Insect feeding on cucumber mediated by rhizobacteria-induced plant resistance

Select strains of plant growth-promoting rhizobacteria (PGPR) were evaluated in greenhouse experiments with cucumber for induction of resistance against cucumber beetle (Diabrotica undecimpunctata howardi Barber) feeding and the beetle-transmitted cucurbit wilt disease. When beetles were given a choice between PGPR-treated and nontreated cucumber, their feeding on stems and cotyledons and the severity of wilt symptoms were significantly lower on PGPR-treated plants. HPLC analysis demonstrated that cotyledons from PGPR-treated plants contained significantly lower concentrations of the cucumber beetle feeding stimulant cucurbitacin than nontreated plants. These results suggest that a mechanism for PGPR-induced resistance against cucumber beetle feeding may involve a change in the metabolic pathway for cucurbitacin synthesis.     

Resistance of sweetpotato genotypes to adult Diabrotica beetles

Production of sweetpotatoes, Ipomoea batatas (L.) Lam. (Convolvulaceae), is limited by several insect pests, including Diabrotica spp. (Coleoptera: Chrysomelidae), and new integrated pest management (IPM) techniques for this crop are needed. Host plant resistance is one attractive approach that fits well into IPM programs. A host plant resistance research program typically depends on reliable bioassay procedures to streamline evaluation of germplasm. Thus, a bioassay technique was developed for evaluating sweetpotato germplasm by using adults of the banded cucumber beetle, Diabrotica balteata LeConte, and spotted cucumber beetle, Diabrotica undecimpunctata howardi Barber. A single beetle was placed on a piece of sweetpotato peel (periderm and cortex with stele removed) that was embedded periderm-side up in plaster in a petri dish. Feeding and longevity of insects on 30 sweetpotato genotypes were evaluated in two experiments by using this procedure. Adult longevity ranged from 7 to 11 d for starved individuals to 211 d for beetles fed a dry artificial diet. Longevity of banded cucumber beetles that fed on sweetpotato peels ranged from 12 d for the most-resistant genotype to 123 d for SC1149-19, a susceptible control cultivar. Longevity of spotted cucumber beetles was slightly shorter than longevity of banded cucumber beetles. For the most resistant sweetpotato genotypes, both Diabrotica species exhibited a significant delay in initiation of feeding, and more beetles died on these genotypes before they had fed. Both antibiosis and nonpreference (antixenosis) are important mechanisms of resistance in sweetpotato genotypes. This bioassay was consistent with field results, indicating that this technique could be useful for evaluating resistance to Diabrotica spp. in sweetpotato genotypes.     

Evaluation of cucurbitacin-based gustatory stimulant to facilitate cucumber beetle (Coleoptera: Chrysomelidae) management with foliar insecticides in melons

The bitter plant-derived compounds cucurbitacins are known to stimulate feeding of adult cucumber beetles (Coleoptera: Chrysomelidae). A cucurbitacin-based gustatory stimulant applied as a flowable bait combined with either spinosad or carbaryl was compared with foliar sprays of spinosad and carbaryl for controlling two cucumber beetle species (Diabrotica undecimpunctata undecimpunctata Mannerheim and Acalymma trivittatum Mannerheim) in honeydew melons (Cucumis melo L.). Field studies were conducted on the University of California-Davis plant pathology farm in 2008 and 2009. Beetle densities after applications and fruit damage from beetle feeding were compared among treatments. In addition, beetle survival was compared within field cages placed over the treated foliage infested with beetles. Using all three measures of efficacy, we determined that the addition of cucurbitacin bait had no effect on the level of cucumber beetle control with carbaryl in either 2008 or 2009. In both years, spinosad did not significantly reduce cucumber beetle densities in either field cages or field plots and did not reduce fruit damage relative to the untreated control. The addition of the bait to spinosad did not improve its efficacy. A laboratory bioassay of the spinosad formulation used in the field showed it had significant lethal effects on adults of both cucumber beetle species. Results indicated that the bait formulation used did not improve cucumber beetle control but may benefit from the addition of floral attractants or using a different type of cucurbitacin.     

