Phylogenetic characterisation of Onobrychis species with special focus on the forage crop Onobrychis viciifolia Scop.

Sainfoin (Onobrychis viciifolia Scop.) is a perennial forage legume that possesses beneficial properties in the context of sustainable agriculture. In order to initiate a pre-breeding programme, we have assembled a germplasm collection of O. viciifolia from many geographic regions, as well as a collection of Onobrychis species that might be crossed with O. viciifolia to improve its biological and agronomic properties. The objective of this study was to obtain DNA sequences from a representative sample of these accessions in order to characterise their genetic diversity and to assess the robustness of the Onobrychis taxonomy. Because of potential problems with copurification of tannins and polyphenols from Onobrychis leaf tissue, three methods for DNA extraction were assessed and the most appropriate one identified. DNA sequences were obtained for the nuclear internal transcribed spacer region and the trnH-psbA and trnT-trnL intergenic spacers of the chloroplast genome. Neighbour joining trees were constructed and accessions were assigned to operational taxonomic units. The results indicated that there is substantial genetic diversity among Onobrychis species. Redundancies in species nomenclature were identified, as well as possible overlap between some of the sections into which the Onobrychis genus has been divided. A genetic distinction was apparent between O. viciifolia accessions from western Europe and those from eastern Europe and Asia, reflecting a similar division based on agronomic properties. Recognition of these different though related germplasms will be valuable in the design of breeding programmes for the rational improvement of sainfoin as a forage crop for sustainable agriculture.     

Cytological characterisation of the underutilized forage crop Onobrychis viciifolia Scop. and other members of the Onobrychis genus

Sainfoin (Onobrychis viciifolia Scop.) is a perennial forage legume that possesses beneficial properties in the context of sustainable agriculture. A germplasm collection of O. viciifolia was assembled from many geographic regions, as well as a collection of Onobrychis species that might be crossed with O. viciifolia to improve its biological and agronomic properties. The objective of the present study was to obtain cytological information for a broad range of O. viciifolia varieties and related species, in order to confirm and extend previous findings. The study set was 143 accessions of O. viciifolia as well as 34 accessions of 17 other species from the Onobrychis genus. Three of the O. viciifolia accessions were diploid (2n = 2x = 14) and the remainder tetraploid (2n = 4x = 28). Chromosomes were <5 μm in length. Among the other Onobrychis species, roughly half were found to be diploid and the other half tetraploid, with two species, O. arenaria (Kit.) DC. and O. alba subsp. laconica (Orph. ex Boiss.) Hayek, including a mixture of diploid and tetraploid accessions. The basic number of chromosomes was seven, except for O. aequidentata (Sm.) d’Urv. and O. crista-galli Lam., where it was eight. The 2C value for O. viciifolia was estimated at 2.5 pg. These cytological data will be valuable as part of a pre-breeding programme aimed at the eventual production of improved O. viciifolia varieties for use in sustainable agriculture.     

Has global change induced divergent trends in radial growth of Pinus sylvestris and Pinus halepensis at their bioclimatic limit? The example of the Sainte-Baume forest (south-east France)

? It is agreed that climate (precipitation and temperature) influences the distribution of plant species. Near the margins of a species’ natural range, climate becomes limiting to physiological processes. There, climate change may be expected to have a significant impact on tree growth and the species’ ranges may be altered.

? In order to assess what influence climate change could exert on the distribution of pine species at their margin, radial growth trends in ring-width chronologies over the last century were analysed. In the French Mediterranean area where climate change is characterized by increased temperature, forest plots were selected along an altitudinal transect on the north-facing slope of the Sainte-Baume mountain (Bouches-du-Rhône, France) where the ranges of Pinus sylvestris and Pinus halepensis overlap.

? Two growth patterns were identified. For P. halepensis, radial growth has increased at all altitudes indicating that climate change has improved growth conditions of this species near the margin of its ecological range. For P. sylvestris, radial growth has increased only at low altitudes and even decreased at high altitudes.

? It must be deduced that the growth changes observed cannot be generalised either at the species level or at the geographical level and must be interpreted with great caution.

