摘要
本研究以野生蕉胚性悬浮系为材料,将胚性细胞增殖培养基上的香蕉胚性悬浮系作为对照,体胚诱导培养基上的培养材料为处理,利用抑制性消减杂交(suppression subtractive hybridization,SSH)技术,构建野生蕉胚性悬浮系激素诱导的胚胎发育差异表达cDNA文库,随机挑选阳性克隆并进行生物信息学分析。结果显示,聚类得到62条非冗余EST。经Blast比对后发现其中18条没有对应功能的相似序列,46条在GenBank中有同源序列,其中12条为功能未知的基因。在文库中功能已知的ESTs中,这些基因涉及基础代谢(primary metabolism)、蛋白结合(protein binding)、转录(transport)细胞分化(cell differentiation)等功能。文库中发现了一些可能与体细胞胚胎发育相关的基因如TPD1基因、热激蛋白基因、谷胱甘肽-S-转移酶基因、锌指蛋白基因以及细胞壁富含羟脯氨酸糖蛋白基因等。试验结果为获得香蕉体细胞胚胎发育相关基因全长并验证这些基因的功能提供依据。
A subtracted cDNA library of was constructed with the embryogenic cell suspensions （ECS） of the wild banana by using Suppression Subtractive Hybridzation （SSH）. The ECS subcultured in cell multiplication medium were harvested as control materials, and the ECS subcultured in embryo development medium were harvested as the treated materials. Positive clones were randomly selected and sequenced from the SSH-cDNA library. After the removal of duplication and sequencing of low-quality sequences, 62 high quality sequences were obtained, According to BLAST screening and functional annotation, the results revealed that 46 ESTs have high homology with known genes in GenBank, 12 ESTs homologous sequence were not found, presumably may be new genes. The differentially expressed genes are involved in primary metabolism, protein binding, transport, cell differentiation and et al. Among these ESTs, TPD1, heat shock proteins, and zinc finger protein and cell wall hydroxyproline-rich glycoprotein are probably related to embryo development of banana. The SSH cDNA library can be further used for gene cloning and identification for the development-related genes in banana.
出处
《分子植物育种》
CAS
CSCD
2011年第5期592-598,共7页
Molecular Plant Breeding
基金
国家自然科学基金(31000903)
广东省自然科学基金(10451064001006326)
广东省农业科学院院长基金(20090104)共同资助
