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

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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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References

  • Deng-Xia Y, Lei C, Yu-Mei L, Mu Z, Yang-Yong Z, Zhi-Yuan F, 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(11):1693–1700

    Article  Google Scholar 

  • Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:1–15

    Google Scholar 

  • Gambhir G, Kumar P, Srivastava DK (2017a) High frequency regeneration of plants from cotyledon and hypocotyl cultures in Brassica oleracea cv. Pride of India. Biotechnol Rep 15:107–113

    Article  Google Scholar 

  • Gambhir G, Kumar P, Srivastava DK (2017b) Effect of antibiotic sensitivity on different cultured tissues and its significance in genetic transformation of cabbage Brassica oleracea. Biosci Biotechnol Res Commun 10(4):652–661

    Article  Google Scholar 

  • Gaur A (2015) Studies on Agrobacterium-mediated insect resistance gene [cry1A(a)] transfer in cauliflower (Brassica oleracea L. var. botrytis). Thesis Ph.D., Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni, Solan (H.P.), India

  • Gerszberg A (2018) Tissue culture and genetic transformation of cabbage (Brassica oleracea var. capitata): an overview. Planta 248:1037–1048

    Article  CAS  Google Scholar 

  • Gomez KA, Gomez AA (1984) Statistical procedures for agricultural research. Wiley, New York

    Google Scholar 

  • Hur SH, Min BW (2015) Efficient development of transgenic Cabbage with jasmonic acid carboxyl methyltransferase (JMT) gene based on PMI/mannose selection system. Plant Breed Biotechnol 3(3):226–237

    Article  Google Scholar 

  • Kumar P, Srivastava DK (2016a) Biotechnological advancement in genetic improvement of broccoli (Brassica oleracea L. var. italica), an important vegetable crop: a review. Biotechnol Lett 38(7):1049–1063

    Article  CAS  Google Scholar 

  • Kumar P, Srivastava DK (2016b) Biotechnological application in in vitro plant regeneration studies of Broccoli (Brassica oleracea L. var. italica), an important vegetable crop: a review. Biotechnol Lett 38(4):561–571

    Article  CAS  Google Scholar 

  • Kumar P, Gaur A, Srivastava DK (2017) Agrobacterium- mediated insect resistance Gene (cry1Aa) transfer studies pertaining to antibiotic sensitivity on cultured tissues of broccoli. Intl J Veg Sci 23(6):523–535

    Article  Google Scholar 

  • Kumar P, Gambhir G, Gaur A, Sharma KC, Thakur AK, Srivastava DK (2018) Development of transgenic broccoli with cryIAa gene for resistance against diamondback moth (Plutella xylostella). 3 Biotech 8(7):299

    Article  Google Scholar 

  • Lei C, Mei YL, Nan Z, Mei LY, Mu Z, Yong ZY, Jun ZY, Fang HD, Yuan FZ (2009) Transformation and expressing of Bt gene cry1Ia8 in cabbage. Acta Hortic Sin 36(8):1161–1168

    Google Scholar 

  • Liu H, Guo X, Naeem MS, Liu D, Xu L, Zhang W, Tang G, Zhou W (2011) Transgenic Brassica napus L. lines carrying a two gene construct demonstrate enhanced resistance against Plutella xylostella and Sclerotinia sclerotiorum. Plant Cell Tiss Org Cult 106:143–151

    Article  Google Scholar 

  • Park BJ, Liu J, Kanno A, Kameya T (2005) Genetic improvement of Chinese cabbage for salt and drought tolerance by constitutive expression of a B. napus LEA gene. Plant Sci 169:553–558

    Article  CAS  Google Scholar 

  • Parmar N, Singh KH, Sharma D, Singh L, Kumar P, Nanjundan J, Khan YJ, Chauhan DK, Thakur AK (2017) Genetic engineering strategies for biotic and abiotic stress tolerance and quality enhancement in horticultural crops: a comprehensive review. 3 Biotech 7:239

    Article  Google Scholar 

  • Rafat A, Aziz MA, Rhashid AA, Abdullah SNA, Kamaladini H, Sirchi MHT, Javadi MB (2010) Optimization of Agrobacterium tumefaciens-mediated transformation and shoot regeneration after co-cultivation of cabbage (Brassica oleracea subsp. capitata) cv. KY cross with At HP101 gene. Sci Hortic 124:1–8

    Article  CAS  Google Scholar 

  • Song GQ, Walworth AE, Hancock JF (2012) Agrobacterium tumefaciens-mediated transformation of rutabaga (Brassica napus var. napobrassica) cultivar “American Purple Top Yellow”. Vitro Cell Dev Biol Plant 48:383–389

    Article  CAS  Google Scholar 

  • Srivastava DK, Kumar P, Sharma S, Gaur A, Gambhir G (2016) Genetic engineering for insect resistance in economically important vegetable crops. In: Ahmad N, Anis M (eds) Plant tissue culture: propagation, conservation and crop. Springer, Berlin, pp 343–378

    Chapter  Google Scholar 

  • Yi D, Cui L, Wang L, Liu Y, Zhuang M, Zhang Y, Zhang J, Lang Z, Zhang Z, Fang Z, Yang L (2013) Pyramiding of Bt cry1Ia8 and cry1Ba3 genes into cabbage (Brassica oleracea L. var. capitata) confers effective control against diamondback moth. Plant Cell Tiss Organ Cult 115:419–428

    Article  CAS  Google Scholar 

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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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Author notes

  1. Geetika Gambhir and Pankaj Kumar have contributed equally to this work.

Authors and Affiliations

  1. ICAR-Indian Agricultural Research Institute, Pusa, New Delhi, 110 012, India

    Geetika Gambhir

  2. CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh, 176 061, India

    Pankaj Kumar

  3. Maharishi Markandeshwar University, Mullana, Ambala, Haryana, 133 207, India

    Gaurav Aggarwal

  4. Dr. Y.S, Parmar University of Horticulture and Forestry, Solan, Himachal Pradesh, 173 230, India

    D. K. Srivastava

  5. ICAR-Directorate of Rapeseed-Mustard Research, Bharatpur, Rajasthan, 321 303, India

    Ajay Kumar Thakur

Authors
  1. Geetika Gambhir
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  2. Pankaj Kumar
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  3. Gaurav Aggarwal
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  4. D. K. Srivastava
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  5. Ajay Kumar Thakur
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Contributions

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.

Corresponding author

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 71, 165–173 (2020). https://doi.org/10.1007/s42977-020-00014-8

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  • DOI: https://doi.org/10.1007/s42977-020-00014-8

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