Immunolocalization of a putative cuticular collagen protein in several developmental stages of Meloidogynearenaria, Globodera pallida and G. rostochiensis

2002 ◽  
Vol 76 (1) ◽  
pp. 1-6 ◽  
Author(s):  
I.M. de O. Abrantes ◽  
R.H.C. Curtis
Keyword(s):  
Developmental Stages ◽  
Globodera Pallida ◽  
Parasitic Nematodes ◽  
Cdna Expression Library ◽  
In Planta ◽  
Cyst Nematodes ◽  
Expression Library ◽  
Second Stage ◽  
Collagen Protein ◽  
Cuticular Surface

AbstractThe monoclonal antibody IACR-CCNj.3d has previously been used to isolate a gene (gp-col-8) with strong similarity to cuticular collagen from a mixed stage Globodera pallida cDNA expression library. The antibody has also been shown to label specifically the amphidial canal of pre-parasitic second stage juveniles (J2) of several plant nematode species without any reactivity on the cuticular surface, indicating that this protein is either not present or is inaccessible on the cuticular surface. This paper investigates the cross-reactivity of Mab IACR-CCNj.3d with Meloidogyne arenaria and the localization of the putative collagen protein on the cuticular surface of parasitic stages in planta and on the cuticular surface of juveniles inside eggs. The antigen was shown to be present in all developmental stages of the two species of potato cyst nematodes and M. arenaria. The antibody bound strongly to the amphidial canal and hypodermis of pre-parasitic J2 and adult females. The antigen was present on the cuticular surface of the sausage-shaped J2 in planta and of first stage juveniles (J1) inside the eggs. The presence of collagen on the surface of the cuticle of moulting stages of plant parasitic nematodes has been observed for the first time. It is clear that this protein has a role in the construction of the cuticle of the first stage juveniles and parasitic second stage juveniles, during moulting inside the eggs and in the root tissue, respectively.

Isolation and localisation of an annexin gene (gp-nex) from the potato cyst nematode, Globodera pallida

Nematology ◽  
2001 ◽  
Vol 3 (1) ◽  
pp. 45-54 ◽  
Author(s):  
Andrew Warry ◽  
Rosane Curtis ◽  
Andrew Porter ◽  
Luca Fioretti ◽  
Patrick Haydock
Keyword(s):  
Amino Acid ◽  
Nematode Species ◽  
Globodera Pallida ◽  
Predicted Amino Acid Sequence ◽  
Cyst Nematodes ◽  
Expression Library ◽  
Calcium Dependent ◽  
Secretion Signal ◽  
Second Stage ◽  
Annexin Gene

AbstractA full length cDNA clone was isolated by screening a mixed stage Globodera pallida expression library using the monospecific polyclonal antibody IACR-PC320. Sequence comparison indicated that the predicted 332 amino acid protein encoded by the cDNA was a member of the annexin gene family named gp-nex and very similar to annexins found in other nematode species. Annexins are calcium-dependent phospholipid binding proteins characterised by four repeated domains approximately 70 amino acids in length. The predicted amino acid sequence of the protein did not contain an N-terminal secretion signal peptide; however, the protein was shown to be present in excretory/secretory products from G. pallida second stage juveniles treated with the neurotransmitter 5 methoxy-N, N dimethyl tryptamine. Gp-nex encodes a protein of 35 kDa and was immunolocalised in the amphids, genital primordium and in the constraining muscles above and below the metacorpus pump chamber of G. pallida second stage juveniles. It is also present in eggs and adult females of the two species of the potato cyst nematodes.

scholarly journals Activation of hatching in diapaused and quiescent Globodera pallida

Parasitology ◽  
2012 ◽  
Vol 140 (4) ◽  
pp. 445-454 ◽  
Author(s):  
JUAN E. PALOMARES-RIUS ◽  
JOHN T. JONES ◽  
PETER J. COCK ◽  
PABLO CASTILLO ◽  
VIVIAN C. BLOK
Keyword(s):  
Bursaphelenchus Xylophilus ◽  
Globodera Pallida ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Cyst Nematodes ◽  
Term Survival ◽  
C Elegans ◽  
Dauer Formation ◽  
Dorsal Gland

