Synergistic activity of chitinases and β-1,3-glucanases enhances fungal resistance in transgenic tomato plants

Jongedijk, Erik; Tigelaar, Henk; Roekel, J. S. C. Van; Bres-Vloemans, Sandra A.; Dekker, Ilma; Elzen, Peter J. M. van den; Cornelissen, Ben J. C.; Melchers, Leo S.

doi:10.1007/978-94-011-0357-2_22

Cited by 80 publications

(64 citation statements)

References 29 publications

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“…There is compelling evidence that ␤-1,3-glucanase and chitinase, acting directly by degrading pathogen cell walls or indirectly by releasing oligosaccharide elicitors of defense reactions, can help defend plants against fungal infection (e.g. Leah et al, 1991;Jach et al, 1995;Jongedijk et al, 1995;Lawrence et al, 1996). Concentrating such a defensive mechanism in the "front-line" tissues of the micropylar endosperm would obviate the need to mount a similar defense throughout the remaining endosperm tissue, particularly because the defense would only be required until the latter had been mobilized to the growing seedling.…”

Section: Discussionmentioning

confidence: 99%

Class I β-1,3-Glucanase and Chitinase Are Expressed in the Micropylar Endosperm of Tomato Seeds Prior to Radicle Emergence

et al. 2001

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β-1,3-Glucanase (EC 3.2.1.39) and chitinase (EC 3.2.1.14) mRNAs, proteins, and enzyme activities were expressed specifically in the micropylar tissues of imbibed tomato (Lycopersicon esculentum Mill.) seeds prior to radicle emergence. RNA hybridization and immunoblotting demonstrated that both enzymes were class I basic isoforms. β-1,3-Glucanase was expressed exclusively in the endosperm cap tissue, whereas chitinase localized to both endosperm cap and radicle tip tissues. β-1,3-Glucanase and chitinase appeared in the micropylar tissues of gibberellin-deficient gib-1tomato seeds only when supplied with gibberellin. Accumulation of β-1,3-glucanase mRNA, protein and enzyme activity was reduced by 100 μM abscisic acid, which delayed or prevented radicle emergence but not endosperm cap weakening. In contrast, expression of chitinase mRNA, protein, and enzyme activity was not affected by abscisic acid. Neither of these enzymes significantly hydrolyzed isolated tomato endosperm cap cell walls. Although both β-1,3-glucanase and chitinase were expressed in tomato endosperm cap tissue prior to radicle emergence, we found no evidence that they were directly involved in cell wall modification or tissue weakening. Possible functions of these hydrolases during tomato seed germination are discussed.

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Section: Discussionmentioning

confidence: 99%

Class I β-1,3-Glucanase and Chitinase Are Expressed in the Micropylar Endosperm of Tomato Seeds Prior to Radicle Emergence

et al. 2001

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“…Transgenic plants with tolerance to fungal pathogens can be developed by transfer and expression of chitinase from homologous or heterologous sources. Although chitinase genes of plant origin were transferred to tomato (Van den Elzen et al, 1993;Jongedijk et al, 1995;Tabaeizadeh et al,1999); the resistance is effective only in a narrow range of pathogens and is quantitatively modest, leading to the need for using highly efficient chitinases to achieve significant levels of plant tolerance. Trichoderma virens is an economically important fungus used commercially as a biofungicide (Mukhopadhyay and Mukherjee, 1996;Howell, 2003) and produces different kinds of chitinases (Kim et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

Expression of a fungal endochitinase gene in transgenic tomato and tobacco results in enhanced tolerance to fungal pathogens

Shah

Mukherjee

Eapen

2010

Physiol Mol Biol Plants

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Development of transgenic Nicotiana tabacum and Lycopersicon esculentum expressing an endochitinase (ech42) gene from biocontrol fungus Trichoderma virens using Agrobacterium-mediated genetic transformation is reported in this paper. Integration of transgene in the genome of transgenic plants was demonstrated using polymerase chain reaction and Southern-blot hybridization, while expression was ascertained by reverse transcription polymerase chain reaction. Histochemical analysis confirmed the expression of GUS enzyme in transformed shoots. Levels of endochitinase enzyme in transgenic plants were found to be up to 10 fold higher compared to control plants. Endochitinase enzyme of 42 kDa was also visualized on SDS-PAGE gel using fluorimetric zymogram in transgenic plants. Endochitinase activity was found to be higher in leaf and stem than the root tissue in transgenic tomato plants. Transgenic lines of both plants showed enhanced resistance to fungal pathogens and a strong negative correlation was found between expression level of endochitinase enzyme and size of disease lesions. Inheritance of transgene, expression and resistance to fungal pathogens of T 1 transgenic tobacco lines was also analysed. The results of the present studies show that ech42 is a promising candidate gene for developing fungal disease resistance in tomato plants.

