A PCR-based analysis of plant DNA reveals the feeding preferences of <Emphasis Type="Italic">Apolygus lucorum</Emphasis> (Heteroptera: Miridae)

The mirid bug Apolygus lucorum (Meyer-Dür) (Heteroptera: Miridae) is a severe pest of cotton and other crops in China. The feeding preferences of this pest are unclear due to its frequent movement among different host plants and the inconspicuous signs of its feeding. Here, we present results of a field trial that used direct observation of bug densities and a PCR-based molecular detection assay to detect plant DNA in bugs to explore relationships between A. lucorum population abundance and its feeding preference between two host plants, Humulus scandens (Loureiro) Merrill and Medicago sativa L. The field-plot samples showed that A. lucorum adults generally prefer flowering host plants. Its density was significantly higher on flowering H. scandens than on seedlings of M. sativa, and a similarly higher bug density was observed on flowering M. sativa than on seedlings of H. scandens. In the laboratory, we designed two pairs of species-specific primers targeting the trnL-F region for H. scandens and M. sativa, respectively. The detectability of plant DNA generally decreased with time post-feeding, and the half-life of plant DNA detection (DS₅₀) in the gut was estimated as 6.26 h for H. scandens and 3.79 h for M. sativa with significant differences between each other. In mirid bugs exposed to seedlings of H. scandens and flowering M. sativa, the detection rate of M. sativa DNA was significantly higher than that of H. scandens. Meanwhile, in mirid bugs exposed to seedlings of M. sativa and flowering H. scandens, a significantly higher detection rate of H. scandens DNA was found. We developed a useful tool to detect the remaining plant food species specifically from the gut of A. lucorum in the current study. We provided direct evidence of its feeding preference between H. scandens and M. sativa at different growth stages, which strongly supported a positive correlation between population abundance and feeding preference of A. lucorum on different plants under field conditions. The findings provide new insights into the understanding of A. lucorum’s feeding preference, and are helpful for developing the strategies to control this pest.     

Taxonomy and geographic distribution of <Emphasis Type="Italic">Carex lucorum</Emphasis> var. <Emphasis Type="Italic">austrolucorum</Emphasis> (section <Emphasis Type="Italic">Acrocystis</Emphasis>, Cyperaceae)

Carex lucorum currently comprises two morphologically similar varieties. We revise the taxonomy of C. lucorum, and map the distribution of the taxa. Based on multivariate analyses and analyses of variance of measured characters, allopatry, and previous studies revealing differences in achene micromorphology, chromosome differences, and flavonoid chemistry, we elevate C. lucorum var. austrolucorum to species status as C. austrolucorum. We provide an identification key to C. lucorum and its closest relatives.     

Host preferences and biotic potential of <Emphasis Type="Italic">Trialeurodes vaporariorum</Emphasis> and <Emphasis Type="Italic">Bemisia tabaci</Emphasis> (Hemiptera: Aleyrodidae) in tomato and pepper

Whiteflies Bemisia tabaci (Gennadius) and Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) are important pests in pepper (Capsicum annuum L.) and tomato (Solanum lycopersicum L.) crops in many countries. Contrary to what is observed for all other countries, in Uruguay, B. tabaci is mainly found on pepper and rarely on tomato, while T. vaporariorum is exclusively found on tomato. This study tested the oviposition preferences and biotic potential of these two whiteflies reared on both host plants. The developmental time, survival rates, longevity, fecundity and main population parameters were characterized. Both whitefly species showed different preference patterns regarding their host plants. T. vaporariorum preferred tomato instead of pepper to oviposit. Their developmental time is longer on pepper. B. tabaci preferred pepper, but the difference from tomato was not very strong. Pepper affects the biotic expression of T. vaporariorum negatively, while B. tabaci is able to develop equally on both host plants. These results show that the distribution differences of both whiteflies observed on both host plants could have a biological basis.     

