Isolation of selective 3H-chlornaltrexamine-bound complexes,possible opioid receptor components in brains of mice

The nonequilibrium narcotic antagonist, chlornaltrexamine (CNA) was used to bind selectively and covalently pioid specific sites on brain membrane preparations. Selective binding of [³H]CNA occured with a saturation maximum of 185 fmol/mg protein. Bound [³H]CNA was extracted with Triton X-100, dialyzed against Brij 36T, precipitated with trichloroacetic acid and chromatographed on an ultrogel AcA 22 column. The elution profile suggests that this extract contains a minimum of four selective [³H]CNA complexes. At least two of these complexes migrate in a single large peak. Column calibration showed that this peak eluted at 590,000 daltons. One of these specific [³H]CNA complexes elutes at the elution volume of the column and is dialyzable. Finally, putative aggregate of these complexes elutes with the void volume.     

DEVELOPMENT AND ORGANIZATION OF THE SECONDARY VESSEL SYSTEM IN POPULUS GRANDIDENTATA

The apical 22 cm of a dormant, first-year sprout of Populus grandidentata was sectioned serially, and the primary and secondary xylem systems were studied microscopically and graphically reconstructed. A total of 15 nodes was present on the mature stem and 14 foliar primordia in the dormant bud. The vascular traces in the lower portion of the mature stem conformed to a 2/5 phyllotaxy while those of the upper portion and within the dormant bud conformed to a 3/8 phyllotaxy. The 2/5 to 3/8 phyllotactic transition occurred in an extremely precise and systematic two-step pattern: (1) The lateral traces shifted to a new point of origin on the parent central trace, and (2) three new central traces were initiated in sequence by divergences from left-traces. Metaxylem, when followed downward, conformed to the arrangement of the procambial trace system only within one orthostichy. Below this point, the metaxylem components of lateral traces physically separated from those of the protoxylem and continued downward on a new course. Metaxylem vessels produced by the trace cambium originated from a postulated vessel-generating center at the stem-petiole junction. Each metaxylem vessel developing basipetally through the primary body was continuous with a secondary vessel developing basipetally in the secondary body. Because secondary development closed the vascular cylinder, vessels originating from developing leaves or primordia situated at higher levels in the shoot were displaced radially outward when they entered the secondary xyelm. The distribution of vessels in the secondary xylem can therefore be accounted for by a knowledge of the production and distribution of metaxylem vessels in the primary body.     

Incorporation of C-photosynthate into major chemical fractions of source and sink leaves of cottonwood

The incorporation and distribution of photosynthetically fixed ¹⁴CO₂ was followed for 48 hours in a recently matured source leaf (LPI 7) and in young expanding source and sink leaves (LPI 4) of cottonwood (Populus deltoides Bartr.). The major chemical constituents of leaf laminae and petioles were separated by sequential solvent extractions and enzyme hydrolyses. Two hours after labeling, about 80% of the ¹⁴C was found in water-alcohol-soluble constituents in the mature source lamina as compared to about 45% in those of the young expanding leaf. In both mature and expanding source leaves the water-alcohol-soluble constituents decreased while the CHCl₃-soluble and -insoluble compounds increased with time. After 48 hours, 7 and 37% of the total ¹⁴C was recovered from structural carbohydrates and from protein + CHCl₃-soluble fractions, respectively, in the mature source leaf; and 4 and 65%, respectively, in the young source leaf. When the distribution of ¹⁴C among major chemical fractions was calculated on per cent dpm/mg basis, the data showed that a young sink leaf incorporated over twice as much ¹⁴C into structural carbohydrates as a young source leaf (11% versus 4%). However, when calculated on an absolute dpm/mg basis, activity in this fraction of the young source leaf exceeded that in the sink leaf by a ratio of about 11:1 (9528 versus 845 dpm/mg). Thus, most of the material for synthesis of structural carbohydrates was derived from in situ photosynthate.     

