Agrobacterium-mediated transformation of cucumber (Cucumis sativus L.) using a sense mitogen-activated protein kinase gene (CsNMAPK)

Wang, Jing; Zhang, Shoujie; Wang, Xiao; Wang, Lina; Wang, Xiufeng; Shi, Qinghua; Wei, Min; Yang, Fang

doi:10.1007/s11240-012-0266-y

Cited by 20 publications

(14 citation statements)

References 38 publications

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“…It was observed that 2 d of pre-cultivation and 2 d of co-cultivation were very effective in producing the highest cucumber regeneration frequency and number of shoots in cucumber. These findings of present study are in agreement with those found by Cardoza and Stewart (2003), Jonoubi et al (2005), Khan et al (2010), Wang et al (2013) and Khan et al (2013). The explants co-cultured for 2 d showed the highest regeneration frequency and number of shoots in current study.…”

Section: Discussionsupporting

confidence: 94%

“…150 mg L -1 of kanamycin was used to select transformants by Wang et al (2013), 200 mg L -1 of kanamycin was used by Oliveira et al (2011), Mishra et al (2013), and Alvarez and Ordás (2013). In present study, transformed cucumber lines were successfully selected at 100 mg L -1 of kanamycin.…”

Section: Discussionmentioning

confidence: 80%

“…The explants co-cultured for 2 d showed the highest regeneration frequency and number of shoots in current study. Previously, co-cultivation for 2 d was reported by Wang et al (2013). In contrast, Mishra et al (2013) reported that explants co-cultivated for 5 d showed the maximum transformation efficiency in rice.…”

Section: Discussionmentioning

confidence: 94%

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Establishment of a High-efficiency Genetic Transformation System of Cucumber (Cucumis sativus) using Csexpansin 10 (CsEXP10) Gene

Sun¹,

Luo²,

Li³

et al. 2017

IJAB

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Cucumber is one of the most important vegetable crops in the world. To establish an efficient genetic transformation system in cucumber, the role of different factors such as pre-cultivation time, acetosyringone concentration, infection time and cocultivation time that can influence the transformation, was evaluated. In addition, Csexpansin 10 (CsEXP10) gene was transformed into the cucumber genome to produce transgenic lines. The results showed that various optimal parameters such as 100 mg L -1 of kanamycin concentration for selection of transformants, 2 d of pre-cultivation time, 100 μmol L -1 of acetosyringone concentration, 15 min of infection time and 2 d of co-cultivation time were obtained using cotyledonary node explants. The rooting frequency observed on Murashige and Skoog (MS) medium supplemented with 0.2 mg L -1 indole acetic acid and 400 mg L -1 cefotaxime was found to be 100.00%. The positive transgenic cucumber lines were identified using PCR analysis and GUS staining assay. It suggests that the genetic transformation system developed using cotyledonary node explants is efficient and successful in cucumber.

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Section: Discussionsupporting

confidence: 94%

Section: Discussionmentioning

confidence: 80%

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Establishment of a High-efficiency Genetic Transformation System of Cucumber (Cucumis sativus) using Csexpansin 10 (CsEXP10) Gene

Sun¹,

Luo²,

Li³

et al. 2017

IJAB

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“…Previous studies have shown a high regeneration frequency with cotyledon nodes as explants 36 , and we also found that breaking off the hypocotyl to detach the cotyledon, rather than cutting it, is important to obtain high regeneration frequency. However, care needs to be taken to cut off the growing point of the seedlings to prevent negative plants growing.…”

Section: Explant Preparationsupporting

confidence: 55%

A Protocol for Agrobacterium-mediated Transformation of Cucumber (Cucumis sativus L.) from cotyledon explants

X¹,

Ma²,

Shan³

et al. 2017

Protocol Exchange

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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.

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“…Cucumber is an economically important crop, and is widely cultivated throughout the world (Wang et al 2013). The species belongs to the group of the most popular vegetables worldwide (Nishibayashi et al 1996).…”

Section: Introductionmentioning

confidence: 99%

Production of transgenic cucumber expressing phytoene synthase-2A carotene desaturase gene

et al. 2016

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This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.Abstract The objectives of this study were to 1) evaluate the efficiency of the protocol of Agrobacterium-mediated transformation of cucumber to introduce phytoene synthase-2a carotene desaturase (PAC genes); 2) demonstrate the integration of PAC genes into the genome of putative transgenic cucumber based on growth on selection medium, PCR and Southern analysis; 3) evaluate the expression of PAC genes in transgenic cucumber based on the analysis of RT-PCR and Northern blot hybridization. Out of 5,945 cotyledonary-node explants inoculated with Agrobacterium, 65 (1.1%) explants produced 238 shoots. Integration of PAC genes into the genome of the cucumber was demonstrated based on the analysis of gDNA-PCR, 21 out of the 238 plants regenerated; while 6 plants proved positive for Southern blot hybridization. Transgene expression was demonstrated based on analysis of RT-PCR, 6 plants proved positive out of the 6 plants analyzed; while 4 plants out of 6 proved positive during Northern blot hy-bridization. This study successfully demonstrated the production of transgenic cucumber, integration, and expression of the PAC gene in cucumber.

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Agrobacterium-mediated transformation of cucumber (Cucumis sativus L.) using a sense mitogen-activated protein kinase gene (CsNMAPK)

Cited by 20 publications

References 38 publications

Establishment of a High-efficiency Genetic Transformation System of Cucumber (Cucumis sativus) using Csexpansin 10 (CsEXP10) Gene

Establishment of a High-efficiency Genetic Transformation System of Cucumber (Cucumis sativus) using Csexpansin 10 (CsEXP10) Gene

A Protocol for Agrobacterium-mediated Transformation of Cucumber (Cucumis sativus L.) from cotyledon explants

Production of transgenic cucumber expressing phytoene synthase-2A carotene desaturase gene

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