园艺学报2024,Vol.51Issue(10) :2439-2448.DOI:10.16420/j.issn.0513-353x.2023-0944

发根农杆菌介导的菜用豌豆遗传转化体系构建

Establishment of Agrobacterium rhizogenes-Mediated Genetic Transformation System in Vegetable Pea

冯志娟 刘娜 卜远鹏 张古文 王斌 龚亚明
园艺学报2024,Vol.51Issue(10) :2439-2448.DOI:10.16420/j.issn.0513-353x.2023-0944

发根农杆菌介导的菜用豌豆遗传转化体系构建

Establishment of Agrobacterium rhizogenes-Mediated Genetic Transformation System in Vegetable Pea

冯志娟 1刘娜 1卜远鹏 1张古文 1王斌 1龚亚明1
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作者信息

  • 1. 农业农村部南方豆类蔬菜种质创新与分子育种重点实验室(部省共建),省部共建农产品质量安全危害因子与风险防控国家重点实验室,浙江省农业科学院蔬菜研究所,杭州 310000
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摘要

创建了一套由发根农杆菌介导,简单高效的菜用豌豆遗传转化体系.以播种于无菌基质中菜用豌豆'浙豌1号'为材料,以在子叶节和下胚轴连接处切割后的上部幼苗为外植体,用导入pCAMBIA3301-GUS载体的发根农杆菌AR1193诱导外植体生成毛状根,毛状根诱导率为100%.7 d苗龄幼苗更有利于毛状根的诱导,平均生根数为14.60.对毛状根进行GUS基因RT-PCR检测及GUS染色验证,发现外源GUS基因可在毛状根中成功表达,毛状根转化率为35.21%.

Abstract

This study established a simple and efficient Agrobacterium rhizogenes-mediated genetic transformation system in vegetable pea.The vegetable pea seeds of'Zhewanl'were sown into pots with wet sterile vermiculite.The hypocotyls of vegetable pea seedlings were cut at the junction of cotyledon node and hypocotyl and the residual seedlings were selected as explants.The hairy roots were induced with Agrobacterium rhizogenes AR1 193 strain harboring the construct of pCAMBIA3301-GUS.The results indicated that 100%of the inoculated explants could produce hair roots.The effect of different seedling age of explant on hairy root induction was also investigated.It was found that the 7-day-old seedling was favorable for hair root induction and the number of hairy root per seedling was 14.60.Further,the hair roots were amplified by RT-PCR of GUS gene and stained with the GUS dye.It was proven that the exogenous GUS gene could be successfully expressed in the hair roots and 35.21%of the hair roots were transgenic positive roots.

关键词

菜用豌豆/发根农杆菌/苗龄/毛状根/遗传转化/转基因根

Key words

vegetable pea/Agrobacterium rhizogenes/seeding age/hairy root/genetic transformation/transgenic root

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基金项目

浙江省公益项目(LGN21C150007)

浙江省蔬菜新品种选育重大科技专项(2021C02065-6)

国家自然科学基金项目(31601767)

浙江省重点研发计划项目(2022C02016)

浙江省重点研发计划项目(2024C04003)

出版年

2024
园艺学报
中国园艺学会 中国农业科学院蔬菜花卉研究所

园艺学报

CSTPCD北大核心
影响因子:1.127
ISSN:0513-353X
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