Assessing the Temporal Effects of Squash vein yellowing virus Infection on Settling and Feeding Behavior of Bemisia tabaci (MEAM1) (Hemiptera: Aleyrodidae).

Insect vector behavior and biology can be affected by pathogen-induced changes in the physiology and morphology of the host plant. Herein, we examined the temporal effects of Squash vein yellowing virus (family Potyviridae, genus Ipomovirus) infection on the settling, oviposition preference, and feeding behavior of its whitefly vector, Bemisia tabaci (Gennadius) Middle East-Asia Minor 1 (MEAM1), formerly known as B. tabaci biotype B. Settling and oviposition behavioral choice assays were conducted on pairs of infected and mock-inoculated watermelon (Citrullus lanatus (Thunb) Matsum and Nakai) (Cucurbitales: Cucurbitaceae) at 5-6 days post inoculation (DPI) and 10-12 DPI. Electropenetrography, or electrical penetration graph (both abbreviated EPG), was used to assess differences in feeding behaviors of whitefly on mock-inoculated, 5-6 and 10-12 DPI infected watermelon plants. Whiteflies showed no preference in settling or oviposition on the infected and mock-inoculated plants at 5-6 DPI. However, at 10-12 DPI, whiteflies initially settled on infected plants but then preference of settling shifted to mock-inoculated plants after 8 h. Only at 10-12 DPI, females laid significantly more eggs on mock-inoculated plants than infected plants. EPG revealed no differences in whitefly feeding behaviors among mock-inoculated, 5-6 DPI infected and 10-12 DPI infected plants. The results highlighted the need to examine plant disease progression and its effect on vector behavior and performance, which could play a crucial role in Squash vein yellowing virus spread.

Accumulation of terpenoid phytoalexins in maize roots is associated with drought tolerance.

Maize (Zea mays) production, which is of global agro-economic importance, is largely limited by herbivore pests, pathogens and environmental conditions, such as drought. Zealexins and kauralexins belong to two recently identified families of acidic terpenoid phytoalexins in maize that mediate defence against both pathogen and insect attacks in aboveground tissues. However, little is known about their function in belowground organs and their potential to counter abiotic stress. In this study, we show that zealexins and kauralexins accumulate in roots in response to both biotic and abiotic stress including, Diabrotica balteata herbivory, Fusarium verticillioides infection, drought and high salinity. We find that the quantity of drought-induced phytoalexins is positively correlated with the root-to-shoot ratio of different maize varieties, and further demonstrate that mutant an2 plants deficient in kauralexin production are more sensitive to drought. The induction of phytoalexins in response to drought is root specific and does not influence phytoalexin levels aboveground; however, the accumulation of phytoalexins in one tissue may influence the induction capacity of other tissues.

Differential Probing Behavior of Blissus insularis (Hemiptera: Blissidae) on Resistant and Susceptible St. Augustinegrasses.

Southern chinch bug, Blissus insularis Barber, is a severe pest of St. Augustinegrass throughout the southern United States. Host plant resistance is an environmentally friendly method to manage chinch bug infestations and is increasingly important, as the southern chinch bug develops resistance to insecticides. In this study, in an effort to understand resistance mechanisms in two varieties of St. Augustinegrass ('FX-10' and 'NUF-76'), we used the electrical penetration graph method to quantify stylet probing behaviors in two resistant and two susceptible St. Augustinegrass varieties. Overall, chinch bugs spent less time probing on resistant FX-10 and NUF-76 than on susceptible 'Floratam' and 'Palmetto', and individual probes were shorter in average duration but more numerous in resistant varieties than in susceptible varieties. During probing, chinch bugs spent more time in pathway-associated stylet activities (i.e., penetration through epidermal and mesophyll tissue) in the resistant varieties than in the susceptible varieties, likely indicating difficulty in finding and accessing an ingestion site. As a consequence, chinch bugs spent proportionately much less time engaged in xylem ingestion in both resistant varieties than in susceptible varieties but only in FX-10 were phloem-associated activities significantly reduced compared with those in susceptible varieties. We conclude that there is evidence for non-phloem-associated chinch-bug resistance factors in both NUF-76 and FX-10, and phloem-associated factors in FX-10.

