农杆菌侵染玉米芽尖导入<em>psy</em>基因的研究

中国生物工程杂志

2012, Vol. 32

Issue (08): 36-40

研究报告

农杆菌侵染玉米芽尖导入psy基因的研究

汪婷婷^1,2, 季静², 王罡², 关春峰², 张烈³, 金超²

1. 天津大学化工学院天津 300072;
2. 天津大学遗传工程研究所天津 300072;
3. 天津科润津丰种业有限责任公司天津 300384

Study on Agrobacterium-mediated Transformation of psy Gene into Shoot Meristem of Maize

WANG Ting-ting^1,2, JI Jing², WANG Gang², GUAN Chun-feng², ZHANG Lie³, JIN Chao ²

1. School of Chemical Engineering and Technology,Tianjin University,Tianjin 300072,China;
2. Research Institute of Genetic Engineering,Tianjin University,Tianjin 300072,China;
3. Tianjin Kemel Jinfeng Seed Industry Co. Ltd,Tianjiin 300384,China

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摘要： 以玉米自交系天塔五母、7922的芽尖为受体,用农杆菌介导法将psy基因转入玉米中。以植株的转化率为指标,研究了真空处理方式,真空处理时间及共培养时间对转化率的影响。结果表明,当在真空条件下侵染20min,共培养3天时转化率最高。用200mg/L的草胺膦溶液筛选,获得抗性植株经PCR检测有16株表现阳性,其中天塔五母有12株,7922有4株。RT-PCR及高效液相色谱(HPLC)检测结果表明psy基因已经整合进玉米基因组并能正常转录,转基因玉米中总类胡萝卜素含量比野生型玉米提高了25%。

关键词： 芽尖; 农杆菌介导法; psy基因

Abstract: The psy gene was transferred into the shoot meristems of two different maize inbred lines including maternal Tian tawu and 7922 by Agrobacterium-mediated transformation. Factors influencing transformation efficiency were optimized.The results indicated that the optimal transformation condition is infection time with vacuum at 20 min and co-cultivation time for 3 days.The transgenic plants were obtained after selected by 200mg/L PPT and 16 of them were confirmed positive by PCR amplification. RT-PCR and HPLC analysis results showed that psy gene had been integrated into the maize genome and transcribed normally, the total carotenoid content in transgenic maize was increased 25% compared to the wild type. This method eliminates the tedious process of tissue culture and proves to be a simple, strong operable transformation method.

Key words: Shoot meristems Agrobacterium-mediated transformation psy gene

收稿日期: 2012-03-13 出版日期: 2012-08-25

ZTFLH:

Q943

基金资助: 国家转基因重大专项项目(2009ZX08003-019B)、国家转基因重大专项的重大项目子课题(2008ZX08003-005)、2010年度高等学校博士学科点专项科研基金(20100032110060)资助项目

通讯作者: 季静, 王罡 E-mail: jijingtjdx@163.com; wanggangtjdx@126.com

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引用本文:

汪婷婷, 季静, 王罡, 关春峰, 张烈, 金超. 农杆菌侵染玉米芽尖导入psy基因的研究[J]. 中国生物工程杂志, 2012, 32(08): 36-40.

WANG Ting-ting, JI Jing, WANG Gang, GUAN Chun-feng, ZHANG Lie, JIN Chao. Study on Agrobacterium-mediated Transformation of psy Gene into Shoot Meristem of Maize. China Biotechnology, 2012, 32(08): 36-40.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I08/36

[1] Fromm M E,Morrish F,Armstrong C,et al. Inheritance and expression of chimaeric genes in the progeny of transgenic maize plants. Biotechnology,1990,8:833-839.
[2] Huang X Q, Wei Z M.Successful Agrobacterium-mediated genetic transformation of maize elite inbred lines. Plant Cell,Tissue and Organ Culture,2005,83:187-200.
[3] Cheng M,Fry J E,Pang S,et al.Genetic transformation of wheat mediated by Agrobacterium tumefaciens. Plant Physiol,1997,115:971-980.
[4] 陶俊,张上隆,徐昌杰,等.类胡萝卜素合成的相关基因及其基因工程.生物工程学报,2002,18(3):276-281. Tao J,Zhang S L,Xu C J,et al. Gene and gene engineering of carotenoid biosynthesis. Chinese Journal of Biotechnology,2002,18(3):276-281.
[5] 霍培,季静,王罡,等.植物类胡萝卜素生物合成及功能.中国生物工程杂志,2011,31(11):107-113. Huo P,Ji J,Wang G,et al. Biosynthesis and function of carotenoid in plant. China Biotechnology,2011,31(11):107-113.
[6] 陈中媛,庐山,黄继荣.植物类胡萝卜素代谢与应用.生命科学,2011,23(2):205-211. Chen Z Y,Lu S,Huang J R. Metabolic engineering and applications of plant carotenoid biosynthesis. Chinese Bulletin of Life Sciences,2011,23(2):205-211.
[7] 龚学臣,季静,抗艳红,等.八氢番茄红素合成酶(psy)对大豆的转化.大豆科学,2005,24(1):30-33. Gong X C,Ji J,Kang Y H,et al. Genetic transformation of phytoene synthase gene(PSY)into soybean. Soybean Science,2005, 24(1):30-33.
[8] 于源华,杜柏权,张勇,等.八氢番茄红素合酶基因对人参的遗传转化.中国生物工程杂志,2008, 28(2): 90-95. Yu Y H,Du B Q,Zhang Y,et al. Genetic transformation of phytoene synthase gene in to ginseng callus cells. China Biotechnology,2008, 28(2): 90-95.
[9] 梁广东,卢翠华,王振华,等.农杆菌侵染玉米芽尖转化BcWRKY1转录因子基因的初报. 玉米科学,2011,19(3):56-58. Liang G D,Lu C H,Wang Z H,et al. Introduction of BcWRKY1 gene mediated by Agrobacterium Tumefacien into shoot tip of maize. Journal of Maize Sciences,2011,19(3):56-58.

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