Evolutionarily conserved plant genes responsive to root-knot nematodes identified by comparative genomics.

Root-knot nematodes (RKNs, genus Meloidogyne) affect a large number of crops causing severe yield losses worldwide, more specifically in tropical and sub-tropical regions. Several plant species display high resistance levels to Meloidogyne, but a general view of the plant immune molecular responses underlying resistance to RKNs is still lacking. Combining comparative genomics with differential gene expression analysis may allow the identification of widely conserved plant genes involved in RKN resistance. To identify genes that are evolutionary conserved across plant species, we used OrthoFinder to compared the predicted proteome of 22 plant species, including important crops, spanning 214 Myr of plant evolution. Overall, we identified 35,238 protein orthogroups, of which 6,132 were evolutionarily conserved and universal to all the 22 plant species (PLAnts Common Orthogroups-PLACO). To identify host genes responsive to RKN infection, we analyzed the RNA-seq transcriptome data from RKN-resistant genotypes of a peanut wild relative (Arachis stenosperma), coffee (Coffea arabica L.), soybean (Glycine max L.), and African rice (Oryza glaberrima Steud.) challenged by Meloidogyne spp. using EdgeR and DESeq tools, and we found 2,597 (O. glaberrima), 743 (C. arabica), 665 (A. stenosperma), and 653 (G. max) differentially expressed genes (DEGs) during the resistance response to the nematode. DEGs' classification into the previously characterized 35,238 protein orthogroups allowed identifying 17 orthogroups containing at least one DEG of each resistant Arachis, coffee, soybean, and rice genotype analyzed. Orthogroups contain 364 DEGs related to signaling, secondary metabolite production, cell wall-related functions, peptide transport, transcription regulation, and plant defense, thus revealing evolutionarily conserved RKN-responsive genes. Interestingly, the 17 DEGs-containing orthogroups (belonging to the PLACO) were also universal to the 22 plant species studied, suggesting that these core genes may be involved in ancestrally conserved immune responses triggered by RKN infection. The comparative genomic approach that we used here represents a promising predictive tool for the identification of other core plant defense-related genes of broad interest that are involved in different plant-pathogen interactions.

Observing the devastating coffee berry borer (Hypothenemus hampei) inside the coffee berry using micro-computed tomography.

The coffee berry borer is the most devastating insect pest of coffee throughout the world. The insect spends most of its life cycle inside the coffee berry, which makes it quite difficult to observe its behaviour. Micro-computed tomography (micro-CT) was used to observe all developmental stages of the coffee berry borer inside coffee berries (Coffea canephora). An interesting oviposition pattern involving a sequential placement of eggs starting in the periphery of the seed and moving inwards was observed. Micro-CT should be useful in elucidating unknown life history aspects of other seed-feeding bark beetles as well as of bark and ambrosia beetles in general.

Responses of aerial insectivorous bats to local and landscape-level features of coffee agroforestry systems in Western Ghats, India.

Shade coffee has shown great promise in providing crucial habitats for biodiversity outside formal protected areas. Insectivorous bats have been understudied in coffee, although they may provide pest control services. We investigated the influence of local and landscape-level features of coffee farms on aerial insectivorous bats in Chikmagalur district in the Western Ghats biodiversity hotspot, India. Bats were monitored in 20 farm sites using ultrasound detectors, and the response of bat species richness and activity to changes in tree density, proportion of built-up area in the neighborhood, and distance of farm from forest areas quantified. We examined if models built to explain the species richness and activity could also predict them in nine additional sites. We detected nine phonic types/species in the study area. The quantified predictors had no effect on assemblage-level species richness and activity of bats. Responses of edge-space and cluttered-space forager guilds mirrored those of the overall assemblage, but some species vulnerable to forest conversion like Rhinolophus beddomei were detected rarely. Best models explained up to 20% and 15% variation in assemblage-level species richness and activity respectively, and were poor predictors of both response variables. We conclude that coffee farms in our study area offer an important commuting space for insectivorous bats across a gradient of shade management. Further research should include species-specific responses to management decisions for at-risk species and quantification of ecosystem services like natural pest control to inform biodiversity conservation initiatives in the Western Ghats coffee landscapes.

