摘要
WD40蛋白广泛存在于真核生物体内,在生物体内协助细胞行使多种功能,目前关于WD40蛋白的研究多集中在拟南芥菜和水稻中.生物信息学分析显示,SlWD6蛋白包含两个保守的WD-repeat结构域,属于WD40家族.为了解番茄中SlWD6基因的功能,采用RT-PCR方法检测番茄的根、茎、叶、花和不同发育时期果实中SlWD6基因表达量.利用RT-PCR方法获得SlWD6基因全长,并且构建SlWD6过量表达载体,通过农杆菌介导法获得转基因植株,利用Realtime PCR检测3个独立的转基因株系(WD6-393、WD6-418和WD6-421)中SlWD6基因的表达量,并进行耐盐和抗旱性分析.结果显示,番茄SlWD6基因为组成型表达,果实各时期表达量较高,在红果时期表达量达到最高;转基因株系中SlWD6基因的表达量显著高于野生型;在干旱和高盐胁迫下,转基因植株叶片脯氨酸(Pro)含量显著高于野生型,丙二醛(MDA)含量与野生型相比则显著降低.用Na Cl和甘露醇介导耐盐和干旱胁迫,SlWD6转基因植株T2代种子的根长和苗长显著高于野生型植株.综上,SlWD6基因的过量表达能够显著增强番茄的抗旱和耐盐功能.
WD40 proteins widely existent in eukaryotic organisms are important to cell functions and therefore well studied in Arabidopsis and Rice. However, little information is available about the proteins in tomato. Bioinformatics analysis showed that S1WD6 protein has two WD-repeat conserved domains, therefore belonging to the family of WD40 Protein. In order to explore the function of WD40 proteins in tomato, we cloned the full length of SlWD6 gene and detected the SIWD6 gene expression level of the roots, stems, leaves, flowers and different stages of fruits in tomatoes by RT-PCR. Transgenic plants over-expressing SlWD6 were obtained by Agrobacterium mediated transformation. Real-time PCR was used for the detection of SIWD6 gene expression, and three independent transgenic lines （WD6-393, WD6-418, WD6-421） were selected for salt and drought stress analysis. The results suggested that the SIWD6 gene belongs to constitutive expression in tomato, and that the gene expression level is higher in fruit maturation than in other organs, being the highest at the red fruit stage. The expression level of SIWD6 was significantly up-regulated in transgenic lines compared with wild type. The content of proline （Pro） was increased and malonic dialdehyde （MDA） was reduced as compared with the wild type in drought and salt stress growth. The root length and seedling length of transgenic plants are both higher than the wild type under NaC1 or mannitol treatment. In addition, over-expressing SlWD6 gene can significantly improve drought and salt tolerance of tomato.
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2015年第3期413-420,共8页
Chinese Journal of Applied and Environmental Biology
基金
国家杰出青年科学基金项目(30825030)
国家自然科学基金项目(31171179
90717110)
国家973计划项目(2011CB100401)资助~~
