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Comparative performance of modified full-length and truncated Bacillus thuringiensis-cry1Ac genes in transgenic tomato

BACKGROUND: Bt-cry1Ac gene has been reputedly effective against Helicoverpa armigera a notorious lepidopteran pest. Reports on the expression of full-length and truncated cry1Ac genes in plants for effective resistance against Helicoverpa sp. have been documented however, their performance is still...

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Autores principales: Koul, Bhupendra, Yadav, Reena, Sanyal, Indraneel, Amla, Devindra Vijay
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4422829/
https://www.ncbi.nlm.nih.gov/pubmed/25977893
http://dx.doi.org/10.1186/s40064-015-0991-x
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author Koul, Bhupendra
Yadav, Reena
Sanyal, Indraneel
Amla, Devindra Vijay
author_facet Koul, Bhupendra
Yadav, Reena
Sanyal, Indraneel
Amla, Devindra Vijay
author_sort Koul, Bhupendra
collection PubMed
description BACKGROUND: Bt-cry1Ac gene has been reputedly effective against Helicoverpa armigera a notorious lepidopteran pest. Reports on the expression of full-length and truncated cry1Ac genes in plants for effective resistance against Helicoverpa sp. have been documented however, their performance is still ambiguous. Moreover, the question remains to be addressed that truncation of 3′ end of the native gene was documented and suggested for active insecticidal toxin production while the most successful transgenic event(s) of commercialized-cotton are based on full-length of the cry gene. Therefore, we performed a comparative study on the efficacy of the two versions of cry1Ac genes (full-length: 3,510 bp and truncated: 1,845 bp) in T(0) and T(1) transgenic tomato plants and analyzed the extent of protection against H. armigera and also compared the results with our previous findings related to a successful transgenic tomato line Ab25E, expressing cry1Ab gene. The integration of cry1Ac gene(s) in T(0) transgenic plants and its inheritance in T(1) progeny was observed by PCR, RT-PCR and Southern blot hybridization analysis while, the toxin integrity, expression and toxicity was monitored by Western immunoassay, DAS-ELISA and insect bioassay respectively. RESULTS: An average transformation frequency and Bt-Cry protein content of 16.93 ± 2.10 and 0.0020–0.0128% of total soluble protein (TSP) was obtained with pRD400 vector (Trcry1Ac) while, a much lower value of 9.30 ± 2.041 and 0.0001 ― 0.0026% of TSP was observed with pNBRI-1 vector (Flcry1Ac), respectively. The promising Trcry1Ac T(0) transgenic plants and their T(1) progeny gave full protection from H. armigera. Although Flcry1Ac gene showed lower transformation frequency and lower expression, it showed higher toxicity to H. armigera when compared with truncated Trcry1Ac gene. CONCLUSIONS: The full-length cry1Ac gene can be redesigned for higher expression and performance in dicots or a hybrid gene could be designed having a blend of strong receptor binding and stable expression characteristics for enhanced efficacy and toxicity to the susceptible insects. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40064-015-0991-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-44228292015-05-14 Comparative performance of modified full-length and truncated Bacillus thuringiensis-cry1Ac genes in transgenic tomato Koul, Bhupendra Yadav, Reena Sanyal, Indraneel Amla, Devindra Vijay Springerplus Research BACKGROUND: Bt-cry1Ac gene has been reputedly effective against Helicoverpa armigera a notorious lepidopteran pest. Reports on the expression of full-length and truncated cry1Ac genes in plants for effective resistance against Helicoverpa sp. have been documented however, their performance is still ambiguous. Moreover, the question remains to be addressed that truncation of 3′ end of the native gene was documented and suggested for active insecticidal toxin production while the most successful transgenic event(s) of commercialized-cotton are based on full-length of the cry gene. Therefore, we performed a comparative study on the efficacy of the two versions of cry1Ac genes (full-length: 3,510 bp and truncated: 1,845 bp) in T(0) and T(1) transgenic tomato plants and analyzed the extent of protection against H. armigera and also compared the results with our previous findings related to a successful transgenic tomato line Ab25E, expressing cry1Ab gene. The integration of cry1Ac gene(s) in T(0) transgenic plants and its inheritance in T(1) progeny was observed by PCR, RT-PCR and Southern blot hybridization analysis while, the toxin integrity, expression and toxicity was monitored by Western immunoassay, DAS-ELISA and insect bioassay respectively. RESULTS: An average transformation frequency and Bt-Cry protein content of 16.93 ± 2.10 and 0.0020–0.0128% of total soluble protein (TSP) was obtained with pRD400 vector (Trcry1Ac) while, a much lower value of 9.30 ± 2.041 and 0.0001 ― 0.0026% of TSP was observed with pNBRI-1 vector (Flcry1Ac), respectively. The promising Trcry1Ac T(0) transgenic plants and their T(1) progeny gave full protection from H. armigera. Although Flcry1Ac gene showed lower transformation frequency and lower expression, it showed higher toxicity to H. armigera when compared with truncated Trcry1Ac gene. CONCLUSIONS: The full-length cry1Ac gene can be redesigned for higher expression and performance in dicots or a hybrid gene could be designed having a blend of strong receptor binding and stable expression characteristics for enhanced efficacy and toxicity to the susceptible insects. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40064-015-0991-x) contains supplementary material, which is available to authorized users. Springer International Publishing 2015-04-30 /pmc/articles/PMC4422829/ /pubmed/25977893 http://dx.doi.org/10.1186/s40064-015-0991-x Text en © Koul et al.; licensee Springer. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.
spellingShingle Research
Koul, Bhupendra
Yadav, Reena
Sanyal, Indraneel
Amla, Devindra Vijay
Comparative performance of modified full-length and truncated Bacillus thuringiensis-cry1Ac genes in transgenic tomato
title Comparative performance of modified full-length and truncated Bacillus thuringiensis-cry1Ac genes in transgenic tomato
title_full Comparative performance of modified full-length and truncated Bacillus thuringiensis-cry1Ac genes in transgenic tomato
title_fullStr Comparative performance of modified full-length and truncated Bacillus thuringiensis-cry1Ac genes in transgenic tomato
title_full_unstemmed Comparative performance of modified full-length and truncated Bacillus thuringiensis-cry1Ac genes in transgenic tomato
title_short Comparative performance of modified full-length and truncated Bacillus thuringiensis-cry1Ac genes in transgenic tomato
title_sort comparative performance of modified full-length and truncated bacillus thuringiensis-cry1ac genes in transgenic tomato
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4422829/
https://www.ncbi.nlm.nih.gov/pubmed/25977893
http://dx.doi.org/10.1186/s40064-015-0991-x
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