Biogenic VOCs Emission Profiles Associated with Plant-Pest Interaction for Phenotyping Applications.

Pest attacks on plants can substantially change plants' volatile organic compounds (VOCs) emission profiles. Comparison of VOC emission profiles between non-infected/non-infested and infected/infested plants, as well as resistant and susceptible plant cultivars, may provide cues for a deeper understanding of plant-pest interactions and associated resistance. Furthermore, the identification of biomarkers-specific biogenic VOCs-associated with the resistance can serve as a non-destructive and rapid tool for phenotyping applications. This research aims to compare the VOCs emission profiles under diverse conditions to identify constitutive (also referred to as green VOCs) and induced (resulting from biotic/abiotic stress) VOCs released in potatoes and wheat. In the first study, wild potato Solanum bulbocastanum (accession# 22; SB22) was inoculated with Meloidogyne chitwoodi race 1 (Mc1), and Mc1 pathotype Roza (SB22 is resistant to Mc1 and susceptible to pathotype Roza), and VOCs emission profiles were collected using gas chromatography-flame ionization detection (GC-FID) at different time points. Similarly, in the second study, the VOCs emission profiles of resistant ('Hollis') and susceptible ('Alturas') wheat cultivars infested with Hessian fly insects were evaluated using the GC-FID system. In both studies, in addition to variable plant responses (susceptibility to pests), control treatments (non-inoculated or non-infested) were used to compare the VOCs emission profiles resulting from differences in stress conditions. The common VOC peaks (constitutive VOCs) between control and infected/infested samples, and unique VOC peaks (induced VOCs) presented only in infected/infested samples were analyzed. In the potato-nematode study, the highest unique peak was found two days after inoculation (DAI) for SB22 inoculated with Mc1 (resistance response). The most common VOC peaks in SB22 inoculated with both Mc1 and Roza were found at 5 and 10 DAI. In the wheat-insect study, only the Hollis showed unique VOC peaks. Interestingly, both cultivars released the same common VOCs between control and infected samples, with only a difference in VOC average peak intensity at 22.4 min retention time where the average intensity was 4.3 times higher in the infested samples of Hollis than infested samples of Alturas. These studies demonstrate the potential of plant VOCs to serve as a rapid phenotyping tool to assess resistance levels in different crops.

Interspecific interactions within a vector-borne complex are influenced by a co-occurring pathosystem.

Potato virus Y (PVY) and zebra chip (ZC) disease are major threats to solanaceous crop production in North America. PVY can be spread by aphid vectors and through vegetative propagation in potatoes. ZC is associated with "Candidatus Liberibacter solanacearum" (Lso), which is transmitted by the tomato/potato psyllid, Bactericera cockerelli Sulc (Hemiptera: Triozidae). As these two pathosystems may co-occur, we studied whether the presence of one virus strain, PVY°, affected the host preference, oviposition, and egg hatch rate of Lso-free or Lso-carrying psyllids in tomato plants. We also examined whether PVY infection influenced Lso transmission success by psyllids, Lso titer and plant chemistry (amino acids, sugars, and phytohormones). Lso-carrying psyllids showed a preference toward healthy hosts, whereas the Lso-free psyllids preferentially settled on the PVY-infected tomatoes. Oviposition of the Lso-carrying psyllids was lower on PVY-infected than healthy tomatoes, but Lso transmission, titer, and psyllid egg hatch were not significantly affected by PVY. The induction of salicylic acid and its related responses, and not nutritional losses, may explain the reduced attractiveness of the PVY-infected host to the Lso-carrying psyllids. Although our study demonstrated that pre-existing PVY infection can reduce oviposition by the Lso-carrying vector, the preference of the Lso-carrying psyllids to settle on healthy hosts could contribute to Lso spread to healthy plants in the presence of PVY infection in a field.

Effect of the level of "Candidatus Liberibacter solanacearum" infection on the development of zebra chip disease in different potato genotypes at harvest and post storage.

