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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 protein widely exists in eukaryotes and assists cells to perform a variety of functions in vivo. At present, the research on WD40 protein is mostly concentrated in Arabidopsis and rice.Bioinformatics analysis shows that SlWD6 protein contains two conserved WD -repeat domain belongs to the WD40 family.In order to understand the function of SlWD6 gene in tomato, the expression of SlWD6 gene in tomato root, stem, leaf, flower and fruit at different developmental stages was detected by RT-PCR.The RT-PCR method was used to obtain SlWD6 gene was constructed. The overexpression vector of SlWD6 was constructed and transgenic plants were obtained by Agrobacterium tumefaciens-mediated method. Realtime PCR was used to detect the expression of SlWD6 gene in three independent transgenic lines (WD6-393, WD6-418 and WD6-421) The results showed that the expression of SlWD6 gene in tomato was constitutively expressed and the expression of SlWD6 gene was highest in the red fruit during the whole fruit period. The expression of SlWD6 gene was significantly higher in the transgenic lines In wild type, under drought and salt stress, the content of Pro in leaves of transgenic plants was significantly higher than that in wild type, while the content of malondialdehyde (MDA) in wild type was significantly lower than that of wild type Mannitol mediated salt tolerance and drought stress, and the root length and seedling length of T2 generation of S1WD6 transgenic plants were significantly higher than that of wild type plants.In summary, overexpression of SlWD6 gene could significantly increase the drought resistance and salt tolerance of tomato.