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flp-32 Ligand/receptor silencing phenocopy faster plant pathogenic nematodes.

Atkinson LE, Stevenson M, McCoy CJ, Marks NJ, Fleming C, Zamanian M, Day TA, Kimber MJ, Maule AG, Mousley A - PLoS Pathog. (2013)

Bottom Line: This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate.This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R).This is the first functional characterisation of a parasitic nematode FLP-GPCR.

View Article: PubMed Central - PubMed

Affiliation: Molecular Biosciences-Parasitology, Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast, United Kingdom.

ABSTRACT
Restrictions on nematicide usage underscore the need for novel control strategies for plant pathogenic nematodes such as Globodera pallida (potato cyst nematode) that impose a significant economic burden on plant cultivation activities. The nematode neuropeptide signalling system is an attractive resource for novel control targets as it plays a critical role in sensory and motor functions. The FMRFamide-like peptides (FLPs) form the largest and most diverse family of neuropeptides in invertebrates, and are structurally conserved across nematode species, highlighting the utility of the FLPergic system as a broad-spectrum control target. flp-32 is expressed widely across nematode species. This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate. This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R). This is the first functional characterisation of a parasitic nematode FLP-GPCR.

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Globodera pallida flp-32 (Gp-flp-32) and FLP-32 receptor (Gp-flp-32R) silenced nematodes exhibit accelerated migration rates.siRNA soaking reduces Gp-flp-32 and Gp-flp-32R transcript levels by 55% and 75% respectively (A) compared to controls (control siRNA and untreated worms; P<0.001). Post-RNAi Gp-flp-32 and Gp-flp-32R silenced worms displayed an increased speed of migration down a vertical sand column when compared to control worms (B and C). After 4 h migration 84% of Gp-flp-32 and 91% of Gp-flp-32R worms had migrated compared to 59% and 64% of control siRNA and untreated worms respectively (P<0.01; B and C), after 6 h all Gp-flp-32 and Gp-flp-32R silenced worms had completed migration (B and C). There was no significant difference in the migratory ability of Gp-flp-32 and Gp-flp-32R silenced worms over the 6 h migration period (C).
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ppat-1003169-g004: Globodera pallida flp-32 (Gp-flp-32) and FLP-32 receptor (Gp-flp-32R) silenced nematodes exhibit accelerated migration rates.siRNA soaking reduces Gp-flp-32 and Gp-flp-32R transcript levels by 55% and 75% respectively (A) compared to controls (control siRNA and untreated worms; P<0.001). Post-RNAi Gp-flp-32 and Gp-flp-32R silenced worms displayed an increased speed of migration down a vertical sand column when compared to control worms (B and C). After 4 h migration 84% of Gp-flp-32 and 91% of Gp-flp-32R worms had migrated compared to 59% and 64% of control siRNA and untreated worms respectively (P<0.01; B and C), after 6 h all Gp-flp-32 and Gp-flp-32R silenced worms had completed migration (B and C). There was no significant difference in the migratory ability of Gp-flp-32 and Gp-flp-32R silenced worms over the 6 h migration period (C).

Mentions: Here we used RNAi soaking experiments, measurements of post-silencing changes in Gp-flp-32 transcript levels, and bioassays to assess nematode phenotype, in an attempt to elucidate the role of flp-32 in PCN. Consistent and statistically significant reduction in target transcript (quantified as ΔΔCt of Gp-flp-32 transcript relative to Gp-ace reference transcript) of 55.1±4.6% (n = 3) was achieved in Gp-flp-32 siRNA treated worms when compared to untreated worms (P<0.001, q = 8.988) and non-native control siRNA treated worms (P<0.001, q = 9.716; see Fig. 4A).


flp-32 Ligand/receptor silencing phenocopy faster plant pathogenic nematodes.

Atkinson LE, Stevenson M, McCoy CJ, Marks NJ, Fleming C, Zamanian M, Day TA, Kimber MJ, Maule AG, Mousley A - PLoS Pathog. (2013)

Globodera pallida flp-32 (Gp-flp-32) and FLP-32 receptor (Gp-flp-32R) silenced nematodes exhibit accelerated migration rates.siRNA soaking reduces Gp-flp-32 and Gp-flp-32R transcript levels by 55% and 75% respectively (A) compared to controls (control siRNA and untreated worms; P<0.001). Post-RNAi Gp-flp-32 and Gp-flp-32R silenced worms displayed an increased speed of migration down a vertical sand column when compared to control worms (B and C). After 4 h migration 84% of Gp-flp-32 and 91% of Gp-flp-32R worms had migrated compared to 59% and 64% of control siRNA and untreated worms respectively (P<0.01; B and C), after 6 h all Gp-flp-32 and Gp-flp-32R silenced worms had completed migration (B and C). There was no significant difference in the migratory ability of Gp-flp-32 and Gp-flp-32R silenced worms over the 6 h migration period (C).
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Related In: Results  -  Collection

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ppat-1003169-g004: Globodera pallida flp-32 (Gp-flp-32) and FLP-32 receptor (Gp-flp-32R) silenced nematodes exhibit accelerated migration rates.siRNA soaking reduces Gp-flp-32 and Gp-flp-32R transcript levels by 55% and 75% respectively (A) compared to controls (control siRNA and untreated worms; P<0.001). Post-RNAi Gp-flp-32 and Gp-flp-32R silenced worms displayed an increased speed of migration down a vertical sand column when compared to control worms (B and C). After 4 h migration 84% of Gp-flp-32 and 91% of Gp-flp-32R worms had migrated compared to 59% and 64% of control siRNA and untreated worms respectively (P<0.01; B and C), after 6 h all Gp-flp-32 and Gp-flp-32R silenced worms had completed migration (B and C). There was no significant difference in the migratory ability of Gp-flp-32 and Gp-flp-32R silenced worms over the 6 h migration period (C).
Mentions: Here we used RNAi soaking experiments, measurements of post-silencing changes in Gp-flp-32 transcript levels, and bioassays to assess nematode phenotype, in an attempt to elucidate the role of flp-32 in PCN. Consistent and statistically significant reduction in target transcript (quantified as ΔΔCt of Gp-flp-32 transcript relative to Gp-ace reference transcript) of 55.1±4.6% (n = 3) was achieved in Gp-flp-32 siRNA treated worms when compared to untreated worms (P<0.001, q = 8.988) and non-native control siRNA treated worms (P<0.001, q = 9.716; see Fig. 4A).

Bottom Line: This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate.This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R).This is the first functional characterisation of a parasitic nematode FLP-GPCR.

View Article: PubMed Central - PubMed

Affiliation: Molecular Biosciences-Parasitology, Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast, United Kingdom.

ABSTRACT
Restrictions on nematicide usage underscore the need for novel control strategies for plant pathogenic nematodes such as Globodera pallida (potato cyst nematode) that impose a significant economic burden on plant cultivation activities. The nematode neuropeptide signalling system is an attractive resource for novel control targets as it plays a critical role in sensory and motor functions. The FMRFamide-like peptides (FLPs) form the largest and most diverse family of neuropeptides in invertebrates, and are structurally conserved across nematode species, highlighting the utility of the FLPergic system as a broad-spectrum control target. flp-32 is expressed widely across nematode species. This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate. This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R). This is the first functional characterisation of a parasitic nematode FLP-GPCR.

Show MeSH
Related in: MedlinePlus