The parasitic nematode Metaparasitylenchus hypothenemi interferes with the endoparasitoid Phymastichus coffea,in the biological control of the coffee berry borer

We studied the interaction of Metaparasitylenchus hypothenemi with the exotic parasitoid Phymastichus coffea, when both natural enemies, co-ocurred inside the coffee berry borer adult. The nematode affects larvae and pupae of P. coffea, reducing by ten times the parasitoid’s adult progeny. We concluded that the negative effect caused by M. hypothenemi on the survival and development of juvenile stages of P. coffea could affect the action of this parasitoid as a natural enemy of this pest.     

Developments in coffee biotechnology—in vitro plant propagation and crop improvement

Coffee is an important plantation crop grown in about 80 countries across the globe. In recent years, coffee attained lot of attention in the biotechnology research area. Since last three decades, there has been a steady flow of information on coffee biotechnology and now it is entering into the genomic era. Major milestones in coffee biotech research are successful in vitro manipulation and multiplication of coffee, development of gene transfer protocols and generation of transgenic coffee plants with specific traits. The isolation of genes involved in caffeine biosynthetic pathway has opened up new avenues for generating caffeine free transgenic coffee. With the initiation of international coffee genomics initiatives, the genomic research in coffee is expected to reach new dimensions. The IPR issues may play crucial role in sharing of benefits during international collaborations in near future. This review focuses on the basic and applied aspects of coffee biotechnology for newer potentials.     

Effects of Management Intensity and Season on Arboreal Ant Diversity and Abundance in Coffee Agroecosystems

Agricultural intensification decreases arthropod predator diversity, abundance and population stability, and may affect interactions between top predators and their arthropod prey – ultimately affecting ecosystem services. Coffee management intensification (reduction or removal of shade trees) reduces diversity of arthropod predators (ground-foraging ants). Because ants provide ecosystem services by controlling pests, influences of intensification on arboreal, coffee-foraging ant diversity and abundance are important. We here address how coffee intensification affects: (1) coffee-foraging ant diversity and abundance and (2) seasonal fluctuations in ant abundance. In each of four coffee sites of varying management intensity in Chiapas, Mexico, we sampled vegetation and using two methods, sampled ant diversity and abundance over two years. Sites significantly differed in vegetation and management intensity. Coffee-foraging ant diversity generally decreased with increasing management intensity (16–26% fewer species observed in the most intensively-managed site). Ant abundance was higher in the wet season. Management intensity, however, did not influence ant abundance or seasonal fluctuations in abundance. Our results highlight the importance of diverse agricultural systems in maintaining arthropod predator diversity, and point to one model system in which we may effectively test how diversity per se affects ecosystem services.     

Regeneration filters in post-agricultural fields of Puerto Rico and Ecuador

To better understand how rainforest regenerates after agriculture I sampled the seed rain and seed bank, and set out seeds and seedlings, on microsites defined by distance to the forest in fields both in Puerto Rico and Ecuador. I found that (1) total seeds, species richness and life-form richness were twice as great in the Ecuador seed rain compared to Puerto Rico but Puerto Rico seeds were more evenly distributed among species and (2) total seedlings from the seed bank were similar between Puerto Rico and Ecuador, (3) the majority of seeds were lost to predation among all species and study sites, (4) seed disease was absent in P. aduncum and Miconia prasina, and no seeds germinated for Gonzalagunia spicata and P. riparia, (5) in Ecuador pathogenic disease claimed more seeds than germinated for all species, and Solanum ovalifolium was the only species that had seeds germinate but did not lose seeds to disease, (6) also in Ecuador, insect predation was significantly lower in the forest border for P. aduncum, and seed disease was significantly greater at the 10 m micro site for S. ovalifolium, (7) distance has a significant effect on seedling height and basal diameter, (8) losses of leaf area due to herbivory and pathogens were always low and (9) biomass and leaf specific mass were significantly reduced in the border and forest microsites. I conclude that Ecuador fields had more seeds, species, and life-forms than Puerto Rico fields, predation was the most severe post-dispersal seed filter in all fields, seeds that survived predation on Puerto Rico were lost either to disease or germination but to both mechanisms in Ecuador, all three seed mechanisms in Ecuador fields showed distance effects of seedling growth but not survivorship.     

