Planta

, Volume 230, Issue 2, pp 277–291 | Cite as

Defense-related gene expression and enzyme activities in transgenic cotton plants expressing an endochitinase gene from Trichoderma virens in response to interaction with Rhizoctonia solani

  • Vinod Kumar
  • Vilas Parkhi
  • Charles M. Kenerley
  • Keerti S. Rathore
Original Article

Abstract

There are many reports on obtaining disease-resistance trait in plants by overexpressing genes from diverse organisms that encode chitinolytic enzymes. Current study represents an attempt to dissect the mechanism underlying the resistance to Rhizoctonia solani in cotton plants expressing an endochitinase gene from Trichoderma virens. Several assays were developed that provided a powerful demonstration of the disease protection obtained in the transgenic cotton plants. Transgene-dependent endochitinase activity was confirmed in various tissues and in the medium surrounding the roots of transformants. Biochemical and molecular analyses conducted on the transgenic plants showed rapid/greater induction of ROS, expression of several defense-related genes, and activation of some PR enzymes and the terpenoid pathway. Interestingly, even in the absence of a challenge from the pathogen, the basal activities of some of the defense-related genes and enzymes were higher in the endochitinase-expressing cotton plants. This elevated defensive state of the transformants may act synergistically with the potent, transgene-encoded endochitinase activity to confer a strong resistance to R. solani infection.

Keywords

Chitinase Defense Disease resistance Rhizoctonia solani Transgenic cotton Trichoderma virens 

Abbreviations

4-MU-β-(GlucNAc)3

4-Methylumbelliferyl-β-d-N,N′, N″-triacetylchitotrioside

4-MU-β-GlucNAc

4-Methylumbelliferyl-N-acetyl-β-d-glucosaminide

CAD

δ-Cadinene synthase

DAB

3,3′-Diaminobenzidine

GLU

Glucanase

H2DCF-DA

2,7-Dichlorofluorescin diacetate

Hpi

Hours post-inoculation

LOX

Lipoxygenase

MS

Murashige-Skoog

POD

Peroxidase

PR1

Pathogenesis-related protein 1

ROS

Reactive oxygen species

WT

Wild-type

Notes

Acknowledgments

We thank Drs. Robert D. Stipanovic and Lorraine Puckhaber for their help with terpenoid analysis and Drs. Alois Bell and Charles Howell for their valuable suggestions and advice during the course of this investigation. This research was supported by funds from Texas Higher Education Coordinating Board – Advanced Research Program (#000517-0005-2006), Cotton Inc., and Texas AgriLife Research.

Supplementary material

425_2009_937_MOESM1_ESM.pdf (5.4 mb)
Supplementary material 1 (PDF 5479 kb)

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Vinod Kumar
    • 1
  • Vilas Parkhi
    • 1
  • Charles M. Kenerley
    • 3
  • Keerti S. Rathore
    • 1
    • 2
  1. 1.Institute for Plant Genomics and BiotechnologyTexas A&M UniversityCollege StationUSA
  2. 2.Department of Soil and Crop SciencesTexas A&M UniversityCollege StationUSA
  3. 3.Department of Plant Pathology and MicrobiologyTexas A&M UniversityCollege StationUSA

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