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Transcriptome analysis and systemic RNAi response in the African sweetpotato weevil (Cylas puncticollis, Coleoptera, Brentidae).

Prentice K, Pertry I, Christiaens O, Bauters L, Bailey A, Niblett C, Ghislain M, Gheysen G, Smagghe G - PLoS ONE (2015)

Bottom Line: Subsequently, a transcriptome search showed that the necessary RNAi components relevant to the three major RNAi pathways, were found to be expressed in SPW.The body of treated insects showed inhibition of sclerotization, leading eventually to death.Quantitative Real Time PCR (qPCR) confirmed this phenotype to be the result of gene silencing.

View Article: PubMed Central - PubMed

Affiliation: Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, B-9000 Ghent, Belgium; Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, B-9000 Ghent, Belgium; International Potato Center (CIP), Genomics and Biotechnology Program, Nairobi 00603, Kenya.

ABSTRACT
The African sweetpotato weevil (SPW) Cylas puncticollis Boheman is one of the most important constraints of sweetpotato production in Sub-Saharan Africa and yet is largely an uncharacterized insect pest. Here, we report on the transcriptome analysis of SPW generated using an Illumina platform. More than 213 million sequencing reads were obtained and assembled into 89,599 contigs. This assembly was followed by a gene ontology annotation. Subsequently, a transcriptome search showed that the necessary RNAi components relevant to the three major RNAi pathways, were found to be expressed in SPW. To address the functionality of the RNAi mechanism in this species, dsRNA was injected into second instar larvae targeting laccase2, a gene which encodes an enzyme involved in the sclerotization of insect exoskeleton. The body of treated insects showed inhibition of sclerotization, leading eventually to death. Quantitative Real Time PCR (qPCR) confirmed this phenotype to be the result of gene silencing. Together, our results provide valuable sequence data on this important insect pest and demonstrate that a functional RNAi pathway with a strong and systemic effect is present in SPW and can further be explored as a new strategy for controlling this important pest.

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Related in: MedlinePlus

Sequence comparison to other insect genera from the distribution of BLASTX hit (bitscore >50) against the nr protein database of the National Center for Biotechnology Information.
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pone.0115336.g001: Sequence comparison to other insect genera from the distribution of BLASTX hit (bitscore >50) against the nr protein database of the National Center for Biotechnology Information.

Mentions: For BLAST annotation, contigs were first searched for similar insect protein sequences using BLASTX against the non-redundant (nr) protein NCBI database, This BLASTX analysis produced 44,824 hits, representing 50.0% of total contigs (Fig. 1). The number of non-significant hits (50.0%) indicates that the C. puncticollis transcriptome contains unknown sequences that are not yet described in the insect protein sequences databases. For those sequences with a significant match, 87.68% of the contigs are most similar to sequences from coleopteran species: 40.31% to the red flour beetle Tribolium castaneum, which is a worldwide pest of stored food products, 36.51% to the mountain pine beetle Dendroctonus ponderosae sequences, which is a serious forest pest [32] and 10.87% to the Asian long-horn beetle Anoplophora glabripennis sequences, also found to be destructive of forest trees [33]. The remaining 12.32% of all contigs were more similar to the hemipterans Acyrthosiphon pisum (1.78%) and Triatoma infestans (0.94%), the hymenopterans Camponotus floridanus (0.78%) and Cerapachys biroi (0.60%), the dipteran Corathrella appendiculata (0.65%), the lepidopteran Bombyx mori (0.57%) and others (7.1%).


Transcriptome analysis and systemic RNAi response in the African sweetpotato weevil (Cylas puncticollis, Coleoptera, Brentidae).

Prentice K, Pertry I, Christiaens O, Bauters L, Bailey A, Niblett C, Ghislain M, Gheysen G, Smagghe G - PLoS ONE (2015)

Sequence comparison to other insect genera from the distribution of BLASTX hit (bitscore >50) against the nr protein database of the National Center for Biotechnology Information.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4295849&req=5

pone.0115336.g001: Sequence comparison to other insect genera from the distribution of BLASTX hit (bitscore >50) against the nr protein database of the National Center for Biotechnology Information.
Mentions: For BLAST annotation, contigs were first searched for similar insect protein sequences using BLASTX against the non-redundant (nr) protein NCBI database, This BLASTX analysis produced 44,824 hits, representing 50.0% of total contigs (Fig. 1). The number of non-significant hits (50.0%) indicates that the C. puncticollis transcriptome contains unknown sequences that are not yet described in the insect protein sequences databases. For those sequences with a significant match, 87.68% of the contigs are most similar to sequences from coleopteran species: 40.31% to the red flour beetle Tribolium castaneum, which is a worldwide pest of stored food products, 36.51% to the mountain pine beetle Dendroctonus ponderosae sequences, which is a serious forest pest [32] and 10.87% to the Asian long-horn beetle Anoplophora glabripennis sequences, also found to be destructive of forest trees [33]. The remaining 12.32% of all contigs were more similar to the hemipterans Acyrthosiphon pisum (1.78%) and Triatoma infestans (0.94%), the hymenopterans Camponotus floridanus (0.78%) and Cerapachys biroi (0.60%), the dipteran Corathrella appendiculata (0.65%), the lepidopteran Bombyx mori (0.57%) and others (7.1%).

Bottom Line: Subsequently, a transcriptome search showed that the necessary RNAi components relevant to the three major RNAi pathways, were found to be expressed in SPW.The body of treated insects showed inhibition of sclerotization, leading eventually to death.Quantitative Real Time PCR (qPCR) confirmed this phenotype to be the result of gene silencing.

View Article: PubMed Central - PubMed

Affiliation: Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, B-9000 Ghent, Belgium; Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, B-9000 Ghent, Belgium; International Potato Center (CIP), Genomics and Biotechnology Program, Nairobi 00603, Kenya.

ABSTRACT
The African sweetpotato weevil (SPW) Cylas puncticollis Boheman is one of the most important constraints of sweetpotato production in Sub-Saharan Africa and yet is largely an uncharacterized insect pest. Here, we report on the transcriptome analysis of SPW generated using an Illumina platform. More than 213 million sequencing reads were obtained and assembled into 89,599 contigs. This assembly was followed by a gene ontology annotation. Subsequently, a transcriptome search showed that the necessary RNAi components relevant to the three major RNAi pathways, were found to be expressed in SPW. To address the functionality of the RNAi mechanism in this species, dsRNA was injected into second instar larvae targeting laccase2, a gene which encodes an enzyme involved in the sclerotization of insect exoskeleton. The body of treated insects showed inhibition of sclerotization, leading eventually to death. Quantitative Real Time PCR (qPCR) confirmed this phenotype to be the result of gene silencing. Together, our results provide valuable sequence data on this important insect pest and demonstrate that a functional RNAi pathway with a strong and systemic effect is present in SPW and can further be explored as a new strategy for controlling this important pest.

Show MeSH
Related in: MedlinePlus