Genetic Mapping of a new Hippo allele, HpoN.1.2, in Drosophila melanogaster.

Genetic screens provide a mechanism to identify genes involved with different cellular and organismal processes. Using a Flp/FRT screen in the Drosophila eye we identified mutations that result in alterations and de-regulation of cell growth and division. From this screen a group of undergraduate researchers part of the Fly-CURE consortium mapped and characterized a new allele of the gene Hippo, HpoN.1.2.

Impact of defoliation severity on photosynthesis, carbon metabolism and transport gene expression in perennial ryegrass.

Defoliation severity affects grass regrowth. The changes to biological processes affecting regrowth induced by severe defoliation are not fully understood, nor have they been investigated at a molecular level in field-grown plants. Field-grown perennial ryegrass (Lolium perenne L.) plants were defoliated to 20, 40 or 60mm during winter. Throughout regrowth, transcript profiles of 17 genes involved in photosynthesis and carbon metabolism or transport were characterised in stubble and lamina tissue. Although defoliation to 20mm reduced residual lamina area and stubble water-soluble carbohydrate reserves compared with plants defoliated to 40 or 60mm, net herbage regrowth was not reduced. Transcript profiles indicated a potential compensatory mechanism that may have facilitated regrowth. At the one-leaf regrowth stage, plants defoliated to 20mm had greater abundance of photosynthesis-related gene transcripts (rca, rbcS1, rbcS2, fba, fbp and fnr) and 20% greater stubble total nitrogen than plants defoliated to 60mm. A greater capacity for photosynthesis in outer leaf sheaths may be one potential mechanism used by severely defoliated plants to compensate for the reduced residual lamina area; however, this premise requires further investigation.

Plants modify biological processes to ensure survival following carbon depletion: a Lolium perenne model.

BACKGROUND: Plants, due to their immobility, have evolved mechanisms allowing them to adapt to multiple environmental and management conditions. Short-term undesirable conditions (e.g. moisture deficit, cold temperatures) generally reduce photosynthetic carbon supply while increasing soluble carbohydrate accumulation. It is not known, however, what strategies plants may use in the long-term to adapt to situations resulting in net carbon depletion (i.e. reduced photosynthetic carbon supply and carbohydrate accumulation). In addition, many transcriptomic experiments have typically been undertaken under laboratory conditions; therefore, long-term acclimation strategies that plants use in natural environments are not well understood. METHODOLOGY/PRINCIPAL FINDINGS: Perennial ryegrass (Lolium perenne L.) was used as a model plant to define whether plants adapt to repetitive carbon depletion and to further elucidate their long-term acclimation mechanisms. Transcriptome changes in both lamina and stubble tissues of field-grown plants with depleted carbon reserves were characterised using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The RT-qPCR data for select key genes indicated that plants reduced fructan degradation, and increased photosynthesis and fructan synthesis capacities following carbon depletion. This acclimatory response was not sufficient to prevent a reduction (P<0.001) in net biomass accumulation, but ensured that the plant survived. CONCLUSIONS: Adaptations of plants with depleted carbon reserves resulted in reduced post-defoliation carbon mobilization and earlier replenishment of carbon reserves, thereby ensuring survival and continued growth. These findings will help pave the way to improve plant biomass production, for either grazing livestock or biofuel purposes.

Validation of reference genes for quantitative RT-PCR studies of gene expression in perennial ryegrass (Lolium perenne L.).

BACKGROUND: Perennial ryegrass (Lolium perenne L.) is an important pasture and turf crop. Biotechniques such as gene expression studies are being employed to improve traits in this temperate grass. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) is among the best methods available for determining changes in gene expression. Before analysis of target gene expression, it is essential to select an appropriate normalisation strategy to control for non-specific variation between samples. Reference genes that have stable expression at different biological and physiological states can be effectively used for normalisation; however, their expression stability must be validated before use. RESULTS: Existing Serial Analysis of Gene Expression data were queried to identify six moderately expressed genes that had relatively stable gene expression throughout the year. These six candidate reference genes (eukaryotic elongation factor 1 alpha, eEF1A; TAT-binding protein homolog 1, TBP-1; eukaryotic translation initiation factor 4 alpha, eIF4A; YT521-B-like protein family protein, YT521-B; histone 3, H3; ubiquitin-conjugating enzyme, E2) were validated for qRT-PCR normalisation in 442 diverse perennial ryegrass (Lolium perenne L.) samples sourced from field- and laboratory-grown plants under a wide range of experimental conditions. Eukaryotic EF1A is encoded by members of a multigene family exhibiting differential expression and necessitated the expression analysis of different eEF1A encoding genes; a highly expressed eEF1A (h), a moderately, but stably expressed eEF1A (s), and combined expression of multigene eEF1A (m). NormFinder identified eEF1A (s) and YT521-B as the best combination of two genes for normalisation of gene expression data in perennial ryegrass following different defoliation management in the field. CONCLUSIONS: This study is unique in the magnitude of samples tested with the inclusion of numerous field-grown samples, helping pave the way to conduct gene expression studies in perennial biomass crops under field-conditions. From our study several stably expressed reference genes have been validated. This provides useful candidates for reference gene selection in perennial ryegrass under conditions other than those tested here.

Altering systemic acid-base balance through nutrition failed to change secondary sex ratio.

There is evidence that differences in either maternal blood pH or dietary mineral content can result in alterations in secondary sex ratio in mammals. Altering the proportions of certain dietary minerals is known to influence blood pH, offering a possible explanation for this effect of diet on secondary sex ratio. The present study was performed to investigate whether altering blood pH by manipulating the dietary cation-anion difference (DCAD) would alter secondary sex ratio. The DCAD is calculated (in mEq per 100 g dry matter) as the difference between metabolically strong cations (Na + K) and metabolically strong anions (Cl + S) in the diet. Three hundred female mice were randomly allocated to either a low or high DCAD ration for 3 weeks before coitus. Urine pH was monitored before beginning the experiment, as well as before and after the breeding period, as a proxy for blood pH. Mice on the low DCAD diet had a lower urine pH (mean (+/- s.d.) 6.0 +/- 0.1) than mice on the high DCAD diet (8.2 +/- 0.6), but DCAD did not affect the percentage of mice that became pregnant, the number of offspring per pregnant mouse or the sex ratio of the neonate group. These results suggest that blood pH alone does not alter sex ratio and that an altered systemic pH is not the reason for reported mineral-related variations in sex ratio.