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

Sun, Yongdong; Luo, Weirong; Li, Zhenxia; Lǐ, Yànhuá; Ni, Lei

doi:10.17957/ijab/15.0334

Cited by 7 publications

(5 citation statements)

References 18 publications

(27 reference statements)

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“…Wang et al [39] found that explants pre-cultured for 1 day grew more resistant shoots than explants used for direct infection, and the rate of resistant shoots increased from 11.1% to 27.3%; however, as the preculture time was prolonged, the cell division ability was weakened, and the transformation rate also began to decline. Similar conclusions were obtained by Sun et al [41]: with the progress of pre-culture, the bud induction rate first increased and then decreased, and number of shoots per explant (90.01% and 1.5, respectively). Fan et al [54] reported that compared with 1-2 d of pre-cultivation, a higher number of regenerated buds and resistant buds could be obtained in 3-7 d, considering that an excessive pre-culture time will lead to difficulties in later positive screening.…”

Section: Pre-culture Periodsupporting

confidence: 89%

“…However, it was also observed that some materials did not take root well with MS medium alone [29]; therefore, auxin and its analogs were often added to the medium in some studies. These could promote the rooting of shoots and accelerate the rooting process of adventitious shoots [27,41]. In addition, thidiazuron is an exogenous hormone commonly used in the culture of unfertilized cucumber cells and ovules and plays a key role in inducing the stage of explant callus formation to somatic embryogenesis [42].…”

Section: Exogenous Hormonementioning

confidence: 99%

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Recent Progress in the Regeneration and Genetic Transformation System of Cucumber

et al. 2022

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Cucumber (Cucumis sativus L.), belonging to the gourd family (Cucurbitaceae), is one of the major vegetable crops in China. Conventional genetic breeding methods are ineffective for improving the tolerance of cucumber to various environmental stresses, diseases, and pests in the short term, but bio-engineering technologies can be applied to cucumber breeding to produce new cultivars with high yield and quality. Regeneration and genetic transformation systems are key technologies in modern cucumber breeding. Compared with regeneration systems, genetic transformation systems are not yet fully effective, and the low efficiency of genetic transformation is a bottleneck in cucumber cultivation. Here, we systematically review the key factors influencing the regeneration and genetic transformation of cucumber plants, including the selection of genotype, source of explants and forms of exogenous hormones added to the medium, the methods of transgene introduction and co-cultivation, and selection methods. In addition, we also focus on recent advances in the study of molecular mechanisms underlying important agronomic traits using genetic transformation technology, such as fruit length, fruit warts, and floral development. This review provides reference information for future research on improvements in cucumber varieties.

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Section: Pre-culture Periodsupporting

confidence: 89%

Section: Exogenous Hormonementioning

confidence: 99%

Recent Progress in the Regeneration and Genetic Transformation System of Cucumber

et al. 2022

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“…The Agrobacterium tumefaciens-mediated genetic transformation system is currently the most desirable transformation method for dicotyledonous plants. Its transformation efficiency is often influenced by strain type ( Jaiwal et al., 2001 ; Yadav et al., 2014 ; Chen et al., 2018 ), explant pre-culture time ( Miguel and Oliveria, 1999 ; Yao et al., 2013 ; Verma et al., 2014 ; Mishra et al., 2016 ), infection time ( Sgamma et al., 2015 ; Li et al, 2018b ; Huang et al., 2017 ), infection concentration ( Coolado et al., 2015 ; Sun et al., 2017 ) co-culture time ( Tao and Ling, 2006 ; Wu et al., 2017 ), and AS concentration and addition method ( Singh et al., 2017 ). Different plants or different parts of the same plant may require unique transformation conditions.…”

Section: Discussionmentioning

confidence: 99%

An effective method for establishing a regeneration and genetic transformation system for Actinidia arguta

Yao,

Kong,

Lei

et al. 2023

Front. Plant Sci.

