Abstract：A high frequency induced transformation system of hairy roots by Agrobacterium rhizogenes, that is an effective method to verify the gene function. Here, a genetic transformation system of Betula luminifera by A. rhizogenes was established, and had primarily optimized the culture condition of genetic transformation hairy root. First, 7 A. rhizogenes were used to infect B. luminifera leaves, and ArQual was obtained with the highest hairy root induction rate (69%). The optimal system for hairy root induction of B. luminifera was as follows: Leaves were pre-cultured in 1/2 MS medium for 2 days, infected for 20 minutes, and co-cultivation with acetosyringone (the concentration of 400 μmol/L) and cefotaxime (400 mg/L). In addition, the effect of different foreign gene vectors on hairy root induction was not significant. Based on the above system, pCAMBIA13011 vector containing β-glucosidase gene (GUS) gene and pGWB5 vector containing green fluorescent protein (GFP) gene were electroporated into ArQual, and transgenic hairy roots of B. luminifera were successfully induced with a conversion rate of 36.4%. In order to expand the application range of above genetic transformation system, the hypocotyls of B. luminifera seedlings (short cycle culture) were transformed by soaking and the stem segment of B. luminifera (long period culture) were punctured. After 30 days, the hairy roots were detected by GUS staining, fluorescence microscopy and PCR, and the results showed that GUS and GFP genes were successfully transformed and expressed, illustrated the different expression vector could express in new hairy roots of B. luminifera. In general, a high-frequency induction system for hairy roots of B. luminifera was established, which could be used to quickly verify gene function and expression in woody plants.
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