Chitinolytic assay of indigenous Trichoderma isolates collected from different geographical locations of Chhattisgarh in Central India

Abstract

Chitin is the second most abundant polymer in nature after cellulose and plays a major role in fungal cell walls. As a producer of variety of chitinase enzymes Trichoderma has become an important means of biological control of fungal diseases. A simple and sensitive method based on the use of basal medium with colloidal chitin as sole carbon source supplemented with Bromo cresol purple (pH indicator dye) is proposed to evaluate large populations of Trichoderma for chitinase activity. The soluble substrate with pH indicator dye (Bromo cresol purple, BCP) for the assay of chitinase activity on solid media is sensitive, easy, reproducible semi-quantitative enzyme diffusion plate assay and economic option to determine chitinases. Colloidal chitin derived from Rhizoctonia cell wall and commercial chitin included as a carbon source in broth also allowed selection and comparison of chitinolytic and exochitinase activity in Trichoderma spectrophotometrically. Released N-acetyl-β--D-glucosamine (NAGA) ranged from 37.67 to 174.33 mg/ml and 37.67 to 327.67 mg/ml and p-nitrophenol (pNP) ranged from 0.17 to 35.78 X 10-3 U/ml and 0.62 to 32.6 X 10-3 U/ml) respectively with Rhizoctonia cell wall and commercial chitin derived colloidal chitin supplemented broth.

Trichoderma aureoviride URM 5158 and Trichoderma hamatum URM 6656 are Biocontrol Agents that act against Cassava Root rot through different Mechanisms

“…Agrawal and Kotasthane (2012) indicated that this Trichoderma species presented chitinolytic activity with colloidal chitin supplementation higher than that of T. harzianum, T. viride, T. virens and T. citirinoviride, because T. aureoviride has high affinity to colloidal chitin. Agrawal and Kotasthane (2012) indicated that this Trichoderma species presented chitinolytic activity with colloidal chitin supplementation higher than that of T. harzianum, T. viride, T. virens and T. citirinoviride, because T. aureoviride has high affinity to colloidal chitin.…”

Effect of Trichoderma harzianum on maize rhizosphere microbiome and biocontrol of Fusarium Stalk rot

“…Based on the previous in vitro antagonistic assay, a total of 10 Trichoderma strains Trichoderma asperellum CCTCC-SBW0102 (T3), T. aureoviride CCTCC-SBW0005 (T9), T. harzianum CCTCC-SBW0101 (T11), T. asperellum CCTCC-SBW0109 (T12), T. atroviride CCTCC-SBW0016 (T21), T. asperellum CCTCC-SBW0052 (T43), T. asperellum CCTCC-SBW0091 (T44), T. tawa (T64), T. harzianum CCTCC-RW0024 (T72), and T. harzianum CCTCC-SBW0181 (T88) were selected for the preliminary enzyme assay 41–43 . The percentage of the enzyme activity was calculated using the following formula as Hydrolysis capacity (%) = Diameter of clear zone (cm)/Diameter of Trichoderma colony (cm).…”

Chitinolytic efficacy and secretion of cell wall degrading enzymes from Trichoderma spp. in response to phyto-pathological fung

“…Chitin is a polymer of chitobiose, which consists of two NAGA residues bound to each other by β-1, 4 glycosidic linkage. The active groups present in NAGA molecules are hydroxyl, carbonyl, and imine groups at which anionic dye like BCP is bind and generates color-bound complex (yellow) [2] (pH 4.7) when Trichoderma inoculated in colloidal chitin media having BCP resulted in the formation of chitin into N-acetylglucosamine causing change in pH to more alkalinity and also change in color of indicator dye (BCP) from yellow to purple zone where the chitin got utilized. ) are detected by BCP in the Arrhenius model.…”

“…Genus Trichoderma and efficient biocontrol strain are being industrialized as effective biological fungicides, and their biocontrol mechanism of biological control involves the role of secondary metabolites with prospective applications as new antibiotics [1,2]. They produce many antifungal enzymes including chitin-degrading enzymes which used economically as a basis of these kind of proteins.…”