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中国农学通报 ›› 2015, Vol. 31 ›› Issue (10): 80-86.doi: 10.11924/j.issn.1000-6850.casb15010107

所属专题: 园艺

• 林学 园艺 园林 • 上一篇    下一篇

黄瓜离体器官再生体系的优化

钟辉丽,李玉红,陈雪琳,由曜华   

  1. (西北农林科技大学园艺学院,陕西杨陵 712100)
  • 收稿日期:2015-01-15 修回日期:2015-03-05 接受日期:2015-01-26 出版日期:2015-05-05 发布日期:2015-05-05
  • 通讯作者: 李玉红
  • 基金资助:
    国家自然基金项目“黄瓜矮生基因(cp)的精细定位、克隆及功能分析”(311719553)。

Optimization of Regeneration System in Cucumis sativus L.

Zhong Huili, Li Yuhong, Chen Xuelin, You Yaohua   

  1. (College of Horticulture, Northwest A&F Univorsity, Yangling Shaanxi 712100)
  • Received:2015-01-15 Revised:2015-03-05 Accepted:2015-01-26 Online:2015-05-05 Published:2015-05-05

摘要: 优化通过器官直接再生方式的黄瓜离体再生体系,为遗传转化奠定基础。以‘长春密刺’黄瓜的子叶节为外植体,探讨在黄瓜再生过程中,适宜的无菌苗获得培养基、适宜的外植体类型、无菌苗的适宜苗态、不定芽诱导培养基和芽伸长培养基中适宜的激素组合与比例。结果表明,最适的无菌苗获得培养基为1/2MS+30 g/L蔗糖+7 g/L琼脂(pH=5.8);不同外植体的再生率为子叶节>下胚轴>子叶;子叶完全出壳但未展平的无菌苗比其他苗态的再生率高,子叶展平后,再生率迅速下降;当6-BA(6-苄基氨基嘌呤)和ABA(脱落酸)浓度一定时,最适的AgNO3为2 mg/L;当AgNO3浓度一定时,6-BA和ABA浓度的最佳组合为1.5 mg/L 6-BA+0.5 mg/L ABA;不定芽诱导的最适培养基为MS+1.5 mg/L 6-BA+0.5 mg/L ABA+2 mg/L AgNO3,出芽率为90%,每外植体出芽数为3.5;芽伸长培养基中加入0.10 mg/L 6-BA,能够促进再生芽的伸长。本研究成功优化了黄瓜的再生体系,得到了健壮的黄瓜成株。

关键词: 蝴蝶兰, 蝴蝶兰, 新品种, DUS测试指南

Abstract: The objective of this research was to optimize Cucumis regeneration system, so as to make preparation for genetic transformation. The cucumber cultivar (‘Changchunmici’) cotyledon-nods were used as explants to investigate the effect of seed germination mediums, types of explants, seedling stages, phytohormone combinations and their ratios on the shoot-inducing mediums and the shoot elongation mediums. Results showed that higher efficiency of seed germination was obtained easily when the seeding seeds on 1/2MS sucrose 30 g/L agar 7 g/L, pH 5.8. The results indicated that the regeneration rate from different types of explants showed as cotyledon-nod > hypocotyl > cotyledon. The explant of cotyledons completely separated from husk but not flattened had high shoots regeneration rate than other seedling stages. Regeneration rate would reduce quickly when the cotyledons were flattened. When determining the concentration of 6-BA (6-benzyl amino purine) and ABA (Abscisic acid), the optimum of AgNO3 was 2 mg/L. When determining the concentration of 2 mg/L AgNO3 (2 mg/L), the best concentration combination of BA and ABA was 6-BA 1.5 mg/L ABA 0.5 mg/L. The results showed that MS 6-BA 1.5 mg/L ABA 0.5 mg/L AgNO3 2 mg/L medium was the optimal formula. The highest shoot regeneration frequency reached 90% and 3.5 shoots per explant. 6-BA at 0.5 mg/L markedly increased shoot elongation. The regeneration system of cucumber is successfully optimized and the robust cucumbers are obtained.