The sexual differentiation of Cannabis sativa L.: A morphological and molecular study

Summary Cannabis sativa L. is a dioecious species with sexual dimorphism occurring in a late stage of plant development. Sex is determined by heteromorphic chromosomes (X and Y): male is the heterogametic sex (XY) and female is the homogametic one (XX). The sexual phenotype of Cannabis often shows some flexibility leading to the differentiation of hermaphrodite flowers or bisexual inflorescences (monoecious phenotype). Sex is considered an important trait for hemp genetic improvement; therefore, the study of the mechanism of sexual differentiation is of paramount interest in hemp research. A morphological and molecular study of Cannabis sativa sexual differentiation has been carried out in the Italian dioecious cultivar Fibranova.Microscopic analysis of male and female apices revealed that their reproductive commitment may occur as soon as the leaves of the fourth node emerge; the genetic expression of male and female apices at this stage has been compared by cDNA-AFLP. A rapid method for the early sex discrimination has been developed, based on the PCR amplification of a male-specific SCAR marker directly from a tissue fragment.Five of the several cDNA-AFLP polymorphic fragments identified have been confirmed to be differentially expressed in male and female apices at the fourth node. Cloning and sequencing revealed that they belong to nine different mRNAs that were all induced in the female apices at this stage. Four out of them showed a high degree of similarity with known sequences: a putative permease, a SMT3-like protein, a putative kinesin and a RAC-GTP binding protein.     

Seed set,size, and development after 4x × 2x and 4x × 4x crosses in citrus

Summary As triploids produce seedless fruits the best production is investigated. Comparative studies of embryogenesis, seed set, size, and germination were carried out following 4x×2x and 4x×4x crosses. On the average, both crosses yielded similar numbers and proportions of fully developed seeds per fruit. Seed development and germination took place precociously in 4x×2x seeds as compared to 4x×4x ones. The size of 4x×2x seeds was about 1/6 to 1/3 the size of 4x×4x seeds; thus, they were distinguishable from one another on the basis of size. Similarly, it was found that 3x and 4x seeds could be identified based on size following 4x×2x+4x crosses. It was concluded that 4x×2x crosses would be more successful than 2x×4x crosses for producing triploids.     

Effects of cultivar and environment on phenolic acids in buckwheat

Summary Phenolic acid contents were determined in seeds from five buckwheat cultivars grown at three locations in western Canada for four years. Buckwheat contained 12–16 g/kg of total phenolic acids, about 3 g/kg of esterified phenolic acids, and 8–13 g/kg of etherified phenolic acids. The latter represented 70 to 79% of the total phenolics. Variation in phenolic acids was mainly due to cultivar, seasonal effects and their interaction, while growing location had no significant effect. Phenolic acid contents of buckwheat were independent of seed color and protein content. Esterified phenolic acids exhibited a negative significant association with etherified phenolic acids in buckwheat.     

Glyphosate tolerance in maize (Zea mays L.). 2. Selection and characterization of a tolerant somaclone

Summary The progeny of 104 regenerated maize plants were screened for tolerance to the safe broad-spectrum herbicide glyphosate during seed germination and early growth. Seven somaclones showed varying degrees of resistance to the application of the herbicide at 1.2 mM (0.1 kg a.i. in 400 1 ha^-1 of water). Plants capable of a normal growth following treatment with 2.4 mM (0.2 kg ha^-1) glyphosate at the three leaf stage were selfed, and their progeny analyzed. A family able to tolerate the exposure to glyphosate at 2.4 mM was isolated and shown to maintain a photosynthetic rate comparable with control after the application of the herbicide.The selfed progeny of the tolerant somaclone was characterized as to the properties of two targets of glyphosate, the shikimate pathway enzymes 5-enol-pyruvyl-shikimate-3-phosphate (EPSP) synthase and 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase. In vitro tests ruled out the possibility that the tolerance was due to altered forms of these enzymes. Families showed significant variability with regard to EPSP and DAHP synthase levels, measured at different stages during seedling growth; however, not even these traits were correlated with in vivo response to glyphosate. The possible role of other physiological processes in determining the increased tolerance to the herbicide is discussed.Abbreviations DAHP 3-deoxy-D-arabino-heptulosonate-7-phosphate - EPSP 5-enol-pyruvyl-shikimate-3-phosphate - CER carbon-exchange rate - ID₅₀ concentration causing 50% inhibition     

Evaluation of Surface Runoff Prediction by AnnAGNPS Model in a Large Mediterranean Watershed Covered by Olive Groves

The distributed parameter and continuous simulation Annualized Agricultural Non‐Point Source (AnnAGNPS) model was implemented in the watershed Anzur (Spain) covered by olive groves, to assess its prediction capability of surface runoff at the large watershed scale in semi‐arid conditions. A 5‐year database reporting hydrological, geomorphological and land use characteristics of the watershed allowed model implementation. Almost 180 surface runoff events were modelled by AnnAGNPS and compared with the corresponding observations through statistical indexes and grouping the runoff events in order to evaluate the model at different temporal scales (event, monthly and seasonal). AnnAGNPS evaluation showed that, in general, runoff was estimated by the default model with low accuracy at all the investigated time scales, likely, as a result of a simple representation of spatial variability. Calibration (by reducing initial curve numbers (CN) of the olive groves) provided more accurate and satisfactory predictions of event, monthly and seasonal runoff volumes with a low effort in the parameterisation approach. The best model performance was achieved at the event scale. The runoff prediction reliability may be attributable to the AnnAGNPS inaccuracy in adjusting CN values during the continuous simulation of the soil moisture conditions, because estimations of daily evapotranspiration values are quite realistic. Copyright © 2015 John Wiley & Sons, Ltd.     

