Direct Complementation ofChlamydomonasMutants with Amplified YAC DNA

We previously described the construction and characterization of aChlamydomonasgenomic library in yeast artificial chromosomes (YACs). Here we describe the isolation and genetic mapping of YACs at the FLA10 locus on theunichromosome as well as isolation of a YAC spanning the PF14 locus on chromosome VI. Genetic mapping of YAC end clones by RFLP analyses in interspecific crosses reveals that YACs with a physical size of 150 kb commonly span genetic intervals defined by one or two recombination events in crosses of approximately 20 tetrads. This promises to make chromosomal walking inChlamydomonasa relatively efficient enterprise. We also describe our development of a method for direct complementation of mutant genes by transformation with amplified wildtype YAC DNA. The use of positional cloning using YACs and this direct functional assay for the presence of a gene in a YAC represent powerful molecular genetic tools enabling the cloning of most anyChlamydomonasgene.     

A trait‐based approach for predicting species responses to environmental change from sparse data: how well might terrestrial mammals track climate change?

Estimating population spread rates across multiple species is vital for projecting biodiversity responses to climate change. A major challenge is to parameterise spread models for many species. We introduce an approach that addresses this challenge, coupling a trait‐based analysis with spatial population modelling to project spread rates for 15 000 virtual mammals with life histories that reflect those seen in the real world. Covariances among life‐history traits are estimated from an extensive terrestrial mammal data set using Bayesian inference. We elucidate the relative roles of different life‐history traits in driving modelled spread rates, demonstrating that any one alone will be a poor predictor. We also estimate that around 30% of mammal species have potential spread rates slower than the global mean velocity of climate change. This novel trait‐space‐demographic modelling approach has broad applicability for tackling many key ecological questions for which we have the models but are hindered by data availability.     

Simulation of forest tree species’ bud burst dates for different climate scenarios: chilling requirements and photo-period may limit bud burst advancement

This study investigates whether the assumed increase of winter and spring temperatures is depicted by phenological models in correspondingly earlier bud burst (BB) dates. Some studies assume that rising temperatures lead to an earlier BB, but even later BB has been detected. The phenological model PIM (promoter-inhibitor-model) fitted to the extensive phenological database of the German Weather Service was driven by several climate scenarios. This model accounts for the complicated mechanistic interactions between chilling requirements, temperature and photo-period. It predicts BB with a r² between 0.41 and 0.62 and a RMSE of around 1 week, depending on species. Parameter sensitivities depict species dependent interactions between growth and chilling requirements as well as photo-period. A mean trend to earlier BB was revealed for the period 2002– 2100, varying between ?0.05 and ?0.11 days per year, depending on species. These trends are lower than for the period 1951– 2009. Within climate scenario period, trends are decreasing for beech and chestnut, stagnating for birch and increasing for oak. Results suggest that not fulfilled chilling requirements accompanied by an increasing dependency on photo-period potentially limit future BB advancement. The combination of a powerful phenological model, a large scale phenological database and several climate scenarios, offers new insights into the mechanistic comprehension of spring phenology.     

Isolation and Functional Characterisation of a fads2 in Rainbow Trout (Oncorhynchus mykiss) with Δ5 Desaturase Activity

Rainbow trout, Oncorhynchus mykiss, are intensively cultured globally. Understanding their requirement for long-chain polyunsaturated fatty acids (LC-PUFA) and the biochemistry of the enzymes and biosynthetic pathways required for fatty acid synthesis is important and highly relevant in current aquaculture. Most gnathostome vertebrates have two fatty acid desaturase (fads) genes with known functions in LC-PUFA biosynthesis and termed fads1 and fads2. However, teleost fish have exclusively fads2 genes. In rainbow trout, a fads2 cDNA had been previously cloned and found to encode an enzyme with Δ6 desaturase activity. In the present study, a second fads2 cDNA was cloned from the liver of rainbow trout and termed fads2b. The full-length mRNA contained 1578 nucleotides with an open reading frame of 1365 nucleotides that encoded a 454 amino acid protein with a predicted molecular weight of 52.48 kDa. The predicted Fads2b protein had the characteristic traits of the microsomal Fads family, including an N-terminal cytochrome b5 domain containing the heme-binding motif (HPPG), histidine boxes (HDXGH, HFQHH and QIEHH) and three transmembrane regions. The fads2b was expressed predominantly in the brain, liver, intestine and pyloric caeca. Expression of the fasd2b in yeast generated a protein that was found to specifically convert eicosatetraenoic acid (20:4n-3) to eicosapentaenoic acid (20:5n-3), and therefore functioned as a Δ5 desaturase. Therefore, rainbow trout have two fads2 genes that encode proteins with Δ5 and Δ6 desaturase activities, respectively, which enable this species to perform all the desaturation steps required for the biosynthesis of LC-PUFA from C₁₈ precursors.     