Scale insects (Hemiptera: Coccoidea) associated with arabica coffee and geographical distribution in the neotropical region.

Coffee is one of the most important Brazilian agricultural commodities exported, and Minas Gerais and Espírito Santo States are the main coffee producers. Scale insects are important coffee pests, and 73 species of Cerococcidae (3), Coccidae (18), Diaspididae (6), Eriococcidae (1), Ortheziidae (3), Pseudococcidae (21), Putoidae (2) and Rhizoecidae (19) have been associated with roots, branches, leaves, flowers and fruits of Arabica coffee in the Neotropics. Eight species were found associated with Arabica coffee in Minas Gerais and Espírito Santo States in this study, and Coccidae was the most frequent family. Coccus alpinus, Cc. celatus, Cc. lizeri, Cc. viridis, and Saissetia coffeae (Coccidae) were found in both states; Alecanochiton marquesi, Pseudaonidia trilobitiformis (Diaspididae), and Dysmicoccus texensis (Pseudococcidae) were only found in Minas Gerais. Alecanochiton marquesi and P. trilobitiformis are first reported in Minas Gerais, and Cc. alpinus in Espírito Santo, on Arabica coffee. All scale insect species were associated with coffee leaves and branches, except D. texensis, associated with coffee roots. Fourty seven scale insect species have been found occurring in Brazilian Arabica coffee, and in Espírito Santo (28) and Minas Gerais (23). Widespread and geographical distribution of each species found are discussed.

Scale insects (Hemiptera: Coccoidea) associated with arabica coffee and geographical distribution in the neotropical region

ABSTRACT Coffee is one of the most important Brazilian agricultural commodities exported, and Minas Gerais and Espírito Santo States are the main coffee producers. Scale insects are important coffee pests, and 73 species of Cerococcidae (3), Coccidae (18), Diaspididae (6), Eriococcidae (1), Ortheziidae (3), Pseudococcidae (21), Putoidae (2) and Rhizoecidae (19) have been associated with roots, branches, leaves, flowers and fruits of Arabica coffee in the Neotropics. Eight species were found associated with Arabica coffee in Minas Gerais and Espírito Santo States in this study, and Coccidae was the most frequent family. Coccus alpinus, Cc. celatus, Cc. lizeri, Cc. viridis, and Saissetia coffeae (Coccidae) were found in both states; Alecanochiton marquesi, Pseudaonidia trilobitiformis (Diaspididae), and Dysmicoccus texensis (Pseudococcidae) were only found in Minas Gerais. Alecanochiton marquesi and P. trilobitiformis are first reported in Minas Gerais, and Cc. alpinus in Espírito Santo, on Arabica coffee. All scale insect species were associated with coffee leaves and branches, except D. texensis, associated with coffee roots. Fourty seven scale insect species have been found occurring in Brazilian Arabica coffee, and in Espírito Santo (28) and Minas Gerais (23). Widespread and geographical distribution of each species found are discussed.

Local and Landscape Constraints on Coffee Leafhopper (Hemiptera: Cicadellidae) Diversity.

The intensification of agriculture drives many ecological and environmental consequences including impacts on crop pest populations and communities. These changes are manifested at multiple scales including small-scale management practices and changes to the composition of land-use types in the surrounding landscape. In this study, we sought to examine the influence of local and landscape-scale agricultural factors on a leafhopper herbivore community in Mexican coffee plantations. We sampled leafhopper (Hemiptera: Cicadellidae) diversity in 38 sites from 9 coffee plantations of the Soconusco region of Chiapas, Mexico. While local management factors such as coffee density, branches per coffee bush, tree species, and density were not important in explaining leafhopper abundance and richness, shade management at the landscape level and elevation significantly affected leafhoppers. Specifically, the percentage of low-shade coffee in the landscape (1,000-m radius surrounding sites) increased total leafhopper abundance. In addition, Shannon's diversity of leafhoppers was increased with coffee density. Our results show that abundance and diversity of leafhoppers are greater in simplified landscapes, thereby suggesting that these landscapes will have higher pest pressure and may be more at-risk for diseases vectored by these species in an economically important crop.