Isolation,morphological and molecular characterization of Bacillus thuringiensis strains against Hypothenemus hampei Ferrari (Coleoptera: Curculionidae: Scolytinae)

The coffee berry borer Hypothenemus hampei Ferrari, 1876 (Coleoptera: Curculionidae: Scolytinae) is considered the most serious pest of the coffee crop and is controlled primarily with the use of chemical insecticides. An alternative to this control method is the use of the entomopathogenic bacterium, Bacillus thuringiensis Berliner, 1911. Therefore, the objective of this work was to select strains of B. thuringiensis virulent against H. hampei and characterize them by morphological and molecular methods to identify possible genes for the production of genetically modified plants. To achieve this objective, 34 strains of B. thuringiensis underwent a selective bioassay to evaluate their toxicity to H. hampei first-instar larvae. Among the strains tested, 11 and the standard B. thuringiensis subspecies israelensis (IPS-82) caused mortality above 90%. Then, the median lethal concentration (LC₅₀) was estimated for these strains followed by characterization using morphological, biochemical and molecular methods. The lowest LC₅₀ was obtained for strain BR58, although this concentration did not differ significantly from that of the standard strain IPS-82 or from that of strains BR137, BR80 and BR67. The molecular characterization detected cry4A, cry4B, cry10, cry11 and cyt1 genes in 10 of the most virulent strains (BR58, BR137, BR80, BR81, BR147, BR135, BR146, BR138, BR139, BR140). Strain BR67 differed completely from the others and amplified only the cry3 gene. This strain was more virulent than BR135, BR146, BR138, BR139 and BR140, but it did not differ from BR58, BR137, BR80, BR81 and BR147. The protein profile revealed proteins of 28, 65, 70 and 130 kDa, and the morphological analysis identified spherical crystalline inclusions in all strains. The results showed that the 11 strains studied have potential for use as a gene source for insertion into coffee plants for the control H. hampei, especially the cry3, cry4A, cry4B, cry10, cry11 and cyt1 genes, that were repeated in the most virulent isolates.     

Phylogenetic origins of African and Neotropical Beauveria bassiana s.l. pathogens of the coffee berry borer, Hypothenemus hampei

A phylogenetic epidemiological study of Beauveria bassiana s.l. was conducted for African and Neotropical pathogens of the coffee berry borer (CBB), Hypothenemus hampei, based on inferences from two nuclear intergenic regions, EFutr and Bloc. CBB pathogens were distributed among four terminal clades, however, the majority of African and Neotropical isolates cluster in a well-supported monophyletic group, informally designated AFNEO_1. Although the relationship between African and Neotropical AFNEO_1 is unresolved, the majority of alleles detected were exclusive to either the African or the Neotropical populations. These fixed genetic differences suggest that their disjunction predates the world trade in coffee. Neotropical AFNEO_1 have a broad host range and CBB pathogens are intermixed phylogenetically with isolates from diverse indigenous insects. Several Neotropical AFNEO_1 isolates were isolated from coffee plants as epiphytes or endophytes, thus plants themselves may potentially serve as reservoirs of pathogens against their insect pests. Topological incongruence between the EFutr and Bloc phylogenies of Neotropical AFNEO_1 may signify that individuals within this population are recombining.     

Human monoamine oxidase enzyme inhibition by coffee and beta-carbolines norharman and harman isolated from coffee

Monoamine oxidase (MAO) is a mitochondrial outer-membrane flavoenzyme involved in brain and peripheral oxidative catabolism of neurotransmitters and xenobiotic amines, including neurotoxic amines, and a well-known target for antidepressant and neuroprotective drugs. Recent epidemiological studies have consistently shown that coffee drinkers have an apparently lower incidence of Parkinson's disease (PD), suggesting that coffee might somehow act as a purported neuroprotectant. In this paper, "ready to drink" coffee brews exhibited inhibitory properties on recombinant human MAO A and B isozymes catalyzing the oxidative deamination of kynuramine, suggesting that coffee contains compounds acting as MAO inhibitors. MAO inhibition was reversible and competitive for MAO A and MAO B. Subsequently, the pyrido-indole (beta-carboline) alkaloids, norharman and harman, were identified and isolated from MAO-inhibiting coffee, and were good inhibitors on MAO A (harman and norharman) and MAO B (norharman) isozymes. beta-carbolines isolated from ready-to-drink coffee were competitive and reversible inhibitors and appeared up to 210 microg/L, confirming that coffee is the most important exogenous source of these alkaloids in addition to cigarette smoking. Inhibition of MAO enzymes by coffee and the presence of MAO inhibitors that are also neuroactive, such as beta-carbolines and eventually others, might play a role in the neuroactive actions including a purported neuroprotection associated with coffee consumption.     

Construction of a molecular linkage map in coffee

A linkage map for coffee (Coffea canephora P.) totalling 1402 cM has been developed on the basis of a population of doubled haploids. Both RFLP markers and PCR-based markers (RAPD) were used to construct 15 linkage groups. Coffee genomic and cDNA clones provided the source of the probes. In total, 47 RFLP and 100 RAPD loci have been placed on the linkage map. A rather low DNA polymorphism rate (18% for RFLP markers and 29% for RAPD primers) was detected. Only 81% of RAPD markers and 85% of RFLP markers fit an expected 11 ratio (P<0.01). The availability of a molecular linkage map has many implications for the future development of the genetics and breeding of this commercially important crop species.