Theoretical and Applied Genetics

, Volume 98, Issue 6, pp 1138-1145

Over-expression of the cloned rice thaumatin-like protein (PR-5) gene in transgenic rice plants enhances environmental friendly resistance to Rhizoctonia solani causing sheath blight disease

  • K. DattaAffiliated withPlant Breeding, Genetics and Biochemistry Division, International Rice Research Institute, P.O. Box 3127, 1271 Makati City, Philippines Fax: +63-2-8911292 E-mail: sdatta@irrl.cgnet.com; sdatta@irrl.cgiar.org
  • , R. VelazhahanAffiliated withDepartment of Biochemistry, Kansas State University, Manhattan, KS 66506-3702, USA
  • , N. OlivaAffiliated withPlant Breeding, Genetics and Biochemistry Division, International Rice Research Institute, P.O. Box 3127, 1271 Makati City, Philippines Fax: +63-2-8911292 E-mail: sdatta@irrl.cgnet.com; sdatta@irrl.cgiar.org
  • , I. OnaAffiliated withEntomology and Plant Pathology Division, International Rice Research Institute, P.O. Box 933, Manila 1099, Philippines
  • , T. MewAffiliated withEntomology and Plant Pathology Division, International Rice Research Institute, P.O. Box 933, Manila 1099, Philippines
  • , G. S. KhushAffiliated withPlant Breeding, Genetics and Biochemistry Division, International Rice Research Institute, P.O. Box 3127, 1271 Makati City, Philippines Fax: +63-2-8911292 E-mail: sdatta@irrl.cgnet.com; sdatta@irrl.cgiar.org
  • , S. MuthukrishnanAffiliated withDepartment of Biochemistry, Kansas State University, Manhattan, KS 66506-3702, USA
  • , S. K. DattaAffiliated withPlant Breeding, Genetics and Biochemistry Division, International Rice Research Institute, P.O. Box 3127, 1271 Makati City, Philippines Fax: +63-2-8911292 E-mail: sdatta@irrl.cgnet.com; sdatta@irrl.cgiar.org

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Abstract

 A 1.1-kb DNA fragment containing the coding region of a thaumatin-like protein (TLP-D34), a member of the PR-5 group, was cloned into the rice transformation vector pGL2, under the control of the CaMV 35S promoter. The Indica rice cultivars, ‘Chinsurah Boro II’, ‘IR72’, and ‘IR51500’ were transformed with the tlp gene construct by PEG-mediated direct gene transfer to protoplasts and by biolistic transformation using immature embryos. The presence of the chimeric gene in T0, T1, and T2 transgenic plants was detected by Southern blot analysis. The presence of the expected 23-kDa TLP in transgenic plants was confirmed by Western blot analysis and by staining with Coomassie Brilliant Blue. Bioassays of transgenic plants challenged with the sheath blight pathogen, Rhizoctonia solani, indicated that over-expression of TLP resulted in enhanced resistance compared to control plants.

Key words Thaumatin-like protein PR-5 Rhizoctonia solani Sheath blight disease Rice