Specific Laboratory Test for Diagnosis of Multiple Sclerosis

Lymphocytes from patients with multiple sclerosis are much more susceptible to the inhibitory activity of linoleic acid (0·08 mg/ml) when tested for sensitization to thyroid by the macrophage electrophoretic mobility test (91% inhibition) than are those from normal subjects (57% inhibition). Cells from patients with a variety of other neurological diseases give 47% inhibition with linoleic acid. These differences are specific for multiple sclerosis and can be used as an in-vitro diagnostic test for the disease. Nearly 43% of clinically normal near relatives of patients with multiple sclerosis show an “anomalous” figure of about 77%; in the remainder the figure is the same as in the general population (57%). An anomalous result is compatible with lifelong freedom from M.S. Possibly a congenital anomalous handling of unsaturated fatty acids is a constant feature of the disease.     

Material transfer from water to sediment in Florida lakes

We describe statistical relationships between chemical aspects of surficial sediments from 34 Florida lakes, and trophic state of the overlying waters. Trophic state is expressed by Carlson's TSI-Chl a. The objectives of the study are two-fold: 1) to understand processes that govern the transfer of material across the mudwater interface, and 2) to develop transfer functions for inferring historical TSI measures in chemically analyzed, ²¹⁰Pb-dated cores. Simple regression of organic matter content or nutrient (C, N, P) concentration in surface sediments vs. TSI yields nonsignificant or weak positive correlations. However, using a novel application of ²¹⁰Pb assay, net accumulation rates of the materials are estimated and show a better correlation with trophic state. Cation (Ca, Mg, Fe, K) and sulfur concentrations in surface muds are poorly related to corresponding TSI's. Net accumulation rates of these elements (Ca, Mg, Fe, K, S) are positively correlated (P < 0.01) with TSI-Chl a. Chemical data from the Florida surficial sediment survey suggest that inferred levels of past trophic state should be based on net accumulation rates of chemical constituents rather than their simple sediment concentrations.     

Intercoil connections of the kidney of the brine shrimp,Artemia salina

Summary Maxillary glands (kidneys) of adult brine shrimp were examined by light and electron microscopy. Within a single maxillary gland there are numerous places where adjacent coils of the efferent tubule are linked by structures designated intercoil connections. At the site of such a connection the basal laminae of the two coils are continuous, and one or more cytoplasmic processes originating from a cell of one coil form a bridge to the adjacent coil. The connecting processes are branched and interdigitate complexly with complementary processes from cells of the neighboring coil. Speculations are made about the functional significance of intercoil connections in the maxillary gland of Artemia.     

ULTRASTRUCTURE AND BIREFRINGENCE OF THE ISOLATED MITOTIC APPARATUS OF MARINE EGGS

Isolated mitotic apparatuses (MA) of clam and sea urchin eggs were investigated by polarizing and electron microscopy. Examination of fixed MA in oils of different refractive index revealed that at least 90% of the retardation of isolated MA is due to positive, form birefringence, the remaining retardation deriving from positive, intrinsic birefringence. Electron micrographs reveal the isolated MA to be composed of microtubules, ribosome-like particles, and a variety of vesicles. In the clam MA the number of vesicles and ribosome-like particles relative to the number of microtubules is much lower than in the sea urchin MA. In clam MA this allows form and intrinsic birefringence to be related directly to microtubules. The relation of birefringence to microtubules in isolated sea urchin MA is more complex since ribosome-like particles adhere to microtubules, are oriented by them, and are likely to contribute to the form birefringence of the isolated MA. However, comparison of values of retardation for clam and sea urchin MA, indicates that the major part of the birefringence in sea urchin MA is also due to microtubules. The interpretation of the structures giving rise to birefringence in the MA of the living cells is likely to be even more complex since masking substances, compression, or tension on the living MA may alter the magnitude or sign of the birefringence.     

The fine structure of the maxillary gland of the brine shrimp,Artemia salina: The efferent duct

Summary The efferent duct of the maxillary gland of adult brine shrimp, Artemia salina, is specialized into two morphologically distinct regions: an efferent tubule and a terminal duct. The wall of the efferent tubule is composed of epithelial cells which possess an apical microvillous border and, more basally, membranous configurations with which large numbers of mitochondria are closely associated. These membranous configurations are of two types: 1) infoldings of the plasma membrane of a single cell, and 2) interdigitations of lateral processes from adjoining cells. In contrast to the efferent tubule, the cells of the terminal duct possess a secreted cuticle and lack modifications which markedly increase the area of the plasma membrane. Mitochondria of the terminal duct are smaller and less numerous than those of the efferent tubule and typically are not found in close association with the plasma membrane. The ultrastructure of the efferent tubule and terminal duct suggests that the former region plays an active role in modifying the luminal contents and the latter region functions primarily as a conduit for the final urine.