Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

MRNA time-course transcription profiling of bacteriocytes and gut tissue of pea aphids reared on standard artificial diet (AP3) and artificial diet depleted in Tyr and Phe (YFÃ)

ABSTRACT: Tyrosine (Tyr, Y) and phenylalanine (Phe, F) synthesis is shared by the pea aphid and its symbiont Buchnera aphidicola.These aromatic amino acids are essential for the pea aphid growth and development. To characterize the molecular mechanisms, at gene transcriptional level, underlying this symbiotic integrated network pea aphids (Acyrthosyphon pisum, clone LL01) were reared on (i) standard artificial diet (AP3) and (ii) on the same AP3 medium depleted of Tyr (Y) and Phe (F). From each of the two groups, aphids were collected at specific time points and dissected: 12 h (D0), 1 day (D1), 2 days (D2), 3 days (D3), 4 days (D4), 5 days (D5) and 7 days (D7). Total RNA, to be used in gene expression analysis by arrays, was extracted, under the two rearing conditions, from two tissues: gut [from 20 aphids per sample at all 7 time points] and bacteriocytes [from 25 aphids per sample at 4 time points: 3 days (D3), 4 days (D4), 5 days (D5) and 7 days (D7)]. At each time point we included three biological replicates.

ORGANISM(S): Acyrthosiphon pisum

SUBMITTER: Stefano Colella

PROVIDER: E-MTAB-4456 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Aphids, sap-sucking insects in the order Hemiptera, are among the most prolific insect vectors of plant viruses Plant viruses from the family Luteoviridae are transmitted exclusively by aphids in a circulative manner and cause significant crop yield losses. Circulative plant viruses must cross the aphid gut and other tissues prior to transmission to a new host plant. The discovery of proteins that control acquisition and transmission in the insect vector is the biggest challenge for the vector biology field and will have practical applications for growers by providing new molecular targets for the development of precision vector management tools. The green peach aphid, Myzus persicae, is a vector of the Potato leafroll virus (PLRV), a polerovirus in the Luteoviridae. PLRV transmission efficiency was significantly reduced when a clonal lineage of M. persicae was reared on turnip (T-Myzus) as compared to the weed physalis (P-Myzus). The effect on PLRV transmission efficiency was transient and caused by a host-switch response. Using 2-D DIGE, we revealed that the major difference in the proteome profile of P- and T-Myzus was the lysosomal cysteine protease cathepsin B, with multiple size and charge isoforms of this enzyme up-regulated in T-Myzus. Quantitative, shotgun proteomics revealed a specific upregulation in the expression of other lysosomal proteins in T-Myzus as compared to P-Myzus, including cathepsin B, cathepsin B-16, beta-glucuronidase, peroxidasin, legumain-like, and aminopeptidase-N. The titer of PLRV was over 1.5 fold higher in P-Myzus than in T-Myzus at 24h and 72h after the beginning of virus acquisition, suggesting that virus acquisition in P-Myzus was more efficient. Cathepsin B and PLRV localization were starkly different in P- and T-Myzus midguts, the site of PLRV acquisition into the insect. In P-Myzus midguts, an abundance of PLRV was observed inside midgut cells, and cathepsin B was sequestered in a subcellular compartment. In contrast, there is near complete co-localization of cathepsin B and PLRV at the cell membranes in viruliferous T-Myzus. Inhibition of cathepsin and other cysteine proteases with E64 restored the ability of T-Myzus to transmit PLRV in a dose-dependent manner, suggesting that the activities of lysosomal cysteine proteases at the cell membrane in T-Myzus is responsible for the change in virus transmission phenotype in these aphids. T-Myzus individuals weighed more and had more progeny than P-Myzus individuals. These data are all consistent with the hypothesis that there is an induction of lysosomal exocytos in the midgut of T-Myzus linked to the ability of the aphid to acquire PLRV. These data also show that the ability of the generalist aphid M. persicae to transmit PLRV is influenced by the host plant the aphids are reared on, information that is useful to growers for polerovirus management in field crops.

Project description:Compatibility between aphids and plant hosts is genetically determined by both interactingorganisms. For example, plants may carry resistance (R) genes or deploy chemical defences.Aphid saliva contains many proteins that are secreted into host tissues. Subsets of these proteins are predicted to act as effectors, either subverting or triggering host immunity. However, associating particular effectors with virulence or avirulence outcomes presents challenges due to the combinatorial complexity. Here we use defined aphid and host geneticsto test for co-segregation of expressed aphid transcripts and proteins with virulent or avirulent phenotypes. We compared virulent and avirulent pea aphid parental genotypes, and their bulk segregant F1 progeny on Medicago truncatula genotypes carrying or lacking the RAP1 resistance quantitative trait locus. Differential expression analysis based on RNA sequencing of whole bodyand head samples, in combination with proteomics of saliva and salivary glands, enabled us to pinpoint proteins associated with virulence/avirulence phenotypes. There was relatively little impact of host genotype, whereas large numbers of transcripts and proteins were differentially expressed between parental aphids, likely a reflection of their classification as divergent biotypes within the pea aphid species complex. Many fewer transcripts intersected with the equivalent differential expression patterns in the bulked F1F1 progeny, providing an effective filter for removing genomic background effects. Overall, there were more upregulated genes detected in the F1 avirulent dataset compared with the virulent one. Some of the differentially expressed transcripts were also found in the differentially expressed proteomes, with aminopeptidase N proteins being the most frequent differentiallyexpressed family. In addition, a substantial proportion (23%) of salivary proteins lack annotations, suggesting that many novel functions remain to be discovered.

Dataset Information

MRNA time-course transcription profiling of bacteriocytes and gut tissue of pea aphids reared on standard artificial diet (AP3) and artificial diet depleted in Tyr and Phe (YFÃ)

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Dataset Information

MRNA time-course transcription profiling of bacteriocytes and gut tissue of pea aphids reared on standard artificial diet (AP3) and artificial diet depleted in Tyr and Phe (YFÃ)

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

MRNA time-course transcription profiling of bacteriocytes and gut tissue of pea aphids reared on standard artificial diet (AP3) and artificial diet depleted in Tyr and Phe (YFÃ)