     

A new bioclimatic model calibrated with vegetation for Mediterranean forest areas

? Water availability is one of the main factors explaining flora composition and growth in Mediterranean regions, where it may decline with climate change.

? Our goal was to develop a model for forest site assessment in Mediterranean environments, focusing on water availability to assess potential vegetation composition and productivity in any places, whatever their level of disturbance.

? We designed a statistical model, using global climatic and geographic variables, as well as detailed local topographic and edaphic variables, to compute a bioclimatic index for Mediterranean forest environments. This model was calibrated in France with a flora index from 325 old forests. The model explained 80.3% of the flora index variance. The method fills a gap in existing models, bridging scales from the region to forest sites.

? Beyond its theoretical aspect, it was designed to allow practical tools to be derived from it for decision-making and management, such as the assessment of climate change impact on vegetation, and of forest productivity. Its development and adaptation is possible in other Mediterranean regions, and in any region where water is one of the main limiting factors.

     

Natural products as sources for new mechanisms of herbicidal action

New mechanisms of action for herbicides are highly desirable to fight evolution of resistance in weeds, to create or exploit unique market niches, and to cope with new regulatory legislation. Comparison of the known molecular target sites of synthetic herbicides and natural phytotoxins reveals that there is little redundancy. Comparatively little effort has been expended on determination of the sites of action of phytotoxins from natural sources, suggesting that intensive study of these molecules will reveal many more novel mechanisms of action. Examples of natural products that inhibit unexploited steps in the amino acid, nucleic acid, and other biosynthetic pathways are given. AAL-toxin, hydantocidin, and various plant-derived terpenoids are discussed. Strategies and potential pitfalls of using natural products as leads for new herbicide classes are summarized.     

Detection of GTP-tubulin conformation in vivo reveals a role for GTP remnants in microtubule rescues

Microtubules display dynamic instability, with alternating phases of growth and shrinkage separated by catastrophe and rescue events. The guanosine triphosphate (GTP) cap at the growing end of microtubules, whose presence is essential to prevent microtubule catastrophes in vitro, has been difficult to observe in vivo. We selected a recombinant antibody that specifically recognizes GTP-bound tubulin in microtubules and found that GTP-tubulin was indeed present at the plus end of growing microtubules. Unexpectedly, GTP-tubulin remnants were also present in older parts of microtubules, which suggests that GTP hydrolysis is sometimes incomplete during polymerization. Observations in living cells suggested that these GTP remnants may be responsible for the rescue events in which microtubules recover from catastrophe.     

Exploration of plant genomes in the FLAGdb++ environment

Background

Efficient high throughput screening systems of useful mutants are prerequisite for study of plant functional genomics and lots of application fields. Advance in such screening tools, thanks to the development of analytic instruments. Direct analysis in real-time (DART)-mass spectrometry (MS) by ionization of complex materials at atmospheric pressure is a rapid, simple, high-resolution analytical technique. Here we describe a rapid, simple method for the genetic discrimination of intact Arabidopsis thaliana mutant seeds using metabolic profiling by DART-MS.

Results

To determine whether this DART-MS combined by multivariate analysis can perform genetic discrimination based on global metabolic profiling, intact Arabidopsis thaliana mutant seeds were subjected to DART-MS without any sample preparation. Partial least squares-discriminant analysis (PLS-DA) of DART-MS spectral data from intact seeds classified 14 different lines of seeds into two distinct groups: Columbia (Col-0) and Landsberg erecta (Ler) ecotype backgrounds. A hierarchical dendrogram based on partial least squares-discriminant analysis (PLS-DA) subdivided the Col-0 ecotype into two groups: mutant lines harboring defects in the phenylpropanoid biosynthetic pathway and mutants without these defects. These results indicated that metabolic profiling with DART-MS could discriminate intact Arabidopsis seeds at least ecotype level and metabolic pathway level within same ecotype.