Attraction of Coffee Bean Weevil, Araecerus fasciculatus, to Volatiles from the Industrial Yeast Kluyveromyces lactis.

The coffee bean weevil (CBW), Araecerus fasciculatus (De Geer, 1775) (Coleoptera: Anthribidae) is an important pest of stored products such as grains, coffee beans, cassava, and traditional Chinese medicine materials. In China, CBW causes large losses of Daqu, a traditional Chinese liquor fermentation starter, and, unfortunately, the use of conventional insecticides against CBW is not suitable in Daqu storage. We found CBW to be highly attracted to fermenting yeast cultures, such as Kluyveromyces lactis. Eight volatile compounds, produced by fermenting cultures and not by sterile samples, were identified by gas chromatography coupled with mass spectrometry. Five of these substances elicited significant responses in Y-tube behavioral bioassays. Field trapping experiments revealed 2-phenylethanol and 2-phenylethyl acetate to be crucial for attraction of CBW. Results show that yeast volatiles play an important role in host location, and that 2-phenylethanol and 2-phenylethyl acetate could be utilized as potential attractants in monitoring and control systems against this important pest.

A Keystone Ant Species Provides Robust Biological Control of the Coffee Berry Borer Under Varying Pest Densities.

Species' functional traits are an important part of the ecological complexity that determines the provisioning of ecosystem services. In biological pest control, predator response to pest density variation is a dynamic trait that impacts the provision of this service in agroecosystems. When pest populations fluctuate, farmers relying on biocontrol services need to know how natural enemies respond to these changes. Here we test the effect of variation in coffee berry borer (CBB) density on the biocontrol efficiency of a keystone ant species (Azteca sericeasur) in a coffee agroecosystem. We performed exclosure experiments to measure the infestation rate of CBB released on coffee branches in the presence and absence of ants at four different CBB density levels. We measured infestation rate as the number of CBB bored into fruits after 24 hours, quantified biocontrol efficiency (BCE) as the proportion of infesting CBB removed by ants, and estimated functional response from ant attack rates, measured as the difference in CBB infestation between branches. Infestation rates of CBB on branches with ants were significantly lower (71%-82%) than on those without ants across all density levels. Additionally, biocontrol efficiency was generally high and did not significantly vary across pest density treatments. Furthermore, ant attack rates increased linearly with increasing CBB density, suggesting a Type I functional response. These results demonstrate that ants can provide robust biological control of CBB, despite variation in pest density, and that the response of predators to pest density variation is an important factor in the provision of biocontrol services. Considering how natural enemies respond to changes in pest densities will allow for more accurate biocontrol predictions and better-informed management of this ecosystem service in agroecosystems.

Catechol--an oviposition stimulant for cigarette beetle in roasted coffee beans.

The cigarette beetle, Lasioderma serricorne, is a serious global pest that preys on stored food products. Larvae of the beetle cannot grow on roasted coffee beans or dried black or green tea leaves, although they oviposit on such products. We investigated oviposition by the beetles on MeOH extracts of the above products. The number of eggs laid increased with an increase in dose of each extract, indicating that chemical factors stimulate oviposition by the beetles. This was especially true for \ coffee bean extracts, which elicited high numbers of eggs even at a low dose (0.1 g bean equivalent/ml) compared to other extracts. Coffee beans were extracted in hexane, chloroform, 1-butanol, MeOH, and 20% MeOH in water. The number of eggs laid was higher on filter papers treated with chloroform, 1-butanol, MeOH, and 20% MeOH in water extracts than on control (solvent alone) papers. The chloroform extract was fractionated by silica-gel column chromatography. Nine compounds were identified by gas chromatography/mass spectrometry from an active fraction. Of these compounds, only a significant ovipositional response to catechol was observed.

