Method for transient expression of introducing exogenous gene into chrysanthemum or related species

Abstract

The invention provides a method for transient expression of introducing an exogenous gene into chrysanthemum or related species. The exogenous gene is a GUS gene which is carried by an over-expression vector plasmid of smaller than 10 kb carrying a 35s promoter and a terminator, and the method mainly comprises the following steps: (1) obtaining of a plant; (2) building of a plant expression vector pORER1-2X35s; (3) preparation of an infection liquid; (4) preparation of chrysanthemum leaves; (5) transformation of leaves; (6) cultivation of leaves after transformation; and (7) detection of positive transformed leaves. According to the method, appropriate explants are selected, agrobacterium tumefaciens strains serve as the susceptible strains, and infection thalli are selected through rifampicin and kanamycin; simultaneously P19 and agrobacterium tumefaciens are mixed and cultivated to be prepared into the infection liquid, P19 can restrain the silence effect of plants to vector expression exogenously introduced, so that the expression level is improved, the co-culture conditions are reasonably optimized, transformation time based on the method is shortened to about 45 days, and the transformation rate reaches 60 percent above; the method of the invention provides possibility for effective screening and analysis of chrysanthemum gene function, protein positioning and further obtaining of stable transformants.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a method for transiently expressing exogenous genes into chrysanthemums or related species through the injection infiltration method of agrobacterium. Background technique [0002] Chrysanthemum (Chrysanthemum morifolium Ramat.), native to China, belongs to the perennial herbaceous flowers of the genus Chrysanthemum in the family Asteraceae. It has a cultivation history of more than 3,000 years in my country and is one of the top ten famous flowers in China. 30% of the world's cut flowers (Hong Bo et al., 2009). Chrysanthemum has high ornamental value and has been used more and more in landscaping and beautification in recent years. It is a kind of mass consumption flower, and it is urgent to increase the progress of chrysanthemum variety breeding and innovation work (Zhu Mingtao et al., 2011), so as to provide good economic benefits and social benefits. ...

AI Technical Summary

Problems solved by technology

[0005] The invention provides a method for transiently expressing foreign genes into chrysanthemums and related species. Mediated by Agrobacterium, the plant expression vector pORER1-2×35s:gus is constructed so that the foreign gene GUS can be expressed in chrysanthemums and chrysanthemums. Transient expression of related species solves the problem of long transformation period and low efficiency of Agrobacterium-transformed chrysanthemum genetic transformation, and provides the possibility for more effective screening and analysis of chrysanthemum gene function, protein localization and further obtaining stable transformants

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