RAPD and ISSR molecular markers in <Emphasis Type="Italic">Olea europaea</Emphasis> L.: Genetic variability and molecular cultivar identification

Thirty Portuguese and eight foreign olive (Olea europaea L.) cultivars were screened using Random Amplified Polymorphic DNA (RAPD) and Inter-Simple Sequence Repeat (ISSR) markers. Twenty RAPD primers amplified 301 reproducible bands of which 262 were polymorphic; and 17 ISSR primers amplified 204 bands of which 180 were polymorphic. The percentage of polymorphic bands detected by ISSR and RAPD was similar (88 and 87%, respectively). The genetic variability observed was similar in the Portuguese and foreign olive cultivars. Seven ISSR and 12 RAPD primers were able to distinguish individually all 38 olive cultivars. Twenty specific molecular markers are now available to be converted into Sequence Characterised Amplified Region (SCAR) markers. Relationships among Portuguese and foreign cultivars is discussed.     

Electron paramagnetic resonance studies of manganese toxicity,tolerance, and amelioration with silicon in snapbean

Plant genotypes within species differ widely in tolerance to excess manganese (Mn) that may occur in acid soils, or in neutral or alkaline soils having poor aeration caused by imperfect drainage or compaction. However, Mn tolerance mechanisms in plants are largely unknown. Silicon (Si) is reported to detoxify Mn within plants, presumably by preventing localized accumulations of Mn associated with lesions on leaves. Because Mn is paramagnetic, electron paramagnetic resonance (EPR) spectroscopy, shows promise as a tool for characterizing toxic and non‐toxic forms of Mn in tolerant and sensitive plants. The objective of our study was to use EPR to: i) determine the chemical/ physical state of Mn in Mn‐tolerant and ‐sensitive snapbean cultivars; and ii) characterize the protective effects of Si against Mn toxicity. Manganese‐sensitive Wonder Crop 2 (WC) and Mn‐tolerant Green Lord (GL) cultivars of snapbean were grown at pH 5.0, in a greenhouse, in a modified Steinberg solution containing: Mn=0.05mg.L^‐1 (optimal); Mn=1.0mgL^‐1 (toxic); Mn=1.0 mg L^‐1 plus Si=4 mg L^‐1; and Mn=0.05 mg L^‐1 plus 4 mg Si L^‐1. All trifoliate leaf samples exhibited a 6‐line EPR signal that is characteristic of hexaaquo Mn²⁺. In both cultivars, a higher EPR Mn²⁺ signal‐intensity generally correlated with lower total leaf mass, higher total Mn concentrations and more pronounced symptoms of toxicity. Tolerance to excess Mn coincided with lower Mn²⁺ signal intensity. Silicon treatments ameliorated Mn toxicity symptoms in both genotypes, decreased total leaf Mn concentrations, and decreased EPR Mn²⁺ signal intensity. Results suggest that Mn toxicity is associated with reduced electron transport and accumulation of oxidation products in leaves. Amelioration of Mn toxicity by Si is regarded as connected with a reduction in this Mn‐induced process. Results indicated that EPR spectroscopy can be useful in investigating the biochemical basis for differential Mn tolerance in plants. The EPR observations might also help plant breeders in developing Mn‐tolerant cultivars.     

Synergistic effects of vesicular-arbuscular mycorrhizal fungi and diazotrophic bacteria on nutrition and growth of sweet potato (Ipomoea batatas)

