Expressed sequence tags from Atta laevigata and identification of candidate genes for the control of pest leaf-cutting ants.

Leafcutters are the highest evolved within Neotropical ants in the tribe Attini and model systems for studying caste formation, labor division and symbiosis with microorganisms. Some species of leafcutters are agricultural pests controlled by chemicals which affect other animals and accumulate in the environment. Aiming to provide genetic basis for the study of leafcutters and for the development of more specific and environmentally friendly methods for the control of pest leafcutters, we generated expressed sequence tag data from Atta laevigata, one of the pest ants with broad geographic distribution in South America. Results: The analysis of the expressed sequence tags allowed us to characterize 2,006 unique sequences in Atta laevigata. Sixteen of these genes had a high number of transcripts and are likely positively selected for high level of gene expression, being responsible for three basic biological functions: energy conservation through redox reactions in mitochondria; cytoskeleton and muscle structuring; regulation of gene expression and metabolism. Based on leafcutters lifestyle and reports of genes involved in key processes of other social insects, we identified 146 sequences potential targets for controlling pest leafcutters. The targets are responsible for antixenobiosis, development and longevity, immunity, resistance to pathogens, pheromone function, cell signaling, behavior, polysaccharide metabolism and arginine kynase activity. Conclusion: The generation and analysis of expressed sequence tags from Atta laevigata have provided important genetic basis for future studies on the biology of leaf-cutting ants and may contribute to the development of a more specific and environmentally friendly method for the control of agricultural pest leafcutters.

A pest is a pest is a pest? The dilemma of neotropical leaf-cutting ants: Keystone taxa of natural ecosystems

Leaf-cutting ants of the genera Acromyrmex and Atta are considered the principal polyphagous pests of the Neotropics. Although some members of these genera are of economic importance, have a broad geographic distribution, and are extremely good colonizers, others are endemic and closely interact with native ecosystems. Control is generally practiced against any colony, irrespective of its taxonomic status. Indiscriminate control coupled with habitat destruction threatens endemic species with extinction, and, through habitat simplification, favors other pest species. As nests of Atta are large, having several square meters of nest surface, the endemic taxa can be easily used as environmental indicators for natural ecosystems. Likewise, the pest species can be used to detect environmental disturbance. As these ants are keystone species and easily identified by nonspecialists, efforts should be made to integrate these into viable conservation programs.

Changes in sensitivity of the isolated guinea-pig vas deferens induced by a lyophilized Phoradendron latifolium leaf infusion

The effect of a lyophilized mistletoe infusion (LMI) was studied on isolated guinea-pig vas deferens. LMI caused a contraction which was partially blocked by phentolamine but not by atropine. LMI caused a shift to the left of the norepinephrine concentration-effect curve (CEC), an effect which appeared to be blocked by atropine and was absent in animals previously treated with reserpine and α-methyl-para-tyrosine. The increase of the norepinephrine maximal response induced by LMI was not blocked by atropine or pharmacological denervation. LMI caused a shift to the right of the acetylcholine CEC and had no effect on the acetylcholine maximal response. These results suggest that the effects seem to be due mainly to the presence of potassium ion in the LMI; however, the participation of muscarinic agonist(s) of reduced intrinsic activity or some tyramine-like substance could not be ruled out.

Cellulose degradation by Leucocoprinus gongylophorus, the fungus cultured by the leaf-cutting ant Atta sexdens rubropilosa

Leucocoprinus gongylophorus, the fungus cultured by the leaf-cutting ant Atta sexdens rubropilosa, is able to degrade efficiently cellulose, microcrystaline cellulose, carboximethylcellulose, and cellobiose. Analysis of the degradation products indicate that the fungus produce extracellular β-glucosidase, exo- and endo-glucanase. The importance of cellulose degradation to the association of fungus and ant is discussed.

Microbiological changes in the nests of leaf-cutting ants fed on sesame leaves

The number of bacteria and yeasts occurring in the organic matter inside the nests were determined both in colonies of leaf-cutting ants reared on Eucaliptus alba (control) or Sesamum indicum (experimental). Sesame leaves induce imbalance and nests usually die. In control nests the number of bacteria in newer sponge and in older sponge were similar (3.6 x 105 and 1.4 x 105 CFU/g, respectively) whereas in waste deposit the mean reached 7.3 x 107 CFU/g. The most Probable Number (MPN/g) of yeasts were 2.7 x 104, 1.3 x 105/g and 2.2 x 104 for newer sponge, older sponge and waste deposit, respectively. Using material from older sponge for comparison, the number of bacteria and yeasts reared on sesame leaves showed significant differences. The number of bacteria was 3.3 x 107 CFU/g, a value close to that found in waste deposit of normal (control) nests, whereas the number of yeasts was 6.7 x 105/g. These changes in microbial populations due to the effect of sesame leaf uptake could be a significant factor in nest imbalance and mortality when leaf-cutting ants are consuming toxic plants.

