Three–way interactions between Acacia, large mammalian herbivores and bruchid beetles - a review

Large mammalian herbivores are both predators and dispersers of Acacia seeds. While some of the seeds are destroyed during passage through the herbivore's digestive tract, others are defecated unharmed. Ingestion by large herbivores facilitates germination by scarification of the seed coat. The extent of the influence of herbivores on seed dispersal and germination depends on seed retention time and tooth size, which are both positively correlated with body size. Infestation by bruchid beetles (Bruchidae) reduces Acacia germination. Herbivores may reduce bruchid infestation in several ways. Larvae in recently infested seeds are killed by stomach acids penetrating the seed through the larval entry hole. Seeds that are partly excavated by burrowing larvae in more advanced stages may be crushed by the herbivore's teeth. Lastly, but probably most crucially, herbivores simply remove seeds from the natal tree prior to infestation or at least prior to reinfestation. The timing and magnitude of herbivory is crucial for both the reduction of bruchid infestation and Acacia seedling establishment. Although it is widely agreed that a three–way interaction exists between bruchid beetles, Acacia trees and large mammalian herbivores, it is also apparent that the relationship is highly complex and is not yet completely understood.     

The relationship between ions, vigour and salinity tolerance of Acacia seeds

Seeds of A. bivenosa DC. Prod., A. coriacea DC., A. elata A. Cunn. Ex Benth., A. farnesiana (L.) Willd., A. nilotica Willd. ex Del., A. salicina Lindl., A. saligna (Labill.) H. Wendle., A. senegal (L.) Willd., A. tortilis (Forsk.) Hayne and A. tumida F. Muell. ex Benth. were tested for their final germination percentage (FG) in distilled water (DW) and in 25–400 mol m^-3 NaCl, germination rate (1/t₅₀, where t₅₀ is the time to 50% of final germination in DW), Ca and K concentration (Ca_unt, K_unt), the leakage of these ions when soaked for 24 h in DW (Ca_DW, K_DW) and the leakage (additional to that in DW) when soaked in 250 mol m^-3 NaCl (Ca_NaCl, K_NaCl). Linear regression revealed significant positive relationships between salinity tolerance (I₅₀, the concentration of NaCl required to reduce final germination to 50% of the control value in DW) and FG, Rate, Ca_unt/K_unt and Ca_NaCl/K_NaCl. There was also a significant negative relationship between I₅₀ and K_unt. Multiple regression equations were developed to predict I₅₀ from the above parameters. Equations based on (1) FG and Rate, (2) Ca_unt and K_unt, and (3) any of these four factors, accounted for 61, 60 and 79%, respectively, of the variation in I₅₀. Further equations, introducing the leakage of ions into DW and NaCl and the ratios of ion concentrations monitored, improved the predictive value of the equation. The best equation, accounting for 93% of the variation was: I₅₀ = 108 + 422 Rate − 1.32 K_unt + 16.3 Ca_DW − 5.4 K_DW + 71.6 K/Ca_DW. These equations provide a screening test for salt tolerance of Acaciaseed germination based on simple and rapid laboratory chemical analyses and germination tests. This revised version was published online in June 2006 with corrections to the Cover Date.     

Lifecycle of the bruchid beetle Bruchidius uberatus and its predation of Acacia nilotica seeds in a tree savanna in Botswana

The life cycle of the bruchid beetle Bruchidius uberatus in seeds of Acacia nilotica has been investigated under field and laboratory condition. In contrast to the multivoltine populations in the Sudan, in Botswana most beetles of B. uberatus are univoltine. They emerge from the seed and pod in early spring (October to November). Only a small part of the population, which emerge in late summer (February to March) is multivoltine, as confirmed by rearing experiments. The reproductive activity of females is not stimulated by pollen, as found in feeding experiments. Minimum life-span of adult beetles varies between 4 and 40 days, but it did not differ between univoltine and multivoltine beetles. Other data on life-history are 22 days for hatching of the fist-instar larva at a temperature regime of 20/15 °C (day/night) and 15 days at a temperature regime of 32/15 °C and 3 to 11 months for development from larvae to beetle.During dry storage of seeds B. uberatus can destroy the total amount of stored seeds within a few years. In stored seed pool, host-specificity of larva is low.The life cycles of the Botswana population have been discussed in relation to that of the species in the Sudan.

