Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes

Yanal A. Alkuddsi; Shreekanth S. Patil; S. M. Manjula; K. J. Pranesh; B. C. Patil

doi:doi:10.11648/j.ajls.20140204.11

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Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes

Yanal A. Alkuddsi, Shreekanth S. Patil, S. M. Manjula, K. J. Pranesh, B. C. Patil

Published in American Journal of Life Sciences (Volume 2, Issue 4)

Received: 13 July 2014 Accepted: 22 July 2014 Published: 10 August 2014

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Abstract

Cotton breeding for insect resistance has been limited by a lack of sufficient genetic variation in the existing germplasms. Therefore, genetic engineering provides the possibility of creating varieties carrying new properties coming even from heterologous source. Exogenous pesticidal transgenes can be introduced into plants. Agrobacterium mediated plant transformation offers advantages like reducing copy number of the transgene and little co-suppression. Inter specific hybrids are known to be more susceptible to biotic stress. It is hence important to develop Bt version for inter specific hybrid. Presently, the Bt gene commercialized are owned by private sector. It is necessary to develop public sector’s Bt event and commercialize them. UAS Dharwad is involved in developing public sector Bt cotton genotypes. One variety, RCR4 (Gossypium hirsutum, L.) was used in the present investigation. Cry1Ac-Cry1Ec genes are to control Helicoverpa armigera and Spodoptera litura. The seedlings in pots were co-cultivated with solid Agrobacterium culture after cutting the meristematic tip with sharp knife. The number of seedlings co-cultivated, number of seedlings established and the number of seedlings showing transformed status are presented in this study. PCR was performed to confirm the presence of the transgene in the plants that were selected to be advanced further. The results showed that non of plants had trangenes Cry1Ac- Cry1Ec as detected through PCR amplification. In planta genetic transformation was carried out and the plants were tested in T0 generation by means of PCR amplification for the genes Cry1Ac- Cry1Ec. The results obtained were not amplified the Cry1Ac- Cry1Ec. Hence the transformation of the genes was not up to mark and the plants of T1 generation are also not confirmed.

Published in	American Journal of Life Sciences (Volume 2, Issue 4)
DOI	10.11648/j.ajls.20140204.11
Page(s)	190-199
Creative Commons	This is an Open Access article, 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 or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2014. Published by Science Publishing Group

Keywords

Genetic Transformation, Agrobacterium Tumefacience, G. Hirsutum, Cry1Ac-Cry1Ec Genes

References

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Yanal A. Alkuddsi, Shreekanth S. Patil, S. M. Manjula, K. J. Pranesh, B. C. Patil. (2014). Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes. American Journal of Life Sciences, 2(4), 190-199. https://doi.org/10.11648/j.ajls.20140204.11

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ACS Style

Yanal A. Alkuddsi; Shreekanth S. Patil; S. M. Manjula; K. J. Pranesh; B. C. Patil. Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes. Am. J. Life Sci. 2014, 2(4), 190-199. doi: 10.11648/j.ajls.20140204.11

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AMA Style

Yanal A. Alkuddsi, Shreekanth S. Patil, S. M. Manjula, K. J. Pranesh, B. C. Patil. Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes. Am J Life Sci. 2014;2(4):190-199. doi: 10.11648/j.ajls.20140204.11

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@article{10.11648/j.ajls.20140204.11,
  author = {Yanal A. Alkuddsi and Shreekanth S. Patil and S. M. Manjula and K. J. Pranesh and B. C. Patil},
  title = {Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes},
  journal = {American Journal of Life Sciences},
  volume = {2},
  number = {4},
  pages = {190-199},
  doi = {10.11648/j.ajls.20140204.11},
  url = {https://doi.org/10.11648/j.ajls.20140204.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajls.20140204.11},
  abstract = {Cotton breeding for insect resistance has been limited by a lack of sufficient genetic variation in the existing germplasms. Therefore, genetic engineering provides the possibility of creating varieties carrying new properties coming even from heterologous source. Exogenous pesticidal transgenes can be introduced into plants. Agrobacterium mediated plant transformation offers advantages like reducing copy number of the transgene and little co-suppression. Inter specific hybrids are known to be more susceptible to biotic stress. It is hence important to develop Bt version for inter specific hybrid. Presently, the Bt gene commercialized are owned by private sector. It is necessary to develop public sector’s Bt event and commercialize them. UAS Dharwad is involved in developing public sector Bt cotton genotypes. One variety, RCR4 (Gossypium hirsutum, L.) was used in the present investigation. Cry1Ac-Cry1Ec genes are to control Helicoverpa armigera and Spodoptera litura. The seedlings in pots were co-cultivated with solid Agrobacterium culture after cutting the meristematic tip with sharp knife. The number of seedlings co-cultivated, number of seedlings established and the number of seedlings showing transformed status are presented in this study. PCR was performed to confirm the presence of the transgene in the plants that were selected to be advanced further. The results showed that non of plants had trangenes Cry1Ac- Cry1Ec as detected through PCR amplification. In planta genetic transformation was carried out and the plants were tested in T0 generation by means of PCR amplification for the genes Cry1Ac- Cry1Ec. The results obtained were not amplified the Cry1Ac- Cry1Ec. Hence the transformation of the genes was not up to mark and the plants of T1 generation are also not confirmed.},
 year = {2014}
}

