【摘要】：C2H2型鋅指蛋白在植物生長發(fā)育、非生物脅迫響應(yīng)過程中起關(guān)鍵作用。基于實驗室前期工作,分析鹽脅迫下陸地棉根系抑制性差減雜交文庫,篩選得到1個鹽脅迫應(yīng)答基因GhC2H2。結(jié)合RACE(Rapid amplification of cDNA ends)和RT-PCR(Reverse transcription-polymerase chain reaction)技術(shù)克隆該基因全長,通過瞬時表達(dá)分析其細(xì)胞定位,采用Real time-PCR分析其在鹽脅迫下的陸地棉根、莖、葉中的表達(dá)模式,并測定了轉(zhuǎn)入該基因的株系在正常土壤和鹽堿地的鈴重、衣分和纖維品質(zhì)指標(biāo)。該基因GenBank登錄號為HM002632,編碼區(qū)全長819 bp,編碼272 aa;其編碼蛋白包含2個C2H2鋅指結(jié)構(gòu)域,定位于細(xì)胞核;該基因在不同組織中均受鹽脅迫誘導(dǎo)上調(diào)表達(dá),且在脅迫誘導(dǎo)初期上調(diào)量最大,推測該基因參與早期鹽脅迫應(yīng)答。對轉(zhuǎn)基因后代株系性狀分析表明,該基因的過表達(dá)可以提高轉(zhuǎn)基因棉花衣分、改良纖維品質(zhì),推測GhC2H2既參與鹽脅迫應(yīng)答過程,也參與纖維發(fā)育調(diào)控。
[Abstract]:C2H2 zinc finger protein plays a key role in plant growth and development, abiotic stress response. Based on the previous work in laboratory, the subtractive hybridization library of root inhibition of Upland cotton under salt stress was analyzed, and a salt stress response gene GhC2H2. was screened out. The full length of the gene was cloned by RACE (Rapid amplification of cDNA ends) and RT-PCR (Reverse transcription-polymerase chain reaction), and its cellular localization was analyzed by transient expression analysis. The expression patterns of the gene in root, stem and leaf of upland cotton under salt stress were analyzed by Real time-PCR. The boll weight, lint percentage and fiber quality index of the transgenic lines in normal soil and saline soil were determined. The GenBank accession number of the gene was 819 bp, encoding 272 aa;. The encoded protein contained two C2H2 zinc-finger domains and was located in the nucleus. The expression of the gene was up-regulated in different tissues induced by salt stress, and the up-regulation was the highest in the early stage of stress induction. It was speculated that the gene was involved in the early salt stress response. The analysis of the characters of transgenic progenies showed that the overexpression of the gene could improve lint score and fiber quality of transgenic cotton. It was inferred that GhC2H2 was involved in both salt stress response and fiber development regulation.
【作者單位】： 中國農(nóng)業(yè)大學(xué)農(nóng)學(xué)院/雜種優(yōu)勢研究與利用教育部重點實驗室/作物遺傳改良北京市重點實驗室;河北省農(nóng)林科學(xué)院棉花研究所/農(nóng)業(yè)部黃淮海半干旱區(qū)棉花生物學(xué)與遺傳育種重點實驗室;湖北省農(nóng)業(yè)科學(xué)院經(jīng)濟(jì)作物研究所;
【基金】：國家高技術(shù)研究發(fā)展計劃(863計劃)重大項目(2006AA10A108) 國家科技重大專項——轉(zhuǎn)基因生物新品種培育(2014ZX08005-004B)
【分類號】：S562
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本文編號：2393206