STS-PCR markers appropriate for wheat-barley introgression

Introgression of chromosomal segments across large taxonomic distances has long been an objective of scientists interested in understanding the relationships between genes and their effect on phenotype. Barley and wheat represent cultivated members of the Triticeae with different zones of adaptation, different responses to pathogens, and different end-use characteristics. Introduction of small, well-characterized chromosomal segments among grass relatives presents an opportunity to both better understand how genes perform in novel genomic environments and to learn more about the evolutionary novelties which differentiate related species. Since the distribution of the wheat-barley addition lines, the potential power and value of a comprehensive series of wheat/barley translocation lines has been widely appreciated. A scarcity of easy-touse markers which unambiguously distinguish barley loci from their wheat homologues has limited the ability of scientists to identify the relatively rare inter-chromosomal recombination events which are the necessary antecedents of these lines. Since the single most critical pathogen affecting U.S. wheat producers is Karnal bunt (Tilletia indica) and since barley carries a gene conferring immunity, molecular markers may prove practically and immediately important. In this report we describe a series of 135 barley-specific markers amplified by 115 primer sets developed from sequences from previously mapped restriction fragment length polymorphism (RFLP) markers. These easily distinguish the cognate barley products from their wheat counterparts and should find ready use in the identification of lines which contain wheat/barley translocation events.     

Differential Agonist Regulation of the Human κ-Opioid Receptor

Abstract: Opiates are potent analgesics used clinically in the treatment of pain. A significant drawback to the chronic use and clinical effectiveness of opiates is the development of tolerance. To investigate the cellular mechanisms of tolerance, the cloned human κ-opioid receptor was stably expressed in human embryonic kidney (HEK 293) cells, and the effects of opioid agonist treatment were examined. The receptor-expressing cells showed specific high-affinity membrane binding for a κ-selective opioid, ³H-labeled (+)-(5α,7α,8β)-N-methyl-N-[7-(1-pyrrolidinyl)-1-oxaspiro[4,5]dec-8-yl]benzeneacetamide ([³H]U69,593), and a nonselective opioid antagonist, [³H]diprenorphine. Pretreatment with pertussis toxin or guanosine 5′-O-(3-thiotriphosphate) reduced [³H]69,593 binding, indicating that the human κ receptor coupled to G proteins of the G_i or G_o families in HEK 293 cells. The receptor-mediated inhibition of adenylyl cyclase was abolished by pertussis toxin pretreatment and was blocked by a κ-selective antagonist, norbinal-torphimine. A 3-h pretreatment with a κ-selective agonist, (±)-trans-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)cyclohexyl]benzeneacetamide (U50,488), caused receptor down-regulation, whereas no receptor down-regulation was found after levorphanol pretreatment. U50,488 or dynorphin A_1–17 pretreatments (3 h) desensitized the ability of U50,488 or dynorphin A_1–17 to inhibit cyclic AMP accumulation, as evidenced by a decrease in functional potency. Also, U50,488 pretreatment desensitized the ability of levorphanol to inhibit forskolin-stimulated cyclic AMP accumulation. In contrast, pretreatment of cells with either levorphanol or a potent nonselective opioid, etorphine, resulted in no apparent receptor desensitization. Taken together, these results demonstrate that the human κ receptor is differentially regulated by selective and nonselective opioid agonists, with selective agonists able to desensitize the receptor.     

The preparation, characterisation and low temperature X-ray structure of Ru6(η-μ4-CO)2(CO)13(η6-C6Me6)

The new organometallic cluster (η²-μ₄-CO)₂(CO)₁₃(η⁶-C₆Me₆) has been prepared by the thermolysis of Ru₃(CO)₁₂ with hexamethylbenzene in octane and characterised by a single crystal X-ray diffraction study. It is isostructural with the known cluster Ru₆(η²-μ₄-CO)₂(CO)₁₃(η⁶-C₆H₃Me₃) and the metal core constitutnts the same tetrahedral Ru₄ unit with two edge-bridging Ru atoms. The mesitylene derivative has been shown to undergo rearrangement to afford the octahedral carbido cluster Ru₆C(CO)₁₄(η⁶-C₆H₃Me₃), but this conversion is not observed for the new hexamethylbenzene derivative.     

Turning radius of yellowfin tuna (Thunnus albacares) in unsteady swimming manoeuvres

The specific turning radius of yellowfin tuna Thunnus albacares Cuvier is large relative to that of other fish (0.47 total body length ± 0.18, mean ± 2 s.e .). We argue that specializations for efficient steady swimming compromise performance in transient turning manoeuvres.     

