Optimization and Application of Transient Transformation System of Larix kaempferi
[Objective]To optimize an Agrobacterium-mediated transient transformation system with Larix kaempferi embryogenic callus.[Methods]The embryogenic callus of Larix kaempferi cultured in liquid me-dium for 7 days was used as the receptor material,and pCAMBIA1305.1 vector carrying β-glucuronidase(GUS)was used for transient transformation.Based on the expression level and enzyme activity of GUS,the optimal infection solution concentration,infection time and co-culture time were screened.The activity of Larix kaempferi scarecrow-like 6(LaSCL6)promoter was analyzed with the screened transformation system.[Results]After transient transformation,the expression of GUS was obvious.When the concentra-tion of infection solution was 0.2,the infection lasted for 5 minutes,and the co-culture time was 72 hours,GUS expression was the highest,with-2.2742.When the concentration of infection solution was 0.05,the infection lasted for 5 minutes,and the co-culture time was 72 hours,GUS enzyme activity was the highest with 25.7286 U/L.The activity of LaSCL6 promoter was 1.55 times higher than that of CaMV35S pro-moter[Conclusion]In view of the expression level and enzyme activity of GUS,transformation efficiency is high when the concentration of infection solution is 0.05,the infection time is 5 minutes,and the co-culture time is 24 hours,which can be used for efficient transformation of embryogenic callus of Larix kaempferi.
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