Oxidative responses of St. Augustinegrasses to feeding of southern chinch bug, Blissus insularis Barber.

Southern chinch bug, Blissus insularis Barber (Hemiptera: Blissidae), is a serious insect pest of St. Augustinegrass, Stenotaphrum secundatum (Walt.) Kuntze, a turfgrass commonly grown in the southeastern United States. Resistance to southern chinch bug has been identified in the polyploid St. Augustinegrass varieties 'Floratam' and 'FX-10', and the diploid 'Captiva'. However, southern chinch bug in Florida and elsewhere has overcome Floratam's resistance. This research investigated the potential role of selected plant oxidative enzymes in resistance/susceptibility to southern chinch bug in two polyploid varieties (FX-10 and Floratam) and two diploid varieties (Captiva and Palmetto). Oxidative enzyme activity was estimated spectrophotometrically from plant samples collected 1, 3, 5, and 8 days after southern chinch bug infestation and from uninfested control plants. Resistant FX-10 and Captiva had significantly higher peroxidase activity, while Captiva had significantly higher polyphenol oxidase activity 5 and 8 days after infestation compared to uninfested controls. FX-10 had higher lipoxygenase activity 3, 5, and 8 days after infestation compared to uninfested controls. Catalase activities did not differ between infested and control plants in any of the varieties tested. Native gels stained for peroxidase indicated that certain isozymes in FX-10 and Captiva were induced 5 and 8 days after infestation. Isozyme profiles of polyphenol oxidase and lipoxygenase did not differ between control and infested FX-10, Floratam, Captiva, and Palmetto. Potential mechanisms to explain the correlation of resistance to southern chinch bug in FX-10 and Captiva with higher activities of oxidative enzymes are discussed.

Enzyme induction as a possible mechanism for latex-mediated insect resistance in romaine lettuce.

Plant latex is a known storehouse of various secondary metabolites with demonstrated negative impact on insect fitness. A romaine lettuce cultivar, "Valmaine", possesses a high level of latex-mediated resistance against the banded cucumber beetle, Diabrotica balteata LeConte (Coleoptera: Chrysomelidae), compared to a closely related cultivar "Tall Guzmaine". Latex from damaged Valmaine plants was much more deterrent to adult D. balteata feeding than latex from undamaged plants when applied to the surface of artificial diet under choice conditions; no such difference was found in choice tests with latex from damaged and undamaged Tall Guzmaine plants. The intensities of whiteness and browning were significantly higher in Valmaine latex than in Tall Guzmaine latex. The activities of three enzymes (phenylalanine ammonia lyase, polyphenol oxidase, and peroxidase) significantly increased over time in latex from damaged Valmaine plants (i.e., 1, 3, and 6 days after feeding initiation), but they remained the same in Tall Guzmaine latex. The constitutive levels of phenylalanine ammonia lyase and polyphenol oxidase also were significantly higher in Valmaine latex than in Tall Guzmaine latex. These studies suggest that Valmaine latex chemistry may change after plant damage due to increased activity of inducible enzymes and that inducible resistance appears to act synergistically with constitutive resistance against D. balteata.

Pepper weevil attraction to volatiles from host and nonhost plants.

The location of wild and cultivated host plants by pepper weevil (Anthonomus eugenii Cano) may be aided by visual cues, the male-produced aggregation pheromone, herbivore-induced, or constitutive host plant volatiles. The attractiveness of constitutive plant volatiles to pioneer weevils is important in understanding, and perhaps controlling, dispersal of this insect between wild and cultivated hosts. Ten-day-old male and 2- and 10-day-old female weevils were tested in short-range Y-tube assays. Ten-day-old male and female weevils were attracted to the volatiles released by whole plants of three known oviposition hosts, 'Jalapeno' pepper, American black nightshade, and eggplant, as well as tomato, a congener, which supports feeding but not oviposition. Two-day-old females were attracted to all plants tested, including lima bean, an unrelated, nonhost plant. Fruit volatiles from all three hosts and flower volatiles from nightshade and eggplant were also attractive. In choice tests, weevils showed different preferences for the oviposition hosts, depending on age and sex. Upwind response of 10-day-old male and female weevils to host plant volatiles was also tested in long-range wind tunnel assays. Weevils responded to pepper, nightshade, and eggplant volatiles by moving upwind. There was no difference in the observed upwind response of the weevils to the three host plants under no-choice conditions. Reproductively mature pepper weevils can detect, orient to, and discriminate between the volatile plumes of host plants in the absence of visual cues, conspecific feeding damage, or the presence of their aggregation pheromone.

