| 潘登浪 1，2，邹积鑫 1，2，曾宪海 1，2，林位夫 1，2，Khengtuan Chean1.油棕细胞悬浮培养及植株再生技术[J].广东农业科学,2019,46(2):59-65 |
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油棕细胞悬浮培养及植株再生技术 |
| Suspension Culture and Plant Regeneration of Oil Palm (Elaeis guineensis Jacq.) |
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| DOI：10.16768/j.issn.1004-874X.2019.02.009 |
| 中文关键词: 油棕 胚性愈伤组织 悬浮培养 体细胞胚 油料作物 |
| 英文关键词: oil palm embryogeneses callus suspension culture somatic embryo oil crop |
| 基金项目:海南省自然科学基金（317277）；海南省科技合作重点研发计划项目（ZDYF2018223） |
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| 摘要点击次数: 2526 |
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| 中文摘要: |
| 【目的】建立油棕胚性体细胞悬浮培养植株再生技术体系，为油棕优株无性系种植材料高效繁育、
细胞生物学研究、生物技术育种研究等提供技术平台和参考。【方法】以源于成龄油棕芯叶的瘤状愈伤组织为
材料，采用不同质量浓度 2,4-D、麦草畏与德夸霉素组合诱导易碎胚性愈伤组织，分析不同激素质量浓度及其
组合对油棕体细胞悬浮系增殖与后续体胚发育的影响。【结果】0.1 mg/L 德夸霉素与一定浓度 2,4-D 或麦草畏
组合，促进胚性愈伤组织形成，0.1 mg/L 德夸霉素与 1 mg/L 麦草畏组合诱导易碎胚性愈伤组织较佳，诱导率为
28.67%；用生长旺盛、质地松散的易碎胚性愈伤组织悬浮培养 21 d 可建立生长状态较均一、分散性好的悬浮
细胞系；0.3 mg/L 德夸霉素与 1 mg/L 麦草畏组合较有利于悬浮细胞系增殖和后续体胚发育；将悬浮细胞系转到
MS+ 谷氨酰胺 100 mg/L + 蔗糖 30g/L 培养基上，于 28（±2）℃、2 000 lx 下进行胚诱导培养，每两周更换新培
养基 1 次，培养 60 d 可获得直径大于 1 mm 的胚，最高每瓶 295 粒；将大于 1 mm 的胚转到固体培养基培养可
获得再生植株。【结论】适当质量浓度德夸霉素与 2,4-D 或麦草畏组合，促进油棕芯叶瘤状愈伤组织形成易碎
胚性愈伤组织；用生长旺盛、质地松散的易碎胚性愈伤组织悬浮培养 21 d 可建立生长状态较均一、分散性好的
悬浮细胞系；初步发现含适当质量浓度德夸霉素与麦草畏液体培养基利于悬浮细胞系增殖和后续体胚发育。 |
| 英文摘要: |
| 【Objective】 The objective was to establish a plant regeneration technology system for the suspension
culture of oil palm embryonic somatic cells, and to provide a technical platform and references for the efficient
breeding, cell biology research and biotechnology breeding research of the clonal planting materials of excellent oil
palm strains. 【Method】 Nodular callus derived from mature oil palm core leaves was used as starting materials for
suspension cultures.The effects of different hormone concentrations and their combinations on the proliferation of oil palm suspension cell lines and the development of subsequent somatic embryos were analyzed by using the combination
of different concentrations of 2, 4-D, Dicamba and Decoyinine to induce friable embryogenic callus. 【Result】The
combinations of Decoyinine (0.1 mg/L) with a certain concentration range of 2,4-D or Dicamba could promote the formation
of embryogenic callus. The combination of 0.1 mg/L Decoyinine and 1 mg/L Dicamba showed the best induction effect on
friable embryogenic callus, with an induction rate of 28.67%. A suspension cell line with uniform growth state and good
dispersion could be established after suspension culture of friable embryogenic callus with vigorous growth and loose texture
for 21 days. The combination of 0.3 mg/L Decoyinine and 1 mg/L Dicamba was beneficial to the proliferation of suspension
cell lines and the subsequent somatic embryos development. The suspension cells were transferred to embryo induction
medium (MS + glutamine 100 mg/L + sucrose 30 g/L), and cultured under 2 000 lx illumination at 28(±2)℃ . The culture
medium was replaced every two weeks. After 60 days of culture, embryos with the diameter larger than 1mm were obtained,
with a maximum of 295 embryos per flask. Regenerated plants could be obtained by transferring the embryos with the
diameter larger than 1mm to solid medium. 【Conclusion】 The combination of appropriate concentration of Decoyinine with
2,4-D or Dicamba could promote the formation of friable embryogenic callus from nodular callus of oil palm core leaves. A
suspension cell line with uniform growth state and good dispersity could be established after suspension culture of friable
embryogenic callus with vigorous growth and loose texture for 21 days. It was found that the liquid medium containing
appropriate concentration of Decoyinine and Dicamba was beneficial to the proliferation of suspension cell line and the
development of subsequent somatic embryos. |
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