| 摘要: |
| 【目的】筛选适宜于黄瓜体细胞胚发生的培养条件，为完善黄瓜胚状体再生途径及建立黄瓜优良的遗传转化受体系统奠定基础。【方法】以8份不同基因型黄瓜种质Q24、Q16、9930、No.26、Gy14、No.14、S63和16F7-2-8为材料，研究基因型对黄瓜胚状体诱导的影响。以胚状体诱导率及成苗率为考察指标，以Gy14和9930为材料，探讨苗态（子叶未展开，子叶即将展开，子叶展开）对黄瓜胚状体诱导的影响；以Gy14为材料，从子叶切割方式（不保留下胚轴及保留约1，2 mm下胚轴）、无菌苗培养阶段光照与否、愈伤组织诱导培养基中2,4-D的质量浓度（0.75，1.00，1.50，2.00 mg/L）以及胚性愈伤的诱导时长（20，30和60 d）等方面，对黄瓜胚状体再生体系进行优化。【结果】基因型选择结果显示，Gy14和9930比其他基因型黄瓜种质更易诱导出较明显的胚状体。与子叶即将展开和子叶已展开2种苗态相比，子叶未展开时，Gy14和9930的胚状体诱导率较高，分别可达39.3%和38.7%。子叶节保留约1 mm下胚轴处理的胚状体诱导率最高，可达85.2%。无菌苗培养阶段每天进行16 h光照和全天暗处理2种方式，对于胚状体诱导率的影响不显著。适宜胚状体诱导的2,4-D质量浓度为1.00 mg/L，胚状体诱导率为31.6%。与诱导20和60 d相比，外植体在诱导培养基上诱导30 d的愈伤组织胚状体诱导率和成苗率较高，分别为39.6%和65.6%。【结论】黄瓜Gy14的无菌苗处于子叶未展开、仍呈抱合状态时，对其采取保留约1 mm下胚轴的切割方式，置于含1.00 mg/L 2,4-D的培养基上培养30 d后，可较为高效地产生胚状体，并获得完整的植株。 |
| 关键词: 黄瓜 胚状体 愈伤组织 诱导率 成苗率 |
| DOI： |
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| 基金项目:国家自然科学基金项目（31772335）；西北农林科技大学试验示范站（基地）科技创新与成果转化项目（TGZX2020-20） |
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| Establishment and optimization of high frequency embryoid regeneration system of cucumber (Cucumis sativus L.) |
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LIU Na,XUE Wanyu,CHEN Panpan,et al
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| Abstract: |
| 【Objective】This study screened the suitable culture conditions for somatic embryogenesis of cucumber to provide basis for improving the embryoid regeneration pathway of cucumber and establishing excellent genetic transformation receptor system of cucumber.【Method】The effects of 8 different genotypes on embryoid induction of cucumber were probed using Q24,Q16,9930,No.26,Gy14,No.14,S63 and 16F7-2-8,and the effects of seedling state (cotyledons not unfolded,cotyledons ready for unfold and cotyledons unfolded) on embryoid induction of cucumber were investigated using Gy14 and 9930.The regeneration system of embryoid was optimized using Gy14 from the cutting ways of cotyledons (unreserved hypocotyl and 1 or 2 mm reserved hypocotyl),light or dark treatments of sterile seedlings, different concentrations of 2,4-D supplemented on induction medium (0.75,1.00,1.50 and 2.00 mg/L) and different induction times of embryogenic callus (20,30 and 60 d). The embryoid induction rate and seedling formation rate were used as indexes.【Result】The embryoids were induced more easily from Gy14 and 9930 than from other cucumber genotypes.The induction rate of unfold cotyledons was 39.3% for 9930 and 38.7% for Gy14,which were higher than that of the other two seedling states.The embryoid induction rate of cotyledons with about 1 mm hypocotyl was the highest of 85.2%.The embryoid induction rate was not significantly different when sterile seedlings were treated under light or dark conditions.The most suitable concentration of 2,4-D for embryoid induction was 1.00 mg/L with induction rate of 31.6%.The embryoid induction rate and seedling formation rate of 39.6% and 65.6% for the callus inducted for 30 d were higher than those inducted for 20 and 60 d.【Conclusion】The efficient embryoid production and complete plant obtaining method was inducing unfold cotyledons with 1mm hypocotyl and culturing on the medium supplemented with 1.00 mg/L 2,4-D for 30 days. |
| Key words: cucumber embryoid callus induction rate seeding formation rate |
