Molecular Identification of Trichoderma spp. in Garlic and Onion Fields and In Vitro Antagonism Trials on Sclerotium cepivorum

ABSTRACT Trichoderma species are non-pathogenic microorganisms that protect against fungal diseases and contribute to increased crop yields. However, not all Trichoderma species have the same effects on crop or a pathogen, whereby the characterization and identification of strains at the species level is the first step in the use of a microorganism. The aim of this study was the identification – at species level – of five strains of Trichoderma isolated from soil samples obtained from garlic and onion fields located in Costa Rica, through the analysis of the ITS1, 5.8S, and ITS2 ribosomal RNA regions; as well as the determination of their individual antagonistic ability over S. cepivorum Berkeley. In order to distinguish the strains, the amplified products were analyzed using MEGA v6.0 software, calculating the genetic distances through the Tamura-Nei model and building the phylogenetic tree using the Maximum Likelihood method. We established that the evaluated strains belonged to the species T. harzianum and T. asperellum; however it was not possible to identify one of the analyzed strains based on the species criterion. To evaluate their antagonistic ability, the dual culture technique, Bell’s scale, and the percentage inhibition of radial growth (PIRG) were used, evidencing that one of the T. asperellum isolates presented the best yields under standard, solid fermentation conditions.

Characterization of a Peruvian isolate of Metarhizium anisopliae var. acridum, a pathogen of grasshoppers

The objective of this study was to characterize the Peruvian isolate of Metarhizium anisopliae var. acridum, CG 863, obtained from the grasshopper Schistocerca interrita, a crop pest in Peru. The characterization was done by comparing this isolate with two other ones of M. anisopliae var. acridum, from Brazil and Australia, and with an isolate of M. anisopliae var. anisopliae. The three M. anisopliae var. acridum isolates had similar growth profiles in agar plates at 25°C and 37°C, and similar RAPD patterns according to the analysis of three primers. However, regarding these parameters and conidial size, these isolates were very distinct when compared to M. anisopliae var. anisopliae isolate. Bioassays indicated that the Peruvian isolate is as pathogenic as the Brazilian isolate against nymphs of Rhammatocerus schistocercoides.

Antibacterial activity of essential oils on Xanthomonas vesicatoria and control of bacterial spot in tomato

The objective of this work was to evaluate the effects of plant essential oils (EOs) on the growth of Xanthomonas vesicatoria, on bacterial morphology and ultrastructure, and on the severity of tomato bacterial spot. EOs from citronella, clove, cinnamon, lemongrass, eucalyptus, thyme, and tea tree were evaluated in vitro at concentrations of 0.1, 1.0, 10, and 100% in 1.0% powdered milk. The effect of EOs, at 0.1%, on the severity of tomato bacterial spot was evaluated in tomato seedlings under greenhouse conditions. The effects of citronella, lemongrass, clove, and tea tree EOs, at 0.1%, on X. vesicatoria cells were evaluated by transmission electron microscopy. All EOs showed direct toxic effect on the bacteria at a 10%-concentration in vitro. Under greenhouse conditions, the EOs of clove, citronella, tea tree, and lemongrass reduced disease severity. EOs of clove and tea tree, and streptomycin sulfate promoted loss of electron-dense material and alterations in the cytoplasm, whereas EO of tea tree promoted cytoplasm vacuolation, and those of citronella, lemongrass, clove, and tea tree caused damage to the bacterial cell wall. The EOs at a concentration of 0.1% reduce the severity of the disease.

Flour production from shrimp by-products and sensory evaluation of flour-based products

The objective of this work was to evaluate the production of flour using by-products (cephalothorax) obtained from the shrimp (Litopenaeus vannamei) industry, and to perform a sensory analysis of shrimp flour-based products. Physicochemical and microbiological analyses on fresh cephalothorax and on manufactured flour were performed, as well as the determination of cholesterol content of this flour, and the sensorial evaluation of soup and pastry made with this flour. By the microbiological analyses, no pathogenic microorganism was detected in the samples. Physicochemical analyses of flour showed high levels of protein (50.05%) and minerals (20.97%). Shrimp cephalothorax flour showed high levels of cholesterol. The sensory evaluation indicated a good acceptance of the products, with satisfactory acceptability index (81% for soup, and 83% for pastry), which indicates that shrimp cephalothorax in the form of flour has a potential for developing new products.