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作物学报 ›› 2009, Vol. 35 ›› Issue (10): 1798-1805.doi: 10.3724/SP.J.1006.2009.01798

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦成熟胚脱分化过程中生长素相关基因的表达分析

陈军营，马平安，赵一丹，朱雪萍，崔琰，张艳敏，陈新建*   

  1. 河南农业大学农学院，河南郑州450002
  • 收稿日期:2009-03-25 修回日期:2009-06-25 出版日期:2009-10-12 网络出版日期:2009-08-07
  • 通讯作者: 陈新建, E-mail: xinjian@371.net; Tel: 0371-63558722
  • 基金资助:

    本研究由国家转基因植物研究与产业化专项基金（JY03-B19-2）和河南省杰出人才创新基金（022100090）资助。

Expression of Auxin-Related Genes during Dedifferentiation of Mature Embryo in Wheat

CHEN Jun-Ying,MA Ping-An,ZHAO Yi-Dan,ZHU Xue-Ping,CUI Yan,ZHANG Yan-Min,CHAN Xin-Jian*   

  1. College of Agronomy,Henan Agricultural University,Zhengzhou 450002,China
  • Received:2009-03-25 Revised:2009-06-25 Published:2009-10-12 Published online:2009-08-07
  • Contact: CHEN Jian-Xin,E-mail: xinjian@371.net; Tel: 0371-63558722

PDF

2036

被引次数

19

摘要：

利用Affymetrix小麦基因芯片研究了小麦成熟胚在MS+2,4-D (2 mg L-1)培养基上脱分化过程种基因表达变化，用NCBI、DATF和DRTF等生物信息学相关网站对基因表达信息进行处理，并针对生长素相关基因的变化情况进行分析，结果表明，有80个生长素相关基因在小麦成熟胚脱分化过程中的2、6、12、24和72 h等不同时间点，至少发生了一次有意义的表达变化，其中41个在整个过程中上调，29个下调；10个在不同时点分别表现上调或下调。这些基因涉及到生长素的运输、响应、诱导、合成和降解等多个生物学过程。对BG906698、BQ281752、CD454626、CD864552、CD938626、BJ233383和AY543630基因的半定量RT-PCR验证结果表明，它们的表达变化与基因芯片的结果基本一致。质膜H+-ATPase基因(AY543630)在2 h时间点即有较高强度表达，然后下降，24 h降至最低水平，表明该基因在小麦成熟胚脱分化的启动中可能有重要作用。

关键词: 生长素相关基因, 小麦成熟胚脱分化, 质膜H+-ATPase基因, 基因芯片

Abstract:

The quality of dedifferentiation determines the capacity of callus regeneration. Mature embryo is considered to be of the most potential for genetic transformation, but its transformation efficiency is still lower now.To elucidate the mechanism of mature wheat embryo dedifferentiation, we studied the expression profile of genes in dedifferentiation using Affymetrix microarray technique in mature embryos of wheat cultivar Yumai 18 cultured on MS medium supplemented with 2 mg L-1 of 2,4-dichlorophenolxyacetic acid (2,4-D) at different time points of 2, 6, 12, 24, and 72 h. Using online tools at NCBI, DATF, and DRTF websites, 80 auxin-related genes changed at least at one time point were detected with the 41 up-regulated and 29 down-regulated during the whole period. The remained 10 genes showed up-regulation or down-regulation at different time points. Up-regulated genes were more than down-regulated genes, especially at 24 h and 72 h points. These genes were involved in several biological processes, such as transportation, response, induction, synthesis, and degeneration of auxin. Using semi-quantative RT-PCR technique, the expression changes of seven genes, coded as BG906698, BQ281752, CD454626, CD864552, CD938626, BJ233383, and AY543630, were confirmed to be consistent with the result by microarray. Gene of plasma membrane (PM) H+-ATPase (GenBank accession number: AY543630) expressed at a high level at the 2 h time point, and then decreased gradually till the minimum at the 24 h time point. This result implied that PM H+-ATPase gene might be a trigger in dedifferentiation of mature embryo in wheat.

Key words: Auxin-related gene, Dedifferentiation of mature embryo, wheat, Plasma membrane H+-ATPase gene, Microarray

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