【摘要】：WRKY家族轉(zhuǎn)錄因子參與植物各種生物脅迫和非生物脅迫反應(yīng),同時也參與形態(tài)建成、物質(zhì)代謝、種子休眠、衰老等過程。本研究在前期紅掌疫病誘導(dǎo)轉(zhuǎn)錄組數(shù)據(jù)中篩選到一個WRKY家族基因,RT-PCR擴增到1 658 bp全長cDNA,包含一個1 035 bp的開放閱讀框,生物信息學(xué)分析預(yù)測此基因是一個典型的WRKY轉(zhuǎn)錄因子,命名為Aan WRKY1(NCBI No.KY597634)。qRT-PCR證實該基因葉片中受疫病侵染上調(diào)表達(dá),同時能夠被SA、JA、ABA、ETH、GA等激素信號分子誘導(dǎo),非處理條件下在花梗和莖中表達(dá)量最高。通過農(nóng)桿菌介導(dǎo)洋蔥表皮侵染法,瞬時表達(dá)AanWRKY1-eGFP,顯示該基因編碼蛋白定位于細(xì)胞核,進(jìn)一步酵母單雜交實驗,證明該基因具有離體轉(zhuǎn)錄激活活性。研究結(jié)果表明紅掌AanWRKY1基因能夠響應(yīng)細(xì)菌性疫病及其他脅迫反應(yīng),具有應(yīng)激轉(zhuǎn)錄調(diào)控的作用,該基因的鑒定為進(jìn)一步解析其分子病理學(xué)作用機理提供依據(jù)。
[Abstract]:WRKY family transcription factors are involved in plant stress and abiotic stress responses, but also participate in morphogenesis, substance metabolism, seed dormancy, senescence and other processes. In this study, an WRKY family gene was amplified by RT-PCR from the primordial transcripts of blight. The full-length cDNA, of 1 658 bp contained an open reading frame of 1 035 bp. Bioinformatics analysis predicted that this gene was a typical WRKY transcription factor. Aan WRKY1 (NCBI No.KY597634). QRT-PCR confirmed that the gene was up-regulated by blight infection in leaves and could be induced by hormone signaling molecules such as SA,JA,ABA,ETH,GA, and the highest expression was found in pedicels and stems under untreated conditions. Through Agrobacterium tumefaciens mediated onion epidermal infection, the transient expression of AanWRKY1-eGFP, showed that the gene encoded protein was located in the nucleus. Further yeast single hybrid experiments showed that the gene had in vitro transcriptional activation activity. The results showed that the AanWRKY1 gene could respond to bacterial blight and other stress responses and play a role in stress transcriptional regulation. The identification of the gene provides a basis for further elucidation of its molecular pathological mechanism.
【作者單位】： 海南大學(xué)農(nóng)學(xué)院;中國熱帶農(nóng)業(yè)科學(xué)院熱帶作物品種資源研究所;
【基金】：國家自然科學(xué)基金青年項目(31201652);國家自然科學(xué)基金面上項目(31471902)共同資助
【分類號】：S436.8
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本文編號：2253005