Identification of Drosophila type II neuroblast lineages containing transit amplifying ganglion mother cells

Mammalian neural stem cells generate transit amplifying progenitors that expand the neuronal population, but these type of progenitors have not been studied in Drosophila. The Drosophila larval brain contains approximately 100 neural stem cells (neuroblasts) per brain lobe, which are thought to bud off smaller ganglion mother cells (GMCs) that each produce two post-mitotic neurons. Here, we use molecular markers and clonal analysis to identify a novel neuroblast cell lineage containing "transit amplifying GMCs" (TA-GMCs). TA-GMCs differ from canonical GMCs in several ways: each TA-GMC has nuclear Deadpan, cytoplasmic Prospero, forms Prospero crescents at mitosis, and generates up to 10 neurons; canonical GMCs lack Deadpan, have nuclear Prospero, lack Prospero crescents at mitosis, and generate two neurons. We conclude that there are at least two types of neuroblast lineages: a Type I lineage where GMCs generate two neurons, and a type II lineage where TA-GMCs have longer lineages. Type II lineages allow more neurons to be produced faster than Type I lineages, which may be advantageous in a rapidly developing organism like Drosophila.     

Allometric Models for Predicting Aboveground Biomass in Two Widespread Woody Plants in Hawaii

Allometric models are important for quantifying biomass and carbon storage in terrestrial ecosystems. Generalized allometry exists for tropical trees, but species‐ and site‐specific models are more accurate. We developed species‐specific models to predict aboveground biomass in two of the most ubiquitous natives in Hawaiian forests and shrublands, Metrosideros polymorpha and Dodonaea viscosa. The utility of the M. polymorpha allometry for predicting biomass across a range of sites was explored by comparing size structure (diameter at breast height vs. tree height) of the trees used to develop the models against trees from four M. polymorpha‐dominated forests along a precipitation gradient (1630–2380 mm). We also compared individual tree biomass estimated with the M. polymorpha model against existing generalized equations, and the D. viscosa model with an existing species‐specific model. Our models were highly significant and displayed minimal bias. Metrosideros polymorpha size structures from the three highest precipitation sites fell well within the 95% confidence intervals for the harvested trees, indicating that the models are applicable at these sites. However, size structure in the area with the lowest precipitation differed from those in the higher rainfall sites, emphasizing that care should be taken in applying the models too widely. Existing generalized allometry differed from the M. polymorpha model by up to 88 percent, particularly at the extremes of the data range examined, underestimating biomass in small trees and overestimating in large trees. The existing D. viscosa model underestimated biomass across all sizes by a mean of 43 percent compared to our model. The species‐specific models presented here should enable more accurate estimates of biomass and carbon sequestration in Hawaiian forests and shrublands.     

Pathway of Propionate Oxidation by a Syntrophic Culture of Smithella propionica and Methanospirillum hungatei

The pathway of propionate conversion in a syntrophic coculture of Smithella propionica and Methanospirillum hungatei JF1 was investigated by ¹³C-NMR spectroscopy. Cocultures produced acetate and butyrate from propionate. [3-¹³C]propionate was converted to [2-¹³C]acetate, with no [1-¹³C]acetate formed. Butyrate from [3-¹³C]propionate was labeled at the C2 and C4 positions in a ratio of about 1:1.5. Double-labeled propionate (2,3-¹³C) yielded not only double-labeled acetate but also single-labeled acetate at the C1 or C2 position. Most butyrate formed from [2,3-¹³C]propionate was also double labeled in either the C1 and C2 atoms or the C3 and C4 atoms in a ratio of about 1:1.5. Smaller amounts of single-labeled butyrate and other combinations were also produced. 1-¹³C-labeled propionate yielded both [1-¹³C]acetate and [2-¹³C]acetate. When ¹³C-labeled bicarbonate was present, label was not incorporated into acetate, propionate, or butyrate. In each of the incubations described above, ¹³C was never recovered in bicarbonate or methane. These results indicate that S. propionica does not degrade propionate via the methyl-malonyl-coenzyme A (CoA) pathway or any other of the known pathways, such as the acryloyl-CoA pathway or the reductive carboxylation pathway. Our results strongly suggest that propionate is dismutated to acetate and butyrate via a six-carbon intermediate.     

