New insights into plants co‐existence in species‐rich communities: The pollination interaction perspective

Questions

In animal‐mediated pollination, pollinators can be regarded as a limiting resource for which entomophilous plant species might interact to assure pollination, an event pivotal for their reproduction and population maintenance. At community level, spatially aggregated co‐flowering species can thus be expected to exhibit suitable suites of traits to avoid competition and ensure pollination. We explored the problem by answering the following questions: (1) are co‐flowering species specialized on different guilds of pollinators; (2) do co‐flowering pollinator‐sharing species segregate spatially; and (3) do co‐flowering pollinator‐sharing species that diverge in anther position spatially aggregate more than those that converge in anther position?

Study Site

Euganean Hills, NE Italy.

Methods

Plant composition, flowering phenology and interactions between each entomophilous plant species and pollinating insects were monitored every 15 days in 40 permanent plots placed in an area of 16 ha. We quantified the degree of flowering synchrony, pollinator‐sharing and spatial aggregation between each pair of entomophilous species. We then tested the relationship between the degree of co‐flowering, pollinator‐sharing and spatial aggregation, and between spatial aggregation and anther position.

Results

Entomophilous species converged, at least partially in flowering time, and the phenological synchronization of flowering was significantly associated with the sharing of pollinator guilds. Co‐flowering pollinator‐sharing species segregated spatially. Furthermore, co‐flowering pollinator‐sharing species that diverged in anther position aggregated more than those that converged in anther position.

Conclusions

Reproductive traits that facilitate the co‐existence of co‐flowering species include specialization on different pollinator guilds and a phenological displacement of the flowering time. Furthermore, in circumstances of increased competition due to phenological synchronization, pollinator‐sharing and spatial aggregation, the chance of effective pollination might depend on differences in anther position, resulting in a divergent pollen placement on pollinator bodies. One of the most interesting results we obtained is that the presence of one mechanism does not preclude the operation of others, and each plant species can simultaneously exhibit different strategies. Although more studies are needed, our results can provide additional information about plant–plant interactions and provide new insights into mechanisms allowing the co‐existence of a high number of plant species in local communities.

     

An in vitro approach to the study of target organ toxicity of drugs and chemicals

Summary A major goal of our laboratory has been the development of primary culture systems that retain differentiated fucntions and responses characteristic of intact tissues in vivo. Specifically, we have developed cellular models of primary cultures of rat heart, liver, and kidney cells to explore the mechanisms by which drugs or chemicals may be toxic to key organs of the body and to develop new techniques by which xenobiotics may be evaluated or identified as potential toxicants to living systems. The purpose of this paper is to describe our rationale and approach to the study of target organ toxicology with in vitro cellular systems.     

Injury to primary cultures of rat heart endothelial cells by hypoxia and glucose deprivation

Summary Primary cultures of rat heart endothelial cells were subjected to simulated conditions of ischemia: hypoxia and glucose deprivation for 4 and 24 hr. Cellular injury was evaluated by measuring changes in viability, total protein, cellular morphology, and leakage of cytoplasmic enzymes from the cells into the culture medium. Deprivation of oxygen and glucose for 4 or 24 hr did not lethally injure the cells as noted by no change in cell viability, morphology, and total protein when compared to controls. However, reversible or nonlethal cellular injury was produced as reflected by a significant release of lactate dehydro-genase (LDH) from the cells into the medium after treatment with hypoxia and glucose deprivation for 4 or 24 hr. When the cultures were deprived of glucose, but were oxygenated, cellular injury was not evident after 24 hr. Deprivation of oxygen but not glucose resulted in significant loss of LDH after 4 or 24 hr. When the cultures were allowed to recover after oxygen and glucose deprivation in complete medium containing 1000 mg glucose per l and a normal atmosphere of 20% O₂, they had levels of LDH leakage comparable to those of control cultures. This study was supported by Research Grant HL 18647 from the National Heart, Lung, and Blood Institute and by a National Chicano Council on Higher Education Post-Doctoral Fellowship awarded to D. Acosta from the Ford Foundation. Additional support was provided to D. Acosta by a Faculty Research Assignment Award from the University of Texas Research Institute.     

Ischemic myocardial injury in cultured heart cells: Leakage of cytoplasmic enzymes from injured cells

