Molecular biology of male gametogenesis

Pollen development has been studied at a molecular level in several systems that are amenable to genetic or transgenic analysis. We have characterized several tomato genes that are expressed late in pollen development. Our goals in this research were 1) to determine the cis- and tran-acting factors that mediate pollen expression, and 2) to determine the functions of the proteins encoded by these genes. We currently favor the hypothesis that pollen-specific gene expression is mediated in a combinatorial manner. Antisense experiments have indicated an important role for the LAT52 protein during pollen hydration.     

Methanolic extract of Verbascum macrurum as a source of natural preservatives against oxidative rancidity

The antioxidant properties of various fractions of a methanolic extract obtained from the aerial parts of Verbascum macrurum have been determined by monitoring their capacity to scavenge the stable free-radical DPPH. They were also evaluated as natural preservatives against oxidative rancidity using the accelerated Rancimat method. Their activities expressed as protection factor (PF(r)) indicated that the fractions rich with phenylpropanoid glycosides were more potent compared to alpha-tocopherol and of the same magnitude as BHT, which were used as reference standards. Ten natural compounds were identified as components of this methanolic extract and were isolated by medium-pressure liquid chromatography (MPLC). Assessment of their antioxidant activities established that acteoside, a polyhydroxylated phenylpropanoid glycoside derivative, is the most potent free radical scavenger and showed the highest protection factor (PF(r)) against sunflower-oil-induced oxidative rancidity. Its activity is comparable to the synthetic antioxidant BHT and clearly superior to natural alpha-tocopherol. This compound therefore represents a very interesting candidate for use in food preservation as natural protecting agent against oxidative rancidity.     

Comparative efficiencies of isopropyl and tert-butyl alcohols for extracting zeins from maize endosperm

Protein of endosperm of maize grains originating from three wild-type inbreds and their opaque-2 versions were solubilized in diverse extracts (E) by the sequential use of 0.5 M NaCl, water (E(1,2)), alcohol plus a reducing agent (E(3)), and salt plus a reducing agent (E(4)). Zeins were isolated in extracts E(3) and E(4) obtained by using 55% (w/w) isopropyl alcohol (i-PrOH) + 0.2% dithiothreitol (DTT) followed by 0.5 M NaCl + 0.2% DTT buffered at pH 10 or 60% tert-butyl alcohol (t-BuOH) + 0.2% DTT followed by 0.5% sodium acetate + 0.2% DTT in 30% t-BuOH. For a given genotype the percentage of extracted zeins was independent of the nature of the alcohol. The latter had a slight effect on the respective magnitude of E(3) and E(4): E(3) increased at the expense of E(4) when t-BuOH was substituted to i-PrOH for their isolation. The percentage of the total endosperm nitrogen present in E(3) + E(4) was identical to that of fractions F(II) + F(III) + F(IV) isolated according to the classical Landry-Moureaux extraction procedure. SDS-PAGE analysis revealed the presence of all types of zeins (alpha, beta, gamma, and delta) in E(3) and F(III), residual zeins in E(4) isolated with t-BuOH, and streaking only in E(4) and F(IV) isolated with NaCl at pH 10. The data together with those of the literature were discussed with regard to the influence of procedure on the yield of zeins using alcoholic extraction.     

Water usage and microclimate

Energy and water budget analyses are employed as methods for assessing the effects of vegetation type, cultivation practices, and irrigation methods on the microclimate. A comparison is made of vegetation types that employ water received from (1) natural precipitation and (2) irrigation. Forest lands are compared to corn cultivated by conventional tillage and no tillage methods. The forest canopy generally has a lower surface albedo, greater surface roughness, higher transpiration rates, and increased water storage than the bare soil, mulched, and vegetated surfaces of a corn field. No tillage cultivation reduces wind and water erosion, lowers soil temperatures, and improves water retention compared to standard tillage used on corn fields. Irrigated agriculture has replaced much of the drought resistant vegetation of the Central Valley of California. The energy and water budgets of irrigated crops grown in the semi-arid climate of the Central Valley and irrigated by flood or sprinkler methods exhibit microclimates in which evapotranspiration dominates the energy and mass fluxes. Drip irrigation methods, by contrast, have reduced water losses compared to flood and sprinkler methods. The drip system supplies metered amounts of water to the base of each plant; low soil evaporation and improved water supply to the growing plant results from this method. The latent heat flux is reduced over that found in fields irrigated by flooding or spraying. Generally, agricultural transformation of large stands of natural vegetation is expected to change the micro- and macroclimate of the areas affected.     

