Tissue Culture And Establishment Of Rapid Propagation System Of Blueberry ’Tiffany Blue’

The industrialized seedling cultivation of blueberry(Semen Trigonellae)is mainly based on seedling propagation,while the method of seedling propagation is difficult to achieve large-scale production of high-quality seedlings,and the tissue culture technology has the advantages of high propagation coefficient and easy seedling formation.In this study,the young branches of Rabbiteye blueberry were used as test materials to study the effects of sterilization agents,culture media,plant growth regulators and seedling refining media on the sterilization of explants,leaf regeneration,subculture proliferation,rooting and seedling refining transplantation,in order to provide theoretical basis for blueberry factory seedling cultivation.The main test results are as follows:According to the situation of the blueberry industry in Changsha area,’Tifu Blue’ was selected as the experimental material.The optimal sterilization method for Tiffany Blue’explants was to sterilize them with 75%alcohol for 30 scconds,then sterilize them in 10%NaClO for 15 minutes,and finally rinse them with sterile water for 3-5 times.The axillary bud germination rate reached 68.3%’The optimum medium for primary culture of Tiffany Blue ’was MWPM·MS+1.0 mg/L ZT+0.2 mg/L NAA.The optimum medium for leaf callus induction was 1/2WPM+2.0 mg/L ZT+0.5 mg/L 2,4-D+30 g/L sucrose,the callus induction rate was 91.3%,and the browning rate was 8.5%.In the process of callus subculture,40 mg/L PVP added to the medium can effectively inhibit the browning of callus.The optimal combination of adventitious bud differentiation medium is 1/2 WPM+2.0 mg/L ZT+2.0 mg/L TDZ+ 0.5 mg/L NAA+ 30 g/L sucrose.After dark culture for 2 days,the optimal effect of adventitious bud induction is 75.2%,and the adventitious bud induction coefficient is 8.4.The optimal proliferation medium was 1/2WPM+0.1 mg/L NAA+1.5 mg/L ZT+20 g/L sucrose,the proliferation coefficient and average plant height were 5.5 and 5.2 cm,respectively,and the proliferation seedlings were relatively robust;After subculture for 5 times,the effect of proliferation culture was better;Using 2.0 mg/L AgNO3 and 15 mg/L PVP as anti browning agents can effectively inhibit browning of tissue cultured seedlings in the process of adventitious bud proliferation and culture’ The optimum culture medium for Tiffany Blue was 1/2 WPM+0.5 mg/L IBA+200 mg/L AC,and the rooting rate was 88.4%and the average rooting number was 6.83 after 7 days of dark culture;Adding gibberellin in root formation stage could release dormancy;The rooting effect of out of bottle rooting method is not ideal,the main reason is that the pollution rate is too high;The most suitable transplanting substrate is mossy soil;The best transplanting season is autumn.Soaking in 1.5 mg/L IBA solution for 2 hours before transplanting can effectively improve the transplanting effect.After transplanting,new adventitious roots will emerge from the roots,with a regeneration rate of 65.7%and an average root length of 7.8 cm.In this study,stem segments with buds of Tiffany Blue’ were used as explants,and sterile seedlings of Tiffany Blue were successfully obtained through disinfection treatment.The leaves of sterile seedlings were used as basic materials to induce callus and adventitious buds through leaves,and a regeneration system of leaves was established.The adventitious buds induced by callus tissue were propagated and cultured to obtain a large number of vigorous tissue culture seedlings,The tissue culture system of Tiffany Blue has been successfully established,which provides a scientific basis for the subsequent genetic transformation and industrialized seedling cultivation.