An arthropod deterrent attracts specialised bees to their host plants

Many bee species are adapted to just a few specific plants in order to collect pollen (oligolecty). To reproduce successfully, it is important for oligolectic bees to find and recognise the specific host flowers. In this study, we investigated the role of floral volatiles used by an oligolectic bee to recognise its host plants. We compared the attractiveness of natural and synthetic scent samples of host flowers to foraging-naïve and -experienced Hoplitis adunca (Megachilidae) bees that are specialised on Echium and Pontechium (Boraginaceae) plants. The investigations showed that naïve H. adunca females are attracted to 1,4-benzoquinone. During their lifetime, bees learn additional floral cues while foraging on host flowers. In contrast to naïve ones, experienced H. adunca females use, in addition to 1,4-benzoquinone, other compounds to recognise their host plants. 1,4-Benzoquinone is an uncommon floral compound only known from the host plants of H. adunca, and is therefore ideally suited to be used as a plant-specific recognition cue. Several arthropods use this compound to deter insect predators. Therefore, 1,4-benzoquinone as an attractant in Echium flowers may have evolved from a primary function as a defensive compound against insect herbivores.     

Nymphal performance correlated with adult preference for flowering host plants in a polyphagous mirid bug, Apolygus lucorum (Heteroptera: Miridae)

Apolygus lucorum (Heteroptera: Miridae), a polyphagous mirid bug, often shows a strong preference for flowering host plants. It is hypothesized that host plant selection by phytophagous insects results in a variety of behavioral trade-offs. In this study, the relationship between adult preference and nymphal performance was examined on host plants with their flowers left intact and with flowers removed to investigate potential trade-offs in A. lucorum’s preference for flowering plants. Field trials in 2010 and 2011 showed that adults and nymphs were significantly more abundant on plants with flowers left intact than on those with flowers removed, while oviposition choice trials in field cages revealed that female adults preferred to lay eggs on plants with flowers. Laboratory performance trials demonstrated that adult lifespan and fecundity and nymphal development rate and survival were all significantly higher on plants with flowers compared with plants without flowers. In contrast, sex ratio was not significantly different. Simple linear regression analysis showed positive correlations between (1) adult and nymphal abundances in the field, (2) nymphal development rates and survival with adult fecundity in the laboratory, and (3) adult lifespan with fecundity in the laboratory. Since a positive preference–performance correlation was found for A. lucorum, we conclude that there are no evident behavioral trade-offs for host plant selection by this polyphagous mirid bug.     

Functional analysis of an <Emphasis Type="Italic">APETALA1</Emphasis>-like MADS box gene from <Emphasis Type="Italic">Eustoma grandiflorum</Emphasis> in regulating floral transition and formation

An Eustoma grandiflorum APETALA1 (EgAP1) gene showing high homology to the SQUA subfamily of MADS-box genes was isolated and characterized. EgAP1, containing a conserved euAP1 motif at the C-terminus, showed high sequence identity to Antirrhinum majus SQUAMOSA in the SQUA subfamily. EgAP1 mRNA was detected in the leaf and expressed significantly higher in young flower buds than in mature flower buds. In flowers, EgAP1 mRNA was strongly detected in sepal, weakly detected in petal and was absent in stamen and carpel. Transgenic Arabidopsis plants ectopically expressing EgAP1 flowered early and produced terminal flowers. In addition, the conversion of petals into stamen-like structures was also observed in 35S::EgAP1 flowers. 35S::EgAP1 was able to complement the ap1 flower defects by restoring the defect for sepal formation and significantly increasing second whorl petal production in Arabidopsis ap1 mutant plants. These results revealed that EgAP1 is the APETALA1 homolog in E. grandiflorum and that the function of EgAP1 is involved in floral induction and flower formation.     

On distribution and ecology of some species of cleonines (Coleoptera,Curculionidae): IV. Genus <Emphasis Type="Italic">Lixus</Emphasis> F., subgenus <Emphasis Type="Italic">Eulixus</Emphasis> Reitt.

Data on habitats, food plants of larvae and adults, feeding, mating, oviposition, larval and pupal development, natural enemies, and distribution of seven weevil species (Lixus canescens F.-W., L. iridis Ol., L. myagri Ol., L. punctirostris Boh., L. subtilis Boh., L. incanescens Boh., and L. brevipes Bris.) are given. New host plants of L. canescens and L. iridis are revealed. Gall induction by L. brevipes is reported for the first time. Distribution of all the species in Ukraine and Russia (the latter based on the literature) are given in more detail. Information on the known and potential economic importance of every species is provided.     

