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On the other hand, the Favret and C1 primer pairs performed well in both monoplex and triplex RT-PCR formats using single aphids of M. persicase, M. euphorbiae and A. nasturtii, demonstrating their suitability to indicate the successfulness of RT-PCR assays for PVY and PLRV.
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Using the Favret, PVY and PLRV primer sets, single aphids of M. persicase, M. euphorbiae and A. nasturtii that had been exposed to PLRV-infected and/or PVY-infected potato plants were assessed for their acquisition of the viruses by the triplex RT-PCR assay.
Importantly, because aphids reproduce parthenogenetically, we were able to use a single aphid genotype, such that confounding effects of genetic variation among comparisons between stress types did not influence results.
The sensitivity of the RLMV assay was also tested on single aphids after a fixed acquisition access period (AAP).
To maintain the same density of aphids we added six of the same species for the single aphid treatments, or three of each species for the paired aphid treatments (substitutive design).
Microsatellite markers developed by Kim et al. (2010) were used to confirm this clonal colony consisted of a single aphid genotype.
Some of the samples consisted of a single aphid collected directly from the source tree or plant and stored as above.
One aphid line consisted of the genetically identical progeny of a single aphid.
Mounds often only harbored one clonal lineage of a single aphid species and if mounds had multiple aphid clones they were almost always compartmentalized in different chambers.
Values of the probability of transmission of CMV isolates in melon by a single aphid.
DP, double-parasitism; Hd, H. defensa We dissected parasitized aphids to verify that the number of observed parasitism events (single vs. double) corresponds to the number of eggs deposited in a single aphid host.
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