The prevailing view that insects lack endogenous enzymes for plant cell wall (PCW) digestion had led to the hypothesis that PCW digestion evolved independently in different insect taxa through the establishment of symbiotic relationships with microorganisms. However, recent studies reporting endogenous PCW-degrading genes and enzymes for several insects, including phylogenetically basal insects and closely related arthropod groups, challenge this hypothesis. Here, we summarize the molecular and biochemical evidence on the mechanisms of PCW digestion in insects to analyze its evolutionary pathways. The evidence reveals that the symbiotic-independent mechanism may be the ancestral mechanism for PCW digestion. We discuss the implications of this alternative hypothesis in the evolution of plant-insect interactions and suggest that changes in the composition of lignocellulolytic complexes were involved in the evolution of feeding habits and diet specializations in insects, playing important roles in the evolution of plant-insect interactions and in the diversification of insects.

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Novel endogenous cDNAs of beta-1, 4-endoglucanases (Oa-EGase I and Oa-EGase II) were cloned from the cerambycid beetle Oncideres albomarginata chamela. Oa-EGase I- and Oa-EGase II-deduced proteins and three-dimensional structures possess all features, including general architecture, signature motifs and catalytic domains, of glycosyl hydrolase families 5 and 45 (GHF5 and GHF45) and also share high levels of homology with other beetle cellulases. Total carboxymethylcellulase activity of O. a. chamela was 208.13 U/g of larvae. Phylogenetic analyses suggest that insect GHF5 and GHF45 are very ancient gene families and indicate, at least in the case of GHF5, that this family likely evolved from a common ancestor rather than, as is often reported, via horizontal gene transfer. Beetle GHF45 cellulases did not cluster with other metazoan cellulases. However, the presence of GHF45 cellulases in ancient molluscan taxa puts into question the hypothesis of horizontal gene transfer for the evolution of cellulases in animals.

Published methods to isolate DNA from insects are not always effective in xylophagous insects because they have high concentrations of phenolics and other secondary plant compounds in their digestive tracts. A simple, reliable and labor-effective cetyltrimethylammonium bromide-polyvinylpyrrolidone (CTAB-PVP) method for isolation of high quality DNA from xylophagous insects is described. This method was successfully applied to PCR and restriction analysis, indicating removal of common inhibitors. DNA isolated by the CTAB-PVP method could be used in most molecular analyses.

Plant defensins are antimicrobial peptides that exhibit mainly antifungal activity against a broad range of plant fungal pathogens. However, their actions against Candida albicans have not been extensively studied. The mRNA for gamma-thionin, a defensin from Capsicum chinense, has been expressed in bovine endothelial cells. The conditioned medium of these cells showed antifungal activity on germ tube formation (60-70% of inhibition) and on the viability of C. albicans (70-80% of inhibition). Additionally, C. albicans was not able to penetrate transfected cells. Conditioned medium from these cells also inhibited the viability (80%) of the human tumor cell line, HeLa.

Arabinogalactans (AGs) are structural polysaccharides of the plant cell wall. A small proportion of the AGs are associated with hemicellulose and pectin. Furthermore, AGs are associated with proteins forming the so-called arabinogalactan proteins (AGPs), which can be found in the plant cell wall or attached through a glycosylphosphatidylinositol (GPI) anchor to the plasma membrane. AGPs are a family of highly glycosylated proteins grouped with cell wall proteins rich in hydroxyproline. These glycoproteins have important and diverse functions in plants, such as growth, cellular differentiation, signaling, and microbe-plant interactions, and several reports suggest that carbohydrate components are crucial for AGP functions. In beneficial plant-microbe interactions, AGPs attract symbiotic species of fungi or bacteria, promote the development of infectious structures and the colonization of root tips, and furthermore, these interactions can activate plant defense mechanisms. On the other hand, plants secrete and accumulate AGPs at infection sites, creating cross-links with pectin. As part of the plant cell wall degradation machinery, beneficial and pathogenic fungi and bacteria can produce the enzymes necessary for the complete depolymerization of AGs including endo-β-(1,3), β-(1,4) and β-(1,6)-galactanases, β-(1,3/1,6) galactanases, α-L-arabinofuranosidases, β-L-arabinopyranosidases, and β-D-glucuronidases. These hydrolytic enzymes are secreted during plant-pathogen interactions and could have implications for the function of AGPs. It has been proposed that AGPs could prevent infection by pathogenic microorganisms because their degradation products generated by hydrolytic enzymes of pathogens function as damage-associated molecular patterns (DAMPs) eliciting the plant defense response. In this review, we describe the structure and function of AGs and AGPs as components of the plant cell wall. Additionally, we describe the set of enzymes secreted by microorganisms to degrade AGs from AGPs and its possible implication for plant-microbe interactions.

Colletotrichum lindemuthianum was able to grow and produce extracellular cellulolytic activity in a defined medium containing cellulose as the main carbon substrate. As measured either by the hydrolysis of 4-methylumbelliferyl-beta-D -cellotrioside or the release of glucose from carboxymethylcellulose, activity reached a peak after 13 days of incubation and then declined whereas growth markedly increased afterwards. Detection of glucose in carboxymethylcellulose hydrolysates suggested the concerted operation of endo-1,4-beta-glucanase, cellobiohydrolase (exo-1,4-beta-glucanase) and beta-glucosidase activities. The highest levels of cellulolytic activity were obtained in media supplemented with cellulose and glutamate. Other carbon and nitrogen sources markedly influenced growth and enzyme production. Oligonucleotides homologous to specific regions of the cellobiohydrolase-encoding cbhII gene from Trichoderma reesei were used to isolate a C. lindemuthianum cbhII-DNA fragment whose sequence revealed homologies of 98% and 92% with the nucleotide and the deduced amino acid sequences of the corresponding cbhII-DNA of T. reesei, respectively. RT-PCR and Southern blot analyses of total RNA samples obtained from cellulose-grown but not from glucose-grown mycelium revealed the expression of the corresponding cbhII transcript. The cbhII-cDNA fragment was cloned and sequenced.

The Colletotrichum genus has been considered as one of the top 10 fungal pathogens in molecular plant pathology based on their scientific and agrobiological importance. Although the genus contains species with different lifestyles, most of the Colletotrichum sp. are known by their hemibiotrophic strategy of infection/invasion causing anthracnose disease in many economically important crops. Hemibiotrophy includes two sequential stages of infection, biotrophy and necrotrophy, in a series of steps that involve the participation of different virulence factors. In this review, we present the current status of the knowledge of such factors reported in this genus and a list of related genes identified in Colletotrichum sp. genomes.