THE EXTENT of SEED PREDATION BY BRUCHINE BEETLES (COLEOPTERA: CHRYSOMELIDAE: BRUCHINAE) IN A HETEROGENEOUS LANDSCAPE IN SOUTHEASTERN BRAZIL

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

Host preference of the bean weevil Zabrotes subfasciatus

It is largely known that the range of an insect diet is mostly determined by oviposition behavior, mainly in species with endophytic larvae such as Zabrotes subfasciatus. However, the proximate factors determining host choice and the subsequent steps leading to the expansion or reduction of the host number and occasional host shifts are largely unknown. We analyzed various factors determining host preference of Z. subfasciatus through the evaluation of: (i) oviposition preference of a wild population of Z subfasciatus on the usual host (bean) and unusual hosts (lentil, chickpea and soy), and the performance of the offspring; (ii) artificial selection for increasing preference for hosts initially less frequently chosen; (iii) comparison of oviposition behavior between two different populations (reared for similar to 30 generations in beans or chickpeas, respectively); (iv) oviposition timing on usual and unusual hosts; and (v) identification of preference hierarchies. We found that when using unusual hosts, there is no correlation between performance and preference and that the preference hierarchy changes only slightly when the population passes through several generations on the less frequently accepted host. We also found a positive response to artificial selection for increasing oviposition on the less preferred host; however, when the host-choice experiment involved two varieties of the usual host, the response was faster than when the choice involved usual and unusual hosts. Finally, beetles reared on an unusual host (chickpea) for 26 generations showed similar good fitness on both usual and unusual hosts, indicating that the use of a new host does not necessarily result in the loss of performance on the original host. Nevertheless, this population showed lower fitness on the usual host than that of the original population, suggesting an underlying partial trade-off phenomenon which may contribute to a broadening of diet of this insect species.

The comparative effectiveness of rodents and dung beetles as local seed dispersers in Mediterranean oak forests

The process of seed dispersal of many animal-dispersed plants is frequently mediated by a small set of biotic agents. However, the contribution that each of these dispersers makes to the overall recruitment may differ largely, with important ecological and management implications for the population viability and dynamics of the species implied in these interactions. In this paper, we compared the relative contribution of two local guilds of scatter-hoarding animals with contrasting metabolic requirements and foraging behaviours (rodents and dung beetles) to the overall recruitment of two Quercus species co-occurring in the forests of southern Spain. For this purpose, we considered not only the quantity of dispersed seeds but also the quality of the seed dispersal process. The suitability for recruitment of the microhabitats where the seeds were deposited was evaluated in a multi-stage demographic approach. The highest rates of seed handling and predation occurred in those microhabitats located under shrubs, mostly due to the foraging activity of rodents. However, the probability of a seed being successfully cached was higher in microhabitats located beneath a tree canopy as a result of the feeding behaviour of beetles. Rodents and beetles showed remarkable differences in their effectiveness as local acorn dispersers. Quantitatively, rodents were much more important than beetles because they dispersed the vast majority of acorns. However, they were qualitatively less effective because they consumed a high proportion of them (over 95%), and seeds were mostly dispersed under shrubs, a less suitable microhabitat for short-term recruitment of the two oak species. Our findings demonstrate that certain species of dung beetles (such as Thorectes lusitanicus), despite being quantitatively less important than rodents, can act as effective local seed dispersers of Mediterranean oak species. Changes in the abundance of beetle populations could thus have profound implications for oak recruitment and community dynamics.

Females of the weevil Zabrotes subfasciatus manipulate the size and number of eggs according to the host seed availability

In some insects, the finding of oviposition substrate triggers the uptake into oocytes of yolk proteins that are stored in the fat body during post-embryonic development. The main host of the bean weevil Zabrotes subfasciatus (Coleoptera; Chrysomelidae; Bruchinae; Amblycerini), in which larval resources are the sole source for future egg maturation, is Phaseolus vulgaris. Despite not feeding as adults, females of this species are able to lay eggs after encountering host seeds but it is not known how females react to changes in the availability of bean seeds. In the present study, the behaviour of Z. subfasciatus facing two very different environments for oviposition is investigated, as well as how this influences offspring fitness. The results obtained show that females of Z. subfasciatus react to variations in the availability of seeds belonging to the same host species by adjusting egg size and number. Females on low bean seed density lay larger and fewer eggs than those on high bean seed density, demonstrating a trade-off between these reproductive traits. Moreover, females can adjust egg size to changing levels of host availability during the first 4 days of their oviposition period. Although no difference in offspring weight is found, those from small eggs (low competition environment) result in larger adults. No response to selection on these traits after rearing beetles on the same host for 40 generations is observed. This unresponsiveness may indicate that beetle populations behave according to their reaction norm that already allows rapid adaptation to a varying amount of host-seed availability and better exploitation of the environments of this widespread stored-seed pest.

