Study On Tissue Culture Of Prunus Humilis

To explor the efficient way in vitro culture, the stems of Prunus Humilis wereutilized to induce callus which could induce cluster buds to multiply. In addition, the stemtips were directly induced into adventitious buds to multiply. Experiment was designedprimarily by single factor and orthogonal design. Through variance analysis and multiplecomparisons, we found out a series of technical parameter combination for the tissueculture of Prunus Humilis, which provided the scientifical and technical support for itsfactorization production. The main results gere as follows:1. The best type of explants in vitro was the mid-stems with axillary buds, whichshowed lower contaminated rate and higher differentitation rate.2. The optimum sampling period of Prunus Humilis was in April. In the growingstage, the tissue culture showed high germinating rate, low contaminated rate and theyoung plants grew quickly. At the same time, effects of sterilization time on contaminatedrate and embryonic rate of Prunus Humilis was different. The explants sterilized for 8minutes was the best treatment.3. The proper hormones for inducing callus were NAA and BA. The treatment of1/2MS+BA2.5 mg/L+NAA0.8 mg/L was the most suitable hormone matching of callusinducement. The callus of this treatment was green and inseparable.4. The treatment of MS+BA0.5 mg/L+NAA 0.5 mg/L was the most suitable stem tipculture medium for the differentiation of adventitious buds with thick stems, higherdifferentiation rate, forming shoot at one time.5. MS culture medium was the optimum medium for multiple.6. NAA and BA were the suitable auxins and cytokines respectively in vitro cultureof Prunus Humilis, and the best multiplication efficiency was obtained in the treatment ofNAA0.06 mg/L+BA1.0 mg/L.In addition, bunch buds of Prunus Humilis grew slowly, sothe multiplication cycle should be prolonged to 45 days.7. Prunus Humilis has wide accommodation of pH 5.8～6.2, but pH 5.8 was the besttreatment.8. Single form of nitrogen could not satisfied the multiplication of PrunusHumilis. On the basis of MS, high-concentration NO₃^- could promote multiplication of Prunus Humilis. Different ratios of NH₄⁺ and NO₃^- in the medium had differentresults, NH₄⁺: NO₃^-1:2 was the best ratio.9. There was no signifficant difference in multiplication efficiency between the twotreatments of cane sugar and sand sugar as carbon source, so the cost of tissue culturecould be reduced.10. The tube young plants of Prunus Humilis root slowly, and the seed with 2.0 cmheight, two leaves and better growth rooted well. The proper auxin for rooting was IBA. Inaddition, 1/2MS+IBA0.05 mg/L was the best treatment. The roots of this treatmentshowed proper length and thickness, more side roots and higher rooting rate.11. The suitable acclimatization time was 7～10 days. The well-penetrated substanceadapted to the plant transplantation in the initial stage and pure vermiculite was the mostsuitable transplantation substance.