Potato psyllid (Bactericera cockerelli Sulc)-transmitted "Candidatus Liberibacter solanacearum" (Lso) has been negatively impacting the potato industry in the United States as well as other potato-producing countries. Lso has been linked to a condition known as zebra chip (ZC) that affects yield and quality of potato tubers. Efforts to find sources of resistance to ZC have primarily focused on greenhouse evaluations based on a single inoculation time prior to harvest. Plant response to infection, however, could be influenced by the developmental stage of the host plant, and ZC may continue to develop after harvest. The objectives of this study were to quantify Lso inoculation success, Lso titer, ZC severity and Lso development during storage in eight potato genotypes. These evaluations were conducted on plants infested with Lso-positive psyllids at 77, 12, and 4 days before vine removal (DBVR). The evaluated genotypes were categorized according to their relative resistance to Lso and tolerance to ZC symptoms. Lso inoculation success in the genotype family A07781, derived from Solanum chacoense, was lower than that of the susceptible control ('Russet Burbank'). A07781-4LB and A07781-3LB genotypes were characterized relatively resistant to the pathogen and highly tolerant to ZC symptoms, while A07781-10LB was categorized as susceptible to Lso but relatively tolerant to symptom expression. In stored potatoes, increase in Lso concentrations was observed for all infestation times. However, significantly higher Lso titer was detected in tubers infested 12 DBVR and the effect was similar across genotypes. Overall, the A07781 family can be considered as a promising source of resistance or tolerance to ZC.

Potato virus Y Transmission Efficiency from Potato Infected with Single or Multiple Virus Strains.

There has been a recent shift in the prevalence of Potato virus Y (PVY) strains affecting potato with the ordinary strain PVYO declining and the recombinant strains PVYNTN and PVYN:O emerging in the United States. Multiple PVY strains are commonly found in potato fields and even in individual plants. Factors contributing to the emergence of the recombinant strains are not well defined but differential aphid transmission of strains from single and mixed infections may play a role. We found that the transmission efficiencies by Myzus persicae, the green peach aphid, of PVYNTN, PVYN:O, and PVYO varied depending on the potato cultivar serving as the virus source. Overall transmission efficiency was highest from sources infected with three virus strains, whereas transmission from sources infected with one or two virus strains was not significantly different. Two strains were concomitantly transmitted by individual aphids from many of the mixed-source combinations, especially if PVYO was present. Triple-strain infections were not transmitted by any single aphid. PVYO was transmitted most efficiently from mixed-strain infection sources. The data do not support the hypothesis that differential transmission of PVY strains by M. persicae is a major contributing factor in the emergence of recombinant PVY strains in the U.S. potato crop.

Responses of Aphid Vectors of Potato leaf roll virus to Potato Varieties.

Potato leaf roll virus (PLRV) can reduce tuber yield and quality in potato. Green peach aphid (Myzus persicae [Sulzer]) and potato aphid (Macrosiphum euphorbiae [Thomas]) are the two most important potato-colonizing PLRV vectors in the Pacific Northwest. We compared My. persicae and Ma. euphorbiae densities and PLRV incidences among potato varieties in the field to clarify the relationships between aphid abundance and PLRV incidence in plants. Aphids were sampled weekly over three years in the potato varieties Russet Burbank, Ranger Russet, and Russet Norkotah in a replicated field trial. In all years, My. persicae was more abundant than Ma. euphorbiae, representing at least 97% of samples. My. persicae densities did not differ among potato varieties across years; very low numbers of Ma. euphorbiae precluded such statistical comparisons for this species. PLRV infection did not differ significantly among potato varieties, although the percent of PLRV-infected plants differed among years when all varieties were combined (46% in 2013, 29% in 2011, 13% in 2012). For Ranger Russet and Russet Norkotah, PLRV incidence was positively correlated with aphid abundance as well as proportion of PLRV-positive aphids. In Russet Burbank, only aphid abundance was positively correlated with PLRV infection. Our results suggest that the three most commonly grown potato varieties in our region do not differ in their susceptibility to PLRV infection, and that aphid density was a consistent indicator of the risk of infection by this virus across varieties. Both of these findings can be used to hone PLRV monitoring and modeling efforts.