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Zusammenfassung Der Lebenszyklus des Samenkäfers Bruchidius uberatus in Samen von Acacia nilotica wurde unter Feld- und Laborbedingungen untersucht. Im Gegensatz zu multivoltinen Populationen im Sudan (Peake, 1982) sind die Populationen von B. uberatus in Botswana vor allem univoltin. Adulte Käfer verlassen die Samen von A. nilotica während des Süd-Frühlings (Oktober/November). Im Spätsommer (Februar/März) geschlüpfte Käfer entstammen multivoltinen Genotypen, wie Züchtungsexperimente gezeigt haben. Die reproduktive Aktivität der Weibchen wird nicht durch die Zufuhr von Pollen oder Zucker stimuliert.Die Entwicklungsdauer vom Ei bis zum 1. Larvenstadium beträgt 15 Tage bis 52 und 3 bis 11 Monate bis zum adulten Käfer, deren Lebenserwartung unter Laboratoriumbedingungen 4 bis 40 Tage beträgt. Hierbei besteht kein Unterschied zwischen uni- und multivoltinen Käfern. Der lange Lebenszyklus in Botswana steht im Gegensatz zu dem 40-tägigen Lebenszyklus von B. uberatus im Sudan, der offensichtlich durch die Jahrhunderte andauernde Sammlung von Früchten verursacht ist.Samenvorräte, die zu Forstzwecken längere Zeit aufbewahrt werden, können innerhalb von 5 bis 8 Jahren völlig zerstört werden. Dabei ist die Wirtsspezifität gering. Die Infektion betrifft auch andere Acacia-Arten, z.B. A. burkei, A. erioloba, A. hebeclada, A. mellifera, A. robusta und A. tortilis. Durch die rasche Zerstörung der Embryoachse haben die Käfer keinen positiven Einfluss auf die Keimung dieser hartschaligen Acacia-Samen.

     

Ectomycorrhizal symbiosis enhanced the efficiency of inoculation with two Bradyrhizobium strains and Acacia holosericea growth

Abstract Two strains of Bradyrhizobium sp., Aust 13C and Aust 11C, were dually or singly inoculated with an ectomycorrhizal fungus, Pisolithus albus to assess the interactions between ectomycorrhizal symbiosis and the nodulation process in glasshouse conditions. Sequencing of strains Aust 13C and Aust 11C confirmed their previous placement in the genus Bradyrhizobium. After 4 months culture, the ectomycorrhizal symbiosis promoted plant growth and the nodulation process of both Bradyrhizobium strains, singly or dually inoculated. PCR/RFLP analysis of the nodules randomly collected in each treatment with Aust 13C and/or Aust 11C: (1) showed that all the nodules exhibited the same patterns as those of the Bradyrhizobium strains, and (2) did not detect contaminant rhizobia. When both Bradyrhizobium isolates were inoculated together, but without P. albus IR100, Aust 11C was recorded in 13% of the treated nodules compared to 87% for Aust 13C, whereas Aust 11C and Aust 13C were represented in 20 and 80% of the treated nodules, respectively, in the ectomycorrhizal treatment. Therefore Aust 13C had a high competitive ability and a great persistence in soil. The presence of the fungus did not significantly influence the frequencies of each Bradyrhizobium sp. root nodules. Although the mechanisms remain unknown, these results showed that the ectomycorrhizal and biological nitrogen-fixing symbioses were very dependent on each other. From a practical point of view, the role of ectomycorrhizal symbiosis is of great importance to N₂ fixation and, consequently, these kinds of symbiosis must be associated in any controlled inoculation.     

