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。
[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 Dicamha showed the best induction effect on friable embryogenic callus, with an indution 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 1 x illumination at 28 (± 2)℃ The culture medium was replaced every two weeks. After 60 days of culture, embryos with the diameter larger than 1 mm were obtained,with a maximum of 295 embryos per flask. Regenerated plants could be obtained by transferring the embryos with the diameter larger than 1 mm to solid medium. [Conclusion ]The combination of appropriate concentration of Decoyine with2,4-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 Deoyinine and Dicamba was beneficial to the proliferation of suspension cell line and the development of subsequent somatic embryos.
作者
潘登浪
邹积鑫
曾宪海
林位夫
Khengtuan Chean
PAN Denglang;ZOU Jixin;ZENG Xianhai;LIN Weifu;Khengtuan Cheah(Rubber Research Institute,CATAS/State Key Laboratory Incubation Base for Cultivation&Physiology of Tropical Crops/Engineering Center of Speedy Propagation for Tropical Woody Plants,CATAS,Haikou 571701,China;Germplasm Repository of Oil Palm,Ministry of Agriculture and Rural Affairs/Danzhou Investigation&Experiment Station of Tropical Crops,Ministry of Agriculture and Rural Affairs,Danzhou 571737,China)
出处
《广东农业科学》
CAS
2019年第2期59-65,173,共8页
Guangdong Agricultural Sciences
基金
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)
关键词
油棕
胚性愈伤组织
悬浮培养
体细胞胚
油料作物
oil palm
embryogeneses callus
suspension culture
somatic embryo
oil crop
作者简介
潘登浪(1980—),男,壮族,硕士,农艺师,研究方向为热带经济作物组织与细胞培养、种质资源保护与利用,E-mail:pandenglang@163.com;通信作者:林位夫(1955—),男,研究员,研究方向为热带作物栽培,E-mail:rubberl@163.com.