Linkage mapping of two mutations that reduce phytic acid content of barley grain

 This study describes the inheritance and linkage map positions of two low phytic acid barley (Hordeum vulgare) mutations, lpa1-1 and lpa2-1, that dramatically reduce grain phytic acid content and increase inorganic seed phosphorus (P). Wide-cross, F₂ mapping populations were constructed by mating six-rowed varieties, ‘Steptoe’ and/or ‘Morex’, with two-rowed ‘Harrington’lpa donor lines homozygous for either lpa1-1 or lpa2-1. The barley lpa1-1 mutation showed normal inheritance patterns, whereas a deficiency of homozygous lpa2-1/lpa2-1 F₂ plants was observed. We identified a codominant, STS-PCR marker (aMSU21) that cosegregated with lpa1-1 in a population of 41 F₂ plants. The aMSU21 marker was then mapped to a locus on barley chromosome 2H, using a North American Barley Genome Mapping Project (NABGMP) doubled haploid population (‘Harrington’בMorex’). We determined that lpa2-1 is located within a recombination interval of approximately 30 cM between two AFLP markers that were subsequently mapped to barley chromosome 7H by integration with the same NABGMP population. Recent comparative mapping studies indicate conserved genetic map orders of several homologous molecular marker loci in maize and the Triticeae species that also show corresponding linkage to the biochemically similar lpa2 mutations of maize and barley. This observation suggests that barley and maize lpa2 mutations may affect orthologous genes. No such evidence for correspondence of the phenotypically similar lpa1 mutations of barley and maize has been revealed. Received: 22 September 1997 / Accepted: 2 December 1997     

Abscisic acid induced electrolyte leakage in woody species with contrasting ecological requirements

To determine the relationship between drought, abscisic acid (ABA) levels, and membrane leakage, these aspects were studied in three woody species with contrasting ecological requirements. Physiological comparisons were made, during a 14-day drought-recovery cycle, on a drought resistant species, jack pine ( Pinus banksiana Lamb.), and two more drought susceptible species, black spruce ( Picea mariana (Mill) B.S.P.) and flooded gum ( Eucalyptus grandis W. Hill ex Maiden). As the xylem pressure potentials (Ψ), declined under drought, both foliar ABA and electrolyte leakage increased significantly. However, ABA increased prior to the increase in leakage, and there was a significant linear correlation between endogenous ABA and leakage in all 3 species. Addition of exogenous ABA to the bathing solutions also significantly increased leakage of both unstressed and (sorbitol) osmotically-stressed leaf segments in response to increasing (log) concentrations of (±) cis-trans ABA [0–10.0 μ M ]. Electrolyte leakage in black spruce and flooded gum was more sensitive to stress-induced ABA than in jack pine. The results suggest that drought increases foliar ABA which initiates membrane leakage, and that sensitivity of membranes to ABA accumulation varies, depending on the ecological requirements of a species.     

A study and model of the role of the Renshaw cell in regulating the transient firing rate of the motoneuron

 This study sought to investigate the role of the Renshaw cell with respect to transient motoneuron firing. By studying the cat motoneuron and Renshaw cell, several low-order lumped parameter models were developed that simulate the known characteristics of the injected input current vs. firing rate. The neuron models in the Renshaw cell inhibition configuration were tuned to fit experimental data from cat motoneurons. Models included both linear versions and those with sigmoidal nonlinearities. Results of the simulation indicate that the motoneuron itself provides the adaptation seen in its firing rate and that the Renshaw cell’s role is primarily to fine-tune the motoneuron’s adaptation process. Received: 23 July 1993/Accepted in revised form: 9 February 1994     

Identification of gibberellins in leaf tissues of strawberry (Fragaria × ananassa Duch.) grown under different photoperiods

In studies of the effect of long or short-day photoperiod treatments on the qualitative gibberellin (GA) content of mature leaves of a facultative short-day (SD) strawberry cultivar (Fragaria × ananassa Duch. cv. Elsanta), GA₁, GA₈, GA₁₇, GA₁₉, GA₂₀, GA₂₉ and GA₄₄ were identified by full-scan gas chromatography - mass spectrometry (GC-MS) in extracts from plants grown under long-day (LD) conditions, and GA₁, GA₅, GA₈, GA₁₉, GA₂₀ and GA₂₉ in similar extracts from plants subjected to eight SD cycles after growth under LD conditions. The early 13-hydroxylation GA biosynthetic pathway thus appeared to predominate, with the apparent absence of GA₅ in LD and of GA₁₇ and GA₄₄ in SD extracts providing evidence of modulation of this pathway by photoperiod. A search, including GC-MS with selected ion monitoring, failed to detect GA₃, or the polyhydroxylated GA₈₅, GA₈₆, GA₈₇ or GA₃₂ for which some extracts were specifically purified.This paper is respectfully dedicated to the memory of Gordon Browning, who died suddenly on the 1st July, 1993. He will be sorely missed, both as a friend and colleague.     

The emerging family of dystrophin-related proteins

Duchenne and Becker muscular dystrophies are caused by mutations in the gene encoding dystrophin, a component of the subsarcolemmal cytoskeleton. Dystrophin-related proteins are identical or homologous to the cysteine-rich and C-terminal domains of dystrophin. This part of dystrophin binds to a membrane-spanning glycoprotein complex in muscle. At least five dystrophin-related proteins are encoded by the Duchenne muscular dystrophy locus. These proteins are found in many non-muscle tissues where dystrophin is not expressed and they are thought to be membrane-associated. Two other dystrophin-related proteins--utrophin and an 87 kDa postsynaptic protein--are encoded by separate loci and, like dystrophin, they are components of the neuromuscular junction.