摘要
9-顺式-环氧类胡萝卜素双加氧酶基因是脱落酸合成途径的关键基因。本试验通过RT—PCR结合RACE技术从苹果SH40（Malus domestica×Malus Honanensis）茎尖组织中克隆了1条NCED基因，命名为MdNCED1（GenBank登录号为KC816734）。该cDNA序列全长2179bp，包含1个编码606个氨基酸的开放阅读框。氨基酸同源性分析表明，MdNCED1与已报道的其他植物物种的氨基酸序列具有63．7％～93．0％的相似性。构建MdNCED1基因的原核表达载体pDEST15-MdNCED1，转入大肠杆菌（DE3），用IPTG诱导。SDS—PAGE分析表明，MdNCED1基因在大肠杆菌中被诱导表达的蛋白质分子量与预期结果一致。荧光定量结果表明，MdNCED1基因在SH28、M26、SH40及其嫁接品种嘎啦的表达趋势均呈先上升后下降的趋势，同时与其矮生程度呈正相关。
9-cis-epoxycarotenoid （NCED） gene is a critical gene in the pathway of abscisic acid biosynthesis. In this research,NCED gene in apple, designated as MdNCED1 （GenBank accession number： KC816734） ,was isolated from the apical tissue of apple rootstock SH40 （M. domestica x M. Honanensis） by RT-PCR and RACE technology. The full-length cDNA was 2179 bp, containing a complete open reading frame that encodes 606 amino acids. Amino acid sequence analysis revealed that the sequence had 63.7% -93.0% similarity with those of other reported plants. Construction of MdNCED1 gene prokaryotic expression vector pDEST15-MdNCED1 ,the recombined plasmid was transformed in to E. coli. （DE3） and induced by IPTG. The result of SDS-PAGE demonstrated that the size of expected protein in the prokaryotic expression system. Fluorescent quantitative PCR analysis showed that the trend of expression of SH28, M26, SH40, and the grafted varieties Gala were increased and then decreased. The expression of this gene was positively associated to the dwarfing degree.
出处
《植物遗传资源学报》
CAS
CSCD
北大核心
2014年第1期153-159,共7页
Journal of Plant Genetic Resources
基金
国家现代苹果产业技术体系(CARS-28)