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The all-red A. arguta (Actinidia arguta) is an anthocyanin-rich and excellent hardy fruit. Many studies have focused on the green-fleshed A. arguta, and fewer studies have been conducted on the all-red A. arguta. Here we reported a regeneration and Agrobacterium-mediated transformation protocol by using leaves of all-red A. arguta as explants. Aseptic seedling leaves of A. arguta were used as callus-inducing materials. MS medium supplemented with 0.3 mg·L-1 2,4-D and 1.0 mg·L-1 BA was the optimal medium for callus induction of leaves, and medium supplemented with 3 mg·L-1 tZ and 0.5 mg·L-1 IAA was optimal for adventitious shoot regeneration. The best proliferation medium for adventitious buds was MS + 1.0 mg·L-1 BA + 0.3 mg·L-1 NAA. The best rooting medium was 1/2MS + 0.7 mg·L-1 IBA with a 100% rooting rate. For the red flesh hardy kiwi variety ‘Purpurna Saduwa’ (A. arguta var. purpurea), leaves are receptors for Agrobacterium (EHA105)-mediated transformation. The orthogonal experiment was used for the optimization of each genetic transformation parameter and the genetic transformation of the leaves was 21% under optimal conditions. Our study provides technical parameters for applying genetic resources and molecular breeding of kiwifruit with red flesh.

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“…However, these parameters were different in various literatures and also lack of enough details in the existing studies. All of these reasons leaded to browning, Agrobacterium pollution, vitrification shoots, low efficiency of genetic transformation [24], gene expression and genetic instability, which still perplex researchers. In order to improve the efficiency of genetic transformation, the effects of different concentrations of kanamycin on the regeneration of cotyledonary nodes by applying it in different transformation periods were analyzed in the present study.…”

Section: Introductionmentioning

confidence: 99%

Improved the Agrobacterium tumefaciens-mediated transformation of cucumber by a modified the using of antibiotics and acetosyringone

Chai¹,

Du²,

Fan³

et al. 2020

Preprint

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Background: Cucumber (Cucumis sativus) is one of the most important vegetable crops in the world. As conventional breeding of cucumber is very challenging, genetic engineering is an alternative option to introduce important traits such as enhanced stress resistance and nutritional value. However, the efficiency of the transformation system depends on genotypes, transformation conditions, selection agents, etc. This study aims to speed up the process of Agrobacterium-mediated transformation of cucumber. ‘ Xintai mici ’, a very popular and typical north China-type cucumber variety, was transformed with Agrobacterium GV3101. The strain carried pCAMBIA2300s plasmid, a double vector with the marker gene of neomycin phosphotransferase II ( npt II). Results: The research results indicated that cefotaxime sodium was suitable for inhibiting Agrobacterium in the stage of screening and bud elongation. Timentin was best used during rooting stage. Furthermore, 25 mg/L kanamycin was used in the early stage of screening and increased to 50 mg/L for further screening. At the bud elongation and rooting stage, 75 and 100 mg/L kanamycin was used respectively to improve the screening efficiency. In order to obtain the highest regeneration frequency of resistant buds, 50, 150, and 100 μM acetosyringone were added in the pre-culture medium, infection solution, and co-culture medium respectively. To confirm the presence of the transgenes, DNA from npt II transgenic cucumber plants was analyzed by polymerase chain reaction after transplanting resistant regenerated plants. Conclusions: We finally achieved an 8.1% conversion, which was among the highest values reported until date using cucumber ‘ Xintai mici ’. Thus an effective protocol for Agrobacterium tumefaciens -mediated genetic transformation of cucumber was optimized.

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

Cited by 7 publications

References 18 publications

Recent Progress in the Regeneration and Genetic Transformation System of Cucumber

Recent Progress in the Regeneration and Genetic Transformation System of Cucumber

An effective method for establishing a regeneration and genetic transformation system for Actinidia arguta

Improved the Agrobacterium tumefaciens-mediated transformation of cucumber by a modified the using of antibiotics and acetosyringone

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