SUMMARYThe potato cyst nematodes (PCN) Globodera pallida and G. rostochiensis are major pests of potatoes. The G. pallida (and G. rostochiensis) life cycle includes both diapause and quiescent stages. Nematodes in dormancy (diapause or quiescent) are adapted for long-term survival and are more resistant to nematicides. This study analysed the mechanisms underlying diapause and quiescence. The effects of several compounds (8Br-cGMP, oxotremorine and atropine) on the activation of hatching were studied. The measurements of some morphometric parameters in diapaused and quiescent eggs after exposure to PRD revealed differences in dorsal gland length, subventral gland length and dorsal gland nucleolus. In addition, the expression of 2 effectors (IVg9 and cellulase) was not induced in diapaused eggs in water or PRD, while expression was slightly induced in quiescent eggs. Finally, we performed a comparative study to identify orthologues of C. elegans diapause related genes in plant-parasitic nematodes (G. pallida, Meloidogyne incognita, M. hapla and Bursaphelenchus xylophilus). This analysis suggested that it was not possible to identify G. pallida orthologues of the majority of C. elegans genes involved in the control of dauer formation. All these data suggest that G. pallida may use different mechanisms to C. elegans in regulating the survival stage.

The effects of α-solanine and zinc sulphate on the behaviour of potato cyst nematodes Globodera rostochiensis and G. pallida

Nematology ◽  
2015 ◽  
Vol 17 (9) ◽  
pp. 1105-1111 ◽  
Author(s):  
Vincas Būda ◽  
Rasa Čepulytė-Rakauskienė
Keyword(s):  
Host Plants ◽  
Zinc Sulphate ◽  
Globodera Rostochiensis ◽  
Globodera Pallida ◽  
The Other ◽  
Potato Cyst Nematodes ◽  
Water Control ◽  
Cyst Nematodes ◽  
Second Stage ◽  
Testing Period

Two behavioural assays were carried out: one on the attraction of potato cyst nematodes (PCN), Globodera rostochiensis and G. pallida, to solanaceae-specific secondary metabolite α-solanine, the other on the effect of ZnSO4 (a compound known to suppress chemoreceptors). The first assay demonstrated that α-solanine was attractive to second-stage juveniles (J2) of PCN; G. pallida was attracted to concentrations of 10−4 M and 10−5 M, whereas G. rostochiensis was attracted to 10−5 M. Globodera pallida reacted faster than G. rostochiensis to the same concentration of α-solanine. As α-solanine is produced by host plants of PCN, this compound is attributed to kairomones. The response to α-solanine of nematodes pre-exposed to a 3 mM ZnSO4 solution was significantly suppressed compared to that of water control. The effect was observed throughout the whole testing period (30 min). This is the first evidence that both α-solanine and ZnSO4 can affect the behaviour of hatched J2 of PCN.

Identification and characterization of a cDNA clone-encoding antigen of Eimeria acervulina

Parasitology ◽  
2012 ◽  
Vol 139 (13) ◽  
pp. 1711-1719 ◽  
Author(s):  
HUILI ZHU ◽  
LIXIN XU ◽  
RUOFENG YAN ◽  
XIAOKAI SONG ◽  
FANG TANG ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Cdna Expression Library ◽  
Immunofluorescence Analysis ◽  
Expression Library ◽  
E Coli ◽  
Causative Agents ◽  
The Mean ◽  
Identification And Characterization ◽  
Average Body

SUMMARYEimeria spp. are the causative agents of coccidiosis, a major disease affecting the poultry industry. So far, only a few antigen genes of E. acervulina have been reported. In this study, a clone, named as cSZ-JN2, was identified from a cDNA expression library prepared from E. acervulina sporozoite stage with the ability to stimulate the chicken immune response. The sequence analysis showed that the open reading fragment (ORF) of cSZ-JN2 was 153 bp in size and encoded a predicted protein of 50 amino acids of Mr 5·3 kDa. BLASTN search revealed that cSZ-JN2 had no significant homology with the known genes of E. acervulina or any other organism (GenBank). The recombinant cSZ-JN2 antigen expressed in E. coli was recognized strongly by serum from chickens experimentally infected with E. acervulina. Immunofluorescence analysis using antibody against recombinant cSZ-JN2 indicated that this protein was expressed in sporozoite and merozoite developmental stages. Animal challenge experiments demonstrated that the recombinant protein of cSZ-JN2 and DNA vaccine carrying cSZ-JN2 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens and presented anti-coccidial indices of more than 165. All the above results suggested that the cSZ-JN2 was a novel E. acervulina antigen and could be an effective candidate for the development of a new vaccine against E. acervulina infection.