Impact of the invasive painted bug <Emphasis Type="Italic">Bagrada hilaris</Emphasis> on physiological traits of its host <Emphasis Type="Italic">Brassica oleracea</Emphasis> var botrytis

Bagrada hilaris is a herbivorous insect native of Asia and Africa, which has invaded southern Europe and North America where it causes major damage to cole crops. Laboratory experiments were conducted to assess how the infestation of this invasive species damages the host Brassica oleracea var botrytis, and to evaluate the interaction between plant emission of volatile organic compounds (VOC) and B. hilaris adults. Plant responses to insect feeding were evaluated through changes in photosynthesis, stomatal conductance, VOC emission, and visual damage on leaves. The impact of B. hilaris was compared with that of Nezara viridula, a polyphagous species distributed worldwide. Plant VOC role in host plant detection was tested with electroantennography bioassays on B. hilaris antenna. Photosynthesis and stomatal conductance were consistently reduced in plants infested with 40 B. hilaris adults for 24 h. The feeding activity of a single B. hilaris caused larger discolored spots on host leaves in comparison with N. viridula. VOC emitted by B. oleracea changed significantly in response to B. hilaris and N. viridula infestation. In particular, production of limonene was strongly reduced by the infestation of the two pentatomids, while an increase in the emission of acetic acid and 2-ethyl-1-hexanol was observed. EAG dose–response tests using the main plant VOC showed B. hilaris antennal responses to benzaldehyde, octanal, nonanal, and acetic acid, which indicates a role of these compounds in host location.     

Expression and DNA methylation pattern of reproduction-related genes in partially fertile triploid Pacific oysters <Emphasis Type="Italic">Crassostrea gigas</Emphasis>

Partial or complete sterility is an obvious feature in triploid Pacific oyster (Crassostrea gigas) which contributes to improving rearing performances. Despite the significance of sterility, the molecular mechanism behind it remains elusive and related research was limited. This study focused on six reproduction-related genes and compared their different behavior in gene expression and DNA methylation pattern between triploid and diploid oysters in order to provide more molecular information. The gonadal development of triploid oyster was examined by histology before molecular analysis. Gametogenesis disturbance was observed in triploid oysters at different development stages (stage II and III) with more serious impairment in females. QPCR showed significant gene expression difference between diploid and triploid in two genes: putative Vg and cgER. Gene expression of putative Vg was delayed in triploids while for cgER triploid oyster showed higher expression and the difference was significant at stage III. DNA methylation pattern of these two genes were further investigated by bisulfite sequencing. Between diploid and triploid oysters, no difference was observed in total methylation level but some individual loci showed different patterns: significantly high methylation rate of loci 2284 in cgER was observed in triploid oyster which has a higher expression of this gene. This study indicated that putative Vg and cgER might play a role in partial sterile in triploid C. gigas. Gene expression could be regulated by the methylation pattern at specific individual locus, which deserves equivalent attention as well as total DNA methylation level.     

Susceptibility of Asian citrus psyllid, <Emphasis Type="Italic">Diaphorina citri</Emphasis> (Hemiptera: Liviidae), to the insecticide afidopyropen: a new and potent modulator of insect transient receptor potential channels

A relatively new insecticide chemistry for controlling sucking insects, afidopyropen, was investigated for toxicity against Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). We evaluated the mortality of D. citri eggs, nymphs, and adults treated with afidopyropen using both laboratory-reared and field populations. We also quantified the effects of sublethal doses of afidopyropen on D. citri feeding, host choice selection, and fecundity. For laboratory susceptible adults, the contact LC₅₀, topical application LD₅₀, and leaf dip LC₅₀ were 2.13, 2.00, and 3.08 ng/µL, respectively. For adults collected from a commercially managed citrus grove in Florida, the contact LC₅₀, topical application LD₅₀, and leaf dip LC₅₀ were 1.37, 1.92, and 4.89 ng/µL, respectively. Egg hatch was significantly reduced following exposure to afidopyropen at 100 ng/µL. Furthermore, afidopyropen reduced D. citri nymph survival and adult emergence at concentrations ranging between 0.01 and 100 ng/µL. Diaphorina citri adult feeding decreased on citrus leaves treated with afidopyropen in a concentration-dependent manner as measured indirectly by honeydew excretion, and appeared almost completely inhibited after treatment with 10 and 100 ng/µL solutions of afidopyropen. In choice tests, significantly fewer D. citri adults settled on afidopyropen-treated plants than on control plants at 24, 48, and 72 h after release, with no differences in settling between males and females. Afidopyropen reduced the fecundity of D. citri in a concentration-dependent manner. Collectively, the results suggest that afidopyropen could contribute to the integrated management of D. citri and may therefore be useful in rotational programs to improve resistance management.     

