Dataset Information

Oral delivery of double-stranded RNAs and siRNAs induces RNAi effects in the potato/tomato psyllid, Bactericerca cockerelli.

ABSTRACT: The potato/tomato psyllid, Bactericerca cockerelli (B. cockerelli), and the Asian citrus psyllid, Diaphorina citri (D. citri), are very important plant pests, but they are also vectors of phloem-limited bacteria that are associated with two devastating plant diseases. B. cockerelli is the vector of Candidatus Liberibacter psyllaurous (solanacearum), which is associated with zebra chip disease of potatoes, and D. citri is the vector of Ca. Liberibacter asiaticus, which is associated with the Huanglongbing (citrus greening) disease that currently threatens the entire Florida citrus industry. Here we used EST sequence information from D. citri to identify potential targets for RNA interference in B. cockerelli. We targeted ubiquitously expressed and gut-abundant mRNAs via injection and oral acquisition of double-stranded RNAs and siRNAs and were able to induce mortality in recipient psyllids. We also showed knockdown of target mRNAs, and that oral acquisition resulted primarily in mRNA knockdown in the psyllid gut. Concurrent with gene knockdown was the accumulation of target specific ? 21 nucleotide siRNAs for an abundant mRNA for BC-Actin. These results showed that RNAi can be a powerful tool for gene function studies in psyllids, and give support for continued efforts for investigating RNAi approaches as possible tools for psyllid and plant disease control.

SUBMITTER: Wuriyanghan H

PROVIDER: S-EPMC3218023 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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Oral delivery of double-stranded RNAs and siRNAs induces RNAi effects in the potato/tomato psyllid, Bactericerca cockerelli.

Wuriyanghan Hada H Rosa Cristina C Falk Bryce W BW

PloS one 20111116 11

The potato/tomato psyllid, Bactericerca cockerelli (B. cockerelli), and the Asian citrus psyllid, Diaphorina citri (D. citri), are very important plant pests, but they are also vectors of phloem-limited bacteria that are associated with two devastating plant diseases. B. cockerelli is the vector of Candidatus Liberibacter psyllaurous (solanacearum), which is associated with zebra chip disease of potatoes, and D. citri is the vector of Ca. Liberibacter asiaticus, which is associated with the Huan ...[more]

PMID: 22110747

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Project description:Purpose: The tomato psyllid, Bactericera cockerelli Šulc (Hemiptera: Triozidae), is a pest of tomato (Solanum lycopersicum) and potato (S. tuberosum) in the U.S. and vectors the disease-causing pathogen ‘Candidatus Liberibacter solanacearum’. Plants undergo physiological, transcriptomic, or epigenetic changes in order to mount a stronger, faster response against secondary challenges by previously perceived threats. This is called defense ‘priming’ and it likely has an impact on vectored disease transmission. Currently, it is still unknown whether or not psyllid infestation has any lasting consequences for tomato gene expression or defense. To characterize the genes potentially involved in tomato priming against psyllids, RNA was extracted from psyllid-primed and uninfested tomato (Moneymaker) leaves three weeks after infestation. Methods: RNA was extracted and sequenced from plants three weeks after psyllid infestation. Plants were either left alone (Control or C) or infested with psyllids (Primed or J1). Libraries were developed using the TruSeq RNA Library Prep Kit v2. Sequencing was performed on the Illumina PE HiSeq 2500 v4 platform. Processed sequences were uploaded to the CyVerse Discovery Environment computational infrastructure where bioinformatic analysis was performed using the Tuxedo Suite 2 workflow. Results: Illumina HiSeq sequencing of tomato cDNA libraries produced 132,428,443 total reads that met FastQC quality control criteria. 94.6% of all reads mapped to vSL3.0 of the S. lycopersicum genome. CuffDiff2 analysis identified 310 differentially expressed genes (DEGs) between control and psyllid-primed plants (q-value <0.01). Conclusions: A week-long infestation by a small number of B. cockerelli had lasting consequences for gene expression in tomato plants. Homologs of the DEGs were associated with 1) defense against abiotic and biotic stress, 2) growth and development, and 3) components of plant biology indirectly involved in plant growth and development such as homeostasis, transcription/translation, and molecular transport.

Project description:In Central and North America, Australia and New Zealand, potato (Solanum tuberosum) crops are attacked by Bactericera cockerelli, the tomato potato psyllid (TPP). 'Mesh crop covers' which are used in Europe and Israel to protect crops from insect pests, have been used experimentally in New Zealand for TPP control. While the covers have been effective for TPP management, the green peach aphid (GPA, Myzus persicae) has been found in large numbers under the mesh crop covers. This study investigated the ability of the GPA to penetrate different mesh hole sizes. Experiments using four sizes (0.15 × 0.15, 0.15 × 0.35, 0.3 × 0.3 and 0.6 × 0.6 mm) were carried out under laboratory conditions to investigate: (i) which mesh hole size provided the most effective barrier to GPA; (ii) which morph of adult aphids (apterous or alate) and/or their progeny could breach the mesh crop cover; (iii) would leaves touching the underside of the cover, as opposed to having a gap between leaf and the mesh, increase the number of aphids breaching the mesh; and (iv) could adults feed on leaves touching the cover by putting only their heads and/or stylets through it? No adult aphids, either alate or apterous, penetrated the mesh crop cover; only nymphs did this, the majority being the progeny of alate adults. Nymphs of the smaller alatae aphids penetrated the three coarsest mesh sizes; nymphs of the larger apterae penetrated the two coarsest sizes, but no nymphs penetrated the smallest mesh size. There was no statistical difference in the number of aphids breaching the mesh crop cover when the leaflets touched its underside compared to when there was a gap between leaf and mesh crop cover. Adults did not feed through the mesh crop cover, though they may have been able to sense the potato leaflet using visual and/or olfactory cues and produce nymphs as a result. As these covers are highly effective for managing TPP on field potatoes, modifications of this protocol are required to make it effective against aphids as well as TPP.

Dataset Information

Oral delivery of double-stranded RNAs and siRNAs induces RNAi effects in the potato/tomato psyllid, Bactericerca cockerelli.

Publications

Oral delivery of double-stranded RNAs and siRNAs induces RNAi effects in the potato/tomato psyllid, Bactericerca cockerelli.

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