Functional genomics of microspore embryogenesis

Isolated plant microspores, when stressed and cultured in vitro, can be diverted from their normal gametophytic pathway towards sporophytic development, with the formation of haploid embryos and ultimately doubled-haploid plants. This process is called androgenesis or microspore embryogenesis, and is widely used in plant breeding programmes to generate homozygous lines for breeding purposes. Protocols for the induction of microspore embryogenesis and the subsequent regeneration of doubled haploid (DH) plants have been successfully developed for more than 200 species. These practical advances stand in stark contrast to our knowledge of the underlying molecular genetic mechanism controlling this process. The majority of information regarding the genetic and molecular control of the developmental switch from gametophytic to sporophytic development has been garnered from four intensely studied (crop) plants comprising two dicotyledonous species, rapeseed (Brassica napus) and tobacco (Nicotiana tabacum), and two monocotyledonous species, wheat (Triticum aestivum) and barley (Hordeum vulgare). In these species the efficiency of microspore embryogenesis is very high and reproducible, making them suitable models for molecular studies. In the past, molecular studies on microspore embryogenesis have focussed mainly on the identification of genes that are differentially expressed during this developmental transition and/or early in embryo development, and have identified a number of genes whose expression marks or predicts the developmental fate of stressed microspores. More recently, functional genomics approaches have been used to obtain a broad overview of the molecular processes that take place during the establishment of microspore embryogenesis. In this review we summarise accumulated molecular data obtained in rapeseed, tobacco, wheat and barley on embryogenic induction of microspores and define common aspects involved in the androgenic switch.     

Grain weight responses to post-anthesis spikelet-trimming in an old and a modern wheat under Mediterranean conditions

Average grain weight is a major yield component contributing to its variation, especially in Mediterranean regions where grain weight is frequently exposed to terminal stresses affecting grain growth. Most of the literature agrees that wheat grain growth is hardly limited by the source. However, no source–sink ratios studies seem to have been conducted in the Mediterranean region to determine to what degree wheat grain growth is actually limited by the source in these particular regions. We conducted two field experiments in Catalonia (north-eastern Spain), where an old cultivar (Anza) and a more recently released one (Soissons) were sown in a range of different nitrogen and water availabilities and sowing dates. This was to analyse the degree of source limitation for grain growth. Sink size was modified by removing half of the spikelets c. 10 days after anthesis, virtually doubling the availability of assimilates per grain effectively growing.

Trimming the spikes did not produce significant changes in grain growth rate or duration of grain filling. Consequently, grain weight did not respond noticeably to the reduction in sink demand and any eventual response has been far from representing a strong competition among grains during grain filling.     

Development of an appropriate breeding scheme for tolerance to Empoasca kraemeri in common bean

Summary Significant advances in increasing tolerance to the leafhopper Empoasca kraemeri Ross & Moore in common bean have been obtained using a new breeding scheme where yield under leafhopper attack is the principal selection criterion in the evaluation of progenies. However, to further refine this breeding scheme, a study was conducted to determine whether selection for nonprotected yield would be more effective in early versus late generations. Two selection strategies were compared. In Strategy I, early generation selections in the F₂ and F₃ were compared to Strategy II where late generation selections were made in the F₄ and F₅, with the F₂ and F₃ generations advanced using single pod descent and bulk practices, respectively. Yield trials of the F₆ lines from both selection strategies were conducted under nonprotected and insecticide protected treatments. No significant differences were detected between the two selection strategies. However, Strategy II did produce advanced lines with greater nonprotected yields than did Strategy I, with the best F₆ line, in three out of four crosses, coming out of Strategy II. Late generation selection is recommended over early generation selection. Results of the new leafhopper breeding scheme, based on yield, are compared to the old breeding scheme where selections were made using visual selection practices in early generations.     

Seasonal Variations of Sulphate Reduction Rates, Sulphur Pools and Iron Availability in the Sediment of a Dystrophic Lagoon (Sacca di Goro, Italy)

The aim of this work was to analyse factors which regulate sulphide mobility in the sediments of a dystrophic lagoon (Sacca di Goro, the southern lagoon of the Po river delta, Italy). In 1995-96, sediment oxygen demand and variations in organic matter content, redox potential, iron availability, inorganic sulphur concentrations and sulphate reduction rates were measured in sediment profiles at three stations (G, 4 and 17) representative of the main areas of the lagoon. Stations differed mainly in their salinity range and primary producer communities. High concentrations of reactive iron (110-275 µmol ml-1) and low sulphate reduction rates (0.8-16.1 mmol m-2 d-1) were measured in the sediment. Moreover, high concentrations of reactive ferric iron were detected in winter and spring at the stations closest to the freshwater inputs. Nevertheless, in summer, high concentrations of free sulphides were detected in the porewater, although most of the reactive ferrous iron was not sulphide-bound, indicating that not all of the reactive iron pool was available to buffer against sulphide release and thus measures of reactive iron pools may not be a good measure of the true buffering capacity of the sediment. Furthermore, a considerable production of sulphide may occur in the decaying Ulva biomass in the water column, where its concentration will be independent of the potential buffering capacity of iron in the sediment. Therefore when assessing the vulnerability of coastal lagoons to dystrophic events, both the size and availability of the reactive iron pool as well as the site of sulphide production must be taken into consideration.     

