Culture medium for improved production of conidia for identification and systematic studies of Fusarium pathogens.

Fusarium guttiforme and Fusarium ananatum are the etiological agents of fusariosis and fruitlet core rot in pineapple, respectively, producing mycotoxins that are harmful to the health of consumers. These two fungi are morphologically similar and difficulty in obtaining macroconidia of the species limits their identification. Different types of media are available for the culture of these pathogens, but not all of them favor F. ananatum and F. guttiforme macroconidia production. Therefore, the objective of this study was to develop a simple culture medium to improve rapid macro- and microconidia formation in both F. guttiforme and F. ananatum to facilitate taxonomic, pathogenicity and mycotoxin studies. In vitro analysis showed that basal medium with carboxymethyl cellulose (CMC) was better than other media tested with the highest macroconidia production at 7 days of incubation. The highest production of microconidia was with synthetic nutrient medium (SN) at 7 days. F. ananatum produced a relatively high number of microconidia with one septum in comparison to F. guttiforme when cultured in CMC, which suggests an additional character useful for Fusarium taxonomy. CMC medium may serve as an improved alternative to culture media currently used in Fusarium research and contribute to further knowledge of the taxonomy and mycotoxins of Fusarium species.

A Rapid and Reliable Method for Molecular Detection of Fusarium guttiforme, the Etiological Agent of Pineapple Fusariosis

Abstract Pineapple (Ananas comosus var. comosus) fusariosis is an economically important fungal disease affecting the plant and its fruit. A rapid and reliable diagnosis is the base of integrated disease management practices. Fusariosis has resulted in quarantines for pineapple products in Central America, Africa and Asia. Difficulties diagnosing and correctly identifying the fungus Fusarium guttiforme, agent of the pineapple fusariosis, have led to the search for new methodologies, and for this we developed a new reliable molecular method to detect it. For diagnostic purposes, real-time PCR of elongation factor gene 1-α (ef1) was used to rapidly, specifically and sensitively diagnose F. guttiforme. A pathogenicity test was conducted with slips of the pineapple cultivar Pérola, a multiplex PCR was run, and the results compared with those obtained with real-time PCR. The real-time PCR assay with its specific primer set could readity distinguish F. guttiforme from other Fusarium species known to occur on pineapple. The real-time PCR test had 95% sensitivity and 100% specificity with a significance level p<0.0001. For field samples the test had 100% sensitivity and specificity. Thus, this new test is fit for use in serial analyses of pineapples, and may have application in the evaluation of propagation materials and making quarantine decisions. The ability to rapidly and specifically detect F. guttiforme in plant samples will facilitate monitoring of the pathogen and improve disease management.

Genetically Modified Labeling Policies: Moving Forward or Backward?

One of the priorities to address food security is to increase the access of farmers to biotechnology, through the application of scientific advances, such as genetically modified organisms and food (GMF). However, the spread of (mis)information about their safety strengthens the clamor for mandatory GMF labeling. This paper provides an overview of food labeling policies, considering the principles suggested by the Codex Alimentarius Commission, and analyzes the consequences for the world food security of the Brazilian labeling policies compared to developed countries. We discuss the discriminatory application of GMF mandatory labeling in the absence of any scientific evidence as it has the potential of causing social harm and jeopardizes research, production, and distribution of food and consumers' right to information.

Fed-batch production of green coconut hydrolysates for high-gravity second-generation bioethanol fermentation with cellulosic yeast.

The residual biomass obtained from the production of Cocos nucifera L. (coconut) is a potential source of feedstock for bioethanol production. Even though coconut hydrolysates for ethanol production have previously been obtained, high-solid loads to obtain high sugar and ethanol levels remain a challenge. We investigated the use of a fed-batch regime in the production of sugar-rich hydrolysates from the green coconut fruit and its mesocarp. Fermentation of the hydrolysates obtained from green coconut or its mesocarp, containing 8.4 and 9.7% (w/v) sugar, resulted in 3.8 and 4.3% (v/v) ethanol, respectively. However, green coconut hydrolysate showed a prolonged fermentation lag phase. The inhibitor profile suggested that fatty acids and acetic acid were the main fermentation inhibitors. Therefore, a fed-batch regime with mild alkaline pretreatment followed by saccharification, is presented as a strategy for fermentation of such challenging biomass hydrolysates, even though further improvement of yeast inhibitor tolerance is also needed.

Green coconut mesocarp pretreated by an alkaline process as raw material for bioethanol production.