Role of leaf sheath lignification and anatomy in resistance against southern chinch bug (Hemiptera: Blissidae) in St. Augustinegrass.

Southern chinch bug, Blissus insularis Barber (Hemiptera: Blissidae), is the most serious insect pest of St. Augustinegrass Stenotaphrum secundatum (Walter) Kuntze, a common lawngrass grown in southeastern U.S. states. Host plant resistance to southern chinch bug has been identified in the polyploid St. Augustinegrass 'FX-10' and the diploid 'Captiva'. The objective of this research was to identify possible physical mechanism(s) explaining chinch bug resistance in these cultivars. We studied the distribution of chinch bug salivary sheaths in the preferred tissue for feeding (the axillary shoot) of the two resistant cultivars and two susceptible cultivars, paired for ploidy ('Floratam', polyploid, and Palmetto, diploid). We also investigated the potential role of axillary shoot lignification and anatomy in chinch bug resistance. Salivary sheaths were more abundant on the outermost leaf sheath of axillary shoots of resistant cultivars compared with susceptible cultivars. In contrast, fewer salivary sheaths reached the innermost meristematic tissue in the axillary shoots of resistant St. Augustinegrass cultivars than in the two susceptible cultivars. The polyploid cultivars FX-10 and Floratam had higher total lignin in axillary shoots compared with the diploid cultivars Captiva and Palmetto. However, total lignin content was not correlated with resistance to southern chinch bug. Light microscopic studies found no differences in epidermal layer thickness among resistant and susceptible St. Augustinegrass cultivars. However, transmission electron microscopic studies revealed that the cell walls of the sclerenchyma cells around the vascular bundle of southern chinch bug-resistant FX-10 and Captiva were significantly thicker than the cell walls in susceptible Floratam and Palmetto. Our research suggests that the thick-walled sclerenchyma cells around the vascular bundle play a role in southern chinch bug resistance in St. Augustinegrass, possibly by reducing stylet penetration to the vascular tissue.

Host-Mediated Effects of Semipersistently Transmitted Squash Vein Yellowing Virus on Sweetpotato Whitefly (Hemiptera: Aleyrodidae) Behavior and Fitness.

Plant viruses may indirectly affect insect vector behavior and fitness via a shared host plant. Here, we evaluated the host-mediated effects of Squash vein yellowing virus (SqVYV) on the behavior and fitness of its whitefly vector, Bemisia tabaci (Gennadius) Middle East-Asia Minor 1, formerly biotype B. Alighting, settling, and oviposition behavioral assays were conducted on infected and mock-inoculated squash (Cucurbita pepo L.) and watermelon [Citrullus lanatus (Thunb) Matsum and Nakai] plants. Developmental time of immature stages, adult longevity, and fecundity were measured on infected and mock-inoculated squash plants. For adult longevity and fecundity, whiteflies were reared on infected and mock-inoculated squash plants to determine the effects of nymphal rearing host on the adult stage. More whiteflies alighted and remained settled on infected squash than on mock-inoculated squash 0.25, 1, 8, and 24 h after release. No such initial preference was observed on watermelon plants, but by 8 h after release, more whiteflies were found on mock-inoculated watermelon plants than on infected plants. Whiteflies laid approximately six times more eggs on mock-inoculated watermelon than on infected watermelon; however, no differences were observed on squash. Development from egg to adult emergence was 3 d shorter on infected than mock-inoculated squash plants. Females lived 25% longer and had higher fecundity on infected squash plants than on mock-inoculated plants, regardless of infection status of the rearing host. The host-mediated effects of SqVYV infection on whitefly behavior differ on two cucurbit host plants, suggesting the potential for more rapid spread of the virus within watermelon fields.

