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中国农业科学 ›› 2009, Vol. 42 ›› Issue (9): 3334-3338 .doi: 10.3864/j.issn.0578-1752.2009.09.041

• 研究简报 • 上一篇    下一篇

基因pf40在玉米中的遗传转化

李博,于静娟,赵倩,朱登云,敖光明   

  1. (中国农业大学生物学院/农业生物技术国家重点实验室)
  • 收稿日期:2008-11-28 修回日期:2009-03-30 出版日期:2009-09-10 发布日期:2009-09-10
  • 通讯作者: 朱登云

Transformation of the Gene pf40 in Maize#br#

LI Bo, YU Jing-juan, ZHAO Qian, ZHU Deng-yun, AO Guang-ming#br#   

  1. (中国农业大学生物学院/农业生物技术国家重点实验室)
  • Received:2008-11-28 Revised:2009-03-30 Online:2009-09-10 Published:2009-09-10
  • Contact: ZHU Deng-yun

摘要:

【目的】基因pf40来源于谷子未成熟种子cDNA文库,它或其同源基因普遍存在于禾本科植物中。初步研究显示pf40具有促进分蘖(枝)的功能。本文旨在研究玉米的分蘖特性是否与pf40功能相关。【方法】以玉米自交系Z3、Z31与Q31杂交组合的胚性愈伤组织为转化受体,用基因枪轰击法将pf40超表达载体(PF40HS)和抑制表达载体(PF40R)导入玉米。然后观察pf40超表达和抑制表达两种不同表达类型的转基因玉米分蘖表型。【结果】经潮霉素筛选和PCR 检测,共获得53 株T0代转基因玉米。通过对T1、T2代转基因玉米的PCR检测以及Southern 和tRNA dot blot分析表明,外源pf40已整合到玉米基因组中并得到表达,而且能稳定地遗传给下一代;通过对转pf40玉米茎基部的分蘖表型的观察发现,无论是转超表达载体还是抑制表达载体的转基因植株与未转基因植株均无明显差异。【结论】单独调控玉米中的pf40或其同源基因并不能改变玉米的分蘖表型特征,玉米中的pf40或其同源基因的确切功能,尚待进一步研究。

关键词: pf40')">pf40, 玉米, 转基因植株, 分蘖(枝)

Abstract:

【Objective】 Isolated from the cDNA library of millet immature seed, the gene pf40 (or its homolog ) is ubiquitously distributed in cereal’s crop genome. Preliminary studies indicate that the gene pf40 plays a role in promoting tillering (branching). This study aimed to clarify whether the function of gene pf40 (or its homolog) is related to the tillering in maize. 【Method】 Two plant expression vectors with sense and knock-out (RNAi) gene pf40 promoted by 35S promoter were introduced into the embryogenic callus of maize hybrids(Z3, Z31, Q31) through particle bombardment. Regenerated plants were verified to be transgenic by PCR amplification, Southern blot and tRNA dot blot. The observation about the phenotype of the tillers on the base of the transgenic plants stems could determine the effect of the gene pf40 on the tillers. 【Result】 After the hygromycin B selection and PCR screening, 53 transgenic plants were obtained. According to the results of PCR test, Southern blot and tRNA dot blot on T1 and T2 generation, the foreign gene pf40 had been integrated into the maize genome, also it could express and pass generation to generation. After investigated into the phenotypes of both the tiller nodes on the base of stems, unfortunately, no distinct difference was observed between the transgenic plants and the untransgenic plants. 【Conclusion】 The phenotype of the maize without tillers will not be changed by merely altering the expression of the gene pf40 or its homolog. And the exact functions of the gene pf40 or its homolog in maize need further to be investigated.

Key words: pf40')">pf40, maize, transgenic plants, tillering (branching)