8S globulin of mungbean [Vigna radiata (L.) Wilczek]: cloning and characterization of its cDNA isoforms, expression in Escherichia coli, purification, and crystallization of the major recombinant 8S isoform

Three isoforms of the cDNA of the major 8S globulin of mungbean, 8Salpha, 8Salpha', and 8Sbeta, were isolated, cloned, and characterized. The cDNA sequences of 8Salpha, 8Salpha', and 8Sbeta had open reading frames of 1362, 1359 or 1362, and 1359 bp, respectively, which code for 454, 453 or 454, and 453 amino acids corresponding to molecular weights of 51 973, 51 627 or 51 758, and 51 779, respectively. Homology in terms of cDNA and amino acid sequences was 91-92% between 8Salpha and 8Salpha', 87% between 8Salpha and 8Sbeta, and 86-88% between 8Salpha' and 8Sbeta. The signal peptide was found to be 1-25, 1-24 or 25, and 1-23 for 8Salpha, 8Salpha', and 8Sbeta, respectively, using the signalP website (Nielsen, H.; Engelbrecht, J.; Brunak, S.; von Heijne, G. Protein Eng. 1997, 10, 1-6). The propeptide was determined to be IVHREN. A single site for glycosylation (N-X-S/T) was observed about 90 amino acids from the C terminus. Homology between mungbean 8S isoforms and other 7-8S proteins ranged from 45 to 68% within members of the legume family and 29 to 34% for crops of different species. The major isoform 8Salpha was expressed in Escherichia coli and purified by successive ammonium sulfate fractionation, hydrophobic interaction, and Mono Q column chromatography. The recombinant 8Salpha, but not the native form, was successfully crystallized producing rhombohedral crystals.     

Occurrence and distribution of organochlorine pesticides (OCPs) in tomato (Lycopersicon esculentum) crops from organic production

Organochlorine pesticides (OCPs) were quantified by GC-ECD in tomato (Lycopersicon esculentum) during a vegetation period. Plants were harvested at 15, 60, and 151 days after seed germination. Leaves, stem, roots, and fruit (peel and flesh) were analyzed separately. The results showed that tomato plants were able to accumulate OCPs from soils, and a trend to reach the equilibrium among tissues at mature stages was also observed. Endosulfans comprised the main OCP group, probably due to its spray during summer months in the surrounding areas. Banned pesticides such as DDTs, heptachlor, and dieldrin were found. OCPs levels in the fruit were below the maximum residues limits (MRL) considered by the Codex Alimentarius. DDE/DDT and alpha-/gamma-HCH ratios of <1 would indicate recent inputs of DDT and lindane in the environment. The occurrence of OCPs in the study farm, where agrochemicals have never been used, is a result of atmospheric deposition of those pesticides.     

Leptin regulates striatal regions and human eating behavior

Studies of the fat-derived hormone leptin have provided key insights into the molecular and neural components of feeding behavior and body weight regulation. An important challenge lies in understanding how the rewarding properties of food interact with, and can override, physiological satiety signals and promote overeating. We used functional magnetic resonance imaging to measure brain responses in two human patients with congenital leptin deficiency who were shown images of food before and after 7 days of leptin replacement therapy. Leptin was found to modulate neural activation in key striatal regions, suggesting that the hormone acts on neural circuits governing food intake to diminish the perception of food reward while enhancing the response to satiety signals generated during food consumption.     

Rooted in grass: Challenging patterns of knowledge exchange as a means of fostering social change in a southeast Minnesota farm community

By convening a multidisciplinary team(the Monitoring Team) that included farmers,university and agency researchers, andnon-profit staff; a small group of farmers insoutheast Minnesota, U.S.A., bolstered thelegitimacy of the sustainable agriculturemovement. Through the experience of forming ateam and working with individuals who operatedwithin the mainstream knowledge paradigm,farmers gained validation of their knowledgeabout farming, while researchers came to valuealternative knowledge systems. In the contextof a socially embedded movement, farmers wereempowered by sharing their knowledge withresearchers, and ultimately contributed to thesustainable agriculture movement by challengingtraditional patterns of knowledge exchange.