| The experiments have been conducted in the induction, multiplication and differentiation of anther cullus with three eggplant varieties ( Yuyan No. 1 ,Sanyueqie,Pingzaoqiu No. 1).And then a few haploid plants were obtained. In addition, further research on the triggering and growth of the microspores and the differentiation of the anther callus has been done at the cytohistology, physiology and biochemistry levels.The main results are as follows:1 Selection and pretreatment of materialAlabastrums of different development phases were dyed with I-KI, pressed with cover glass and examined with microscope so that the anthers were selected for tissue culture whose pollens were developing at mid or late uninucleate stage.The relativity between exterior shape of alabastrums and growth phases of pollens would probably differed considering different varieties and environment. In a general way, the phase with about seven cilimetre length anther was suitable when calyx was going to dehisce but not open and corolla with aqua was included in sepal.Then the majority of alabastrums picked were enwrapped with waterish pledget and pretreated with5~6℃ in refrigeratory, and the others inoculated were cultured at the room temperature (CK) or treated at the high temperature.2 Good disinfection system of eggplant alabastrumsDisinfection of alabastrums was fundmental to anther culture because eggplant alabastrums have a lot of glosses on their surface.The results indicated that the disinfection treatment of NaClO was noneffective, and the only HgCl2 was also inapparent,but pretreating30s with 70% alcohol before disinfecting 10~12min with 0.1% HgCl2 supplemented with a bolb of tween-20 was efficient and high qualitative.3 Induction of anther callusThe best condition of induction was selected about the best combination of hormone and organic compound, the optimum time of pretreatment by low temperature, the optimal time of treatment by high temperature and the suitable concentration of sucrose after the anthers were put into the induction media, the results indicated that there were many influence factors to the induction of anther callus.The optimal conbination of hormone and organic compound was 2,4-D1 .0mg/L + KT 0.5mg/L + LH1g/L + Gln500mg/L.The best concentration of sucrose was 30~40g/L.The most suitable time of low temperature pretreatment was 48-72h at 5~6℃, and the optimum time of high temperature treatment was 48~60h at 33℃.However, the influence of low temperature pretreatment couldn't interact with that of high temperature treatment to increasing the induction rate. Under the optimal condition including forementioned factors, the callus induction rate of Yuyan No.1.Sanyueqie and Pingzaoqiu No.1 were about 12.00%,7.00%,25.00% respectively.4 Differentiation of anther callus and rhizogenesis of adventitious budMorphogenetic callus by inducing and proliferating culture was transferred into different differentiation media to induce adventitious buds. The results indicated that the optimum medium of differentiation was MS + ZT1.0mg/L +NAA0.001 -0.005mg/L+ GA30.5mg/L+ LH1g/L. Under this condition, the differentiation rate of Yuyan No.1, Yanyueqie and Pingzaoqiu No.1 attained 33.33%, 24.00% and 68.89% respectively. Then the adventitious buds were transferred into (he rhizogenesis medium of 1/2MS supplemented with 0~0.005mg/LNAA and 20g/L sucrose,and rooted well. When aftergrowthes were transplanted the livability was more than 90%.5 Identification of haploid plant and diversification of chromosome multipleThe chromosome multiple of root tip of aftergrowth was identified with the way of ridding cell wall and putting low osmosis.The results showed that the percentage of haploids, diploids, triploids, tetraploids and aneuploids-mixoploids were 16.84%, 54.74%,4.21%, 9.47% and 14.74% respectively at 95 aftergrowthes by anther culture. But the proportion of chromosome multiple was different at the anther aftergrowthes of three genotypes.6 Observation of microspore triggering and growthIn the cou... |
PDF Full Text Request