Contribution of Noncolonizing Aphids to Potato Virus Y Prevalence in Potato in Idaho.

Potato virus Y (PVY) is a major concern for potato production in the United States given its impact on both crop quality and yield. Although green peach aphid, Myzus persicae (Sulzer), is the most efficient PVY vector, it may be less abundant in potato-growing areas of Idaho relative to non-potato-colonizing aphid vectors of PVY that may disperse from nearby cereal fields and other crops. A field study was conducted during 2012-2013 to examine if noncolonizing aphids disperse to nearby potato fields as cereal crops dry down before harvest. The aphid fauna was sampled weekly in four different potato fields in south-central and southeastern Idaho using yellow sticky traps and yellow pan traps. Potato fields were chosen with an adjacent cereal field such that the prevailing westerly wind would facilitate aphid dispersal from cereal fields to potato. Non-potato-colonizing aphids sampled included 10 cereal aphid species, the most abundant of which were Rhopalosiphum padi L. and Metopolophium dirhodum (Walker). More than 35 species from noncereal hosts also were found. Overall, green peach aphid abundance was relatively low, ranging from 0.5-2.5% of the total aphid capture between years and among fields. In both years and all locations, cereal aphid abundance peaked in mid- to late July (cereal ripening stage) and decreased thereafter as cereal crops dried. PVY prevalence in the potato fields increased following these increases in aphid abundance. This study suggests that cereal aphids and other noncolonizing aphids are important contributors to PVY prevalence in potato in southern Idaho.

Interactions among potato genotypes, growth stages, virus strains, and inoculation methods in the potato virus Y and green peach aphid pathosystem.

Potato virus Y (PVY) is an economically important and reemerging potato pathogen in North America. PVY infection reduces yield, and some necrotic and recombinant strains render tubers unmarketable. Although PVY(O) is the most prevalent strain in the United States, the necrotic and recombinant strains PVY(NTN) and PVY(N:O) are becoming more widespread. Infection rates in aphid-inoculated (Myzus persicae (Sulzer)) and mechanically inoculated plants were compared across two potato genotypes ('Yukon Gold' and A98345-1), three PVY strains (PVY(O), PVY(N:O), and PVY(NTN)), and two growth stages at inoculation (pre- and postflowering). Susceptibility of genotypes was measured as infection rate using a double-antibody sandwich-enzyme-linked immunosorbent assay; virus titer and tuber mass also were recorded from the infected plants. Yukon Gold generally was more susceptible than A98345-1 to all three PVY strains, especially following mechanical inoculation. Within genotypes, Yukon Gold was most susceptible to PVY(O) and A98345-1 was most susceptible to PVY(N:O). Plants exhibited age-based resistance, with both genotypes showing higher susceptibility at the pre- than postflowering stage. The overall ranking pattern of virus titer in infected plants was PVY(O) > PVY(NTN) > PVY(N:O); across all three strains, infected Yukon Gold had higher titer than infected A98345-1 plants. Yukon Gold plants had lower tuber mass than A98345-1 when infected, and there were differences between the two inoculation methods in regard to tuber mass for the three stains. The results showed differences in infection response between inoculation methods and as a function of genotype, strain, inoculation stage, and their interactions. These factors should be considered when screening genotypes for resistance.

Preference by a virus vector for infected plants is reversed after virus acquisition.

