Expression Of Transformed Wheat With Anti-PLDγ Gene And The Preliminary Study Of Male Sterile Mutant

Antisense phospholipase Dγ(PLDγ) gene was introduced into LK906 wheat mediated by improved earstem injecting of Agrobacterium. The study has detected the anti-PLDγ. The transformants were screened by PCR and RT-PCR technique based on Kanamycin selection. The result showed anti- PLDγgene was integrated into wheat genomes and expressed successfully. The variance between transgenic lines and receptor on the Chlorophyll content and Pn were studied. At the same time, the male sterile mutant in the transformed wheat was studied preliminary. The main results were as follows:1. The expression of anti-PLDγgene in the transgenic wheat was proved firstly. Three transgenic plants were obtained by canamycin selection, PCR and RT-PCR amplification. The special fragments of 500bp and 195bp, which were consistent with the plasmid, had been detected in these transgenic plants.2. The mutant plants, 03039S, could be identified easily because of its dwarfism and distinct sterility traits include the light green plans and spikes, the expanding spikes, thin and not opening anthers. The result that the rate of abnormity frequency of microspore was 86.12% showed that the sterility mainly took place in monocyte stage. The abnormity turned more aggressive in the tricellular. The female fertility was normal after hybridizing with different materials, compared with the host plants. The separate proportion of of fertility and sterility was 1:1.3. The mutants of agronomic traits were induced in 05090RT by the antisense phospholipase Dγin the plant height, 1000-kernal weight, spikes per plant. Positive influences were showed in this three traits. Compared with the transgenic plants, the average height of 05090RT was higher than the host plants by 15.7 cm, 7.48g in the 1000-kernal weight, and 2.5 in spikes per plant. However, the number of spikelets and grains per spike were lower than the transgenic plants, 4.0 and 8.7 lower respectively compared with the host plants. The mature period of transgenic plants was shorter than the host plants, which would do good to improve the disadvantage of later mature period. 4. The transgenic plants, 05090RT, had no significant difference with host plants in Chlorophyll content and Pn. The dynamics of Chlorophyll content were little higher than the host plants during the different stages, heading stage, anthesis stage, mid-filling stage and later filling stage. The range was between 1.43 and 9.37, but the T-value wasn't significant. As to the Pn and Gs, same effect was reflected. The difference of Tr between the transgenic plants and host plants was significant in heading stage, up to 0.24μmolH2O ?m-2?s-1, however, the difference was not significant during other stages. The results showed that the antisense phospholipase Dγhad no evident effect on Chlorophyll content and Pn of the transgenic plants.