Phylogenetic,genetic, and phenotypic diversity of Pseudomonas syringae pv. syringae strains isolated from citrus blast and black pit in Tunisia

Citrus blast and black pit caused by Pseudomonas syringae pv. syringae (Pss) is the only bacterial disease reported in Tunisian Citrus orchards. The phylogenetic relationship between Pss strains was studied based on multilocus sequence analysis (MLSA), using partial sequences of housekeeping genes rpoD, rpoB, gyrB, cts, and pfk for 14 representative Pss Citrus strains, including the reference strain LMG5496. The MLSA revealed that the studied Tunisian Citrus strains are closely related to LMG5496 and cluster in phylogroup 02. Based on the cts gene, the majority of Citrus strains clustered in clades “a” and “b”. However, five strains were placed in a newly defined clade “g”. We describe the presence of six different type III secreted effectors (T3SEs). These were found with frequencies of 100% for the effector hopAN1 and the helper hrpK1, 65% for hopT1-2, and 14% for hopN1, hopR1, and hopQ1-2. Investigation of copper resistance showed that 67% of our Pss Citrus strains from Tunisia are resistant to copper sulphate in vitro, and the copper resistance genes copABCDR were detected in 23% of the strains. Our results present new data concerning the genetic diversity and phylogeny, presence of T3SEs, and copper resistance within the Pss populations that affect Citrus in Tunisia.     

Trypsin from the viscera of Bogue (<Emphasis Type="Italic">Boops boops</Emphasis>): isolation and characterisation

Trypsin from the viscera of Bogue (Boops boops) was purified to homogeneity by precipitation with ammonium sulphate, Sephadex G-100 gel filtration and Mono Q-Sepharose anion exchange chromatography, with an 8.5-fold increase in specific activity and 36% recovery. The molecular weight of the purified enzyme was estimated to be 23 kDa by SDS–PAGE and size exclusion chromatography. The purified trypsin appeared as a single band on native-PAGE and zymography staining. The purified enzyme showed esterase-specific activity on N-α-benzoyl-l-arginine ethyl ester (BAEE) and amidase activity on N-α-benzoyl-dl-arginine-p-nitroanilide (BAPNA). The optimum pH and temperature for the enzyme activity, after 10 min incubation, were pH 9.0 and 55°C, respectively, using BAPNA as a substrate. The trypsin kinetic constants K _m and k _cat on BAPNA were 0.13 mM and 1.56 s⁻¹, respectively, while the catalytic efficiency k _cat /K _m was 12 s⁻¹ mM⁻¹. Biochemical characterisation of B. boops trypsin showed that this enzyme can be used as a possible biotechnological tool in the fish processing and food industries.     

First serological investigation of peste-des-petits-ruminants and Rift Valley fever in Tunisia

This study, carried out between September 2006 and January 2007, is the first cross-sectional serological investigation of peste-des-petits-ruminants (PPR) and Rift Valley fever (RVF) in Tunisia. The objective was to assess the potential need to develop a dual, recombinant PPR–RVF vaccine and how such a vaccine might be utilised in Tunisia. An overall PPR seroprevalence of 7.45% was determined, a finding supported by the high specificity (99.4%) and sensitivity (94.5%) of the ELISA used. On assessment of the diversity and density of mosquitoes in the sampling area, four species of RVF-vectors of the genus Aedes and Culex were identified. However, no serological evidence of RVF was found despite the use of a highly sensitive ELISA (99–100%). Larger scale investigations are underway to confirm these findings and the continuation of the emergency vaccination program against these two diseases remains valid.     

