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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

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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.

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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

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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.

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Authors and Affiliations

  1. Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, TX, 77843-2123, USA

    Vinod Kumar, Vilas Parkhi & Keerti S. Rathore

  2. Department of Soil and Crop Sciences, Texas A&M University, College Station, TX, 77843-2474, USA

    Keerti S. Rathore

  3. Department of Plant Pathology and Microbiology, Texas A&M University, College Station, TX, 77843-2132, USA

    Charles M. Kenerley

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  1. Vinod Kumar
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  2. Vilas Parkhi
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Correspondence to Keerti S. Rathore.

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Kumar, V., Parkhi, V., Kenerley, C.M. et al. 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 . Planta 230, 277–291 (2009). https://doi.org/10.1007/s00425-009-0937-z

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