Study of Agrobacterium-mediated co-transformation of tea for blister blight disease resistance

Abstract

Blister blight is the most prevalent leaf disease of tea, an economical perennial crop. The disease is caused by a biotrophic fungus, Exobasidium vexans. The objective was set to study whether the combine over-expression of Solanum tuberosum class I chitinase gene and Vigna radiata defensin genes in transgenic tea would give more disease resistance compared to transgene over-expressed singly. Taking two constructs containing S. tuberosum class I chitinase (AF153195) and mung bean defensin (AY437639) gene respectively, three Agrobacterium-mediated transformation events were conducted with two individual constructs and combination of both the constructs. This is a first report about the preparation of transgenic tea by co-transformation for blister resistance. Transformation was successful in all three events. Comparative analysis showed a very interesting result. Minimum overall transformation efficiency was noticed in transformation of tea with pCAMBIA 1301-Chi (2.20%) followed by pBI121-Def (2.39%) and co-transformation with pCAMBIA 1301-Chi and pBI121-Def (3.41%). The fungal bioassay with E. vexans for the transformation event with pCAMBIA 1301-Chi gave the best disease resistance having minimum diameter (7.05 mm) of hypersensitivity zone followed by 15.05 mm in pBI121-Def and 18.14 mm in combination of pCAMBIA 1301-Chi and pBI121-Def.

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Abbreviations

DMSO:

Dimethyl sulfoxide

SEM:

Somatic embryogenesis media

SSE:

Secondary somatic embryo

MM:

Multiplication media

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Acknowledgements

This study was supported by generous funding from Department of Biotechnology, Govt. of India. HRS was Senior Research Fellow supported by Council of Scientific and Industrial Research, Govt. of India. The help from Dr Bornali Gohain during real time PCR work is highly acknowledged. The authors also acknowledge the support of the Director, Tocklai Tea Research Institute, Jorhat, India.

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Correspondence to H. Ranjit Singh.

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Singh, H.R., Hazarika, P., Deka, M. et al. Study of Agrobacterium-mediated co-transformation of tea for blister blight disease resistance. J. Plant Biochem. Biotechnol. 29, 24–35 (2020). https://doi.org/10.1007/s13562-019-00508-0

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Keywords

  • Camellia sinensis
  • Blister blight
  • Exobasidium vexans
  • Potato class I chitinase
  • Mung bean defensin gene
  • Co-transformation