Differential gene expression analysis of strawberry cultivars that differ in fruit-firmness

Firmness is an important selection criterium in the breeding of fruit, including strawberry ( Fragaria  ×  ananassa Duch.). Clear differences in fruit-firmness are observed between cultivars. In order to identify candidate genes which might be associated with such textural differences, gene expression levels were compared for a soft and a firm cultivar (cv. Gorella and cv. Holiday, respectively). DNA-microarrays representing 1701 strawberry cDNAs were used for simultaneous hybridization of two RNA populations derived from red ripe fruit of both cultivars. In total 61 clones (3.6% of the total cDNAs on the arrays) displayed differential expression, including 10 clones (8 different ones) which showed homology to cell wall related genes in the public databases. The results from the microarray experiments were further confirmed by RNA gel blots, which were also used to examine gene expression in a third cultivar, Elsanta, showing an intermediate texture phenotype (offspring of a cross between Gorella and Holiday). Interestingly, two genes encoding proteins catalysing successive reactions in lignin metabolism (cinnamoyl CoA reductase and cinnamyl alcohol dehydrogenase) showed the highest difference in expression level.     

Long-range oxidative damage in duplex DNA: the effect of bulged G in a G–C tract and tandem G/A mispairs

Two series of duplex DNA oligomers were prepared having an anthraquinone derivative (AQ) covalently linked at a 5′-terminus. Irradiation of the AQ at 350 nm leads to injection of an electron hole (radical cation) into the DNA. The radical cation migrates through the DNA causing reaction primarily at G_n sequences. In one series, GA tandem mispairs are inserted between GG steps to assess the effect of the mispair on the transport of the radical cation, reaction (damage) caused by the radical cation at the mispair, and repair of the resulting damage by formamidopyrimidine DNA glycosylase (Fpg). In the second series, a bulged guanine in a G₃C₂ sequence is interposed between the GG steps. These experiments reveal that neither G/A tandem mispairs nor bulged guanines are significant barriers to long-range charge migration in DNA. The radical cation does not cause reaction at guanines in the G/A tandem mispair. Reaction does occur at the bulged guanine, but it is repaired by Fpg.     

Dynamics of Aboveground and Soil Carbon and Nitrogen Stocks and Cycling of Available Nitrogen along a Land-use Gradient in Rondônia,Brazil

High rates of deforestation in the Brazilian Amazon have the potential to alter the storage and cycling of carbon (C) and nitrogen (N) across this region. To investigate the impacts of deforestation, we quantified total aboveground biomass (TAGB), aboveground and soil pools of C and N, and soil N availability along a land-use gradient in Rondônia, Brazil, that included standing primary forest, slashed primary and secondary forest, shifting cultivation, and pasture sites. TAGB decreased substantially with increasing land use, ranging from 311 and 399 Mg ha^–1 (primary forests) to 63 Mg ha^–1 (pasture). Aboveground C and N pools declined in patterns and magnitudes similar to those of TAGB. Unlike aboveground pools, soil C and N concentrations and pools did not show consistent declines in response to land use. Instead, C and N concentrations were strongly related to percent clay content of soils. Concentrations of NO₃-N and NH₄-N generally increased in soils following slash-and-burn events along the land-use gradient and decreased with increasing land use. Increasing land use resulted in marked declines in NO₃-N pools relative to NH₄-N pools. Rates of net nitrification and N-mineralization were also generally higher in postfire treatments relative to prefire treatments along the land-use gradient and declined with increasing land use. Results demonstrate the linked responses of aboveground C and N pools and soil N availability to land use in the Brazilian Amazon; steady reductions in aboveground pools along the land-use gradient were accompanied by declines in inorganic soil N pools and transformation rates.