利用cDNA-AFLP技术分析小麦成株抗条锈性差异基因表达特征

doi:10.3724/SP.J.1006.2010.00401

Abstract

Abstract:

Wheat (Triticum aestivum L.) stripe rust is one of the most devastating diseases of wheat throughout the world. Adult plant resistance (APR) to stripe rust in wheat conferring durable resistance, thus, plays a pivotal role in the control of the disease. In the present study, to elucidate molecular mechanism of wheat APR to stripe rust, we conducted extensive transcription profiling of adult-plant wheat cultivar Xingzi 9104 infected by Puccinia striiformis Westend f. sp. tritici Erikss. pathotype CY32 using cDNA-AFLP technique. We analyzed transcription profiling of the incompatible reaction across nine sampling time points within five days after inoculation. Of the total 32,320 transcript derived fragments (TDFs) obtained using cDNA-AFLP with 64 primer pairs, 2201 (6.81%) displayed altered expression patterns after inoculation, of which 926 showed up-regulated and 1275 down-regulated. Three hundred and thirty differentially expressed TDFs produced reliable sequences after cloning and sequencing, of which 259 expressed sequence tags (ESTs) of unigenes were obtained after assembling, designated from aTaPST1 to aTaPST259, deposited in GenBank with accessions numbers from FL645754 to FL646011 and FL646262. BLASTX analyses and functional annotations were then performed and the results revealed that the 95 ESTs had predicted gene products mainly implicated in energy (11.20%), metabolism (4.63%), transcription (3.86%), disease/defense (3.86%), protein destination and storage (3.09%), protein synthesis and cell growth (each accounted for 2.32%), and signal transduction (1.54% of the sequenced total 259 ESTs). Six differential genes related to disease/defense, transcription, and signal transduction were chosen for further qRT-PCR expression patterns, which confirmed the cDNA-AFLP profiles. Our results indicated that wheat APR to stripe rust involved in multifaceted biochemical and physiological reactions, including concerted regulation of the genes involved in different pathways like disease/defense, transcription, protein metabolism, signal transduction, as well as abiotic stresses. These results provide information for further elucidation of molecular mechanism of wheat APR to stripe rust.

Key words: Wheat, Stripe rust fungus, Adult-plant resistance, Gene expression, cDNA-AFLP, qRT-PCR

ZHANG Gang,DONG Yan-Ling,XIA Ning,ZHANG Yi,WANG Xiao-Jie,QU Zhi-Peng,LI Yi-Min,.

cDNA-AFLP Analysis Reveals Differential Gene Expression in Wheat Adult-Plant Resistance to Stripe Rust

[J].Acta Agron Sin, 2010, 36(3): 401-409.

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