, 2000, 30(2): 200-206. (in Chinese with English abstract) |
| [17] | Wang G L, Song W Y, Ruan D L, et al.The cloned gene, Xa21, confers resistance to multiple Xanthomonas oryzae pv. oryzae isolates in transgenic plants.Mol Plant Microbe In, 1996, 9(9): 850-855. |
| [18] | 武小霞,张彬彬,王志坤,等.转基因作物的生物安全性管理及安全评价.作物杂志,2010,4(1):1-4. |
| [18] | Wu X X, Zhang B B, Wang Z K, et al.Safety management and assessment of genetically modified organisms.Crops, 2010, 4(1): 1-4. (in Chinese with English abstract) |
| [19] | 何礼健,周玉婷,左停.转基因生物技术在农业领域的发展现状分析.安徽农业科学,2011,39(1):66-68. |
| [19] | He L J, Zhou Y T, Zuo T.Analysis on the development status of transgenic biotechnology in agricultural field.J Anhui Agri Sci, 2011, 39(1): 66-68. (in Chinese with English abstract) |
| [20] | 陈松彪, 李旭刚, 王锋, 等. 无选择标记转基因植物的培育. 中国生物工程杂志, 2005,25(2): 1-7. |
| [20] | Chen S B, Li X G, Wang F, et al.Culturing maker-free transgenic plants.China Biotechnol, 2005, 35(2): 1-7. (in Chinese with English abstract) |
| [21] | 于恒秀,刘巧泉,徐丽,等.无抗性选择标记转基因软米和糯稻新品系的选育及中间试验.作物学报,2009,35(6):967-973. |
| [21] | Yu H X, Liu Q Q, Xu L, et al.Breeding and field performance of novel soft and waxy transgenic rice lines without selectable markers.Acta Agron Sin, 2009, 35(6): 967-973. (in Chinese with English abstract) |
| [22] | Yoder J I, Goldsbrough A P.Transformation systems for generating marker-free transgenic plants.Biotechnology, 1994, 12(3): 263-267. |
| [23] | 李晓飞,谭小力,李俊,等.转基因水稻检测用阳性质粒分子的构建及应用.生物技术通报,2013,7:71-77. |
| [23] | Li X F, Tan X L, Li J, et al.Construction and application of a plasmid reference molecule for detecting transgenic rice.Biotechnol Bull, 2013, 7: 71-77. (in Chinese with English abstract) |
| [24] | 金万梅,潘青华,尹淑萍,等.外源基因在转基因植物中的遗传稳定性及其转育研究进展.分子植物育种,2005,3(6):112-116. |
| [24] | Jin W M, Pan Q H, Yin S P, et al.Progress of the genetic stability and breeding behavior of foreign gene in genetically modified plants.Mol Plant Breeding, 2005, 3(6): 112-116. (in Chinese with English abstract) |
/
| 〈 |
|
〉 |