Fig 1.
Characterization of the de novo assembly of the Macrosiphum euphorbiae transcriptome.
Distributions of (A) Sequence length (B) BLASTx E-values and (C) Sequence BLASTx hits similarity.
Fig 2.
The M. euphorbiae transcriptome top hits species distribution.
Data obtained using BLASTx analysis in NCBI’s non-redundant protein database.
Fig 3.
Acyrthosiphon pisum gene coverage by M. euphorbiae contigs.
Histogram showing the coverage of A. pisum predicted genes by the de novo assembled M. euphorbiae transcriptome.
Fig 4.
Histogram of the M. euphorbiae transcriptome gene ontology (GO) classification.
GO level 2 descriptions of the indicated three main categories. The visualization of GO distribution was done with WEGO tool (http://wego.genomics.org.cn/).
Fig 5.
Gene ontology (GO) classification of the endosymbiont Buchnera aphidicola.
GO level 3 description of Buchnera transcripts identified among the M. euphorbiae transcriptome.
Fig 6.
Line up of MeV-2 and MeV-3 sequences with their respective homologous sequences.
(A) Amino acid translation of MeV-2 transcript (GAMO01012456.1; Me_WB16380), with its related virus proteins including: Dysaphis plantaginea DNV (DplDNV; ACI01073.1); Myzus persicae DNV (MpDNV; NP_874375.1); and Acyrthosiphum pisum uncharacterized protein (Ap; XP-016664361.1). (B) Amino acid translation of MeV-3 (GAOM01011582.1; Me_WB14511) with its related virus proteins including: Hubei sobemo-like virus 49 (Hsv49; APG75768.1), Braid Burn Virus (BBv; AMO03213.1), Hubei sobemo-like virus 48 (Hsv48; APG75765.1), La Tardoire virus (LTv; AMO03214.1), and Wuhan insect virus 34 (Wiv34; APG75723.1). Amino acids in red indicate high consensus, blue low consensus and black neutral.
Fig 7.
MeV-2 and MeV3 are present in M. euphorbiae WU11 population and is vertically transmitted to progeny.
Aphid nucleic acids were used in RT-PCR for MeV-2 and MeV-3 detection. For evaluation of vertical transmission, first instar nymphs were collected while being laid from adult aphids, before touching tomato leaflets, using a brush and transferred to a naïve tomato plant. One week later, when nymphs had molted into adults, single aphids were processed for the presence of MeV-2 and MeV-3. Aphid ribosomal gene RpL27 was used as positive control. M = molecular weight marker.
Fig 8.
MeV-2 and MeV3 are present in different M. euphorbiae (Me) populations.
Nucleic acids from mixed developmental stages of aphids were used in RT-PCR for the detection of MeV-2 and MeV-3. Macrosiphum euphorbiae from Germany (DEU), the Netherlands (NDL), France (FR) the United States of America (USA), and Canada (CAN) were used. The population from France is WU11 colony from which the virus was identified. Arabic numerals stand for different aphid populations. FR1a and FR1b colonies are from the same M. euphorbiae population separated for at least 14 years. Aphid ribosomal gene RpL27 was used as positive control. M = molecular weight marker. The cropped two lanes of the MeV-3 gel, displays enhanced imaging of the two amplified bands.
Fig 9.
MeV-2 is delivered by M. euphorbiae (Me) into plant tissues during feeding but does not persist in the plant in the absence of the aphid.
(A) Nucleic acids isolated from leaves of naïve tomato plants or from plants infested for 2 weeks with MeV-2-infected M. euphorbiae were used in RT-PCR. (B, C) M. euphorbiae heavily infested tomato leaves were cleared of the aphids. Leaflets were cut into halves longitudinally through the midrib and the detached half was processed for MeV-2 (B) or MeV-3 (C) detection. The second half of the leaflet was left attached to the plant, free of aphids, for 14 additional days before processing. SlUbi3 was used as a positive control. M = molecular weight marker.