腾冲红花油茶1,6-二磷酸果糖醛缩酶基因克隆及组织表达分析  

Gene Cloning and Tissue Expression Analysis of 1,6-Diphosphate Aldolase in Camellia reticulata

作　　者：卿卓 苏睿[2] 赵文正 李林庶 任晓晓[1] 董坤 和绍禹[1] Qing Zhuo 1,Su Rui 2, Zhao Wenzheng 1, Li Linshu 3 ,Ren Xiaoxiao 1, Dong Kun 1, He Shaoyu 1(1 Faculty of Animal Science and Technology, Yurman Agricultural University, Kunming, 650201; 2 Sericulture and Apiculture Research Institute, Yunnan Academy of Agricultural Sciences, Mengzi, 661101; 3 Animal Husbandry Station, Tengchong, 67910)

机构地区：[1]云南农业大学动物科学技术学院,昆明650201 [2]云南省农业科学院蚕桑蜜蜂研究所,蒙自661101 [3]云南省腾冲市畜牧工作站,腾冲679100

出　　处：《分子植物育种》2018年第21期6982-6989,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31460577; 31572339);国家蜂产业技术体系项目(CARS-44-kxj13)共同资助

摘　　要：果糖-1,6-二磷酸醛缩酶(FBP2)不仅是植物糖酵解途径中核心调节酶之一,而且还能为油脂合成过程提供2种关键底物,对油料作物起着至关重要的作用。本研究以腾冲红花油茶为试材,根据已知普通油茶FBP2基因序列设计特异性引物,采用SMART RACE方法克隆出腾冲红花油茶1,6-二磷酸果糖醛缩酶全长cDNA,命名为CRFBPase (GenBank accession No. MG764086)。序列分析表明,该cDNA全长1 077 bp,编码358个氨基酸,分子质量为38 428.13 Da,等电点为7.56。根据cDNA序列推导出该蛋白氨基酸,利用DNAMAN与其他植物中的同源FBP2基因进行序列比对,发现腾冲红花油茶CRFBPase与普通油茶(JX914588.1)和拟南芥(BT000106.1) FBP2氨基酸的相似性分别为99.4%和66.9%。系统进化分析显示,腾冲红花油茶CRFBPase与普通油茶(JX914588.1)、马铃薯(DQ235169.1)、轮叶党参(AB243014.1) FBP2位于同一分支。组织表达分析结果显示,CRFBPase基因在腾冲红花油茶子房和种仁中均有一定量的表达且种仁中表达量最高。腾冲红花油茶和普通油茶种仁CRFBPase基因表达量与含油量的比较分析表明两者存在正相关关系。结果表明CRFBPase基因在腾冲红花油茶种仁发育调控过程大量诱导表达,且对其油脂代谢过程起着重要作用,为揭示腾冲红花油茶种仁中油脂代谢分子调控研究提供参考。Fructose-l,6-diphosphate aldolase （FBP2） in plants is not only one of the key regulatory enzyme in plant glycolysis pathway but also can provide two key substrate source for oil synthesis process, it plays an important role in oil crops. In this study, Camellia reticulata was used as the test material. According to the the known gene sequence of FBP2 common Camellia oleifera, specific primers were designed. The full-length cDNA of fructose-1,6-bisphosphate aldolase （FBP2） gene of Camellia reticulata seed was isolated and cloned by SMART R- ACE technology, and was designated as CRFBPase （GenBank accession No. MG764086）. The sequence analysis revealed that eDNA was a full length of 1 077 bp, encoding 358 amino acid with an estimated molecular weight of 38 428.13 Da and a calculated isoelectrie point （pI） of 7.56. According to the eDNA sequence, the amino acid ofthe protein was deduced, and then the sequence of the cDNA was carried out sequence alignment with the homologous FBP2 gene in other plants by DNAMAN. It was found that the similarities of FBP2 amino acids between Camellia reticulata CRFBPase and common camellia oleifera CoFBP2, Arabidopsis thaliana （BT000106.1） were 99.4% and 66.9%, respectively. Phylogenetic analysis showed that Camellia reticulata CRFBPase was in the same branch with CoFBP2, potato （DQ235169.1） FBP2 and Codonopsis lanceolatae （AB243014.1） FBP2. The results of tissue expression analysis showed that CRFBPase gene expressed in ovary and kernel of Camellia reticulata, and its expression in seed tissue were the highest. There was a positive relationship between expression of CRFBPase gene and oil content from seed in Camellia reticulata and Camellia oleifera. These results showed that CRFBPase gene might be induced and expressed in a large amount in the development process of seed, would play important roles in lipid metabolism in Camellia reticulata, and provide theoretical reference for the study of the molecular regulation of lipid metabolism of Camellia reticula

关 键 词：腾冲红花油茶 1 6-二磷酸果糖醛缩酶 基因表达 

分 类 号：S794.4[农业科学—林木遗传育种]