The fitness of the cotton mealybug <Emphasis Type="Italic">Phenacoccus solenopsis</Emphasis> (Tinsley) is reduced after feeding on virus-infected <Emphasis Type="Italic">Hibiscus rosa</Emphasis>-<Emphasis Type="Italic">sinensis</Emphasis>

The cotton mealybug Phenacoccus solenopsis (Tinsley) and Cotton leaf curl Multan virus (CLCuMV), serious threats to economic crops and garden plants, have invaded southern China and widely infected Hibiscus rosa-sinensis. Whether an inter-species connection has facilitated the invasion process is unclear. In this study the interaction between P. solenopsis and H. rosa-sinensis infected with CLCuMV was investigated in the laboratory. We observed that 1st and 2nd instar nymphs of P. solenopsis preferred to feed on healthy H. rosa-sinensis leaves, whereas 3rd instar nymphs and female adults had no preference between healthy and virus-infected H. rosa-sinensis leaves. The developmental time of each P. solenopsis developmental stage increased significantly after feeding on infected H. rosa-sinensis leaves (p < 0.05). In particular, the development time for 2nd instar female and male nymphs and 3rd instar female nymphs increased by approximately twofold. The generation time of female mealybugs increased from 25.84 d on healthy H. rosa-sinensis to 32.12 d when feeding on CLCuMV-infected H. rosa-sinensis, and the survival rate decreased from 71.04 % on healthy H. rosa-sinensis to 5.80 % on infected plants. Nymph survival was most affected by feeding on infected plants. Additionally, the fecundity of female mealybugs feeding on infected H. rosa-sinensis decreased by 47.8 %. Thus, feeding on CLCuMV-infected H. rosa-sinensis significantly decreased the biological fitness and invading and colonizing abilities of P. solenopsis.     

Colonisation of primary and secondary host plant species by <Emphasis Type="Italic">Frankliniella schultzei</Emphasis> thrips: a balance between attraction and repulsion?

Common blossom thrips, Frankliniella schultzei Trybom (Thysanoptera: Thripidae), feed on pollen and flower tissues. They use multiple host species, but are regularly found in abundance on only some, with the red-flowered Malvaviscus arboreus the primary host in South-East Queensland. Flower-feeding insects commonly use plant odours and colour to recognise their usual hosts, so we quantified the attraction of flying thrips, in the field, to M. arboreus flowers relative to those of a secondary host (Hibiscus rosasinensis). More than two-thirds of the thrips were attracted to H. rosasinensis flowers over those of M. arboreus in a field test. We also compared flowers of these species in a cage, where significantly more thrips approached H. rosasinensis flowers than M. arboreus ones, and in an olfactometer, where significantly more approached the blank than flowers and leaves of either species. Thrips also avoided flower extracts in small arena-based studies. These thrips are clearly, but unexpectedly, more likely to approach the non-ancestral secondary host H. rosasinensis than their primary host plant M. arboreus (with which they presumably evolved), and are repelled by leaf and flower odours of both species. We propose that F. schultzei uses mainly vision in host recognition, and the hummingbird-pollinated M. arboreus has evolved insect-repellent properties. Residence times and oviposition rates in flowers across the two host species now warrant testing.     

