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Transient genetic transformation of plant organs is an indispensable way of studying gene function in plants. This study was
aimed to develop an optimized system for transient Agrobacterium-mediated transformation of the Arabidopsis leaves. The β-glucuronidase (GUS) reporter gene was employed to evaluate growth and biochemical parameters that influence
the levels of transient expression. The effects of plant culture conditions, Agrobacterial genetic backgrounds, densities of Agrobacterial cell suspensions, and of several detergents were analyzed. We found that optimization of plant culture conditions is the
most critical factor among the parameters analyzed. Higher levels of transient expression were observed in plants grown under
short day conditions (SDs) than in plants grown under long day conditions (LDs). Furthermore, incubation of the plants under
SDs at high relative humidity (85–90%) for 24 h after infiltration greatly improved the levels of transient expression. Under
the optimized culture conditions, expression of the reporter gene reached the peak 3 days after infiltration and was rapidly
decreased after the peak. Among the five Agrobacterial strains examined, LAB4404 produced the highest levels of expression. We also examined the effects of detergents, including
Triton X-100, Tween-20, and Silwet L-77. Supplementation of the infiltration media either with 0.01% Triton X-100 or 0.01%
Tween-20 improved the levels of expression by approximately 1.6-fold. Our observations indicate that transient transformation
of the Arabidopsis leaves in the infiltration media supplemented with 0.01% Triton X-100 and incubation of the infiltrated plants under SDs
at high relative humidity are necessary for maximal levels of expression. 相似文献