A synchronized, large-scale field experiment using Arabidopsis thaliana reveals the significance of the UV-B photoreceptor UVR8 under natural conditions.

This study determines the functional role of the plant ultraviolet-B radiation (UV-B) photoreceptor, UV RESISTANCE LOCUS 8 (UVR8) under natural conditions using a large-scale 'synchronized-genetic-perturbation-field-experiment'. Laboratory experiments have demonstrated a role for UVR8 in UV-B responses but do not reflect the complexity of outdoor conditions where 'genotype × environment' interactions can mask laboratory-observed responses. Arabidopsis thaliana knockout mutant, uvr8-7, and the corresponding Wassilewskija wild type, were sown outdoors on the same date at 21 locations across Europe, ranging from 39°N to 67°N latitude. Growth and climatic data were monitored until bolting. At the onset of bolting, rosette size, dry weight, and phenolics and glucosinolates were quantified. The uvr8-7 mutant developed a larger rosette and contained less kaempferol glycosides, quercetin glycosides and hydroxycinnamic acid derivatives than the wild type across all locations, demonstrating a role for UVR8 under field conditions. UV effects on rosette size and kaempferol glycoside content were UVR8 dependent, but independent of latitude. In contrast, differences between wild type and uvr8-7 in total quercetin glycosides, and the quercetin-to-kaempferol ratio decreased with increasing latitude, that is, a more variable UV response. Thus, the large-scale synchronized approach applied demonstrates a location-dependent functional role of UVR8 under natural conditions.

Functionality of Selected Aromatic Lamiaceae in Attracting Pollinators in Central Spain.

Planting floral margins in agricultural landscapes has been shown to increase the abundance of pollinators in agro-ecosystems. However, to create efficient margins, it is necessary to use attractive, not weedy native plants with different blooming periods to prolong the availability of floral resources. Six native perennial plants of the Lamiaceae with different blooming periods were studied in a randomized block design, with the final aim to select the most efficient plants in floral mixtures by studying relationships between their floral phenology, floral density, and attractiveness to pollinators in Central Spain. In addition, their spatial expansion, i.e., potential weediness, was estimated under the field conditions, as the final purpose of the plants is to be implemented within the agro-ecosystems. The results showed that plant species with higher floral density (Nepeta tuberosa L. and Hyssopus officinalis L.) showed significantly higher attractiveness to pollinators and enhanced the attractiveness of floral mixtures. Species that bloomed in early spring (Salvia verbenaca L.) and in summer (Melissa officinalis L. and Thymbra capitata L.) did not efficiently contribute to the attractiveness of the mixtures to pollinators. In addition, besides high floral density of Salvia officinalis L. and N. tuberosa in the spring, warm and dry weather in spring 2012 enhanced the activity of bees, while cold and rainy weather in spring 2013 enhanced the activity of hoverflies. None of the plants showed weedy growth and so posed no danger of invading adjacent crops.

Metabolic detoxification of capsaicin by UDP-glycosyltransferase in three Helicoverpa species.

Capsaicin ß-glucoside was isolated from the feces of Helicoverpa armigera, Helicoverpa assulta, and Helicoverpa zea that fed on capsaicin-supplemented artificial diet. The chemical structure was identified by NMR spectroscopic analysis as well as by enzymatic hydrolysis. The excretion rates of the glucoside were different among the three species; those in the two generalists, H. armigera and H. zea, were higher than in a specialist, H. assulta. UDP-glycosyltransferases (UGT) enzyme activities measured from the whole larval homogenate of the three species with capsaicin and UDP-glucose as substrates were also higher in the two generalists. Compared among five different larval tissues (labial glands, testes from male larvae, midgut, the Malpighian tubules (MT), and fat body) from the three species, the formation of the capsaicin glucoside by one or more UGT is high in the fat body of all the three species as expected, as well as in H. assulta MT. Optimization of the enzyme assay method is also described in detail. Although the lower excretion rate of the unaltered capsaicin in H. assulta indicates higher metabolic capacity toward capsacin than in the other two generalists, the glucosylation per se seems to be insufficient to explain the decrease in capsaicin in the specialist, suggesting that H. assulta might have another important mechanism to deal with capsaicin more specifically.

