以40个不同基因型万寿菊(Tagetes erecta)叶片为外植体, 在相同条件下诱导不定芽分化, 获得最佳再生基因型; 然后分析不同激素组合、外植体切口方式、固化剂及蔗糖对万寿菊再生和玻璃化影响; 最后对不同类型的伸长培养基进行探索。结果表明, 最佳再生基因型为里程碑·黄色; 最佳再生培养基为MS+0.2 mg·L^‒1 TDZ+0.5 mg·L^‒1 IBA+8 g·L^‒1琼脂+40 g·L^‒1蔗糖, 再生率达70%, 玻璃化率降低至16%; 最适再生的小叶部位为全小叶; 最适伸长培养基为MS+8 g·L^‒1琼脂+30 g·L^‒1蔗糖, 伸长率达91.3%。该研究建立了高效稳定的万寿菊再生体系, 解决了万寿菊再生过程中严重的玻璃化问题, 可为万寿菊的遗传改良和基因功能研究奠定基础。

In this study, marigold (Tagetes erecta) leaves of 40 different genotypes were used as explants to induce adventitious buds formation to select the best regenerating genotype. In addition, main factors impacting the selected genotype’s regeneration and vitrification were analyzed, including hormone combinations, the way of explant incision, curing agent and sucrose, etc. Finally, different types of elongation medium were explored. The best regenerated genotype was marigold Milestone·Yellow. The best regeneration medium was MS media supplied with 0.2 mg·L^‒1 TDZ, 0.5 mg·L^‒1 IBA, 8 g·L^‒1 agar, and 40 g·L^‒1 sucrose, in which the regeneration rate was 70% and the vitrification rate was reduced to 16%. Whole leaflet was the most suitable explant among different leaflet segments; the best performing elongation medium was MS media supplied with 8 g·L^‒1 agar and 30 g·L^‒1 sucrose, and the elongation rate was 91.3%. This study establishes an efficient and stable regeneration system of marigold, and solved the serious vitrification problem commonly found during regeneration. It laid a foundation for marigold genetic improvement and gene function research.