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The expansion of sugarcane plantations in Brazil and the discarding of vinasse into the sugarcane field have been speculated to contribute to the growing population of the billbug Sphenophorus levis . This beetle attacks the root system and forms galleries in rhizomes, causing damage or even the death of host plants. It has been suspected that vinasse, a residue from ethanol production, can release volatiles that are attractive to the insect; however, no study has addressed this hypothesis so far. The aim of this study was to investigate the attractiveness of vinasse to S. levis adults and identify the volatile compounds released by this substance using gas chromatography–mass spectrometry (GC–MS). We found that vinasse was more attractive to S. levis than sugarcane stems, molasses and wastewater under laboratory conditions, but not than cane stems at field conditions. Our GC–MS analysis revealed the presence of primary alcohols, terpenes and organic carboxylic acids in vinasse. When a mixture of the commercial synthetic compounds identified in the chemical analysis was tested in the laboratory, a strong attraction of the insects to the mixture was observed. Our results help to explain how vinasse can contribute to the infestations of S. levis in sugarcane fields and shed new light on the development of strategies to control this pest using chemical attractants.

A number of soil-borne microorganisms, such as mycorrhizal fungi and rhizobacteria, establish mutualistic interactions with plants, which can indirectly affect other organisms. Knowledge of the plant-mediated effects of mutualistic microorganisms is limited to aboveground insects, whereas there is little understanding of what role beneficial soil bacteria may play in plant defense against root herbivory. Here, we establish that colonization by the beneficial rhizobacterium Azospirillum brasilense affects the host selection and performance of the insect Diabrotica speciosa. Root larvae preferentially orient toward the roots of non-inoculated plants versus inoculated roots and gain less weight when feeding on inoculated plants. As inoculation by A. brasilense induces higher emissions of (E)-β-caryophyllene compared with non-inoculated plants, it is plausible that the non-preference of D. speciosa for inoculated plants is related to this sesquiterpene, which is well known to mediate belowground insect-plant interactions. To the best of our knowledge, this is the first study showing that a beneficial rhizobacterium inoculant indirectly alters belowground plant-insect interactions. The role of A. brasilense as part of an integrative pest management (IPM) program for the protection of corn against the South American corn rootworm, D. speciosa, is considered.

The development of insects is strongly influenced by their resident microorganisms. Symbionts play key roles in insect nutrition, reproduction, and defense. Bacteria are important partners due to the wide diversity of their biochemical pathways that aid in the host development. We present evidence that the foam produced by nymphs of the spittlebug Mahanarva fimbriolata harbors a diversity of bacteria, including some that were previously reported as defensive symbionts of insects. Analysis of the microbiomes in the nymph gut and the soil close to the foam showed that the microorganisms in the foam were more closely related to those in the gut than in the soil, suggesting that the bacteria are actively introduced into the foam by the insect. Proteobacteria, Actinobacteria, and Acidobacteria were the predominant groups found in the foam. Since members of Actinobacteria have been found to protect different species of insects by producing secondary metabolites with antibiotic properties, we speculate that the froth produced by M. fimbriolata may aid in defending the nymphs against entomopathogenic microorganisms.

Insects have evolved multiple mechanisms to adapt to variations in environmental temperatures, including postural control of solar input, variations in diurnal activity, external morphological structures and selecting/generating microhabitats. Foam produced by Mahanarva fimbriolata nymphs (also known as root spittlebugs) was found to aid in creating a constant thermal microhabitat despite environmental temperature fluctuations. The temperature within the foam was found to be similar to that of soil during the day and remained constant despite fluctuating external temperatures. In chemically analysing the composition of the foam, palmitic and stearic acids, carbohydrates and proteins were detected. These substances have previously been shown to act as a surfactant to stabilize and modulate foams. Since the immature ancestor of the spittlebug developed below ground, it is speculated that the foam may function as an ‘extension’ of the soil and, thus, may have enabled the spittlebug to emerge from the soil and adopt an epigean lifestyle.

During field screening trials conducted in Brazil in 2015, adults of both sexes of the cerambycid beetles Cotyclytus curvatus (Germar) and Megacyllene acuta (Germar) (subfamily Cerambycinae, tribe Clytini) were significantly attracted to racemic 3-hydroxyhexan-2-one and racemic 2-methylbutan-1-ol, chemicals which previously have been identified as male-produced aggregation-sex pheromones of a number of cerambycid species endemic to other continents. Subsequent analyses of samples of beetle-produced volatiles revealed that males of C. curvatus sex-specifically produce only (R)-3-hydroxyhexan-2-one, whereas males of M. acuta produce the same compound along with lesser amounts of (2S,3S)-2,3-hexanediol and (S)-2-methylbutan-1-ol. Follow-up field trials showed that both sexes of both species were attracted to synthetic reconstructions of their respective pheromones, confirming that males produce aggregation-sex pheromones. The minor pheromone components of M. acuta, (S)-2-methylbutan-1-ol and (2S,3S)-2,3-hexanediol, synergized attraction of that species, but antagonized attraction of C. curvatus to (R)-3-hydroxyhexan-2-one. Beetles of other cerambycine species also were attracted in significant numbers, including Chrysoprasis linearis Bates, Cotyclytus dorsalis (Laporte & Gory), and Megacyllene falsa (Chevrolat). Our results provide further evidence that 3-hydroxyhexan-2-one is a major component of attractant pheromones of numerous cerambycine species world-wide. Our results also highlight our increasing understanding of the crucial role of minor pheromone components in imparting species specificity to cerambycid pheromone blends, as is known to occur in numerous species in other insect families.

A number of soil-borne microorganisms, such as mycorrhizal fungi and rhizobacteria, establish mutualistic interactions with plants, which can indirectly affect other organisms. Knowledge of the plant-mediated effects of mutualistic microorganisms is limited to aboveground insects, whereas there is little understanding of what role beneficial soil bacteria may play in plant defense against root herbivory. Here, we establish that colonization by the beneficial rhizobacterium Azospirillum brasilense affects the host selection and performance of the insect Diabrotica speciosa. Root larvae preferentially orient toward the roots of non-inoculated plants versus inoculated roots and gain less weight when feeding on inoculated plants. As inoculation by A. brasilense induces higher emissions of (E)- beta -caryophyllene compared with non-inoculated plants, it is plausible that the non-preference of D. speciosa for inoculated plants is related to this sesquiterpene, which is well known to mediate belowground insect-plant interactions. To the best of our knowledge, this is the first study showing that a beneficial rhizobacterium inoculant indirectly alters belowground plant-insect interactions. The role of A. brasilense as part of an integrative pest management (IPM) program for the protection of corn against the South American corn rootworm, D. speciosa, is considered.