Effect of constituents of the foliar wax of Didymopanax vinosum on the foraging activity of the leaf-cutting ant Atta sexdens rubropilosa

Leaf epicuticular waxes may affect substrate selection by leaf-cutting ants, and host recognition by several phytophagous insects. The influence of the crude epicuticular wax of Didymopanax vinosum E. March. (Araliaceae), and its major constituents lupeol and free primary alcohols, on substrate selection by the leaf-cutting ant Atta sexdens rubropilosa Forel, was investigated. Two ant colonies were used in laboratory bioassays. Corn flakes (zea) were impregnated with chloroform solutions of the substances being tested and fed to the adults. One of the colonies gave results, in most of the tests, not significantly different from the controls. The other colony, however, discriminated against the crude wax, a strong deterrent effect being observed from the first of a series of eight trials. The same colony was able to discriminate against lupeol after the second trial. The fraction of primary n-alcohols (22% C28, 66% C30 and 12% C32) deterred feeding only after the fifth trial. The results demonstrate that some constituents of epicuticular waxes may deter the foraging activity of A. sexdens rubropilosa, depending, however, on the colony under observation. It is suggested that lupeol, of the foliar wax of D. vinosum, is an important deterrent to leaf-cutting ants, although with less pronounced effects than those observed in tests with the crude wax.

Inhibitory effects of jackbean (Canavalia ensiformis L.) leaf residues on germination and vigour of crops and weeds

The effects of jackbean leaf residues incorporated in the soil on germination and seedlings growth of cucumber, radish and some weeds was examined. Trials were carried out under greenhouse conditions to (a) determine the amount of incorporated residue that is inhibitory to two test plants, (b) to determine if decomposition time changes the inhibitory levels of jackbean residues on test plants and (c) to determine the amount of residue that is inhibitory to the weed species. Jackbean leaf residues incorporated in soil at concentration of 2% or higher and allowed to decompose for a period of 0 to 2 weeks before sowing, reduced the initial growth of cucumber and radish and at different concentrations, reduced germination and growth of three weed species. These results suggest the presence of allelopathic components in Jackbean leaves that could affect seed germination and seedling development.

Activity of Extracts and Fatty Acids of Canavalia ensiformis (Leguminosae) Against the Symbiotic Fungus of the Leaf-Cutting Ants Atta sexdens

The inhibitory effect of leaves extracts of Carnavalia ensiformis on the development of the symbiotic fungus of the leaf-cutting ants Atta sexdens (Forel) was evaluated. The hexane extract showed highest activity at concentration of 1000 μg/mL. Chromatographic separations of this extract have led to the isolation of a mixture of fatty acids which showed the same activity of the crude extract.

Systematic vegetative anatomy and ensiform leaf development in Xyris (Xyridaceae)

Ensiform leaf development in monocotyledons follows a broadly similar sequence in a wide range of relatively unrelated taxa, indicating a plastic developmental pattern, possibly associated with stressed environmental conditions, since Xyris species tend to grow in relatively damp but nutrient-poor environments. The bifacial leaf sheath surrounds the apex and the subadjacent primordium. A conical unifacial leaf tip 'Vorlauferspitze' is established at an early stage, followed by extension growth in the region behind it, generating a unifacial ensiform blade. Root and rhizome structure are also described in a systematic context, particularly in comparison with related taxa in Xyridaceae and other commelinoid monocotyledons, although information on these structure is relatively sparse.

Application of on-line C30 RP-HPLC-NMR for the analysis of flavonoids from leaf extract of Maytenus aquifolium

The application of on-line C30-reversed-phase high-pressure liquid chromatography-nuclear magnetic resonance spectroscopy is described for the analysis of tetraglycosylated flavonoids in aqueous and hydroalcoholic extracts of the leaves of Maytenus aquifolium (Celastraceae). Triacontyl stationary phases showed adequate separation for on-line 1H-NMR measurements at 600 MHz and allowed the characterisation of these flavonoids by detection of both aromatic and anomeric proton signals. Copyright (C) 2000 John Wiley and Sons, Ltd.

