Sweet Potato (Ipomoea batatas L.) Regeneration and Transformation Technology to Provide Weevil (Cylas formicarius) Resistance. Field Trial Results

Garcia, Ricardo Ferreira; Morán, Rolando; Mena, Jesús; Somontes, Danalay; Pimentel, Eulogio; Zaldúa, Zurima; López, Alina; Garcia, M.

doi:10.1016/s0168-7972(00)80017-1

Cited by 9 publications

(6 citation statements)

References 6 publications

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“…Hence, the use of genetically engineered sweet potato plants expressing Cry toxins could control these coleopteran pests, as Bt crops have been shown to effectively control stem borers, ear feeders, and rootworms (10). The first attempt to develop a Bt sweet potato expressing Cry3A did not fully control C. formicarius due to low accumulation of the Cry protein (6,7). Similar results were observed recently when Cry7Aa was expressed in the storage root (11).…”

supporting

confidence: 60%

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Shared Binding Sites for the Bacillus thuringiensis Proteins Cry3Bb, Cry3Ca, and Cry7Aa in the African Sweet Potato Pest Cylas puncticollis (Brentidae)

Hernández‐Martínez

Vera-Velasco

Martínez-Solís

et al. 2014

Appl Environ Microbiol

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bBacillus thuringiensis Cry3Bb, Cry3Ca, and Cry7Aa have been reported to be toxic against larvae of the genus Cylas, which are important pests of sweet potato worldwide and particularly in sub-Saharan Africa. However, relatively little is known about the processing and binding interactions of these coleopteran-specific Cry proteins. The aim of the present study was to determine whether Cry3Bb, Cry3Ca, and Cry7Aa proteins have shared binding sites in Cylas puncticollis to orient the pest resistance strategy by genetic transformation. Interestingly, processing of the 129-kDa Cry7Aa protoxin using commercial trypsin or chymotrypsin rendered two fragments of about 70 kDa and 65 kDa. N-terminal sequencing of the trypsin-activated Cry7Aa fragments revealed that processing occurs at Glu 47 for the 70-kDa form or Ile 88 for the 65-kDa form. Homologous binding assays showed specific binding of the two Cry3 proteins and the 65-kDa Cry7Aa fragment to brush border membrane vesicles (BBMV) from C. puncticollis larvae. The 70-kDa fragment did not bind to BBMV. Heterologous-competition assays showed that Cry3Bb, Cry3Ca, and Cry7Aa (65-kDa fragment) competed for the same binding sites. Hence, our results suggest that pest resistance mediated by the alteration of a shared Cry receptor binding site might render all three Cry toxins ineffective.

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supporting

confidence: 60%

“…The adults lay eggs in the storage root, where larva tunneling leads to rotting, rendering it unsuitable for consumption (4). Current control strategies for these insect pests have not been effective, justifying the need for new control methods such as the use of insecticidal Cry proteins from Bacillus thuringiensis (5)(6)(7).…”

mentioning

confidence: 99%

Shared Binding Sites for the Bacillus thuringiensis Proteins Cry3Bb, Cry3Ca, and Cry7Aa in the African Sweet Potato Pest Cylas puncticollis (Brentidae)

Hernández‐Martínez

Vera-Velasco

Martínez-Solís

et al. 2014

Appl Environ Microbiol

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“…Besides tissue regeneration procedures, methods to produce transgenic sweetpotato plants have also been reported. Stably transformed plants have been obtained by embryogenic regeneration from callus obtained from apical meristems (Gama et al 1996;Otani et al 2001), by somatic embryogenesis regeneration from petioles and stem segments (Cipriani et al 1999;Song et al 2004), and by adventitious shoot regeneration from leaf explants (Morán et al 1999;Garcia et al 2000;Luo et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Rapid shoot regeneration in industrial ‘high starch’ sweetpotato (Ipomoea batatas L.) genotypes

Santa-María

Pecota

Yencho

et al. 2009

Plant Cell Tiss Organ Cult

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“…3) whereas the virC PCR product was absent (Data not shown). These results proved that the transgenic plants were actually transformed with Cry8Db and were not contaminated with Agrobacterium [30]. This result also showed that the use of high concentration of kanamycin at 100 mg/L for selection of putative transformed clones still gave ambiguous results.…”

Section: Molecular Analysis Of Transgenic Plantsmentioning

confidence: 57%

Agrobacterium-Mediated Transformation of Cry8db Gene in Vietnam Sweet Potato Cultivar

Ngoc¹,

Lan²,

Trang³

et al. 2015

JLS

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Sweet potato [Ipomoea batatas (L.) Lam.] is an important food crop in the world as well as in Vietnam. It is well known as a recalcitrant crop for gene transformation and tissue culture because of its genotype dependent in vitro responses. In present study, Agrobacterium-mediated transformation of cry8Db from Bacillus thuringiensis into KB1 sweet potato variety has been studied. The C58cv strain carrying a pBI121 backbone which contained cry8Db delta-endotoxin gene regulated under 35S CaMV promoter, and the selection marker gene, neomycin phosphotransferase (nptII) gene, was subjected for plant transformation. Callus induced from shoot tips and leaf explants were inoculated and cocultured with A. tumefaciens. The selection occurred during callus producing and plant regenerating steps. A total of 201 transgenic putative plant lines were produced, and 21 transgenic lines were positively confirmed by PCR and finalized by Southern blot. Four putative transgenic lines confirming a single copy of the cry8Db gene were transferred into soil pots in greenhouse. Biological activity evaluation for the insecticidal capacity of these transgenic lines under controlled conditions showed that the level of infestation by sweet potato weevils (Cylas formicarius) in untransformed plants was higher than that of transgenic lines.

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Sweet Potato (Ipomoea batatas L.) Regeneration and Transformation Technology to Provide Weevil (Cylas formicarius) Resistance. Field Trial Results

Cited by 9 publications

References 6 publications

Shared Binding Sites for the Bacillus thuringiensis Proteins Cry3Bb, Cry3Ca, and Cry7Aa in the African Sweet Potato Pest Cylas puncticollis (Brentidae)

Shared Binding Sites for the Bacillus thuringiensis Proteins Cry3Bb, Cry3Ca, and Cry7Aa in the African Sweet Potato Pest Cylas puncticollis (Brentidae)

Rapid shoot regeneration in industrial ‘high starch’ sweetpotato (Ipomoea batatas L.) genotypes

Agrobacterium-Mediated Transformation of Cry8db Gene in Vietnam Sweet Potato Cultivar

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