scholarly journals Quarantine nematode species and pathotypes potentially dangerous for domestic potato production: populations diversity and the genetics of potato resistance

2020 ◽  
Vol 24 (7) ◽  
pp. 705-721
Author(s):  
N. V. Mironenko ◽  
T. A. Gavrilenko ◽  
A. V. Khiutti ◽  
O. S. Afanasenko
Keyword(s):  
Molecular Markers ◽  
Genetic Variability ◽  
Functional Organization ◽  
Potato Production ◽  
Potato Cultivars ◽  
Globodera Pallida ◽  
Parasitic Nematodes ◽  
Cyst Nematodes ◽  
Resistant Cultivars ◽  
Potato Resistance

The review considers quarantine species and nematode pathotypes potentially dangerous for domestic potato production. Potatoes are affected by more than 30 types of parasitic nematodes, but the review focuses on the most harmful representatives of genera that cause great damage to potato production: Globodera, Ditylenchus, Nacob bus and Meloidogyne. Phytopathological and molecular methods of identification of species and pathotypes and the main achievements in studying the population variability of parasitic potato nematodes were analyzed. It was shown that due to the peculiarities of the life cycle of nematodes and lability of their genomes, the genetic variability of these organisms is very high, which creates a threat of forming new pathogenic genotypes of the parasites. The information about the intra- and interpopulation variability of nematodes is important for studying the ways of introduction and distribution of separate species, as well as for searching for the correlations of molecular markers with the pathotype. Phylogenetic studies based on modern data on genetic variability of populations have allowed to reveal species complexes in Globodera pallida (Stone) Behrens and Nacobbus aberrans (Thorne) Thorne & Allen (sensu lato), including cryptic species. The main components of successful protection preventing a wide distribution of parasitic nematodes are quarantine measures, agricultural techniques, biological methods of protection and cultivation of resistant cultivars. Special attention in the review is paid to the breeding of potato cultivars with durable resistance to various nematode pathotypes, because the cultivation of such varieties is the most ecologically safe and economically advantageous way to prevent epiphytoties. Currently, significant progress has been made in the genetic protection of potato cultivars, especially against cyst-forming nematodes. The review provides data on sources of potato resistance to parasitic nematodes identified in collections of wild and cultivated species. Data on identified R-gens and QTL of resistance that have been introduced into breeding varieties using different methods and approaches are analyzed. The literature data on the study of structural and functional organization of genes for resistance to potato cyst nematodes are given. The results of molecular research on revealing the polymorphisms of loci involved in the control of resistance to cyst and gall nematodes, the development of molecular markers of certain genes and their use in marker-assisted selection for developing of new resistant cultivars, including those with group resistance, are considered.

scholarly journals Assessment of the nematicidal potential of vermicompost, vermicompost tea, and urea application on the potato-cyst nematodes Globodera rostochiensis and Globodera pallida

2015 ◽  
Vol 55 (2) ◽  
pp. 187-192 ◽  
Author(s):  
Marek Renčo ◽  
Peter Kováčik
Keyword(s):  
Aqueous Solutions ◽  
Growth Parameters ◽  
Control Plant ◽  
Globodera Rostochiensis ◽  
Globodera Pallida ◽  
Parasitic Nematodes ◽  
Potato Cyst Nematodes ◽  
Cyst Nematodes ◽  
Positive Effects ◽  
Vermicompost Tea

Abstract The addition of organic material to the soil can be an effective alternative to the environmentally unsafe chemical treatments that are used to control plant parasitic nematodes. We evaluated the effects of vermicompost alone, and aqueous solutions of vermicompost (vermicompost tea) either alone or mixed with urea, on the development and survival of two potato-cyst nematodes: Globodera rostochiensis (pathotype Ro1) and G. pallida (pathotype Pa2) and on the growth parameters of the host potato plants. Soil amendments with these materials significantly decreased the number of cysts · 400 g–1 of both species in the soil, the number of eggs and juveniles · cyst–1 of both species, and the number of eggs and juveniles · g–1 of both species in the soil, relative to the untreated controls. The suppressive effect was significantly higher at the highest dose than the lowest treatment dose, for all tested materials. Globodera rostochiensis was more sensitive to all the tested materials than G. pallida. The aqueous solutions of vermicompost alone or in combination with urea were more effective than the solid vermicompost used alone, for controlling both species. Vermicompost and the vermicompost teas had positive effects on plant fresh stem weight and stem height. The application of vermicompost tea instead of the solid vermicompost, substantially decreased the amount of material needed. These amendments are thus promising for the control of potato-cyst nematodes in sustainable agricultural systems.

