Improving the transformation efficiency of Cucurbita species: factors and strategy for practical application

Nanasato, Yoshihiko; Okuzaki, Ayako; Tabei, Yutaka

doi:10.5511/plantbiotechnology.13.0331a

Cited by 17 publications

(14 citation statements)

References 36 publications

(52 reference statements)

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“…Previous studies showed that the application of vacuuminfiltration treatment during A. tumefaciens infection significantly improved transformation efficiency in other recalcitrant plants (de Oliveira et al 2009;Bakshi et al 2011). Studies in other plants also showed that the use of filter-paper wicks as a culture support during the co-cultivation step improved transformation efficiency in some plants including recalcitrant crops such as wheat and kabocha squash (Cheng et al 2003;Ozawa 2009;Nanasato et al 2011;Nanasato et al 2013a;Nanasato et al 2013b). Therefore, the combined effect of vacuum infiltration and filter-paper wicks on Jatropha transformation was examined.…”

Section: Resultsmentioning

confidence: 97%

“…Recently, a technique of vacuum infiltration during A. tumefaciens infection has been introduced to improve transformation efficiency of recalcitrant crops including cowpea, citrus, cucumber, and Cucurbita species (de Oliveira et al 2009;Bakshi et al 2011;Nanasato et al 2011;Nanasato et al 2013a;Nanasato et al 2013b). Another technique developed recently involves the use of filter-paper wicks as a culture support during the co-cultivation period, which contributed to the efficient transformation of kabocha squash, rice, and wheat (Cheng et al 2003;Ozawa 2009;Nanasato et al 2011).…”

Section: Introductionmentioning

confidence: 99%

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Efficient genetic transformation of Jatropha curcas L. by means of vacuum infiltration combined with filter-paper wicks

Nanasato

Kido

Kato

et al. 2015

In Vitro Cell.Dev.Biol.-Plant

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Jatropha curcas L. (Jatropha) is a promising plant for the production of biodiesel in arid regions. To improve its productivity, it is important to understand the molecular functions of key genes in Jatropha, and to seek ways to modify important agronomic traits of Jatropha via molecular breeding. However, Jatropha is notorious for its recalcitrance to transformation. Thus, an improved protocol for efficient and reproducible Agrobacterium tumefaciens-mediated transformation is essential for molecular biology research and breeding in this species. Because resistance of Jatropha to A. tumefaciens infection is one of the key factors limiting transformation efficiency, we attempted to improve infection efficiency by comparing various co-culture conditions with the use of a β-glucuronidase (GUS) assay. The LBA4404 strain of A. tumefaciens was more efficient than the EHA105 strain for transformation of Jatropha. Vacuum infiltration of an A. tumefaciens suspension with Jatropha explants, combined with co-cultivation on filter-paper wicks moistened with coculture medium instead of on solid medium, significantly improved transformation efficiency. In these conditions, GUSpositive sectors were observed in 93.0±23.6% of infected explants. Moreover, a variant of the yellow fluorescent protein gene, Venus, was used as a visible marker, which proved effective for discrimination of candidate transgenic shoots from escape shoots. Transgene integration was confirmed by means of polymerase chain reaction and Southern hybridization analyses. Transgenic shoots were regenerated from 23% of explants from the vacuum-filter-paper treatment, which is sufficient for practical use.

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Section: Resultsmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Efficient genetic transformation of Jatropha curcas L. by means of vacuum infiltration combined with filter-paper wicks

Nanasato

Kido

Kato

et al. 2015

In Vitro Cell.Dev.Biol.-Plant

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“…The use of NPTII as a primer because of NPTII is a specific gene not owned by tomato plants (Meissner et al, 1997). In planta transformation has been successfully performed using the floral dip (Ratanasut et al, 2017;Bent 2000;Li et al, 2010;Rod-in et al, 2014;Narusaka et al, 2010;Zang et al, 2006 Clough andBent 1998), by soaking the flower of the plant in the liquid A. tumefaciens culture; the pin pricking (Razzaq et al, 2011;Jaganath et al, 2013;Seol et al, 2008), by pricking the transformation target with the needle smeared with A. tumefaciens; agro-injection (Bratic et al, 2007;Orzaez et al, 2006;Zia et al, 2011), by injecting A. tumefaciens culture directly into the plant tissue or organs; vacuum infiltration (Nanasato et al, 2013;Bratic et al, 2007), by a vacuum to force A. tumefaciens get in into plant cells;…”

Section: Molecular Analysis Of Transformant Candidates Of Tomato Plantsmentioning

confidence: 99%

PRICK AND SOAK Agroacterium tumefaciens-MEDIATED IN PLANTA TRANSFORMATION IN TOMATO (Lycopersicon esculentum Mill.)

