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Method Article
zhenxian zhang
Beijing Key Laboratory of Growth and Developmental Regulation for Protected Vegetable Crops, College of Horticulture, China Agricultural University, Beijing, P. R. China.
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Genetic transformation is central to the discovery, delivery, and functional analysis of genes, and for associated crop improvement, and yet is technically challenging in many plant species. As an example, cucumber (Cucumis sativus L.) is a model species for the study of phloem characteristics, raffinose family oligosaccharide (RFO) metabolism and sex determination, but methods for its genetic transformation have generally had low efficiency, thereby limiting basic and applied studies. Here, we describe a rapid and efficient protocol for Agrobacterium-mediated transformation of cucumber cotyledon explants, using vacuum infiltration. The transformation efficiency of the regenerated plants was as high as 54%, and the final positive plantlet transformation efficiency was approximately 26% of the total number of infected explants and this is obviously higher than previously published protocols. Transgenic plantlets can be obtained in 3 to 4 months and the transgenic T1 seeds generated in the subsequent 3 to 4 months after self-pollination. Using this protocol, we have obtained more than 600 transgenic cucumber lines. This protocol is of great importance for studies of cucumber and of other Cucurbitaceae species, since it enables gene functional analysis, and so opens a pipeline for rapid cucumber variety improvement.
Keywords
cucumber, Agrobacterium-mediated, transformation
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Xin Li and Si Ma contributed equally to this work. The authors declare no competing financial interests.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.jpg
Supplementary Figure S1 Supplementary Figure S1 ∣ Combined effect of different hormones and base media on cucumber regeneration using the Xintaimici and Chinese long 461 genotypes. A, Combined effect of ABA and BAP on cucumber regeneration using Xintaimici after 15 days of shoot regeneration culturing. B, Combined effect of MS and B5 base media on cucumber regeneration using 1.0 mg/l ABA and different BAP concentrations and the Chinese long 461 genotype after 20 days of shoot regeneration culturing.
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Posted 19 Sep, 2017
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Associated Publications
Down-Regulating CsHT1, a Cucumber Pollen-Specific Hexose Transporter, Inhibits Pollen Germination, Tube Growth, and Seed Development
by Jintao Cheng, Zhenyu Wang, Fengzhen Yao, +4
PLANT PHYSIOLOGY (28 July, 2017)
Antisense suppression of cucumber ( Cucumis sativus L.) sucrose synthase 3 ( CsSUS3 ) reduces hypoxic stress tolerance
by HONGYUN WANG, XIAOLEI SUI, JINJU GUO, +5
A GAMYB homologue CsGAMYB1 regulates sex expression of cucumber via an ethylene-independent pathway
by Yan Zhang, Xiaolan Zhang, Bin Liu, +6
HANABA TARANU regulates the shoot apical meristem and leaf development in cucumber ( Cucumis sativus L.)
by Lian Ding, Shuangshuang Yan, Li Jiang, +7
Suppression of cucumber stachyose synthase gene (CsSTS) inhibits phloem loading and reduces low temperature stress tolerance
by Jianguo Lü, Xiaolei Sui, Si Ma, +3
Plant Molecular Biology (29 July, 2017)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
zhenxian zhang
Beijing Key Laboratory of Growth and Developmental Regulation for Protected Vegetable Crops, College of Horticulture, China Agricultural University, Beijing, P. R. China.
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 19 Sep, 2017
Community comments: 2
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Genetic transformation is central to the discovery, delivery, and functional analysis of genes, and for associated crop improvement, and yet is technically challenging in many plant species. As an example, cucumber (Cucumis sativus L.) is a model species for the study of phloem characteristics, raffinose family oligosaccharide (RFO) metabolism and sex determination, but methods for its genetic transformation have generally had low efficiency, thereby limiting basic and applied studies. Here, we describe a rapid and efficient protocol for Agrobacterium-mediated transformation of cucumber cotyledon explants, using vacuum infiltration. The transformation efficiency of the regenerated plants was as high as 54%, and the final positive plantlet transformation efficiency was approximately 26% of the total number of infected explants and this is obviously higher than previously published protocols. Transgenic plantlets can be obtained in 3 to 4 months and the transgenic T1 seeds generated in the subsequent 3 to 4 months after self-pollination. Using this protocol, we have obtained more than 600 transgenic cucumber lines. This protocol is of great importance for studies of cucumber and of other Cucurbitaceae species, since it enables gene functional analysis, and so opens a pipeline for rapid cucumber variety improvement.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Xin Li and Si Ma contributed equally to this work. The authors declare no competing financial interests.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.jpg
Supplementary Figure S1 Supplementary Figure S1 ∣ Combined effect of different hormones and base media on cucumber regeneration using the Xintaimici and Chinese long 461 genotypes. A, Combined effect of ABA and BAP on cucumber regeneration using Xintaimici after 15 days of shoot regeneration culturing. B, Combined effect of MS and B5 base media on cucumber regeneration using 1.0 mg/l ABA and different BAP concentrations and the Chinese long 461 genotype after 20 days of shoot regeneration culturing.
Loading...
Associated Publications
Down-Regulating CsHT1, a Cucumber Pollen-Specific Hexose Transporter, Inhibits Pollen Germination, Tube Growth, and Seed Development
by Jintao Cheng, Zhenyu Wang, Fengzhen Yao, +4
PLANT PHYSIOLOGY (28 July, 2017)
Antisense suppression of cucumber ( Cucumis sativus L.) sucrose synthase 3 ( CsSUS3 ) reduces hypoxic stress tolerance
by HONGYUN WANG, XIAOLEI SUI, JINJU GUO, +5
A GAMYB homologue CsGAMYB1 regulates sex expression of cucumber via an ethylene-independent pathway
by Yan Zhang, Xiaolan Zhang, Bin Liu, +6
HANABA TARANU regulates the shoot apical meristem and leaf development in cucumber ( Cucumis sativus L.)
by Lian Ding, Shuangshuang Yan, Li Jiang, +7
Suppression of cucumber stachyose synthase gene (CsSTS) inhibits phloem loading and reduces low temperature stress tolerance
by Jianguo Lü, Xiaolei Sui, Si Ma, +3
Plant Molecular Biology (29 July, 2017)