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以“巴西”蕉(Musa acuminata L.AAA group cv.Brazilian)为试材,利用同源克隆法获得4个颗粒结合淀粉合成酶I(Granule-Bound Starch Synthase I,GBSSI)基因家族成员的cDNA全长,运用MEGA 5.05软件进行聚类分析,并通过quantitative real-time PCR(qPCR)技术检测MaGBSSI基因家族成员在香蕉果实不同发育时期及成熟阶段和不同组织中的表达情况。结果表明:香蕉4个MaGBSSI基因家族成员MaGBSSI-1、MaGBSSI-2、MaGBSSI-3、MaGBSSI-4的cDNA全长分别为1 851、1 851、675、1 845bp,登录号分别为KF512020、KF512021、KF512022、KF512023。聚类分析发现,香蕉MaGBSSI基因家族与其它植物GBSSI基因同源性达65%。qPCR分析发现,MaGBSSI-1、MaGBSSI-2、MaGBSSI-4在香蕉根、球茎、叶片和苞片等营养器官中明显上调表达,而MaGBSSI-3在花、果皮、果肉等生殖器官中表达量比较高,在根、球茎及叶片中几乎不表达。香蕉果实发育过程中,MaGBSSI-1、MaGBSSI-2、MaGBSSI-4在0~30d明显上调表达,而MaGBSSI-3在香蕉果实发育30~60d表达量比较高,在0~30d几乎不表达,且随着果实的成熟,MaGBSSI-3表达量逐渐下降。研究结果为阐明香蕉果实的直链淀粉生物合成及降解机制并对其进行表达调控奠定了基础。
Using the method of homologous cloning, four members of the Granule-Bound Starch Synthase I (GBSSI) gene family were obtained from Musa acuminata L.AAA group cv.Brazilian The full length of cDNA was analyzed by using MEGA 5.05 software. The quantitative real-time PCR (qPCR) was used to detect the expression of MaGBSSI genes in different development stages, mature stages and different tissues of banana fruit. The results showed that the full-length cDNAs of MaGBSSI-1, MaGBSSI-2, MaGBSSI-3 and MaGBSSI-4 were 1 851, 1 851, 675 and 1 845 bp respectively. The accession numbers were KF512020, KF512021, KF512022, KF512023. Cluster analysis showed that the banana MaGBSSI gene family shared 65% homology with other plant GBSSI genes. qPCR analysis showed that MaGBSSI-1, MaGBSSI-2 and MaGBSSI-4 were significantly up-regulated in vegetative organs such as banana root, bulb, leaf and bracts, while MaGBSSI-3 expression in reproductive organs such as flower, peel and pulp High, barely expressed in roots, bulbs and leaves. MaGBSSI-1, MaGBSSI-2 and MaGBSSI-4 were significantly up-regulated during 0-30 d in banana fruit development, whereas MaGBSSI-3 expression was relatively high at 30-60 d in banana fruit development and hardly expressed in 0-30 d With the maturation of fruit, the expression of MaGBSSI-3 gradually decreased. The results laid the foundation for elucidating the mechanism of amylose biosynthesis and degradation in banana fruit and regulating its expression.