Genetic transformation of sweet potato by particle bombardment

Prakash, Channapatna S.; Varadarajan, Usha

doi:10.1007/bf00235252

Cited by 46 publications

(15 citation statements)

References 13 publications

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“…Control explants that were not bombarded or those that were bombarded with microcarriers not coated with DNA did not exhibit any uid A expressing cells. Similar uid A expression patterns were observed for cowpea and sweet potato previously (Kononowicz et al, 1995;Prakash and Varadarajan, 1992). Based on the transient uid A expression, particle size, target distance, helium pressure, number of bombardments and vacuum significantly affected transient expression of reporter genes.…”

Section: Optimization Of Conditions For Dna Deliverysupporting

confidence: 81%

Stable gene transformation in cowpea (Vigna unguiculata L. walp.) using particle gun method

Ikea¹,

Ingelbrecht²,

Uwaifo³

et al. 2003

Afr. J. Biotechnol.

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We investigated the possibility of transforming and obtaining transgenic cowpea (Vigna unguiculata L Walp) plants using the particle bombardment process. Meristematic explants that could give rise to whole fertile plants were used in transformation experiments with reporter and selectable marker genes driven by a 35S CaMV promoter. Conditions for optimal delivery of DNA to explants were established based on transient gus expression assays two days after bombardment. The size of microcarriers, microflight distance and helium pressure significantly affected transient expression of reporter genes. A total of 1692 explants were bombarded with DNA-coated particles and placed on 3 mg/l bialaphos selective medium. Only 12 regenerated shoots produced seeds eventually, and all were Gus negative even though

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Section: Optimization Of Conditions For Dna Deliverysupporting

confidence: 81%

Stable gene transformation in cowpea (Vigna unguiculata L. walp.) using particle gun method

Ikea¹,

Ingelbrecht²,

Uwaifo³

et al. 2003

Afr. J. Biotechnol.

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show abstract

“…To compare the effects of mannose on growth of sweetpotato callus with that of negative selection system used in several sweetpotato transformation studies (Dhir et al, 1998;Gama et al, 1996;Morán et al, 1998;Newell et al, 1995;Otani et al, 1993;Otani et al, 1998;Prakash and Varadarajan, 1992;Santa-Maria et al, 2011;Song et al, 2004) and determine the effectiveness and optimum concentration for use in the selection of transformed cells, effects of kanamycin and hygromycin on sweetpotato explants were evaluated. Sweetpotato stem explants were transferred onto supplemented with CIM 30 g/l sucrose.…”

Section: Sensitivity Of To Antibiotics Sweetpotato Callusmentioning

confidence: 99%

Evaluation of the Phosphomannose Isomerase-Based Selection System for Genetic Transformation of Sweetpotato

et al. 2015

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Article InformationIn response to increased public concern on antibiotic or herbicide resistance genes usage in genetically modified plants, mannose was evaluated as selectable agent for the genetic transformation of sweetpotato. Nontransformed sweetpotato stem explants of cv. KSP36 were cultured on media containing various combinations and concentrations sucrose and mannose ranging from 0 to 30 g/l. Likewise, efficacy of hygromycin and kanamycin as selection agents for transformation of sweetpotato was evaluated on media containing varying concentrations of antibiotics ranging 0 to 100 mg/l. Selection agent effectiveness was determined by detecting the minimal concentration of the selection agent required to fully inhibit sweetpotato calli growth. Hygromycin was the most effective selection agent as it inhibited cell growth at concentrations above 20 mg/l. Kanamycin was moderately effective as it inhibited cell growth at 60 mg/l. Sweetpotato calli were able to grow and even produce embryos even when mannose was the only source of carbohydrates.

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“…Although sweetpotato has already been transformed (Prakash and Varadarajan, 1992;Newell et al, 1995;Okada et al, 1995Okada et al, , 2001Gama et al, 1996;Min et al, 1998;Mor~n et al, 1998;Otani et al, 1998;Kimura et al, 2001;Wakita et al, 2001 ;Yamaguchi et al, 2004), a more efficient system is needed. The choice of selectable marker and selection agent is critical because of currently low transformation frequencies.…”

mentioning

confidence: 96%

Selectionof nptll transgenic sweetpotato plants using G418 and paromomycin

et al. 2007

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We have used two aminoglycosides, G418 and paromomycin, to develop a reliable selection system for nptll transgenic sweetpotato (Ipomoea batatas (L.) Lain.). Embryogenic calli derived from shoot apical meristems were bombarded with gold particles coated with pCAMBIA2301, which contained the nptll and gusA genes. When compared on a kill curve that was based on calli proliferation and cell viability, G4~8-selection proved to be more efficient and had fewer escapes than kanamycin. These bombarded explants were then selected on G418-containing media. The total time required from bombardment to plant establishment in soil was seven to nine months. Multiple copies of the transgene were integrated into the sweetpotato genome. Northern analysis confirmed transgene expression in the regenerated plants, and a paromomycin assay demonstrated that the npUl gene was functionally expressed in transformed sweetpotato. These molecular analyses and assays all showed that selection with G418 and paromomycin is reliable. So far, we have produced 69 transgenic events with this system, at a transformation frequency of approx. 1.1%. That efficiency is based on the number of transgenic plants obtained and the amount of calli bombarded. Thus, this selection method that combines G41a with paromomycin is now available for selecting npUl transgenic sweetpolato.

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Genetic transformation of sweet potato by particle bombardment

Cited by 46 publications

References 13 publications

Stable gene transformation in cowpea (Vigna unguiculata L. walp.) using particle gun method

Stable gene transformation in cowpea (Vigna unguiculata L. walp.) using particle gun method

Evaluation of the Phosphomannose Isomerase-Based Selection System for Genetic Transformation of Sweetpotato

Selectionof nptll transgenic sweetpotato plants using G418 and paromomycin

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