Toxicity of synthetic piperonyl compounds to leaf-cutting ants and their symbiotic fungus

The development of Leucoagaricus gongylophorus, the fungus cultured by the leaf-cutting ant Atta sexdens was inhibited in vitro by synthetic compounds containing the piperonyl group. In addition, worker ants that were fed daily on an artificial diet to which these compounds were added had a higher mortality rate than the controls. The inhibition of the fungal growth increased with the size of the carbon side chain ranging from C1 through C8 and decreasing thereafter. 1-(3,4-Methylenedioxybenzyloxy)octane (compound 5) was the most active compound and inhibited the fungal development by 80% at a concentration of 15 μg m1-1. With worker ants the toxic effects started with compound 5 and increased with the number of carbons in the side chain. Thus, for the same concentration (100 μg m1-1) the mortality rates observed after 8 days of diet ingestion were 82%, 66% and 42%, for 1-(3,4-methylenedioxybenzyloxy)decane, 1-(3,4-methylenedioxybenzyloxy)dodecane and compound 5, respectively, whereas with commercial piperonyl butoxide the mortality was 68%. The latter compound, which is known as a synergist insecticide, was as inhibitory to the symbiotic fungus as the synthetic compound 5. The possibility of controlling these insects in the future using compounds that can target simultaneously both organisms is discussed. © 2001 Society of Chemical Industry.

Behavior of Atta sexdens rubropilosa (Hymenoptera: Formicidae) workers during the preparation of the leaf substrate for symbiont fungus culture

The behavioral repertory of Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae) workers marked by size category was studied during the preparation of the leaf substrate in the laboratory. The workers were marked according to three physical castes, i.e., minima, generalist and forager. Seven types of behavioral acts were recorded for each caste, with the following frequencies: licking leaf fragments (64.6%), holding fragments on the surface of the fungus garden (16.4%), shredding the fragments (6.0%), chewing and crimping the edges of the fragments (9.0%), incorporating the fragments (2.7%), touching the surface of the fungus with their mandibles and other mouthparts after incorporation (0.3%), and depositing fecal fluid (0.1%). The minima workers were found to be more specialized in the activities related to the preparation of the leaf substrate, which represented 52% of the total number of tasks performed. The generalists performed 40.3% of these tasks, and the foragers 7.9%. Licking the substrate was the behavior most frequently recorded and performed for a longer period of time. In this way, the workers may feed and at the same time eliminate microorganisms that are harmful to the symbiont fungus. The smaller castes, minima and generalists, are those most responsible for the preparation of the leaf substrate and predominate within a colony. From a practical viewpoint, with the introduction of toxic bait containing insecticides, for example, these size categories will be those most intensely intoxicated, especially through the behavior of licking bait pellets. On the basis of the data obtained about these behaviors, we may raise the hypothesis that trophallaxis in not the major factor triggering contamination with an insecticide among the workers of a colony.

Leaf and stem anatomy of Vochysiaceae in relation to subfamilial and suprafamilial systematics

Several leaf anatomical features are potentially systematically informative within both the family Vochysiaceae and the order Myrtales, notably tracheoidal idioblasts, mucilage cells and secretory canals. Tracheoids with spiral wall thickenings are present in the mesophyll of most species of Vochysia, and also occur in several other families of Myrtales. Mucilage cells are common in the leaf epidermis in some Vochysiaceae. Secretory ducts are present in the midrib in Salvertia and Vochysia, which are apparently closely related, although Salvertia also shares some leaf anatomical characters with Qualea and Callisthene. Anatomical data do not support the segregation of Ruizterania from Qualea; leaves of R. albiflora leaves are very similar to those of Q. paraensis in venation pattern, and leaf and stem anatomy. Different venation patterns are characteristic of sections within the genus Qualea, but within the large genus Vochysia, leaf anatomy is variable even within a subsection. Amongst other Myrtales, leaf anatomy of Vochysiaceae most closely resembles that of Combretaceae and Onagraceae. © 2002 The Linnean Society of London.

Occurrence of killer yeasts in leaf-cutting ant nests

Killer activity was screened in 99 yeast strains isolated from the nests of the leaf-cutting ant Atta sexdens against 6 standard sensitive strains, as well as against each other. Among this yeast community killer activity was widespread since 77 strains (78 %) were able to kill or inhibit the growth of at least one standard strain or nest strain. Toxin production was observed in representatives of all the studied genera including Aureobasidium, Rhodotorula, Tremella and Trichosporon, whose killer activity has not yet been described.

Stresses analysis in semi-elliptic leaf spring and comparison with the SAE J788 (1982) norm

In this work are presented the values found with the experimental testing, in the semi-elliptic leaf spring, utilizing 24 strain gages, distributed in five leaves of springs; these values have been compared to the calculated values found with the application of Norm SAE J788 (1982). The results showed discrepancy between the values measured and calculated and that the Norm is not indicated to determine the actuating stress in any point of any leaf of the leaf spring, but due to its simplicity and quickness of the process it presents good precision for the pre-development of the product. Copyright © 2002 Society of Automotive Engineers, Inc.