苦瓜离体再生体系建立的研究

Study on the Establishment of in vitro Regeneration System in Momordica charontia L.

[目的]研究影响苦瓜离体再生体系建立的条件,为苦瓜遗传转化体系的建立奠定基础。[方法]以碧绿二号、丰绿、共和、早绿和大顶苦瓜品种无菌实生苗为试材,探讨了不同前处理、不同激素组合及培养条件对苦瓜离体形态发生的影响。[结果]播种前对苦瓜种子进行预处理能明显提高发芽率和发芽势,苦瓜愈伤组织的诱导率比较高,不受基因型和外植体种类的限制;选用上胚轴为外植体的愈伤组织发生效果较其他外植体好;12种外源激素组合中,较强分裂素类物质ZT与激动素KT组合,对愈伤组织的分化有促进作用,以MS+ZT4 mg/L+KT1 mg/L分化培养基为最佳;苦瓜愈伤组织的再分化用琼脂固化的MS培养基比液态MS培养基效果好;pH值过低或过高都不适宜愈伤组织分化,以5.8为最适。[结论]该研究为苦瓜离体再生体系的建立提供了理论基础。

[Objective] The research aimed to study the conditions that affected the establishment of in vitro regeneration system in Momordica charontia L. and lay the foundation for the establishment of the genetic transformation system in M. charontia. [Method] With the aseptic seedlings of M. charontia varieties including Bilv 2, Fenglv, Gonghe, Zaolv and Dading as tested materials, the effects of different pretreatment, different hormone combinations and culture conditions on the morphogenesis in vitro in M. charontia were discussed. [Result] Pretreating the seeds of M. charontia before sowing could obviously increase the germination rate and the germination potential. The callus induction rate in M. charontia was higher and it was not restricted by genotype and explants types. The effect of inducing callus by selecting epicotyl as explant was better than other explants. Among 12 kinds of exogenous hormone combinations, the combination of strong mitogen ZT and kinetin （KT） had the promotion on the callus differentiation, with the differentiation medium of MS＋ZT 4 mg/L＋ KT 1 mg/L best. The callus redifferentiation of M. charontia on agar-solidified MS medium had better effects than that on liquid MS medium. Too low or too high pH value was not suitable for the callus differentiation, with the optimum pH value of 5.8. [Conclusion] The research provided the theoretical basis for the establishment of in vitro regeneration system in M. charontia.