Effects of seed mixture and management on beetle assemblages of arable field margins

Beetle assemblages and their response to plant community composition and architectural structure were monitored from 2002 to 2006 within arable field margins. Field margins were sown with either tussock grass and forbs, fine grass and forbs or grass only seed mixtures. After an establishment year, field margins were managed using standard sward cuts, scarification, or graminicide application. For predatory beetles, overall density was greatest where tussock grasses were included within the seed mixtures, while the densities of phytophagous beetles were greatest where forbs were present. Unexpectedly, species rarefaction curves suggested that phytophagous beetle species richness was greatest where field margins were established using a grass only seed mixture. The structure of the beetle assemblages, i.e., the relative abundances of individual species, was largely dependent on seed mixture, although margin management also played an important role. The results suggest that field margins established using seed mixtures containing tussock grasses and forbs would be expected to provide the greatest resources for beetles, at least at local scales. However, the use of a single standardised seed mixture for margin establishment would result in a homogenisation of beetle assemblages at a regional scale. (C) 2008 Elsevier B.V. All rights reserved.

The restoration of phytophagous beetles in species-rich chalk grasslands

This study focuses on the restoration of chalk grasslands over a 6-year period and tests the efficacy of two management practices, hay spreading and soil disturbance, in promoting this process for phytophagous beetles. Restoration success for the beetles, measured as similarity to target species-rich chalk grassland, was not found to be influenced by either management practice. In contrast, restoration success for the plants did increase in response to hay spreading management. Although the presence of suitable host plants was considered to dictate the earliest point at which phytophagous beetles could successfully colonized, few beetle species colonized as soon as their host plants became established. Morphological characteristics and feeding habits of 27 phytophagous beetle species were therefore tested to identify factors that limited their colonization and persistence. The lag time between host plant establishment and colonization was greatest for flightless beetles. Beetles with foliage-feeding larvae both colonized at slower rates than seed-, stem-, or root-feeding species and persisted within the swards for shorter periods. Although the use of hay spreading may benefit plant communities during chalk grassland restoration, it did not directly benefit phytophagous beetles. Without techniques for overcoming colonization limitation for invertebrate taxa, short-term success of restoration may be limited to the plants only.

Effects of seed addition on beetle assemblages during the re-creation of species-rich lowland hay meadows

Species-rich lowland hay meadows are of conservation importance for both plants and invertebrates; however, they have declined in area across Europe as a result of conversion to other land uses and management intensification. The re-creation of these grasslands on ex-arable land provides a valuable approach to increasing the extent and conservation value of this threatened habitat. Over a 3-year period a replicated block design was used to test whether introducing seeds promoted the re-creation of both plant and phytophagous beetle assemblages typical of a target hay meadow. Seeds were harvested from local hay meadows, and applied to experimental plots in the form of either green hay or brush harvesting seeds. Green hay spreading achieved the greatest success in re-creating plant and phytophagous beetle assemblages. While re-creation success increased over time for both taxa, for the phytophagous beetles the greatest increase in re-creation success relative to the establishment year also occurred where green hay was applied. We also considered the phytophagous beetles in terms of functional traits that describe host plant specificity, larval feeding location and dispersal. Phytophagous beetle functional trait composition was most similar to the target hay meadow assemblage where some form of seed addition was used, i.e. hay spreading or brush harvested seeds. This study identified the importance of introducing target plant species as a mechanism to promote the re-creation of phytophagous beetle communities. Seed addition methods (e.g. green hay spreading) are crucial to successful hay meadow re-creation.

Bottom-up and top-down effects in a pre-dispersal seed predation system: are non-predated seeds damaged?

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

Seed dispersal and spatial distribution of Attalea geraensis (Arecaceae) in two remnants of Cerrado in Southeastern Brazil

The seed dispersal system of Attalea geraensis (Arecaceae), an acaulescent palm, was investigated during one year in two Cerrado fragments in the state of São Paulo, southeastern Brazil. A. geraensis had inflorescences and infrutescences throughout the year. Two scatter-hoarding rodents (the spiny rat, Clyomys bishopi and agoutis, Dasyptocta azarae) were identified as seed predators/dispersers, able to move seeds up to 30 in from the palms, although most of the fruits (57.5%) were dispersed less than 2 in. The removal rates were high and after 20 days, 97.2% of the fruits were removed. Fruit fate was not related to fruit mass, length and diameter. The application of Morisita's index showed a more clumped distribution of adults in the smaller fragment, probably because of the absence of agoutis. Higher seed removal by rodents in the large Cerrado remnant may decrease seed predation by beetles. (C) 2007 Elsevier Masson SAS. All rights reserved.