Secondary metabolites as stimulants and antifeedants of Salix integra for the leaf beetle Plagiodera versicolora

Plagiodera versicolora, a willow beetle living on S. sachalinensis, is found on S. integra during early June in Hokkaido Island, Japan. This insect selects several species of willows (Salix), including S. integra as host plant in Honshu Island of Japan. To determine the reasons for the limited distribution of this beetle on the willows of Hokkaido, the feeding preference of the insect to leaves of S. integra and its constituents was performed. Feeding-bioassay guided fractionation of an 80% aqueous acetone extract of fresh leaves of Salix integra to Plagiodera versicolora resulted in isolation of feeding stimulant and antifeeding constituents. Chlorogenic acid (1) and 3,5-dicaffeoyl quinic acid (2) were identified as antifeedants and 1,2-di[(9Z,12Z,15Z)-octadeca-9,12,15-trienoyl]-3-beta-D-galactopyranosyl-sn-glycerol (MGDG, 3) as feeding stimulants. The feeding test was performed by an agar disk method. The treated agar disks contained sucrose and test sample in different doses. The antifeeding activities of 1 and 2 and stimulant activity of 3 may be one of the reasons for the limited presence of P. versicolora on S. integra in Hokkaido.     

Evaluation of phloxine B as a possible control agent against citrus thrips (Thysanoptera: Thripidae)

We evaluated the toxicity of phloxine B photoactive dye combined with a cane molasses bait against citrus thrips, Scirtothrips citri (Moulton). Laboratory bioassays conducted under artificial light showed that thrips mortality followed a log-dose probit-response model with an estimated LC50 of 0.0079% dye. Diluted cane molasses plus 0.01% phloxine B then was used as a standard for comparison of eight additional baits, including three formulations of concentrated citrus peel liquor (CCPL1-3). Citrus thrips mortality ranked highest to lowest with CCPL1 and CCPL3 > CCPL2, Mo-Bait and cane molasses > concentrated beet molasses, concentrated cane molasses, hemicellulose extract, and whey. Several commercial surfactants were tested to see if their addition to the standard increased efficacy. Hyper-Active, Kinetic, and Tween 60 at 1% and Cohort, Hyper-Active, Kinetic, and Silwet at 0.25% when added to the standard, reduced citrus thrips mortality, whereas Tween 60 and Dyne-Amic at 0.25% had no effect. Cane molasses with one or 5% phloxine B dye and CCPL1 with 1% dye were sprayed on citrus trees and allowed to weather in the field. Laboratory bioassays conducted after leaves had weathered for up to 8 d indicated that bait-dye toxicity was persistent. Possible use of the bait-dye mixture in commercial control of citrus thrips is discussed.     

Monitoring cucumber beetles in sweetpotato and cucurbits with kairomone-baited traps

Seven kairomone formulations (Trécé, Inc., Salinas, CA) were evaluated for their effectiveness as attractants for luring three species of cucumber beetles into Pherocon CRW traps (Trécé, Inc.) in cucurbit and sweetpotato fields. The spotted cucumber beetle, Diabrotica undecimpunctata howardi Barber; the banded cucumber beetle, Diabrotica balteata LeConte; and the striped cucumber beetle, Acalymma vittatum (F.), were captured in this study. TRE8276 (TIC mixture: 500 mg of 1,2,4-trimethoxybenzene, 500 mg of indole, and 500 mg of trans-cinnamaldeyde) and TRE8336 (500 mg of 1,2,4-trimethoxybenzene, 500 mg of trans-cinnamaldeyde, 500 mg of 4-methoxyphenethanol) were the most effective lures for spotted and striped cucumber beetles. None of the kairomone lures was very effective for attracting banded cucumber beetles. Three population peaks of spotted cucumber beetles were observed in cucurbit and sweetpotato fields at the U.S. Vegetable Laboratory (Charleston, SC). The efficacy of TRE8276 declined rapidly after 2 wk in the field. An improved design of the Pherocon CRW trap, with a yellow bottom and more-tapered top section, was more effective for capturing cucumber beetles than the original trap design made entirely of clear plastic. Banded cucumber beetles were not captured in sweetpotato fields at inland locations in North Carolina or South Carolina.     

Mutagenesis in cultured human diploid cells. IV. Induction of 8-azaguanine resistant mutations by phloxine, a mutagenic red dye.

