Culturing techniques.

Author(s):  
Rosa H. Manzanilla-López ◽  
Ralf-Udo Ehlers

Abstract A nematological culture is a method of multiplying nematodes by enabling them to reproduce and increase in a culture medium or a specific host under controlled conditions. This chapter presents culturing techniques, focused mainly on plant-parasitic nematodes and entomopathogenic nematodes, with some mention of soil free-living nematodes.

Nematology ◽  
1999 ◽  
Vol 1 (7) ◽  
pp. 735-743 ◽  
Author(s):  
Parwinder S. Grewal ◽  
Edwin E. Lewis ◽  
Sudha Venkatachari

Abstract A possible mechanism of suppression of a plant-parasitic nematode Meloidogyne incognita by entomopathogenic nematodes is described. Heat-killed entomopathogenic nematodes Steinernema feltiae and S. riobrave temporarily suppressed penetration of the root-knot nematode M. incognita into tomato roots, but live nematodes had no effect. Infective juvenile M. incognita were repelled from all entomopathogenic nematode treatments that included their symbiotic bacteria. They were repelled by Galleria mellonella cadavers infected with S. carpocapsae, S. feltiae, and S. riobrave and from cell-free culture filtrates of the symbiotic bacteria Xenorhabdus nematophilus, X. bovienii, and Xenorhabdus sp. "R" from the three nematode species, respectively. Cell-free filtrates from all three Xenorhabdus spp. were toxic to M. incognita infective juveniles causing 98-100% mortality at 15% concentration. Cell-free filtrate of Xenorhabdus sp. "R" also reduced the hatch of M. incognita eggs. Application of formulated bacterial cell-free filtrates temporarily suppressed M. incognita penetration into tomato roots in a greenhouse trial. The short-term effects of cell-free bacterial filtrates, namely toxicity and repellency, were almost entirely due to ammonium. These results demonstrate allelopathic interactions between plant-parasitic nematodes, entomopathogenic nematodes and their symbiotic bacteria. The likely role of allelopathy in the suppression of plant-parasitic nematodes by innundative applications of entomopathogenic nematodes is discussed. Allelopathie: Ein moglicher Mechanismus zur Unterdruckung pflanzenparasitarer Nematoden durch insektenpathogene Nematoden - Es wird ein moglicher Mechanismus zur Unterdruckung des pflanzenparasitaren Nematoden Meloidogyne incognita durch insektenpathogene Nematoden beschrieben. Durch Hitze abgetotete insektenpathogene Nematoden Steinernema feltiae und S. riobrave underdruckten das Eindringen des Wurzelgallenalchens M. incognita in Tomatenwurzeln, lebende Nematoden hatten keine Wirkung. Infektionsjuvenile von M. incognita wurden von allen Behandlungen mit insektenpathogenen Nematoden abgestossen, die auch die symbiontischen Bakterien einschlossen. Sie wurden durch die Kadaver von Galleria mellonella abgestossen, die mit S. carpocapsae, S. feltiae und S. riobrave infiziert waren sowie durch zellfreie Kultursubstrate der symbiontischen Bakterien Xenorhabdus nematophilus, X. bovienii und Xenorhabdus sp. "R" aus den drei genannten Nematodenarten. Zellfreie Kultursubstrate von allen drei Xenorhabdus spp. waren giftig fur die Infektionsjuvenilen von M. incognita und verursachten in einer Konzentration von 15% Abtotungsraten von 98-100%. Zellfreie Kultursubstrate von Xenorhabdus sp. "R" vermiderten ausserdem das Schlupfen von M. incognita-Eiern. In einem Gewachshausversuch unterdruckten formulierte zellfreie Bakterienfiltrate vorubergehend das Eindringen von M. incognita in Tomatenwurzeln. Die Kurzzeitwirkungen von zellfreien Bakterien filtraten, namentlich Giftigkeit und Abstossung, waren nahezu ganz bedingt durch Ammoniak. Diese Ergebnisse zeigen das Vorhandensein von allelopathischen Wechselwirkungen zwischen pflanzenparasitaren Nematoden, insektenpathogenen Nematoden und deren symbiontischen Bakterien. Die wahrscheinliche Rolle von Allelopathie bei der Unterdruckung pflanzenparasitarer Nematoden durch eine Massenanwendung insektenpathogener Nematoden wird diskutiert.