Host plant resistance in romaine lettuce affects feeding behavior and biology of Trichoplusia ni and Spodoptera exigua (Lepidoptera: Noctuidae).

Lettuce quality and yield can be reduced by feeding of several lepidopterous pests, particularly cabbage looper, Trichoplusia ni (Hübner), and beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae). Host plant resistance to these insects is an environmentally sound adjunct to conventional chemical control. In this study we compared the survival, development, and feeding behavior of cabbage looper and beet armyworm on two romaine lettuce cultivars, resistant 'Valmaine' and susceptible 'Tall Guzmaine'. Larval mortality of both species was significantly higher on resistant Valmaine than on susceptible Tall Guzmaine. The average weight per larva after feeding for 1 wk on Tall Guzmaine plants was 6 times (beet armyworm) and 2 times (cabbage looper) greater than that of larvae feeding on Valmaine plants. Significant reduction in larval growth on Valmaine compared with that on Tall Guzmaine resulted in a 5.9- (beet armyworm) and 2.6-d (cabbage looper) increase in larval duration and almost a 1-d increase in pupal duration. Average pupal and adult weights and successful pupation of cabbage looper and beet armyworm were reduced on Valmaine compared with Tall Guzmaine. The sex ratio of progeny did not deviate from 1:1 when larvae were reared on either Valmaine or Tall Guzmaine. The fecundity of cabbage looper and beet armyworm adults that developed from larvae reared on Valmaine was about one-third that of adults from Tall Guzmaine, but adult longevity did not significantly differ on the two lettuce cultivars. The two insect species showed different feeding preferences for leaves of different age groups on Valmaine and Tall Guzmaine. Cabbage loopers cut narrow trenches on the leaf before actual feeding to block the flow of latex to the intended site of feeding. In contrast, beet armyworms did not trench. The different feeding behavior of the two species on Valmaine may explain the superior performance of cabbage looper compared with beet armyworm.

Interactive Effects of Elevated [CO2] and Drought on the Maize Phytochemical Defense Response against Mycotoxigenic Fusarium verticillioides.

Changes in climate due to rising atmospheric carbon dioxide concentration ([CO2]) are predicted to intensify episodes of drought, but our understanding of how these combined conditions will influence crop-pathogen interactions is limited. We recently demonstrated that elevated [CO2] alone enhances maize susceptibility to the mycotoxigenic pathogen, Fusarium verticillioides (Fv) but fumonisin levels remain unaffected. In this study we show that maize simultaneously exposed to elevated [CO2] and drought are even more susceptible to Fv proliferation and also prone to higher levels of fumonisin contamination. Despite the increase in fumonisin levels, the amount of fumonisin produced in relation to pathogen biomass remained lower than corresponding plants grown at ambient [CO2]. Therefore, the increase in fumonisin contamination was likely due to even greater pathogen biomass rather than an increase in host-derived stimulants. Drought did not negate the compromising effects of elevated [CO2] on the accumulation of maize phytohormones and metabolites. However, since elevated [CO2] does not influence the drought-induced accumulation of abscisic acid (ABA) or root terpenoid phytoalexins, the effects elevated [CO2] are negated belowground, but the stifled defense response aboveground may be a consequence of resource redirection to the roots.

Repellency of ginger oil to Bemisia argentifolii (Homoptera: Aleyrodidae) on tomato.

