摘要
为研究花青素苷的转运,利用电子克隆和RT-PCR方法,从普通烟草(Nicotiana tabacum)花中分离了1个编码谷胱甘肽转移酶(GST)基因,命名为NtAN9(GenBank登录号KX356542)。NtAN9包含一个690bp的开放阅读框,编码229个氨基酸残基,属于phi型GST。NtAN9基因组结构由3个外显子和2个内含子组成。多序列比对分析表明,NtAN9与矮牵牛(Petunia hybrida)花青素苷转运相关的GST基因PhAN9具有88%的一致性。系统进化分析显示,NtAN9与花青素苷转运相关的GST基因聚为一支,是PhAN9的直系同源基因。定量PCR分析表明,NtAN9基因在含有花青素苷的四个花发育时期中均有表达,其中在开花前的第Ⅲ期(2cm<花芽<4cm)表达丰度达到最高,而在不含花青素苷的根、茎和叶中不表达。由此推测,分离得到的NtAN9可能具有类似PhAN9的功能,与烟草花青素苷的转运与积累相关。NtAN9基因的分离与表达分析,为进一步研究烟草花青素苷的转运奠定了基础。
To study anthocyanin transport,we isolated a full-length gene NtA N9(GenBank Accession No.KX356542)by in silicon cloning and RT-PCR method from Nicotiana tabacum.The NtA N9 contains a 690 bp open reading frame(ORF)encoding 229 amino acid residues,which belongs to a putative phi class glutathiones-transferase(GST)family.Itsgenomicstructurecontains3exonsand2introns.Multiplese-quence alignment showed that NtAN9 has highest identities(88%)with petunia PhAN9 that is an antho-cyanin transport-related GST.Phylogenetic analysis showed that NtA N9 was clustered into an anthocya-nin transport-related GST group,and was an ortholog of PhA N11.The anthocyanin content analysis and real-time quantitation PCR(qRT-PCR)showed that NtA N9 was expressed in different flower develop-mental stages,with highest level in stageⅢ,but almost no expression in root,stem and leaf w hich are al-most no anthocyanin.These results suggest that the NtA N9 function is similar with PhA N9 and might be related to the transfer and accumulation of anthocyanin in tobacco plants.
作者
崔超军
李嘉敏
陈君宇
张润钊
祝钦泷
CUI Chaojun;LI Jiamin;CHEN Junyu;ZHANG Runzhao;ZHU Qinlong(College of Life Sciences,South China Agricultural University,State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Key Laboratory of Plant Functional Genomics and Biotechnology,Education Department of Guangdong Province,Guangzhou 510642 China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2016年第7期1337-1342,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家级大学生创新创业训练计划项目(201610564006)
广东省公益研究与能力建设项目(2016A020210084)
关键词
烟草
花青素苷转运
谷胱甘肽转移酶
基因克隆
表达分析
N icotiana tabacum
anthocyanin transport
glutathione s-transferase
gene clone
expression analy sis
