一种水稻真空渗透转化装置及方法的研究

钟巧芳,李维蛟,殷富有,张敦宇,陈玲,陈越,曾民,程在全

中国农学通报. 2019, 35(27): 17-22

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中国农学通报 ›› 2019, Vol. 35 ›› Issue (27) : 17-22. DOI: 10.11924/j.issn.1000-6850.casb18050004
农学 农业基础科学

一种水稻真空渗透转化装置及方法的研究

  • 钟巧芳1, 李维蛟2, 殷富有1, 张敦宇1, 陈玲1, 陈越1, 曾民1, 程在全1
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A Piece of Rice Vacuum Infiltration Equipment and Its Use Method

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摘要

探究适用于水稻的真空渗透转化装置及方法，旨在解决目前水稻转基因效率低、转化体变异多、周期长等问题。以抽穗期的‘云资粳41号’幼穗作为受体，应用自制的水稻真空渗透装置对其短暂抽真空渗透浸染导入外源基因。本试验共计转化水稻30株，收获19860粒T0代种子。研究发现不同浓度除草剂对T0代种子的萌发影响不显著，但对T1代材料的生长发育却具有明显抑制作用，因此选择苗期喷施法进行抗性转化子筛选，最佳筛选浓度为20 mg/L。经除草剂抗性筛选，获得抗性苗135株。经PCR分子检测，最终获得阳性苗114株，表明外源DNA已成功导入到受体水稻基因组中。本研究首次应用真空渗透法快速实现了水稻外源基因的转化，为水稻及其他作物的基因转化提供了一种快速、有效的新装置和新途径。

Abstract

In order to solve the problems of low efficiency, high mutation and long period in rice genetic transformation, this paper first reported a piece of new equipment for vacuum infiltration transformation and its use method in rice. Rice variety ''Yunzijing 41'' was used as receptor in transformation. The whole rice plants at flowering stage were put inside the vacuum infiltration equipment. The young panicles in vivo were dipped in agrobacterium infection buffer. Then the agrobacterium-mediated transformation was carried out and enhanced by short time vacuum infiltration. In this experiment, a total of 30 rice plants were transformed and 19,860 T0 seeds were harvested. It was found that different concentrations of herbicides had no significant effect on the germination of T0 seeds, but had significant inhibitory effects on the growth and development of T1 plantlets. Therefore, the resistant transformants were selected by spraying herbicide at the seedling stage. The optimal screening concentration was 20 Mg/L. 135 resistant rice plants were obtained by herbicide resistance screening. 114 positive rice plants were obtained by PCR confirmation. It indicated that the foreign DNA has been successfully introduced into the recipient rice genome. This study first used the vacuum infiltration method and successfully introduced the exogenous genes into rice. That means we had developed a rapid and effective new way as well as a piece of equipment for gene transformation in rice and other cereal crops.

关键词

水稻；真空渗透；新转化方法；装置

Key words

rice; vacuum infiltration; new transformation method; equipment

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钟巧芳,李维蛟,殷富有,张敦宇,陈玲,陈越,曾民,程在全. 一种水稻真空渗透转化装置及方法的研究. 中国农学通报. 2019, 35(27): 17-22 https://doi.org/10.11924/j.issn.1000-6850.casb18050004
A Piece of Rice Vacuum Infiltration Equipment and Its Use Method. Chinese Agricultural Science Bulletin. 2019, 35(27): 17-22 https://doi.org/10.11924/j.issn.1000-6850.casb18050004

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