The whitefly Bemisia argentifolii Bellows & Perring is an economically important pest of tomatoes, Lycopersicon esculentum Mill., inducing an irregular ripening disorder of fruit and transmitting plant pathogenic viruses. With the goal of investigating ginger oil as a protectant for tomato plants, we tested the effects of concentration of ginger oil and application methods on repellency to whitefly in a vertical still-air olfactometer. In choice and no-choice experiments conducted in a greenhouse, we evaluated whether ginger oil would protect tomato seedlings from whitefly settling and oviposition. Ginger oil repelled whitefly adults in the vertical olfactometer. The repellency of ginger oil was attributed to its odor, effective at the concentrations used over a distance of 1-2 mm. Tomato leaf disks dipped in ginger oil repelled whiteflies at concentrations of 0.5, 0.75, and 1%, but not at concentrations <0.5%, in a dose-response experiment conducted in the olfactometer. Repellency increased with increasing ginger oil concentration when leaf disks were dipped in ginger oil but not when ginger oil was sprayed onto the leaf disks. Higher quantities of monoterpenes and sesquiterpenes were deposited on leaf disks dipped in ginger oil than on sprayed leaf disks according to gas chromatographic quantification. In the greenhouse, both choice and no-choice tests were conducted with tomato seedlings dipped in 0.25% ginger oil solution or 2% Tween 20, as treatment and control, respectively. In the choice test, 35-42% fewer whitefly adults settled and 37% fewer eggs were laid during the 24-h exposure period on tomato plants dipped in ginger oil solution than on plants dipped in 2% Tween 20. In the no-choice test, 10.2-16.7% fewer whiteflies settled on treated plants compared with control plants but no significant differences were detected in the number of eggs laid. Higher concentrations of ginger oil could not be used without causing severe wilting of tomato leaves. Ginger oil has potential as a protectant of tomato seedlings against B. argentifolii, but issues of phytotoxicity and coverage need to be addressed.

Influence of Bemisia argentifolii (Homoptera: Aleyrodidae) infestation and squash silverleaf disorder on zucchini seedling growth.

We investigated the effect of different levels of infestation by whiteflies, Bemisia argentifolii Bellows & Perring, on the growth and pigment concentrations of seedlings of zucchini, Cucurbita pepo L., that differed in their tolerance to squash silverleaf disorder. Genetically similar sister lines that were either tolerant (ZUC76-SLR) or susceptible (ZUC61) to silverleaf disorder exhibited reduced plant height, internode length, plant dry weight, and petiole length in response to whitefly feeding. Similar plant growth responses to whitefly feeding were observed despite that the foliage of ZUC61 silvered severely, whereas the foliage of ZUC76-SLR showed no silvering in a greenhouse experiment conducted in the spring and showed only minimal silvering in a similar greenhouse experiment conducted in the fall. In plants of both sister lines infested with 50 pairs of whiteflies and their progeny, petioles, but not the leaf blades, of uninfested leaves had reduced chlorophyll content. In another experiment, two different genetic sources of tolerance to silverleaf disorder (ZUC33-SLR/PMR and ZUC76-SLR) and a commercial silverleaf-susceptible zucchini hybrid ('Zucchini Elite') responded similarly to whitefly feeding, except the tolerant genotypes did not exhibit leaf silvering. All genotypes, silverleaf tolerant or not, had reduced dry weight, plant height, and internode length that became more pronounced as whitefly infestation increased. All genotypes had reduced levels of chlorophylls and carotenoids in uninfested young leaf blades and petioles from infested plants. Petioles, however, were more affected by feeding than leaf blades, showing a 66% reduction in chlorophylls a+b and carotenoids at the lowest infestation level (30 pairs of whitefly and their progeny), whereas pigments in leaf blades declined more slowly in response to whitefly feeding density, averaging 14-15% less at the highest infestation level (90 pairs of whitefly and their progeny). We conclude that tolerance to silverleaf disorder does not prevent stunting in zucchini seedlings nor does it protect against the systemic loss of photosynthetic and protoprotectant pigments induced by feeding of B. argentifolii whiteflies.

Resistance to adult banded cucumber beetle (Coleoptera: Chrysomelidae) in romaine lettuce.

