虎杖白藜芦醇合酶基因转化拟南芥的研究

doi:10.3969/j.issn.1000-565X.2011.05.024

华南理工大学学报（自然科学版） ›› 2011, Vol. 39 ›› Issue (5): 138-142.doi: 10.3969/j.issn.1000-565X.2011.05.024

虎杖白藜芦醇合酶基因转化拟南芥的研究

柳忠玉¹ 庄楚雄² 生书晶¹ 邵利¹ 赵树进³

1．华南理工大学生物科学与工程学院，广东广州 510006；2．华南农业大学生命科学学院，广东广州 510642；3．广州军区广州总医院，广东广州 510010

收稿日期:2010-10-12 修回日期:2010-12-29 出版日期:2011-05-25 发布日期:2011-04-01
通信作者: 赵树进(1958-)，男，教授，博士生导师，主要从事生物制药研究 E-mail:gzzsjzhs@163.com
作者简介:柳忠玉(1978-)，女，博士生，主要从事药用植物次生代谢研究
基金资助:
广东省自然科学基金资助项目(10151001002000012)；广东省科技计划项目(2010B060200009

Transformation of Polygonum cuspidatum Resveratrol Synthase Gene into Arabidopsis thaliana

Liu Zhong-yu¹ Zhuang Chu-xiong² Sheng Shu-jing¹ Shao Li¹ Zhao Shu-jin³

1．School of Biological Science and Engineering，South China University of Technology，Guangzhou 510006，Guangdong，China；2．College of Life Science，South China Agricultural University，Guangzhou 510642，Guangdong，China；3．General Hospital of Guangzhou Military Command，Guangzhou 510010，Guangdong，China

Received:2010-10-12 Revised:2010-12-29 Online:2011-05-25 Published:2011-04-01
Contact: 赵树进(1958-)，男，教授，博士生导师，主要从事生物制药研究 E-mail:gzzsjzhs@163.com
About author:柳忠玉(1978-)，女，博士生，主要从事药用植物次生代谢研究
Supported by:
广东省自然科学基金资助项目(10151001002000012)；广东省科技计划项目(2010B060200009

摘要/Abstract

摘要： 为验证虎杖白藜芦醇合酶基因PcRS在转基因植物中合成白藜芦醇的有效性，构建了植物表达载体pCAM1380-35S-PcRS，通过农杆菌介导，利用花序浸泡法转化拟南芥。转化子在含有潮霉素的培养基上经筛选后，利用PCR和Southern杂交技术检测，进一步证实PcRS基因已整合到拟南芥基因组。经过选育得到了遗传稳定的T3代纯合子转基因拟南芥株系。Northern杂交证实外源基因在转基因拟南芥纯合子株系中实现表达，并且通过HPLC法检测到终产物是反式白藜芦醇苷。两周大拟南芥幼苗产生的反式白藜芦醇苷为136μg/g（鲜重），干重为1813μg/g。

关键词: 虎杖, 白藜芦醇合酶基因, 拟南芥, 白藜芦醇, 白藜芦醇苷

Abstract:

In order to reveal the effectiveness of Polygonum cuspidatum resveratrol synthase（PcRS） gene in improving the resveratrol accumulation in foreign species,first,the plant expression vector pCAM1380-35S-PcRS was constructed and introduced into Arabidopsis thaliana by Agrobacterium tumefaciens by means of the floral dip me-thod.Next,the transformants were screened on the medium containing hygromycin.Then,the integration of PcRS gene into the transgenic plants was verified via the PCR and the Southern blotting.Moreover,independent homozygous transgenic Arabidopsis thaliana lines with genetical stability were obtained after the selection of T3 progenies,and the expression of the gene transferred into Arabidopsis thaliana was confirmed by Northern blotting.Finally,an end product,namely trans-piceid,was identified by HPLC,the content of which in 2-week-old plants being 136 μg/g（fresh weight） or 1 813 μg/g（dry weight）.

Key words: Polygonum cuspidatum, resveratrol synthase gene, Arabidopsis, resveratrol, piceid

柳忠玉庄楚雄生书晶邵利赵树进. 虎杖白藜芦醇合酶基因转化拟南芥的研究[J]. 华南理工大学学报（自然科学版）, 2011, 39(5): 138-142.

Liu Zhong-yu Zhuang Chu-xiong Sheng Shu-jing Shao Li Zhao Shu-jin. Transformation of Polygonum cuspidatum Resveratrol Synthase Gene into Arabidopsis thaliana[J]. Journal of South China University of Technology (Natural Science Edition), 2011, 39(5): 138-142.

[1]	李琳, 余安玲, 吴轶, 等. 改性果胶与白藜芦醇的相互作用机制及抗氧化能力[J]. 华南理工大学学报（自然科学版）, 2021, 49(9): 1-10.
[2]	叶勇邢海婷郭亚. 白藜芦醇二聚体与眼镜蛇神经毒素的作用机制[J]. 华南理工大学学报（自然科学版）, 2014, 42(2): 21-26,32.

虎杖白藜芦醇合酶基因转化拟南芥的研究

Transformation of Polygonum cuspidatum Resveratrol Synthase Gene into Arabidopsis thaliana

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