ABSTRACT: RNA was extracted from xylem tissue of the first 5cm of the lower stem of four replicates of three overexpression lines and wild-type. Samples were collected from 3-month-old Populus from growth-chamber experiments at 25oC and 35oC. 2 treatments (growth at 25C and 35C temperature), 4 genotypes (wildtype and 3 35S::EVE1 independent lines) and 4 biological reps per genotype
Project description:RNA was extracted from xylem tissue of the first 5cm of the lower stem of four replicates of three overexpression lines and wild-type. Samples were collected from 3-month-old Populus from growth-chamber experiments at 25oC and 35oC. 2 treatments (growth at 25C and 35C temperature), 4 genotypes (wildtype and 3 35S::EVE1 independent lines) and 4 biological reps per genotype
Project description:In this study, we examined the transcriptomic responses to temperature acclimation (14oC, 20oC, and 25oC) in atrial and ventricular tissues of Pacific bluefin tuna (PBFT). A global gene expression analysis using a bluefin tuna-specific microarray indicated profound changes in expression of genes associated with energy metabolism, protein turnover, cellular stress response, oxidative stress and apoptosis. A principal component analysis revealed tissue-specific transcriptomic responses to temperature, with atrium at 25oC showing the greatest variation. Overall transcriptomic data suggests that PBFT can optimize cardiac function in the cold by acclimating to 14oC. Capacity to acclimate to colder temperatures potentially underlies this species ability to expand its vertical and horizontal thermal niche and migrate to colder oceans at high latitudes. In contrast, the cardiac phenotype of 25oC acclimated fish infers that PBFT hearts struggle to maintain cellular homeostasis and are subjected to programmed cell death. The goal of this study was to compare transcriptomic response to cold and warm temperature acclimations across cardiac tissues in Pacific bluefin tuna. Fish (n=4) were acclimated to 14C, 20C and 25C, and RNA was extracted from atrial, ventricular compact and spongy tissues. Experimental samples were hybridyzed against a reference pool that contained a mix of RNA from every sample.
Project description:Potato seedlings were subjected to cold, heat and salt stress. Expression profiles were captured at three different time-points, 3h, 9h and 27h from two different tissues, roots and leaves. The experiment was preformed independently three times. Commercially available true potato seeds (Variety Gilroy) were germinated on rafts floating on hydroponic medium in Magenta boxes. Plants were grown for 5 weeks prior to stress application under long day conditions (16h light and 8h dark) at 25C with gentle agitation. To initiate stress the medium was replaced with fresh medium pre-chilled to 4C (cold stress), pre-heated to 35C (heat stress) or supplemented with 100mM NaCl (salt stress). Cold and heat stress were maintained for the duration of the experiment by placing the Magenta boxes on ice or in a water-bath at 35C. For every individual sample two boxes of plants were used pooling a total of 6 plants per sample. For each time-point a single control sample was used by changing the media in a similar way as for the stress induction. A total of six boxes were combined for the pooled reference samples. Plants were harvested at the appropriate time and snap-frozen in liquid nitrogen. Roots and aerial tissue was separated prior to freezing. The tissue was stored at -80C freezer until isolation. Total RNA was isolated using RNeasy isolation kit. The intactness of the RNA was verified on gel and the concentration was adjusted to 3ug/ul by ethanol precipitation and re-suspension. Series_weblink: http://www.tigr.org/tdb/potato Keywords = potato, Abiotic stress