Correlations between net auxin and secondary xylem development in young Populus deltoides

Stems of cottonwood ( Populs deltoides Bartr. ex Marsh.) plants grown under different conditions were examined to determine the relation between net endogenous auxin yields and the acropetal advance of the primary-secondary vascular transition zone (TZ). In all treatments, the internode yielding maximum net auxin activity, as determined by the Avena curvature bioassay, closely corresponded with the internode in which the TZ occurred. Under short-day (SD) dormancy-inducing conditions, auxin yield declined steadily while the maximum auxin peak and the TZ shifted toward younger internodes. Auxin yields from these plants were extremely low after 5 weeks of SD compared with those from long-day (LD) plants. The only consistent auxin yield was obtained from internodes subtending young leaves beneath the apical bud. Plants placed in SD for 3 weeks and then returned to LD conditions showed an immediate increase in auxin yield in the stem, and the progressive acropetal advance of the TZ under SD was reversed. Therefore, within 7 LD the positions of the TZ and peak auxin yield corresponded to those observed before the imposition of SD Fully dormant plants placed in LD showed a dramatic rise in auxin yields during the first 2 weeks of renewed growth. Although low levels of auxin were found in the newly developing shoots after 6 LD, yields increased rapidly after 9 and 14 LD. The position of the TZ corresponded with the peak of net auxin activity after 9 and 14 LD.     

Attack of the PCR clones: Rates of clonality have little effect on RAD‐seq genotype calls

Interpretation of high‐throughput sequence data requires an understanding of how decisions made during bioinformatic data processing can influence results. One source of bias that is often cited is PCR clones (or PCR duplicates). PCR clones are common in restriction site‐associated sequencing (RAD‐seq) data sets, which are increasingly being used for molecular ecology. To determine the influence PCR clones and the bioinformatic handling of clones have on genotyping, we evaluate four RAD‐seq data sets. Data sets were compared before and after clones were removed to estimate the number of clones present in RAD‐seq data, quantify how often the presence of clones in a data set causes genotype calls to change compared to when clones were removed, investigate the mechanisms that lead to genotype call changes and test whether clones bias heterozygosity estimates. Our RAD‐seq data sets contained 30%–60% PCR clones, but 95% of RAD‐tags had five or fewer clones. Relatively few genotypes changed once clones were removed (5%–10%), and the vast majority of these changes (98%) were associated with genotypes switching from a called to no‐call state or vice versa. PCR clones had a larger influence on genotype calls in individuals with low read depth but appeared to influence genotype calls at all loci similarly. Removal of PCR clones reduced the number of called genotypes by 2% but had almost no influence on estimates of heterozygosity. As such, while steps should be taken to limit PCR clones during library preparation, PCR clones are likely not a substantial source of bias for most RAD‐seq studies.     

Leaf traits and performance vary with plant age and water availability in Artemisia californica

Background and AimsLeaf functional traits are strongly tied to growth strategies and ecological processes across species, but few efforts have linked intraspecific trait variation to performance across ontogenetic and environmental gradients. Plants are believed to shift towards more resource-conservative traits in stressful environments and as they age. However, uncertainty as to how intraspecific trait variation aligns with plant age and performance in the context of environmental variation may limit our ability to use traits to infer ecological processes at larger scales.MethodsWe measured leaf physiological and morphological traits, canopy volume and flowering effort for Artemisia californica (California sagebrush), a dominant shrub species in the coastal sage scrub community, under conditions of 50, 100 and 150 % ambient precipitation for 3 years.Key ResultsPlant age was a stronger driver of variation in traits and performance than water availability. Older plants demonstrated trait values consistent with a more conservative resource-use strategy, and trait values were less sensitive to drought. Several trait correlations were consistent across years and treatments; for example, plants with high photosynthetic rates tended to have high stomatal conductance, leaf nitrogen concentration and light-use efficiency. However, the trade-off between leaf construction and leaf nitrogen evident in older plants was absent for first-year plants. While few traits correlated with plant growth and flowering effort, we observed a positive correlation between leaf mass per area and performance in some groups of older plants.ConclusionsOverall, our results suggest that trait sensitivity to the environment is most visible during earlier stages of development, after which intraspecific trait variation and relationships may stabilize. While plant age plays a major role in intraspecific trait variation and sensitivity (and thus trait-based inferences), the direct influence of environment on growth and fecundity is just as critical to predicting plant performance in a changing environment.