干旱区生态环境知识资源中心(Arid): 沙冬青AmFAD2-1基因的克隆及其功能分析

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	沙冬青AmFAD2-1基因的克隆及其功能分析
其他题名	Cloning and Functional Analysis of AmFAD2-1Gene fromAmmopiptanthus mongolicus
	张文君; 薛敏; 任美艳; 唐宽刚; 张宇; 王茅雁
来源期刊	西北植物学报
ISSN	1000-4025
出版年	2019
卷号	39 期号:11 页码:1901-1910
中文摘要	细胞膜脂的不饱和度主要由脂肪酸去饱和酶(fatty acid desaturase,FAD)决定,二者与植物抵抗低温和高盐等逆境胁迫密切相关。沙冬青(Ammopiptanthus mongolicus)是中亚荒漠区唯一的常绿旱生阔叶植物,具有很强的耐寒、耐旱和耐盐碱等特性。该研究利用RT-PCR方法克隆到沙冬青油酸去饱和酶基因AmFAD2-1的cDNA和gDNA,其中后者不含内含子。在AmFAD2-1蛋白(含382个氨基酸残基)序列中含有FAD家族必须的保守组氨酸簇和内质网定位信号及跨膜结构域。进化分析表明,AmFAD2-1与大豆(Glycine max)和柠条锦鸡儿(Caragana korshinskii)等豆科植物的FAD2距离较近,而与玉米(Zea mays)和拟南芥(Arabidopsis thaliana)等其他植物的FAD2距离较远。qRT-PCR分析表明,在室内培养的沙冬青幼苗中,AmFAD2-1的表达量在低温处理2~24h和干旱处理6~13d期间显著上调,在盐胁迫处理期间则呈现升高、降低、再升高的变化趋势;在野外生长的沙冬青成株嫩叶中,其表达量在秋季和冬季总体上明显高于春季和夏季。成功构建了植物表达载体p3300-35S-Am- FAD2-1并通过根癌农杆菌介导法转化野生型拟南芥,获得16株转AmFAD2-1基因植株。耐逆性鉴定表明,转基因株系的耐冻性较野生型拟南芥显著提高,其耐旱性和耐盐性也比后者有较明显的改善。
英文摘要	The unsaturation degree of cellular membrane lipids is mainly determined by fatty acid desaturases(FADs),and both the unsaturation degree and FADs are closely related to the tolerance of plants to harsh environments such as low temperature and salt stress.Ammopiptanthus mongolicus is the only evergreen xerophytic broadleaf plant that is endemic to the central Asian desert and has very strong tolerance to abiotic stresses such as cold,drought and high salinity.In this study,both the cDNA and gDNA fragments for an oleic acid desaturase,AmFAD2-1,in this species were cloned by RT-PCR,and the gDNA region has no intron.Except for the conserved histidine clusters for FAD family,there also exist the signal peptide for endoplasmic reticulum localization and transmembrane domains in the AmFAD2-1sequence (containing 382amino acid residues).Phylogenetic relationship analysis showed that AmFAD2-1is closely related to the FAD2sfrom leguminous plants such as Glycine maxand Caragana korshinskii but is distantly related to the FAD2sfrom other plant species such as Zea mays and Arabidopsis thaliana.qRT-PCR analysis showed that in laboratory-cultured A.mongolicus seedlings,the expression level of AmFAD2-1 increased significantly between 2to 24hof the cold treatment and between 6to 13dof the drought treatment. Moreover,AmFAD2-1showed an expression pattern of increasing,decreasing and increasing again during the period of salt treatment.In young leaves of the A.mongolicus plants naturally growing in the wild,the expression levels of AmFAD2-1during the autumn and winter months were overall obviously higher than those in the spring and summer.The plant expression vector p3300-35S-AmFAD2-1was successfully constructed and wild type Arabidopsis was transformed using the Agrobacterium tumefaciensmediated method.In total,16 AmFAD2-1transgenic plants were obtained.Stress tolerance evaluation experiments revealed that the transgenic lines had significantly higher freezing tolerance compared with the wild type Arabidopsis;their drought and salt tolerances were also improved.
中文关键词	沙冬青 ; 基因表达 ; 转基因 ; 耐逆性
英文关键词	FAD2 Ammopiptanthus mongolicus FAD2 gene expression transgene stress tolerance
语种	中文
收录类别	CSCD
WOS类目	GENETICS HEREDITY
WOS研究方向	Genetics & Heredity
CSCD记录号	CSCD:6625091
来源机构	内蒙古农业大学
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/315986
作者单位	内蒙古农业大学生命科学学院, 呼和浩特, 内蒙古 010018, 中国
推荐引用方式 GB/T 7714	张文君,薛敏,任美艳,等. 沙冬青AmFAD2-1基因的克隆及其功能分析[J]. 内蒙古农业大学,2019,39(11):1901-1910.
APA	张文君,薛敏,任美艳,唐宽刚,张宇,&王茅雁.(2019).沙冬青AmFAD2-1基因的克隆及其功能分析.西北植物学报,39(11),1901-1910.
MLA	张文君,et al."沙冬青AmFAD2-1基因的克隆及其功能分析".西北植物学报 39.11(2019):1901-1910.