The effect of pH,aluminum, and chelator manipulations on the growth of acidic and circumneutral species of Asterionella

The growth rates of two diatoms, acidophilic Asterionella ralfsii and circumneutral A. formosa, were differentially affected by varying pH, Al, and EDTA in chemically defined media. Free Al ion concentration increased as pH and EDTA concentration decreased. Free trace metal ion concentration decreased as EDTA levels increased but increased by orders of magnitude upon addition of Al. pH had an overriding species specific effect on growth rate; at low pH A. ralfsii had higher growth rates than A. formosa and vice versa at high pH. For both species higher EDTA levels depressed growth rates. Moderate additions of Al generally resulted in growth stimulation. The growth rate stimulations, especially at 200 and 400 μg L^?1 Al additions, correlate to increases in free trace metal ion concentrations. The EDTA-AI interaction effects on growth rate were both pH and concentration dependent: at pH 7 both species were stimulated by addition of Al at all EDTA levels (except A. ralfsii at 5.0 mM EDTA and A. formosa at 0.5 mNM EDTA); at pH 6 Al addition either stimulated or had no effect on the growth rates of both species (except at low EDTA and high Al levels); at pH 5 A. formosa did not grow and additions of 200 μg L^?1 Al stimulated growth of A. ralfsii. It is likely that the effect of pH, Al, and EDTA on speciation of essential or toxic trace metals affects growth rates of these diatoms in a species specific manner.     

Do multipurpose companion trees affect high value timber trees in a silvopastoral plantation system?

Establishment of native timber trees on deforested land may contribute to the livelihood of farmers, to improved ecosystem services and to increased greenhouse gas uptake. Here, we present a new silvopastoral planting design to assess species performance and interspecific competition or facilitation effects among native timber and multipurpose trees in Central America. Two timber species, Tabebuia rosea and Cedrela odorata, were established in three low-density planting regimes allowing combined tree and future livestock production: (1) solitary planting, (2) companion planting with Guazuma ulmifolia, and (3) companion planting with the nitrogen-fixing Gliricidia sepium. We quantified survival, growth and reforestation potential of the two timber species subjected to the different planting regimes for the first 2 years after establishment. Nitrogen concentration as well as stable nitrogen and carbon isotope composition (δ¹⁵N, δ¹³C) of leaves of the timber saplings were determined. T. rosea showed higher survival and better growth than C. odorata under varying environmental conditions (soil, concomitant vegetation). Performance of the timber saplings was unaffected by either companion species. Planting regimes had no effect on foliar nitrogen concentration and δ¹⁵N of the two timber species, although δ¹⁵N values indicated nitrogen fixation activity in G. sepium trees. Planting regimes affected foliar δ¹³C values in T. rosea. δ¹³C values were significantly higher in solitarily growing individuals, suggesting lower exposition to water stress conditions in saplings surrounded by companion species. As we found positively correlated growth traits among timber and multipurpose trees, a combined planting may benefit farmers by providing additional goods and services.     

Temperature signals contribute to the timing of photoperiodic growth cessation and bud set in poplar

Bud set, the cornerstone delimiting the seasonal growth period in trees, is the dynamic net result of the often photoperiod-controlled growth cessation and the subsequent bud formation. Here, we show that in hybrid poplar, the critical day length for growth cessation and the duration of bud formation each vary with local climatic conditions in identical genotypes. The detailed dissection of bud set suggests temperature as one additional environmental factor that modifies the sensitivity to day-length signals at growth cessation and influences the duration of bud formation in poplar. The ability of perennial plants to integrate additional environmental signals with photoperiod signaling may add to short-term acclimatization to the predicted longer growing seasons in future climates.     