Relationships between the ant Brachymyrmex obscurior (Hymenoptera, Formicidae) and Acacia pennatula (Fabaceae)

Summary. In central Mexico, the ant Brachymyrmex obscurior Forel feeds on nectar produced by extrafloral nectaries of Acacia pennatula (Schlecht. & Cham.) Benth. However, no studies have determined whether the ants visitation is related to plant nectar availability and whether ants protect A. pennatula from herbivory. The objectives of this 2-yr study (2000–2001) were to assess whether seasonal changes in ant visitation coincide with extrafloral nectar productivity in A. pennatula and to determine whether ants protect the plant. At the end of the dry season (April–June) B. obscurior was the only ant species on A. pennatula and extrafloral nectar production is limited to this period. Exclusion experiments, performed at the end of the dry season showed that A. pennatula did not receive a protective benefit when visited by ants. Branches with ants and branches where ants are excluded had similar numbers of the nonmyrmecophile leafhopper Sibovia sp. which was the only herbivore observed under natural conditions.Received 24 March 2004; revised 4 September 2004; accepted 8 September 2004.     

Comparative effects of Glomus mosseae and P fertilizer on foliar mineral composition of Acacia senegal seedlings inoculated with Rhizobium

Summary A factorial experiment with two controlled factors was conducted in the greenhouse with Acacia Senegal seedlings. The substrate was a degraded sandy soil (Dior soil) poor in available P (11 ppm — Olsen). The first controlled factor was soil sterilization, with two levels: (A) sterilized soil; (B) non-sterilized soil. The second factor was fertilization, with six levels: (1) uninoculated control; (2) inoculation with Rhizobium (ORS 1007); (3) inoculation with Glomus mosseae; (4) double inoculation with ORS 1007 and G. mosseae; (5) inoculation with ORS 1007 and 30 ppm phosphorus per plant; (6) inoculation with ORS 1007 and 60 ppm phosphours. The combination of the two factors and their levels led to 12 different plant treatments (A1–A6 and B1–B6). Compared to the control B1, the B5 and B6 treatments containing phosphorus increased: nodule dry weight about 7 times ; leaf dry weight about 4 times ; total N, P and Mg 4–5 times; total K and Ca 3–4 times. The mycorrhizal inoculation had the same positive effect on plant growth and mineral composition but with lower values. Plants inoculated with Rhizobium alone gave the lowest results. The A1 treatment gave lower values than B1. Foliar mineral contents varied within a narrow range (20–30%).     

Multiple herbivores and coevolutionary interactions in an Ipomopsis hybrid swarm

Studies focusing on pairwise interactions between plants and herbivores may not give an accurate picture of the overall selective effect of herbivory, given that plants are often eaten by a diverse array of herbivore species. The outcome of such interactions may be further complicated by the effects of plant hybridization. Hybridization can lead to changes in morphological, phenological and chemical traits that could in turn alter plant–herbivore interactions. Here we present results from manipulative field experiments investigating the interactive effects of multiple herbivores and plant hybridization on the reproductive success of Ipomopsis aggregata formosissima X I. tenuituba. Results showed that ungulate herbivores alone had a net positive effect on plant relative fitness, increasing seed production approximately 2-fold. Caterpillars had no effect on plant relative fitness when acting alone, with caterpillar-attacked plants producing the same number of flowers, fruits and seeds as the uneaten controls. Caterpillars, however, significantly reduced flower production of ungulate browsed plants. Flower production in these plants, however, was still significantly greater (approximately 1.7-fold greater) than uneaten controls, likely leading to an increase in reproductive success through the paternal component of fitness given that fruit and seed production was not significantly different from that of herbivore-free controls. Although results suggest that herbivore imposed selection is pairwise, ungulates likely have a large influence on the abundance of, and hence the amount of damage caused by, caterpillar herbivores. Thus, because of the ecological interactions between ungulates and caterpillars, selection on Ipomopsis may be diffuse rather than pairwise, assuming such interactions translate into differential effects on plant fitness as herbivore densities vary. Plant hybridization had no significant effect on patterns of ungulate or caterpillar herbivory; i.e., no significant interactions were detected between herbivory and plant hybridization for any of the fitness traits measured in this study nor did plant hybridization have any significant effect on host preference. These results may be due to patterns of introgression or the lack of species-specific differences between I. aggregate formosissima and I. tenuituba. Plant hybridization per se resulted in lowered reproductive success of white colored morphs due in part to the effects of pollination. Although it appears that there would be strong directional selection favoring darker flower colors due to the lower reproductive success of the white colored morphs in the short run, the natural distribution of hybrids suggest that over the long run selection either tends to average out or there are no fitness differences among morphs in most years due to the additive fitness effects of hawkmoth and hummingbird pollinators.     

