Hcp2, a secreted protein of the phytopathogen Pseudomonas syringae pv. tomato DC3000, is required for competitive fitness against bacteria and yeasts

When analysing the secretome of the plant pathogen Pseudomonas syringae pv. tomato (Pst) DC3000, we identified hemolysin co-regulated protein (Hcp) as one of the secreted proteins. Hcp is assumed to be an extracellular component of the type VI secretion system (T6SS). Two copies of hcp genes are present in the Pst DC3000 genome, hcp1 (PSPTO_2539) and hcp2 (PSPTO_5435). We studied the expression patterns of hcp genes and tested the fitness of hcp knock-out mutants in host plant colonization and in inter-microbial competition. We found that the hcp2 gene is expressed, most actively at the stationary growth phase, and that the Hcp2 protein is secreted via T6SS and appears in the culture medium as covalently linked dimers. Expression of hcp2 is not induced in planta and it does not contribute to virulence or colonisation in tomato or Arabidopsis plants. Instead, hcp2 is required for survival in competition with enterobacteria and yeasts, and its function is associated with suppression of the growth of these competitors. This is the first report on bacterial T6SS-associated genes functioning in competition against yeast. Our results suggest that the T6SS of P. syringae may play an important role in bacterial fitness, allowing this plant pathogen to survive in conditions where it has to compete with other micro-organisms for resources.

Thorn-dwelling ants provide antiherbivore defence for camelthorn trees, Vachellia erioloba, in Namibia

Ants are widely employed by plants as an antiherbivore defence. A single host plant can associate with multiple, symbiotic ant species, although usually only a single ant species at a time. Different plant-ant species may vary in the degree to which they defend their host plant. In Kenya, ant–acacia interactions are well studied, but less is known about systems elsewhere in Africa. A southern African species, Vachellia erioloba, is occupied by thorn-dwelling ants from three different genera. Unusually, multiple colonies of all these ants simultaneously and stably inhabit trees. We investigated if the ants on V. erioloba (i) deter insect herbivores; (ii) differ in their effectiveness depending on the identity of the herbivore; and (iii) protect the tree against an important herbivore, the larvae of the lepidopteran Gonometa postica. We show that experimental exclusion of ants leads to greater levels of herbivory on trees. The ants inhabiting V. erioloba are an effective deterrent against hemipteran and coleopteran, but not lepidopteran herbivores. Defensive services do not vary among ant species, but only Crematogaster ants exhibit aggression towards G. postica. This highlights the potential of the V. erioloba–ant mutualism for studying ant–plant interactions that involve multiple, simultaneously resident thorn-dwelling ant species.

Molecular species delimitation of a symbiotic fig-pollinating wasp species complex reveals extreme deviation from reciprocal partner specificity

Background: Symbiotic relationships have contributed to major evolutionary innovations, the maintenance of fundamental ecosystem functions, and the generation and maintenance of biodiversity. However, the exact nature of host/symbiont associations, which has important consequences for their dynamics, is often poorly known due to limited understanding of symbiont taxonomy and species diversity. Among classical symbioses, figs and their pollinating wasps constitute a highly diverse keystone resource in tropical forest and savannah environments. Historically, they were considered to exemplify extreme reciprocal partner specificity (one-to-one host-symbiont species relationships), but recent work has revealed several more complex cases. However, there is a striking lack of studies with the specific aims of assessing symbiont diversity and how this varies across the geographic range of the host. Results: Here, we use molecular methods to investigate cryptic diversity in the pollinating wasps of a widespread Australian fig species. Standard barcoding genes and methods were not conclusive, but incorporation of phylogenetic analyses and a recently developed nuclear barcoding gene (ITS2), gave strong support for five pollinator species. Each pollinator species was most common in a different geographic region, emphasising the importance of wide geographic sampling to uncover diversity, and the scope for divergence in coevolutionary trajectories across the host plant range. In addition, most regions had multiple coexisting pollinators, raising the question of how they coexist in apparently similar or identical resource niches. Conclusion: Our study offers a striking example of extreme deviation from reciprocal partner specificity over the full geographical range of a fig-wasp system. It also suggests that superficially identical species may be able to co-exist in a mutualistic setting albeit at different frequencies in relation to their fig host’s range. We show that comprehensive sampling and molecular taxonomic techniques may be required to uncover the true structure of cryptic biodiversity underpinning intimate ecological interactions.