Seed size variation in the palm Euterpe edulis and the effects of seed predators on germination and seedling survival

Intraspecific variation in seed size is common in wild plant populations and has important consequences for the reproductive success of individual plants. Multiple, often conflicting evolutionary forces mediated by biotic as well as abiotic agents may maintain such a variation. In this paper we assessed seed size variation in a population of the threatened, commercially important palm Euterpe edulis in southeast Brazil. We investigated (i) how this variation affects the probability of attack by vertebrate and invertebrate post-dispersal seed predators, and (ii) if seed size influences the outcome of seeds damaged by beetles in terms of seed germination and early survival of seedlings. Euterpe edulis seeds varied in diameter from 8.3 to 14.1 mm. Neither insects nor rodents selected the seeds they preyed upon based on seed size. Seed germination and total, shoot and root biomasses of one-year seedlings were significantly and positively affected by seed size. Root biomass and seedling survival were negatively affected by seed damage caused by a scolytid beetle (Coccotrypes palmarum) whose adults bore into seeds to consume part of the endosperm, but do not oviposit on them. Seed size had a marginally significant effect on seedling survival. Therefore, if any advantage is accrued by E. edulis individuals producing large seeds, this is because of greater seed germination success and seedling vigor. If this is so, even a relatively narrow range of variation in seed size as observed in the E. edulis population studied may translate into differential success of individual plants. (c) 2006 Elsevier SAS. All rights reserved.

A preliminary investigation of pre-dispersal seed predation by Acanthoscelides schrankiae horn (Coleoptera: Bruchidae) in Mimosa bimucronata (DC.) Kuntze trees

This is the first record of Acanthoscelides schrankiae Horn, feeding in seeds of Mimosa bimucronata (DC.) Kuntze. We investigated the pattern of oviposition and seed exploitation by A. schrankiae, and the distribution of mature fruits and seed predation in the inflorescences. We also compared the percentage of predated seeds, the total dry weight of fruits and non-predated seeds, the percentage of aborted seeds, and the percentage of non-emergent insects, among different quadrants of the M. bimucronata canopy. To determine the occurring species, the emergence of bruchids and parasitoids was observed in the laboratory, resulting altogether, only in individuals of A. schrankiae and Horismenus sp. (Hymenoptera: Eulophidae) species, respectively. Mean number of fruits produced in the median region of inflorescence was significantly higher than in the inferior and superior regions, and the frequencies (observed and expected) of predated and non-predated seeds differed among the different regions of inflorescence. Females of A. schrankiae laid their eggs on fruits, and larvae, after emergence, perforated the exocarp to reach the seeds. Most fruits presented one to three eggs and only one bruchid larva was observed in each seed. The highest value of the rate number of eggs/fruit and the highest percentage of predated seeds were recorded in April. Dry weight of fruits (total) and seeds (non-predated), proportions of predated seeds, seed abortions, and non-emergent seed predators, were evenly distributed in the canopy.

Partitioning the relative contribution of one-phase and two-phase seed dispersal when evaluating seed dispersal effectiveness

Seed dispersal effectiveness (SDE) is a conceptual framework that aims at quantifying the contribution of seed dispersal vectors to plant fitness. While it is well recognized that diplochorous dispersal systems, characterized by two successive dispersal steps performed by two different vectors (Phase I=primary seed dispersal and Phase II=secondary seed dispersal) which are common in temperate and tropical regions, little attention has been given to distinguishing the relative contribution of one-phase and two-phase dispersal to overall SDE. This conceptual gap probably results from the lack of a clear methodology to include Phase II dispersal into the calculation of SDE and to quantify its relative contribution. We propose a method to evaluate the relative contribution of one-phase and two-phase dispersal to SDE and determine whether two seed dispersers are better than one. To do so, we used the SDE landscape and an extension of the SDE landscape, the Phase II effect landscape, which measures the direction and magnitude of the Phase II dispersal effect on overall SDE. We used simulated and empirical data from a diplochorous dispersal system in the Peruvian Amazon to illustrate this new approach. Our approach provides the relative contribution of one-phase SDE (SDE1) and two-phase SDE (SDE2) to overall SDE and quantifies how much SDE changes with the addition of Phase II dispersal. Considering that the seed dispersal process is context dependent so that Phase II depends on Phase I, we predict the possible range of variation of SDE according to the variation of the probability of Phase II dispersal. In our specific study system composed of two primate species as primary dispersal vectors and different species of dung beetles as secondary dispersal vectors, the relative contribution of SDE1 and SDE2 to overall SDE varied between plant species. We discuss the context dependency of the Phase II dispersal and the potential applications of our approach. This extension to the conceptual framework of SDE enables quantitative evaluation of the effect of Phase II dispersal on plant fitness and can be easily adapted to other biotic and/or abiotic diplochorous dispersal systems.