The importance of resource distribution: spatial co-occurrence of host plants and host ants coincides with increased egg densities of the Dusky Large Blue <Emphasis Type="Italic">Maculinea nausithous</Emphasis> (Lepidoptera: Lycaenidae)

The occurrence of the Dusky Large Blue Butterfly (Maculinea nausithous) critically depends on the availability of two key resources: the Great Burnet (Sanguisorba officinalis) as primary nectar source for adults, for egg laying and early larval development, and the host ant Myrmica rubra as the food of late instar larvae. Thus, their distributions are key parameters shaping habitat suitability, and we expected that overlapping of both resources would have a strong impact on the size of local M. nausithous populations. Their egg density may be affected (a) by the fraction of host plants per site located within My. rubra activity ranges at the patch scale, or (b) by the availability of host plants with host ants in close range at the local scale, due to the potential ability of butterfly females to detect their host ants. To test the above hypothesis, we recorded spatial distribution patterns of host plants and host ants on 29 study sites in south-western Germany and related them to egg density data of M. nausithous. We found a positive relationship between co-occurence of host plant and host ant and M. nausithous egg density at the patch scale, whereas no correlation was found at the local scale. Thus, focal populations are strongly limited by the abundance of host plants, covered with My. rubra activity, as ant-mediated oviposition could not be proved. Our results underline the importance of resource distribution; the understanding of its impacts may provide useful insights into how M. nausithous habitats can be managed in order increase their carrying capacity.     

Reproductive Behavior of the Bug <Emphasis Type="Italic">Molipteryx fuliginosa</Emphasis> Uhler (Heteroptera,Coreidae) in the South of the Russian Far East

Reproductive behavior of Molipteryx fuliginosa (Uhler) was investigated in Primorskii Territory of Russia. From 4 to 18 repeated copulations of one female lasting from 2 to 48 hours were recorded in cages. The behavior of ovipositing females and the stages of oviposition are described for the first time. The number of eggs laid between copulations varied from 1 to 13, the number of oviposition acts, from 4 to 11, and the total female fecundity, from 21 to 38 eggs. Caged females laid eggs on plants and also on dead substrates unsuitable for nymphal feeding, such as cloth, dry branches, and a wooden pole. Copulation of M. fuliginosa was also observed under natural conditions. The preferred mating places of M. fuliginosa in anthropogenically modified habitats and in small-leaved riparian forests were plants of Rubus idaeus L., R. caesius L., and Rubus sp. After mating, females migrated in search of places for oviposition. Single eggs were found on the following plants not known previously as hosts of this bug: Solanum lycopersicum L., Carex sp., Elytrigia repens (L.) Nevski, and Taraxacum officinale Wigg. The females seemed to lack selectivity in the choice of place for oviposition, which was not always associated with host plants, despite their abundance and availability.     

Effects of alien invasion by <Emphasis Type="Italic">Bombus terrestris</Emphasis> L. (Apidae) on the visitation patterns of native bumblebees in coastal plants in northern Japan

When alien pollinator species enter a native community of pollinators in which resource partitioning has been established, the pollination network between plants and pollinators may be modified through the interactions between the pollinators over the use of floral resources. We observed the floral-use patterns of native (Bombus hypocrita and B. deuteronymus) and alien (B. terrestris) bumblebee species in a coastal grassland in northern Japan. We analyzed the factors determining resource partitioning patterns. B. hypocrita tended to visit flowers with shallow or wide open corollas, such as Rosa rugosa, whereas B. deuteronymus visited flowers with complex or deeper corollas, such as Lathyrus japonicus. Given the wider floral preference of B. terrestris, floral use by the alien bumblebees consistently overlapped with that of native bumblebees. The visitation of B. terrestris to R. rugosa flowers was positively correlated with that of B. hypocrita. These bumblebee species frequently used similar floral resources, in part because of the large overlap in the seasonality of their foraging activity. The visitation frequency of B. deuteronymus to L. japonicus flowers was independent of the visitation frequency of other bumblebee species. The major visitation periods of the bumblebees to L. japonicus flowers reciprocally differed between B. deuteronymus and B. terrestris, suggesting phenological resource partitioning between these species. Our study suggests that phenological niche partitioning is more common in specialized flowers (L. japonicus) than in generalized flowers (R. rugosa).     