Characterization of rhizobia isolated from Albizia spp. in comparison with microsymbionts of Acacia spp. and Leucaena leucocephala grown in China

This is the first systematic study of rhizobia associated with Albizia trees. The analyses of PCR-RFLP and sequencing of 16S rRNA genes, SDS-PAGE of whole-cell proteins and clustering of phenotypic characters grouped the 31 rhizobial strains isolated from Albizia into eight putative species within the genera Bradyrhizobium, Mesorhizobium and Rhizobium. Among these eight rhizobial species, five were unique to Albizia and the remaining three were shared with Acacia and Leucaena, two legume trees coexisting with Albizia in China. These results indicated that Albizia species nodulate with a wide range of rhizobial species and had preference of microsymbionts different from Acacia and Leucaena. The definition of four novel groups, Mesorhizobium sp., Rhizobium sp. I, Rhizobium sp. II and "R. giardinii", indicates that further studies with enlarged rhizobial population are necessary to better understand the diversity and to clarify the taxonomic relationships of Albizia-associated rhizobia.     

Parallel diversification of Australian gall-thrips on Acacia

The diversification of gall-inducing Australian Kladothrips (Insecta: Thysanoptera) on Acacia has produced a pair of sister-clades, each of which includes a suite of lineages that utilize virtually the same set of 15 closely related host plant species. This pattern of parallel insect-host plant radiation may be driven by cospeciation, host-shifting to the same set of host plants, or some combination of these processes. We used molecular-phylogenetic data on the two gall-thrips clades to analyze the degree of concordance between their phylogenies, which is indicative of parallel divergence. Analyses of phylogenetic concordance indicate statistically-significant similarity between the two clades. Their topologies also fit with a hypothesis of some degree of host-plant tracking. Based on phylogenetic and taxonomic information regarding the phylogeny of the Acacia host plants in each clade, one or more species has apparently shifted to more-divergent Acacia host-plant species, and in each case these shifts have resulted in notable divergence in aspects of the phenotype including morphology, life history and behaviour. Our analyses indicate that gall-thrips on Australian Acacia have undergone parallel diversification as a result of some combination of cospeciation, highly restricted host-plant shifting, or both processes, but that the evolution of novel phenotypic diversity in this group is a function of relatively few shifts to divergent host plants. This combination of ecologically restricted and divergent radiation may represent a microcosm for the macroevolution of host plant relationships and phenotypic diversity among other phytophagous insects.     

Stomatal conductance and transpiration of the two faces of Acacia phyllodes

Summary The daily course of stomatal conductance and transpiration was monitored on each separate face of vertical phyllodes of various acacias. The selected phyllodes had a north-south orientation so that one side faced eastwards and the other westwards. The principal measurements were made on Acacia longifolia and A. melanoxylon in Portugal in late summer and autumn, and additional measurements were made on A. ligulata and A. melanoxylon in Australia. In Portugal, irrespective of soil moisture status, conductance showed on early morning maximum with a subsequent gradual decline and sometimes a subsidiary peak in the late afternoon. Maximum conductances appeared to be a function of soil moisture status, whereas the decline in conductance in the late morning and afternoon was correlated with changes in phyllode-to-air vapour pressure deficits rather than changes in phyllode water status. The relationship of transpiration to phyllode water potential did not appear to be influenced by soil moisture status, although transpiration was less in drier soils and in the afternoons, this latter factor contributing to a marked hysteresis in the relationship. The opposing faces of the phyllodes exhibited a high degree of synchrony, showing parallel stomatal opening and closing, despite their large differences in irradiance. Stomatal conductance tended to be higher on the eastern faces in the morning and lower in the afternoon. In A. longifolia the daily average of relative conductance was much the same for both faces, but in A. melanoxylon that of the eastern face was higher and was retained even when the normal orientation of the phyllodes was reversed by turning them through 180°. Synchrony must be achieved by the stomata of both sides responding to common environmental or endogenous signals which are perceived by both surfaces with equal sensitivity.     

