龙眼UGD6基因克隆及其表达特性分析
发布时间：2018-11-10 23:50
【摘要】：该试验采用RT-PCR和RACE技术,对龙眼多糖合成的关键基因尿苷二磷酸-葡萄糖6-脱氢酶基因(DlUGD6)进行分离克隆、生物信息学分析和亚细胞定位研究,并采用qRT-PCR技术,对其在龙眼体细胞胚胎发生、合子胚发育及不同组织器官中的表达模式进行分析。结果表明:(1)DlUGD6基因的cDNA序列全长1 860bp,包含开放阅读框1 443bp,编码480个氨基酸(GenBank登录号KU198438);生物信息学分析显示,DlUGD6属于稳定的酸性亲水蛋白,不含信号肽,具有跨膜结构和3个典型的保守结构域,属于UDP-葡萄糖/GDP-甘露糖脱氢酶家族;进化树分析表明,DlUGD6与柑橘亲缘关系较近。(2)洋葱内表皮GFP荧光定位观察发现,DlUGD6定位于细胞质;qRT-PCR结果显示,DlUGD6在龙眼非胚性愈伤组织中表达量相对较高,且在其他体胚发育阶段也均有稳定表达;在合子胚发育中子叶胚形成后第8天(S3)和第24天(S7)时表达量最高,整体呈"W"型;在不同组织器官中,DlUGD6在花药和茎中的表达量最高,且整体上生殖器官中的表达水平高于营养器官。研究认为,DlUGD6基因可能参与龙眼生长发育各个阶段中细胞壁多糖合成。
[Abstract]:The key gene of longan polysaccharide synthesis was isolated and cloned by RT-PCR and RACE, bioinformatics analysis and subcellular localization were carried out, and qRT-PCR technique was used. Its expression patterns in somatic embryogenesis, zygotic embryo development and different tissues and organs of longan were analyzed. The results showed that: (1) the cDNA sequence of DlUGD6 gene was 1 860 BP, including open reading frame 1443 BP, encoding 480 amino acids (GenBank accession number KU198438); Bioinformatics analysis showed that DlUGD6 was a stable acidic hydrophilic protein with no signal peptide, and had transmembrane structure and three typical conserved domains, belonging to the UDP- glucose / GDP- mannose dehydrogenase family. Phylogenetic tree analysis showed that DlUGD6 was closely related to citrus. (2) GFP fluorescence localization of onion inner epidermis showed that DlUGD6 was located in cytoplasm; QRT-PCR results showed that the expression of DlUGD6 was relatively high in non-embryogenic callus of longan and stable in other somatic embryogenesis. At the 8th (S3) and 24th (S7) days after the development of zygotic embryos, the highest expression level was observed, and the whole expression was "W" type. The expression of DlUGD6 in anthers and stems was the highest in different tissues and organs, and the overall expression level of DlUGD6 in reproductive organs was higher than that in vegetative organs. It is suggested that DlUGD6 gene may be involved in the synthesis of polysaccharides from the cell wall of longan in various stages of growth and development.
【作者单位】： 福建农林大学园艺植物生物工程研究所;
【基金】：国家自然科学基金(31572088,31272149) 福建省重大科技专项(2015NZ-0002-1)
【分类号】：Q943.2;S667.2
，
本文编号：2323924
[Abstract]:The key gene of longan polysaccharide synthesis was isolated and cloned by RT-PCR and RACE, bioinformatics analysis and subcellular localization were carried out, and qRT-PCR technique was used. Its expression patterns in somatic embryogenesis, zygotic embryo development and different tissues and organs of longan were analyzed. The results showed that: (1) the cDNA sequence of DlUGD6 gene was 1 860 BP, including open reading frame 1443 BP, encoding 480 amino acids (GenBank accession number KU198438); Bioinformatics analysis showed that DlUGD6 was a stable acidic hydrophilic protein with no signal peptide, and had transmembrane structure and three typical conserved domains, belonging to the UDP- glucose / GDP- mannose dehydrogenase family. Phylogenetic tree analysis showed that DlUGD6 was closely related to citrus. (2) GFP fluorescence localization of onion inner epidermis showed that DlUGD6 was located in cytoplasm; QRT-PCR results showed that the expression of DlUGD6 was relatively high in non-embryogenic callus of longan and stable in other somatic embryogenesis. At the 8th (S3) and 24th (S7) days after the development of zygotic embryos, the highest expression level was observed, and the whole expression was "W" type. The expression of DlUGD6 in anthers and stems was the highest in different tissues and organs, and the overall expression level of DlUGD6 in reproductive organs was higher than that in vegetative organs. It is suggested that DlUGD6 gene may be involved in the synthesis of polysaccharides from the cell wall of longan in various stages of growth and development.
【作者单位】： 福建农林大学园艺植物生物工程研究所;
【基金】：国家自然科学基金(31572088,31272149) 福建省重大科技专项(2015NZ-0002-1)
【分类号】：Q943.2;S667.2
，
本文编号：2323924
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