Predictors of leafhopper abundance and richness in a coffee agroecosystem in Chiapas, Mexico.

Coffee agroecosystems with a vegetatively complex shade canopy contain high levels of biodiversity. However, as coffee management is intensified, diversity may be lost. Most biodiversity studies in coffee agroecosystems have examined predators and not herbivores, despite their importance as potential coffee pests and coffee disease vectors. We sampled one abundant herbivore group of leafhoppers on an organic coffee farm in Chiapas, Mexico. We sampled leafhoppers with elevated pan traps in high- and moderate-shade coffee during the dry and wet seasons of 2011. The two major objectives were to 1) compare leafhopper abundance and richness during the wet and dry seasons and 2) examine the correlations between habitat characteristics (e.g., vegetation, elevation, and presence of aggressive ants) and leafhopper richness and abundance. We collected 2,351 leafhoppers, representing eight tribes and 64 morphospecies. Leafhopper abundance was higher in the dry season than in the wet season. Likewise, leafhopper richness was higher in the dry season. Several vegetation and other habitat characteristics correlated with abundance and richness of leafhoppers. The number of Inga trees positively correlated with leafhopper abundance, and other significant correlates of abundance included vegetation complexity. Leafhopper richness was correlated with the number of Inga trees. As leafhoppers transmit important coffee diseases, understanding the specific habitat factors correlating with changes in abundance and richness may help predict future disease outbreaks.

The streamlined genome of Phytomonas spp. relative to human pathogenic kinetoplastids reveals a parasite tailored for plants.

Members of the family Trypanosomatidae infect many organisms, including animals, plants and humans. Plant-infecting trypanosomes are grouped under the single genus Phytomonas, failing to reflect the wide biological and pathological diversity of these protists. While some Phytomonas spp. multiply in the latex of plants, or in fruit or seeds without apparent pathogenicity, others colonize the phloem sap and afflict plants of substantial economic value, including the coffee tree, coconut and oil palms. Plant trypanosomes have not been studied extensively at the genome level, a major gap in understanding and controlling pathogenesis. We describe the genome sequences of two plant trypanosomatids, one pathogenic isolate from a Guianan coconut and one non-symptomatic isolate from Euphorbia collected in France. Although these parasites have extremely distinct pathogenic impacts, very few genes are unique to either, with the vast majority of genes shared by both isolates. Significantly, both Phytomonas spp. genomes consist essentially of single copy genes for the bulk of their metabolic enzymes, whereas other trypanosomatids e.g. Leishmania and Trypanosoma possess multiple paralogous genes or families. Indeed, comparison with other trypanosomatid genomes revealed a highly streamlined genome, encoding for a minimized metabolic system while conserving the major pathways, and with retention of a full complement of endomembrane organelles, but with no evidence for functional complexity. Identification of the metabolic genes of Phytomonas provides opportunities for establishing in vitro culturing of these fastidious parasites and new tools for the control of agricultural plant disease.

Cloning and biochemical characterization of an endo-1,4-ß-mannanase from the coffee berry borer Hypothenemus hampei.

