scholarly journals In vitro screening of new strains of predacious nematophagous fungi for biocontrol suitability when produced in liquid culture

2021 ◽  
Vol 8 (2) ◽  
pp. 3-15
Author(s):  
H. Tkalenko ◽  
Ya. Gadzalo ◽  
O. Borzykh ◽  
S. Horal
Keyword(s):  
Mass Production ◽  
Nematophagous Fungi ◽  
Submerged Cultivation ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Free Living ◽  
Predacious Fungi ◽  
First Time ◽  
Different Soils

Aim. To isolate, identify, and evaluate in vitro the predacious ability of nematophagous fungi strains, to determine their eligibility for mass production under submerged cultivation; to select the most efficient isolates of predacious nematopha- gous fungi as promising producers of the biopreparation to control plant-parasitic nematodes. Methods. Microbiological, microscopical, cultural-morphological, statistical. Results. The screening to determine the presence of predacious nema- tophagous fungi in different soils from different regions has been conducted for the first time in Ukraine. Out of 75 soil samples, isolated in Kyiv, Chernihiv, Dnipropetrovsk, Volyn and Odesa regions, 88 isolates of predacious nematophagous fungi were obtained which belong to 11 genera by their cultural-morphological characteristics: Arthrobotrys oligospo- ra – 39 isolates, A. musiformis – 25, A. conoides – 11, Drechslerella dactyloides – 3, A. artrobotryoides –2, A. superba – 2, A. megalospora – 2, A. sphaeroides –1, A. flagrans – 1, A. amerospora – 1, A. thaumasia – 1. High nematophagous activity in vitro (90–100 % trapping) regarding free-living nematodes (Rhabditis spp). was demonstrated by 4 isolates of A. oligospora, 18 isolates of A. musiformis, 5 isolates of A. conoides, 3 isolates of D. dactyloides, and one isolate of A. megalospora. Among the 47 most active isolates, 4 isolates of predacious fungi formed chlamydospores under submerged cultivation on two types of liquid media – wort medium and corn-molasses medium: A. musiformis-711, A. musiformis-911, A. conoides-90, D. dactyloides-19. It was found that isolate A. musiformis-911 and A. conoides-90 could form conidia in the submerged culture. Further experiments in determining the character of development for two latter isolates in different types of media under submerged cultivation established that the largest number of chlamydo- spores and conidia were formed by the isolates of A. conoides 90 and A. musiformis 911 on potato dextrose medium, 7.6·105 and 6.5·105 spores/ml respectively. In the wort medium, the productivity of spore formation for the isolate of A. conoides 90 was 35 times lower, amounting to 2.2·104 spores/ml, and in peptone-glucose medium – 90 times lower (8.5·103 spores/ml). The isolate of A. musiformis 911 in the wort medium produced 325 times fewer spores (2.0× ×103 spores/ml) than in potato-dextrose medium, while no chlamydospores and conidia were formed in the peptone- glucose medium. More detailed study of the ratio between isolates-producers and different sources of nutrition as the basis for the optimization of liquid media is required. Conclusions. According to the results of determining nematoph- agous activity regarding free-living nematodes, isolated from different soils in Ukraine and evaluating the character of development under submerged cultivation, two isolates of predacious fungi were selected – Arthrobotrys conoides-90 and A. musiformis-911, which are promising producers of a biological preparation eventually to be used in the biocon- trol of plant-parasitic nematodes. Conidia formation of the two isolates in liquid culture under mass production condi- tions was observed for the first time and could also contribute to their suitability for mass production and biocontrol.

