[Establishment of high efficiency genetic transformation system of maize mediated by Agrobacterium tumefaciens].
In order to establish high-frequency regeneration and high-efficiency genetic transformation system in maize, the significance of the 11 factors influencing maize embryonic callus induction and 9 factors affecting embryonic callus differentiation was researched by orthogonal experiment. The results showed that genotype had highly significant impact on induction of embryonic callus. The concentration of 6-BA, AgNO3, 2,4-D, ABA, and medium are the significant factors. The Multi-comparison showed that ABA 2 mg/L has a significant influence. Among the callus differentiation factors, the genotype and 6-BA concentration showed a strong main effect, the concentrations of NAA, medium, KT and 2,4-D had significant impacts on callus differentiation. Southern blotting analysis demonstrated that the resistant callus rate under the selection pressure of 25 mg/L hygromycin was a reliable indicator for system optimization in resistance screening. The concentration of acetosyringone (AS) showed sensitive differences among genotypes. The highest transformation rate was found with the optimized combination of 24-25 degrees C for co-culture temperature, 0.7 ODx15 min for Agrobacterium tumefa-ciens concentration and incubation-time, and pH 5.5-6.2. By this optimized combination, the survival rate of resistant calli as an index for the stable transformation rates of inbred lines Huangzao 4 and Zong 31 by introducing GUS gene into maize inbred lines was as high as 48.6% and 46.2%, respectively.
WEI
Kai-Fa
KF
Department of Biological Sciences and Biotechnology, Zhangzhou Normal University, Zhangzhou 363000, China. kaifa-wei@163.com
D000040:Abscisic Acid / Q000494:pharmacology; D016960:Agrobacterium tumefaciens / Q000235:genetics*; D001593:Benzyl Compounds; D015139:Blotting, Southern; D006863:Hydrogen-Ion Concentration; D007701:Kinetin / Q000494:pharmacology; D008957:Models, Genetic; D011687:Purines; D012835:Silver Nitrate / Q000494:pharmacology; D014170:Transformation, Genetic / Q000187:drug effects / Q000235:genetics*; D003313:Zea mays / Q000235:genetics*
D001593:Benzyl Compounds; D011687:Purines; D000040:Abscisic Acid; D012835:Silver Nitrate; C480551:benzylaminopurine; D007701:Kinetin