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Enhancement of sheath blight tolerance in transgenic rice by combined expression of tobacco osmotin (ap24) and rice chitinase (chi11) genes

  • Genetic Transformation
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Abstract

Agrobacterium-mediated transformation was used to introduce pathogenesis-related protein genes into scutellum-derived callus of rice (Oryza sativa L. subsp. indica ‘White Ponni’) as a means of increasing resistance to sheath blight disease caused by Rhizoctonia solani Kühn. Transformation with a tobacco osmotin (ap24) gene driven by the Cauliflower mosaic virus 35S promoter (P35S) yielded six single-copy transgenic lines. Homozygous T2 plants of all lines accumulated high levels of the ap24 transcript and the osmotin protein and reduced the percent disease index (PDI) of sheath blight disease from 100% down to 49–77%. Transformation with the binary plasmid pNSP2, which harbored the P35S-ap24 gene and a maize ubiquitin promoter (PUbi1)-driven rice chitinase (chi11) gene in the same T-DNA, yielded three single-copy transgenic lines: CO1, CO2, and CO3. Homozygous T2 plants of all three lines accumulated high levels of the chi11 transcript and the chitinase protein. Accumulation of ap24 transcript and osmotin protein was high in homozygous CO1 and CO2, but very low in CO3. At 7 d post infection with R. solani, the transgenic rice lines C18a (with chi11 alone), O8 (with ap24 alone), and CO1, CO2, and CO3 (with chi11 + ap24) reduced the sheath blight PDI from 100% to 64.2, 57.2, 43.3, 50.2, and 58.6%, respectively. At 15 d post infection, the sheath blight disease PDI was significantly lower in the CO1 and CO2 transgenic rice lines, which expressed both chi11 and ap24, than in those which expressed either chi11 or ap24 individually.

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Acknowledgments

We thank Dr. Richard A. Jefferson, Canberra, Australia, for pCAMBIA1301; Dr. Toshihiko Komari, Japan Tobacco, Inc., Shizuoka, Japan, for pSB1; Dr. Stanton B. Gelvin, Purdue University, West Lafayette, Indiana, for Agrobacterium strain LBA4404; Dr. S. Muthukrishnan, Kansas State University, Manhattan, Kansas, for rice chi11 and the chitinase antibody; and Dr. Leo S. Melchers, Mogen International NV, Leiden, The Netherlands, for ap24. We thank Dr. Ray Bressan, Dr. Meena Narasimhan, and Dr. K. G. Raghothama for providing us with the osmotin antibody. We acknowledge Dr. G. Sridevi, Ms. K. Vidhya, and Mr. N. Sabapathi for the construction of pGSD2, pKVD2, and pNSP2, respectively. Program support from the Department of Biotechnology (DBT), Government of India (BT/PR6466/COE/34/16/2012), and the University Grants Commission (UGC), Government of India (No. F. 18-1/2011 [BSR]), BSR Faculty Fellowship to KV are acknowledged. RS is thankful to DBT and UGC for her research fellowships.

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  1. Rajasekaran Sripriya

    Present address: Department of Biotechnology, Mepco Schlenk Engineering College, Sivakasi, Tamil Nadu, 626005, India

  2. Chidambaram Parameswari

    Present address: Department of Plant Breeding and Genetics, Agricultural College and Research Institute, Madurai, Tamil Nadu, 625104, India

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  1. Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, Tamil Nadu, 625021, India

    Rajasekaran Sripriya, Chidambaram Parameswari & Karuppannan Veluthambi

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  1. Rajasekaran Sripriya
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Sripriya, R., Parameswari, C. & Veluthambi, K. Enhancement of sheath blight tolerance in transgenic rice by combined expression of tobacco osmotin (ap24) and rice chitinase (chi11) genes. In Vitro Cell.Dev.Biol.-Plant 53, 12–21 (2017). https://doi.org/10.1007/s11627-017-9807-8

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