Chitinase and β-1,3-glucanase enzyme production by the mycoparasiteClonostachys roseaf.catenulataagainst fungal plant pathogens

Abstract

Clonostachys rosea f. catenulata (syn. Gliocladium catenulatum) is an effective fungal biological agent against Fusarium root and stem rot and Pythium damping-off diseases on cucumber plants. Both chitinase and beta-1,3-glucanase enzymes were produced when C. rosea was grown on a synthetic medium containing chitin or laminarin as a sole carbon source, respectively. Chitinase production was also induced by Fusarium cell walls, while beta-1,3-glucanase activity was induced by both Fusarium and Pythium cell walls, as well as by growth on homogenized cucumber roots and on low-carbon media. Mycelial growth of Fusarium and Pythium, when exposed to C. rosea culture filtrates that contain glucanase activity, was significantly reduced compared with the controls, and cell walls of both pathogens were degraded. On excised cucumber roots, hyphae of C. rosea formed appressorium-like structures and coiled around hyphae of Pythium. In culture, C. rosea caused localized degradation of Fusarium hyphae. Cucumber root tissues colonized by C. rosea showed higher levels of beta-1,3-glucanase activity at 7 days post-application compared with untreated controls. To determine if this activity was derived from C. rosea, glucanase isoforms were separated on activity gels. Fungal culture filtrates and root extracts contained the same predominant 20 kDa isoform. Reverse-transcription polymerase chain reaction (RT-PCR) using primers designed to amplify a beta-1,3-glucanase gene in C. rosea confirmed glucanase expression on roots. These results show that C. rosea produces beta-1,3-glucanase in situ, which can degrade hyphae of Fusarium and Pythium and contribute to biological control efficacy.

Transformation of the endochitinase gene Chi67-1 in Clonostachys rosea 67-1 increases its biocontrol activity against Sclerotinia sclerotiorum

“…catenulate (syn. Gliocladium catenulatum ), is currently mass produced and applied to vegetables, herbs and ornamentals in greenhouses and fields (Chatterton and Punja 2009; Gwynn 2014; Punja and Yip 2003; Rahman and Punja 2007). Thus far, problems such as the unstable efficiency of fungal biopesticides in field applications have limited their development, and the pathogenicity of most wild type isolates needs to be improved to achieve a higher control efficiency.…”

Colonization of geranium foliage by Clonostachys rosea f. catenulata, a biological control agent of botrytis grey mould

“…Efficacy of C. rosea f. catenulata in suppressing rootinfecting pathogens has been attributed to both its root colonization ability and production of cell wall degrading enzymes in situ, resulting in reduced sporulation and establishment of fungal propagules (Chatterton et al 2008;Chatterton and Punja 2009). The ability of C. rosea f. catenulata to produce cell wall degrading enzymes on foliar tissues and how this impacts foliar pathogens remains to be determined.…”

In vitro and In vivo Antifungal Activity of Culture Filtrates and Organic Extracts of Penicillium sp. and Gliocladium spp. against Botrytis cinerea

“…Brunner et al [34] also found active compounds in the culture filtrates of T. atroviride, such as glucose oxidase, able to inhibit B. cinerea spore germination. In the same sense, Chatterton [35] showed that chitinase and glucanase contained in the culture filtrate of G. catenulatum can inhibit by 50% F. oxysporum and Pythium spp. mycelial growth.…”

“…secondary metabolites were ethyl acetate and hexane. Chatterton [35] found that the enzymatic extracts of G. catenulatum were found to be effective agents depends on the concentrations used. In fact, they demonstrated that increasing the concentration of the filtrate obtained from Rhodotorula glutinis led to a limited incidence of the grey mold in strawberries.…”