Mapping genes essential for embryo development in Arabidopsis thaliana.

Summary We have previously isolated and characterized over 90 recessive mutants of Arabidopsis thaliana defective in embryo development. These emb mutants have been shown to differ in lethal phase, extent of abnormal development, and response in culture. We demonstrate in this report the value and efficiency of mapping emb genes relative to visible and molecular markers. Sixteen genes essential for embryo development were mapped relative to visible markers by analyzing progeny of selfed F₁ plants. Embryonic lethals are now the most common type of visible marker included on the linkage map of Arabidopsis. Backcrosses were used in several cases to orient genes relative to adjacent markers. Three genes were located to chromosome arms with telotrisomics by screening for a reduction in the percentage of aborted seeds produced by F₁ plants. A restriction fragment length polymorphism (RFLP) mapping strategy that utilizes pooled EMB/EMB F₂ plants was devised to increase the efficiency of mapping embryonic lethals relative to molecular markers. This strategy was tested by demonstrating that the biol locus of Arabidopsis is within 0.5 cM of an existing RFLP marker. Mapping embryonic lethals with both visible and molecular markers may therefore help to identify large numbers of genes with essential functions in Arabidopsis.     

Growth in vitro of arrested embryos from lethal mutants ofArabidopsis thaliana

Summary Seventeen embryo-lethal mutants ofArabidopsis thaliana with lethal phases ranging from the globular to mature cotyledon stages of development were analyzed by culturing arrested embryos on nutrient media designed to promote either callus formation or the completion of embryo development and the recovery of homozygous mutant plants. Enriched media supplemented with vitamins, amino acids, and nucleosides were used to identify potential auxotrophic mutants. Wild-type embryos produced extensive callus on basal and enriched media supplemented with 2,4-D and kinetin. Numerous roots developed when wildtype callus was grown in the presence of NAA and kinetin. Mutant embryos arrested prior to the heart stage of development formed only a slight amount of callus on basal and enriched media. Arrested embryos from mutants 122G-E and 112A-2A reached a later stage of development and gave the most interesting responses in culture. 122G-E mutant embryos failed to grow on basal media but produced extensive callus and homozygous mutant plants on enriched media. The specific nutrient required for growth of this mutant remains to be determined. Arrested embryos from mutant 112A-2A developed into abnormal plants without roots when placed in culture. Mutant callus also failed to form roots on a variety of root-inducing media. Expression of this mutant gene therefore disrupts development of the root apical meristem during both embryogenesis in vivo and organogenesis in vitro.     

Microbial biomass in a shallow, urban aquifer contaminated with aromatic hydrocarbons: analysis by phospholipid fatty acid content and composition

The city of Perth contains a number of sites that have been contaminated with hydrocarbons due to leakage from petroleum underground storage tanks. Microbial biomass in groundwater and sediment cores from above and below the water table, and from within and outside a plume of hydrocarbon contamination, was examined using phospholipid fatty acid methyl ester analysis. Microbial numbers, calculated from the phospholipid content, ranged from 0·9 × 10⁶ to 7·8 × 10⁶' Escherichia coli equivalent cells' g⁻¹ dry wt of sediment. Over 96% of the microbial biomass was attached to the sediment and the proportion of attached cells did not decrease within the plume of contaminants. The amount of biomass within aquifer samples seemed to be related more to the proximity of the rhizosphere to the shallow aquifer, and other unknown urban inputs, rather than to the effects of the plume of contaminants. Fatty acids common to many bacterial groups dominated within the plume, and as such the analyses gave limited insight into microbial community structure. For site assessment of intrinsic remediation of shallow aquifers in urban areas, estimates of microbial biomass may not provide information that is readily applicable to plume management.     

Attenuation of pattern recognition receptor signaling is mediated by a MAP kinase kinase kinase

Pattern recognition receptors (PRRs) play a key role in plant and animal innate immunity. PRR binding of their cognate ligand triggers a signaling network and activates an immune response. Activation of PRR signaling must be controlled prior to ligand binding to prevent spurious signaling and immune activation. Flagellin perception in Arabidopsis through FLAGELLIN‐SENSITIVE 2 (FLS2) induces the activation of mitogen‐activated protein kinases (MAPKs) and immunity. However, the precise molecular mechanism that connects activated FLS2 to downstream MAPK cascades remains unknown. Here, we report the identification of a differentially phosphorylated MAP kinase kinase kinase that also interacts with FLS2. Using targeted proteomics and functional analysis, we show that MKKK7 negatively regulates flagellin‐triggered signaling and basal immunity and this requires phosphorylation of MKKK7 on specific serine residues. MKKK7 attenuates MPK6 activity and defense gene expression. Moreover, MKKK7 suppresses the reactive oxygen species burst downstream of FLS2, suggesting that MKKK7‐mediated attenuation of FLS2 signaling occurs through direct modulation of the FLS2 complex.     

