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Sudhakar, D.; Duc, Le Tan; Bong, Bui Ba; Tinjuangjun, Porntip; Maqbool, Shahina; Valdez, Marta; Jefferson, Richard A.; Christou, Paul

doi:10.1023/a:1008870012568

Cited by 54 publications

(9 citation statements)

References 12 publications

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“…The control and fusion protein cassettes were isolated from the intermediate vectors by using EheI and HindIII and directionally subcloned in pAL76 (a ubiquitin promoter-based transformation vector) previously digested with SmaI and HindIII. Transformation of maize and rice embryogenic callus was carried out by particle bombardment as described (21,22). In each transformation procedure, the maize callus was cobombarded with a plasmid carrying the bar selectable marker for phosphinothricin resistance, and the rice callus was cobombarded with the hpt selectable marker for hygromycin resistance.…”

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confidence: 99%

An alternative strategy for sustainable pest resistance in genetically enhanced crops

Mehlo

Gahakwa²,

Nghia³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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104

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Bacillus thuringiensis (Bt) crystal protein genes encode insecticidal ␦-endotoxins that are widely used for the development of insectresistant crops. In this article, we describe an alternative transgenic strategy that has the potential to generate broader and more sustainable levels of resistance against insect pests. Our strategy involves engineering plants with a fusion protein combining the ␦-endotoxin Cry1Ac with the galactose-binding domain of the nontoxic ricin B-chain (RB). This fusion, designated BtRB, provides the toxin with additional, binding domains, thus increasing the potential number of interactions at the molecular level in target insects. Transgenic rice and maize plants engineered to express the fusion protein were significantly more toxic in insect bioassays than those containing the Bt gene alone. They were also resistant to a wider range of insects, including important pests that are not normally susceptible to Bt toxins. The potential impact of fusion genes such as BtRB in terms of crop improvement, resistance sustainability, and biosafety is discussed.Bt genes ͉ transgenic plants ͉ transgenic maize ͉ transgenic rice T ransgenic plants expressing Bacillus thuringiensis (Bt) toxins have been used successfully to provide resistance against selected insect pests for several years. Indeed, insect resistance is the second most widely used trait in transgenic crops (after herbicide tolerance) in world agriculture (1-5). One potential problem with Bt genes is that Bt insecticides are very widely used, with up to 90% of microbiological insect control products based on topically applied Bt toxins. For this reason, there is concern that insects might evolve resistance to Bt toxins (6, 7). The diamondback moth (Plutella xylostella) has evolved resistance in some open field populations in response to repeated exposure to foliar sprays containing Bt proteins (8), whereas laboratory selection experiments with other insect pests have shown that recessive mutant alleles can confer resistance to multiple Bt toxins (7-10). However, note that the evolution of resistance in insects against transgenic plants expressing Bt toxins has yet to be seen in the field.Recent strategies to address potential limitations in conventional transgenic insect pest control include the stacking or pyramiding of multiple transgenes in the same transgenic plant (11) and the use of hybrid toxins (e.g., fusions between a synthetic truncated Cry1Ba and domain II from Cry1Ia; ref. 12). We have devised an alternative strategy in which the Bt toxin Cry1Ac is fused to the nontoxic ricin B-chain (RB). The recognition of toxin-binding sites in the insect midgut is an important factor determining the spectrum of Bt toxin activity and the severity of toxemia (13). Several groups investigating the mechanism of toxin recognition have identified N-acetyl galactosamine residues as an important component of Bt toxinbinding receptors (14-16). Therefore, we selected the ricin B subunit as a fusion partner for the Bt toxin because RB is a galactose...

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confidence: 99%

An alternative strategy for sustainable pest resistance in genetically enhanced crops

Mehlo

Gahakwa²,

Nghia³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

104

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“…Since hygromycin was used for the selection of transformants in monocotyledonous plant species, such as barley [11], rice [12,13], and sugarcane [14], we preliminary investigated the efficacy of HPT gene as a selection marker for transformed bamboo cells [9]. In order to improve transformation technology for engineering/understanding complex biosynthetic pathway(s) of bamboo plants, we described here an illustration of technical details for generating stable transgenic bamboo suspension cells using the particle bombardment method with pSTARA vector harboring the mutated ALS gene derived from rice.…”

Section: Resultsmentioning

confidence: 99%

The Mutated Acetolactate Synthase Gene from Rice as a Non-Antibiotic Selection Marker for Transformation of Bamboo Cells

