Inoculation of Pochonia chlamydosporia triggers a defense response in tomato roots, affecting parasitism by Meloidogyne javanica.

Pochonia chlamydosporia is a soil-dwelling fungus and biological control agent of nematodes, active ingredient in commercial bionematicides. The fungus is also endophytically associated with the roots of several plant species, promoting their growth and inducing systemic resistance. In this study, different pathways and tomato defense metabolites were studied to identify mechanisms induced by P. chlamydosporia that contribute to the control of Meloidogyne javanica, at early and late developmental stages. Some defense responses activated by the fungus appeared related to the nematode life cycle. Among the evaluated biochemical analysis, root colonization of P. chlamydosporia showed an increase in the concentration of phenolic compounds, such as chlorogenic acid. In addition, the expression of some host plant genes was also modified. The interaction of the fungus with roots parasitized by M. javanica resulted in the highest expression of Phenylalanine Ammonia-Lyase (PAL), Chalcone synthase (LECHS 2), and Protease Inhibitor (PI1) genes at 24 days post-inoculation. At the second sampling time (44 days), there was an increase in the expression of the Respiratory Burst Oxidase Homolog (RBOH) gene. Fungus reduced the expression of the ACC-oxidase and Pathogenesis-Related Proteins 1 (PR-1) genes in roots. Moreover, P. chlamydosporia inoculation changed metabolites and phytohormone profiles of the gall formed by M. javanica. Plant defense response appeared to involve the jasmonic acid and phytosphingosine cascades. With this analysis, it was possible to propose new molecular mechanisms induced by the fungus that contribute to the control of M. javanica.

In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia.

The objective of this study was to purify, characterize, and phylogenetically and structurally analyze the dextranase produced by the fungus Pochonia chlamydosporia. Dextranase produced by the fungus P. chlamydosporia was purified to homogeneity in two steps, with a yield of 152%, purification factor of 6.84 and specific activity of 358.63 U/mg. Its molecular weight was estimated by SDS-PAGE at 64 kDa. The enzyme presented higher activity at 50 °C and pH 5.0, using 100 mM citrate-phosphate buffer, was inhibited by Ag1+, Hg2+, Cu2+, Mg2+, and presented KM of 23.60 µM. Mature dextranase is composed of 585 amino acids residues, with a predicted molecular weight of 64.38 kDa and pI 5.96. This dextranase showed a strong phylogenetic similarity when compared to Trichoderma harzianum dextranase. Its structure consists of two domains: the first composed by 15 ß strands, and the second composed by a right-handed parallel ß-helix.

Nematocidal activity of extracellular enzymes produced by the nematophagous fungus Duddingtonia flagrans on cyathostomin infective larvae.

Duddingtonia flagrans produces chitinases, however, optimization of the production of these enzymes still needs to be explored, and its nematocidal activity should still be the subject of studies. The objective of the present study was to optimize chitinase production, and evaluate the nematocidal activity of extracellular enzymes produced by the nematophagous fungus D. flagrans on cyathostomin infective larvae. An isolate from D. flagrans (AC001) was used in this study. For the production of enzymes (protease and chitinase), two different culture media were inoculated with AC001 conidia. Both enzymes were purified. The statistical Plackett-Burman factorial design was used to investigate some variables and their effect on the production of chitinases by D. flagrans. After that, the design central composite (CCD) was used in order to determine the optimum levels and investigate the interactions of these variables previously observed. Only two variables (moisture and incubation time), in the evaluated levels, had a significant effect (p<0.05) on chitinase production. The conditions of maximum chitinase activity were calculated, with the following values: incubation time 2 days, and moisture 511%. The protease and chitinase derived from D. flagrans, individually or together (after 24h), led to a significant reduction (p<0.01) in the number of intact cyathostomin L3, when compared to the control, with following reduction percentage values: 19.4% (protease), 15.5% (chitinase), and 20.5% (protease+chitinase). Significant differences were observed (p<0.05) between the group treated with proteases in relation to the group treated with proteases+chitinases. In this study, the assay with the cyathostomins showed that chitinase had a nematocidal effect, suggesting that this enzyme acts on the "fungus versus nematodes" infection process. It is known that nematode eggs are rich in chitin, and in this case, we could think of a greater employability for this chitinase.