BACKGROUND: The study of coffee polysaccharides-degrading enzymes from the coffee berry borer Hypothenemus hampei, has become an important alternative in the identification for enzymatic inhibitors that can be used as an alternative control of this dangerous insect. We report the cloning, expression and biochemical characterization of a mannanase gene that was identified in the midgut of the coffee berry borer and is responsible for the degradation of the most abundant polysaccharide in the coffee bean. METHODS: The amino acid sequence of HhMan was analyzed by multiple sequence alignment comparisons with BLAST (Basic Local Alignment Search Tool) and CLUSTALW. A Pichia pastoris expression system was used to express the recombinant form of the enzyme. The mannanase activity was quantified by the 3,5-dinitrosalicylic (DNS) and the hydrolitic properties were detected by TLC. RESULTS: An endo-1,4-ß-mannanase from the digestive tract of the insect Hypothenemus hampei was cloned and expressed as a recombinant protein in the Pichia pastoris system. This enzyme is 56% identical to the sequence of an endo-ß-mannanase from Bacillus circulans that belongs to the glycosyl hydrolase 5 (GH5) family. The purified recombinant protein (rHhMan) exhibited a single band (35.5 kDa) by SDS-PAGE, and its activity was confirmed by zymography. rHhMan displays optimal activity levels at pH 5.5 and 30°C and can hydrolyze galactomannans of varying mannose:galactose ratios, suggesting that the enzymatic activity is independent of the presence of side chains such as galactose residues. The enzyme cannot hydrolyze manno-oligosaccharides such as mannobiose and mannotriose; however, it can degrade mannotetraose, likely through a transglycosylation reaction. The K(m) and k(cat) values of this enzyme on guar gum were 2.074 mg ml(-1) and 50.87 s(-1), respectively, which is similar to other mannanases. CONCLUSION: This work is the first study of an endo-1,4-ß-mannanase from an insect using this expression system. Due to this enzyme's importance in the digestive processes of the coffee berry borer, this study may enable the design of inhibitors against endo-1,4-ß-mannanase to decrease the economic losses stemming from this insect.

Predatory potential of Euseius alatus (Phytoseiidae) on different life stages of Oligonychus ilicis (Tetranychidae) on coffee leaves under laboratory conditions.

This study evaluated the predatory capacity of Euseius alatus (DeLeon) as a biological control agent of the pest mite Oligonychus ilicis (McGregor) on coffee leaves under laboratory conditions, using arenas containing 25 O. ilicis per coffee (Coffea arabica) leaf to one specimen of each stage of the predator mite. The functional response and oviposition rate of adult females of E. alatus were evaluated on coffee leaf arenas and offered from 1 to 125 immature stages of O. ilicis per arena. The number of preys killed and the number of eggs laid by the predator were evaluated every 24 h during 8 days. The preys consumed were daily replaced. Male and female adults of E. alatus were the most efficient in killing all developmental stages of O. ilicis. Larvae and nymphs of O. ilicis were the most consumed by all stages of the predatory mite. The functional response and oviposition rates of E. alatus increased as the prey density increased, with a positive and highly significant correlation. Regression analysis suggested a type II functional response, with a maximum predation of 22 O. ilicis/arena and a maximum oviposition rate of 1.7 eggs/day at a density of 70 O. ilicis/arena.

Can volatile organic metabolites be used to simultaneously assess microbial and mite contamination level in cereal grains and coffee beans?

A novel approach based on headspace solid-phase microextraction (HS-SPME) combined with comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-ToFMS) was developed for the simultaneous screening of microbial and mite contamination level in cereals and coffee beans. The proposed approach emerges as a powerful tool for the rapid assessment of the microbial contamination level (ca. 70 min versus ca. 72 to 120 h for bacteria and fungi, respectively, using conventional plate counts), and mite contamination (ca. 70 min versus ca. 24 h). A full-factorial design was performed for optimization of the SPME experimental parameters. The methodology was applied to three types of rice (rough, brown, and white rice), oat, wheat, and green and roasted coffee beans. Simultaneously, microbiological analysis of the samples (total aerobic microorganisms, moulds, and yeasts) was performed by conventional plate counts. A set of 54 volatile markers was selected among all the compounds detected by GC×GC-ToFMS. Principal Component Analysis (PCA) was applied in order to establish a relationship between potential volatile markers and the level of microbial contamination. Methylbenzene, 3-octanone, 2-nonanone, 2-methyl-3-pentanol, 1-octen-3-ol, and 2-hexanone were associated to samples with higher microbial contamination level, especially in rough rice. Moreover, oat exhibited a high GC peak area of 2-hydroxy-6-methylbenzaldehyde, a sexual and alarm pheromone for adult mites, which in the other matrices appeared as a trace component. The number of mites detected in oat grains was correlated to the GC peak area of the pheromone. The HS-SPME/GC×GC-ToFMS methodology can be regarded as the basis for the development of a rapid and versatile method that can be applied in industry to the simultaneous assessment the level of microbiological contamination and for detection of mites in cereals grains and coffee beans.

