Optimization of the mineral composition of in vitro culture media

Plant tissue culture requires the optimization of growth media. The effect of eight different macronutrient formulations on the in vitro culture of Gatharanthus roseus nodes was investigated: the caulogenesis rate decreased with the overall ion concentration and the ammonium content of the growth medium. The methodology proposed for the optimization of the mineral composition of the growth medium is based upon the adjustment of the macronutrients contents to the plant requirements. Such an approach was used for the comparison between the effects of the optimized growth medium and those of the usual Murashige and Skoog solution in the growth of the multiple shoots issued from Solatium paludosum root calluses. The new formulation MH1 [with a diminution of the ionic concentration, nitrogen (N) supplied in the form of nitrate and calcium (Ca) in larger amounts than potassium (K)] doubled the biomass production and tripled the yield of solamargine, a secondary metabolite.     

Global analysis of Coffea canephora Pierre ex Froehner (Rubiaceae) from the Guineo-Congolese region reveals impacts from climatic refuges and migration effects

As the second species used for commercial coffee, evaluation of Coffea canephora Pierre ex Froehner population genetics is a challenging task for coffee breeding. This study examined the pattern of genetic variability and genetic relationships of cultivated and wild populations of C. canephora sampled across the Guineo-Congolese region of Africa and some improved populations maintained in field genebanks. A total of 293 individuals, sampled from 17 populations, were genotyped with 39 nuclear microsatellite markers. Genetic diversity and structure were investigated with both a model-based and a graphical approach; isolation by distance was also tested. Relationships between the diversity clusters are discussed with regard to differentiation due to several glacial refuges during the Last Glacial Maximum (LGM). High genetic diversity within C. canephora is confirmed with a mean number of alleles of 11.85 per marker, a mean gene diversity of 0.72 and a mean observed heterozygosity of 0.36. An overall structure of two main groups (Guinean and Congolese) subdivided in six subgroups (2 for the Guinean and 4 for the Congolese) was found, including one described for the first time in the Guinean group. A fine structure within the Guinean group was also newly detected. Genetic structure of C. canephora appears to be consistent with its geographic repartition at the continent scale. Structure of diversity was found in accordance with localizations of refuge zones during LGM and migration from this period. Results from this genetic structure study raise our capabilities to better manage and use the collections of genetics resources for breeding purposes. Those results will be used in future association studies to optimize the number of genotypes to be phenotyped.     

Evaluation of Compost Maturity,Hydrophysical and Physicochemical Properties: Indicators for Use as a Component of Growing Media

The cocomposting of agricultural waste is a new management priority in Tunisia. In this study, four composts were evaluated by comparing the changes in measured hydrophysical and physicochemical properties and phytotoxicity. The organic wastes used were almond shell (AS), sesame bark (SB), olive husk (OH), and green and wood wastes (GW and WW, respectively). Composts CI and CII were composed of AS/SB and OH/SB, respectively, at a ratio of 75/25 (wet weight basis). CIII consisted of OH, SB and CAS (coarsely-ground almond shell used as a bulking agent) at a ratio of 55/25/20. Finally, CIV was composed of 25%SB+9%CAS+18%GW+48%WW. The composts studied were characterized by basic pH and an electric conductivity (EC) value ranging from 1.6 to 2.4 mS/cm. The organic matter contents (OM) and C/N ratios of composts ranged from 20 to 46% and from 10 to 21%, respectively. Based on hydrophysical analyses, composts CI, CIII and CIV, containing AS, were shown to have a porosity and a water content of 10-26% and 10-20%, respectively. The phytotoxicity of composts was studied on the basis of cress seed germination. Results revealed that differences in properties are mainly related to the nature of composted waste and that some of these composts are compatible for use as constituents in growing media for horticultural soilless cultures.     

A new hydroxylated nonaprenylhydroquinone from the Mediterranean marine sponge Sarcotragus spinosulus

Chemical investigation of the Mediterranean sponge Sarcotragus spinosulus led to the isolation of a new hydroxylated nonaprenylhydroquinone, along with two known metabolites, hepta- and octaprenylhydroquinones. The structure of the new metabolite was assigned by extensive 1D and 2D NMR analyses and MS studies. The antileukemic effect of the three compounds towards the chronic myelogenous leukemia (CML) cells line K562 was also evaluated.     

Puroindoline-a and puroindoline-b interact with the Saccharomyces cerevisiae plasma membrane through different amino acids present in their tryptophan-rich domain

Puroindolines are two small, basic cysteine-rich proteins isolated from Triticum aestivum seeds and characterized by a tryptophan-rich domain. They form the molecular basis of wheat grain hardness and display antimicrobial activity that may contribute to plant defence. Their antimicrobial activity is presumed to be due to their hydrophobic tryptophan-rich domain. However, little is known about their mode of action and there is no in vivo evidence that the binding of puroindolines to membranes is mediated by their tryptophan-rich domain. In this study, using a yeast complementation assay, we showed that puroindolines interact with the Saccharomyces cerevisiae plasma membrane. By site-directed mutagenesis of their tryptophan-rich domain, we determined that two tryptophan residues (W41 and W44) are mandatory for interaction of puroindoline-a with the yeast membrane whereas interaction of puroindoline-b depends on lysine residues. These results highlight that other residues than tryptophan play a critical role in the interaction of puroindolines with membranes, and probably their affinity for lipids and antimicrobial activities.     

