Transgenic wheat progeny resistant to powdery mildew generated by Agrobacterium inoculum to the basal portion of wheat seedling.
To improve the transformation efficiency of wheat (Triticum aestivum L.) mediated by Agrobacterium tumefaciens, we explored the possibility of employing the basal portion of wheat seedling (shoot apical meristem) as the explants. Three genotypes of wheat were transformed by A. tumefaciens carrying beta-1, 3-glucanase gene. After vernalization, the seeds to be transformed were germinated. When these seedlings grew up to 2 approximately 5 cm, their coleoptile and half of the cotyledon were cut out, and the basal portions were infected by A. tumefaciens. A total 27 T(0) transgenic plants were obtained, and the average transformation efficiency was as high as 9.82%. Evident segregation occurred in some of the T(1) plants, as was indicated by PCR and Southern blotting analysis. Investigation of the T(2) plants revealed that some transformed plants had higher resistance to powdery mildew than the controls. Northern blotting revealed that beta-1, 3-glucanase gene was normally expressed in the T(2) plants, which showed an increased resistance to powdery mildew. The results above indicate that the exogenous gene has been successfully integrated into the genome of wheat, transmitted and expressed in the transgenic progeny. From all the results above, it can be concluded that Agrobacterium inoculum to the basal portion of wheat seedling is a highly efficient and dependable transformation method. It can be developed into a practicable method for transfer of target gene into wheat.
Zhao
Tong-Jin
TJ
School of Life Sciences, Shandong University, Jinan, 250100, PR China.
Zhao
Shuang-Yi
SY
Chen
Hui-Min
HM
Zhao
Qing-Zhen
QZ
Hu
Zan-Min
ZM
Hou
Bing-Kai
BK
Xia
Guang-Min
GM
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D001203:Ascomycota; D017343:Genes, Plant; D043326:Glucan 1,3-beta-Glucosidase / Q000235:genetics / Q000378:metabolism; D010935:Plant Diseases; D030821:Plants, Genetically Modified / Q000382:microbiology*; D036226:Seedlings / Q000235:genetics / Q000382:microbiology; D014170:Transformation, Genetic; D014908:Triticum / Q000235:genetics* / Q000382:microbiology
D043326:Glucan 1,3-beta-Glucosidase