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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (25): 15-19.

所属专题： 生物技术； 园艺

• 林学 园艺 园林 • 上一篇    下一篇

不同因素对橡胶树内珠被愈伤组织诱导和体细胞胚发生的影响

孙爱花 黄天带 周权男 戴雪梅 华玉伟 黄华孙   

  • 收稿日期:2012-05-11 修回日期:2012-05-23 出版日期:2012-09-05 发布日期:2012-09-05

Effects of Different Factors on Callus Induction and Somatic Embryogenesis of Inner Integument from Hevea Brasiliensis

  • Received:2012-05-11 Revised:2012-05-23 Online:2012-09-05 Published:2012-09-05

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摘要：

为了优化橡胶树内珠被植株再生体系，促进其次生体胚技术和转基因研究的发展，以橡胶树品种‘热研7-33-97’和‘热研88-13’的幼果内珠被为材料，进行不同因素对橡胶树内珠被愈伤组织诱导和体细胞胚发生的研究。在添加不同浓度的2,4-D，外加80 g/L蔗糖、50 mg/L椰子水、5.0 mg/L AgNO3和2.3 g/L phytagel的MH培养基上诱导愈伤组织的产生。结果表明，当培养基中不添加2,4-D时，2个品种均不能诱导出愈伤组织，随着2,4-D浓度的增加，其愈伤诱导率不断增加，达到4.0 mg/L水平时，诱导率均达到最大，分别为88.28%、86.67%。当浓度逐步升高至6.0 mg/L时，其诱导率又有所下降。愈伤诱导后，将其转移到添加了不同浓度NAA的改良MS基本培养基上。结果发现，0.2 mg/L的NAA浓度更有利于橡胶树‘热研7-33-97’内珠被子叶胚、喇叭胚、杯状胚和多叶胚等胚的产生，但子叶胚出胚率也仅达4.00%，而低于或高于此浓度均不利于体细胞胚的产生。

关键词: 干燥基准, 干燥基准

Abstract:

In order to optimize the plant regeneration system of inner integument and promote the development of secondary somatic embryogenesis and transgenetic research of Hevea brasiliensis. Callus induction and somatic embryogenesis were conducted on different factors by taking inner integument of immature fruits from two clones Reyan 7-33-97 and Reyan 88-13 of Hevea brasiliensis. Calla induction were obtained on MH medium supplemented with 80 g/L sugar, 50 mg/L coconut water, 5.0 mg/L silver nitrate, 2.3 g/L phytagel and different concentrations of 2,4-D. The results showed that calla could not be inducted from two clones when the media had no 2,4-D. The induction rate of callus was gradually improved with the increasing of 2,4-D concentration. The rate was the highest, while the concentration reached 4.0 mg/L, respectively 88.28% and 86.67%. When the concentration of 2,4-D reached 6.0 mg/L, the rate was gradually declining. After the induction of callus, callus was transferred to modified MS medium supplemented with different concentrations of NAA. The results indicated that the embryoids of cotyledonary, cup-like, trumpet-like and leafy were easily produced with the concentration of 0.2 mg/L NAA from the inner integument of Reyan 7-33-97. The induction rate of cotyledonary embryoid reached only 4.00%. Somatic embryo was not beneficial to be produced when the concentration of NAA was lower or more than 0.2 mg/L.