Tales of the unexpected: research discoveries that occurred by accident

Data from four experimental research projects are presented which have in common that unexpected results caused a change in direction of the research. A plant growth accelerator caused the appearance of white black bean aphids, a synthetic pyrethroid suspected of enhancing aphid reproduction proved to enhance plant growth, a chance conversation with a colleague initiated a search for fungal DNA in aphids, and the accidental invasion of aphid cultures by a parasitoid reversed the aphid population ranking of two Brussels sprout cultivars. This last result led to a whole series of studies on the plant odour preferences of emerging parasitoids which in turn revealed the unexpected phenomenon that chemical cues to the maternal host plant are left with the eggs at oviposition. It is pointed out that, too often, researchers fail to follow up unexpected results because they resist accepting flaws in their hypotheses; also that current application criteria for research funding make it hard to accommodate unexpected findings.

The diversity of arbuscular mycorrhizas of selected Australian Fabaceae

Members of the Australian native perennial Fabaceae have been little explored with regard to their root biology and the role played by arbuscular mycorrhizal (AM) fungi in their establishment, nutrition and long-term health. The ultimate goal of our research is to determine the dependency of native perennial legumes on their co-evolved AM fungi and conversely, the impact of AM fungal species in agricultural fields on the productivity of sown native perennial legume pastures. In this paper we investigate the colonisation morphology in roots and the AMF, identified by spores extracted from rhizosphere soil, from three replicate plots of each of the native legumes, Cullen australasicum, C. tenax and Lotus australis and the exotic legumes L. pedunculatus and Medicago sativa. The plants were grown in an agricultural field. The level and density of colonisation by AM fungi, and the frequency of intraradical and extraradical hyphae, arbuscules, intraradical spores and hyphal coils all differed between host plants and did not consistently differ between native and exotic species. However, there were strong similarities between species in the same genus. The three dominant species of AM fungi in rhizosphere soil also differed with host plant, but one fungus (Glomus mosseae) was always the most dominant. Sub-dominant AM species were the same between species in the same genus. No consistent differences in dominant spores were observed between the exotic and native Fabaceae species. Our results suggest that plant host influences the mycorrhizal community in the rhizosphere soil and that structural and functional differences in the symbiosis may occur at the plant genus level, not the species level or due to provenance.

Long-term changes to the frequency of occurrence of British moths are consistent with opposing and synergistic effects of climate and land-use changes

1. Species’ distributions are likely to be affected by a combination of environmental drivers. We used a data set of 11 million species occurrence records over the period 1970–2010 to assess changes in the frequency of occurrence of 673 macro-moth species in Great Britain. Groups of species with different predicted sensitivities showed divergent trends, which we interpret in the context of land-use and climatic changes. 2. A diversity of responses was revealed: 260 moth species declined significantly, whereas 160 increased significantly. Overall, frequencies of occurrence declined, mirroring trends in less species-rich, yet more intensively studied taxa. 3. Geographically widespread species, which were predicted to be more sensitive to land use than to climate change, declined significantly in southern Britain, where the cover of urban and arable land has increased. 4. Moths associated with low nitrogen and open environments (based on their larval host plant characteristics) declined most strongly, which is also consistent with a land-use change explanation. 5. Some moths that reach their northern (leading edge) range limit in southern Britain increased, whereas species restricted to northern Britain (trailing edge) declined significantly, consistent with a climate change explanation. 6. Not all species of a given type behaved similarly, suggesting that complex interactions between species’ attributes and different combinations of environmental drivers determine frequency of occurrence changes. 7. Synthesis and applications. Our findings are consistent with large-scale responses to climatic and land-use changes, with some species increasing and others decreasing. We suggest that land-use change (e.g. habitat loss, nitrogen deposition) and climate change are both major drivers of moth biodiversity change, acting independently and in combination. Importantly, the diverse responses revealed in this species-rich taxon show that multifaceted conservation strategies are needed to minimize negative biodiversity impacts of multiple environmental changes. We suggest that habitat protection, management and ecological restoration can mitigate combined impacts of land-use change and climate change by providing environments that are suitable for existing populations and also enable species to shift their ranges.