Biological attributes of Salibro™, a novel sulfonamide nematicide. Part 2: Impact on the fitness of various non-target nematodes

Nematology ◽  
2020 ◽  
pp. 1-17 ◽  
Author(s):  
Tim C. Thoden ◽  
Mariam A. Alkader ◽  
John A. Wiles
Keyword(s):  
Negative Impact ◽  
Soil Health ◽  
Agar Plate ◽  
Parasitic Nematodes ◽  
Published Data ◽  
Plant Parasitic Nematodes ◽  
Free Living ◽  
Plant Parasitic ◽  
Novel Products

Summary Currently a renaissance in chemical nematicides is taking place with novel products like Nimitz® (a.s. fluensulfone), Velum Prime® (a.s. fluopyram) and Salibro™ (a.s. fluazaindolizine – Reklemel™ active) entering the marketplace. Although a considerable amount of published data is already available on their laboratory and field impact on plant-parasitic nematodes, little is understood of their compatibility with the beneficial or free-living nematodes that are part of the soil health network. In a range of laboratory studies, the effects of these nematicides on the vitality and reproduction of several species was tested, including both cosmopolitan free-living nematodes (Acrobeloides, Cruznema, Panagrobelus) as well as commercially applied entomopathogenic nematodes (Steinernema, Heterorhabditis). Within aqueous exposure and agar plate in vitro assays, species sensitivity to those nematicides differed significantly but their fitness (vitality and reproduction; infectivity to insect hosts) was generally not adversely impacted by concentrations of 5-50 ppm (a.s.) of Salibro. Even at 250 ppm (a.s.) of Salibro only some species of the bacterial-feeding species showed some negative impact. By contrast, both Nimitz at 50 ppm (a.s.) and Velum at 5 ppm (a.s.) consistently demonstrated stronger adverse impacts. In second level soil drenching assays, either no, or occasionally slight, adverse effects on the natural community of free-living nematodes were observed if soils were drenched with different volumes of Salibro at 5-50 ppm (a.s.), while relatively stronger reductions were measured within the plant-parasitic species (especially root-knot nematodes). Both Nimitz and Vydate (a.s. oxamyl) showed some degree of compatibility at 5 and 25 ppm (a.s.), respectively, which was generally higher than for Velum Prime at 5 ppm (a.s.). Overall, these data indicate that, when used at common field rates, Salibro will be one of the best options as part of integrated nematode management programmes where the use of chemical nematicides is required.

scholarly journals The Ditylenchus destructor genome provides new insights into the evolution of plant parasitic nematodes

2016 ◽  
Vol 283 (1835) ◽  
pp. 20160942 ◽  
Author(s):  
Jinshui Zheng ◽  
Donghai Peng ◽  
Ling Chen ◽  
Hualin Liu ◽  
Feng Chen ◽  
...  
Keyword(s):  
Evolutionary History ◽  
Evolutionary Process ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Free Living ◽  
C Elegans ◽  
Free Living Nematode ◽  
Plant Nematode ◽  
Ditylenchus Destructor ◽  
Plant Parasitic

Plant-parasitic nematodes were found in 4 of the 12 clades of phylum Nematoda. These nematodes in different clades may have originated independently from their free-living fungivorous ancestors. However, the exact evolutionary process of these parasites is unclear. Here, we sequenced the genome sequence of a migratory plant nematode, Ditylenchus destructor . We performed comparative genomics among the free-living nematode, Caenorhabditis elegans and all the plant nematodes with genome sequences available. We found that, compared with C. elegans , the core developmental control processes underwent heavy reduction, though most signal transduction pathways were conserved. We also found D. destructor contained more homologies of the key genes in the above processes than the other plant nematodes. We suggest that Ditylenchus spp. may be an intermediate evolutionary history stage from free-living nematodes that feed on fungi to obligate plant-parasitic nematodes. Based on the facts that D. destructor can feed on fungi and has a relatively short life cycle, and that it has similar features to both C. elegans and sedentary plant-parasitic nematodes from clade 12, we propose it as a new model to study the biology, biocontrol of plant nematodes and the interaction between nematodes and plants.

scholarly journals Defining the early stages of intestinal colonisation by whipworms

