Heterologous expression of new antifungal chitinase from wheat

Singh, Arpita; Kirubakaran, S. Isaac; Sakthivel, Natarajan

doi:10.1016/j.pep.2007.06.013

Cited by 59 publications

(71 citation statements)

References 51 publications

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“…Of these PR proteins, the PR-3 chitinases were well represented in the intermediate fractions, as both precursors and mature proteins. PR-3 chitinases are fungal growth inhibitor proteins, which hydrolyze the chitin of the fungal cell walls and so disrupt hyphal entry to the live inner fraction and endosperm (Selitrennikoff, 2001;Singh et al, 2007). However, the PR-2 class II chitinase (b-1,3-glucanase) identified here and in the inner layer has a different function (Mauch and Staehelin, 1989).…”

Section: Discussionmentioning

confidence: 96%

“…These elicitor molecules induce the expression of both PR-2 and PR-3 proteins, which are concentrated in the vacuole. The vacuole bursts when fungal hyphae penetrate the cell, releasing a high concentration of these chitinases (Mauch and Staehelin, 1989;Singh et al, 2007). The PR proteins wheatwin1 (PR-4) and thaumatin-like protein (PR-5) are also both inducible as part of a chemically elicited defense response (Desmond et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

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Strategic Distribution of Protective Proteins within Bran Layers of Wheat Protects the Nutrient-Rich Endosperm

et al. 2010

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Bran from bread wheat (Triticum aestivum ‘Babbler’) grain is composed of many outer layers of dead maternal tissues that overlie living aleurone cells. The dead cell layers function as a barrier resistant to degradation, whereas the aleurone layer is involved in mobilizing organic substrates in the endosperm during germination. We microdissected three defined bran fractions, outer layers (epidermis and hypodermis), intermediate fraction (cross cells, tube cells, testa, and nucellar tissue), and inner layer (aleurone cells), and used proteomics to identify their individual protein complements. All proteins of the outer layers were enzymes, whose function is to provide direct protection against pathogens or improve tissue strength. The more complex proteome of the intermediate layers suggests a greater diversity of function, including the inhibition of enzymes secreted by pathogens. The inner layer contains proteins involved in metabolism, as would be expected from live aleurone cells, but this layer also includes defense enzymes and inhibitors as well as 7S globulin (specific to this layer). Using immunofluorescence microscopy, oxalate oxidase was localized predominantly to the outer layers, xylanase inhibitor protein I to the xylan-rich nucellar layer of the intermediate fraction and pathogenesis-related protein 4 mainly to the aleurone. Activities of the water-extractable enzymes oxalate oxidase, peroxidase, and polyphenol oxidase were highest in the outer layers, whereas chitinase activity was found only in assays of whole grains. We conclude that the differential protein complements of each bran layer in wheat provide distinct lines of defense in protecting the embryo and nutrient-rich endosperm.

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Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Strategic Distribution of Protective Proteins within Bran Layers of Wheat Protects the Nutrient-Rich Endosperm

et al. 2010

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“…However, transgenic tobacco and wheat plants expressing a barley class II chitinase showed enhanced resistance to fungal infection (Jach et al 1995;Oldach et al 2001). The involvement of the cysteine-rich domain in antifungal efficacy has yet to be definitively characterized (Singh et al 2007). Class II chitinase was reported from bark tissue of stems V. corymbosum, in the Chitinase activity of fresh nectar with chitin azure.…”

Section: Discussionmentioning

confidence: 99%

Identification and cloning of class II and III chitinases from alkaline floral nectar of Rhododendron irroratum, Ericaceae

Zha

Milne

Zhou

et al. 2016

Planta

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Class II and III chitinases belonging to different glycoside hydrolase families were major nectarins in Rhododendron irroratum floral nectar which showed significant chitinolytic activity. Previous studies have demonstrated antimicrobial activity in plant floral nectar, but the molecular basis for the mechanism is still poorly understood. Two chitinases, class II (Rhchi2) and III (Rhchi3), were characterized from alkaline Rhododendron irroratum nectar by both SDS-PAGE and mass spectrometry. Rhchi2 (27 kDa) and Rhchi3 (29 kDa) are glycoside hydrolases (family 19 and 18) with theoretical pI of 8.19 and 7.04. The expression patterns of Rhchi2 and Rhchi3 were analyzed by semi-quantitative RT-PCR. Rhchi2 is expressed in flowers (corolla nectar pouches) and leaves while Rhchi3 is expressed in flowers. Chitinase in concentrated protein and fresh nectar samples was visualised by SDS-PAGE and chitinolytic activity in fresh nectar was determined spectrophotometrically via chitin-azure. Full length gene sequences were cloned with Tail-PCR and RACE. The amino acid sequence deduced from the coding region for these proteins showed high identity with known chitinases and predicted to be located in extracellular space. Fresh R. irroratum floral nectar showed significant chitinolytic activity. Our results demonstrate that class III chitinase (GH 18 family) also exists in floral nectar. The functional relationship between class II and III chitinases and the role of these pathogenesis-related proteins in antimicrobial activity in nectar is suggested.

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“…(grain discoloration of rice), and Fusarium sp. (scab of rye) (Singh et al 2007). Therefore, due to the innate antifungal potential, wheat chitinase can be used to enhance fungal resistance in crop plants.…”

Section: Introductionmentioning

confidence: 99%

Transgenic tomato plants expressing a wheat endochitinase gene demonstrate enhanced resistance to Fusarium oxysporum f. sp. lycopersici

Girhepuje

Shinde

2010

Plant Cell Tiss Organ Cult

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Various chitinases have been shown to inhibit the growth of fungal pathogens in in vitro as well as in planta conditions. chi194, a wheat chitinases gene encoding a 33-kDa chitinase protein, was overexpressed in tomato plants (cv. Pusa Ruby) under the control of maize ubiquitin 1 promoter. The integration of transgene in tomato plants was confirmed with polymerase chain reaction (PCR) and Southern blot analysis. The inheritance of the transgene in T 1 and T 2 generations were shown by molecular analysis and the hygromycin sensitivity test. The broad range of chitinase activity was observed among the transgenic lines in T 0 and a similar range was retained in the T 1 and T 2 generations. Most importantly, the transgenic tomato lines with high chitinase activity were found to be highly resistant to the fungal pathogen Fusarium oxysporum f. sp. lycopersici. Thus, the results demonstrated that the expression of the wheat endochitinase chi194 in tomato plants confers resistance against Fusarium wilt disease caused by the fungal pathogen Fusarium oxysporum f. sp. lycopersici.

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Heterologous expression of new antifungal chitinase from wheat

Cited by 59 publications

References 51 publications

Strategic Distribution of Protective Proteins within Bran Layers of Wheat Protects the Nutrient-Rich Endosperm

Strategic Distribution of Protective Proteins within Bran Layers of Wheat Protects the Nutrient-Rich Endosperm

Identification and cloning of class II and III chitinases from alkaline floral nectar of Rhododendron irroratum, Ericaceae

Transgenic tomato plants expressing a wheat endochitinase gene demonstrate enhanced resistance to Fusarium oxysporum f. sp. lycopersici

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