Sanjaya

Dwiyani

Wirawan

et al. 2018

Int. J. Biosci. Biotech.

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One of the modern plant breedings through genetic engineering is Agrobacterium tumefaciens-mediated transformation. Agrobacterium tumefaciens-mediated transformation can be performed in vitro or in planta. In planta transformation arises from the weaknesses of the in vitro method such as need high hygiene standard, professional tissue culture experts, and more time to prepare explants and somaclonal variation. In planta transformation is a method to transfer the gene to the plant genome without any tissue culture stages. The aims of this research were to know the possibility of the prick and soak in planta method with the target of tomato seeds and to know the most suitable inoculation time for tomato seeds transformation by prick and soak method the transformation is done by pricking the seeds and soaking them in the A. tumefaciens suspension. The treatments in this study were 1 and 2 days inoculation time to test the efficacy of prick and soak in planta transformation method. Tomato seeds were pricked with a needle on the center once, and then soaked in A. tumefaciens strain LB4404 suspension carrying pKYS-SoSPS1 plasmid with Neomycin Phosphotransferase (NPTII) and Saccharum officinarum Sucrose Phosphate synthase (SoSPS1) genes. Visualization of tomato’s DNA samples after PCR showed that 1-day inoculation sample was positively integrated with NPTII gene and negative in the 2 days inoculation treatment.

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“…Some improvements in the protocol have been lately reported for melon ( Bezirganoglu et al, 2014 ; Zhang et al, 2014 ), cucumber ( Wang et al, 2015 ), Cucurbita spp. ( Nanasato et al, 2013 ), and watermelon ( Liu et al, 2016 ). In the latter species, an efficient regeneration procedure via SE from embryogenic calli derived from leaf explants has been also recently published ( Vinoth and Ravindhran, 2015 ).…”

Section: Regeneration and Transformationmentioning

confidence: 99%

Genetic Transformation and Genomic Resources for Next-Generation Precise Genome Engineering in Vegetable Crops

2017

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In the frame of modern agriculture facing the predicted increase of population and general environmental changes, the securement of high quality food remains a major challenge to deal with. Vegetable crops include a large number of species, characterized by multiple geographical origins, large genetic variability and diverse reproductive features. Due to their nutritional value, they have an important place in human diet. In recent years, many crop genomes have been sequenced permitting the identification of genes and superior alleles associated with desirable traits. Furthermore, innovative biotechnological approaches allow to take a step forward towards the development of new improved cultivars harboring precise genome modifications. Sequence-based knowledge coupled with advanced biotechnologies is supporting the widespread application of new plant breeding techniques to enhance the success in modification and transfer of useful alleles into target varieties. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 system, zinc-finger nucleases, and transcription activator-like effector nucleases represent the main methods available for plant genome engineering through targeted modifications. Such technologies, however, require efficient transformation protocols as well as extensive genomic resources and accurate knowledge before they can be efficiently exploited in practical breeding programs. In this review, we revise the state of the art in relation to availability of such scientific and technological resources in various groups of vegetables, describe genome editing results obtained so far and discuss the implications for future applications.

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Improving the transformation efficiency of Cucurbita species: factors and strategy for practical application

Cited by 17 publications

References 36 publications

Efficient genetic transformation of Jatropha curcas L. by means of vacuum infiltration combined with filter-paper wicks

Efficient genetic transformation of Jatropha curcas L. by means of vacuum infiltration combined with filter-paper wicks

PRICK AND SOAK Agroacterium tumefaciens-MEDIATED IN PLANTA TRANSFORMATION IN TOMATO (Lycopersicon esculentum Mill.)

Genetic Transformation and Genomic Resources for Next-Generation Precise Genome Engineering in Vegetable Crops

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