| Studies on in vitro conservation of several fruit germ plasm by restricting growth methodsIt's very important to conserve fruit germplasm, and there were many kinds of methods of conservation. In vitro conservations including restricting growth method built on plant tissue culture. Some stem apex culture system of apple (Luo6, lingdangguo.etc) varieties were established in this study, which provided materials for the study of effect of conservation by adding different medicine in MS culture medium and by changing culture condition. The genetic aberrance of the materials before and after the conservation was studied by RAPD analysis methods. After having been conserved for one year, the micropropagation and producing of Luo6 were studied. The main results were as follow:1. Stem apex culture system of Luo6 was established, which provided material for the studies on conservation and genetic variationA lot of stem apex culture mediums of micropropagating about Luo6 were established by many experiments. The best differentiation medium of Luo6 was MS+BAo 45+IBAo i , in which the ratio of differentiation was highest and the leaves of the stem apex culture plantlet of Luo6 were green. During establishment of the stem apex culture of Luo6, brown change was also studied and many methods of conquering it were obtained.2. Methods of restricting growth of fruit stem apex culture plantlet was improved and perfectedDecreasing the volume of the air in container, adding 2% sorbitol in MS culture medium, at low temperature of 5'C, reducing the time of illumination to 6 hours every day or giving 500 Lux of illumination , increasing agar concentration to 11 g/L or adding active carbon concentration to lOg/L, some apple and strawberry varieties studied were conserved for 18 months till now. The growth speed of fruit stem apex culture plantlet by restricting methods was distinctly restricted than control, and the plantlet after conservation can normally grow and differentiate after transferring to normal culture medium when given normal culture condition. In the present paper, the glass-like of stem apex culture plantlet during conservation was also studied and solved.3. Genetic varieties of stem apex culture plantlet conserved for 1 year were studied by molecular marker of RAPDUsing molecular marker of RAPD with 14 random primes, genetic varieties of different varieties of apple and strawberry before and after conservation by different restricting growth methods were studied. 224 strips were obtained which were in order and uniform, and there were no different strips found. All these showed that the genetic type before and after conservation were consistent and there were no variation found.434. Technique program of system conservation of in vitro fruit germplasm was establishedBefore the conservation of fruit stem apex culture plantlet, with suitable medium which contained 7g/L agar ,30g/L sucrose and other suitable medicine, materials to be conserved were first cultured in normal conditions such as 2000 Lux illumination every day, 25 "C of temperature, et. After the stem apex culture plantlet had grown for two weeks and more haleness, using small glass bottle of penicillin bottle as culture bottle, the stem apex culture plantlet was first cut and inoculated in normal MS medium adding 2% sorbitol, and then it was placed in illuminating incubator and was cultured at 5癈 with 6-hour-long 2000 Lux illumination in order to be conserved. The status of growth of stem apex plantlet conserved should be detected every two weeks.5. System of micropropagation and rooting about Luo6 conserved was studiedThe best elongation medium of Luo6 conserved was : MS+KT2.o-t-NAAo.i,in which the elongation of Luo6 was distinctive and its leaf was plump. The best shooting medium of Luo6 conserved inside culture bottle was: l/2MS+IBA0.2,in which medium Luo6 took root after 9 days and there were more shoot in Luo6. The growth of stem apex culture plantlet of Luo 6 after rooting was haleness, which was propitious to trans...
|
PDF Full Text Request