scholarly journals SIZ1 is a nuclear host target of the nematode effector GpRbp1 from Globodera pallida that acts as a negative regulator of basal plant defense to cyst nematodes

2019 ◽  
Author(s):  
Amalia Diaz-Granados ◽  
Mark G. Sterken ◽  
Jarno Persoon ◽  
Hein Overmars ◽  
Somnath S. Pokhare ◽  
...  
Keyword(s):  
Plant Morphology ◽  
Globodera Pallida ◽  
Negative Regulator ◽  
Host Cells ◽  
In Planta ◽  
Cyst Nematodes ◽  
Interaction Sites ◽  
Successful Infection ◽  
Pathogen Effector ◽  
Plant Nematode

AbstractSoil-borne cyst nematodes are obligatory sedentary parasites that cause severe losses to cultivation of major crops such as potato and soybean. Cyst nematodes establish specialised permanent feeding sites within the roots of their host by manipulating plant morphology and physiology through secreted effectors. Here we identified host targets of effector GpRbp-1 and studied their roles in plant-nematode interactions. GpRbp-1 was found to interact in yeast and in planta with the potato and Arabidopsis homologues of Siz/PIAS-type E3 SUMO ligase SIZ1. Our results show that a pathogen effector targets the master regulator SIZ1 in plant cells, which has not been demonstrated earlier to our knowledge. The interaction of GpRbp-1 and SIZ1 localizes to the plant nucleus, suggesting that the nuclear functions of SIZ1 as regulator of plant immunity and physiology may be modulated by GpRbp-1. Furthermore, nematode infection assays and transcriptomic profiling indicate that SIZ1 is required for susceptibility to cyst nematodes. So, these data indicate that E3 SUMO ligases may play an important role in plant-nematode interactions. Based on the prediction of SUMO acceptor and interaction sites in GpRbp-1, a model is proposed in which the effector may recruit SIZ1 to be SUMOylated for full functionality in host cells.Significance statementHere we show that a pathogen effector can target SIZ1, a key protein involved in regulating several aspects of plant biology, most likely to manipulate the SUMOylation of host proteins for successful infection of plants.

scholarly journals Members of the Meloidogyne Avirulence Protein Family Contain Multiple Plant Ligand-Like Motifs

2014 ◽  
Vol 104 (8) ◽  
pp. 879-885 ◽  
Author(s):  
William B. Rutter ◽  
Tarek Hewezi ◽  
Tom R. Maier ◽  
Melissa G. Mitchum ◽  
Eric L. Davis ◽  
...  
Keyword(s):  
Host Plants ◽  
Effector Proteins ◽  
Parasitic Nematodes ◽  
In Planta ◽  
Root Knot Nematode ◽  
Cyst Nematodes ◽  
Root Knot Nematodes ◽  
Cle Peptides ◽  
Variable Domains ◽  
Meloidogyne Spp

Sedentary plant-parasitic nematodes engage in complex interactions with their host plants by secreting effector proteins. Some effectors of both root-knot nematodes (Meloidogyne spp.) and cyst nematodes (Heterodera and Globodera spp.) mimic plant ligand proteins. Most prominently, cyst nematodes secrete effectors that mimic plant CLAVATA3/ESR-related (CLE) ligand proteins. However, only cyst nematodes have been shown to secrete such effectors and to utilize CLE ligand mimicry in their interactions with host plants. Here, we document the presence of ligand-like motifs in bona fide root-knot nematode effectors that are most similar to CLE peptides from plants and cyst nematodes. We have identified multiple tandem CLE-like motifs conserved within the previously identified Meloidogyne avirulence protein (MAP) family that are secreted from root-knot nematodes and have been shown to function in planta. By searching all 12 MAP family members from multiple Meloidogyne spp., we identified 43 repetitive CLE-like motifs composing 14 unique variants. At least one CLE-like motif was conserved in each MAP family member. Furthermore, we documented the presence of other conserved sequences that resemble the variable domains described in Heterodera and Globodera CLE effectors. These findings document that root-knot nematodes appear to use CLE ligand mimicry and point toward a common host node targeted by two evolutionarily diverse groups of nematodes. As a consequence, it is likely that CLE signaling pathways are important in other phytonematode pathosystems as well.

