Loading...

Crop Research

Standardization of an efficient protocol for transformation in Indian cauliflower (Brassica oleracea) var. botrytis 


DOI: 10.31830/2454-1761.2022.CR-11088    | Article Id: CR-11088 | Page : 380-385
Citation :- Standardization of an efficient protocol for transformation in Indian cauliflower (Brassica oleracea) var. botrytis. Crop Res. 57: 380-385
SHALINI RAWAT, PRITAM KALIA AND NAVODIT GOEL shalinibiotech05@gmail.com
Address : Amity Institute of Biotechnology, Amity University, Sector 125, Uttar Pradesh, Noida 201313., India
Online Published:

Abstract

   Cauliflower is an important Cole crop grown all over the world for its nutritional benefits. But it is encountered by a serious problem caused by the insect- pest attack. Plutella xylostella is one such insect which causes huge losses. An efficient and simple method for genetic transformation and regeneration of Indian cauliflower, Brassica oleracea var. botrytis has to be developed. Hence, optimization of the plant transformation is a pre- requisite for successful transformation process. Therefore, we worked on this objective at the Indian Agricultural Research Institute, New Delhi, India, and optimized the Agrobacterium-mediated transformation protocol in Indian cauliflower, Pusa Snowball K1 and Pusa Snowball KT25 varieties for developing insect resistance against Plutella xylostella using Bt genes. The Agrobacterium tumefaciens mediated transformation process was utilized for carrying out the transformation and the explant was obtained from the seedlings germinated from the seeds of the respective variety and used for genetic transformation process. The transgene integration and expression were later confirmed by PCR and RT-qPCR, respectively. Moreover, insect bioassay showed that transgenic plants effectively controlled the Diamond Back moth infestation concerning to the control plants indicating the presence of resistance developed against the insect. Hence, this protocol efficiently regenerated transgenic plants that lead to the resistance against biotic stress, specific to the insect Diamond back moth.

Keywords

Agrobacterium cauliflower insect resistance transformation

References

Asande, L. K., Omwoyo, R. O., Oduor, R. O. and Nyaboga, E. N. (2020). A simple and fast Agrobacterium-mediated transformation system for passion fruit KPF4 (Passifora edulis f. edulis × Passifora edulis f. favicarpa). Plant Methods 16 : 1-12.
Bhalla, P. L. and Singh, M. B. (2008). Agrobacterium-mediated transformation of Brassica napus and Brassica oleracea. Nat. Protoc. 3 : 181-89.
Chakrabarty, R., Viswakarma, N., Bhat, S. R., Kirti, P. B., Singh, B, D. and Chopra, V.L. (2002). Agrobacterium-mediated transformation of cauliflower: optimization of protocol and development of  Bt- transgenic cauliflower. J. Biosci. 27 : 495-502.
Deng-xia, Y., Lei, C., Yu-mei, L., Zhuang, M., Yang-yong, Z., Zhi-yuan, F. and Li-mei, Y. (2011). Transformation of Cabbage (Brassica oleracea L. var. capitata) with Bt cry1Ba3 Gene for control of Diamondback Moth. Agric. Sci. China 10 : 1693-700.
Ebrahimzadegan, R. and Maroufi, A. (2022). In-vitro regeneration and Agrobacterium mediated genetic transformation of Dragon's Head plant (Lallemantia iberica). Sci. Rep.12 : 1-12.
Gautam, M. P., Singh, H., Kumar, S., Kumar, V., Singh, G. and Singh, S. N. (2018). Diamondback moth, Plutella xylostella (Linnaeus) (Insecta: Lepidoptera: Plutellidae) a major insect of cabbage in India: A review. J. Entomol. Zool. Stud. 6 : 1394-399.
Gerszberg, A., Konka, K. H. and Kowalczyk, T. (2015). In vitro regeneration of eight cultivars of Brassica oleracea var. capitata. In vitro Cell. Dev. Biol.- Plant 51 : 80-87.
Kowalczyk, T.,  Gerszberg, A., Durańska, P., Biłas, R. and Konka, K. I. (2018). High efficiency transformation of Brassica oleracea var. botrytis plants by Rhizobium rhizogenes. AMB Express  8 : 125-35.
Park, J. R., McFarlane, T., Phipps, R. H. and Ceddia, G. (2011). The role of transgenic crops in sustainable development: A review. Plant Biotechnol. J. 9 : 2–21.
Ramzan, M., Ullah, U. N., Hanif, M., Nadeem, M., Qayyum, M. A., and Javaid, M. (2019). Biology of Diamondback Moth, Plutella xylostella (Lepidoptera: Plutellidae) of Cauliflower under Laboratory Conditions. J. Innov. Sci. 5 : 89-94.
Ravanfar, S. A., Aziz, M. A., Kadir, M. A., Rashid, A. A., and Sirchi, M. H. T. (2009). Plant regeneration of Brassica oleracea subsp. italica (Broccoli) CV Green Marvel as affected by plant growth regulators. Afr. J. Biotechnol. 8 : 2523-528.

Global Footprints