摘要
以龙眼胚性愈伤组织为材料,采用RT-PCR和RACE-PCR技术对植物抗寒重要调控因子HOS1(DlHOS1)进行克隆,并进行密码子使用偏爱性、生物信息学和低温胁迫下表达等方面的分析。结果表明:DlHOS1基因序列全长3 577 bp,包含ORF 3 000 bp,编码999个氨基酸,5’UTR和3’UTR分别162 bp和415 bp(GenBank登录号:KY990404);密码子使用偏爱性分析表明,DlHOS1密码子的选择偏爱性较弱,偏爱以A/T结尾;生物信息学分析结果表明,DlHOS1编码的蛋白属于不稳定的疏水性跨膜蛋白,不含信号肽,定位于细胞质和细胞核,二级结构富含α螺旋,与柑橘的亲缘关系较近;qPCR结果发现,DlHOS1能够响应低温胁迫诱导,总体上低温下其表达水平上升。研究结果认为,DlHOS1参与龙眼抗寒和低温胁迫过程。
To study the function of HOS1 in cold stress of Dimocarpus longan Lour., the embryogenic callus of D. longan Lour were used to clone HOS1 gene （DIHOS1） , and codon bias, bioinformatics and expression pattern of DIHOS1 were analyzed. The results showed that the full length sequence of DlHOS1 was 3 577 bp, containing an ORF 3 000 bp, encoding 999 amino acids, and the 5＇ UTR and 3＇ UTR was 162 bp and 415 bp respectively （GenBank accession number： KY990d04）. The codon bias level of DIHOS1 was low, and it biased toward the synonymous codons with A and T at the third codon position. Bioinformatics analysis indicated that DIHOS1 was an unstable and hydrophobic transmembrane protein, had no signal peptide and multilocated in cytoplasm and nucleus. The secondary structure was mainly composed of alpha helix and strand. Besides, D1HOS1 shared a very close relationship with citrus, qPCR analysis showed that DIHOS1 was response to the induction of low temperature and up-regulated at low temperature. The research suggested that DIHOS1 should be involved in cold resistance and low temperature stress in longan.
作者
肖小娥
徐小萍
李佳蜜
张梓浩
赖瑞联
林玉玲
赖钟雄
XIAO Xiaoe1, XU Xiaoping1, LI Jiami1, ZHANG Zihao1, LAI Ruilian1,2, LIN Yuling1 , LAI Zhongxiong1(1 Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China 2 Fruit Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013, Chin)
出处
《热带作物学报》
CSCD
北大核心
2018年第2期293-299,共7页
Chinese Journal of Tropical Crops
基金
国家自然科学基金(No.31572088
No.31672127)
福建省重大科技专项(No.2015NZ0002-1)
关键词
龙眼
DlHOS1
基因克隆
密码子使用偏爱性
qPCR
Dimocarpus longan Lour., DIHOS1
gene cloning
codon bias
Real-time fluorescent quantitative PCR
