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Herbicide-resistant cotton (Gossypium hirsutum) plants: an alternative way of manual weed removal

BACKGROUND: Cotton yield has been badly affected by different insects and weed competition. In Past Application of multiple chemicals is required to manage insects and weed control was achieved by different conventional means, such as hand weeding, crop rotation and polyculture, because no synthetic...

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Autores principales: Latif, Ayesha, Rao, Abdul Qayyum, Khan, Muhammad Azmat Ullah, Shahid, Naila, Bajwa, Kamran Shehzad, Ashraf, Muhammad Aleem, Abbas, Malik Adil, Azam, Muhammad, Shahid, Ahmad Ali, Nasir, Idrees Ahmad, Husnain, Tayyab
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4574545/
https://www.ncbi.nlm.nih.gov/pubmed/26383095
http://dx.doi.org/10.1186/s13104-015-1397-0
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author Latif, Ayesha
Rao, Abdul Qayyum
Khan, Muhammad Azmat Ullah
Shahid, Naila
Bajwa, Kamran Shehzad
Ashraf, Muhammad Aleem
Abbas, Malik Adil
Azam, Muhammad
Shahid, Ahmad Ali
Nasir, Idrees Ahmad
Husnain, Tayyab
author_facet Latif, Ayesha
Rao, Abdul Qayyum
Khan, Muhammad Azmat Ullah
Shahid, Naila
Bajwa, Kamran Shehzad
Ashraf, Muhammad Aleem
Abbas, Malik Adil
Azam, Muhammad
Shahid, Ahmad Ali
Nasir, Idrees Ahmad
Husnain, Tayyab
author_sort Latif, Ayesha
collection PubMed
description BACKGROUND: Cotton yield has been badly affected by different insects and weed competition. In Past Application of multiple chemicals is required to manage insects and weed control was achieved by different conventional means, such as hand weeding, crop rotation and polyculture, because no synthetic chemicals were available. The control methods shifted towards high input and target-oriented methods after the discovery of synthetic herbicide in the 1930s. To utilise the transgenic approach, cotton plants expressing the codon-optimised CEMB GTGene were produced in the present study. RESULTS: Local cotton variety CEMB-02 containing Cry1Ac and Cry2A in single cassette was transformed by synthetic codon-optimised 5-enolpyruvylshikimate-3-phosphate synthase gene cloned into pCAMBIA 1301 vector under 35S promoter with Agrobacterium tumifaciens. Putative transgenic plants were screened in MS medium containing 120 µmol/L glyphosate. Integration and expression of the gene were evaluated by PCR from genomic DNA and ELISA from protein. A 1.4-kb PCR product for Glyphosate and 167-bp product for Cry2A were obtained by amplification through gene specific primers. Expression level of Glyphosate and Bt proteins in two transgenic lines were recorded to be 0.362, 0.325 µg/g leaf and 0.390, 0.300 µg/g leaf respectively. FISH analysis of transgenic lines demonstrates the presence of one and two copy no. of Cp4 EPSPS transgene respectively. Efficacy of the transgene Cp4 EPSPS was further evaluated by Glyphosate spray (41 %) assay at 1900 ml/acre and insect bioassay which shows 100 %mortality of insect feeding on transgenic lines as compared to control. CONCLUSION: The present study shows that the transgenic lines produced in this study were resistant not only to insects but also equally good against 1900 ml/acre field spray concentration of glyphosate. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13104-015-1397-0) contains supplementary material, which is available to authorized users.
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spelling pubmed-45745452015-09-19 Herbicide-resistant cotton (Gossypium hirsutum) plants: an alternative way of manual weed removal Latif, Ayesha Rao, Abdul Qayyum Khan, Muhammad Azmat Ullah Shahid, Naila Bajwa, Kamran Shehzad Ashraf, Muhammad Aleem Abbas, Malik Adil Azam, Muhammad Shahid, Ahmad Ali Nasir, Idrees Ahmad Husnain, Tayyab BMC Res Notes Research Article BACKGROUND: Cotton yield has been badly affected by different insects and weed competition. In Past Application of multiple chemicals is required to manage insects and weed control was achieved by different conventional means, such as hand weeding, crop rotation and polyculture, because no synthetic chemicals were available. The control methods shifted towards high input and target-oriented methods after the discovery of synthetic herbicide in the 1930s. To utilise the transgenic approach, cotton plants expressing the codon-optimised CEMB GTGene were produced in the present study. RESULTS: Local cotton variety CEMB-02 containing Cry1Ac and Cry2A in single cassette was transformed by synthetic codon-optimised 5-enolpyruvylshikimate-3-phosphate synthase gene cloned into pCAMBIA 1301 vector under 35S promoter with Agrobacterium tumifaciens. Putative transgenic plants were screened in MS medium containing 120 µmol/L glyphosate. Integration and expression of the gene were evaluated by PCR from genomic DNA and ELISA from protein. A 1.4-kb PCR product for Glyphosate and 167-bp product for Cry2A were obtained by amplification through gene specific primers. Expression level of Glyphosate and Bt proteins in two transgenic lines were recorded to be 0.362, 0.325 µg/g leaf and 0.390, 0.300 µg/g leaf respectively. FISH analysis of transgenic lines demonstrates the presence of one and two copy no. of Cp4 EPSPS transgene respectively. Efficacy of the transgene Cp4 EPSPS was further evaluated by Glyphosate spray (41 %) assay at 1900 ml/acre and insect bioassay which shows 100 %mortality of insect feeding on transgenic lines as compared to control. CONCLUSION: The present study shows that the transgenic lines produced in this study were resistant not only to insects but also equally good against 1900 ml/acre field spray concentration of glyphosate. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13104-015-1397-0) contains supplementary material, which is available to authorized users. BioMed Central 2015-09-17 /pmc/articles/PMC4574545/ /pubmed/26383095 http://dx.doi.org/10.1186/s13104-015-1397-0 Text en © Latif et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Latif, Ayesha
Rao, Abdul Qayyum
Khan, Muhammad Azmat Ullah
Shahid, Naila
Bajwa, Kamran Shehzad
Ashraf, Muhammad Aleem
Abbas, Malik Adil
Azam, Muhammad
Shahid, Ahmad Ali
Nasir, Idrees Ahmad
Husnain, Tayyab
Herbicide-resistant cotton (Gossypium hirsutum) plants: an alternative way of manual weed removal
title Herbicide-resistant cotton (Gossypium hirsutum) plants: an alternative way of manual weed removal
title_full Herbicide-resistant cotton (Gossypium hirsutum) plants: an alternative way of manual weed removal
title_fullStr Herbicide-resistant cotton (Gossypium hirsutum) plants: an alternative way of manual weed removal
title_full_unstemmed Herbicide-resistant cotton (Gossypium hirsutum) plants: an alternative way of manual weed removal
title_short Herbicide-resistant cotton (Gossypium hirsutum) plants: an alternative way of manual weed removal
title_sort herbicide-resistant cotton (gossypium hirsutum) plants: an alternative way of manual weed removal
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4574545/
https://www.ncbi.nlm.nih.gov/pubmed/26383095
http://dx.doi.org/10.1186/s13104-015-1397-0
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