Manipulating the attractiveness and suitability of hosts for diamondback moth (Lepidoptera: Plutellidae).

Ovipositional preference and larval survival of the diamondback moth, Plutella xylostella (L.), were compared among cabbage, Brassica oleracea L. variety capitata; glossy collards, Brassica oleracea L. variety acephala; and yellow rocket, Barbarea vulgaris (R. Br.) variety arcuata in different treatments of planting density, host plant age, intercropping, and water stress in 2003 and 2004. P. xylostella laid nearly twice as many eggs per plant in the high planting densities of glossy collards and yellow rocket than in the standard planting densities. Ovipositional preference was positively correlated with plant age in cabbage, glossy collards, and yellow rocket. Larval survival on cabbage was 1.9 times higher on 6-wk than on 12-wk-old plants, whereas larval survival on collards was 12.1 times higher on the younger plants. No larvae survived on either 6- or 12-wk-old yellow rocket plants. Intercropping cabbage with either tomato, Lycopersicon esculentum Mill., or fava bean, Vicia fava L., did not reduce the number of eggs laid on cabbage. No significant differences in oviposition were found between water-stressed and well-irrigated host plants treatments. Yet, P. xylostella larval survival on water-stressed cabbage was 2.1 times lower than on well-irrigated cabbage plants. Based on our findings, the effectiveness of trap crops of glossy collards and yellow rocket could be enhanced by integrating the use of higher planting densities in the trap crop than in the main crop and seeding of the trap crop earlier than the main crop.

Using yellow rocket as a trap crop for diamondback moth (Lepidoptera: Plutellidae).

Yellow rocket, Barbarea vulgaris (R. Br.) variety arcuata, was evaluated as a trap crop for diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), in cabbage, Brassica oleracea L. variety capitata, in 2003 and 2004. In 2003, the numbers of P. xylostella larvae found in field plots of cabbage alone were 5.2-11.3 times higher than those on cabbage plants in plots that included cabbage and several rows of yellow rocket. In an outdoor experiment in screenhouses, P. xylostella oviposition on cabbage was compared among six treatments that varied in the percentage of yellow rocket in relation to cabbage (0, 4, 8, 16, 24, and 32% of the plants were yellow rocket). Results indicated that the percentage of eggs laid on cabbage decreased as the percentage of yellow rocket in the treatment increased, but this decrease was not significant beyond 20% of the plants being yellow rocket. In 2004, the numbers of P. xylostella larvae in field plots of cabbage alone were 1.6-2.4 and 1.7-2.8 times higher than numbers in treatments with 10 and 20% trap crop, respectively. Sticky trap and sweep net captures of P. xylostella adults indicated that within-field dispersal was reduced by the presence of yellow rocket and aggregation occurred around yellow rocket plants. Our study suggests that using yellow rocket as a trap crop may reduce P. xylostella infestations in cabbage fields, and this possibility is discussed in the context of general crop and insect pest management practices in crucifers.

Evaluating trap crops for diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae).

