In Vitro Anther Culture Of Five Chinese Jujube Cultivars Including ’Yueguang’

In the study, effects of some factors including genotypes, basic medium, hormones,carbon source and colchicines on anther culture of Chinese jujube were investegated usingfive cultivars, i.e.,‘Yueguang’,‘Dongzao’,‘Changhongzao’,‘Pingguozao’ and‘Jinsixiaozao’. Anther plants of ‘Changhong’ and ‘Pingguozao’ was firstly obtained. Anoptimized regeneration system for anther plants of ‘Yueguang’,‘Dongzao’ and‘Jinsixiaozao’ was established. A pollen plant (Diploid) was obtained in culture of‘Yueguang’ anther treated by colchicines, which contained2n gametes. An propagationsystem for anther plants of ‘Yueguang’ was established. In addition, the recoverytechniques of vitrification plants was systematically studied. The results were as follows:1Callus induction from anther of Chinese jujubeCallus formation from anther in Chinese jujube was influenced significantly bygenotype, plant growth regulators, basic medium, carbon source.20g/L maltose promotedto induce callus for Chinese jujube. The optimal medium for callus induction was MSmedium supplemented with1.0mg/L2,4-D,2.0mg/L6-BA,5.5g/L agar and20g/Lmaltose.‘Changhongzao’ callus rate was100%,‘Yueguang’,‘Yueguang’ jujube appliedcolchicines,‘Pingguozao’ and ‘Dongzao’ callus rate was more than80%, browning rate ofcallus was less than22%. The optimal medium for ‘Jinsixiaozao’ callus induction was1/2MS medium supplemented with0.5mg/L NAA,2.0mg/L6-BA,5.5g/L agar and20g/Lmaltose, callus rate was68.2%, browning rate of callus was13.0%.Colchicines promoted to induce callus for ‘Yueguang’. The effect of mediumsscreened was stable by anther callus induction of ‘Yueguang’ and ‘Yueguang’ jujubeapplied colchicines in different years.2Proliferation and differentiation of anther callus in Chinese jujubeProliferation and differentiation of callus from Chinese jujube anther was significantlyaffected by genotype and plant growth regulators. The optimal medium for plants inductionwas MS medium supplemented with0.3~0.5mg/L NAA,1.0~1.5mg/L6-BA,5.5g/L agarand20g/L Sucrose. In five varieties of Chinese jujube,‘Yueguang’ had the maximumbudding rate (86.9%),‘Jinsixiaozao’ budding rate was only20.4%. The sprouting rate was not obviously correlated with the number of plants per callus obtained, but browning ratesignificantly affect the shoot regeneration rate of callus.High concentrations of cytokinin was in favor of callus proliferation, but increased thevitrification degree of plants.When medium supplemented with AgNO3, number of plants from callus wasincreased, but plants regenerate time was extended, callus budding rate was decreased, thewhole plant were small and grew slow, callus proliferation was slow with browningseriously. In short, bud differentiation medium unfit added AgNO3.Colchicine was conducive to plant regeneration for Chinese jujube anther culture,could reduce the callus browning rate and improve the shoot regeneration rate of callus.3Vitrification plants recovery0.4mg/L IBA+1.0mg/L6-BA promoted to subculture vitrification anther plants ofChinese jujube, Average increment per plant was1.93cm. The optimal medium forvitrification plants recovery was MS medium supplemented with0.4mg/L IBA,1.0mg/L6-BA,2g/L PVA,5.5g/L agar and7.5g/L maltose,15g/L Sucrose. The optimal sealmaterial was sealing film. The suitable temperature was26℃. Average plant heightbefore processing was above1.5cm promoting vitrification plants recovery.4Identification the origin of anther plantlets125anther plants of ‘Yueguang’ and210anther plants of ‘Yueguang’ jujube appliedcolchicines were analyzed with seven SSR primer pairs to determine their source of origin,we found the phenomenon which a lack of electrophoretic bands appeared andelectrophoretic bands difference in position for one pollen plant. Plants ploidy level weredeterminated by flow cytometry, measured30strains were diploid. The pollen plant whichwas a diploid plant was obtained from anther culture of ‘Yueguang’ jujube appliedcolchicines.