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黄瓜胚状体途径再生体系的建立
魏文霞1, 刘苗苗1, 张婷婷,等1
西北农林科技大学 园艺学院
摘要:
【目的】建立黄瓜高频胚状体再生体系,为黄瓜遗传转化体系的完善奠定基础。【方法】以黄瓜品种津研四号、长春密刺、Q8和14-1为材料,研究不同基因型材料愈伤组织的诱导情况;分别取14-1的子叶、子叶节、下胚轴和成熟胚为外植体,研究不同外植体类型的愈伤组织和胚状体产生情况;以14-1子叶节为外植体,研究不同激素及其质量浓度配比对黄瓜胚状体诱导的影响。【结果】4个不同基因型中,欧洲温室型黄瓜种质14-1可诱导出胚状体。在4种外植体类型中,14-1的子叶节可以高效诱导出愈伤组织和胚状体,其诱导率分别为100.0%和34.1%。激素及其质量浓度组合也会影响胚状体的诱导,在愈伤组织诱导阶段,以MS+1.5 mg/L 2,4-D为培养基对胚性愈伤组织的诱导效果最好,其对14-1子叶节的诱导率为100%;在分化培养阶段,MS+0.1 mg/L 2,4-D+0.5 mg/L KT+0.2 mg/L IAA有利于体细胞胚的发生,14-1的出胚率为62.5%。显微形态观察发现,黄瓜体细胞胚发育过程包括球形胚、心形胚、鱼雷胚和子叶期胚等时期,这些胚可以发育成正常小植株。【结论】14-1的子叶节经诱导培养基MS+1.5 mg/L 2,4-D和分化培养基MS+0.1 mg/L 2,4 D+0.5 mg/L KT+0.2 mg/L IAA培养后,可高效产生胚状体,并可萌发成正常植株。
关键词:  黄瓜  愈伤组织  胚状体  体细胞胚发育  组织培养
DOI:
分类号:
基金项目:国家自然科学基金项目(31572143,31772335)
Establishment of cucumber somatic embryo regeneration system
WEI Wenxia,LIU Miaomiao,ZHANG Tingting,et al
Abstract:
【Objective】This study established an efficient regeneration system of cucumber via somatic embryogenesis to provide basis for the genetic transformation system.【Method】Four cultivars cucumber such as “Jinyansihao”,“Changchunmici”,“Q8” and “14-1” were used to investigate the effection of different genotypes on callus and embryo induction.Different explants types such as cotyledonary node,cotyledon,hypocotyledonary axis and mature embryo from 14-1 cucumber were obtained to study the effection of explants types.Cotyledonary node of 14-1 were used to investigate the callus and embryo induction supplied with different concentration of hormone.【Result】It showed that genotye 14-1 was able to be induced somatic embryos among the four genotypes.The cotyledon,cotyledon nodes and hypocotyls of 14-1 were induced callus efficiently on the first stage,however,only cotyledon nodes in four kind of explants was able to be induced embryogenic callus and embryos,the induction rates were 100.0% and 34.1%,respectively.The different hormone combinations also affected the percentage of embryo induction,and it showed that the suitable hormone combination for embryo induction of 14-1 was on the MS mediums supplying with 1.5 mg/L 2,4-D to induce callus first and then the embryo was induced on MS medium supplying with 0.1 mg/L 2,4-D,0.5 mg/L KT and 0.2 mg/L IAA,and percentage of embryo formation was 62.5%.The globular embryo,heart-shaped embryo,torpedo-shaped embryo and cotyledonary embryo were observed under a stereoscopic microscope.These embryo was able to grow into plants.【Conclusion】The cotyledon nodes of 14-1 can produce somatic embryos efficiently on the optimal embryogenic callus induced medium(MS medium supplemented with 1.5 mg/L 2,4-D) and embryos induced medium(MS medium supplying with 0.1 mg/L 2,4-D,0.5 mg/L KT and 0.2 mg/L IAA).These somatic embryos can germinate and grow into plants.
Key words:  Cucumis Sativus L.  callus  embryoid  somatic embryogenesis  tissue culture