Pathogens and their vectors can interact either directly or indirectly via their shared hosts, with implications for the persistence and spread of the pathogen in host populations. For example, some plant viruses induce changes in host plants that cause the aphids that carry these viruses to settle preferentially on infected plants. Furthermore, relative preference by the vector for infected plants can change to a preference for noninfected plants after virus acquisition by the vector, as has recently been demonstrated in the wheat-Rhopalosiphum padi-Barley yellow dwarf virus pathosystem. Here we document a similar dynamic in the potato-Myzus persicae (Sulzer)-Potato leaf roll virus (PLRV) pathosystem. Specifically, in a dual choice bioassay, nonviruliferous apterous M. persicae settled preferentially on or near potato plants infected with PLRV relative to noninfected (sham-inoculated) control plants, whereas viruliferous M. persicae (carrying PLRV) preferentially settled on or near sham-inoculated potato plants relative to infected plants. The change in preference after virus acquisition also occurred in response to trapped headspace volatiles, and to synthetic mimics of headspace volatile blends from PLRV-infected and sham-inoculated potato plants. The change in preference we document should promote virus spread by increasing rates of virus acquisition and transmission by the vector.

The influence of virus-induced changes in plants on aphid vectors: insights from luteovirus pathosystems.

Plant virus infection can alter the suitability of host plants for their aphid vectors. Most reports indicate that virus-infected plants are superior hosts for vectors compared to virus-free plants with respect to vector growth rates, fecundity and longevity. Some aphid vectors respond preferentially to virus-infected plants compared to virus-free ones, while others avoid infected plants that are inferior hosts. Thus, it appears vectors can exploit changes in host plant quality associated with viral infection. Enhanced vector performance and preference for virus-infected plants might also be advantageous for viruses by promoting their spread and possibly enhancing their fitness. Our research has focused on two of the most important luteoviruses that infect wheat (Barley yellow dwarf virus), or potato (Potato leafroll virus), and their respective aphid vectors, the bird-cherry oat aphid, Rhopalosiphum padi, and the green peach aphid, Myzus persicae. The work has demonstrated that virus infection of host plants enhances the life history of vectors. Additionally, it has shown that virus infection alters the concentration and relative composition of volatile organic compounds in host plants, that apterae of each vector species settle preferentially on virus-infected plants, and that such responses are mediated by volatile organic compounds. The findings also indicate that plants respond heterogeneously to viral infection and as a result different plant parts change in attractiveness to vectors during infection and vector responses to virus-infected plants are dynamic. Such dynamic responses could enhance or reduce the probability of virus acquisition by individual aphids searching among plants. Finally, our work indicates that compared to non-viruliferous aphids, viruliferous ones are less or not responsive to virus-induced host plant volatiles. Changes in vector responsiveness to plants after vectors acquire virus could impact virus epidemiology by influencing virus spread. The potential implications of these findings for virus ecology and epidemiology are discussed.

Changes in green peach aphid responses to potato leafroll virus-induced volatiles emitted during disease progression.

Previous research has shown that green peach aphids, Myzus persicae (Sulzer), preferentially settle on leaflets of potato plants (Solanum tuberosum L.) infected with potato leafroll virus (PLRV) compared with sham-inoculated controls, at least in part because of aphid responses to volatile cues from the plants. The prior work used plants 4 wk after inoculation. In this study, aphid emigration from the vicinity of leaflets of PLRV-infected plants at 2, 4, 6, 8, and 10 wk after inoculation was compared with emigration from leaflets of sham-inoculated control plants. For the bioassay, 30 aphids were placed directly above a test leaflet on screening to exclude gustatory and tactile cues and in darkness to exclude visual cues. The numbers emigrating were recorded every 10 min for 1 h. Volatile organic compounds (VOCs) were collected from the headspace of the test plants, quantified, and compared among treatments. In bioassays with leaflets of upper nodes of the plants, aphid immigration rates were significantly lower from leaflets of PLRV-infected plants than from sham-inoculated plants at 4 and 6 wk after inoculation, but not at 2, 8, and 10 wk after inoculation. In bioassays with leaflets from lower nodes, emigration did not differ between PLRV-infected plants and sham-inoculated plants at any stage in the infection. Volatile compounds detectable in the headspace of intact plants at 2, 4, and 8 wk after inoculation (or sham inoculation) changed with plant age and with disease progression, potentially explaining behavioral responses of the aphids.

