中国水稻科学 ›› 2013, Vol. 27 ›› Issue (5): 447-456.DOI: 10.3969/j.issn.1001-7216.2013.05.001

• 研究报告 •    下一篇

水稻CYP81A6基因干扰对其他基因表达的影响

杨阳1,刘金华1,高其康1,2,*   

  1. 1浙江大学 农业与生物技术学院 昆虫科学研究所,  杭州 310058; 2浙江大学 农生环测试中心, 杭州 310058;
  • 收稿日期:2013-05-15 修回日期:2013-08-05 出版日期:2013-09-10 发布日期:2013-09-10
  • 通讯作者: 高其康1,2,*
  • 基金资助:

    浙江省自然科学基金重点资助项目(Z3080241)。

Knockdown of CYP81A6  Gene  Affects  Expression of Other Genes   in Rice

YANG Yang1, LIU   Jinhua1, GAO Qikang 1,2,*   

  1. 1 Institute of Insect Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China;  2 Analysis Center of Agrobiology and Environmental Sciences, Zhejiang University, Hangzhou 310058, China;
  • Received:2013-05-15 Revised:2013-08-05 Online:2013-09-10 Published:2013-09-10
  • Contact: GAO Qikang1,2,*

摘要: 为了明确抑制基因CYP81A6的表达对水稻的自身功能基因表达的影响,对亲本水稻秀水110和转基因水稻4503的mRNA进行深度测序,并将获得的序列信息通过水稻基因数据库和生物信息学手段进行注释和功能基因比较。结果表明,在3529个差异表达的基因中,上调表达基因1018个,下调表达基因2511个;秀水110与4503中共同表达的基因有2533个,仅在4503中表达的基因有363个,仅在秀水110表达的基因有633个。与水稻基因数据库功能已知的296个抗性基因对比,发现4503中差异表达的基因有64个功能已知且与水稻抗性相关,包括2个抗虫相关基因,17个抗病相关基因和45个抗逆相关基因,预示着基因CYP81A6的抑制表达对水稻抗性会产生一定的影响。GO功能和通路显著性富集分析显示,差异表达基因显著富集在38个GO条目和13个通路上,在结合能力(GO: 0020037)和光合作用(KO 00195)通路上富集最为显著。通过对转基因水稻的转录组分析,阐明转基因水稻基因与亲本相比发生的变化,进而分析这些变化对转基因水稻产生的影响,从基因生态学角度为转基因水稻的安全性评价提供科学依据,可作为一种新的方法应用于转基因生物的安全性评价。

关键词: 转基因水稻, CYP81A6, 高通量测序, 差异表达基因, 抗性基因

Abstract: To study the effects of knockdown the gene CYP81A6 on gene expression of transgenic rice,   we report the transcriptome profiling analysis of transgenic rice  4503 and normal control rice Xiushui 110. In this study, mRNA of transgenic rice and its parent (control) were deeply sequenced, sequences information obtained and functional genes comparison against the rice gene database were analyzed. Results showed that 3529 genes in  total were significantly differential expressed, among which 1018 genes were upregulated and 2511 genes were downregulated. The differential expressed genes distribution of the two samples showed that there were 2533 genes expressed in both Xiushui 110 and 4503, 633 genes only expressed in Xiushui 110, while 363 genes expressed just in 4503. Comparative analysis with 296 resistance genes in the rice database showed that 64 resistance genes were involved in rice resistance, including two  insectresistant genes, 17 diseaseresistant genes and 45 stressresistant genes, indicating that resistance of transgenic plant 4503 would be affected. Gene enrichment studies and further analysis showed that differential expressed genes were significantly enriched in 38 GO terms and 13 KO terms in 4503, ‘heme binding’ (GO:0020037) and ‘photosynthesis’ (KO 00195) were enriched most dramatically. Comparative transcriptome analysis of the transgenic rice 4503 and Xiushui 110 was conducted to clarify the changes of gene expression level in 4503. Knockdown of gene CYP81A6 may result in extensive and variable change in the transcriptome of 4503. Viewed from the gene evology point,these observations may provide valuable information to address the safety of genetically modified crops, and it could be used as a new method for  the safety evaluation of genetically modified organisms.

Key words: transgenic rice, CYP81A6, highthroughput sequencing, differential expressed gene, resistance gene

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