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黄瓜胚性愈伤组织的诱导保存和再生
薛婉钰1, 刘娜1, 苑鑫,等1
西北农林科技大学 园艺学院,陕西省蔬菜工程技术研究中心
摘要:
【目的】对黄瓜胚性愈伤组织的诱导保存和再生进行研究,为黄瓜高频率遗传转化奠定基础。【方法】以欧洲温室型黄瓜自交系14-1子叶节为外植体,在MS培养基上附加1.5 mg/L 2,4-D,进行25 d的胚性愈伤组织诱导培养后,取胚性愈伤组织在添加30,60,90,100,110,120,130,140和150 g/L蔗糖及1.5 mg/L 2,4-D的MS培养基进行继代培养,每30 d继代1次,观察胚性愈伤组织的褐变情况及胚性分化能力,并用电子天平在超净工作台中记录胚性愈伤组织质量的变化。继代培养60 d后,将保存的胚性愈伤组织和体细胞胚移至含1.5 mg/L 2,4-D的MS培养基上,待出现体细胞胚后移至MS培养基进行萌发,观察再生小植株的生长情况。【结果】将欧洲温室型黄瓜自交系14-1的子叶节,接种到附加1.5 mg/L 2,4-D的MS培养基上进行诱导培养后,子叶节一端的愈伤组织集中聚集于下胚轴处,之后有黄色胚性愈伤组织产生。在继代培养过程中,当培养基中添加的蔗糖为60~150 g/L时,胚性愈伤组织能保持胚性愈伤状态达60 d。之后将继代培养60 d后的胚性愈伤组织转接至附加1.5 mg/L 2,4-D的MS培养基上,在蔗糖质量浓度为60 g/L条件下保存的胚性愈伤组织可诱导出正常胚状体,且能形成健康小植株。【结论】由黄瓜子叶节诱导出的胚性愈伤组织可在MS+60 g/L蔗糖的培养基上保存达60 d,之后能正常萌发形成胚状体,进而形成正常小植株。
关键词:  黄瓜;遗传转化  胚性愈伤组织;离体保存
DOI:
分类号:
基金项目:国家自然科学基金项目(32072562;32272748)
Induction and preservation of cucumber embryogenic callus
XUE Wanyu,LIU Na,YUAN Xin,et al
Abstract:
【Objective】This study induced and preserved embryogenic callus of cucumber cotyledon nodes as explants to provide basis for high-frequency genetic transformation system of cucumber.【Method】The cotyledon nodes of the inbred line 14-1 of European ecotype were inoculated on the MS medium supplemented with 1.5 mg/L 2,4-D.After cotyledon nodes were inoculated on callus induction medium for 25 days,the induced embryogenic calli were transferred to MS medium supplemented with 30,60,90,100,110,120,130,140 and 150 g/L sucrose and 1.5 mg/L 2,4-D,and they were subcultured once every 30 days.The differentiation ability of embryogenic calli and browning were observed,and the quality changes of embryogenic calli were recorded in clean bench.The preserved embryogenic calli and somatic embryos were transferred to MS medium containing 1.5 mg/L 2,4-D for 60 days to observe the growth status of regenerated plantlets.【Result】After inoculating cotyledon nodes of European greenhouse type cucumber inbred line 14-1 onto MS medium supplemented with 1.5 mg/L 2,4-D for induction culture,the embryogenic calli concentrated in the hypocotyl and they became yellow afterwards.The embryogenic calli of the high-sugar medium in sucrose concentrations of 60-150 g/L might be preserved for 60 days.After the embryogenic calli were transferred to MS medium supplemented with 1.5 mg/L 2,4-D,somatic embryos can be induced when preserved at sucrose concentration of 60 g/L and healthy plantlets were formed.【Conclusion】The embryogenic calli induced from cucumber cotyledon nodes can be preserved on the MS medium with 60 g/L sucrose for 60 days,and they were able to germinate normally to form embryos and normal plantlets.
Key words:  cucumber  genetic tromsformation  embryogenic callus  in vitro conservation