Cocos nucifera L., coconut, is a palm of high importance in the food industry, but a considerable part of the biomass is inedible. In this study, the pretreatment and saccharification parameters NaOH solution, pretreatment duration and enzyme load were evaluated for the production of hydrolysates from green coconut mesocarp using 18% (w/v) total solids (TS). Hydrolysates were not detoxified in order to preserve sugars solubilized during the pretreatment. Reduction of enzyme load from 15 to 7.5 filter paper cellulase unit (FPU)/g of biomass has little effect on the final ethanol titer. With optimized pretreatment and saccharification, hydrolysates with more than 7% (w/v) sugars were produced in 48h. Fermentation of the hydrolysate using industrial Saccharomyces cerevisiae strains produced 3.73% (v/v) ethanol. Our results showed a simple pretreatment condition with a high-solid load of biomass followed by saccharification and fermentation of undetoxified coconut mesocarp hydrolysates to produce ethanol with high titer.

Physical Characteristics of the Leaves and Latex of Papaya Plants Infected with the Papaya meleira Virus.

Sticky disease, which is caused by Papaya meleira virus (PMeV), is a significant papaya disease in Brazil and Mexico, where it has caused severe economic losses, and it seems to have spread to Central and South America. Studies assessing the pathogen-host interaction at the nano-histological level are needed to better understand the mechanisms that underlie natural resistance. In this study, the topography and mechanical properties of the leaf midribs and latex of healthy and PMeV-infected papaya plants were observed by atomic force microscopy and scanning electron microscopy. Healthy plants displayed a smooth surface with practically no roughness of the leaf midribs and the latex and a higher adhesion force than infected plants. PMeV promotes changes in the leaf midribs and latex, making them more fragile and susceptible to breakage. These changes, which are associated with increased water uptake and internal pressure in laticifers, causes cell disruption that leads to spontaneous exudation of the latex and facilitates the spread of PMeV to other laticifers. These results provide new insights into the papaya-PMeV interaction that could be helpful for controlling papaya sticky disease.

Comparison of biofilm and attachment mechanisms of a phytopathological and clinical isolate of Klebsiella pneumoniae Subsp. pneumoniae.

Some bacterial species can colonize humans and plants. It is almost impossible to prevent the contact of clinically pathogenic bacteria with food crops, and if they can persist there, they can reenter the human food chain and cause disease. On the leaf surface, microorganisms are exposed to a number of stress factors. It is unclear how they survive in such different environments. By increasing adhesion to diverse substrates, minimizing environmental differences, and providing protection against defence mechanisms, biofilms could provide part of the answer. Klebsiella pneumoniae subsp. pneumoniae is clinically important and also associated with fruit diseases, such as "pineapple fruit collapse." We aimed to characterize biofilm formation and adhesion mechanisms of this species isolated from pineapple in comparison with a clinical isolate. No differences were found between the two isolates quantitatively or qualitatively. Both tested positive for capsule formation and were hydrophobic, but neither produced adherence fibres, which might account for their relatively weak adhesion compared to the positive control Staphylococcus epidermidis ATCC 35984. Both produced biofilms on glass and polystyrene, more consistently at 40°C than 35°C, confirmed by atomic force and high-vacuum scanning electron microscopy. Biofilm formation was maintained in an acidic environment, which may be relevant phytopathologically.

Cell wall alterations in the leaves of fusariosis-resistant and susceptible pineapple cultivars.

Fusariosis, caused by the fungus Fusarium subglutinans f. sp. ananas (Syn. F. guttiforme), is one of the main phytosanitary threats to pineapple (Ananas comosus var. comosus). Identification of plant cell responses to pathogens is important in understanding the plant-pathogen relationship and establishing strategies to improve and select resistant cultivars. Studies of the structural properties and phenolic content of cell walls in resistant (Vitoria) and susceptible (Perola) pineapple cultivars, related to resistance to the fungus, were performed. The non-chlorophyll base of physiologically mature leaves was inoculated with a conidia suspension. Analyses were performed post-inoculation by light, atomic force, scanning and transmission electron microscopy, and measurement of cell wall-bound phenolic compounds. Non-inoculated leaves were used as controls to define the constitutive tissue characteristics. Analyses indicated that morphological differences, such as cell wall thickness, cicatrization process and lignification, were related to resistance to the pathogen. Atomic force microscopy indicated a considerable difference in the mechanical properties of the resistant and susceptible cultivars, with more structural integrity, associated with higher levels of cell wall-bound phenolics, found in the resistant cultivar. p-Coumaric and ferulic acids were shown to be the major phenolics bound to the cell walls and were found in higher amounts in the resistant cultivar. Leaves of the resistant cultivar had reduced fungal penetration and a faster and more effective cicatrization response compared to the susceptible cultivar.