Disodium 9-(3',4',5',6'-tetrachloro-o-carboxyphenyl)-6-hydroxy-2,4,5,7-tetrabromo-3-isoxanthone (phloxine), a red dye used as a food additive, was tested for its activity to induce 8-azaguanine (8AG) resistant mutations in cultured human embryonic cells. Phloxine had a severe cytotoxic effect on the cells at concentrations of 1 to 10 mug/ml. At concentrations of more than 30 mug/ml of phloxine no further decrease in cell survival was found. This cytotoxic effect of phloxine was not dependent on the duration of treatment. After treatment with phloxine for 2 h division of cells in normal medium was inhibited for 120 h. When cells were treated with phloxine at various concentrations for 2 h, cultured in normal medium for 48 h, and then selected with 30 mug/ml of 8AG, an increase in the induced mutation frequency was found. This increase in mutation frequency was dependent on the concentration of phloxine used as a mutagen and treatment with 100 mug/ml of phloxine increased the frequency to six times that in untreated cultures.     

Photoactive dye insecticide formulations: adjuvants increase toxicity to Mexican fruit fly (Diptera: Tephritidae)

The efficacy of photo active dyes as insecticides depends on the ingestion of the dye by the target insect and the activity of the dye at sensitive sites or on essential chemical functions. The site of this activity in insects is not understood, but we have found that certain chemical additives enhance the toxicity of phloxine B in the Mexican fruit fly. A series oftests with commercial adjuvants was performed under laboratory conditions that demonstrated a multifold decrease in the LD50 of phloxine B concentration and a decrease in the time required for photodynamic action to kill the flies. A total of 22 commercial adjuvants was tested. Of these, six were selected for evaluation under field cage conditions in comparison with a non-insecticide control bait (no treatment) and a phloxine B check bait with no adjuvant. Mortality was estimated by counting dead flies, feeding was estimated by fly counts at feeding stations, survival was estimated by trapping flies after the treatment period. In all cases the adjuvants increased the rate of mortality and decreased numbers surviving the treatment. Significant differences between adjuvants and both check and control were observed for mortality rates and the three best adjuvants, SM-9, Kinetic, and Tween60, induced significantly more mortality than the other adjuvants, the control, or the check. Feeding rates and survival rates indicated that the adjuvants increase the effectiveness of phloxine B in a predictable manner. The adjuvants appear to be active inside the insect rather than increasing the solubility of the dye in the bait medium. We propose that the addition of 1% vol:vol of the best adjuvant, Tween60 to the proteinaceous bait with 0.5% phloxine B will enhance toxicity as well as improve mixing and other characteristics of the bait.     

Attraction and feeding responses of Mediterranean fruit fly and a natural enemy to protein baits laced with two novel toxins,phloxine B and spinosada

Studies were conducted to determine attraction and feeding propensity of Mediterranean fruit fly, Ceratitis capitata (Wiedemann), to different protein bait mixtures with and without the insecticides malathion, spinosad, and phloxine B. Protein baits were more attractive to females than to males. Protein-starved females responded more than protein-fed females. The type of protein (USB^® yeast hydrolysate enzymatic, Mazoferm^®E802, Nu-Lure^®Insect Bait, or Provesta^® 621 autolyzed yeast extract) in the bait had a major influence on C. capitata attraction, which was strongest to fresh Provesta. Aged baits (four day-old) were not as attractive as fresh baits. In feeding propensity studies, highest response was observed for USB protein. On the basis of attraction and feeding responses Provesta (attraction and feeding) and USB (feeding) outperformed the standard Nu-Lure. Protein-starved flies were much more likely to feed on protein compared to protein-fed flies. For protein-starved flies, a mixture of Provesta and malathion repelled fruit flies, compared to a mixture of Provesta and spinosad or phloxine B. This was not the case with protein–fed flies. The wasp Fopius arisanus (Sonan), one of C. capitata's primary natural enemies in Hawaii, would not consume protein baits. Our studies suggest that spinosad or phloxine B, with low contact toxicity, mixed with protein baits offers a more environmentally friendly choice for control of C. capitata and conservation of F. arisanus, whereby the nontarget effects of broad spectrum contact poisons such as malathion can be avoided. Presumably, due to greater selectivity with spinosad and phloxine B bait treatments, the host would be killed, but not the natural enemy.     

