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.     

Free amino acid composition of quince (Cydonia oblonga Miller) fruit (pulp and peel) and jam

Twenty-one free amino acids present in several samples of quince fruit (pulp and peel) and quince jam (homemade and industrially manufactured) were analyzed by GC/FID. The analyses showed some differences between quince pulps and peels. Generally, the highest content in total free amino acids and in glycine was found in peels. As a general rule, the three major free amino acids detected in pulps were aspartic acid, asparagine, and hydroxyproline. For quince peels, usually, the three most abundant amino acids were glycine, aspartic acid, and asparagine. Similarly, for quince jams the most important free amino acids were aspartic acid, asparagine, and glycine or hydroxyproline. This study suggests that the free amino acid analysis can be useful for the evaluation of quince jam authenticity. It seems that glycine percentage can be used for the detection of quince peel addition while high alanine content can be related to pear addition.     

Interactions of Glomus clarum with Acetobacter diazotrophicus in infection of sweet potato (Ipomoea batatas), sugarcane (Saccharum spp.), and sweet sorghum (Sorghum vulgare)

Summary Spores of the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus clarum obtained from sweet potatoes grown in soil inoculated with this fungus and with an enrichment culture of Acetobacter diazotrophicus contained A. diazotrophicus and several other bacteria, including a diazotrophic Klebsiella sp. Inoculation of micropropagated sweet potatoes with G. clarum and A. diazotrophicus enhanced spore formation in soil compared to VAM inoculation alone. Plants inoculated with VAM spores containing the bacteria showed additional increases in the number of spores formed within roots. A. diazotrophicus infected aerial plant parts only when inoculated together with VAM or when present within VAM spores. Micropropagated sugarcane seedlings inoculated with the same VAM spores containing the diazotrophs also contained much higher numbers of A. diazotrophicus in aerial parts than seedlings inoculated in vitro with the bacteria alone. When grown in non-sterile soil, the sugarcane seedlings again showed the greatest infection of aerial parts after inoculation with VAM spores containing the diazotrophs. This treatment also increased VAM colonization and the numbers of spores formed within roots. Similar effects were observed in sweet sorghum except that the aerial plant parts were not infected by A. diazotrophicus.     

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.     

Elevated atmospheric CO(2) affects the chemical quality of brassica plants and the growth rate of the specialist, Plutella xylostella, but not the generalist, Spodoptera littoralis

Cabbage, Brassica oleracea subsp. capitata (cv. Lennox and Rinda), and oilseed rape, Brassica rapa subsp. oleifera (cv. Valo and Tuli), plants were grown under ambient CO(2) (360 ppm) or elevated CO(2) (720 ppm) at 23/18 degrees C and under a photoperiod of 22/2 h light (250 micromol m(-)(2) s(-)(1))/dark regime for up to 5 weeks. Afterward, the performance of the crucifer specialist Plutella xylostella (Lepidoptera: Plutellidae) and the generalist Spodoptera littoralis (Lepidoptera: Noctuidae) on those plants was studied. The mean relative growth rate (RGR) of P. xylostella larvae, feeding on both cultivars of oilseed rape or on the Lennox cultivar of cabbage leaves grown at an elevated CO(2) concentration, was significantly reduced as compared to ambient CO(2). A negative larval growth rate at elevated CO(2) was observed for P. xylostella on both oilseed rape cultivars, but the growth rate was reduced but positive on cabbage. Conversely, the RGR of S. littoralis on either plant species was not affected by CO(2) treatment but was lower on cabbage cv. Rinda than on cv. Lennox. The mortality of the larvae was not affected by CO(2) treatment either. At the same time, elevated CO(2) significantly decreased the concentrations of leaf phytochemical constituents in oilseed rape, i.e., total phenolics and total nitrogen, but not in cabbage. The effect of elevated CO(2) on the leaf glucosinolate concentrations of both plant species was marginal. In addition, the observed significant changes in individual glucosinolate concentrations of oilseed rape leaves were not consistent among cultivars. However, our results demonstrate for the first time quite strong effects of CO(2) enrichment on the larval performance of P. xylostella, which is an important pest of Brassica plants around the world. Further studies are still required to increase our understanding of why elevated CO(2) differently affects the performance of specialist and generalist insect herbivores on Brassica plants.     

Field validation of chlordecone soil sequestration by organic matter addition

Purpose

The use of chlordecone (CLD) has caused pollution of soils, which are now a source of contamination for crops and ecosystems. Because of its long-term impacts on human health, exposure to CLD is a public health concern and contamination of crops by CLD must be limited. To this end, we conducted field trials on chlordecone sequestration in soil with added compost.

Materials and methods

The impact of added compost on chlordecone sequestration was measured in nitisols. After characterization of the soil, the transfer of chlordecone from soil to water was assessed in a leaching experiment and from soil to two crop plants in a nitisol plot. Finally, to understand the underlying processes, changes in CLD content were measured in soil fractions and soil porous properties were assessed after the addition of compost.

Results and discussion

A rapid seven-fold decrease in water extractable CLD was observed in amended soils. Five percent amendment led to a significant reduction in the contamination of crops by CLD; edible radish tubers were 50% more contaminated without added compost and cucumber fruits were 60% more contaminated. After the addition of compost, CLD content increased in the fraction of pre-humified or partially mineralized organic debris. Finally, in contrast to andosols, adding compost to nitisols did not affect the soil microstructure.

Conclusions

Increasing chlordecone sequestration by adding compost could be an alternative solution until soil decontamination techniques become available. This could be a provisional way to control further release of CLD from contaminated soils towards other environmental compartments.     

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.