scholarly journals Ingestion of Double-Stranded RNA by Preparasitic Juvenile Cyst Nematodes Leads to RNA Interference

2002 ◽  
Vol 15 (8) ◽  
pp. 747-752 ◽  
Author(s):  
P. E. Urwin ◽  
Catherine J. Lilley ◽  
Howard J. Atkinson
Keyword(s):  
Rna Interference ◽  
Globodera Pallida ◽  
Northern Analysis ◽  
Parasite Development ◽  
Parasitic Nematodes ◽  
Normal Female ◽  
Plant Parasitic Nematodes ◽  
Cyst Nematodes ◽  
Double Stranded Rna ◽  
Plant Parasitic

RNA interference is of value in determining gene function in many organisms. Plant parasitic nematodes are refractory to microinjection as a means of introducing RNA and do not show any oral uptake until they are within plants. We have used octopamine to stimulate uptake by prepara-sitic second stage juveniles of two cyst nematodes, Heterodera glycines and Globodera pallida. This new technique was used to facilitate uptake of double stranded RNA (dsRNA) together with fluoroscein isothiocyanate as a visual marker. Targeting cysteine proteinases did not reduce the number of parasites but caused a shift from the normal female/male ratio of 3:1 to 1:1 by 14 days postinfection (dpi). Exposure of H. glycines to dsRNA corresponding to a newly characterized protein with homology to C-type lectins did not affect sexual fate, but 41% fewer parasites were recovered from the plants. As expected, treatment with dsRNA corresponding to the major sperm protein (MSP) had no effect on either parasite development or sexual fate over 14 days. Northern analysis showed lower transcript abundance for the two targeted mRNAs that occur in J2, plus a later inhibition for MSP transcripts when males developed sperm at 15 dpi. These findings establish a procedure for RNAi of plant parasitic nematodes.

scholarly journals Gene expression changes in diapause or quiescent potato cyst nematode,Globodera pallida, eggs after hydration or exposure to tomato root diffusate

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e1654 ◽  
Author(s):  
Juan Emilio Palomares-Rius ◽  
Pete Hedley ◽  
Peter J.A. Cock ◽  
Jenny A. Morris ◽  
John T. Jones ◽  
...  
Keyword(s):  
Gene Expression ◽  
Ion Transport ◽  
Globodera Pallida ◽  
Parasitic Nematodes ◽  
Cyst Nematodes ◽  
Microarray Gene Expression ◽  
Tomato Root ◽  
Term Survival ◽  
Microarray Gene Expression Analysis

Plant-parasitic nematodes (PPN) need to be adapted to survive in the absence of a suitable host or in hostile environmental conditions. Various forms of developmental arrest including hatching inhibition and dauer stages are used by PPN in order to survive these conditions and spread to other areas. Potato cyst nematodes (PCN) (Globodera pallidaandG. rostochiensis) are frequently in an anhydrobiotic state, with unhatched nematode persisting for extended periods of time inside the cyst in the absence of the host. This paper shows fundamental changes in the response of quiescent and diapaused eggs ofG. pallidato hydration and following exposure to tomato root diffusate (RD) using microarray gene expression analysis encompassing a broad set of genes. For the quiescent eggs, 547 genes showed differential expression following hydration vs. hydratation and RD (H-RD) treatment whereas 708 genes showed differential regulation for the diapaused eggs following these treatments. The comparison between hydrated quiescent and diapaused eggs showed marked differences, with 2,380 genes that were differentially regulated compared with 987 genes following H-RD. Hydrated quiescent and diapaused eggs were markedly different indicating differences in adaptation for long-term survival. Transport activity is highly up-regulated following H-RD and few genes were coincident between both kinds of eggs. With the quiescent eggs, the majority of genes were related to ion transport (mainly sodium), while the diapaused eggs showed a major diversity of transporters (amino acid transport, ion transport, acetylcholine or other molecules).

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