Climate change or urbanization? Impacts on a traditional coffee production system in East Africa over the last 80 years.

Global environmental changes (GEC) such as climate change (CC) and climate variability have serious impacts in the tropics, particularly in Africa. These are compounded by changes in land use/land cover, which in turn are driven mainly by economic and population growth, and urbanization. These factors create a feedback loop, which affects ecosystems and particularly ecosystem services, for example plant-insect interactions, and by consequence agricultural productivity. We studied effects of GEC at a local level, using a traditional coffee production area in greater Nairobi, Kenya. We chose coffee, the most valuable agricultural commodity worldwide, as it generates income for 100 million people, mainly in the developing world. Using the coffee berry borer, the most serious biotic threat to global coffee production, we show how environmental changes and different production systems (shaded and sun-grown coffee) can affect the crop. We combined detailed entomological assessments with historic climate records (from 1929-2011), and spatial and demographic data, to assess GEC's impact on coffee at a local scale. Additionally, we tested the utility of an adaptation strategy that is simple and easy to implement. Our results show that while interactions between CC and migration/urbanization, with its resultant landscape modifications, create a feedback loop whereby agroecosystems such as coffee are adversely affected, bio-diverse shaded coffee proved far more resilient and productive than coffee grown in monoculture, and was significantly less harmed by its insect pest. Thus, a relatively simple strategy such as shading coffee can tremendously improve resilience of agro-ecosystems, providing small-scale farmers in Africa with an easily implemented tool to safeguard their livelihoods in a changing climate.

Molecular characterization of a miraculin-like gene differentially expressed during coffee development and coffee leaf miner infestation.

The characterization of a coffee gene encoding a protein similar to miraculin-like proteins, which are members of the plant Kunitz serine trypsin inhibitor (STI) family of proteinase inhibitors (PIs), is described. PIs are important proteins in plant defence against insects and in the regulation of proteolysis during plant development. This gene has high identity with the Richadella dulcifica taste-modifying protein miraculin and with the tomato protein LeMir; and was named as CoMir (Coffea miraculin). Structural protein modelling indicated that CoMir had structural similarities with the Kunitz STI proteins, but suggested specific folding structures. CoMir was up-regulated after coffee leaf miner (Leucoptera coffella) oviposition in resistant plants of a progeny derived from crosses between C. racemosa (resistant) and C. arabica (susceptible). Interestingly, this gene was down-regulated during coffee leaf miner herbivory in susceptible plants. CoMir expression was up-regulated after abscisic acid application and wounding stress and was prominent during the early stages of flower and fruit development. In situ hybridization revealed that CoMir transcripts accumulated in the anther tissues that display programmed cell death (tapetum, endothecium and stomium) and in the metaxylem vessels of the petals, stigma and leaves. In addition, the recombinant protein CoMir shows inhibitory activity against trypsin. According to the present results CoMir may act in proteolytic regulation during coffee development and in the defence against L. coffeella. The similarity of CoMir with other Kunitz STI proteins and the role of CoMir in plant development and plant stress are discussed.

Effective sampling range of a synthetic protein-based attractant for Ceratitis capitata (Diptera: Tephritidae).