Gut content analysis to distinguish between seed feeding and mycophagy of a biphyllid beetle larva found on Acacia melanoxylon

In a search for potential biocontrol agents for Acacia melanoxylon R. Br. (Mimosaceae), larvae of the beetle Diplocoelus dilataticollis Lea (Coleoptera; Biphyllidae) were found within damaged seeds of A. melanoxylon. The gut contents of larvae and adults were examined to determine whether their diet included seeds, in apparent contradiction to the known mycophagous diet of members of this family of beetles. Calcofluor M2R White, a plant cell-wall staining optical brightener was used to differentiate between plant cell fragments and fungal tissue in the gut content smears. Gut contents of adults of a known seed predator of A. melanoxylon, a weevil of the genus Melanterius, were examined in the same way to provide a benchmark. The gut contents of D. dilataticollis differed from those of Melanterius sp. Fungal structures and microbes were found in the gut of D. dilataticollis, in contrast to plant cell fragments found in the gut of the weevil and from scrapes made directly from seeds. We conclude that larvae of D. dilataticollis feed primarily on fungi associated with damaged seed and therefore may not be the proximate cause of seed damage.

The Cratylia Mollis Seed Lectin Induces Membrane Permeability Transition In Isolated Rat Liver Mitochondria And A Cyclosporine A-insensitive Permeability Transition In Trypanosoma Cruzi Mitochondria.

Previous results provided evidence that Cratylia mollis seed lectin (Cramoll 1,4) promotes Trypanosoma cruzi epimastigotes death by necrosis via a mechanism involving plasma membrane permeabilization to Ca(2+) and mitochondrial dysfunction due to matrix Ca(2+) overload. In order to investigate the mechanism of Ca(2+) -induced mitochondrial impairment, experiments were performed analyzing the effects of this lectin on T. cruzi mitochondrial fraction and in isolated rat liver mitochondria (RLM), as a control. Confocal microscopy of T. cruzi whole cell revealed that Cramoll 1,4 binding to the plasma membrane glycoconjugates is followed by its internalization and binding to the mitochondrion. Electrical membrane potential (∆Ψm ) of T. cruzi mitochondrial fraction suspended in a reaction medium containing 10 μM Ca(2+) was significantly decreased by 50 μg/ml Cramoll 1,4 via a mechanism insensitive to cyclosporine A (CsA, membrane permeability transition (MPT) inhibitor), but sensitive to catalase or 125 mM glucose. In RLM suspended in a medium containing 10 μM Ca(2+) this lectin, at 50 μg/ml, induced increase in the rate of hydrogen peroxide release, mitochondrial swelling, and ∆Ψm disruption. All these mitochondrial alterations were sensitive to CsA, catalase, and EGTA. These results indicate that Cramoll 1, 4 leads to inner mitochondrial membrane permeabilization through Ca(2+) dependent mechanisms in both mitochondria. The sensitivity to CsA in RLM characterizes this lectin as a MPT inducer and the lack of CsA effect identifies a CsA-insensitive MPT in T. cruzi mitochondria.

Seed dispersal of Solanum thomasiifolium Sendtner (Solanaceae) in the Linhares Forest, Espírito Santo state, Brazil

The aim of this study was to analyse seed dispersal and establishment of Solanum thomasiifolium in an area of nativo vegetation in Espirito Santo state on the southeastern Brazilian coast. Ten species of birds, the crab-eating fox (Cerdocyon thous), and one species of lizard (Tropidurus torquatus) fed on S. thomasiifolium fruits and dispersed viable seeds in their faeces. The proportional contribution of each of these groups to seed dispersal was 77% (birds), 19% (crab-eating fox) and 4% (lizards). Ants also contributed to seed dispersal. More seeds were deposited in vegetation islands than in the surrounding open areas. Germination rates of seeds collected directly from fruit (control), bird droppings, the faeces of crab-eating foxes and lizards were, respectively, 64, 64, 53, and 80 %. Differences among these rates were all significant, except between birds and control. Lizards were important as seed carriers between nearby islands and they expelled a higher proportion of viable seeds. Birds and the crab-eating foxes did not enhance seed germination, but promoted seed dispersal over a wider area. Plant architecture, fruit productivity, fruit characteristics and the diversity of frugivores are important for the success of S. thomasiifolium in habitat colonization.