小麦抗白粉病SSH-cDNA文库中差异基因的表达模式

doi:10.3724/SP.J.1006.2008.02121

Abstract

Abstract:

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important fungal diseases of common wheat (Triticum aestivum L.) worldwide and causes severe yield losses. Wheat germplasm N9436, developed by our research group, is a resistant material to powdery mildew. In the present study, a suppression subtraction hybridization (SSH) cDNA library was constructed with cDNA from N9436 leaf inoculated by Blumeria graminis as the tester and cDNA from N9436 healthy leaf as the driver. A total of 140 positive clones were randomly chosen from the SSH-cDNA library and were amplified with sp6 and t7 primers to examine the insert size, which ranged from 200 to 1 000 bp with an average of 238 bp. After screening repeat and redundancy sequences, 94 expressed sequence tags (ESTs) were acquired. Protein homology search in nr-protein database revealed that 49 ESTs were highly homologous with known proteins functioning in primary metabolism (2%), energy metabolism (24%), cell structure (2%), transcription (2%), protein synthesis and processing (16%), transport (4%), signal transduction (4%), and disease resistance and defenses (30%). Compared with the EST sequences among the SSH-cDNA library, 2 ESTs (GenBank accession number: EX567369 and EX567360) were highly similar to pathogenesis-related protein (Z25-1) and glutathione-S-transferase (Z440-1), respectively. On the basis of the EST sequences, 2 pairs of primers were designed and used to detect the expression differences of the 2 genes when inoculating with powdery mildew by Reverse Transcriptase PCR (RT-PCR). Z25-1 and Z440-1 were both expressed under inoculation. Their expression amounts were the largest in 72 h after inoculation with powdery mildew. But their expression trends were different. At early stage of inoculation (24 h after inoculation), the expression amount of Z440-1 was very a little, then increased in 72 h, and finally decreased in 96 h. The expression amount of Z25-1 increased from 24 h to 72 h after inoculation and decreased after 96 h. It suggests that pathogenesis-related protein and glutathione-S-transferase belong to inducible expression genes and are involved in the resistance to powdery mildew.

Key words: Wheat, Powdery mildew, Suppression subtraction hybridization (SSH), Reverse transcriptase PCR

WU Jin-Hua;HU Yin-Gang;WANG Xin-Ru;ZHANG Hong;WANG Chang-You;WANG Qiu-Ying;JI Wan-Quan. Expression Pattern of Special Genes Resistant to Powdery Mildew (Blumeria graminis f. sp. tritici) in SSH-cDNA Library of Wheat[J].Acta Agron Sin, 2008, 34(12): 2121-2125.

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Expression Pattern of Special Genes Resistant to Powdery Mildew (Blumeria graminis f. sp. tritici) in SSH-cDNA Library of Wheat

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