A bicistronic transgene system for genetic modification of <Emphasis Type="Italic">Parthenium argentatum</Emphasis>

Parthenium argentatum (guayule) was transformed with a bicistronic transgene containing a viral 2A cleavage sequence. The transgene includes the coding sequences of two key enzymes of the mevalonate pathway, 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) and farnesyl pyrophosphate synthase (FPPS), involved in rubber biosynthesis. The viral 2A peptide sequence located between the two transgenes allowed for their co-expression via the Arabidopsis CBF2 (C-Binding repeat Factor 2) cold-inducible promoter. We identified three independent transgenic lines expressing the bicistronic transgenes upon cold treatment and examined the rubber content in the in vitro guayule plants.     

Life cycle inventory data

This article describes the work carried out by the Promoting Sound Practices (PSP) working group of SPOLD (Society for the Promotion of Lifecycle Development) on the development of a common format for reporting life cycle inventory data in a comparable and transparent way, and hence towards the eventual goal of a decentralised network of life cycle inventory databases. Establishing such a database network depends on the achievement of consensus amongst potential users, data owners and data generators. Accordingly, building consensus has been, and will continue to be given, a high priority in this work. As well as a summary of the consensus building activities, this article provides an outline of the developing format and an indication of the next steps planned, some of which are already underway.     

Participation of Auxin Transport in the Early Response of the <i>Arabidopsis</i> Root System to Inoculation with <i>Azospirillum brasilense</i>

The potential of Plant Growth Promoting Rhizobacteria (PGPR) has been demonstrated in the case of plant inoculation with bacteria of the genus Azospirillum which improves yield. A. brasilense produces a wide variety of molecules, including the natural auxin indole-3-acetic acid (IAA), as well as other phytoregulators. However, several studies have suggested that auxin induces changes in plant development during their interaction with the bacteria. The effects of A. brasilense Sp245 on the development of Arabidopsis thaliana root were investigated to help explain the molecular basis of the interaction. The results obtained showed a decrease in primary root length from the first day and remained so throughout the exposure, accompanied by a stimulation of initiation and maturation of lateral root primordia and an increase of lateral roots. An enhanced auxin response was evident in the vascular tissue and lateral root meristems of inoculated plants. However, after five days of bacterization, the response disappeared in the primary root meristems. The role of polar auxin transport (PAT) in auxins relocation involved the PGP1, AXR4-1, and BEN2 proteins, which apparently mediated A. brasilense-induced root branching of Arabidopsis seedlings.     

Phenotypic flexibility in basal metabolic rate is associated with rainfall variability among populations of rufous-collared sparrow

Phenotypic flexibility in metabolic rates allows organisms to reversibly adjust their energy flow to meet challenges imposed by a variable environment. In turn, the food habits hypothesis (FHH) predicts that species or populations adjust their basal metabolic rate (BMR) according to the diet attributes such as food abundance or predictability. Desert ecosystems represent a temporally heterogeneous environment because of low rain pulse predictability, which is also associated with temporal variation in food resources. In the present study, we investigated the relationship between the magnitude of BMR flexibility in response to dietary acclimation and the inter-annual rainfall variability in three populations of rufous-collared sparrows. Specifically we addressed the question of whether birds from a desert environment are more flexible in BMR than those from non-desert habitats. We found a positive trend between BMR flexibility and the inter-annual rainfall variability. In fact, dietary treatments had a significant effect only in desert birds, a result that also supported the FHH. Our study confirms the existence of phenotypic variation in response to environmental conditions among populations, and also highlights the importance of considering the circumstances in which phenotypic flexibility evolves and the specific environmental cues that induce their expression.     

Transcriptome responses to aluminum stress in roots of aspen (<Emphasis Type="Italic">Populus tremula</Emphasis>)

Background  

Ionic aluminum (mainly Al³⁺) is rhizotoxic and can be present in acid soils at concentrations high enough to inhibit root growth. Many forest tree species grow naturally in acid soils and often tolerate high concentrations of Al. Previously, we have shown that aspen (Populus tremula) releases citrate and oxalate from roots in response to Al exposure. To obtain further insights into the root responses of aspen to Al, we investigated root gene expression at Al conditions that inhibit root growth.     

A Single Amino Acid Difference Accounts for the Pharmacological Distinctions Between the Rat and Human 5-Hydroxytryptamine1B Receptors

Abstract: Molecular cloning of the rat and human 5-hydroxytryptamine_1B (5-HT_1B) receptors has revealed that the primary amino acid sequence of these two receptors is >90% identical. Despite this high degree of primary sequence homology, these two receptors have significantly different pharmacological properties. A mutant human 5-HT_1B receptor was constructed in which Thr³⁵⁵ was replaced by Asn, the corresponding residue at this position in the rat 5-HT_1B receptor. The pharmacology of the mutant human 5-HT_1B receptor was very similar to that of the rat 5-HT_1B receptor. Specifically, the mutant receptor had much higher affinity for pindolol, [¹²⁵I]-iodocyanopindolol, propranolol, and CP-93,129 than the wild-type receptor. In contrast, the mutant had significantly lower affinity for sumatriptan, N,N -dipropyl-5-carboxamidotryptamine, 5-methoxy- N,N -dimethyltryptamine, methysergide, metergoline, and rauwolscine. These data suggest that a single amino acid difference at position 355 is responsible for the pharmacological differences between the rat and human 5-HT_1B receptors.     