Potential trap crops for the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), were evaluated through a series of ovipositional preference and larval survival experiments in outdoor screenhouses in 2002 and 2003. Hosts examined as trap crops were glossy and waxy collards, Brassica oleracea L. variety acephala; Indian mustard, Brassica juncea (L.) Czern; and yellow rocket, Barbarea vulgaris (R. Br.) variety arcuata. More eggs were laid on the potential trap crops, with the exception of waxy collards, than on cabbage. When P. xylostella was offered multiple hosts at the same time, numbers of eggs laid on glossy collards, Indian mustard, and yellow rocket were 3, 18, and 12 times greater than on cabbage, respectively. Similarly, when P. xylostella was offered a single trap crop host and cabbage, numbers of eggs laid on glossy collards, Indian mustard, and yellow rocket were 300, 19, and 110 times greater than on cabbage, respectively. Our studies suggest differences in oviposition between the potential trap crops and cabbage were likely due to host volatiles, leaf morphology and color, or a combination of these factors, rather than to total leaf areas, leaf shape, or plant architecture. Two-choice tests with a Y-tube olfactometer indicated that plant volatiles were major factors in P. xylostella host preference. The percentage larval survival from egg to pupation was 22.2% on cabbage, 18.9% on waxy collards, and 24.4% on Indian mustard, whereas survival was significantly lower on glossy collards (6.7%) and yellow rocket (0%). Based on our tests, it seems that yellow rocket may be the best candidate for use as a trap crop for P. xylostella because it is highly attractive for oviposition, but larvae do not survive on it.

Using plant chemistry and insect preference to study the potential of Barbarea (Brassicaceae) as a dead-end trap crop for diamondback moth (Lepidoptera: Plutellidae).

Barbarea vulgaris R. Br. has been proposed as a dead-end trap crop for diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), because its larvae do not survive on this plant species despite being highly preferred for oviposition. We compared plants of several species, varieties, and types in the genus Barbarea (Brassicaceae) to study their potential as trap crops for P. xylostella. In terms of insect behavior, Barbarea plants were assessed based on the criteria of high oviposition preference by P. xylostella moths (compared to other Barbarea plants and to three Brassica oleracea L. crop varieties) and low survival of P. xylostella larvae. Barbarea plants were also assessed based on the criteria of high content of glucosinolates, which stimulate adult oviposition and larval feeding in P. xylostella, and high content of saponins, which are detrimental to survival of P. xylostella larvae. All Barbarea plants tested were preferred over cabbage by ovipositing P. xylostella. Among Barbarea plants, few significant differences in oviposition preference by P. xylostella were found. Ovipositing P. xylostella preferred B. vulgaris plants containing mainly 2-phenylethylglucosinolate over B. vulgaris plants containing mainly (S)-2-hydroxy-2-phenylethylglucosinolate, and P-type B. vulgaris var. arcuata plants over Barbarea rupicola and B. vulgaris var. variegata plants. Despite containing a lower content of saponins than other Barbarea plants tested, Barbarea verna did not allow survival of P. xylostella larvae. Our studies show that, except for B. rupicola and P-type B. vulgaris var. arcuata, which allowed survival of P. xylostella larvae, all Barbarea plants tested have potential as dead-end trap crops for P. xylostella.

Damage to Miconia calvescens and seasonal abundance of Salbia lotanalis (Lepidoptera: Crambidae) in Costa Rica.

Miconia calvescens de Candolle (Melastomataceae) is an invasive tree considered the most serious threat to natural ecosystems of Hawaii and other Pacific islands. The success of M. calvescens as an invasive species is greatly owing to its shade tolerance and the shaded habitat it creates, where many native plant species that are light-demanding cannot survive. Salbia lotanalis Druce (Lepidoptera: Crambidae), a neotropical leaf roller attacking M. calvescens, was evaluated for two mechanisms by which it reduces leaf area of its host plant: feeding (defoliation), which removes leaf tissue, and tying leaf rolls, which reduces exposed area of leaves. These impacts were quantified over a 1-yr period at a field site in Costa Rica, where densities of S. lotanalis larvae attacking M. calvescens peaked at the end of the rainy season and declined in the dry season. Up to 47.5% of leaves were attacked by S. lotanalis, with cumulative defoliation by an undetermined number of larvae removing an average of ≍30% (253 cm(2)) of each leaf attacked. Defoliation and leaf rolling were compared in a greenhouse experiment in which individual S. lotanalis larvae defoliated an average of 3.7% (17.8 cm(2)) of each attacked leaf, and reduced exposed leaf area as a result of leaf rolling by an average of 12.8% (66.2 cm(2)). Our results complement the findings of previous studies of S. lotanalis and confirm its potential as a biological control agent of M. calvescens.