Inhibition of seed germination by extracts of bitter Hawkesbury watermelon containing cucurbitacin, a feeding stimulant for corn rootworm (Coleoptera: Chrysomelidae)

Cucurbitacins are feeding stimulants for corn rootworm used in baits to control the adults of this insect pest. Corn rootworm larvae also feed compulsively on cucurbitacins. Cucurbitacins are reported to be gibberellin antagonists that may preclude their use as seed treatments for these soil-dwelling insects. The crude extract of a bitter Hawkesbury watermelon containing cucurbitacin E-glycoside significantly inhibited germination of watermelon, squash, and tomato seeds. Although the germination of corn seed was not significantly inhibited, root elongation was inhibited by crude extracts, but not by high-performance liquid chromatography-purified cucurbitacin E-glycoside. Therefore, the effects of the major components in the bitter watermelon extract (e.g., sugars) on seed germination and root elongation were determined. Pure sugars (glucose and fructose), at concentrations found in watermelon extract, mimicked the inhibition of seed germination and root elongation seen with the crude bitter Hawkesbury watermelon extract. Removal of these sugars may be necessary to use this extract as a bait for corn rootworm larvae as a seed or root treatment.     

Toxicity and residual effectiveness of insecticides on insecticide-treated spheres for controlling females of Rhagoletis pomonella (Diptera: Tephritidae)

This study evaluated the toxicity of five technical-grade insecticides of four different classes to apple maggot females, Rhagoletis pomonella (Walsh), following a 10-min exposure period in insecticide-coated glass jars, with or without a feeding stimulant (sucrose) present. According to LC90 values for toxicity by ingestion and tarsal contact, imidacloprid was 1.5 times more toxic than dimethoate or abamectin, diazinon was less toxic, and phloxine B (a phototoxic dye) least toxic. Based on LC90 values for tarsal contact alone, dimethoate was 2.3, 4.0, and 18.4 times more toxic than imidacloprid, abamectin, and diazinon, respectively. Contact alone with phloxine B caused no mortality. When exposure was assessed using spheres coated with a latex paint mixture containing sucrose and formulated dimethoate (Digon 400 EC) or imidacloprid (Provado 1.6 F) at concentrations ranging from 5 to 70 g (AI)/cm2, both insecticides showed reduced effectiveness compared with toxicities from glass jar tests, with Digon two times more toxic than Provado. After exposure to artificial rainfall and retreatment with sucrose, Digon- and Provado-treated spheres exhibited greatest residual effectiveness, with diazinon-treated spheres less effective. Spheres treated with formulated abamectin (Agri-Mek 0.15 EC) at 1.0% (AI) performed only slightly better than phloxine B-treated spheres, which completely lost effectiveness after exposure to rainfall. Spheres treated with formulated imidacloprid (Merit 75 WP) at 1.5% (AI) showed equal or better residual efficacy in killing apple maggot flies (> 80% mortality, shorter lethal duration of feeding) over a 12-wk exposure period to outdoor weather than spheres treated with Digon at 1.0% (AI) after both types were retreated with sucrose. Our results indicate that imidacloprid is a promising safe substitute for dimethoate as a fly killing agent on lure-kill spheres. Imidacloprid formulated as Merit 75 WP had greater residual efficacy than imidacloprid formulated as Provado 1.6 F.     

The effects of wilt symptom development and peroxidase induction on interactions between vascular wilt bacteria and cucumber beetles

Infection of cucumber (Cucumis sativus L.) with the bacterial pathogen Erwinia tracheiphila E. F. Smith causes vascular wilt disease in leaves, which may alter the suitability of the host plant for insects and other pathogens. In this study, densities of spotted (Diabrotica undecimpunctata howardi Barber) and striped (Acalymma vittata (Fab.) cucumber beetles (Coleoptera: Chrysomelidae) were higher on wilted leaves of mature and seedling field plants inoculated with E. tracheiphila. Bacterial infection or feeding by D. undecimpunctata howardii beetles increased total peroxidase enzyme activity in inoculated or infested leaves of greenhouse seedlings, but only beetle feeding induced higher activities in untreated systemic leaves on the same plants. Neither bacterial infection nor beetle infestation led to the development of systemic acquired resistance (SAR) to the fungal pathogen Colletotrichum orbiculare (Berk & Mont.) Arx. Susceptibility to this fungus was greater on E. tracheiphila-infected plants than on controls. The positive association between leaf wilt symptom development and beetle occurrence thus occurs in the presence of an oxidative but not anti-pathogenic response induced by both the insect and the pathogen.     

