Genome-Wide Insight into Profound Effect of Carbon Catabolite Repressor (Cre1) on the Insect-Pathogenic Lifecycle of Beauveria bassiana

Carbon catabolite repression (CCR) is critical for the preferential utilization of glucose derived from environmental carbon sources and regulated by carbon catabolite repressor A (Cre1/CreA) in filamentous fungi. However, a role of Cre1-mediated CCR in insect-pathogenic fungal utilization of host nutrients during normal cuticle infection (NCI) and hemocoel colonization remains explored insufficiently. Here, we report an indispensability of Cre1 for Beauveria bassiana’s utilization of nutrients in insect integument and hemocoel. Deletion of cre1 resulted in severe defects in radial growth on various media, hypersensitivity to oxidative stress, abolished pathogenicity via NCI or intrahemocoel injection (cuticle-bypassing infection) but no change in conidial hydrophobicity and adherence to insect cuticle. Markedly reduced biomass accumulation in the Δcre1 cultures was directly causative of severe defect in aerial conidiation and reduced secretion of various cuticle-degrading enzymes. The majority (1117) of 1881 dysregulated genes identified from the Δcre1 versus wild-type cultures were significantly downregulated, leading to substantial repression of many enriched function terms and pathways, particularly those involved in carbon and nitrogen metabolisms, cuticle degradation, antioxidant response, cellular transport and homeostasis, and direct/indirect gene mediation. These findings offer a novel insight into profound effect of Cre1 on the insect-pathogenic lifestyle of B. bassiana.

Description using ultramorphological techniques of the infection of Beauveria bassiana (Bals.-Criv.) Vuill. in larvae and adults of Atta sexdens (Linnaeus, 1758) (Hymenoptera: Formicidae)

Leaf-cutting ants, whose name comes from the habit of cutting leaves and other parts of plants, are dominant herbivores in the Americas, attacking different crops to keep symbiotic fungi (their main food source) in the colony. These ants are soil inhabitants, an environment rich in microorganisms, many of which are considered entomopathogenic. In order to describe a kind of microbial biological control of these ants through histological techniques, we used the fungus Beauveria bassiana (Bals.- Criv.) Vuill. Larvae and adult workers of Atta sexdens (Linnaeus, 1758) were exposed to a fungal suspension with the concentration of 106/conidia/mL. The specimens were adequately prepared to describe the behavior of the fungus on the integument at 24, 48 and 72 hours after initial exposure. Some specimens were used to represent these periods, being properly processed for scanning electron microscopy, which was applied to document the fungus development on the surface of the insect integument. To better understand the dynamics of the fungal infection, from the conidial adhesion to the potential colonization of the insides of the insect, the histological technique with HE staining was applied. The results showed that, in immature individuals, the hyphal penetration of B. bassiana fungi into the insect integument occurred in 48 h.

Entomopathogenic fungi as biological controllers: New insights into their virulence and pathogenicity

Entomopathogenic fungi vary considerably in their mode of action and virulence. Successful infection depends primarily on the adherence and penetration ability of a fungus to the host integuments. A variety of extracellular enzymes is produced during the degradation of insect integument. The attempts to control insects have changed over time from chemicals to natural control methods. This is why the development of natural methods of insect control or biopesticides, is preferred. By the use of fungal entomopathogens, insect pests can be controlled. There is no doubt that insects have been used for many years, but their effective use in the field remains elusive. However, their additional role in nature has also been discovered. Comparison of entomopathogens with conventional chemical pesticides depends on their efficiency and cost. In addition to efficiency, there are advantages in using microbial control agents, such as human safety and other non-target organisms; pesticide residues are minimized in food and biodiversity increased in managed ecosystems. In the present review the pathogenicity and virulence of entomopathogenic fungi and their role as biological control agents using biotechnology will be discussed.

Insect Epicuticular Grease Visualised by Scanning Probe Microscopy

The covering wings (elytra) in the Colorado potato beetle Leptinotarsa decemlineata appear shiny, smooth, water-repellent, and slightly slippery, (Fig. 1). These properties are due to the presence of epicuticle, the outermost layer of the insect integument, covered by a wax-like lipid surface layer called grease. Surface waxes have been previously reported in a variety of conditions, from liquid viscous coatings to crystalline structures in form of plates, rods and filaments from many insects (adults and larvae) and arachnids (Hadley, 1981a,b). Beament (1945) and Wigglesworth (1945) considered an outer thin layer of solid wax, whereas Lewis (1962) rather assumed an oil film on the epicuticle surface to be widespread throughout insects.

ISOLATION OF EPIDERMAL CELL PAIRS FROM AN INSECT, TENEBRIO MOLITOR, FOR DUAL WHOLE-CELL RECORDING OF LARGE-CONDUCTANCE GAP-JUNCTIONAL CHANNELS

The segmented insect integument, composed of an epidermal monolayer and its cuticular secretion, is a paradigm for the study of pattern formation during development (Lawrence, 1992). Epidermal activity during development may be coordinated by the transfer of cytoplasmic molecules through cell-cell gap-junctional channels (reviewed in Caveney, 1985). Gap junctions within intact epidermal segments exhibit dynamic changes in their permeability properties during the moult cycle in vivo (Caveney, 1978) and with exposure to the developmental hormone 20-hydroxyecdysone in vitro (Caveney and Blennerhassett, 1980). In addition, the row of epidermal cells at segment borders has distinct permeability properties creating developmental and communication compartments (Warner and Lawrence, 1982; Blennerhassett and Caveney, 1984). This paper documents a method for isolating epidermal cells that are suitable for dual whole-cell voltage-clamp studies of gap-junctional currents. We have identified a large-conductance gap-junctional channel in cell pairs with octanol- reduced junctional currents. This cell model may be useful for examining mechanisms of gap- junctional channel gating during development.

Neural regulation of pupariation in tsetse larvae.

A neural mechanism coordinates pupariation behavior and tanning in the tsetse larva. At parturition, the mature larva has already received sufficient ecdysteroid to commit the epidermal cells to metamorphosis but, before sclerotization and tanning of the cuticle can begin, the larva must first select a pupariation site and then proceed through a stereotypic sequence of pupariation behavior that culminates in the formation of a smooth, ovoid puparium. Both pupariation behavior and tanning are inhibited by the central nervous system (CNS) during the wandering phase. This central inhibition is maintained by sensory input originating in the extreme posterior region of the body. At the transition from wandering to pupariation, the posterior signal that induces inhibition of pupariation behavior is removed and the larva begins the contractions associated with pupariation, but the CNS inhibition of tanning persists. At this point, separation of the body into two halves by ligation or nerve transection prevents tanning of the anterior half (containing the CNS), whereas the denervated integument of the posterior half tans completely. Transection of nerves to the midline of the body produces larvae with a tanning pattern that ends abruptly along a sagittal plane, implying that the central control of this process is uncoupled between the left and right regions of the CNS. A few minutes later, when the final shape of the puparium is completed, the CNS inhibition is lifted and the tanning process begins. At this time, separation of the body into two halves by ligation or nerve transection has no inhibitory effects on either part. Exogenous ecdysteroids fail to release the CNS inhibition, and hemolymph containing the pupariation factors from Sarcophaga bullata have no accelerating effects on tsetse pupariation. These results imply that regulation of metamorphosis in the insect integument is not the exclusive domain of blood-borne hormones.