Summary An in vitro model of myocardial ischemia has been established with primary monolayer cultures of postnatal rat myocardial cells. Ischemic conditions were simulated in vitro by subjecting the myocardial cell cultures to various levels of oxygen and glucose deprivation. The experimental protocol consisted of treatment with 20% or 0% O₂ and 1000, 500 or 0 mg glucose per 1 of medium for 4 or 24 hr. Control cultures were treated with 20% O₂ and 1000 mg glucose. After the ischemic treatments, cultures of beating muscle (M) cells were evaluated for signs of injury, i.e. leakage of cytoplasmic enzymes into the culture medium. Differences were found in leakage of lactate dehydrogenase (LDH) and creatine phosphokinase (CPK) from the cultures that were exposed to partial ischemia of glucose deprivation and from those cultures that were exposed to total ischemia of oxygen and glucose deprivation. Glucose deprivation alone resulted in a slight-to-moderate loss of LDH and CPK from the cells, whereas total ischemia resulted in a significant release of the two cytoplasmic enzymes. When the cultures were allowed to recover after ischemic treatment in complete medium (1000 mg glucose) and a normal atmosphere of 20% O₂, they had levels of LDH leakage comparable to those of control cultures. Cell viability and total protein content of the ischemic cultures did not differ significantly from controls. This study was supported by Research Grant HL 18647 from the National Heart, Lung, and Blood Institute.     

Molecular population genetics of the alcohol dehydrogenase locus in the Hawaiian drosophilid D. mimica

Sequence variation among 10 alleles of the alcohol dehydrogenase (Adh) gene of the Hawaiian drosophilid D. mimica was analyzed with reference to the evolutionary history of the Hawaiian subgroup as well as to levels and patterns of polymorphism of the Adh gene in continental drosophilid species. The Adh gene of D. mimica is less polymorphic than that of other drosophilid species, and no replacement substitutions were found. Statistical analyses of the Adh alleles suggested the action of balancing selection and revealed significant linkage disequilibrium among three of the variable sites. The effective population size was estimated to be only slightly smaller than that of continental species and, surprisingly, on the same order of magnitude as the actual size.      

Temperature-vegetation structure interaction: the effect on the activity of the ant Messor barbarus (L.)

Several canopy types in an evergreen oak forest with open pastures have been used to examine climatic features of this system related to temperature. Circadian variation in temperature seems to be related to structural characteristics of the vegetation, with an apparent influence of the plant micro- and meso-environments analyzed. The activity of a population of granivorous ants (Messor barbarus (L.)), studied in the same area, showed a clear dependence on ambient temperature, but, although the colonies were always inside one of the vegetation types, there was no significant relationship between the activity of ants and grasscover. This, together with other observations, indicates that the possible controlling effect of the vegetation is limited by the tolerance of ants (when faced with adverse conditions) during each daily foraging period.     

A kinetic study on the suicide inactivation of peroxidase by hydrogen peroxide

In the absence of reductant substrates, and with excess H2O2, peroxidase (donor: hydrogen-peroxide oxidoreductase, EC 1.11.1.7) shows the kinetic behaviour of a suicide inactivation, H2O2 being the suicide substrate. From the complex (compound I-H2O2), a competition is established between two catalytic pathways (the catalase pathway and the compound III-forming pathway), and the suicide inactivation pathway (formation of inactive enzyme). A kinetic analysis of this system allows us to obtain a value for the inactivation constant, ki = (3.92 +/- 0.06) x 10(-3) x s-1. Two partition ratios (r), defined as the number of turnovers given by one mol of enzyme before its inactivation, can be calculated: (a) one for the catalase pathway, rc = 449 +/- 47; (b) the other for the compound III-forming pathway, rCoIII = 2.00 +/- 0.07. Thus, the catalase activity of the enzyme and, also, the protective role of compound III against an H2O2-dependent peroxidase inactivation are both shown to be important.     

Distribution of indole-3-acetic acid in relation to the growth of etiolated Lupinus albus hypocotyls

The free indole-3-acetic acid (IAA) in methanolic extracts of etiolated hypocotyls of lupin ( Lupinus albus L., from Bari, Italy) was determined by fluorimetry. The distribution of IAA along the hypocotyls was parallel to the growth, but when growth ceased oscillations occurred in the auxin level. These oscillations could be related to processes of differentiation mediated by IAA. The oscillations did not obey any impulses from the apex, since the application of [1-¹⁴C]-IAA to decapitated plants gives a distribution of radioactivity which also presents an undulatory pattern. Our results support the hypothesis that morphogenesis can be regulated by information transmitted by the translocation of waves of auxin.     

Functional features and ontogenetic changes in reproductive allocation and partitioning strategies of plant modules

In the concept of modularity, plant modules are considered as iterative units and their changes are analyzed in terms of number or size. This paper, however, analyses changes with respect to the reproductive functional performance of modules and individual plant age. Patterns of resource allocation and partitioning in reproductive modules (fruits) are compared between two different age groups of a bushy perennial plant, Cistus ladanifer.Although modules do not differ in their allocation strategies (young plant modules produce the same seed and packing/protective structure biomass as old plant modules), their partitioning strategies change with plant age: young plant modules produce a larger number of lighter seeds than old plants. These differences have a direct consequence on the plant pre-dispersal fitness, which is not counteracted by insect predation on reproductive modules. These results are empirical evidence of a differentiation in the performance of reproductive modules with the ontogenetic development of this plant species. We think that the consideration of such kind of changes in module features is essential in the analysis of the iterative construction of plants.