2020 ◽  
Author(s):  
María A. Duque-Correa ◽  
David Goulding ◽  
Claire Cormie ◽  
Catherine Sharpe ◽  
Judit Gali Moya ◽  
...  
Keyword(s):  
Intestinal Epithelium ◽  
Early Stage ◽  
Proximal Colon ◽  
Host Cells ◽  
Parasitic Nematodes ◽  
Metazoan Parasites ◽  
Multiple Cells ◽  
First Time

ABSTRACTHundreds of millions of people are infected with whipworms (Trichuris trichiura), large metazoan parasites that live in the caecum and proximal colon. Whipworms inhabit distinct multi-intracellular epithelial burrows that have been described as syncytial tunnels. However, the interactions between first-stage (L1) larvae and the host epithelia that determine parasite invasion and establishment in the syncytium remain unclear. In vivo experiments investigating these events have been severely hampered by the limited in situ accessibility to intracellular infective larvae at the bottom of the crypts of Lieberkühn, and the lack of genetic tools such as fluorescent organisms that are readily available for other pathogens but not parasitic nematodes. Moreover, cell lines, which do not mimic the complexity of the intestinal epithelium, have been unsuccessful in supporting infection by whipworm larvae. Here, we show that caecaloids grown in an open crypt-like conformation recapitulate the caecal epithelium. Using this system, we establish in vitro infections with T. muris L1 larvae for the first-time, and provide clear evidence that syncytial tunnels are formed at this early stage. We show that larval whipworms are completely intracellular but woven through multiple cells. Using the caecaloids, we are able to visualise the pathways taken by the larvae as they burrow through the epithelial cells. We also demonstrate that larvae degrade the mucus layers overlaying the epithelium, enabling them to access the cells below. We show that early syncytial tunnels are composed of enterocytes and goblet cells that are alive and actively interacting with the larvae during the first 24 h of the infection. Progression of infection results in damage to host cells and by 72 h post-infection, we show that desmosomes of cells from infected epithelium widen and some host cells appear to become liquified. Collectively, our work unravels processes mediating the intestinal epithelium invasion by whipworms and reveals new specific interactions between the host and the parasite that allow the whipworm to establish on its multi-intracellular niche. Our study demonstrates that caecaloids can be used as a relevant in vitro model to investigate the infection biology of T. muris during the early colonisation of its host.

Combinatorial in vitro RNAi of two neuropeptide genes and a pharyngeal gland gene on Meloidogyne incognita

Nematology ◽  
2015 ◽  
Vol 17 (2) ◽  
pp. 155-167 ◽  
Author(s):  
Prakash Banakar ◽  
Amita Sharma ◽  
Catherine J. Lilley ◽  
Nagavara Prasad Gantasala ◽  
Mukesh Kumar ◽  
...  
Keyword(s):  
Meloidogyne Incognita ◽  
Parasitic Nematodes ◽  
Specific Gene ◽  
Plant Parasitic Nematodes ◽  
Rnai Silencing ◽  
Transcript Quantification ◽  
The Central Nervous System ◽  
Pharyngeal Gland

Root-knot nematodes are the most economically important group of plant-parasitic nematodes. In the present study, functional validation using in vitro RNAi was carried out on Meloidogyne incognita with two FMRFamide-like peptide genes, flp-14 and flp-18, and a subventral pharyngeal gland specific gene, 16D10. It was found that RNAi silencing of each gene reduced the attraction of M. incognita at different time intervals both in combination and individually. Silencing of the genes reduced nematode infection by 23-30% and development as indicated by a reduction in the number of females by 26-62%. Reproduction was decreased by 27-73% and fecundity was decreased by 19-51%. In situ hybridisation revealed the expression of flp-18 in cells associated with the ventral and retro vesicular ganglia of the central nervous system. qRT-PCR supported the correlation between phenotypic effects of silencing with that of transcript quantification.

