Studies On In Vitro Culture Of Chinese Jujube And Sour Jujube

Chinese jujube and sour jujube originated in China belongs to Ziziphus genus, Rhamnaceae family, and is an important resource in our country. This study revealed effects of optimal stage, pretreatment, concentrations of sucrose and plant growth regulator on anther culture in wild jujube and'Zanhuangdazao','Pingguozao','Junzao', and obtained plant regeneration of anther. Callus were induced from leaves, stem and anther of wild jujube, and obtained the best combination of plant growth regulator; The research isolated protoplast from pollen and suspension cell lines of wild jujube, and found the best concentration of enzyme, enzemolysis time and concentration of mannital. The study was the foundation to early times of ploidy breeding. Experiment was carried out with leaves in vitro of sour jujube for studying the effects of adventitious shoot regeneration, elongation of adventitious shoot regeneration, proliferation culture of plant regeneration, rooting culture of plant regeneration. The highly effective plant regeneration system of sour jujube leaf was established. Through anther culture, proliferation culture and the highly effective plant regeneration system of leaf was beneficial to germplasm enhancement and new variety breeding. Lay a foundation for germplasm resources in vitro conservation, mutation breeding, clones from single bud, transgenic acceptor system and rapid propagation in vitro clonal stock. It had theoretical value and practical meaning.The main results were as followed:1 Obtained plant regeneration on anther of'Zanhuangdazao'and'Pingguozao'(1) The difference times of low temperature treatment significantly influenced anther of'Zanhuangdazao'and'Pingguozao'. Low temperature treated'Zanhuangdazao'anther for 3 days, the induction rate of callus was the biggest and it was 79.27%. Low temperature treatment was helpful to callus differentiation and plant regeneration. Low temperature treated'Pingguozao'anther for 3 days, the induction rate of callus was the biggest and it was 87.71%. Low temperature treatment was helpful to callus differentiation and plant regeneration.(2) When concentrations of sucrose were 6% and 3%, which may help induce anther callus on'Zanhuangdazao'. But when concentrations of sucrose were 6%, the browning rate of callus was higher. The sucrose concentration of 3% induced helpfully anther callus. When concentrations of sucrose was 3%, which may help induce anther callus on'Pingguozao'.(3) Callus could be efficiently induced from anther in the medium MS+TDZ+NAA+30 g/L sucrose+6 g/L agar. TDZ 0.5 mg/L and NAA 1.5 mg/L were suitable for callus induction of anther of'Zanhuangdazao'and'Pingguozao'. The induction rate of callus of'Zanhuangdazao'reached 87.71%, the induction rate of callus of'Zanhuangdazao'reached 92.01%.(4) Callus could be efficiently differentiated from anther in the medium MS+TDZ+NAA+30 g/L sucrose+6 g/L agar. 6-BA 1.5 mg/L and IBA 0.1 mg/L were suitable for callus differentiation of anther of'Zanhuangdazao'and'Pingguozao'. The differentiation rate of callus reached respectively 76.60% and 83.48%. The regeneration coefficient reached 6.05 and 12.36 respectively. This study obtained plant regeneration of anther, the anther of'Zanhuangdazao'was 21 tissue culture seedlings and the anther of'Pingguozao'anther was 45 tissue culture seedlings.2 Proliferation culture of'Zanhuangdazao'and'Pingguozao'(1) The best proliferation medium for the plant regeneration of'Zanhuangdazao'was MS+6-BA 1.5mg/L+IBA 0.1mg/L. The elongation was 2.57 cm and the multiplication coefficient was 5.14。(2) The best proliferation medium for the plant regeneration of'Pingguozao'was MS+6-BA 1.0mg/L+IBA 0.1mg/L. The elongation was 3.82 cm and the multiplication coefficient was 4.77。3 Gaining the high efficient plant regeneration in sour jujube in vitro leaves(1) It was found by study that dark culture was necessary for leaf regeneration of sour jujube. The leaves inoculated on the regeneration medium, a period of darkness for 3 weeks was necessary to increasing the regeneration rate. The number of adventitious shoots from per leaves was significantly enhanced.(2) The different plant growth regulator had different effects on inducement of regeneration from leaves of sour jujube. The cytokinin, both of 6-BA and TDZ could induce regeneration of leaves of sour jujube. 6-BA induced leaves regeneration, there was little callus, and the adventitious shoots regenerated from the leaves directly. The adventitious shoots grew shedding shoot of jujube. TDZ induced leaves regeneration, there was much callus and the adventitious shoots regenerated from the callus. The adventitious shoots grew vegetative shoot of jujube.The optimum cytokinin was TDZ and its concentration was 0.5mg/L. Lower concentration of auxin could induce regeneration of leaves of sour jujube, the optimum auxin was IBA 0.1mg/L.(3) The best medium of the adventitious shoot regeneration of sour jujube was MS+ TDZ 0.5mg/L+IBA 0.1mg/L. The adventitious shoots regeneration rate was 93.57% and the number of adventitious shoots from per leaves was 4.23. (4) The best elongation medium of the adventitious shoot regeneration of sour jujube was MS+6-BA 0.5mg/L+IBA 0.1mg/L+GA3 0.5mg/L. GA3 could significantly increase the elongation of adventitious shoots.(5) The best proliferation medium for the plant regeneration of sour jujube was MS+6-BA 0.5mg/L+IBA 0.2mg/L. The elongation was 3.07 cm and the multiplication coefficient was 6.13。(6) The best rooting medium for the plant regeneration of sour jujube was 1/2 MS +IBA 1.5mg/L. The rooting percentage was 67.92% and the average roots number was 3.21.