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TY  - JOUR
T1  - Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes
AU  - Yanal A. Alkuddsi
AU  - Shreekanth S. Patil
AU  - S. M. Manjula
AU  - K. J. Pranesh
AU  - B. C. Patil
Y1  - 2014/08/10
PY  - 2014
N1  - https://doi.org/10.11648/j.ajls.20140204.11
DO  - 10.11648/j.ajls.20140204.11
T2  - American Journal of Life Sciences
JF  - American Journal of Life Sciences
JO  - American Journal of Life Sciences
SP  - 190
EP  - 199
PB  - Science Publishing Group
SN  - 2328-5737
UR  - https://doi.org/10.11648/j.ajls.20140204.11
AB  - Cotton breeding for insect resistance has been limited by a lack of sufficient genetic variation in the existing germplasms. Therefore, genetic engineering provides the possibility of creating varieties carrying new properties coming even from heterologous source. Exogenous pesticidal transgenes can be introduced into plants. Agrobacterium mediated plant transformation offers advantages like reducing copy number of the transgene and little co-suppression. Inter specific hybrids are known to be more susceptible to biotic stress. It is hence important to develop Bt version for inter specific hybrid. Presently, the Bt gene commercialized are owned by private sector. It is necessary to develop public sector’s Bt event and commercialize them. UAS Dharwad is involved in developing public sector Bt cotton genotypes. One variety, RCR4 (Gossypium hirsutum, L.) was used in the present investigation. Cry1Ac-Cry1Ec genes are to control Helicoverpa armigera and Spodoptera litura. The seedlings in pots were co-cultivated with solid Agrobacterium culture after cutting the meristematic tip with sharp knife. The number of seedlings co-cultivated, number of seedlings established and the number of seedlings showing transformed status are presented in this study. PCR was performed to confirm the presence of the transgene in the plants that were selected to be advanced further. The results showed that non of plants had trangenes Cry1Ac- Cry1Ec as detected through PCR amplification. In planta genetic transformation was carried out and the plants were tested in T0 generation by means of PCR amplification for the genes Cry1Ac- Cry1Ec. The results obtained were not amplified the Cry1Ac- Cry1Ec. Hence the transformation of the genes was not up to mark and the plants of T1 generation are also not confirmed.
VL  - 2
IS  - 4
ER  -

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Author Information

Yanal A. Alkuddsi

Agricultural Research Station, University of Agricultural Sciences, Dharwad- 580005, Karnataka, India
Shreekanth S. Patil

Agricultural Research Station, University of Agricultural Sciences, Dharwad- 580005, Karnataka, India
S. M. Manjula

Agricultural Research Station, University of Agricultural Sciences, Dharwad- 580005, Karnataka, India
K. J. Pranesh

Agricultural Research Station, University of Agricultural Sciences, Dharwad- 580005, Karnataka, India
B. C. Patil

Agricultural Research Station, University of Agricultural Sciences, Dharwad- 580005, Karnataka, India

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APA Style

Yanal A. Alkuddsi, Shreekanth S. Patil, S. M. Manjula, K. J. Pranesh, B. C. Patil. (2014). Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes. American Journal of Life Sciences, 2(4), 190-199. https://doi.org/10.11648/j.ajls.20140204.11

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ACS Style

Yanal A. Alkuddsi; Shreekanth S. Patil; S. M. Manjula; K. J. Pranesh; B. C. Patil. Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes. Am. J. Life Sci. 2014, 2(4), 190-199. doi: 10.11648/j.ajls.20140204.11

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AMA Style

Yanal A. Alkuddsi, Shreekanth S. Patil, S. M. Manjula, K. J. Pranesh, B. C. Patil. Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes. Am J Life Sci. 2014;2(4):190-199. doi: 10.11648/j.ajls.20140204.11

