發(fā)布時(shí)間：2018-06-29 18:14

  本文選題：陸地棉 + b��； 參考：《中國農(nóng)業(yè)科技導(dǎo)報(bào)》2016年05期

【摘要】：通過電子克隆技術(shù)從陸地棉中分離了一個(gè)Gh MYC4基因(Gen Bank登錄號:KF751654),該基因編碼的蛋白屬于b HLH類轉(zhuǎn)錄因子。生物信息學(xué)分析結(jié)果顯示:該基因完整開放閱讀框有1 650 bp,編碼549個(gè)氨基酸。Gh MYC4蛋白的N-端具有MYC蛋白家族特有的b HLH-MYC-N保守結(jié)構(gòu)域,C端具有高度保守的DNA結(jié)合域即b HLH結(jié)構(gòu)域。同源比對和系統(tǒng)進(jìn)化樹分析結(jié)果顯示Gh MYC4蛋白與陸地棉b HLH轉(zhuǎn)錄因子和可可b HLH轉(zhuǎn)錄因子的親緣關(guān)系最近,與其他物種的MYC2家族成員具有很大的相似性。通過Real time-PCR檢測該基因的組織表達(dá)特異性,結(jié)果顯示該基因在陸地棉的根部和葉片中高效表達(dá),纖維中微量表達(dá)。在擬南芥原生質(zhì)體中進(jìn)行該基因的瞬時(shí)表達(dá),證實(shí)了Gh MYC4蛋白定位于細(xì)胞核。轉(zhuǎn)Gh MYC4的擬南芥受到高鹽和干旱脅迫后相比于對照在表型上表現(xiàn)出很好的抗旱耐鹽性能,其葉片相對含水量、可溶性糖含量及脯氨酸含量顯著高于對照,MDA含量顯著低于對照。該結(jié)果為培育抗旱耐鹽棉花新品種提供了新的基因來源。
[Abstract]:A Gh MYC4 gene (GenBank accession number: KF751654) was isolated from upland cotton by electronic cloning technique. The protein encoded by this gene belongs to bHLH transcription factor. Bioinformatics analysis showed that the complete open reading frame of this gene was 1 650 BP, encoding 549 amino acids. Gh MYC4 protein had a highly conserved DNA binding domain, b HLH domain, in the C-terminal of b HLH-MYC-N conserved domain (b HLH-MYC-N), which is unique to the MYC protein family. The results of homology comparison and phylogenetic tree analysis showed that Gh MYC4 protein was closely related to b HLH transcription factors and cocoa b HLH transcription factors in upland cotton, and was similar to the MYC2 family members of other species. The tissue expression specificity of the gene was detected by Real time-PCR. The results showed that the gene was highly expressed in the roots and leaves of upland cotton, and the micro-expression was found in the fiber. Transient expression of the gene in Arabidopsis protoplasts confirmed that Gh MYC4 protein was located in the nucleus of Arabidopsis thaliana. Arabidopsis thaliana transgenic with Gh MYC4 showed better drought resistance and salt tolerance than the control after high salt stress and drought stress. The relative water content, soluble sugar content and proline content in leaves of Arabidopsis thaliana transgenic with Gh MYC4 were significantly higher than those in control group. The results provide a new gene source for breeding new drought-resistant and salt-tolerant cotton varieties.
【作者單位】： 中國農(nóng)業(yè)科學(xué)院生物技術(shù)研究所;西北大學(xué)生命科學(xué)學(xué)院;深圳市鐵漢生態(tài)環(huán)境股份有限公司;
【基金】：國家轉(zhuǎn)基因生物新品種培育重大專項(xiàng)(2014ZX08005-004) 廣東省省級科技計(jì)劃項(xiàng)目(2015B090904008)資助
【分類號】：S562;Q943.2

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