Effect of an alternate weed host, hairy nightshade, Solanum sarrachoides, on the biology of the two most important potato leafroll virus (Luteoviridae: Polerovirus) vectors, Myzus persicae and Macrosiphum euphorbiae (Aphididae: Homoptera).

Hairy nightshade, Solanum sarrachoides (Sendtner), is a ubiquitous weed in potato agro-ecosystems and nonagricultural lands of southeastern Idaho and the Pacific Northwest. This weed increases the complexity of the Potato leafroll virus (PLRV) (Luteoviridae: Polervirus)-potato pathosystem by serving as aphid and virus reservoir. Previous field studies showed higher densities of green peach aphid, Myzus persicae (Sulzer), and potato aphid, Macrosiphum euphorbiae (Thomas), the two most important vectors of PLRV, on S. sarrachoides compared with potato plants in the same fields. Some of the S. sarrachoides plants sampled in these surveys tested positive for PLRV. Viral infections can alter the physiology of plant hosts and aphid performance on such plants. To understand better the potential effects of S. sarrachoides on the PLRV-potato-aphid pathosystem, the life histories of M. persicae and M. euphorbiae were compared on virus-free and PLRV-infected S. sarrachoides and potato. Individual nymphs of each aphid species were held in clip cages on plants from each treatment to monitor their development, survival, and reproductive output. Nymphal survival for both aphids across plant species was higher on S. sarrachoides than on potato, and, within plant species, it was higher on PLRV-infected plants than on noninfected plants. With a few exceptions, similar patterns occurred for fecundity, reproductive periods, adult longevity, and intrinsic rate of increase. The enhanced performance of aphids on S. sarrachoides and on PLRV-infected plants could alter the vector population dynamics and thus the PLRV-disease epidemiology in fields infested with this weed.

Myzus persicae is arrested more by blends than by individual compounds elevated in headspace of PLRV-infected potato.

Volatiles from potato plants (Solanum tuberosum L.) infected with Potato leaf roll virus (PLRV) attract and arrest the principal vector of PLRV, the green peach aphid, Myzus persicae (Sulzer), more strongly than volatiles from non-infected plants. The total concentration of volatiles detectable in the headspace of PLRV-infected plants is greater than that in the headspace of non-infected controls, and the relative composition is altered. To determine the basis of the aphid response to PLRV-infection-induced volatiles from potato, behavioral bioassays were conducted. We measured arrestment of aphids by individual components, by synthetic blends of these, and by a naturally occurring blend by using an emigration rate bioassay, and quantified observations of the behavior of individual aphids. The components tested were those elevated at least twofold in response to PLRV infection. Before conducting the behavioral bioassays, electroantennograms confirmed the electrophysiological responses of aphids to the components of the blend. For bioassays, individual compounds or blends were tested by applying them in solution to paper strips at concentrations designed to mimic those present in the headspace of the plants. All bioassays were conducted by placing aphids on fine-mesh screening positioned above treated paper strips. Arrestment was measured by placing groups of 30 aphids directly over the treated strips and counting the number moving away at 10-min intervals for 50 min. Among the individual compounds tested, only beta-pinene was a mild arrestant. The other compounds did not elicit significant changes in arrestment or behavior at a range of physiologically relevant concentrations. In contrast, synthetic blends that mimicked the concentration and composition present in headspace of PLRV-infected potato plants arrested aphids significantly more strongly than blends mimicking volatiles from the headspace of non-infected plants. The naturally occurring blend collected from headspace of PLRV-infected plants also arrested M. persicae more strongly than the blend collected from headspace of non-infected plants. Aphid behavior was quantified by directly observing individual aphids and recording their activities during a 5-min period on screening above strips treated with test materials. Few differences in time budgets were observed among aphids exposed to individual components, but synthetic blends and trapped headspace volatiles from PLRV-infected plants resulted in significantly less time spent walking by aphids than synthetic blends and trapped headspace from non-infected controls. Our results indicate that arrestment of M. persicae by PLRV-infected plants requires the blend of volatile organic compounds released by these plants and is not produced in response to a single compound.