番茄红素β-环化酶基因的玉米转化及 后代遗传分析

中国生物工程杂志

2012, Vol. 32

Issue (07): 43-48

研究报告

番茄红素β-环化酶基因的玉米转化及后代遗传分析

霍培, 季静, 王罡, 关春峰, 金超

天津大学农业与生物工程学院天津 300072

Transformation and Genetic Analysis of Maize with the Lycb

HUO Pei, JI Jing, WANG Gang, GUAN Chun-feng, JIN Chao

School of Agriculture and Bioengineering, Tianjin University, Tianjin 300072, China

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摘要： 利用农杆菌介导法将番茄红素β-环化酶基因(Lycb)转入由玉米自交系天塔五号植株,分析基因在T₀转化及后代的遗传情况,结果表明,在27株T₀转基因植株中, PCR初步检测后8株呈阳性;将T₁代转基因植株以株系为单位用200mg/L草铵膦抗性筛选后,收获抗性植株种子。T₂代转基因植株进一步进行PCR、RT-PCR和田间草铵膦涂抹检测,结果表明,PCR、RT-PCR为阳性的6个株系植株均具有草铵膦抗性。选取6株阳性植株提取叶片总类胡萝卜素,经HPLC分析其β-胡萝卜素含量显著高于野生型,表明目的基因Lycb成功的转入玉米,并得到了稳定遗传。

关键词： 番茄红素β-环化酶基因; Bar基因; 玉米; 遗传转化; HPLC

Abstract: Lycb gene was transformed into embryonic callus derived from Tianta Fifth inbred lines by Agrobacterium-mediated transformation, and the independent T₀ transformants and their progenies genetics were analyzed. PCR analysis with 27 T₀ transformed plants testified 8 positive plantlets. T₁ plantlets were selected by 200 mg/L PPT and seeds of the resistance plants were harvested. T₂ transgenic plants were analyzed using PCR, RT-PCR, and the results showed that the six positive lines detected were further confirmed to have PPT resistance. HPLC analysis showed the total beta carotene content from transformed leaves was significantly higher than that of the wild type. The results show that the Lycb gene was able to inherit in the T₂ progeny, and transcribed and translated effectively.

Key words: Lycopene β-cyclase(Lycb) Bar Maize Transformation HPLC

收稿日期: 2012-03-12 出版日期: 2012-07-25

ZTFLH:

Q78

基金资助: 国家转基因生物新品种培育科技重大专项资助项目(2009ZX08003-019B;2011ZX08003-005;2011ZX08004-001;2009ZX08010-013B)

通讯作者: 季静 E-mail: jijingtjdx@163.com

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引用本文:

霍培, 季静, 王罡, 关春峰, 金超. 番茄红素β-环化酶基因的玉米转化及后代遗传分析[J]. 中国生物工程杂志, 2012, 32(07): 43-48.

HUO Pei, JI Jing, WANG Gang, GUAN Chun-feng, JIN Chao. Transformation and Genetic Analysis of Maize with the Lycb. China Biotechnology, 2012, 32(07): 43-48.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I07/43

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