Summary Sweet potatoes were micropropagated and then transplanted from axnic conditions to fumigated soil in pots in the greenhouse. Spores of Glomus clarum were obtained from Brachiaria decumbens or from sweet potatoes grown in soil infected with this fungus and with an enrichment culture of Acetobacter diazotrophicus. Three experiments were carried out to measure the beneficial effects of vesicular-arbuscular mycorrhizal (VAM) fungi-diazotroph interactions on growth, nutrition, and infection of sweet potato by A. diazotrophicus and other diazotrophs obtained from sweet potato roots. In two of these experiments the soils had been mixed with ¹⁵N-containing organic matter. The greatest effects of mycorrhizal inoculation were observed with co-inoculation of A. diazotrophicus and/or mixed cultures of diazotrophs containing A. diazotrophicus and Klebsiella sp. The tuber production was dependent on mycorrhization, and total N and P accumulation were increased when diazotrophs and G. clarum were applied together with VAM fungal spores. A. diazotrophicus infected aerial plant parts only when inoculated together with VAM fungi or when present within G. clarum spores. More pronounced effects on root colonization and intraradical sporulation of G. clarum were observed when A. diazotrophicus was co-inoculated. In non-fumigated soil, dual inoculation effects, however, were of lower magnitude. ¹⁵N analysis of the aerial parts and roots and tubers at the early growth stage (70 days) showed no statistical differences between treatments except for the VAM+Klebsiella sp. treatment. This indicates that the effects of A. diazotrophicus and other diazotrophs on sweet potato growth were caused by enhanced mycorrhization and, consequently, a more efficient assimilation of nutrients from the soil than by N₂ fixation. The possible interactions between these effects are discussed.     

Contents of carboxylic acids and two phenolics and antioxidant activity of dried portuguese wild edible mushrooms

The organic acids and phenolics compositions of nine wild edible mushrooms species (Suillus bellini, Tricholomopsis rutilans, Hygrophorus agathosmus, Amanita rubescens, Russula cyanoxantha, Boletus edulis, Tricholoma equestre, Suillus luteus, and Suillus granulatus) were determined by HPLC-UV and HPLC-DAD, respectively. The antioxidant potential of these species was also assessed by using the DPPH* scavenging assay. The results showed that all of the species presented a profile composed of at least five organic acids: oxalic, citric, malic, quinic, and fumaric acids. In a general way, the pair of malic plus quinic acids were the major compounds. Only very small amounts of two phenolic compounds were found in some of the analyzed species: p-hydroxybenzoic acid (in A. rubescens, R. cyanoxantha, and T. equestre) and quercetin (in S. luteus and S. granulatus). All of the species exhibited a concentration-dependent scavenging ability against DPPH*. T. rutilans revealed the highest antioxidant capacity.     

Title: priority areas for genetic conservation of Eremanthus erythropappus (DC.) MacLeish in Brazil

Genetic Resources and Crop Evolution - Eremanthus erythropappus (DC.) MacLeish, commonly known as candeia, is a threatened tree species that occurs in Seasonal Semideciduous Forests in Brazil. The...     

New beverages of lemon juice enriched with the exotic berries maqui, açaı́, and blackthorn: bioactive components and in vitro biological properties

Following previous research on lemon juice enriched with berries, the aim of this work was to design new blends based on lemon juice mixed with different edible berries of exotic and national origin: maqui ( Aristotelia chilensis (Molina) Stuntz), a?a?? ( Euterpe oleracea Mart.), and blackthorn ( Prunus spinosa L.). The phytochemical characterization of controls and blends was performed by HPLC-DAD-ESI/MS(n). Their antioxidant capacity against DPPH, superoxide, and hydroxyl radicals and hypochlorous acid and their potential to inhibit cholinesterases were also assessed. The profiling of the red fruits and lemon revealed a wide range of bioactive phenolics. The novel beverage based on lemon juice and maqui berry (LM) was the most interesting blend in terms of antioxidant capacity. Berry control samples displayed reduced effects on acetylcholinesterase and butyrylcholinesterase, the lemon juice control being always the most active. This activity was also remarkable for lemon-blackthorn (LB) and lemon-a?a?? (LA) blends, the last being the most effective inhibitor of cholinesterases among all samples. The results suggested that lemon juice enriched with berries could be of potential interest in the design of new drinks with a nutritive related function on health for chronic diseases.     

Transgenic RNAi reveals essential function for CTCF in H19 gene imprinting

The imprinted regulation of H19 and Insulin-like growth factor 2 expression involves binding of the vertebrate insulator protein, CCCTC binding factor (CTCF), to the maternally hypomethylated differentially methylated domain (DMD). How this hypomethylated state is maintained during oogenesis and the role of CTCF, if any, in this process are not understood. With the use of a transgenic RNA interference (RNAi)-based approach to generate oocytes with reduced amounts of CTCF protein, we found increased methylation of the H19 DMD and decreased developmental competence of CTCF-deficient oocytes. Our results suggest that CTCF protects the H19 DMD from de novo methylation during oocyte growth and is required for normal preimplantation development.