<Emphasis Type="Italic">Agrobacterium rhizogenes</Emphasis> transformed calli of the holoparasitic plant <Emphasis Type="Italic">Phelipanche ramosa</Emphasis> maintain parasitic competence

Phelipanche and Orobanche spp. (broomrapes) are economically important parasitic weeds, causing severe damage to many agricultural crops. However, conventional methods to control these parasitic weeds are often not effective. Targeting molecular and biochemical processes involved in the establishment of the connection between the parasite and the host may offer a new perspective for control. However, progress in the understanding of these processes is hampered by the fact that genetic transformation and regeneration of these parasites is difficult if not impossible due to their specific lifecycle. Phelipanche and Orobanche spp. are holoparasites that need to attach to the roots of a host plant to get their assimilates, nutrients and water to develop and reproduce. The present study describes a highly efficient genetic transformation and regeneration protocol for the root holoparasitic Phelipanche ramosa. We present a new transformation system for P. ramosa using Agrobacterium rhizogenes MSU440 carrying a non-destructive selection marker gene coding for a red fluorescent protein (DsRed1). Using this protocol up to 90% transformation efficiency was obtained. We transformed 4 weeks old P. ramosa calli and transgenic calli expressing DsRed1 were then cultured on host plants. For the first time, we present shoot and flower development of the transgenic parasitic plant P. ramosa after successful connection of transgenic calli with the host plant roots. Moreover, we also present, for the first time, growth and development of P. ramosa shoots and flowers in vitro in the absence of a host plant.     

Reactive oxygen species (ROS) and antioxidative enzyme status in <Emphasis Type="Italic">Solanum lycopersicum</Emphasis> on priming with fluorescent <Emphasis Type="Italic">Pseudomonas</Emphasis> spp. against <Emphasis Type="Italic">Fusarium oxysporum</Emphasis>

In plants, ROS signaling and increase in activities of antioxidants are among defense responses. The present study describes the oxidative stress profiling in model host plant tomato (Solanum lycopersicum L.), during an invasion of the wilt pathogen Fusarium oxysporum f. sp. lycopersici with or without seed priming with Pseudomonas isolates M80, M96 and T109. Tomato seeds were primed with known Pseudomonas isolates M80, M96 and T109 and the forty-day- old plants were challenged with spores of F. oxysporum under greenhouse conditions. Leaf samples were collected at 0, 24, 48 72 and 96 h post fungal challenge and analysed for systemic level of oxidative stress parameters including total phenolics, proline, hydrogen peroxide, lipid peroxidation and enzymatic antioxidants. Disease incidence in the plants under greenhouse conditions was also calculated. Results revealed that priming with Pseudomonas isolates resulted in reduced oxidative stress in the host, during pathogen invasion. M80-priming showed highest antioxidative protection to the host plants during F. oxysporum invasion. The observed reduction in hydrogen peroxide and lipid peroxidation in primed plants was in agreement with the increased activities of the corresponding antioxidant enzymes. Greenhouse results showed that the highest wilt disease symptoms were with M80-priming followed by M96 and T109. The present study gives substantial evidences on the oxidative stress mitigation in response to Pseudomonas-priming on the model tomato-Fusarium interaction system.     

Soybean aphid, <Emphasis Type="Italic">Aphis glycines</Emphasis> (Hemiptera: Aphididae), developmental and reproductive capacity on white clover, <Emphasis Type="Italic">Trifolium repens</Emphasis> (Rosales: Leguminosae), in northeast China

Nymphs of Aphis glycines Matsumura were individually reared to adults in the laboratory on detached leaf discs of Trifolium repens L. (white clover) mounted on agar medium. Adults of A. glycines were fed T. repens within small clip cages in the field. Development, reproduction and intrinsic rates of increase of A. glycines were studied. These data were compared to those of controls fed known host plants including cultivated soybean Glycine max (L.) Merr. and the wild soybean species Glycine soja Sieb & Zucc. The results demonstrated that nymphs of A. glycines successfully developed into adults and reproduced efficiently when reared on T. repens in the laboratory. The lower development temperature threshold for nymphs fed T. repens was estimated as 8.27 °C, and the effective cumulative temperature for A. glycines development from nymph to adult was 90.91 degree-days. Adults of A. glycines could also survive on T. repens in the field, but only a few nymphs were produced.