Nitrogen fluxes from treefrogs to tank epiphytic bromeliads: an isotopic and physiological approach

Diverse invertebrate and vertebrate species live in association with plants of the large Neotropical family Bromeliaceae. Although previous studies have assumed that debris of associated organisms improves plant nutrition, so far little evidence supports this assumption. In this study we used isotopic ((15)N) and physiological methods to investigate if the treefrog Scinax hayii, which uses the tank epiphytic bromeliad Vriesea bituminosa as a diurnal shelter, contributes to host plant nutrition. In the field, bromeliads with frogs had higher stable N isotopic composition (delta(15)N) values than those without frogs. Similar results were obtained from a controlled greenhouse experiment. Linear mixing models showed that frog feces and dead termites used to simulate insects that eventually fall inside the bromeliad tank contributed, respectively, 27.7% (+/- 0.07 SE) and 49.6% (+/- 0.50 SE) of the total N of V. bituminosa. Net photosynthetic rate was higher in plants that received feces and termites than in controls; however, this effect was only detected in the rainy, but not in the dry season. These results demonstrate for the first time that vertebrates contribute to bromeliad nutrition, and that this benefit is seasonally restricted. Since amphibian-bromeliad associations occur in diverse habitats in South and Central America, this mechanism for deriving nutrients may be important in bromeliad systems throughout the Neotropics.

Resource defense polygyny shifts to female defense polygyny over the course of the reproductive season of a Neotropical harvestman

Although studies classify the polygynous mating system of a given species into female defense polygyny (FDP) or resource defense polygyny (RDP), the boundary between these two categories is often slight. Males of some species may even shift between these two types of polygyny in response to temporal variation in social and environmental conditions. Here, we examine the mating system of the Neotropical harvestman Acutisoma proximum and, in order to assess if mate acquisition in males corresponds to FDP or RDP, we tested four contrasting predictions derived from the mating system theory. At the beginning of the reproductive season, males fight with other males for the possession of territories on the vegetation where females will later oviposit, as expected in RDP. Females present a marked preference for specific host plant species, and males establish their territories in areas where these host plants are specially abundant, which is also expected in RDP. Later in the reproductive season, males reduce their patrolling activity and focus on defending individual females that are ovipositing inside their territories, as what occurs in FDP. This is the first described case of an arachnid that exhibits a shift in mating system over the reproductive season, revealing that we should be cautious when defining the mating system of a species based on few observations concentrated in a brief period.

Resistência de genótipos de batata (Solanum spp.) a Diabrotica speciosa (Germar, 1824) (Coleoptera: Chrysomelidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Preferência alimentar de Dione juno juno (Cramer) por genótipos de maracujazeiro e avaliação do uso de extratos aquosos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Preferência para oviposição de Bemisia tabaci (Genn.) biótipo B (Hemiptera: Aleyrodidae) em genótipos de soja, sob condições de campo

Dez genótipos de soja: IAC 100, IAC 74-2832, IAC 90-2971, IAC 78-2318, PI 227687, PI 229358, PI 274454, BR-82 12547, MG/BRS-68 (Vencedora) e FT-5 (Formosa), foram avaliados quanto a resistência, do tipo não-preferência para oviposição, à mosca-branca, Bemisia tabaci (Genn.) biótipo B. O ensaio foi realizado no campo experimental da FCAV/UNESP, Jaboticabal-SP, de outubro de 1999 a janeiro de 2000. Pretendeu-se, ainda, verificar a preferência de oviposição da referida espécie em relação às folhas unifolioladas e trifolioladas, e quanto à posição do folíolo na folha trifoliolada, e estabelecer o número mínimo de folíolos necessários para amostrar adequadamente a oviposição da mosca branca. Constatou-se que a oviposição foi menor nos genótipos BR-82 12547 e IAC 74-2832 em todas as avaliações (15, 30, 45, 65 e 85 dias após a emergência das plantas). As maiores taxas de oviposição ocorreram em IAC 90-2971, IAC 100, PI 227687 e PI 274454. O número médio de ovos depositados por seção de folha pelas fêmeas de B. tabaci biótipo B nas folhas trifolioladas não diferiu significativamente dos obtidos nas unifolioladas, 15 dias após a emergência das plantas (estádios de desenvolvimento entre V3 e V5), em 70% dos genótipos. Quanto à posição dos folíolos, a mosca-branca ovipositou indistintamente sobre os folíolos mediano e laterais. Também o número médio de ovos depositados por área foliar, pela mosca-branca, nos três folíolos da folha trifoliolada não diferiu significativamente da média obtida para dois ou um folíolo desta, o que sugere ser, um folíolo, uma unidade amostral representativa da oviposição de B. tabaci biótipo B em soja.