Mapping Changes in Land Cover Composition and Pattern for Comparing Mediterranean Rangeland Restoration Alternatives

The limited amount of studies addressing the long‐term effectiveness of restoration actions to combat land degradation is a constraint for current landscape restoration planning and implementation. Remote sensing data provide a unique opportunity for gathering information on the spatial and temporal variability of restoration processes and may contribute to fill this knowledge gap. The aim of this study is to compare different management actions undertaken in Mediterranean degraded rangelands (i.e. dense pine plantations, thinned pine plantations, protection and passive management) by analysing landscape composition and pattern change assessed from object‐based classification of aerial photograph of 1954, 1977 and 2006. Results show a clear link between landscape composition and structure and the different restoration actions, allowing comparison between their spatial and temporal dynamics. All restoration actions promoted an increase in woody vegetation cover compared with non‐restored areas. However, only protection and passive management favoured the restoration of dense, complex and well‐connected patches of native Mediterranean maquis. The proposed methodology is discussed with respect to its applicability for future Mediterranean landscape restoration planning in a wider geographic and ecological perspective. Copyright © 2015 John Wiley & Sons, Ltd.     

Detection of Botryosphaeriaceae species within grapevine woody tissues by nested PCR, with particular emphasis on the Neofusicoccum parvum/N. ribis complex

Several species of Botryosphaeriaceae and Phaeomoniella chlamydospora are common agents of grapevine decline worldwide. Currently, the use of culture independent PCR based techniques for detection of Botryosphaeriaceae within grapevine tissues has been limited to Botryosphaeria dothidea. In the present study, two Botryosphaeriaceae specific nested PCR assays were developed. One with a narrow target range, to detect Neofusicoccum parvum and the closely related species complex (Neofusicoccum parvum/N. ribis sensu Pavlic et al. Molecular Phylogenetics and Evolution 51:259–268, 2009) and another, with a wider range, to detect all 17 species of Botryosphaeriaceae which have been reported as potential wood pathogens of grapevine. The effectiveness of these assays was validated in vivo on naturally infected wood samples collected from standing vines and dormant grafted rooted cuttings commercialized in Italy by different nurseries in different years. All samples were also screened by means of a previously published nested PCR assay specific for Phaeomoniella chlamydospora. It was found that: 1) propagation material may play an important role as source of primary inoculum, not only of Phaeomoniella chlamydospora, as previously reported, but also for members of the Botryosphaeriaceae, among which Neofusicoccum parvum, Botryosphaeria dothidea and Diplodia seriata are the most common, and 2) multiple infections by different species belonging to Botryosphaeriaceae and/or Phaeomoniella chlamydospora occur frequently both in standing vines and propagation material. This last finding supports the hypothesis that at least some of the non-specific symptoms of grapevine decline may be due to the presence of different pathogens within host tissues.     

Phenolic composition and antioxidant activity of aqueous infusions from Capparis spinosa L. and Crithmum maritimum L. before and after submission to a two-step in vitro digestion model

This study investigated the phenolic composition and antioxidant activities of aqueous infusions from wild-grown caper (Capparis spinosa L.) and sea fennel (Crithmum maritimum L.) from the Dalmatia region (Croatia) before and after their submission to an in vitro digestion process. HPLC/UV-vis-DAD/ESI-MS analysis of the caper infusion identified rutin, kaempferol 3-O-rutinoside, and isorhamnetin 3-O-rutinoside as dominant flavonoids in the matrix together with a series of cinnamoylquinic acid derivatives, whereas in the sea fennel aqueous infusion chlorogenic acid (5-caffeoylquinic acid), its isomers, and higher derivatives were identified as almost the sole class of phenolics. Both infusions exhibited good and dose-dependent antioxidant activity before in vitro digestion by the DPPH method, the β-carotene bleaching method, and copper-induced oxidation of human LDL. The amount of total phenolics (Folin-Ciocalteu assay) strongly decreased in digested samples (from 3.0 and 2.2% in caper and sea fennel infusions, respectively, to <1.0%), as did their antioxidant activity as measured by the three aforesaid methods. The results showed that the majority of phenolic compounds detected in both infusions are not stable under applied simulated gastrointestinal conditions and that the stability of these secondary metabolites strongly depends on the nature of the corresponding matrix.     

Model aging and oxidation effects on varietal, fermentative, and sulfur compounds in a dry botrytized red wine

From harvest until wine arrives to the consumer, oxygen plays a crucial role in the definition of the final aroma. In the present research, the effect of the model oxidative aging on a dry red Botrytis wine, such as Italian Amarone, was considered. Amarone wine was submitted to model oxidative aging and then analyzed with two different approaches (SPE-GC-MS and HS-SPME/GC-MS). The same sampling plan was adopted to study the model aging of the same Amarone wine in anaerobic conditions. The HS-SPME/GC-MS method was applied to investigate for the first time the effect of the oxidative aging on a vast number of fermentative sulfur compounds. This research highlighted peculiar evolutions for several volatile compounds. In particular, benzaldehyde showed a sensitive increment during the oxidative aging, with a rate much higher than that reported for non-Botrytis red wines. On the other hand, several sulfides (dimethyl sulfide, 3-(methylthio)-1-propanol, etc.) disappeared after just 15 days of oxidative aging. A wine oxidation marker such as 3-(methylthio)-propanal was not found in any of the oxidized wines; conversely methionol-S-oxide was tentatively identified. This evidence has not been mentioned in the literature. A possible involvement of grape withering process and Botrytis in these mechanisms was supposed: a dry red wine, produced from the same but without any grape withering process and Botrytis infection (e.g., Bardolino wine), was submitted to oxidative aging and analysis. This red wine showed an evolution similar to those reported in the literature for dry red wines but significantly different from the Amarone wine.