西瓜遗传转化体系的优化

Optimization of the genetic transformation system in watermelon

摘要

以8种基因型西瓜为试验材料，通过统计不同基因型材料的发芽率、不定芽诱导率，筛选出再生能力较强的基因型，并以其为材料对种子播种后培养时间、外植体造伤方式、菌液浓度、共培养时间及侵染方式等进行优化筛选，旨在建立高效的西瓜遗传转化体系。结果表明，在播种2 d时，YL、CYY298、CYY171及CYY137之间的种子发芽率差异不显著且均显著高于其他材料，但在不同浓度GA₃诱导下，YL外植体的不定芽诱导率均最高；以YL材料为基础进行体系优化，发现播种2 d后的种子状态最适于侵染；刀片划伤后的外植体荧光亮度最强，荧光率最高达74.9%；菌液浓度OD₆₀₀=0.8，共培养时间为3 d时，外植体分化状态最佳且荧光率最高为70.3%；真空负压方式可以显著提高侵染效率，外植体荧光率可达71.8%，愈伤组织诱导率高达60.2%；分化培养基中添加0.5 mg·L^-1的GA₃后不定芽的诱导率最高为74%；生根培养基中添加0.5 mg·L^-1的IBA时生根率最高，平均生根数最多。

Abstract

In this study, eight watermelon genotypes were used as experimental materials, the germination rates and adventitious buds induction rates of different genotypes were counted and analyzed to screen out the optimal genotype with strong regeneration ability. The efficient watermelon genetic transformation system was optimized and screened through different sowing times, explant injury methods, bacterial liquid concentration, co-culture time, infection methods and other factors. The results showed that the seed germination rates of YL, CYY298, CYY171 and CYY137 had no difference and were significantly higher than that of other materials at 2 days of seeding, but the germination induction rate of YL was the highest at different concentrations of GA₃. The system was optimized based on YL material, and it was found that the seed germination state was the best and most suitable for infection after 2 days of sowing. The fluorescence brightness of the explants after scratching the blade was the strongest, and the fluorescence rate was up to 74.9%. When the concentration of bacterial solution was OD₆₀₀=0.8 and the co-culture time was 3 days, the explants were in the best differentiation state and the fluorescence rate was the highest at 70.3%. The infection efficiency can be significantly improved by negative pressure infection, and the fluorescence rate of explants can reach 71.8%, and the callus induction rate can reach 60.2%. The induction of adventitious shoots was up to 74% after the addition of 0.5 mg·L^-1 GA₃ to the differentiation medium. The highest rooting rate and the highest average number of roots were observed when 0.5 mg·L^-1 of IBA was added to the rooting medium.

关键词

西瓜 / 遗传转化 / 转化效率 / 体系优化

Key words

Watermelon / Genetic transformation / Transformation efficiency / System optimization

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基金资助

河南省科技研发计划联合基金(222103810009)

河南省重大科技专项(221100110400)

河南省农业良种联合攻关项目(2022010503)