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中国农学通报 ›› 2014, Vol. 30 ›› Issue (1): 20-25.doi: 10.11924/j.issn.1000-6850.2013-2136

所属专题: 生物技术 园艺

• 林学 园艺 园林 • 上一篇    下一篇

TDZ对橡胶树花药愈伤组织诱导和体细胞胚发生的影响

李玲,管艳,梁国平,黄凤翔,桂明春,田海   

  • 收稿日期:2013-08-07 修回日期:2013-09-11 出版日期:2014-01-05 发布日期:2014-01-05
  • 基金资助:
    国家科技支撑计划“云南天然橡胶产业提升关键技术集成与应用”(2011BAD30B01);云南省产业重大关键技术开发专项计划“橡胶树优良品种体胚植株培养和籽苗繁殖关键技术集成与开发”(云发改高技[2008]1658号);云南农垦重点科技计划项目“高产抗寒橡胶自根幼态无性系的培育研究”(2006XJ03)。

Effects of TDZ on Callus Induction and Somatic Embryogenesis of Hevea brasiliensis Anther

Li Ling, Guan Yan, Liang Guoping, Huang Fengxiang, Gui Mingchun, Tian Hai   

  • Received:2013-08-07 Revised:2013-09-11 Online:2014-01-05 Published:2014-01-05

摘要: 为了提高橡胶树优良品种的愈伤组织诱导和体细胞胚发生的频率,促进其花药植株再生体系的优化,以橡胶树品种‘云研73-477’、‘热垦525’和‘云研77-2’的花药为外植体,研究不同浓度的TDZ对愈伤组织诱导和细胞胚胎发生能力的影响。结果表明,在愈伤组织诱导实验中,MS+TDZ 0.2 mg/L+ 6-BA 1 mg/L+2,4-D 1.5 mg/L+NAA 0.5 mg/L+椰子水50 mL/L+蔗糖70 g/L效果最好,诱导率均达到最大,分别为98%、98.67%、28.6%,当TDZ浓度逐步升高时,其诱导率又有所下降。在体细胞胚诱导实验中,愈伤组织诱导阶段加入TDZ对体细胞胚的发生有抑制作用;将愈伤组织转移到不含TDZ的培养基中继代增殖后,体细胞胚的诱导率随继代次数的增加呈上升趋势;愈伤诱导阶段使用的TDZ浓度越低越利于体细胞胚的分化;在体胚诱导阶段以TDZ代替KT后,体细胞胚诱导率较低,且分化出的体胚为畸形胚。这表明TDZ虽然有利于愈伤组织的产生和生长,但不利于体细胞胚的发生。

关键词: 江淮地区, 江淮地区

Abstract: The objectives of this study were to improve the frequencies of callus induction and somatic embryogenesis of desirable rubber clones, and optimize the anther plantlets regeneration system. The effects of different concentrations of TDZ on callus induction and somatic embryogenesis were conducted by using anthers of Hevea brasiliensis from 3 clones:‘Yunyan 73-477’,‘Reken 525’and‘Yunyan 77-2’. The results indicated that the optimizing medium for callus induction was MS+TDZ 0.2 mg/L+6-BA 1 mg/L+2,4-D 1.5 mg/ L+NAA 0.5 mg/L+ coconut water 50 mL/L+sucrose 70 g/L. All 3 clones showed highest callus induction rates in this medium, 98%, 98.67% and 28.6% respectively. The induction rate of callus was gradually declining with increasing concentration of TDZ. In the somatic embryogenesis induction test, TDZ restrained the callus regeneration when it existed in callus inducing period. When the callus was then transferred to the subculture medium without TDZ, the induction rate of somatic embryogenesis was rising with the increasing subculture times. And lower TDZ concentration was more favorable for the somatic embryo induction in callus inducing period. The induction rate of somatic embryogenesis was lower and there was no normal embryo when using TDZ instead of KT in the somatic embryogenesis inducing period. These showed that the TDZ was favorable for the callus induction and growth, but not for the somatic embryogenesis induction.