Expression of cry1Aa gene in cabbage imparts resistance against diamondback moth (Plutella xylostella)

Abstract

Brassica oleracea cv. Pride of India is one of the most promising vegetable cultivars commercially grown as cash crop in Himachal Pradesh, India. However, its overall production is severely hampered by diamondback moth (Plutella xylostella), a notorious pest. To avoid yield losses caused by this pest, cryIAa gene was introduced into cabbage cv. Pride of India using Agrobacterium-mediated transformation method. In an attempt to maximize the transformation frequency, critical determinant factors such as explant type, pre-incubation and co-cultivation period, and acetosyringone effect were successfully optimized. The highest transformation frequency (4.67% and 14.50%) in cotyledon and hypocotyl explant was achieved with a pre-incubation period of 72 h and co-cultivation period of 48 h. Furthermore, transformation frequency was enhanced in cotyledon (18.66%) and hypocotyl (32.00%) explants, when selective regeneration medium was fortified with 100 µM acetosyringone, respectively. The transgene (cryIAa) integration and copy number were confirmed using PCR and Southern blotting. Reverse transcriptase PCR and quantitative real-time PCR analyses were performed that proved transcriptional expression of cryIAa gene in PCR-positive transgenic events. Transgenic cabbage-fed diamondback moth larvae showed significantly higher mortality, thereby proving transgene effectiveness against insect pest control.

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Acknowledgements

Authors are grateful to Professor and Head, Department of Vegetable Science, Dr. Y. S. Parmar University of Horticulture and Forestry, Solan, for providing the plant material of cabbage cv. Pride of India. We are also highly thankful to the Project Director, ICAR-National Institute of Plant Biotechnology, New Delhi, for providing Agrobacterium strain and for molecular analysis work.

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DKS and GG conceived the research work and designed the experiments. GG, GA and PK executed the laboratory/field experiments and collected data. GG, PK and DKS analyzed and interpreted the data. GG, PK, DKS and AKT were involved in the preparation of the manuscript.

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Correspondence to Ajay Kumar Thakur.

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The authors declare that they have no conflict of interest.

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Gambhir, G., Kumar, P., Aggarwal, G. et al. Expression of cry1Aa gene in cabbage imparts resistance against diamondback moth (Plutella xylostella). BIOLOGIA FUTURA (2020). https://doi.org/10.1007/s42977-020-00014-8

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Keywords

  • Genetic transformation
  • Cotyledon
  • Hypocotyl
  • Insect bioassay
  • cry1Aa gene
  • Insect resistance