Ophiostomatoid fungi isolated from Japanese red pine and their relationships with bark beetles

We isolated ophiostomatoid fungi from bark beetles infesting Pinus densiflora and their galleries at 24 sites in Japan. Twenty-one ophiostomatoid fungi, including species of Ophiostoma, Grosmannia, Ceratocystiopsis, Leptographium, and Pesotum, were identified. Among these, 11 species were either newly recorded in Japan or were previously undescribed species. Some of these fungal species were isolated from several bark beetles, but other species were isolated from only a particular beetle species. Thus, it is suggested that some ophiostomatoid fungi have specific relationships with particular beetle species. In addition, fungus-beetle biplots from redundancy analysis (RDA) summarizing the effects of beetle ecological characteristics suggested that the association patterns between bark beetles and the associated fungi seemed to be related to the niches occupied by the beetles.     

Symbiotic properties of sinorhizobia isolated from Acacia and Prosopis nodules in Sudan and Senegal

The host specificity, infection process and effectiveness of nodules produced by several African sinorhizobial strains on different Acacia and Prosopis species (Leguminosae, Mimosoideae) were studied. Sinorhizobium arboris strain HAMBI 1552^T, S. kostiense strains HAMBI 1489^T and HAMBI 1493, S. saheli strain HAMBI 1496 and S. terangae bv. acaciae strain ORS 1058 induced nitrogen fixing nodules on seedlings of the following African or Latin American species (marked with ^*): A. angustissima ^* , A. mellifera, A. nilotica, A. oerfota (synonym A. nubica), A. senegal, A. seyal, A. sieberiana, A. tortilis subsp. raddiana, P. chilensis ^* , P. cineraria, P. juliflora and P. pallida ^* . All strains increased plant yield significantly compared with uninoculated seedlings watered with nitrogen-free medium, but none appeared to be superior. The sinorhizobial strains were unable to effectively nodulate Sesbania rostrata (Papilionoideae).All roots had hairs, but particularly in the case of Acacia spp. they were often sparse. After inoculation root hairs were deformed and, in general, infection in Acacia spp. occurred through short root hairs and in Prosopis spp. through longer ones. After entry, the rhizobia filled infection pockets in the root hair, which later expanded into sac-like structures. When infection threads occurred, they usually started from sac-like structures. Elongation and ramification of the nodules indicated that Acacia spp. and Prosopis spp. have indeterminate nodules. A persistent apical meristem, which is the characteristic feature of the indeterminate nodule type, was much clearer in Prosopis spp. than in Acacia spp. Sinorhizobial strains formed tumour-like structures with undifferentiated cell tissue on the Australian acacia A. holosericea and ineffective, nodule-like structures on the African P. africana.     

Overcoming dormancy in seeds with ethanol and other anesthetics

Dormancy in fall panicum (Panicum dichotomiflorum Michx.) caryopses (seeds) is overcome by imbibition at 35° C in ethanol solutions. Whereas germination in the absence of ethanol depends on active phytochrome, the seeds may germinate in darkness after treatment with 0.2 to 0.5 M ethanol. Ethanol overcomes dormancy also in seeds of several other weedy grass species. Ethyl ether, chloroform, methanol, and acetone act similarly to ethanol. We suggest that this action depends on modifyng the properties of a membrane(s) in a manner related to the actions of other anesthetics.     

Stomatal conductance and transpiration of Acacia under field conditions: similarities and differences between leaves and phyllodes