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“…Early research in other plants had indicated synergistic protective interaction of the coexpression PR proteins. [6][7][8][9][10][11][12] Coexpression chitinase and b-1,3 glucanse by placing both genes in the same T-DNA improved the resistance of tobacco against R. solani. 8 Chitinase and b-1,3 glucanse genes coexpressed in carrot reduced the disease severity caused by Alternaria dauci, A. radicina, C. carotae, and Erysiphe heraclei.…”

Section: Discussionmentioning

confidence: 99%

“…5 To effectively inhibit fungal growth and significantly enhance protection against fungal attack in plants, expression of chitinase genes in combination with other PR-proteins has been reported, which indicated synergistic protective interaction of the coexpressed antifungal proteins. [6][7][8][9][10][11][12] Hybrid tobacco with rice basic chitinase and alfalfa acidic glucanase genes showed at least 75% reduction in the number of lesions produced by Cercospora nicotianae. 7 Transgenic tobacco with barley chitinase and b-1,3 glucanase genes showed enhanced resistance to R. solani.…”

Section: Introductionmentioning

confidence: 99%

“…8 Combined expression tobacco chitinase and b-1,3 glucanase indicated resistance against Fusarium oxysporum, while transgenic tobacco that transformed the two genes, respectively, did not show resistance to F. oxysporum. 9 A modified maize ribosome-inactivating protein and rice chitinase gene in transgenic rice enhanced resistance to sheath blight. 11 Coexpression chi11 and glucanase genes in wheat plants could delay Fusarium head blight development.…”

Section: Introductionmentioning

confidence: 99%

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Purification, characterization of a CkChn134 protein from Cynanchum komarovii seeds and synergistic effect with CkTLP against Verticillium dahliae

et al. 2012

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Cynanchum komarovii Al Iljinski is a desert plant that has been used as analgesic, anthelminthic, and antidiarrheal, but also as herbal medicine to treat cholecystitis in people. In this work, an antifungal protein with sequence homology to chitinase was isolated from C. komarovii seeds and named CkChn134. The three-dimensional structure prediction of CkChn134 indicated that the protein has a loop domain formed a thin cleft, which is able to bind molecules and substrates. The protein and CkTLP synergistically inhibited the fungal growth of Verticillium dahliae, Fusarium oxysporum, Rhizoctonia solani, Botrytis cinerea, and Valsa mali in vitro. The fulllength cDNA was cloned by RT-PCR and RACE-PCR according to the partial protein sequences obtained by nanoESI-MS/MS. The real-time PCR showed that the transcription level of CkChn134 had a significant increase under the stress of ethylene, NaCl, low temperature, drought, and pathogen infection, which indicates that CkChn134 may play an important role in response to abiotic and biotic stresses. The CkChn134 protein was located in the extracellular space/cell wall by CkChn134::GFP fusion protein in transgenic Arabidopsis. Furthermore, overexpression of CkChn134 significantly enhanced the resistance of transgenic Arabidopsis against V. dahliae. Interestingly, the coexpression of CkChn134 and CkTLP showed substantially greater protection against the fungal pathogen V. dahliae than either transgene alone. The results suggest that the CkChn134 is a good candidate protein or gene, and it had a potential synergistic effect with CkTLP for contributing to the development of disease-resistant crops.

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Synergistic activity of chitinases and β-1,3-glucanases enhances fungal resistance in transgenic tomato plants

Cited by 80 publications

References 29 publications

Class I β-1,3-Glucanase and Chitinase Are Expressed in the Micropylar Endosperm of Tomato Seeds Prior to Radicle Emergence

Class I β-1,3-Glucanase and Chitinase Are Expressed in the Micropylar Endosperm of Tomato Seeds Prior to Radicle Emergence

Expression of a fungal endochitinase gene in transgenic tomato and tobacco results in enhanced tolerance to fungal pathogens

Purification, characterization of a CkChn134 protein from Cynanchum komarovii seeds and synergistic effect with CkTLP against Verticillium dahliae

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