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作物学报 ›› 2009, Vol. 35 ›› Issue (7): 1188-1193.doi: 10.3724/SP.J.1006.2009.01188

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

应用基因芯片分析红蚰麦白粉菌胁迫条件下的基因表达谱

王俊美1,2,刘红彦1,2,*,徐红明1,王飞1,高素霞1,康振生2,3,*   

  1. 1河南省农业科学院植物保护研究所,河南郑州450002;2西北农林科技大学植保资源与病虫害治理教育部重点开放实验室,陕西杨凌712100;3西北农林科技大学山西省农业分子生物学重点实验室，陕西杨凌712100
  • 收稿日期:2009-01-07 修回日期:2009-03-10 出版日期:2009-07-12 网络出版日期:2009-05-18
  • 通讯作者: 刘红彦,E-mail:liuhy1219@163.com,Tel:0371-65730166;康振生,E-mail:kangzs@nwsuaf.edu.cn,Tel:029-87091312
  • 基金资助:

    本研究由河南省杰出青年基金（04120001400），西北农林科技大学植保资源与病虫害治理教育部重点开放实验室资助项目，国家“十一五”支撑计划项目（2006BAD08A05)资助。

Expression Profile of Landrace Hongyoumai Infected by Blumeria graminis f.sp.tritici Using Gene Microarray

WANG Jun-Mei1,2,LIU Hong-Yan1,2,*,XU Hong-Ming1,WANG Fei1,GAO Su-Xia1,KANG Zhen-Sheng23*   

  1. 1Institute of Plant Protection，Henan Academy of Agricultural Sciences，Zhengxhou 450002，China；2Key Laboratory of Plant Protection Resources and Pest Management，Ministry of Education，Northwest A&F University，Yangling 712100，China；3Shaanxi Key Laboratory of Molecular Biology for Agriculture，Northwest A&F University，Yangling 712100，China
  • Received:2009-01-07 Revised:2009-03-10 Published:2009-07-12 Published online:2009-05-18
  • Contact: LIU Hong-yan,E-mail:liuhy1219@163.com,Tel: 0371-65730166;KANG Zhen-Sheng,E-mail:kangzs@nwsuaf.edu.cn,Tel:029-87091312

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2255

被引次数

13

摘要：

小麦农家品种红蚰麦携带1对显性抗白粉病新基因，暂命名为Pmhym。为了深入了解该品种抗病的分子机制，用豫麦13/红蚰麦的F3代纯合抗病株系构建抗池，并对白粉菌接种后24 h的基因表达谱，采用Affymetrix小麦基因芯片进行分析，以白粉菌接种0 h为对照。在61 127基因微矩阵点中，有效差异表达Ratio值≥2或≤0.5的基因共5 282个，其中上调基因2 553个，下调2 729个。对上调序列的分类表明，功能明确的序列中39.81%与抗病/防御相关；下调表达基因中以能量代谢和抗病/防御相关基因比例最高，分别占26.71%和19.65%。上调表达在8倍以上的序列中81个的功能已知，其中包括病程相关基因、防卫反应基因、生成或清除活性氧的基因，以及抗病信号转导基因等。选取上调和下调表达8倍以上的共13个探针序列进行实时定量PCR验证，表明基因芯片数据具有良好的重复性。

关键词: 小麦, 抗白粉病基因, 基因芯片, 表达序列标签, 实时定量PCR

Abstract:

Wheat landrace Hongyoumai carries one dominant resistance gene for powdery mildew (Blumeria graminis), tentatively designated Pmhym. To further elucidate molecular mechanism of the resistance to powdery mildew in Hongyoumai, a resistant pool was constructed with four homogenous resistant lines from the F3 generation of Yumai 13/Hongyoumai. The wheat seedlings were inoculated with single spore isolates of B. graminis f. sp. tritici, and at 0 and 24 h after inoculation the gene expression profile was analyzed using Affymetrix wheat microarray. There were approximately 5 282 expressed genes (with Log ratio ≥ 2 or ≤ 0.5) among total 61 127 genes set in the microarray plate, including 2 553 up-regulated and 2729 down-regulated genes or expressed sequence tags (ESTs). In the up-regulated ESTs whose functions have been known, 39.81% were disease/defense genes. In the down-regulated ESTs, energy genes and disease/defense genes accounted for 26.71% and 19.65%, respectively. A total of 81 ESTs were up-regulated more than eight times, in which most were related to disease resistance, such as pathogenesis-related protein, defense genes, genes producing or eliminating reactive oxygen species, and genes involved in signal transduction. Quantitative RT-PCR (qRT-PCR) was carried out with 13 probe sets up-regulated and down-regulated more than eight times which validated a good reproducibility of microarray analysis.

Key words: Wheat, Powdery mildew resistance gene, Gene chip, Expressed sequence Tag（EST）, Real-time quantitative RT-PCR(q-PCR)

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