| In this paper the pollens of the species of Fraxinus Linn were treated with high voltage static, which promoted the cross-compatibility by enhancing the vitality of their pollen. In order to study the molecule mechanism of it, both of the control and pollens dealt with high voltage static field were taken for RNA extract.The DNA fragments of differential expressed genes were gotten by DDRT-PCR and were confirmed by Reverse Northern dot blotting, and then they were sequenced and analyzed. The main results were as follows:1, The suitable culture medium for the pollens germination of Fraxinus Linn is : sugar 10 g.L-1+agar 1 g·L-1+urea 0.1 g·L-1+boric acid 0.5 g·L-1+nitre 0.1 g·L-1+bitter salt 0.2 g·L-1 +calcium nitrate 0.1 g·L-1+zinc salfate 0.1 mg·L-1+sodium molybdate 20 mg·L-1+NAA10 mg·L-1+6-BA 1mg·L-1,pH 6.5.2,The differences of pollens vitality between individual plants were significant, which up to 30.9 percent at the most among the trees experimented in this paper.3,To keep higher vitality ,pollens should be sealed after dry and stored below 4℃in refrigerator.4,The germination rate of pollen of Fraxinus mandshurica treated with the proper high voltage static field was 19.7 percent higher than that of control.5,The pollens treated with proper high voltage static field could promote hybrid between the species of Fraxinus Linn and get a higher fructification rate, 13.5 percents higher than control at the most.6,The hybrid seeds of Fraxinus sogdiana Bunge, Fraxinus velutina Torr. and Fraxinus rhynchophylla obtained by pollen processed by proper high voltage static shared higher data in seed length, seed width and kilo granule weight than control, and the higher values of these were0.25centimeters(9.40percents),0.06centimeters(9.84percents)and13.71gram (25.9percents) at the most.7,F1 seedlings from pollen treated by high voltage static were notably different from control(not treatment)in seedling height, ground stem width, photosynthesis rate and chlorophyll content.8,Improved CTAB method is effective in extracting the RNA in the pollen of Fraxinus mandshurica.9,Genes in the pollen of Fraxinus mandshurica expressed differential after processed by high voltage static field. Identified by reverse northern dot blotting, 6 differential expressed gene fragments were gotten. By sequencing and analyzing, we found that the fragments have great identity with the gene for cyclic nucleotide-gated cation channel in Arabidopsis thaliana, with the gene for calmodulin binding / cyclic nucleotide binding / ion channel in Arabidopsis thaliana, with the gene for chloroplast Cu-Zn superoxide dismutase in tomato,with the gene for putative cyclic nucleotide-regulated ion channel protein in Arabidopsis thaIiana, with the gene for auxin-induced proteinase inhibitor (ARPI) gene in Lycopersicon esculentum, and with the cytochrome P450 monooxygenase CYP76E1 (CYP76E1) gene in Medicago truncatula respectively.There nucleotide homologous were 98percents, 95percents, 96percents, 99percents, 93percents and 93percents.
|
PDF Full Text Request