Use of advanced recombinant lines to study the impact and potential of mutations affecting starch synthesis in barley

The effects on barley starch and grain properties of four starch synthesis mutations were studied during the introgression of the mutations from diverse backgrounds into an elite variety. The lys5f (ADPglucose transporter), wax (granule-bound starch synthase), isa1 (debranching enzyme isoamylase 1) and sex6 (starch synthase IIa) mutations were introgressed into NFC Tipple to give mutant and wild-type BC₂F₄ families with different genomic contributions of the donor parent. Comparison of starch and grain properties between the donor parents, the BC₂F₄ families and NFC Tipple allowed the effects of the mutations to be distinguished from genetic background effects. The wax and sex6 mutations had marked effects on starch properties regardless of genetic background. The sex6 mutation conditioned low grain weight and starch content, but the wax mutation did not. The lys5 mutation conditioned low grain weight and starch content, but exceptionally high β-glucan contents. The isa1 mutation promotes synthesis of soluble α-glucan (phytoglycogen). Its introgression into NFC Tipple increased grain weight and total α-glucan content relative to the donor parent, but reduced the ratio of phytoglycogen to starch. This study shows that introgression of mutations into a common, commercial background provides new insights that could not be gained from the donor parent.     

The Effect of Consumption of Selenium Enriched Rye/Wheat Sourdough Bread on the Body’s Selenium Status

The potential of selenium-enriched rye/wheat sourdough bread as a route for supplementing dietary selenium intakes is reported. In addition to their normal diets, 24 female volunteers (24 to 25 years old) were fed either selenium-enriched bread or non-enriched bread each day (68.02 and 0.84 μg selenium day⁻¹ respectively) for 4 weeks. The chemical form of the selenium in the bread had been characterised using HPLC-ICP-MS, which showed that 42% of the extractable selenium was present as selenomethionine. Plasma selenium levels and plasma platelet glutathione peroxidase (GPx1) activity were measured in the volunteers’ blood over a 6-week period. A statistically significant difference (p = 0.001) was observed in the mean percentage change data, calculated from the plasma selenium level measurements for the enriched and control group, over the duration of the study. A comparable difference was not observed for the platelet GPx1 activity (p = 0.756), over the same period. Two weeks after cessation of the feeding stage, i.e., at t = 6 weeks, the mean percentage change value for the selenium plasma levels in the enriched group was still significantly elevated, suggesting that the absorbed selenium had been incorporated into the body’s selenium reserves, and was then being slowly released back into the volunteers’ blood.     

Epibulbar melanoma in a foal

A case of epibulbar melanoma in a 6-month-old, gelded, chestnut Hanoverian foal is reported. The location and clinical appearance upon initial presentation led to the tentative diagnosis of staphyloma or a congenital mass of unknown origin. An attempt was made to surgically excise the mass under general anesthesia, but due to its infiltrative nature and intraoperative appearance, most, but not all was removed without compromising the integrity of the globe. Histopathological evaluation revealed a multinodular to packeted, poorly demarcated, unencapsulated, infiltrative exophytic melanocytic neoplasm composed of bundles and nests of plump spindloid to polygonal heavily pigmented epithelioid neoplastic cells interspersed with pigment-laden macrophages within a fine fibrovascular stroma. Upon examination after enucleation, neoplastic cells were found to infiltrate into the lateral cornea, sclera and the choroid. This is a unique case of an epibulbar melanoma with choroidal invasion in a foal. Based on the sudden onset and rapid growth as well as the histological evidence of invasion, well-differentiated features, heavy pigmentation, and no apparent mitoses, this neoplasm was considered to be a low-grade malignant melanoma. At 14 months after excision there is no evidence of recurrence.     

Genetic regulation of conidiation in Trichoderma hamatun

Achieving a balance between vegetative growth and spore production is essential for successful biocontrol by fungi. Low sporulation rates in the field can result in poor establishment and survival,whereas failure of conidia to recognise hosts can lead to persistence without efficacy. Commercial biocontrol products involve bulk preparations of conidia, however considerable variability in conidiation rates exists between biocontrol agents, which can restrict choice of strain for production. The majority of studies on Trichoderma conidiation have focused on the species T. viride and T. atroviride.These species form conidia in response to blue and near-UV light and/or nutrient deprivation and conidiation proceeds in a highly co-ordinated fashion, however relatively little is known on the genetic basis of Trichoderrma conidiation. In addition, whilst photoconidiation appears to be a general response detailed studies in other Trichoderma species are absent. In this study, conidiation in the lesser known biocontrol species T. hamatum is being investigated using a combined morphological and molecular approach. In contrast to T. atroviride, conidiation in response to blue-light was weaker and variable and suggested that additional triggers may be required for the T. hamatum photoresponse. A series of comparative photoconidiation assays are currently being undertaken investigating the effect of inoculum type and abiotic factors on timing and intensity of the response.Results will be discussed in relation to the current knowledge on conidial morphogenesis in Trichoderma. In addition to these morphological assays, a selection of genes implicated in sporulation and the blue-light responses are currently being isolated and characterised from T. hamatum. Two genes, phr1 and cmp1 , which were isolated previously from T. atroviride will be used as early and late markers of gene expression during the photoresponse in T. hamatum in order to define time points for harvesting comparable stage-specific RNA from T. hamatum and T. atroviride. Using degenerate PCR putative sporulation gene orthologues have also been identified in T. hamatum.Work is currently underway to isolate genomic clones of these genes from T. hamatum and T.atroviride. Sequence and expression analysis of orthologues, including expression in response to abiotic factors will be presented and discussed in relation to the current knowledge of the molecular basis of conidiation in Trichoderma and other filamentous fungi.……     

