| The use of modified spikes injection by transfering antisense- PLDγgene into maize inbred lines Zheng 58, L9801 via Agrobacterium–mediated . On the basis of kanamycin selection, PCR amplified and PCR-Southern hybridization, the results proved that antisense-PLDγgenes has integrated into the chromosome genome of the receptor. Meanwhile by the analysis of transgenic lines offspring variation, salt and drought tolerance, the main agronomic traits difference, and biological software analysis of transgenic fragment , the results are as follows:1.It is the first time to obtain the transgenic corn plants of antisense- PLDγgenes. Kanamycin resistance plants PCR amplified ,12 materials were amplified the same size of the fragments as plasmid was done. PCR amplification and PCR-Southern hybridization are consistent. It is proved that the gene has been integrated into maize genome and the transgenic corn plants were obtained.2. The male sterile plants were found in T0, T1 generation. Many male-sterile corn variation strains have been found in the transgenic plants of the different generations. The plants are relatively short stature, slow growth, poor development-Picayune, and flowers distributed more sparsely, not shed lately and stamen remains in Ying crust after ripe. CMS lines and the control have the notable difference in the height, the length of Picayune, Picayune– branches and the outcome, the height difference was particularly marked, ear height is only slightly different.3. The aging physiological indicators analysis of T1 generation transgenic corn. It is found that SOD, POD, soluble proteins and CAT in the total content of transgenic plants and the control have no significant changes, the only change is a change in date, the date of aging is delayed about 6-10 days. MDA and chlorophyll were more obviously changed. Malondialdehyde (MDA) is a membrane lipid peroxidation in the final product, it can inhibit cell protective enzyme activity and lower antioxidant content, thereby increasing membrane lipid peroxidation, It reflects the degree of plant injury. MDA analysis showed that the transgenic plants is always lower than that of control content. Chlorophyll of transgenic plants maintain a higher level. So antisense- PLDγgene mainly inhibits membrane lipid peroxidation and thus delays the start of aging.4. A wide variation of transgenic corn plants height. Summer exhibited significantly more dwarf than the control, and the difference at a very significant level. The clustering analysis of plant height in greenhouse, we can see that the height of Zheng58 T0 generation , has a discrete separation, besides of several special height, such as the highest 177cm,168cm, most dwarf 34cm. In the distance of 5.60, the entire group can be divided into four major groups. The separation of the types about L9801 T0 generation height are richer than Zheng58's, and in the distance of 7.40, the height can be divided into six groups. T1 is T0 generation greenhouse harvest species into a single spike of strains, various strains of individuals within the basic uniform. Notable analysis showed that the plant height and ear have the very high significant difference. As soon as T1 was concerned, the height and other agronomic traits are also differences, the similar results with T0 height analysis. T2 height maintaines T1 height variations.5. Transgenic lines permeability analysis. The electrical conductivity values of Zheng58 T0 generation plants between 1.787~3.611 showe discontinuous changes .If opened from the shortest distance of 0.12, the data can be divided into 11 categories, except for two relatively concentrated, others single or for a small decorated. The conductivity of T0 generation L9801 changes in the scope of 1.143~2.272, divided into five categories in the shortest distance of 0.09, 0.12 distance divided into four classes, and differences among classes are significant. Leaves are right from the same location in T1 plants, handled with 1%NaCL in two days, four days, six days and then measured the conductivity. With the days increasing to deal with ,the conductivity is increased, but the transgenic lines were always lower than the corresponding control. It showed that antisense-PLDγgenes can increase the effects of salt or drought tolerance.6. The wide variation of agronomic traits in transgenic lines. The agronomic traits in transgenic lines with receptor have significant difference, such as plant height, growth rate, variations of Picayune. Transgenic plant named Z4.6 matures earlier than the receptor about 6-10 days, This is extremely beneficial to improve receptor shortcomings of mature late. It is observed more variability of maize, involving various characteristics of plant and seeds, including stem color, grain color, grain size and the tolerance to herbicide.7. The biological software analysis of the transgenic fragment. It showed that it is difficult of the transfering antisense-PLDγgene to be translated into peptide in the future generations, but it will be found many small RNAs. The expression of transgenic fragment may be due to insertion points arising to the rearrangement of DNA or gene silencing. Maybe the transcription of the fragment in nuclear is cut into many small RNAs, and roles in the form of RNA interference in the relevant gene. The fragment may be transcribed into mRNA ,affects transcription of target gene mRNA by matching the mRNA.
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