FLUTUAÇÃO POPULACIONAL DE Dione juno juno (Cramer, 1779) (Lepidoptera: Nymphalidae) em MARACUJAZEIROS (Passiflora spp.), MÉTODOS DE AMOSTRAGEM E RESISTÊNCIA DE GENÓTIPOS

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Feeding non-preference by Spodoptera frugiperda and Spodoptera eridania on tomato genotypes

Lagartas do gênero Spodoptera spp. são altamente polífagas, podendo causar danos econômicos em diversas culturas agrícolas. em vista de sua emergente importância na cultura do tomate, principalmente o destinado à indústria, este trabalho teve por objetivo avaliar a não preferência, para alimentação, de lagartas de Spodoptera frugiperda (J. E. Smith, 1797) e Spodoptera eridania (Cramer, 1782) por genótipos de tomateiro, e classificá-los quanto aos graus de resistência. Como padrão susceptível, utilizou-se o cultivar comercial Santa Clara e, como resistente, a linhagem PI 134417, sendo avaliadas, ainda, as linhagens PI 134418, PI 126931, LA 462 e LA 716. Realizaram-se testes de não preferência, para alimentação, com e sem chance de escolha, avaliando-se a atratividade dos genótipos de tomateiro para as lagartas, em tempos pré-estabelecidos após sua liberação, além da massa foliar consumida. em geral, os genótipos LA 716 e PI 126931 foram os menos atrativos para a S. frugiperda, enquanto Santa Clara foi o mais atrativo e consumido. Quanto a S. eridania, os genótipos PI 126931, LA 462, LA 716 e PI 134418 foram os menos preferidos, para a alimentação, pelas lagartas, e Santa Clara e PI 134417 foram os mais atrativos e consumidos. Os genótipos LA e PI 126931 são moderadamente resistentes, do tipo não preferência para alimentação, para a S. frugiperda e S. eridania; PI 134418 e LA 462 são moderadamente resistentes a S. eridania; PI 134417 é susceptível a S. frugiperda e S. eridania; Santa Clara é altamente susceptível a S. frugiperda e S. eridania.

Efeito da alimentação de Schizaphis graminum com genótipos de sorgo no desenvolvimento do predador Cycloneda sanguinea

O pulgão Schizaphis graminum (Rondani) (Hemiptera: Aphididae) é uma das principais pragas do sorgo. O objetivo deste trabalho foi determinar o efeito da alimentação deste inseto com genótipos de sorgo resistentes e suscetíveis no desenvolvimento do predador Cycloneda sanguinea (Linnaeus) (Coleoptera: Coccinellidae). O pulgão foi criado em folhas dos genótipos de sorgo GR 11111 e TX 430 x GR 111, resistentes, GB 3B, de resistência moderada e BR 007B, suscetível ao hemíptero. As larvas de C. sanguinea foram alimentadas diariamente, ad libitum, com ninfas e adultos de S. graminum. A duração das fases de desenvolvimento e a sobrevivência de C. sanguinea não foram influenciadas pelo genótipo. Apenas a duração do quarto ínstar, da fase larval e do período de larva a adulto foram afetadas. O peso de larvas de segundo ínstar de C. sanguinea não foi afetado pelo genótipo. Nos demais ínstares e fase adulta, esse parâmetro foi diferenciado conforme o genótipo de sorgo utilizado como hospedeiro ao pulgão. Os genótipos resistentes GR 11111 e TX 430 x GR 111 não afetam adversamente o desenvolvimento e a fecundidade de C. sanguinea, durante uma geração. Estes genótipos demonstram compatibilidade com o predador e viabilizam o manejo de S. graminum na cultura do sorgo.

Adult attractiveness and oviposition preference of Bemisia tabaci (Genn.) (Homoptera: Aleyrodidae) B-biotype in cotton genotypes

Considerada importante praga do algodoeiro, a mosca-branca Bemisia tabaci biótipo B pode através de sua alimentação, diminuir o vigor das plantas, trasmitir vírus e prejudicar a qualidade da fibra. Visando avaliar a resistência de genótipos de algodoeiro, Gossypium hirsutum (L.), à mosca-branca Bemisia tabaci biótipo B, realizaram-se testes de atratividade e não-preferência para oviposição, com e sem chance de escolha, em telado, a temperatura ambiente. Verificou-se baixa atratividade das plantas dos genótipos Fabrika, CNPA Ita 90, Makina, Coodetec 407 e IAC 01-639 CPA 02-24 a adultos dessa mosca-branca, o que pode representar um componente de resistência destes materiais genéticos ao inseto. Os genótipos BRS Aroeira, Coodetec 406, Fabrika e Coodetec 401 apresentaram resistência do tipo não-preferência para oviposição, nos testes com e sem chance de escolha. Os números de tricomas e de glândulas de gossipol por cm² não foram adequados para se avaliar a não-preferência para oviposição de adultos da mosca-branca em genótipos de algodoeiro.