根癌农杆菌介导的反义VeFAD2基因对少根根霉的遗传转化及其影响因子

Agrobacteriurn tumefaciens-mediated transformation (AtMT) method was used to study the genetic transformation in Rhizopus arrhizus of antisense expression vector pBI121-fad2 of VeFAD₂ gene. We studied the key factors influencing transformation, such as A.tumefaciens strains,bacterial cell volume initially used, co-cultivation time, effective period and concentration of AS. The results shows that the ideal A.tumefaciens strain is AGL-1, the optimum bacterial cell volume initially used is 50-100 μL and co-culture 24 hours will get the most transformants. At co-culture period, AS induction is indispensable. It is beneficial to improve transformation ratio by adding to 200 μmol·L^-1 AS at preincubate period and increasing AS concentration to 400-600 μmol·L^-1 at co-culture period. It determined that antisense VeFAD₂ gene has been integrated into the genome of R. arrhizus by PCR-Southern detection.