Insect attraction versus plant defense: young leaves high in glucosinolates stimulate oviposition by a specialist herbivore despite poor larval survival due to high saponin content.

Glucosinolates are plant secondary metabolites used in plant defense. For insects specialized on Brassicaceae, such as the diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), glucosinolates act as "fingerprints" that are essential in host plant recognition. Some plants in the genus Barbarea (Brassicaceae) contain, besides glucosinolates, saponins that act as feeding deterrents for P. xylostella larvae, preventing their survival on the plant. Two-choice oviposition tests were conducted to study the preference of P. xylostella among Barbarea leaves of different size within the same plant. P. xylostella laid more eggs per leaf area on younger leaves compared to older ones. Higher concentrations of glucosinolates and saponins were found in younger leaves than in older ones. In 4-week-old plants, saponins were present in true leaves, while cotyledons contained little or no saponins. When analyzing the whole foliage of the plant, the content of glucosinolates and saponins also varied significantly in comparisons among plants that were 4, 8, and 12 weeks old. In Barbarea plants and leaves of different ages, there was a positive correlation between glucosinolate and saponin levels. This research shows that, in Barbarea plants, ontogenetical changes in glucosinolate and saponin content affect both attraction and resistance to P. xylostella. Co-occurrence of a high content of glucosinolates and saponins in the Barbarea leaves that are most valuable for the plant, but are also the most attractive to P. xylostella, provides protection against this specialist herbivore, which oviposition behavior on Barbarea seems to be an evolutionary mistake.

Interaction of glucosinolate content of Arabidopsis thaliana mutant lines and feeding and oviposition by generalist and specialist lepidopterans.

The diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), is an insect specialized on glucosinolate-containing Brassicaceae that uses glucosinolates in host-plant recognition. We used wild-type and mutants of Arabidopsis thaliana (L.) Heynh. (Brassicaceae) to investigate the interaction between plant glucosinolate and myrosinase content and herbivory by larvae of the generalist Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) and the specialist P. xylostella. We also measured glucosinolate changes as a result of herbivory by these larvae to investigate whether herbivory and glucosinolate induction had an effect on oviposition preference by P. xylostella. Feeding by H. armigera and P. xylostella larvae was 2.1 and 2.5 times less, respectively, on apk1 apk2 plants (with almost no aliphatic glucosinolates) than on wild-type plants. However, there were no differences in feeding by H. armigera and P. xylostella larvae on wild-type, gsm1 (different concentrations of aliphatic glucosinolates compared to wild-type plants), and tgg1 tgg2 plants (lacking major myrosinases). Glucosinolate induction (up to twofold) as a result of herbivory occurred in some cases, depending on both the plant line and the herbivore. For H. armigera, induction, when observed, was noted mostly for indolic glucosinolates, while for P. xylostella, induction was observed in both aliphatic and indolic glucosinolates, but not in all plant lines. For H. armigera, glucosinolate induction, when observed, resulted in an increase of glucosinolate content, while for P. xylostella, induction resulted in both a decrease and an increase in glucosinolate content. Two-choice tests with wild-type and mutant plants were conducted with larvae and ovipositing moths. There were no significant differences in preference of larvae and ovipositing moths between wild-type and gsm1 mutants and between wild-type and tgg1 tgg2 mutants. However, both larvae and ovipositing moths preferred wild-type over apk1 apk2 mutants. Two-choice oviposition tests were also conducted with P. xylostella moths comparing undamaged plants to plants being attacked by larvae of either P. xylostella or H. armigera. Oviposition preference by P. xylostella was unaffected as a result of larval plant damage, even in the cases where herbivory resulted in glucosinolate induction.

A host-plant specialist, Helicoverpa assulta, is more tolerant to capsaicin from Capsicum annuum than other noctuid species.

