For the first time, the specific activities of chitinases, esterases, lipases and a serine protease (VCP1) produced by different isolates of the nematophagous fungus Pochonia chlamydosporia were quantified and compared. The isolates were grown for different time periods in a minimal liquid medium or media supplemented with 1 % chitin, 0.2 % gelatin or 2 % olive oil. Enzyme-specific activities were quantified in filtered culture supernatants using chromogenic p-nitrophenyl substrates (for chitinases, lipases and esterases) and a p-nitroanilide substrate (to measure the activity of the proteinase VCP1). Additionally, information on parasitic growth (nematode egg parasitism) and saprotrophic growth (plant rhizosphere colonisation) was collected. Results showed that the production of extracellular enzymes was influenced by the type of medium (p<0.05) in which P. chlamydosporia was grown. Enzyme activity differed with time (p<0.05), and significant differences were found between isolates (p<0.001) and the amounts of enzymes produced (p<0.001). However, no significant relationships were found between enzyme activities and parasitic or saprotrophic growth using Kendall's coefficient of concordance or Spearman rank correlation coefficient. The results provided new information about enzyme production in P. chlamydosporia and suggested that the mechanisms which regulate the trophic switch in this fungus are complex and dependent on several factors.
The analysis of the functional diversity of soil nematodes requires detailed knowledge on theoretical aspects of the biodiversity–ecosystem functioning relationship in natural and managed terrestrial ecosystems. Basic approaches applied are reviewed, focusing on the impact and value of soil nematode diversity in crop production and on the most consistent external drivers affecting their stability. The role of nematode trophic guilds in two intensively cultivated crops are examined in more detail, as representative of agriculture from tropical/subtropical (banana) and temperate (apple) climates. The multiple facets of nematode network analysis, for management of multitrophic interactions and restoration purposes, represent complex tasks that require the integration of different interdisciplinary expertise. Understanding the evolutionary basis of nematode diversity at the field level, and its response to current changes, will help to explain the observed community shifts. Integrating approaches based on evolutionary biology, population genetics and ecology can quantify the contribution of nematode fauna to fundamental soil functions. These include carbon transformation, nutrient cycling, pest control and disease transmission. In conclusion, different facets of nematode diversity such as trophic groups, life history traits, variability in body size and/or taxa identities in combination with DNA-based techniques are needed in order to disclose nematode–soil–ecosystem functioning relationships. Further experimental studies are required to define locally adapted and sustainable management practices, through ecosystem-based approaches and nature-based solutions.
The fungus Pochonia chlamydosporia is a potential biological control agent for plant parasitic nematodes, but to date, there has been little investigation of interactions (competitive, antagonistic or synergistic) between different isolates that occur together on roots and nematode galls. Real-time quantitative PCR (qPCR) has greatly improved the study of many fungi in situ on plant and nematode hosts, but distinguishing closely related isolates remains difficult. In this study, primers to discriminate P. chlamydosporia var. chlamydosporia and P. chlamydosporia var. catenulata were used to measure the relative abundance of isolates of the two varieties when inoculated singly or together on tomato plants. Also, sequence-characterised amplified polymorphic regions were identified to distinguish two different isolates of P. chlamydosporia var. chlamydosporia. Individual 1-cm root segments and nematode galls were excised, DNA extracted and subjected to real-time qPCR with the discriminatory primers. The qPCR method proved sensitive and reproducible and demonstrated that roots and nematode galls were not uniformly colonised by the fungi. Results indicated that the P. chalmydosporia var. catenulata isolate was more abundant on roots and eggs than P. chlamydosporia var. chlamydosporia, but all the isolates infected a similar proportion of nematode eggs. There was an indication that the abundance of each fungal isolate was reduced in coinoculation experiments compared with single inoculations, but the number of root segments and galls colonised was not statistically significantly different.
Species of the genus Nacobbus have the potential to reduce yields of major food crops such as potato, sugar beet and tomato in many parts of the world, thus warranting a quarantine effort to avoid their introduction. Molecular studies offer a new method for routine quarantine diagnostics for this nematode that will be faster and more sensitive than previous methods. A primer set was designed from Nacobbus ITS sequences and their specificity confirmed. DNA was extracted from nematodes, soil and potato tubers for use in PCR. Optimised PCR conditions were established and the PCR products were separated on 2% agarose gels, showing that specific ITS primers for the detection of Nacobbus generated a single PCR product, although band size varied slightly between species and soil isolates. The product was generated from DNA extracted from all the Nacobbus samples but not from other nematodes tested (Pratylenchus, Radopholus, Meloidogyne, Globodera, Heterodera). No bands were generated from the uninfested control soil and control tuber DNA samples, thus demonstrating the specificity of the primers. For the first time, Nacobbus was detected in soil and tuber samples using molecular approaches. These results have important applications not only in analysing advisory samples but also in the screening of material for quarantine purposes.
For the first time, the effects of varying osmotic and matric potential on fungal radial growth and accumulation of polyols were studied in three isolates of Pochonia chlamydosporia. Fungal radial growth was measured on potato dextrose agar modified osmotically using potassium chloride or glycerol. PEG 8000 was used to modify matric potential. When plotted, the radii of the colonies were found to grow linearly with time, and regression was applied to estimate the radial growth rate (mm day (1 ). Samples of fresh mycelia from 25-day-old cultures were collected and the quantity (mg g (1 fresh biomass) of four polyols (glycerol, erythritol, arabitol and mannitol) and one sugar (glucose) was determined using HPLC. Results revealed that fungal radial growth rates decreased with increased osmotic or matric stress. Statistically significant differences in radial growth were found between isolates in response to matric stress (PB0.006) but not in response to osmotic stress (P00.759). Similarly, differences in the total amounts of polyols accumulated by the fungus were found between isolates in response to matric stress (PB0.001), but not in response to osmotic stress (P00.952). Under water stress, the fungus accumulated a combination of different polyols important in osmoregulation, which depended on the solute used to generate the stress. Arabitol and glycerol were the main polyols accumulated in osmotically modified media, whereas erythritol was the main polyol that was accumulated in media amended with PEG. The results found that Pochonia chlamydosporia may use different osmoregulation mechanisms to overcome osmotic and matric stresses.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.