Agrobacterium-mediated in planta transformation of maize germ cells | Агробактериальная трансформация генеративных клеток кукурузы in planta
2010
Velikov, V.A. | Mamontova, E.A. | Volokhina, I.V., Russian Academy of Sciences, Saratov (Russian Federation). Institute of Biochemistry and Physiology of Plants and Microorganisms | Chumakov, M.I., The N.G. Chernyshevskij Saratov State Univ. Balashov Branch (Russian Federation)
A method of maize (Zea mays (L.) Moench) genetic translation based on the in planta agrobacterial transformation of vering plants has been tested using two maternal and two paternal maize genotypes. Transformation was performed with the 3 of constructs containing the npt2 and (beta-glucuronidase gus-intron, and gus) marker and reporter genes. The presence of the marker and reporter genes in the DNA of seedlings grown from the seeds exposed to the treatment was determined by PCR and by histochemical analysis. The efficiency of the marker genes insertion in the genomes of two maize diploid lines (Korichnevyj Marker Saratovskij and AT-3) after fertilization with pollen from the lines of Zarodyshevyi Marker Saratovskii and Zarodyshevyi Marker Saratovskij Purpurnyj, respectively, was found out. For AT-3 maize line transformed with the pTd33 and pA2069 constructs, the efficiency was 13% and 5,4% of the total number of obtained seeds, respectively. For Korichnevyj Marker Saratovskij line transformed with the above constructs the efficiency values were 12,1% and 13,9%, respectively. Overall, the effectiveness of the method was greater than that of the traditional techniques for agrobacterial transformation via vegetative cells, regardless of transformed line and transferred construction. The method can be used to create novel maize varieties and hybrids, particularly in transformation of lines with poor regeneration ability
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