Plant secondary compounds not only play an important role in plant defense, but have been a driving force for host adaptation by herbivores. Capsaicin (8-methyl-N-vanillyl-6-nonenamide), an alkaloid found in the fruit of Capsicum spp. (Solanaceae), is responsible for the pungency of hot pepper fruits and is unique to the genus. The oriental tobacco budworm, Helicoverpa assulta (Lepidoptera: Noctuidae), is a specialist herbivore feeding on solanaceous plants including Capsicum annuum, and is one of a very few insect herbivores worldwide capable of feeding on hot pepper fruits. To determine whether this is due in part to an increased physiological tolerance of capsaicin, we compared H. assulta with another specialist on Solanaceae, Heliothis subflexa, and four generalist species, Spodoptera frugiperda, Heliothis virescens, Helicoverpa armigera, and Helicoverpa zea, all belonging to the family Noctuidae. When larvae were fed capsaicin-spiked artificial diet for the entire larval period, larval mortality increased in H. subflexa and H. zea but decreased in H. assulta. Larval growth decreased on the capsaicin-spiked diet in four of the species, was unaffected in H. armigera and increased in H. assulta. Food consumption and utilization experiments showed that capsaicin decreased relative consumption rate (RCR), relative growth rate (RGR) and approximate digestibility (AD) in H. zea, and increased AD and the efficiency of conversion of ingested food (ECI) in H. armigera; whereas it did not significantly change any of these nutritional indices in H. assulta. The acute toxicity of capsaicin measured by injection into early fifth instar larvae was less in H. assulta than in H. armigera and H. zea. Injection of high concentrations produced abdominal paralysis and self-cannibalism. Injection of sub-lethal doses of capsaicin resulted in reduced pupal weights in H. armigera and H. zea, but not in H. assulta. The results indicate that H. assulta is more tolerant to capsaicin than the other insects tested, suggesting that this has facilitated expansion of its host range within Solanaceae to Capsicum after introduction of the latter to the Old World about 500 years ago. The increased larval survival and growth due to chronic dietary exposure to capsaicin suggests further adaptation of H. assulta to that compound, the mechanisms of which remain to be investigated.

Parallel evolution of Bacillus thuringiensis toxin resistance in lepidoptera.

Despite the prominent and worldwide use of Bacillus thuringiensis (Bt) insecticidal toxins in agriculture, knowledge of the mechanism by which they kill pests remains incomplete. Here we report genetic mapping of a membrane transporter (ABCC2) to a locus controlling Bt Cry1Ac toxin resistance in two lepidopterans, implying that this protein plays a critical role in Bt function.

Can sulfur fertilisation improve the effectiveness of trap crops for diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae)?

BACKGROUND: The effect of sulfur fertilisation on chemical constituents of yellow rocket, Barbarea vulgaris (R. Br.), was studied with regard to its potential use as a trap crop for the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae). Two types of B. vulgaris var. arcuata were used: the G-type, resistant to P. xylostella and proposed as a 'dead-end' trap crop, and the P-type, not resistant to P. xylostella and used as a control. RESULTS: In G-type B. vulgaris, sulfur fertilisation increased the content of the dominant glucosinolate (S)-2-hydroxy-2-phenylethylglucosinolate by 20%. Sulfur fertilisation did not significantly change levels of the saponins 3-0-beta-cellobiosylhederagenin and 3-0-beta-cellobiosyloleanolic acid, known to act as feeding deterrents for P. xylostella larvae. In P-type B. vulgaris, the same levels of sulfur fertilisation did not change the glucosinolate content significantly. Two-choice oviposition preference tests with B. vulgaris plants showed that P. xylostella laid 144% and 45% more eggs on G- and P-type plants with sulfur fertilisation respectively. CONCLUSIONS: The studies suggest that sulfur fertilisation could increase the effectiveness of G-type B. vulgaris as a trap crop for P. xylostella. The effect of plant sulfur fertilisation on P. xylostella oviposition preference is associated with a quantitative glucosinolate increase, but other compounds could also be involved.