Studies were conducted in Honduras to determine effective sampling range of a female-targeted protein-based synthetic attractant for the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Multilure traps were baited with ammonium acetate, putrescine, and trimethylamine lures (three-component attractant) and sampled over eight consecutive weeks. Field design consisted of 38 traps (over 0.5 ha) placed in a combination of standard and high-density grids to facilitate geostatistical analysis, and tests were conducted in coffee (Coffea arabica L.),mango (Mangifera indica L.),and orthanique (Citrus sinensis X Citrus reticulata). Effective sampling range, as determined from the range parameter obtained from experimental variograms that fit a spherical model, was approximately 30 m for flies captured in tests in coffee or mango and approximately 40 m for flies captured in orthanique. For comparison, a release-recapture study was conducted in mango using wild (field-collected) mixed sex C. capitata and an array of 20 baited traps spaced 10-50 m from the release point. Contour analysis was used to document spatial distribution of fly recaptures and to estimate effective sampling range, defined by the area that encompassed 90% of the recaptures. With this approach, effective range of the three-component attractant was estimated to be approximately 28 m, similar to results obtained from variogram analysis. Contour maps indicated that wind direction had a strong influence on sampling range, which was approximately 15 m greater upwind compared with downwind from the release point. Geostatistical analysis of field-captured insects in appropriately designed trapping grids may provide a supplement or alternative to release-recapture studies to estimate sampling ranges for semiochemical-based trapping systems.

Temperature-dependent development and emergence pattern of Hypothenemus hampei (Coleoptera: Curculionidae: Scolytinae) from coffee berries.

The coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae: Scolytinae), is the most important constrain for coffee production throughout the world. Knowledge on the emergence pattern of H. hampei females to infest new berries is crucial to effectively plan control measures. In this laboratory study, we assessed the development of immature stages and the emergence pattern of H. hampei females from the berries by exposing them to temperatures that are typical for high-altitude plantations (> or = 1,700 m above sea level [masl] ) or when coffee is grown under shade trees (20-22 degrees C), and optimum altitude plantations (1,200-1,600 masl) or nonshaded coffee (25-30 degrees C). Fecundity and emergence pattern of H. hampei females from coffee berries varied with temperature. Temperature played a crucial role determining the rate of H. hampei development and therefore the emergence of the females to start a new infestation cycle. The emergence and colonization phases of new colonizing females in coffee plantations with mean temperatures of 20, 25, or 30 degrees C would take place at different moments in the development of the coffee berries, and in some cases more than once. The implications of our findings for an improved, site-specific timing of control interventions against H. hampei are discussed.

Leaf alkaloids, phenolics, and coffee resistance to the leaf miner Leucoptera coffeella (Lepidoptera: Lyonetiidae).

Coffee (Coffea spp.) alkaloids (caffeine and related methylxanthines) and phenolics (caffeic and chlorogenic acids) have recognized pestistatic/pesticidal activity and mediate insect-plant interactions. The present investigation assessed the resistance of 12 coffee genotypes to the leaf miner Leucoptera (= Perileucoptera) coffeella (Guérin-Méneville & Perrottet) (Lepidoptera: Lyonetiidae) and correlated such results with the leaf content of coffee alkaloids and phenolics that probably play a role in the interaction between coffee and this leaf miner. The levels of chlorogenic and caffeic acid, caffeine, and related methylxanthines were measured and quantified in leaf extracts of these genotypes before and 7 d after their infestation by the leaf miner. Some coffee genotypes (Coffea canephora L. and Coffea racemosa Lour. and its hybrids with Coffea arabica L.) exhibited high pesticidal activity (100% mortality) toward the L. coffeella, indicating their antibiosis resistance. However, there was no correlation between this activity and the leaf levels of coffee alkaloids and phenolics. Curiously, infestation by L. coffeella leads to a nearly four-fold decline in the leaf levels of chlorogenic acid, which does not affect this pest species but may affect other generalist species. Indeed, chlorogenic acid sprayed on coffee leaves stimulated locomotory activity of the green scale Coccus viridis (Green) (Hemiptera: Coccidae), thus minimizing their feeding in contrast with the absence of this polyphenol. Therefore, reduction of chlorogenic acid levels in coffee leaves due to leaf miner infestation seems to also favor infestation by generalist insects, such as the green scale.