兔眼蓝莓组培体系建立及多倍体诱导研究

Establishment of Vaccinium ashei Tissue Culture System and Polyploidy Induction

蓝莓具有较高的营养价值和经济价值,但其繁殖速度慢、果实较小、采摘成本高,优化现有的蓝莓品种具有良好的发展前景。本文以兔眼蓝莓灿烂幼嫩茎段为试验材料开展组织培养研究,采用不同的培养基配比激素以及设置不同浓度用以研究出最适合兔眼蓝莓灿烂生长的、稳定的、可重复性强的培养基配方,建立叶片再生体系,并采用秋水仙素培养基添加法对其进行诱导。在无菌条件下切取由叶片再生形成的长势较好苗的带叶茎尖(长约0.5 cm),接种到秋水仙素浓度为10、20、30、40、50、60 mg/L的6种培养基中。结果表明,最适兔眼蓝莓灿烂腋芽启动培养基是配方WPM+0.005 mg/L NAA+2.0 mg/L ZT+20 g/L蔗糖+6 g/L琼脂,诱导效果最佳,发芽率高达93%以上;取离体叶片再生芽苗的茎尖进行共培养诱导多倍体,50 mg/L秋水仙素处理50 d和60 mg/L秋水仙素处理30 d或40 d时,诱导植株变异的效果最佳,变异率均达30%以上。

Blueberry has high nutritional value and economic value,but its propagation speed is slow,the fruit is small,the harvesting cost is high,so the optimization of existing blueberry varieties has a good development prospect.In this study,the young stem segment of Vaccinium ashei′Brilliant′was used as the experimental material to carry out tissue culture research.Different ratios and concentrations of hormones were used to develop the most suitable component for the stable and reproducible culture medium of V.ashei′Brilliant′.The leaf regeneration system and polyploidy induction system were established,and it was induced by colchicine medium addition method.Under sterile conditions,the leaf tips of the growing seedlings(about 0.5 cm long)formed by leaf regeneration were inoculated into 6 mediums containing colchicine concentration of 10,20,30,40,50,60 mg/L.The results showed that the optimal axillary bud initiation medium for V.ashei′Brilliant′was WPM+0.005 mg/L NAA+2.0 mg/L ZT+20 g/L sucrose+6 g/L agar,the inducing effect was the best,and the germination rate was over 93%.The leaf tips of the growing seedlings formed by leaf regeneration in vitro was co-cultured to induce polyploidy,when treated with 50 mg/L colchicine for 50 d and 60 mg/L colchicine for 30 d or 40 d,the plant variation effect was the best,the variation rate was over 30%.