中国水仙NtPI2基因的克隆及遗传转化  

Clone and Genetic Transformation of NtPI2 Gene from Narcissus tazetta var.chinensis

作　　者：吴菁华[1] 武丹[1] 吴少华[1] 

机构地区：[1]福建农林大学园艺学院,福州350002

出　　处：《西北植物学报》2017年第6期1059-1065,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(31601783);福建省自然科学基金(2014J01083)

摘　　要：采用RT-PCR和RACE技术,从中国水仙‘金盏银台’中获得1个MADS-box基因NtPI2。NtPI2基因全长810bp,含有1个627bp开放阅读框,编码208个氨基酸。系统进化树显示,NtPI2属于B类MADS-box基因家族的PI/GLO类基因。荧光定量PCR分析表明,NtPI2基因在中国水仙‘金盏银台’各营养器官中都有表达;在单瓣和重瓣水仙花朵的各个部位均有表达,但表达模式存在差异。比较NtPI2在单瓣和重瓣中国水仙中的表达模式,发现重瓣水仙‘玉玲珑’的瓣化雄蕊和副冠的NtPI2基因表达量较单瓣水仙‘金盏银台’显著增高,推测NtPI2基因在瓣化雄蕊中表达量显著增高可能是重瓣中国水仙发生的直接原因。成功构建了NtPI2基因的pCAMBIA1302-NtPI2超表达载体,并通过农杆菌介导法进行烟草的遗传转化,分子鉴定结果表明共获得了8株转NtPI2基因植株。本研究为深入探索NtPI2基因的功能及其与中国水仙重瓣花形成的关系奠定了基础。A MADS-box gene, NtPI2, was cloned from the flower bud of Narcissus tazetta var.chinensis using RT-PCR and rapid amplification of cDNA ends.NtPI2 is 810 bp in length with an open reading frame 627 bp which encoded 208 amino acids.Phylogenetic analysis indicated that NtPI2 belongs to the PI/GLO family.Real-time quantitative PCR demonstrated that NtPI2 was expressed in all tissues of ＆#39;Jinzhanyintai＆#39;.The expression pattern of NtPI2 was different in different organs of flower in single and double narcissus.We compared the expression pattern of NtPI2 in single and double narcissus, and found that the expression of the NtPI2 of the stamen petaloids and corona in double narcissus ＆#39;Yu-linglong＆#39; were significantly higher than that of in single narcissus ＆#39;Jinzhanyintai＆#39;.This indicates that the increased expression of NtPI2 in the petaloid stamens may be the direct cause of the occurrence of the double narcissus.Plant express vector of NtPI2, pCAMBIA1302-NtPI2, was constructed and used to transform the tobacco through Agrobacterium-mediated procedure.The result of molecular identification showed that 8 transgenic tobacco plants were obtained.The results will lay a foundation for the further study of the NtPI2 gene function and its relationship with the formation of the double narcissus.

关 键 词：中国水仙 花器官发育 B类MADS-box基因 NtPI2基因 

分 类 号：Q785[生物学—分子生物学] Q786