文献类型：期刊文献

中文题名：矮生山茶‘恨天高’GA20ox1的克隆及其对转基因烟草株形的影响

英文题名：Overexpression and Silence Expression of CrGA20ox1 from Camellia reticulata ‘Hentiangao’ and Its Effect on Plant Forms in Transgenic Tobacco

作者：王江英[1] 吴斌[1] 刘伟鑫[1] 范正琪[1] 李纪元[1]

第一作者：王江英

机构：[1]中国林业科学研究院亚热带林业研究所,浙江省林木育种技术研究重点实验室,浙江富阳311400

年份：2015

卷号：42

期号：8

起止页码：1523-1532

中文期刊名：园艺学报

外文期刊名：Acta Horticulturae Sinica

收录：CSTPCD;;Scopus;北大核心:【北大核心2014】；CSCD:【CSCD2015_2016】；

基金：国家‘十二五’科技计划课题(2012BA01B0703);浙江省花卉新品种选育重大科技专项(2012C12909-6);浙江省与中国林业科学研究院省院合作林业科技项目(2012SY02);国家农业成果转化资金项目(GB2013GB24320606);引进国际先进林业科学技术项目(948项目)(2014-4-16)

语种：中文

中文关键词：山茶;云南山茶;矮生;基因克隆;表达分析;转基因烟草

外文关键词：Camellia japonica; Camellia reticulata; dwarfing variety; gene clone; expression analysis; transgenic tobacco

分类号：S685.14

摘要：根据山茶(Camellia japonica)同源序列设计特异性引物,利用同源克隆和3′,5′-RACE技术,从云南山茶(C.reticulata)矮生品种‘恨天高’茎尖组织中克隆出GA20–氧化酶(GA20-oxidase,GA20ox)基因,命名为Cr GA20ox1(Gen Bank登录号:KP635268),基因全长1 178 bp,开放阅读框1 146 bp,编码381个氨基酸。实时荧光定量PCR分析发现,GA20ox1在4种不同的山茶中均得到表达,其中乔木类的浙江红山茶(C.chekiangoleosa)GA20ox1表达量最高,其次是岳麓连蕊茶(C.handelii)及金花茶(C.nitidissima),表达量最低的为矮生品种‘恨天高’。结果表明该基因的表达量高低与测试物种的株高有关。GA20ox1在4个物种不同组织中的表达模式基本一致,且在茎尖组织中的表达量均高于叶片。Cr GA20ox1转基因烟草分析结果表明,过表达Cr GA20ox1烟草株高约为野生型的3.6倍,而沉默表达Cr GA20ox1烟草株高约为野生型的一半。
On the basic of homologous sequences of Camellia japonica,a GA20-oxdiase（GA20ox）gene was isolated from the stem tip of C. reticulata‘Hentiangao＇by the 3′,5′-RACE technology named Cr GA20ox1（Gen Bank Accession ID：KP635268）. The full-length c DNA of Cr GA20ox1 is 1 178 bp,containing an 1 146 bp ORF which encodes 381 amino acids. Expressions of GA20ox1 in four different plant phenotypes of camellias were analyzed by fluorescent quantitative real-time PCR. The results showed that GA20ox1 gene was expressed in the four camellias differently,where the expression level of GA20ox1 in C. chekiangoleosa was the highest,medium in C. handelii and C. nitidissima,and the lowest in C. reticulata‘Hentiangao＇,which indicated that the expression intensity of the GA20ox1 gene was positively associated with height in the different camellias. The trends of expressions of GA20ox1 gene were almost the same in various organs of the four camellias,and furthermore,the expressions of the gene was higher in the shoot tips than in tender leaves. Phenotypic results of transgenic tobacco showed that the transgenic plant with sense Cr GA20ox1 gene was nearly 3.6 times height than wild type tobacco;Meanwhile,the transgenic plant with anti-sense Cr GA20ox1 gene was about only half height of the wild type tobacco.