Copy | Download

@article{10.11648/j.ajls.20140204.11,
  author = {Yanal A. Alkuddsi and Shreekanth S. Patil and S. M. Manjula and K. J. Pranesh and B. C. Patil},
  title = {Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes},
  journal = {American Journal of Life Sciences},
  volume = {2},
  number = {4},
  pages = {190-199},
  doi = {10.11648/j.ajls.20140204.11},
  url = {https://doi.org/10.11648/j.ajls.20140204.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajls.20140204.11},
  abstract = {Cotton breeding for insect resistance has been limited by a lack of sufficient genetic variation in the existing germplasms. Therefore, genetic engineering provides the possibility of creating varieties carrying new properties coming even from heterologous source. Exogenous pesticidal transgenes can be introduced into plants. Agrobacterium mediated plant transformation offers advantages like reducing copy number of the transgene and little co-suppression. Inter specific hybrids are known to be more susceptible to biotic stress. It is hence important to develop Bt version for inter specific hybrid. Presently, the Bt gene commercialized are owned by private sector. It is necessary to develop public sector’s Bt event and commercialize them. UAS Dharwad is involved in developing public sector Bt cotton genotypes. One variety, RCR4 (Gossypium hirsutum, L.) was used in the present investigation. Cry1Ac-Cry1Ec genes are to control Helicoverpa armigera and Spodoptera litura. The seedlings in pots were co-cultivated with solid Agrobacterium culture after cutting the meristematic tip with sharp knife. The number of seedlings co-cultivated, number of seedlings established and the number of seedlings showing transformed status are presented in this study. PCR was performed to confirm the presence of the transgene in the plants that were selected to be advanced further. The results showed that non of plants had trangenes Cry1Ac- Cry1Ec as detected through PCR amplification. In planta genetic transformation was carried out and the plants were tested in T0 generation by means of PCR amplification for the genes Cry1Ac- Cry1Ec. The results obtained were not amplified the Cry1Ac- Cry1Ec. Hence the transformation of the genes was not up to mark and the plants of T1 generation are also not confirmed.},
 year = {2014}
}

Copy | Download

TY  - JOUR
T1  - Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes
AU  - Yanal A. Alkuddsi
AU  - Shreekanth S. Patil
AU  - S. M. Manjula
AU  - K. J. Pranesh
AU  - B. C. Patil
Y1  - 2014/08/10
PY  - 2014
N1  - https://doi.org/10.11648/j.ajls.20140204.11
DO  - 10.11648/j.ajls.20140204.11
T2  - American Journal of Life Sciences
JF  - American Journal of Life Sciences
JO  - American Journal of Life Sciences
SP  - 190
EP  - 199
PB  - Science Publishing Group
SN  - 2328-5737
UR  - https://doi.org/10.11648/j.ajls.20140204.11
AB  - Cotton breeding for insect resistance has been limited by a lack of sufficient genetic variation in the existing germplasms. Therefore, genetic engineering provides the possibility of creating varieties carrying new properties coming even from heterologous source. Exogenous pesticidal transgenes can be introduced into plants. Agrobacterium mediated plant transformation offers advantages like reducing copy number of the transgene and little co-suppression. Inter specific hybrids are known to be more susceptible to biotic stress. It is hence important to develop Bt version for inter specific hybrid. Presently, the Bt gene commercialized are owned by private sector. It is necessary to develop public sector’s Bt event and commercialize them. UAS Dharwad is involved in developing public sector Bt cotton genotypes. One variety, RCR4 (Gossypium hirsutum, L.) was used in the present investigation. Cry1Ac-Cry1Ec genes are to control Helicoverpa armigera and Spodoptera litura. The seedlings in pots were co-cultivated with solid Agrobacterium culture after cutting the meristematic tip with sharp knife. The number of seedlings co-cultivated, number of seedlings established and the number of seedlings showing transformed status are presented in this study. PCR was performed to confirm the presence of the transgene in the plants that were selected to be advanced further. The results showed that non of plants had trangenes Cry1Ac- Cry1Ec as detected through PCR amplification. In planta genetic transformation was carried out and the plants were tested in T0 generation by means of PCR amplification for the genes Cry1Ac- Cry1Ec. The results obtained were not amplified the Cry1Ac- Cry1Ec. Hence the transformation of the genes was not up to mark and the plants of T1 generation are also not confirmed.
VL  - 2
IS  - 4
ER  -

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