Influence of Observer Presence on Pacific Damsel Bug Behavior: Who is Watching Whom?

When direct observations are used to study animal behavior the presence of the observer may alter the animal and hence influence the data being collected, yet few studies have quantified this effect. We conducted direct observation studies in the glasshouse to assess the relative influence of observer presence on the behavior of Pacific damsel bugs, Nabis kinbergii, a potentially important predator of crop pests. Comparisons of predator activity, predator distribution, prey (Helicoverpa armigera) mortality and prey distribution between frequently observed and minimally observed treatments, during diurnal and nocturnal observation sessions showed that the frequency of observer presence had no apparent impact on Pacific damsel bug behavior. This is the first documented test of the impact of observer presence in an insect system. To place our results in context, we reviewed 15 papers on the influence of observer presence in a range of animals. We established that just over half of these papers found evidence for an effect. Nevertheless, direct observations should be useful in further studies of Pacific damsel bug behavior, and researchers using direct observations to study the behavior of other animals should be cognizant of observer effects during design and interpretation of their study.     

Novel Thermophilic Sulfate-Reducing Bacteria from a Geothermally Active Underground Mine in Japan

Thermophilic sulfate-reducing bacteria were enriched from samples obtained from a geothermal underground mine in Japan. The enrichment cultures contained bacteria affiliated with the genera Desulfotomaculum, Thermanaeromonas, Thermincola, Thermovenabulum, Moorella, “Natronoanaerobium,” and Clostridium. Two novel thermophilic sulfate-reducing strains, RL50JIII and RL80JIV, affiliated with the genera Desulfotomaculum and Thermanaeromonas, respectively, were isolated.     

Neuropsychopharmacology and Neurogenetic Aspects of Executive Functioning: Should Reward Gene Polymorphisms Constitute a Diagnostic Tool to Identify Individuals at Risk for Impaired Judgment?

Executive functions are processes that act in harmony to control behaviors necessary for maintaining focus and achieving outcomes. Executive dysfunction in neuropsychiatric disorders is attributed to structural or functional pathology of brain networks involving prefrontal cortex (PFC) and its connections with other brain regions. The PFC receives innervations from different neurons associated with a number of neurotransmitters, especially dopamine (DA). Here we review findings on the contribution of PFC DA to higher-order cognitive and emotional behaviors. We suggest that examination of multifactorial interactions of an individual's genetic history, along with environmental risk factors, can assist in the characterization of executive functioning for that individual. Based upon the results of genetic studies, we also propose genetic mapping as a probable diagnostic tool serving as a therapeutic adjunct for augmenting executive functioning capabilities. We conclude that preservation of the neurological underpinnings of executive functions requires the integrity of complex neural systems including the influence of specific genes and associated polymorphisms to provide adequate neurotransmission.     

The WSXWS motif in cytokine receptors is a molecular switch involved in receptor activation: insight from structures of the prolactin receptor

The prolactin receptor (PRLR) is activated by binding of prolactin in a 2:1 complex, but the activation mechanism is poorly understood. PRLR has?a conserved WSXWS motif generic to cytokine class I receptors. We have determined the nuclear magnetic resonance solution structure of the membrane proximal domain of the human PRLR and find that the tryptophans of the motif adopt a T-stack conformation in the unbound state. By contrast, in the hormone bound state, a Trp/Arg-ladder is formed. The conformational change is hormone-dependent and influences the receptor-receptor dimerization site 3. In the constitutively active, breast cancer-related receptor mutant PRLR(I146L), we observed a stabilization of the dimeric state and a change in the dynamics of the motif. Here we demonstrate a structural link between the WSXWS motif, hormone binding, and receptor dimerization and propose it?as?a general mechanism for class 1 receptor activation.