Four cultivars of lettuce, Lactuca sativa L., were evaluated for their resistance to the adult banded cucumber beetle, Diabrotica balteata LeConte, under laboratory conditions. When paired with each of the other three cultivars, leaf consumption in all possible combinations of short-term (48 h) two-choice tests among the cultivars was significantly reduced only for 'Valmaine'. However, in a 48-h no-choice situation, beetles fed Valmaine or 'Short Guzmaine' had similarly low leaf consumption, followed by 'Parris White', with the greatest consumption occurring on 'Tall Guzmaine'. In longer term experiments, female beetles fed Valmaine for 10, 13, or 16 d generally had lower survival and the lowest body weights compared with beetles fed any of the other three cultivars. No mature eggs were found in the ovaries of females fed Valmaine, whereas from 14% (Short Guzmaine, day 10) to 100% (Tall Guzmaine, day 13) of females fed the other cultivars produced mature eggs. In a starvation test, most D. balteata of either sex did not survive after 7 d with access only to water. Moreover, starved females did not produce mature eggs. Thus, food consumption by adult D. balteata is very important to their survival and reproductive performance, and it is likely that females fed Valmaine failed to produce mature eggs because they did not consume a sufficient amount of this cultivar. However, because Valmaine-fed beetles maintained their body weight and lived significantly longer than starved beetles, it appears that they can obtain some nourishment from their limited feeding on this cultivar. Overall, these results suggest that Valmaine, and to a lesser extent short Guzmaine (a cultivar produced by crossing Valmaine with two other cultivars), exhibit antixenosis-based resistance against D. balteata.

Impact of Bemisia argentifolii (Homoptera: Auchenorrhyncha: Aleyrodidae) infestation and squash silverleaf disorder on zucchini yield and quality.

Fruit yield and quality of zucchini, Cucurbita pepo L., plants infested with Bemisia argentifolii Bellows & Perring were evaluated in a screenhouse under spring and fall growing conditions by using closely related sister lines that were either susceptible (ZUC61) or tolerant (ZUC76-SLR) to squash silverleaf disorder. Our objective was to test separately the effects of level of whitefly infestation and expression of silverleaf symptoms on zucchini yield and quality. In a second experiment, yield and quality of fruit produced by silverleaf-tolerant zucchini genotypes incorporating two different sources of tolerance (ZUC76-SLR and ZUC33-SLR/PMR) were compared with that of 'Zucchini Elite', a silverleaf-susceptible commercial hybrid. Zucchini fruit yield was reduced in plants exposed to repeated infestations of whiteflies in spring and fall of both experiments. In addition, fruit grew to harvestable size more slowly under the highest whitefly infestations. Fruit quality was reduced at high infestations because of uneven and reduced pigmentation. The fruit yield and quality of ZUC61 and ZUC76-SLR were similarly affected by whitefly infestation despite differences in their susceptibility to squash silverleaf disorder. Fruit from infested plants showed decreased levels of chlorophyll and carotenoids causing the "blanching" of the fruit that is associated with loss of quality and reduced marketability. Leaves of infested plants of all genotypes had reduced levels of photosynthetic and photoprotectant pigments, possibly leading to reduced photosynthesis and consequently reduced yield. We conclude that feeding by high whitefly populations rather than expression of squash silverleaf disorder is responsible for yield and quality reduction in zucchini.

Arsenic hyperaccumulation in the Chinese brake fern (Pteris vittata) deters grasshopper (Schistocerca americana) herbivory.

Brake fern, Pteris vittata, not only tolerates arsenic but also hyperaccumulates it in the frond. The hypothesis that arsenic hyperaccumulation in this fern could function as a defense against insect herbivory was tested. Fronds from control and arsenic-treated ferns were presented to nymphs of the grasshopper Schistocerca americana. Feeding damage was recorded by visual observation and quantification of the fresh weight of frond left uneaten and number of fecal pellets produced over a 2-d period. Grasshopper weight was determined before and after 5 d of feeding. Grasshoppers consumed significantly greater amounts of the frond tissue, produced more fecal pellets and had increased body weight on control plants compared with grasshoppers fed arsenic-treated ferns. Very little or none of the arsenic-treated ferns were consumed indicating feeding deterrence. In a feeding deterrent experiment with lettuce, sodium arsenite at 1.0 mm deterred grasshoppers from feeding whereas 0.1 mm did not. In a choice experiment, grasshoppers preferred to feed on lettuce dipped in water compared with lettuce dipped in 1.0 mm sodium arsenite. Our results show that arsenic hyperaccumulation in brake fern is an elemental defense against grasshopper herbivory.