Ogita

Kikuchi²,

Nomura³

et al. 2012

AJPS

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Previously, we developed a particle bombardment-mediated transformation protocol in Phyllostachys nigra bamboo by expressing hygromycin phosphotransferase gene (HPT) and neomycin phosphotransferase II gene (NPT II). Although these marker genes could introduce to several tissue cultured organs (e.g. leaves, buds, and calli) of Phyllostachs bamboo species, some organs showed a high susceptibility and/or a low selectivity to hygromycin and kanamycin. In this report, therefore, we describe advantages and technical details for generating stable transgenic bamboo cells using the particle bombardment method with the mutated-acetolactate synthase gene (mALS) from rice (W548L/S627IOsALS) as a non-antibiotic selection marker. A facile and efficient transformation was achieved with the mALS gene and enhanced fluorescent protein gene (mCherry). Approximately 490 and 1400 mCherry-expressing cells/dish/shot in average were observed in both P. bambusoides and P. nigra under fluorescent stereo-microscope. Stable transgenic bamboo cell lines were generated in a selection medium supplemented with 0.1 μM of bispyribac-sodium (BS) as ALS inhibitor. The integration of mALS gene was identified by in vivo ALS enzyme assay and a PCR-restriction fragment length polymerphism (RFLP) based detection procedures

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“…Embryogenic calli obtained from each induction medium were transferred to different regeneration treatments. Basal regeneration medium was similar to that described by Sudhakar et al [ 31 ], and was constituted by MS mineral salts and vitamins, 20 g L − 1 sucrose and 0.3 g L − 1 hydrolyzed casein. Treatments consisting of supplementing the basal medium with variations of growth regulators, and the control was supplemented with 0.5 mg L − 1 α-Naphthaleneacetic Acid (NAA) + 3 mg L − 1 6-BA, while the second treatment was supplemented with 0.5 mg L − 1 NAA + 0.5 mg L − 1 6-BA and the third treatment was supplemented with 0.5 mg L − 1 NAA + 1.5 mg L − 1 Kinetin (KIN) and a fourth treatment consisted of supplementing basal medium with 0.5 mg L − 1 NAA + 0.5 mg L − 1 TDZ.…”

Section: Methodsmentioning

confidence: 99%

“…These response variables were analyzed with a generalized linear model with a Poisson distribution and logit link function. Post hoc analysis consisted of an Honest Significant Difference test on IBM SPSS version 27 [ 31 ] and differences between each combination of factors were recognized.…”

Section: Methodsmentioning

confidence: 99%

“…The survival rate was recorded after calli were transferred to the regeneration medium composed by basal regeneration medium supplemented with 0.5 mg L −1 NAA + 3 mg L −1 6-BA, selected after previous experiments were analyzed. Culture conditions were the same indicated above and after 4 weeks lethal doses were calculated using probit analysis on IBM SPSS version 27 [ 31 ] based on calli death rates.…”

Section: Methodsmentioning

confidence: 99%

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A Temporary Immersion System Improves Regeneration of In Vitro Irradiated Recalcitrant Indica Rice (Oryza sativa L.) Embryogenic Calli

Hernández-Soto

Pérez

Fait-Zúñiga

et al. 2022

Plants

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The development of gamma ray-mutated rice lines is a solution for introducing genetic variability in indica rice varieties already being used by farmers. In vitro gamma ray (60Co) mutagenesis reduces chimeras and allows for a faster selection of desirable traits but requires the optimization of the laboratory procedure. The objectives of the present work were sequencing of matK and rbcL, the in vitro establishment of recalcitrant rice embryogenic calli, the determination of their sensitivity to gamma radiation, and optimization of the generation procedure. All sequenced genes matched perfectly with previously reported matK and rbcL O. sativa genes. Embryogenic calli induction improved using MS medium containing 2 mg L−1 2,4-D, and regeneration was achieved with MS medium with 3 mg L−1 BA and 0.5 mg L−1 NAA. The optimized radiation condition was 60 Gy, (LD20 = 64 Gy) with 83% regeneration. An immersion system (RITA®, Saint-Mathieu-de-Tréviers, France) of either 60 or 120 s every 8 h allowed systematic and homogeneous total regeneration of the recalcitrant line. Other well-known recalcitrant cultivars, CR1821 and CR1113, also had improved regeneration in the immersion system. To our knowledge, this is the first study reporting the use of an immersion system to allow for the regeneration of gamma-ray mutants from recalcitrant indica rice materials.

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Cited by 54 publications

References 12 publications

An alternative strategy for sustainable pest resistance in genetically enhanced crops

An alternative strategy for sustainable pest resistance in genetically enhanced crops

The Mutated Acetolactate Synthase Gene from Rice as a Non-Antibiotic Selection Marker for Transformation of Bamboo Cells

A Temporary Immersion System Improves Regeneration of In Vitro Irradiated Recalcitrant Indica Rice (Oryza sativa L.) Embryogenic Calli

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