【摘要】：該研究以擬南芥抗逆基因At1g67520為探針,利用海島棉ESTs數(shù)據(jù)庫(kù),通過(guò)電子克隆獲得海島棉RLCK家族基因GbRLCK10,解析該基因組結(jié)構(gòu),并結(jié)合qRT-PCR技術(shù)分析該基因mRNA的組織表達(dá)特征以及在不同脅迫誘導(dǎo)下的表達(dá)模式,為揭示RLCK家族基因在海島棉中的表達(dá)調(diào)控及作用機(jī)制提供理論依據(jù)。結(jié)果顯示:(1)獲得海島棉類(lèi)受體胞質(zhì)激酶(RLCK)基因,其開(kāi)放閱讀框(ORF)為1 179bp,編碼392個(gè)氨基酸,具有典型的Serine/Threonine結(jié)構(gòu)域,屬于RLCK家族,與GaRLCK10(XP_017604046.1)親緣關(guān)系較近,命名為GbRLCK10(登錄號(hào)2022184),且該基因由5個(gè)外顯子和4個(gè)內(nèi)含子組成。(2)實(shí)時(shí)熒光定量(qRT-PCR)檢測(cè)顯示,GbRLCK10基因在抗病品種‘新海21’和感病品種‘新海14’的根、莖、葉中均有表達(dá);當(dāng)黃萎病菌誘導(dǎo)后,GbRLCK10基因在抗病品種中對(duì)于病原菌的響應(yīng)時(shí)間早于感病品種,且對(duì)黃萎病菌響應(yīng)更強(qiáng)烈,推測(cè)該基因參與棉花對(duì)黃萎病的響應(yīng);鹽(NaCl)、干旱(PEG-6000)處理‘新海21’后,GbRLCK10基因在NaCl處理下響應(yīng)時(shí)間要早于PEG-6000處理,但對(duì)PEG-6000處理響應(yīng)更強(qiáng)烈;分別用4種激素處理‘新海21’后,GbRLCK10均能被誘導(dǎo)表達(dá),且在水楊酸(SA)處理后表現(xiàn)為先增加后下降再增加趨勢(shì),在乙烯(ET)處理后表達(dá)量為持續(xù)上升趨勢(shì),在茉莉酸甲酯(MeJA)處理后呈先升高然后下降的趨勢(shì),但GbRLCK10基因?qū)Τ嗝顾?GA3)響應(yīng)不明顯。研究表明,GbRLCK10基因具有RLCK基因家族典型特征,該基因隨黃萎病菌、NaCl、干旱、激素處理時(shí)間推移而發(fā)生變化,推測(cè)GbRLCK10基因可能參與了棉花對(duì)黃萎病菌、NaCl、干旱、激素脅迫的應(yīng)答反應(yīng),但其功能仍需進(jìn)一步研究。
[Abstract]:Using Arabidopsis thaliana (Arabidopsis thaliana) stress resistance gene At1g67520 as probe and island cotton ESTs database, the genomic structure of island cotton RLCK family gene GbRLCK10, was obtained by electronic cloning. The expression characteristics of mRNA and the expression patterns under different stress were analyzed by qRT-PCR technique, which provided a theoretical basis for revealing the regulation and mechanism of the expression of RLCK family genes in island cotton. The results showed that: (1) the receptor cytoplasmic kinase (RLCK) gene was obtained. Its open reading frame (ORF) was 1179 BP, encoding 392 amino acids. It had a typical Serine/Threonine domain and belonged to the RLCK family. The gene was closely related to GaRLCK10 (XP_017604046.1) and was named GbRLCK10 (accession number 2022184), and the gene was composed of 5 exons and 4 introns. (2) Real-time fluorescence quantitative (qRT-PCR) analysis showed that the gene was highly related to GbRLCK10 (accession number 2022184), and the gene was composed of 5 exons and 4 introns. GbRLCK10 gene was expressed in the roots, stems and leaves of the resistant variety 'Xinhai 21' and the susceptible variety 'Xinhai 14'. When Verticillium wilt was induced, the response time of GbRLCK10 gene to pathogen in resistant varieties was earlier than that of susceptible variety, and the response to Verticillium wilt was stronger. It was speculated that the GbRLCK10 gene was involved in the response of cotton to Verticillium wilt. The response time of GbRLCK10 gene under NaCl treatment was earlier than that of PEG-6000 treatment after salt (NaCl), drought (PEG-6000) treatment, but the response to PEG-6000 treatment was stronger. After treated with four hormones, the expression of GbRLCK10 was induced, and the expression of GbRLCK10 increased first, then decreased and then increased after treatment with salicylic acid (SA), and the expression level increased continuously after treatment with ethylene (ET). After (MeJA) treatment with methyl jasmonate, the GbRLCK10 gene increased first and then decreased, but the response of GbRLCK10 gene to gibberellin (GA3) was not obvious. The results showed that GbRLCK10 gene had the typical characteristics of RLCK gene family, and it changed with Verticillium wilt, NaCl, drought and hormone treatment. It was suggested that GbRLCK10 gene might be involved in cotton drought to Verticillium wilt and NaCl,. Response to hormone stress, but its function still needs further study.
【作者單位】： 新疆農(nóng)墾科學(xué)院生物技術(shù)研究所/作物種質(zhì)創(chuàng)新與基因資源利用兵團(tuán)重點(diǎn)實(shí)驗(yàn)室;中國(guó)農(nóng)業(yè)科學(xué)院生物技術(shù)研究所;
【基金】：兵團(tuán)應(yīng)用基礎(chǔ)研究計(jì)劃(2016AG005) 國(guó)家重點(diǎn)研發(fā)計(jì)劃(2016YFD0100203-6)
【分類(lèi)號(hào)】：Q943.2;S562
，

本文編號(hào)：2375925