Establishment of native tropical timber trees in monoculture and mixed-species plantations: Small-scale effects on tree performance and insect herbivory

Plantations of native timber species established on former pasture are a potential strategy to reduce the logging pressure on remnant natural forests in the tropics. Such plantations may help to mitigate or reverse the negative impacts of land degradation, and they may contribute to the long-term livelihood of livestock farmers. Planting native trees is, however, perceived as a risky activity due to limited knowledge of their performance and due to marked losses of newly established seedlings attributed to insect pests. Our study focuses on the small-scale effects of environmental heterogeneity, tree diversity and insecticide treatment on the performance of three native Central American timber species two years after establishment, and on damage inflicted by insect herbivores. Growth, survival and herbivore leaf damage were quantified for Anacardium excelsum (Anacardiaceae), Cedrela odorata (Meliaceae) and Tabebuia rosea (Bignoniaceae) planted in an experiment in Panama as (1) monocultures, (2) mixed stands, and (3) mixed stands protected by insecticides. Our study revealed that small-scale effects can have a substantial impact on the success of timber trees planted on former pasture. Growth performance and survival of the three species was strongly affected by small-scale environmental heterogeneity, which was expressed as significant differences in growth and survival among different plantation plots at the same study site. Establishment of trees in mixed stands did not have significant effects on tree survival and growth compared to pure stands, although it reduced herbivore pressure in one of the studied tree species. All tree species grew best and suffered lowest leaf damage when protected by insecticides, indicating a general influence of insect herbivory on growth of newly established trees. In contrast to growth performance, survival was not significantly affected by different management practices or herbivory. The large variability among plots in tree survival and growth, and also in the effects of management practices such as planting design and insecticide treatment, emphasizes the importance of small-scale environmental heterogeneity on tree survival and growth.     

Carotenoid, chlorophyll, and chlorophyll-derived compounds in grapes and port wines

Carotenoids and chlorophyll-derived compounds in grapes and Port wines were investigated by HPLC-DAD and HPLC-DAD-MS (ESP+) analysis. A total of 13 carotenoid and chlorophyll-derived compounds are formally reported in grapes, 3 are identified for the first time, pheophytins a and b and (13Z)-beta-carotene, and 3 others remain unknown. In Port wines 19 compounds with carotenoid or chlorophyll-like structures are present, 8 still unidentified. The young wines showed higher total carotenoid content and chlorophyll-like compounds compared to aged Ports, with lutein and beta-carotene as major carotenoids. Among samples analyzed of monovarietal Vitis vinifera L. cultivar wines produced with the five most important Douro varieties, Tinta Roriz contained the highest levels of carotenoids and Touriga Franca the lowest. The forced-aging study indicated that lutein was more sensitive to temperature than beta-carotene. Additionally, aged wines showed higher ratios of beta-carotene/lutein concentrations compared to new Ports. Rates of degradation of chlorophyll derivative compounds were higher than those for carotene and lutein.     

In search of a maize ideotype for cell wall enzymatic degradability using histological and biochemical lignin characterization

Grass cell wall degradability is conventionally related to the lignin content and to the ferulic-mediated cross-linking of lignins to polysaccharides. To better understand the variations in degradability, 22 maize inbred lines were subjected to image analyses of Fasga- and M?ule-stained stem sections and to chemical analyses of lignins and p-hydroxycinnamic acids. For the first time, the nearness of biochemical and histological estimates of lignin levels was established. Combination of histological and biochemical traits could explain 89% of the variations for cell wall degradability and define a maize ideotype for cell wall degradability. In addition to a reduced lignin level, such an ideotype would contain lignins richer in syringyl than in guaiacyl units and preferentially localized in the cortical region rather than in the pith. Such enrichment in syringyl units would favor wall degradability in grasses, contrary to dicots, and could be related to the fact that grass syringyl units are noticeably p-coumaroylated. This might affect the interaction capabilities of lignins and polysaccharides.