Field and vegetable crops as hosts of larval western spotted cucumber beetle (Coleoptera: Chrysomelidae)

The western spotted cucumber beetle, Diabrotica undecimpunctata undecimpunctata Mannerheim, is an important pest of melons (Cucurmis melo L.) in northern California. Recent observations indicate that adults are using alfalfa (Medicago sativa L.) as a feeding host and larvae may be developing on the roots. Greenhouse studies were conducted during the winters of 2009 and 2010 in which larval development was compared on the roots of six field and vegetable crops commonly grown in the southern Sacramento Valley. The growth parameters used to evaluate the hosts were larval and pupal head capsule width, body width, and body length as well as total survival percentage and survival percentage to the third instar. According to larval growth and survivorship in 2009, maize (Zea mays L.) was the best host, followed by alfalfa and tomato (Solanum lycopersicum L.) that were roughly equivalent to one another. Melon was a slightly weaker host than alfalfa and tomato; sunflower (Helianthus annuus L.) was a completely incompatible host and thus dropped from the 2010 study. In 2010, melon was the weakest host for larval development. Maize was the superior host again followed closely by alfalfa that performed slightly better than tomato and lima bean (Phaseolus lunatus L.). Data suggest that larval western spotted cucumber beetles may primarily develop outside of melon fields and the adults emigrate to melons.     

Consumptive and non‐consumptive effects of wolf spiders on cucumber beetles and cucumber plant damage

Biological control research often focuses on the ability of predators to reduce pest densities and protect crops through consumption. Less studied is their ability to protect crops by altering pest behaviour (non‐consumptive effects). Lab experiments were conducted to test predation rates of striped cucumber beetles (Acalymma vittatum; Coleoptera: Chrysomelidae) and spotted cucumber beetles (Diabrotica undecimpunctata howardi; Coleoptera: Chrysomelidae) by large (>10 mm) wolf spiders (Araneae: Lycosidae). Field experiments were conducted to examine how the physical presence and/or cues of spiders impact the behaviour and mortality of A. vittatum (specialist) and D. undecimpunctata (generalist) cucumber beetles as well as growth and damage of cucumber plants (Cucumis sativus; Cucurbitaceae). A. vittatum and D. undecimpunctata adults were added to caged cucumber plants without a spider, with spider cues only (spider removed before beetle inclusion), with spider only (spider introduced to plants immediately before beetle inclusion), and with spiders and their cues present (spiders introduced 24 hr in advance of beetle inclusion). A. vittatum responded to spider cues primarily by emigrating from plants. Contrarily, D. undecimpunctata did not display obvious responses, such as reduced feeding or increased emigration, to spider foraging and/or cues. Actively foraging lycosids increased A. vittatum mortality and reduced densities of D. undecimpunctata in the field when cucumber plants were flowering. This study highlights how non‐consumptive and consumptive effects can play a role in modifying pest populations, and how these effects can vary across species and plant growth stages.     

Watermelon juice: a promising feedstock supplement, diluent, and nitrogen supplement for ethanol biofuel production

Background  

Two economic factors make watermelon worthy of consideration as a feedstock for ethanol biofuel production. First, about 20% of each annual watermelon crop is left in the field because of surface blemishes or because they are misshapen; currently these are lost to growers as a source of revenue. Second, the neutraceutical value of lycopene and L-citrulline obtained from watermelon is at a threshold whereby watermelon could serve as starting material to extract and manufacture these products. Processing of watermelons to produce lycopene and L-citrulline, yields a waste stream of watermelon juice at the rate of over 500 L/t of watermelons. Since watermelon juice contains 7 to 10% (w/v) directly fermentable sugars and 15 to 35 μmol/ml of free amino acids, its potential as feedstock, diluent, and nitrogen supplement was investigated in fermentations to produce bioethanol.     