Exogenous application of calcium silicate improves salt tolerance in two contrasting tomato (Solanum lycopersicum) cultivars

The aim of this study was to investigate the impact of application of calcium silicate and salinity singly, on plant growth and nutritional behavior and photosynthetic pigments of tomato. Application of sodium chloride (NaCl) induced significant reduction in plant development and growth parameters. Salt stress also led to an accumulation of sodium (Na⁺) and a decrease in potassium (K⁺) concentration. Reduction of chlorophyll and carotenoid in leaves were amongst other symptoms in salt-affected plants in 2 cultivars. Rio Grande was qualified as salt sensitive and Moneymaker as the salt tolerant genotype. Application of Silicon (Si) only improved plant behaviour as compared to control. Furthermore, Si induced ameliorative effects on the growth potential of NaCl stressed plants. This Si-ameliorative effect on plant varied depending on the considered cultivar and Si concentration. Based on these results, application of calcium silicate was suggested as an alternative way to ameliorate the harmful effects of salinity on tomato.     

Effect of growth stage on the content and composition of the essential oil and phenolic fraction of sweet marjoram (Origanum majorana L.)

The variation in the content and composition of Origanum majorana L. essential oil has been studied. Plant material has been harvested at four phenological stages (early vegetative, late vegetative, budding and full-flowering). Essential oil yield varied from 0.04 to 0.09% reached during the full-flowering stage. Analysis of the essential oils by GC and GC/MS revealed the presence of 38 components represented mainly by oxygenated monoterpenes (64.01–71.4%), monoterpene hydrocarbons (21.73–29.92%) and sesquiterpene hydrocarbons (1.47–4.05%). The main components were terpinen-4-ol (29.13–32.57%), cis-sabinene hydrate (19.9–29.27%), trans-sabinene hydrate (3.5–11.61%), γ-terpinene (2.11–8.20%), bornyl acetate (1.52–2.94%) and linalool (1.05–1.39%). On the other hand, phenolic contents varied from 2.706 to 6.834 mg/g of dry weight obtained during the later vegetative stage. RP-HPLC analysis of the methanolic extract of O. majorana L. dried aerial parts showed the predominance of phenolic acids during the early vegetative stage whereas flavonoids predominate during the other stages of growth. The main phenolic acids identified were trans-2-hydroxycinnamic, rosmarinic, vanillic, chlorogenic, gallic and cinnamic whereas the main flavonoids were amentoflavone, apigenin, quercetin, luteolin, coumarin and rutin. Results obtained showed that later vegetative stage is characterized by the highest contents of bioactive compounds and therefore it could be considered as the best stage for harvesting marjoram plants.     

Lipolytic activity levels and colipase presence in digestive glands of some marine animals

Studies on the digestive secretions in aquatic animals can elucidate certain aspects of their nutritive physiology. The aim of the present study was to compare the digestive lipase and phospholipase activities in ten marine species belonging to four classes following the taxonomic classification of marine organisms. All aquatic digestive tissues tested are equipped with lipase and phospholipase activities, assuming the hydrolysis of fat-rich food. The lipolytic activities determined in the pancreases of cartilaginous fishes were greater than those in bony fishes, molluscs and crustaceans. This finding might be explained by the strong digestive utilization of fat-rich macronutrients by these carnivorous fishes. A trend of activities and stabilities at different pH and temperatures for crude lipases and phospholipases from these aquatic animals suggests that the optimum pH and temperature for marine lipases are species dependent. Interestingly, the sardine caecal lipase and phospholipase were found to be mostly stable in a broad range of acidic pH values. The maximum activities of lipolytic enzymes from the hepatopancreases of Hexaplex trunculus (molluscs) and Carcinus mediterranus (crustaceans) were found to be 50 and 60?°C, respectively, whereas the optimal temperature of lipolytic enzymes for the other species was classically around 40?°C. Thermoactivity of molluscs' lipolytic preparations makes them potential candidates in industrial applications. Among digestive glands studied, only pancreas (cartilaginous fish) contained the classically known colipase. Regarded as the most primitive living jawed vertebrates, cartilaginous fishes represented by sharks and rays could be considered as the oldest vertebrates possessing a complex digestive system like that of mammals.