Nematophagous Fungi in Antioxidant-Mediated Defense Against Plant Parasitic Nematodes

2021 ◽  
pp. 607-622
Author(s):  
S. Saranya ◽  
Basavaraj Teli ◽  
Jyotika Purohit ◽  
R. K. Singh ◽  
Anirudha Chattopadhyay
Keyword(s):  
Nematophagous Fungi ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Plant Parasitic

scholarly journals Differential Response of Meloidogyne, Pratylenchus, Globodera, and Xiphinema Species to the Nematicide Fluazaindolizine

2020 ◽  
Vol 110 (12) ◽  
pp. 2003-2009
Author(s):  
Catherine L. Wram ◽  
Inga Zasada
Keyword(s):  
Untreated Control ◽  
Exposure Dose ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Response Curves ◽  
Greenhouse Study ◽  
Meloidogyne Spp ◽  
The Impact ◽  
Plant Parasitic

This research focused on the effects of fluazaindolizine on a diversity of plant-parasitic nematodes. In microwell assays, 24-h dose-response curves were generated for several species and populations of Meloidogyne, Pratylenchus neglectus, P. penetrans, Globodera ellingtonae, and Xiphinema americanum. In a greenhouse study, the impact of fluazaindolizine on fecundity of M. incognita, M. hapla, and M. chitwoodi was tested by exposing nematodes for 24 h in solution and inoculating on tomato. The average 24-h ED50s (dose that resulted in the immobility of 50% of exposed nematodes) for M. hapla, M. chitwoodi, and M. incognita were 325.7, 223.4, and 100.7 ppm, respectively. M. hapla had the most variation among populations, with 24-h ED50s ranging from 72 to 788 ppm. G. ellingtonae had the lowest 24-h ED50 at 30 ppm. Pratylenchus spp. were unaffected by fluazaindolizine. X. americanum was the only species where effects of fluazaindolizine were reversible, but had a 24-h ED50 that fell in the range of the Meloidogyne spp. In the greenhouse study, M. chitwoodi was the least sensitive with reproduction reaching 62% of the untreated control after a pre-exposure to 47 ppm, whereas M. incognita and M. hapla at the same exposure dose had reproduction rates of 27 and 36% of the untreated control, respectively. Despite varying in in vitro responses to fluazaindolizine, reproduction of all Meloidogyne spp. was suppressed after only 24 h of exposure. This study expanded our understanding of how G. ellingtonae, P. thornei, P. penetrans, and X. americanum respond to fluazaindolizine.

scholarly journals Activity of Saponins from Medicago Species against Phytoparasitic Nematodes

Plants ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 443 ◽  
Author(s):  
Trifone D’Addabbo ◽  
Maria Pia Argentieri ◽  
Jerzy Żuchowski ◽  
Elisa Biazzi ◽  
Aldo Tava ◽  
...  
Keyword(s):  
Meloidogyne Incognita ◽  
In Vitro Assays ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Phytoparasitic Nematodes ◽  
Phytochemical Profile ◽  
Medicago Species ◽  
Phytoparasitic Nematode ◽  
Plant Parasitic

Content of bioactive saponins of Medicago species suggests that they may also exert, as previously demonstrated on M. sativa, nematicidal properties exploitable for the formulation of new products for sustainable phytoparasitic nematode management. This study was addressed to highlight the bioactivity of saponins from five different Medicago species still poorly known for their biological efficacy, i.e., M. heyniana, M. hybrida, M. lupulina, M. murex and M. truncatula, against the plant parasitic nematodes Meloidogyne incognita, Xiphinema index and Globodera rostochiensis. The bioactivity of the extracts from the five Medicago species was assessed by in vitro assays on the juveniles (J2) and eggs of M. incognita and G. rostochiensis and the adult females of X. index. The suppressiveness to M. incognita of soil treatments with the Medicago plant biomasses was also investigated in a tomato experiment. The nematicidal activity of the five Medicago species was reported and discussed in relation to their phytochemical profile.

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