2016 ◽  
Vol 283 (1835) ◽  
pp. 20160942 ◽  
Author(s):  
Jinshui Zheng ◽  
Donghai Peng ◽  
Ling Chen ◽  
Hualin Liu ◽  
Feng Chen ◽  
...  

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.


Nematology ◽  
2020 ◽  
pp. 1-17 ◽  
Author(s):  
Tim C. Thoden ◽  
Mariam A. Alkader ◽  
John A. Wiles

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.


2011 ◽  
Vol 48 (2) ◽  
pp. 124-136 ◽  
Author(s):  
V. Čermák ◽  
V. Gaar ◽  
L. Háněl ◽  
K. Široká

AbstractComposition and vertical distribution of soil nematode communities within soil profile were investigated in eight hop gardens in Czech Republic. In total, the presence of 78 nematode genera was confirmed. Genus Drilocephalobus (Coomans & Coomans, 1990) is new for fauna of the Czech Republic. The highest abundance of soil nematodes was found at a depth of 0–10 cm and declined with increasing depth of soil profile. The most dominant genus was Bitylenchus, followed by genera Acrobeloides, Ditylenchus, Chiloplacus and Cervidelus. Ten genera of plant parasitic nematodes were recorded: Bitylenchus (with prevalence of B. dubius), Helicotylenchus, Heterodera (with absolute prevalence of H. humuli), Geocenamus, Longidorella, Longidorus (only L. elongatus), Merlinius (with prevalence of M. brevidens), Paratylenchus and Pratylenchus. Low population densities of predators and omnivores, low values of the community indices (MI, ΣMI, SI, and CI), and high values of NCR, EI, and PPI/MI ratio indicated disturbed nematode communities in hop gardens and bacteria-dominated decomposition pathways in the soil food web.


Plant Disease ◽  
2006 ◽  
Vol 90 (4) ◽  
pp. 471-475 ◽  
Author(s):  
Enrique E. Pérez ◽  
Edwin E. Lewis

A 2-year experiment was conducted to test suppression of plant-parasitic nematodes on English boxwood using entomopathogenic nematodes and 3.5% thyme oil formulated as Promax. Treatments were Steinernema riobrave formulated as BioVector and S. feltiae formulated as Nemasys, both applied at a rate of 2.5 billion infective juveniles/ha, thyme oil at rate of 9.3 liters/ha, and nontreated control. In the 2001 season, treatment with S. feltiae reduced (P ≤ 0.05) the population growth of Tylenchorhynchus sp. 7 days after treatment and Hoplolaimus sp. 30 and 60 days after treatment. Treatment with S. riobrave reduced (P ≤ 0.05) the population growth of all plant-parasitic nematode species at all sampling dates, with the exception of Mesocriconema sp. 30 days after treatment and Tylenchorhynchus sp. and Rotylenchus buxophilus 60 days after treatment. Treatment with thyme oil reduced (P ≤ 0.05) the population growth of all plant-parasitic nematode genera at all sampling dates except Tylenchorhynchus sp. and R. buxophilus 60 days after treatment. In the 2002 season, treatment with S. feltiae had no effect on nematode population growth. Treatment with S. riobrave reduced (P ≤ 0.05) the population growth of R. buxophilus 7 days after treatment, and all plant-parasitic nematodes 30 and 60 days after treatment except Hoplolaimus sp. 30 days after treatment and Mesocriconema sp. 60 days after treatment. Treatment with thyme oil reduced (P ≤ 0.05) the population growth at all sampling dates of plant-parasitic nematodes except Mesocriconema sp. 60 days after treatment.


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