A first insight into population structure and linkage disequilibrium in the US peanut minicore collection

Knowledge of genetic diversity, population structure, and degree of linkage disequilibrium (LD) in target association mapping populations is of great importance and is a prerequisite for LD-based mapping. In the present study, 96 genotypes comprising 92 accessions of the US peanut minicore collection, a component line of the tetraploid variety Florunner, diploid progenitors A. duranensis (AA) and A. ipaënsis (BB), and synthetic amphidiploid accession TxAG-6 were investigated with 392 simple sequence repeat (SSR) marker bands amplified using 32 highly-polymorphic SSR primer pairs. Both distance- and model-based (Bayesian) cluster analysis revealed the presence of structured diversity. In general, the wild-species accessions and the synthetic amphidiploid grouped separately from most minicore accessions except for COC155, and were eliminated from most subsequent analyses. UPGMA analysis divided the population into four subgroups, two major subgroups representing subspecies fastigiata and hypogaea, a third group containing individuals from each subspecies or possibly of mixed ancestry, and a fourth group, either consisting of COC155 alone if wild species were excluded, or of COC155, the diploid species, and the synthetic amphidiploid. Model-based clustering identified four subgroups- one each for fastigiata and hypogaea subspecies, a third consisting of individuals of both subspecies or of mixed ancestry predominantly from Africa or Asia, and a fourth group, consisting of individuals predominantly of var fastigiata, peruviana, and aequatoriana accessions from South America, including COC155. Analysis of molecular variance (AMOVA) revealed statistically-significant (P < 0.0001) genetic variance of 16.87% among subgroups. A total of 4.85% of SSR marker pairs revealed significant LD (at r² ≥ 0.1). Of the syntenic marker pairs separated by distances < 10 cM, 11–20 cM, 21–50 cM, and > 50 cM, 19.33, 5.19, 6.25 and 5.29% of marker pairs were found in strong LD (P ≤ 0.01), in accord with LD extending to great distances in self pollinated crops. A threshold value of r² > 0.035 was found to distinguish mean r² values of linkage distance groups statistically from the mean r² values of unlinked markers; LD was found to extend to 10 cM over the entire minicore collection by this criterion. However, there were large differences in r² values among marker pairs even among tightly-linked markers. The implications of these findings with regard to the possibility of using association mapping for detection of genome-wide SSR marker-phenotype association are discussed.     

Pulsatile secretion of gonadotrophins, ovarian steroids and ovarian oxytocin during prostaglandin-induced regression of the corpus luteum in the cow

A luteolytic dose (500 micrograms) of cloprostenol was given on Day 12 of the oestrous cycle to 5 heifers. Blood samples were collected simultaneously from the caudal vena cava and jugular vein at 5-20-min intervals from -6 to 0 (control period), 0 to 12 and 24 to 36 h after PG injection. Pulses of LH were secreted concomitantly with pulses of FSH during all sampling periods. However, during the control period separate FSH pulses were detected resulting in a shorter (P less than 0.01) interpulse interval for FSH than LH (93 versus 248 min). LH and FSH pulse frequencies increased (P less than 0.01) beginning 1-3 h after PG to interpulse intervals of 59 and 63 min, respectively, and continued to be maintained 24-36 h after PG. Concomitantly there was a 2-3-fold increase (P less than 0.01) in basal concentrations and pulse amplitude for LH (but not FSH). FSH basal concentrations and pulse amplitudes decreased (P less than 0.05) in 3 heifers 24-36 h after PG. Pulsatile secretion of oestradiol was observed at frequencies similar to LH during the periods 4-12 h (3 heifers) and 24-36 h (2 heifers) after PG, respectively, resulting in higher (P less than 0.05) mean oestradiol concentrations. Progesterone concentrations in the vena cava increased (P less than 0.01) 5-10 min after PG but decreased (P less than 0.01) 67% by 20 min after PG. This decrease was followed by a rise (P less than 0.05) beginning 2-3 h after PG and lasting for an average of 3.3 h.(ABSTRACT TRUNCATED AT 400 WORDS)     

Experimental investigations of benthic reentry by migrating meiobenthic copepods

The resettlement behavior of meiobenthic copepods, which actively migrated from sediments in a seagrass bed, was investigated in a shallow subtidal area in Tampa Bay, Florida, U.S.A. Experimental studies were conducted to determine whether meiobenthic copepods after emerging from sediments at sunset reenter the sedimentary substratum or select other subhabitats, water and seagrass blades. Migrating copepods were collected with emergence traps and transferred to experimental aquaria in the field which contained sediment, seagrass-blade and water treatments. Settlement into each type of treatment was measured in separate 2-h and 9-h experiments. Differences in densities of copepod taxa retrieved from emergence traps and introduced into experimental aquaria were recorded as were differing relative proportions of each copepod species returning to the substratum treatments. Settlement patterns of total copepods and three dominant copepod species, Zausodes arenicolus, Halicyclops sp. and Robertsonia hamata, departed from those expected by chance. The populations of R. hamata and Halicyclops sp. which settled were generally skewed towards males and a close matching of males and copepodites within treatment dishes was evident. Similar to nighttime-emergence patterns, timing and magnitude of postmigration reentry differs among copepod taxa and such reentry may be linked to reproductive events. Complex behavioral processes previously noted for fish and macrofaunal organisms in seagrass beds may also be important in recruitment and reassortment of meiobenthic copepods.     

