【摘要】：芒果炭疽病(Colletotrichum gloeosporioides)是為害芒果的重要病害之一,為了明確小柱孢酮脫水酶基因SCD1與芒果炭疽病菌致病力之間的相互關(guān)系,本研究以芒果炭疽病菌DNA為模板,利用同源克隆技術(shù)擴(kuò)增SCD1,分析其序列特征、推測(cè)了其蛋白的保守結(jié)構(gòu)域,并借助In-Fusion~汶 HD Cloning Kit技術(shù)進(jìn)行敲除載體構(gòu)建。結(jié)果表明,該基因DNA和cDNA全長(zhǎng)分別為796 bp、564 bp,編碼區(qū)有兩個(gè)內(nèi)含子(大小為52 bp,180 bp),推測(cè)編碼187個(gè)氨基酸,其分子量約為21.52 kD,等電點(diǎn)PI為5.90,與NCBI網(wǎng)站中已公布的基因進(jìn)行Blastp比對(duì),發(fā)現(xiàn)該序列與香蕉炭疽菌(C.musae)、無(wú)花果炭疽菌(C.caricae)(登錄號(hào):GQ266389.1,GQ266386.1)的小柱孢酮脫水酶基因相似性分別為98%和97%。該基因的敲除載體pSCDGH-1已構(gòu)建成功,為下一步獲得SCD1基因敲除突變體,研究該基因功能打下了材料基礎(chǔ)。
[Abstract]:Mango anthracnose (Colletotrichum gloeosporioides) is one of the important diseases of mango. In order to clarify the relationship between SCD1 gene and pathogenicity of mango anthracnose, DNA of mango anthracnose was used as a template in this study. Homologous cloning technique was used to amplify SCD1, to analyze its sequence characteristics. The conserved domain of its protein was speculated, and the knockout vector was constructed by means of In-Fusion~ HD Cloning Kit technique. The results showed that the full length of DNA and cDNA were 796 bp,564 bp, respectively. There were two introns (52 bp,180 bp), in size) encoding 187 amino acids, the molecular weight of which was about 21.52 kD, isoelectric point PI was 5.90. Blastp alignment with genes published on the NCBI website revealed that the sequence was linked to banana anthrax (C.musae) and fig anthrax (C.caricae) (accession number: GQ266389.1,) The similarity of cyclosporine dehydratase gene in GQ266386.1 was 98% and 97%, respectively. The knockout vector pSCDGH-1 has been successfully constructed, which lays the material foundation for the next step to obtain the mutant of SCD1 gene knockout and to study the function of the gene.
【作者單位】： 海南大學(xué)環(huán)境與植物保護(hù)學(xué)院海南大學(xué)熱帶作物種質(zhì)資源保護(hù)與開(kāi)發(fā)利用教育部重點(diǎn)實(shí)驗(yàn)室;中國(guó)熱帶農(nóng)業(yè)科學(xué)院環(huán)境與植物保護(hù)研究所農(nóng)業(yè)部熱帶作物有害生物綜合治理重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金(31460455) 公益性行業(yè)(農(nóng)業(yè))科研專項(xiàng)項(xiàng)目 芒果產(chǎn)業(yè)技術(shù)研究與示范(201203092-2)共同資助
【分類號(hào)】：S436.67
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本文編號(hào)：2411295