The Escherichia coli YadB gene product reveals a novel aminoacyl-tRNA synthetase like activity

In the course of a structural genomics program aiming at solving the structures of Escherichia coli open reading frame products of unknown function, we have determined the structure of YadB at 1.5A using molecular replacement. The YadB protein is 298 amino acid residues long and displays 34% sequence identity with E.coli glutamyl-tRNA synthetase (GluRS). It is much shorter than GluRS, which contains 468 residues, and lacks the complete domain interacting with the tRNA anticodon loop. As E.coli GluRS, YadB possesses a Zn2+ located in the putative tRNA acceptor stem-binding domain. The YadB cluster uses cysteine residues as the first three zinc ligands, but has a weaker tyrosine ligand at the fourth position. It shares with canonical amino acid RNA synthetases a major functional feature, namely activation of the amino acid (here glutamate). It differs, however, from GluRSs by the fact that the activation step is tRNA-independent and that it does not catalyze attachment of the activated glutamate to E.coli tRNAGlu, but to another, as yet unknown tRNA. These results suggest thus a novel function, distinct from that of GluRSs, for the yadB gene family.     

Density and population structure of the jaguar (<Emphasis Type="Italic">Panthera onca</Emphasis>) in a protected area of Los Llanos,Venezuela, from 1 year of camera trap monitoring

Density is crucial for understanding large carnivore ecology and conservation, but estimating it has proven methodologically difficult. We conducted 1 year of camera trapping to estimate jaguar (Panthera onca) density and population structure in the Los Llanos region of Venezuela on the Hato Piñero ranch, where hunting is prohibited and livestock are excluded from half of ranch lands. We identified 42 different jaguars and determined their sex, age class, and reproductive status. We estimated adult jaguar densities with spatial capture-recapture models, using sex/reproductive state and session as covariates. Models without temporal variation received more support than models that allowed variation between sessions. Males, reproductive females, and nonreproductive females differed in their density, baseline detectability, and movement. The best estimate of total adult jaguar population density was 4.44 individuals/100 km². Based on reproductive female density and mean number of offspring per female, we estimated cub density at 3.23 individuals/100 km² and an overall density of 7.67 jaguars/100 km². Estimated jaguar population structure was 21% males, 11% nonreproductive females, 26% reproductive females, and 42% cubs. We conclude that extending the sampling period to 1 year increases the detectability of females and cubs and makes density estimates more robust as compared to the more common short studies. Our results demonstrate that the Venezuelan Llanos represent important jaguar habitat, and further, they emphasize the importance of protected areas and hunting restrictions for carnivore conservation.     

Kinetochore Fiber Maturation in PtK1 Cells and Its Implications for the Mechanisms of Chromosome Congression and Anaphase Onset

Kinetochore microtubules (kMts) are a subset of spindle microtubules that bind directly to the kinetochore to form the kinetochore fiber (K-fiber). The K-fiber in turn interacts with the kinetochore to produce chromosome motion toward the attached spindle pole. We have examined K-fiber maturation in PtK₁ cells using same-cell video light microscopy/serial section EM. During congression, the kinetochore moving away from its spindle pole (i.e., the trailing kinetochore) and its leading, poleward moving sister both have variable numbers of kMts, but the trailing kinetochore always has at least twice as many kMts as the leading kinetochore. A comparison of Mt numbers on sister kinetochores of congressing chromosomes with their direction of motion, as well as distance from their associated spindle poles, reveals that the direction of motion is not determined by kMt number or total kMt length. The same result was observed for oscillating metaphase chromosomes. These data demonstrate that the tendency of a kinetochore to move poleward is not positively correlated with the kMt number. At late prometaphase, the average number of Mts on fully congressed kinetochores is 19.7 ± 6.7 (n = 94), at late metaphase 24.3 ± 4.9 (n = 62), and at early anaphase 27.8 ± 6.3 (n = 65). Differences between these distributions are statistically significant. The increased kMt number during early anaphase, relative to late metaphase, reflects the increased kMt stability at anaphase onset. Treatment of late metaphase cells with 1 μM taxol inhibits anaphase onset, but produces the same kMt distribution as in early anaphase: 28.7 ± 7.4 (n = 54). Thus, a full complement of kMts is not sufficient to induce anaphase onset. We also measured the time course for kMt acquisition and determined an initial rate of 1.9 kMts/min. This rate accelerates up to 10-fold during the course of K-fiber maturation, suggesting an increased concentration of Mt plus ends in the vicinity of the kinetochore at late metaphase and/or cooperativity for kMt acquisition.     

Intraspecific basal metabolic rate varies with trophic level in rufous-collared sparrows

One of the most controversial hypotheses that associate basal metabolic rate (BMR) with food habits and habitat productivity is the food habit hypothesis (FHH). Here we examined the relationship between BMR, diet, and climate among populations of the omnivorous passerine, Zonotrichia capensis (Emberizidae). We used nitrogen stable isotopes to estimate each individual's relative trophic level. To tease apart the effect of climatic variables and diet on BMR, we also used structural equation modeling. After the effect of body mass and climatic variables was taken into account, a significant effect of trophic level as estimated by δ¹⁵N on BMR was found. Our result seems to support the FHH at the intraspecific level, i.e., birds from the lower trophic levels – feeding on seeds and bud – had higher BMR than individuals from higher trophic levels.