Insecticidal activity of photoactive dyes to American and migratory grasshoppers (Orthoptera: Acrididae)

Many photoactive dyes are relatively nontoxic to vertebrates despite their insecticidal properties. Several photoactive dyes known to be toxic to some groups of insects were evaluated at various concentrations for toxicity to American and migratory grasshoppers in laboratory and field studies. Rose bengal and phloxine B were effective at inducing mortality of grasshoppers when applied at 2 and 5% to bran bait, though erythrosin B and uranine were ineffective. Partial replacement of phloxine with uranine in dye mixtures resulted in no significant loss of efficacy. Some indication of feeding inhibition was observed at high dye concentrations, so minimum effective dosages, probably 2%, are optimal. Phloxine B and rose bengal appeared to be stable upon exposure to sunlight, and able to withstand at least 24 h of sunlight without significant degradation. Dyes such as phloxine B could be a viable grasshopper control agent for small or medium-sized grasshopper species because mortality can be induced by consumption of a single flake dusted with 5% dye, and yet pose little hazard to vertebrates. Large species such as American grasshopper must consume several flakes before mortality is induced.     

An alternative sampling technique for cucumber beetles (Coleoptera: Chrysomelidae) and diurnal beetle activity on muskmelon

Field studies were conducted to investigate the effectiveness of yellow sticky traps as an alternative sampling technique for striped, Acalymma vittatum (F.), and spotted, Diabrotica undecimpunctata howardi Barber, cucumber beetles (Coleoptera: Chrysomelidae) and the diurnal beetle activity on muskmelon, Cucumis melo L., near Vincennes, IN, in 2003 and 2004. The experimental design included six replications of seven 20-m-long rows each of muskmelon with 1.5 m between rows. On each sampling date, two yellow sticky traps were placed randomly between rows in each replication. One sticky trap was placed vertically with the lower edge even with the top of the canopy, whereas the other trap was placed horizontally, even with the top of the canopy. After traps were placed in the field, number of beetles on plants was counted in situ from 0800 to 1600 hours at 2-h intervals the next day. After 48 h in the field, the number of cucumber beetles adhering on traps was counted. Analyses of variance and Tukey's multiple comparison procedure were used to compare the densities of beetles among sampling times, and regression analyses were applied to correlate the numbers of beetles on traps and the numbers of in situ counts. Results show that both species of cucumber beetles were most active from 1200 to 1400 hours, and 20 beetles on the vertically positioned sticky trap were equivalent to one beetle per plant in the field. The application of the sampling technique and scouting time for cucumber beetle management are discussed.     

Romaine lettuce latex deters feeding of banded cucumber beetle: a vehicle for deployment of biochemical defenses

Latex is widely found among plant species and is known to play a defensive role against certain herbivores. Two romaine lettuce, Lactuca sativa L. (Asteraceae) cultivars, 'Valmaine' (resistant) and 'Tall Guzmaine' (susceptible), were selected to study the potential of latex as a defense mechanism against the banded cucumber beetle, Diabrotica balteata LeConte (Coleoptera: Chrysomelidae). Latex from Valmaine strongly inhibited feeding of adult D. balteata compared to latex from Tall Guzmaine when applied to the surface of artificial diet. Beetles consumed significantly less diet from disks treated with Valmaine latex than they consumed from diet treated with Tall Guzmaine latex, in both choice and no-choice tests. In a choice test involving diet disks treated with Valmaine latex from young leaves vs. disks treated with latex from mature leaves, the beetles consumed significantly less diet treated with latex from young leaves. No significant difference in diet consumption was found between diets treated with latex from young and mature leaves of Tall Guzmaine in choice tests. Three solvents of differing polarity (water, methanol, and methylene chloride) were tested to extract deterrent compounds from latex; Valmaine latex extracted with water:methanol (20:80) strongly inhibited beetle feeding when applied to the surface of artificial diet. In no-choice tests, fewer beetles were observed feeding on diet treated with water:methanol (20:80) extract of Valmaine latex than on diet treated with a similar extract of Tall Guzmaine latex, resulting in significantly less consumption of the diet treated with the Valmaine latex extract. These studies suggest that moderately polar chemicals within latex may account for resistance in Valmaine to D. balteata.     

Structure of the water-soluble feeding stimulant for Scolytus multistriatus: A revision

The feeding stimulant for the smaller European elm bark beetle has been revised to (+)-catechin 7-β-d-xylopyranoside (III) from the structure (I) bearing the sugar at the 5-position.