Conclusion

The described DART-MS combined by multivariate analysis allows for rapid screening and metabolic characterization of lots of Arabidopsis mutant seeds without complex metabolic preparation steps. Moreover, potential novel metabolic markers can be detected and used to clarify the genetic relationship between Arabidopsis cultivars. Furthermore this technique can be applied to predict the novel gene function of metabolic mutants regardless of morphological phenotypes.     

Contraceptive effect and potential side-effects of deslorelin acetate implants in rats (Rattus norvegicus): Preliminary observations

During the last ten years, numerous species have been treated with deslorelin implants to induce contraception. The aims of the study were 1) to assess contraceptive efficacy of 4.7 mg subcutaneous deslorelin implants in rats, 2) to determine the latency of contraceptive effect, and 3) to determine potential side effects. Three experimental females were implanted and their estrous cycle was studied by vaginal smear. Two weeks after implantation, a male whose fertility was previously assessed with a control female, was introduced into their cage. No female conceived during the 4 mo following implantation. Additionally, 38 pet rats were recruited from clients in practice to test for potential side effects, including 6 males and 32 females with a mean age of 14 mo. Local reaction and transient weight gain during the first 2 wk, as well as behavioral changes were recorded. According to this pilot study, deslorelin implant could be used as a contraceptive method in female rats. The latency period is about 2 wk. Nevertheless, it might be possible to refine the treatment further using hormonal measurements. The duration of contraceptive effect is to be determined in an upcoming study.     

Joint action of natural and synthetic photosystem II inhibitors

The inhibitory action on photosystem II of four sorgoleone analogues, isolated from Sorghum bicolor (L) Moench, and two synthetic inhibitors, diuron and bentazone, was tested by measuring oxygen evolution of thylakoid membranes. The inhibition of oxygen evolution for mixtures of inhibitors was compared with the Additive Dose Model (ADM). ADM assumes that, at a defined response level, the effect of a mixture of inhibitors can be unambiguously expressed by the potency of either of the inhibitors applied separately. The slope of the logistic dose-response curves differed between the inhibitors; sorgoleone analogues had the steepest and bentazone the shallowest slope. The difference in slopes makes the interpretation of the isoboles less general and may reflect the differences in the interaction between the natural and the synthetic inhibitors with the binding site. The results suggest that there may be some limitation to ADM, namely that compounds with the same site of action might have different response curves if their mechanism of binding is different. The joint action of inhibitors follows ADM at I₅₀ . Therefore, the inhibitors can replace each other in any mixture ratio, based on the relative potencies of the pure inhibitors, without changing each other's effect on oxygen evolution. The joint action at I₂₀ and I₈₀ sometimes diverged from ADM. © 1999 Society of Chemical Industry     

Anhydrous goat's milk fat: thermal and structural behaviors studied by coupled differential scanning calorimetry and X-ray diffraction. 2. Influence of cooling rate

Crystallization and melting properties of triacylglycerols (TGs) in anhydrous goat's milk fat (AGMF) are investigated by X-ray diffraction as a function of temperature (XRDT) coupled with high-sensitivity differential scanning calorimetry (DSC), using synchrotron radiation and Microcalix. The polymorphic behavior of AGMF was monitored by varying the cooling rates between 5 and 1 degrees C/min from 45 to -20 degrees C with their subsequent melting at 1 degrees C/min. Quenching of AGMF at -20 degrees C was also examined to determine the metastable polymorphic form of AGMF. At intermediate cooling rates, TGs in AGMF crystallize, from about 18 degrees C in two different lamellar structures with triple chain length 3Lalpha stacking of 72 A and a double chain length 2Lalpha stacking of 48 A, which are correlated to two overlapped exothermic peaks recorded by DSC. A reversible transition sub alpha <--> alpha was observed. Subsequent heating at 1 degrees C/min shows numerous structural rearrangements before final melting. At fast cooling of AGMF (5 degrees C/min), similar unstable crystalline varieties are formed while three endotherms are recorded. Several new unstable lamellar structures are observed after quenching. All of these data are compared to those previously reported at slow cooling (0.1 degrees C/min) showing a relative stability of the structures formed. In spite of general similitude, the thermal and structural behavior of the goat's milk is more complex than that of the cow's milk.