An experimental test of the habitat amount hypothesis reveals little effect of habitat area but transient or indirect effects of fragmentation on local species richness

Landscape Ecology - The habitat amount hypothesis (HAH) posits that local species richness is driven more by the amount of habitat in the surrounding landscape than by local patch size or habitat...     

The effects of maternal nutrition during the first 50 d of gestation on the location and abundance of hexose and cationic amino acid transporters in beef heifer uteroplacental tissues

We hypothesized that maternal nutrition during the first 50 d of gestation would influence the abundance of hexose transporters, SLC2A1, SLC2A3, and SLC2A5, and cationic amino acid transporters, SLC7A1 and SLC7A2, in heifer uteroplacental tissues. Angus-cross heifers (n = 43) were estrus synchronized, bred via artificial insemination, and assigned at breeding to 1 of 2 dietary intake groups (CON = 100% of requirements to achieve 0.45 kg/d of BW gain or RES = 60% of CON intake) and ovariohysterectomized on day 16, 34, or 50 of gestation (n = 6 to 9/d) in a completely randomized design with a 2 × 3 factorial arrangement of treatments. Uterine cross-sections were collected from the horn ipsilateral to the corpus luteum, fixed in 10% neutral buffered formalin, sectioned at 5 µm, and stained via immunofluorescence for transporters. For each image, areas of fetal membrane (FM; chorioallantois), luminal epithelium (ENDO), superficial glands (SG), deep glands (DG), and myometrium (MYO) were analyzed separately for relative intensity of fluorescence as an indicator of transporter abundance. Analysis of FM was only conducted for days 34 and 50. No transporters in target areas were influenced by a day × treatment interaction (P ≥ 0.06). In ENDO, all transporters were differentially abundant from days 16 to 50 of gestation (P ≤ 0.04), and SLC7A2 was greater (P = 0.05) for RES vs. CON. In SG, SLC7A1 and SLC7A2 were greater (P ≤ 0.04) at day 34 vs. day 16. In DG, SLC2A3 and SLC7A1 were greater (P ≤ 0.05) for CON vs. RES heifers; furthermore, SLC7A1 was greater (P < 0.01) at day 50 vs. days 16 and 34 of gestation. In MYO, SLC7A1 was greater (P < 0.01) for CON vs. RES and was greater (P = 0.02) at days 34 and 50 vs. day 16. There were no differences in FM (P ≥ 0.06). Analysis of all uterine tissues at day 16 determined that SLC2A1, SLC2A3, and SLC7A2 were all differentially abundant across uterine tissue type (P < 0.01), and SLC7A1 was greater (P = 0.02) for CON vs. RES. Analysis of all uteroplacental tissues at days 34 and 50 demonstrated that all transporters differed (P < 0.01) across uteroplacental tissues, and SLC7A1 was greater (P < 0.01) for CON vs. RES. These data are interpreted to imply that transporters are differentially affected by day of gestation, and that hexose and cationic amino acid transporters are differentially abundant across utero-placental tissue types, and that SLC7A1 is responsive to maternal nutritional treatment.     

Measurement of genetic and environmental variation in barley (Hordeum vulgare) grain hardness

In this study, we assessed a broad range of barley breeding lines and commercial varieties by three hardness methods (two particle size methods and one crush resistance method (SKCS—Single-Kernel Characterization System), grown at multiple sites to see if there was variation in barley hardness and if that variation was genetic or environmentally controlled. We also developed near-infrared reflectance (NIR) calibrations for these three hardness methods to ascertain if NIR technology was suitable for rapid screening of breeding lines or specific populations. In addition, we used this data to identify genetic regions that may be associated with hardness. There were significant (p<0.05) genetic effects for the three hardness methods. There were also environmental effects, possibly linked to the effect of protein on hardness, i.e. increasing protein resulted in harder grain. Heritability values were calculated at >85% for all methods. The NIR calibrations, with R² values of >90%, had Standard Error of Prediction values of 0.90, 72 and 4.0, respectively, for the three hardness methods. These equations were used to predict hardness values of a mapping population which resulted in genetic markers being identified on all chromosomes but chromosomes 2H, 3H, 5H, 6H and 7H had markers with significant LOD scores. The two regions on 5H were on the distal end of both the long and short arms. The region that showed significant LOD score was on the long arm. However, the region on the short arm associated with the hardness (hordoindoline) genes did not have significant LOD scores. The results indicate that barley hardness is influenced by both genotype and environment and that the trait is heritable, which would allow breeders to develop very hard or soft varieties if required. In addition, NIR was shown to be a reliable tool for screening for hardness. While the data set used in this study has a relatively low variation in hardness, the tools developed could be applied to breeding populations that have large variation in barley grain hardness.