Summary Leaf diffusive conductance and transpiration rates in response to situations of high evaporative demand were measured in 40 Acacia species varying widely with regard to the morphological and anatomical characters of their assimilatory organs. The measurements took place in south-eastern and central Australia, central Africa and south-western Europe and included species of all three subgenera of Acacia Mill. Soil moisture conditions and consequently the water status of the experimental plants varied between the different measuring sites, some of which were regularly watered. All the species investigated showed a similar daily pattern of diffusive conductance with a morning peak and a subsequent decrease, which was more pronounced in plants growing under water stress, indicating a decisive stomatal regulation of transpiration. A relationship between the structure of assimilatory organs and leaf diffusive conductance or transpiration rates per unit surface area could not be detected in the Australian acacias. However, there are indications that the leaves of the non-Australian species operate on higher conductances than the foliage of the Australian ones. It is suggested that the observed differences in the performance of African and Australian acacias reflect the deciduous or evergreen nature of foliage rather than structural differences. In regard to taxon-specific differentiation this might implicate an ecophysiological character which separates the evergreen species of the geographically isolated subgenus Heterophyllum from the deciduous species of the subgenera Aculeiferum and Acacia with an overlapping area of distribution.     

Nitrogen fixation in Faidherbia albida,Acacia raddiana,Acacia senegal and Acacia seyal estimated using the 15N isotope dilution technique

A pot experiment was conducted in a greenhouse using the ¹⁵N isotope dilution method and two reference plants, Parkia biglobosa and Tamarindus indica to estimate nitrogen fixed in four Acacia species: A raddiana, A. senegal, A. seyal and Faidherbia albida (synonym Acacia albida). For the reference plants, the ¹⁵N enrichments in leaves, stems and roots were similar. With the fixing plants, leaves and stems had similar ¹⁵N enrichments; they were higher than the ¹⁵N enrichment of roots. The amounts of nitrogen fixed at 5 months after planting were similar using either reference plant. Estimates of the percentage of N derived from fixation (%Ndfa) for the above ground parts, in contrast to %Ndfa in roots, were similar to those for the whole plant. However, none of the individual plant parts estimated accurately total N fixed in the whole plant, and excluding the roots resulted in at least 30% underestimation of the amounts of N fixed. Between species, differences in N₂ fixation were observed, both for %Ndfa and total N fixed. For %Ndfa, the best were A. seyal (average, 63%) and A. raddiana (average, 62%), being at least twice the %Ndfa in A. senegal and F. albida. Because of its very high N content, A. seyal was clearly the best in total N fixed, fixing 1.62 g N plant^–1 compared to an average of 0.48 g N plant^–1 for the other Acacia species. Our results show the wide variability existing between Acacia species in terms of both %Ndfa and total N fixed: A. seyal was classified as having a high N₂ fixing potential (NFP) while the other Acacia species had a low NFP.     

Seaweed-herbivore-predator interactions: host-plant specialization reduces predation on small herbivores

Summary Because feeding specialization among marine herbivores is rare, marine communities provide a simplified system for identifying factors selecting for specialization. On Australia's Great Barrier Reef, we investigated interactions among the chemically-defended seaweed Chlorodesmis fastigiata, herbivores specialized on this alga, and potential predators of these herbivores. Chlorodesmis is a low preference food for reef fishes but appears to be the only food of the crab Caphyra rotundifrons and the ascoglossan gastropods Elysia sp. and Cyerce nigricans. The crab is found only in patches of Chlorodesmis, feeds solely on the alga, and selectively shelters in it in laboratory choice experiments. Crab grazing on the red seaweed Acanthophora spicifera was stimulated when this alga was coated with increasing concentrations of the cytotoxic diterpenoid chlorodesmin, the major secondary metabolite of Chlorodesmis. Crabs did not sequester Chlorodesmis metabolites but avoided predators by sheltering in the unpalatable alga. All crabs tethered on the reef without access to Chlorodesmis patches were rapidly eaten; those with access to Chlorodesmis patches were much less susceptible to predation. The cryptic ascoglossan Elysia sp. was found exclusively in patches of Chlorodesmis and sequestered metabolites from the alga. Living Elysia were unpalatable to the common wrasse Thalassoma lunare in laboratory assays, but the crude organic extract of Elysia did not significantly deter feeding by Thalassoma. Elysia sequestered chlorodesmin, which deterred feeding by reef fishes in field assays but was ineffective against Thalassoma in laboratory assays at 5% food dry mass. Unlike Elysia, the aposematically colored ascoglossan Cyerce nigricans sequestered Chlorodesmis metabolites in relatively small amounts, but produced larger amounts of unrelated polypropionate compounds. Cyerce were never attacked by fishes and the crude organic extract of this slug strongly deterred feeding by wrasses in laboratory assays. The dorid nudibranch Gymnodoris sp. was found only in Chlorodesmis patches and appeared to be a specialized predator on Elysia; it would not prey on Cyerce. Data from this and other recent investigations demonstrate that some small marine herbivores feed selectively or exclusively on seaweeds that are chemically defended from fishes. This association reduces predation on the herbivores and suggests that escape from and deterrence of predation may be a dominant factor selecting for specialization among these herbivores.     