Persistence and growth of fecal Bacteroidales assessed by bromodeoxyuridine immunocapture

Extraintestinal growth of fecal bacteria can impair accurate assessment of watershed health. Anaerobic fecal bacteria belonging to the order Bacteroidales are attractive candidates for fecal source tracking because they have host-specific distributions and do not grow well in the presence of high oxygen concentrations. Growth of general and human-specific fecal Bacteroidales marker organisms in environmental samples (sewage) and persistence of the corresponding genetic markers were investigated using bromodeoxyuridine (BrdU) DNA labeling and immunocapture, followed by PCR detection. Background amplification of unlabeled controls occasionally occurred when a high number of PCR cycles was used. By using fluorescent detection of PCR products obtained after 15 cycles, which was determined to be quantitative, we enriched for BrdU-labeled DNA and did not detect unlabeled DNA. By using pure cultures of Bacteroides vulgatus, the ability of Bacteroidales bacteria to take up and incorporate BrdU into nascent DNA was confirmed. Fecal Bacteroidales organisms took up and incorporated BrdU into DNA during growth. In sewage incubated aerobically at the in situ temperature, Bacteroidales genetic marker sequences persisted for at least 24 h and Bacteroidales fecal bacteria grew for up to 24 h as well. Detection by PCR using a low, quantitative cycle number decreased the sensitivity of the assay such that we were unable to detect fecal Bacteroidales human-specific marker sequences in unlabeled or BrdU-labeled fractions, even when fluorescent detection was used. Using 30 PCR cycles with unlabeled fractions, human-specific Bacteroidales sequences were detected, and they persisted for up to 24 h in sewage. These data support the utility of BrdU labeling and immunocapture followed by length heterogeneity PCR or fluorescent detection using low numbers of PCR cycles. However, this method may not be sensitive enough to identify cells that are present at low densities in aquatic environments.     

Radiosensitivity of Mammalian Cells: IV. Effects of X-Irradiation on the DNA Synthetic Period in Synchronized Cells

The effect of X-irradiation on the timing of DNA synthesis in the Chinese hamster ovary cells has been investigated. Mitotically synchronized cells irradiated in mitosis or early G₁ exhibited a fixed, dose-independent (150-2000 rad) delay of 1.6 hr in entry into S, while the duration of S was unaffected. Cells irradiated during late G₁ or the first 0.8 hr of S were not affected either in time of initiation or duration of S. However, when cells 0.8 hr or more into S were irradiated, completion but not initiation of DNA synthesis was delayed, indicating a very precise separation of X-ray effects upon initiation and replication. There was no indication of a re-ordering of cells following irradiation and recovery, since cells in G₂ at the time of irradiation always divided before cells irradiated in S. The results suggest that two separate functions required for initiation and continued replication of DNA may be differentially sensitive to X-irradiation.     

The effect of polyamines on tyrosine hydroxylase and L-Aromatic amino acid decarboxylase

The polyamines stimulated tyrosine hydroxylase in whole homogenates of bovine caudate nuclei approximately 2 fold. TheV _max forl-tyrosine increased by 2.3 fold while theK _m s forl-tyrosine and for the cofactor (DMPH₄) were unchanged.l-Aromatic amino acid decarboxylase from whole rat brain homogenate was stimulated by about 40% in the presence of polyamines. These findings suggest that increased polyamine levels associated with increased cellular synthetic activity can modify the synthesis of neurotransmitters.     

Effect of alloxan-diabetes on multiple forms of hexokinase in adipose tissue and lung

Comparison has been made of the effect of alloxan-diabetes on the multiple forms of hexokinase (EC 2.7.1.1) in adipose tissue and lung. Types I and II hexokinase were distinguished in adipose tissue by their different stabilities to heat treatment, which made it possible to determine the activity of each form spectrophotometrically; additional confirmatory evidence was obtained from starch-gel electrophoresis. Type II hexokinase was markedly depressed in adipose tissue from alloxan-diabetic rats. Lung contained types I, II and III hexokinase, type I predominating. There was no significant change in the pattern of these multiple forms of hexokinase in lung from alloxan-diabetic rats. These results are discussed in relation to current ideas that the insulin-sensitivity of a tissue may be correlated with the content of type II hexokinase.     

Determinants of TRPV4 activity following selective activation by small molecule agonist GSK1016790A

TRPV4 (Transient Receptor Potential Vanilloid 4) channels are activated by a wide range of stimuli, including hypotonic stress, non-noxious heat and mechanical stress and some small molecule agonists (e.g. phorbol ester 4α-PDD). GSK1016790A (GSK101) is a recently discovered specific small molecule agonist of TRPV4. Its effects on physical determinants of TRPV4 activity were evaluated in HeLa cells transiently transfected with TRPV4 (HeLa-TRPV4). GSK101 (10 nM) causes a TRPV4 specific Ca(2+) influx in HeLa-TRPV4 cells, but not in control transfected cells, which can be inhibited by ruthenium red and Ca(2+)-free medium more significantly at the early stage of the activation rather than the late stage, reflecting apparent partial desensitization. Western blot analysis showed that GSK101 activation did not induce an increase in TRPV4 expression at the plasma membrane, but caused an immediate and sustained downregulation of TRPV4 on the plasma membrane in HeLa-TRPV4 cells. Patch clamp analysis also revealed an early partial desensitization of the channel which was Ca(2+)-independent. FRET analysis of TRPV4 subunit assembly demonstrated that the GSK101-induced TRPV4 channel activation/desensitization was not due to alterations in homotetrameric channel formation on the plasma membrane. It is concluded that GSK101 specifically activates TRPV4 channels, leading to a rapid partial desensitization and downregulation of the channel expression on the plasma membrane. TRPV4 subunit assembly appears to occur during trafficking from the ER/Golgi to the plasma membrane and is not altered by agonist stimulation.