Establishment,survivorship, and growth of yellow birch seedlings after site preparation treatments in large gaps

The occurrence of yellow birch (YB) in the northeastern forests of North America is a source of concern. Several guidelines suggest ways to favor the regeneration of this species, including creating openings from small gaps to large clearcuts, coupled or not with a variety of soil preparation treatments. However, it is not clear if soil preparation treatment favors YB establishment by simply increasing the availability of good seedbed types, or by also altering competition pressure exerted by interfering species during the period of seedling establishment. For this study, large gaps (900 m²) were created in a 70-year old YB dominated forest, to which three treatments differing in soil disturbance intensities were applied (i.e., soil scarification (i) using a rake [raking]; or (ii) by dragging slash out to the edges of the gap [slash drag]; and (iii) limited to the passage of the harvesting equipment [control]). The proportion of seedbed types reportedly most favorable for YB establishment (i.e., mineral-dominated) increased as the severity of the site preparation increased. The opposite was observed for organic-dominated seedbeds. As a result, the stocking of YB seedlings increased with the severity of site preparation. However, YB stocking was deemed sufficient in all gaps to ensure future canopy dominance, even in the control treatment. Although YB seedlings generally achieved greater heights as site preparation intensity increased, it was clear that this did not reflect vigorous growth as, on average, greater heights coincided with greater seedling height–diameter ratios. At the seedbed level, height–diameter ratio was associated with an increase in surrounding competition pressure and an increase in the incidence of stem apical death (SAD), which in turn decreased height differences among seedbed types by the end of the study. At the gap level, this blurred the advantages of site preparation over a laisser-faire strategy. The incidence of SAD was greatest in the slash drag and the rake treatments. Consequently, we cannot say that intense soil scarification is worth the expenses, especially in stands where YB seed sources are abundant.     

Kalmia removal increases nutrient supply and growth of black spruce seedlings: An effect fertilizer cannot emulate

In Canada's eastern boreal forest, the stagnant growth of black spruce (Picea mariana (Mill.) BSP) seedlings is often observed in the presence of ericaceous shrubs such as Kalmia angustifolia L. Many mechanisms, including allelopathic interference, reduced soil N mineralization, soil enzymes inhibition, and direct resource competition have been proposed to explain poor spruce growth in the presence of Kalmia. However, the relative importance of direct competition versus indirect interference remains unclear. Our objective was thus to adequately isolate the “Kalmia effect” from other growth-limiting factors and to determine if removal of Kalmia also resulted in fundamental changes in the biochemical properties of the forest floor. By sampling plots established in 2000, we evaluated how Kalmia eradication and spot fertilization influenced soil nutrient availability, N mineralization rates, microbial basal respiration and biomass, as well as planted black spruce seedling growth, dimensions, and foliar nutrient concentrations 6 years later. We measured higher extractable-P, mineralizable-N, seedling dimensions and growth rates, as well as lower extractable-K, total-K, basal respiration and microbial biomass, in plots without Kalmia than in those where Kalmia had been maintained from 2000 to 2006. Our results thus confirmed that Kalmia eradication over 6 years not only improved the growth and nutrition of black spruce seedlings, but also resulted in fundamental changes in the biochemical properties of the forest floor. We demonstrated that along with direct competition for resources, Kalmia interferes indirectly with black spruce by modifying nutrient cycling and energy fluxes in soil. Higher indices of available C in plots with Kalmia corroborates that Kalmia tannins or rhizodeposition may reduce N mineralization by stimulating microbial immobilization, a relation that however needs to be confirmed with longer term laboratory incubations. Our results indicated that although it had a positive influence on seedling growth, the fertilization effect was confined to the first few years following treatment application, and failed to influence soil processes as did Kalmia eradication. Further monitoring will indicate if the increased litterfall in fertilized plots will eventually initiate a second wave of fertilizer-induced changes to soil processes, as observed in other ecosystems.     

The successional status of sugar maple (Acer saccharum), revisited

Two complementary experimental designs at two contrasting scales (landscape/long term; individual tree/short term) were used for an in-depth evaluation of the successional status of sugar maple (AS: Acer saccharum Marsh.). First, forest disturbances during the 20th century and composition were mapped for two landscapes in the Du Lièvre watershed of southern Quebec. Our results show that, as well as dominating stands in the absence of fire, AS often rapidly developed dominance after fire, especially in the south of our study area. Similarly, a majority of AS-dominated stands clearcut in 1928 continued to be AS-dominated 60 years later. Second, we examined AS seedlings planted under two very contrasting light regimes. AS seedlings showed a combination of traits particularly adapted to tolerate shade under a low light regime. However, owing to a surprisingly high phenotypic plasticity, AS also exhibited efficient development under high light. This suggests the classification of AS as a late-successional species should indeed be revised and that generalist or trans-successional would be a more appropriate designation for this species. We discuss the ramifications of such a status revision, with an emphasis on the implications for its silviculture.     

Effects of propiconazole on extra-cellular enzymes involved in nutrient mobilization during <Emphasis Type="Italic">Trametes versicolor</Emphasis> wood colonization

The effects of propiconazole on extra-cellular enzyme levels in Trametes versicolor have been investigated during wood colonization and degradation. The working hypothesis was that the biocide could alter metabolic pathways, which could lead to an alteration of extra-cellular enzyme production. In the presence of a propiconazole sub-lethal concentration, the wood degradation rate decrease concomitantly with the lag phase of fungal development observed during wood colonisation. The pattern of production of enzymes involved in polysaccharide degradation (β-glucosidases, glucuronidases, cellobiohydrolases), nitrogen (leucine aminopeptidase) and phosphorus (acid phosphatase) mobilization was only slightly altered in the presence of the biocide. In experiments performed in the presence of propiconazole, there was a strong induction of chitinases at the beginning of the colonization process. Addition of caffeine, a pleiotropic drug, which is also a chitinase inhibitor, together with propiconazole resulted in synergistic inhibition of the fungal growth. The implication of these results in the development of a new wood preservation strategy is discussed.