Phenotypic characteristics of root-nodulating bacteria isolated from Acacia spp. grown in Libya

Thirty isolates of root-nodulating bacteria obtained from Acacia cyanophylla, A. karroo, A. cyclops, A. tortilis (subsp.raddiana), Faidherbia albida and Acacia sp., grown in different regions of Libya, were studied by performing numerical analysis of 104 characteristics. Three fast- and one slow-growing reference strains from herbaceous and woody legumes were included. Five distinct clusters were formed. The fast-growing reference strains were separated from the isolates whereas the slow-growing was included in cluster 4. With the exception of one cluster, the majority of clusters were formed regardless of the host plant or site of origin. Based on plant tests, generation times, acid production and carbon utilization the isolates were diverse (fast and slow-growing isolates). Like slow-growing isolates, most of the fast-growing isolates appeared to be non-specific, nodulated many species from the same genus notably F. albida, known to nodulate only with slow-growing strains. Most clusters grew at temperatures 35 °C and 37 °C; some grew at temperatures above 40 °C. The majority of isolates grew at acid and alkaline pH and only one isolate grew below pH 4. Most isolates were able to utilize many amino acids as nitrogen sources and to reduce nitrate. Urea was hydrolysed by all clusters. Monosaccharides and polyols were used by slow and fast-growing isolates as the only carbon sources whereas assimilation of disaccharides varied: Some isolates, like slow-growing isolates, failed to utilize these carbon sources. Most isolates were unable to utilize polysaccharides. Regarding tolerance to NaCl on agar medium, the majority of isolates were unable to grow at a concentration of 2% NaCl, but some were highly resistant and there was one isolate which grew at 8% NaCl. Most isolates were resistant to heavy metals and to antibiotics.     

Self-incompatibility in Acacia retinodes: Site of pollen-tube arrest is the nucellus

In self-incompatible Acacia retinodes Schldl. var. uncifolia J.M. Black there is no inhibition of self pollen tubes before entry into the ovule, but the frequency of fertilized embryo sacs observed after self pollination is only 0.09–0.24 of that observed after outcrossing. Fluorescence- and light-microscope studies of sectioned, squashed or cleared whole ovules indicate that most self pollen tubes are arrested within the first or second layer of cells of the nucellus. The probability that nucellar arrest represents a primitive feature of self-incompatibility is discussed.     

Facilitative interactions between two lepidopteran herbivores of Asimina

Summary Insect herbivores that require young foliage for successful larval development are often restricted to a single generation during a year by the scarcity of suitable food over most of the growing season. The major specialist herbivore attacking shrubs in the genus Asimina in Florida, Eurytides marcellus, requires young foliage for successful larval development. Field manipulations were used to investigate the role of the young foliage produced by Asimina in response to defoliation by the late-season feeder Omphalocera munroei, a second specialist herbivore of Asimina in Florida, in maintaining Eurytides populations during the summer months when young foliage is otherwise scarce. Defoliation by Omphalocera proved to be the major inducer of young growth during the summer because Omphalocera defoliated Asimina shrubs so frequently and severely. When compared to young leaves produced in the absence of damage, the teaves produced by Asimina in response to defoliation were equally as suitable as food for Eurytides larvae and as acceptable as oviposition sites by Eurytides females. The availability of young foliage in an Asimina population was correlated with the size of the associated Eurytides population. The combination of regular, severe defoliation by Omphalocera and lack of a defensive response to damage by Asimina lead to a positive affect of Omphalocera on Eurytides population size, and may be central to other facilitative interactions between herbivores as well.