Symptomatology, (Co)occurrence and Differential Diagnostic PCR Identification of 'Ca. Phytoplasma solani' and 'Ca. Phytoplasma convolvuli' in Field Bindweed.

Field bindweed (Convolvulus arvensis) is one of the major natural plant hosts and reservoirs of 'Candidatus Phytoplasma solani' ('Ca. P. solani'), the causal agent of plant diseases in diverse agricultural crops, including Bois noir (BN) disease of grapevine. Phylogenetically, the most closely related phytoplasma to 'Ca. P. solani', the 'Ca. P. convolvuli', induces disease in field bindweed that is known by its symptoms as bindweed yellows (BY). The occurrence, coinfection and symptoms association of the two phytoplasmas in shared host plants were the subject of this study. Specific primers for the amplification of the elongation factor Tu gene (tuf) were developed for the identification of 'Ca. P. convolvuli' (by conventional nested PCR), as well as primers for simultaneous detection of 'Ca. P. solani' and 'Ca. P. convolvuli' by duplex SYBR Green real-time PCR. Among symptomatic bindweed plants, 25 and 41% were infected with a single phytoplasma species, 'Ca. P. solani' and 'Ca. P. convolvuli', respectively, while 34% were infected with both phytoplasmas. None of the non-symptomatic control plants carried phytoplasma, while non-symptomatic plants from our previous epidemiological studies in BN-affected vineyards were confirmed to be infected solely with 'Ca. P. solani'. Stamp gene typing revealed Rqg50 and Rqg31 'Ca. P. solani' genotypes in plants coinfected with 'Ca. P. convolvuli', while three diverse genotypes (Rqg50, GGY and Rpm35) were identified in a single locality with symptomatic bindweeds infected solely with 'Ca. P. solani'. Variations in symptoms and their association with each of the phytoplasmas are described and documented. The symptom of bushy appearance could be single out as specific for 'Ca. P. convolvuli' infection, while occurrence of 'Ca. P. solani' could not be unequivocally associated with specific alterations in infected bindweeds. The results are discussed in the context of the epidemiological and ecological complexity of 'Ca. P. solani'-induced diseases and the relationship between the two phytoplasma relatives in shared host plant.

Customizing the spent coffee for Trichoderma reesei cellulase immobilization by modification with activating agents.

This study has explored the possibility to reuse the waste, spent coffee material for the cellulase enzyme immobilization. By the coffee surface modification with different activating agents, it was attempted to develop the convenient method for creation of a capable porous carrier for this purpose. Among the most common activating agents, glutaraldehyde, chlorine dioxide and hydrogen peroxide provided the most acceptable choice for the coffee surface modification. The changes that occurred on the coffee surface due to agents' treatment exposure were recorded by using of the FTIR spectra and SEM micrographs. The highest immobilization yield (55%) and immobilization efficiency (45%) were attained during 30min of the treatment time, by employing of 30% chlorine dioxide aqueous solution within 6mL/g activator/carrier ratio. The kinetic process was found to be predicted by the pseudo-second-order model. The cellulase immobilization onto the coffee surface provides an excellent base for increasing the enzyme availability to the substrate and enhancing the enzyme productivity, by offering the new perspectives to the industrial sector.

Utilization of spent coffee grounds for isolation and stabilization of Paenibacillus chitinolyticus CKS1 cellulase by immobilization.

This study has explored the feasibility of using spent coffee grounds as a good supporting material for the Paenibacillus chitinolyticus CKS1 cellulase immobilization. An optimal operational conditions in a batch-adsorption system were found to be: carrier mass of 12 g/L, under the temperature of 45 °C and no pH adjustments. The immobilization yield reached about 71%. An equilibrium establishment between the cellulase and the carrier surface occurred within 45 min, whereas the process kinetics may be predicted by the pseudo-second-order model. An immobilized cellulase preparation expressed very good avicelase activity, this reached up to 2.67 U/g, and revealed an improved storage stability property, compared to free enzyme sample counterpart. The addition of metal ions, such as K(+) and Mg(2+) did not affect positively immobilization yield results, but on the contrary, contributed to an improved bio-activities of the immobilized cellulase, thus may be employed before each enzyme application. The method developed in this study offers a cheap and effective alternative for immediate enzyme isolation from the production medium and its stabilization, compared to other carriers used for the immobilization.

Geometric morphometric study of geographic and host-related variability in Aceria spp. (Acari: Eriophyoidea) inhabiting Cirsium spp. (Asteraceae).

The russet mite, Aceria anthocoptes (Nalepa), is the only eriophyoid that has been recorded on Cirsium arvense (L.) Scop. It has been noted in several European countries and recently in the USA. In this study we explored the geographic and host-related variability of Aceria spp. inhabiting different Cirsium spp. We applied landmark-based geometric morphometric methods to study morphological variability of three body regions (ventral, coxigenital and prodorsal) of 13 Aceria spp. populations inhabiting five Cirsium spp. in Serbia (Europe) and four Cirsium spp. in Colorado (North America). Analyses of size and shape variation revealed statistically significant differences between Aceria spp. living on European native and North American native Cirsium spp., as well as between A. anthocoptes s.s. inhabiting European C. arvense and North American C. arvense. The coxigenital region was the most informative when considering inter-population shape differences. European Aceria spp. dwelling on Cirsium spp., including A. anthocoptes s.s. from C. arvense, are characterized by higher inter-population size and shape variability than their North American counterparts. This finding supports a Eurasian origin of A. anthocoptes, presumed to consist of a complex of cryptic taxa probably coevolved with host plants in the native environment. Morphological similarity among Aceria spp. inhabiting North American native Cirsium spp. may indicate that speciation of A. anthocoptes started relatively soon after the host shift to plants different from C. arvense in the invaded region.

Stability of the pyrethroid pesticide bifenthrin in milled wheat during thermal processing, yeast and lactic acid fermentation, and storage.

BACKGROUND: Pesticide residues have become an unavoidable part of food commodities. In the context of increased interest for food processing techniques as a tool for reducing pesticide residues, it is interesting to study the potential loss of pesticides during lactic acid and yeast fermentation. In the present paper the effect of fermentation by Lactobacillus plantarum and Saccharomyces cerevisiae and storage on 23 °C on bifenthrin in wheat was investigated. In addition, the effect of sterilisation (applied in order to avoid contamination with wild microorganism strains, i.e. to determine the individual effects of used strains) on bifenthrin degradation was tested as well. RESULTS: No significant loss of bifenthrin was observed during storage, or after the sterilisation. During the lactic acid fermentation, reduction within wheat fortified with 0.5 mg kg(-1) was 42%, while quite lower within samples fortified with 2.5 mg kg(-1) , maximum 18%. In contrast, bifenthrin concentration was not reduced during yeast fermentation, as the reduction in fortified samples was in the range of spontaneous chemical degradation during incubation period. CONCLUSION: Possible bifenthrin contamination in wheat, in amounts over the maximum residue limits, could not be reduced by sterilisation or by yeast fermentation, but lactic acid fermentation could be an effective tool for minimising residual contamination.