Project description:The experiment was conducted using four 11-year-old potted grapevines of V. vinifera L. cv. Cabernet Sauvignon grafted on SO4 (Selection Oppenheim No. 4) grown in a phytotron. Vines were trained on a Guyot trellising system and each vine carried 4-10 clusters of grapes. The experiment started approximately 1 week before veraison, when berry softening started, and continued to fruit maturity. The two temperature regimes consisted of a high day (06.00–20.00 h) temperature (max.35C) and a control (max. 25C). Under both conditions, the night-time (20.00–06.00 h) temperature was 20C. PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Kentaro Mori. The equivalent experiment is VV9 at PLEXdb developmental stage: 2 weeks - temperature: High (2-replications); developmental stage: 2 weeks - temperature: Control(2-replications); developmental stage: 4 weeks - temperature: High (2-replications); developmental stage: 4 weeks - temperature: Control(2-replications); developmental stage: 6 weeks - temperature: High (2-replications); developmental stage: 6 weeks - temperature: Control(2-replications)
Project description:Novel purine and purine isosteres containing a ferrocene motif and 4,1-disubstituted (<b>11a</b>-<b>11c</b>, <b>12a</b>-<b>12c</b>, <b>13a</b>-<b>13c</b>, <b>14a</b>-<b>14c</b>, <b>15a</b>-<b>15c</b>, <b>16a</b>, <b>23a</b>-<b>23c</b>, <b>24a</b>-<b>24c</b>, <b>25a</b>-<b>25c</b>) and 1,4-disubstituted (<b>34a</b>-<b>34c</b> and <b>35a</b>-<b>35c</b>) 1,2,3-triazole rings were synthesized. The most potent cytotoxic effect on colorectal adenocarcinoma (SW620) was exerted by the 6-chloro-7-deazapurine <b>11c</b> (IC<sub>50</sub> = 9.07 µM), 6-chloropurine <b>13a</b> (IC<sub>50</sub> = 14.38 µM) and <b>15b</b> (IC<sub>50</sub> = 15.50 µM) ferrocenylalkyl derivatives. The <i>N</i>-9 isomer of 6-chloropurine <b>13a</b> containing ferrocenylmethylene unit showed a favourable in vitro physicochemical and ADME properties including high solubility, moderate permeability and good metabolic stability in human liver microsomes.
Project description:In vertebrates, Evx homeodomain transcription factor-encoding genes are expressed in the posterior region during embryonic development, and overexpression experiments have revealed roles in tail development in fish and frogs. We analyzed the molecular mechanisms of posterior neural development and axis formation regulated by eve1. We show that eve1 is involved in establishing trunk and tail neural ectoderm by two independent mechanisms: First, eve1 posteriorizes neural ectoderm via induction of aldh1a2, which encodes an enzyme that synthesizes retinoic acid; second, eve1 is involved in neural induction in the posterior ectoderm by attenuating BMP expression. Further, eve1 can restore trunk neural tube formation in the organizer-deficient ichabod(-/-) mutant. We conclude that eve1 is crucial for the organization of the antero-posterior and dorso-ventral axis in the gastrula ectoderm and also has trunk- and tail-promoting activity.
Project description:In Arabidopsis, inflorescence stem formation is a critical process in phase transition from the vegetative to the reproductive state. Although inflorescence stem development has been reported to depend on the expression of a variety of genes during floral induction and repression, little is known about the molecular mechanisms involved in the control of inflorescence stem formation. By activation T-DNA tagging mutagenesis of Arabidopsis, a dominant gain-of-function mutation, eve1-D (eternally vegetative phase1-Dominant), which has lost the ability to form an inflorescence stem, was isolated. The eve1-D mutation exhibited a dome-shaped primary shoot apical meristem (SAM) in the early vegetative stage, similar to that seen in the wild-type SAM. However, the SAM in the eve1-D mutation failed to transition into an inflorescence meristem (IM) and eventually reached senescence without ever leaving the vegetative phase. The eve1-D mutation also displayed pleiotropic phenotypes, including lobed and wavy rosette leaves, short petioles, and an increased number of rosette leaves. Genetic analysis indicated that the genomic location of the EVE1 gene in Arabidopsis thaliana corresponded to a bacterial artificial chromosome (BAC) F4C21 from chromosome IV at ?17cM which encoded a novel ubiquitin family protein (At4g03350), consisting of a single exon. The EVE1 protein is composed of 263 amino acids, contains a 52 amino acid ubiquitin domain, and has no glycine residue related to ubiquitin activity at the C-terminus. The eve1-D mutation provides a way to study the regulatory mechanisms that control phase transition from the vegetative to the reproductive state.
Project description:We take one-year-old plants for high-temperature treatments and controls. We used the Affymetrix Poplar GeneChip to decrypt the gene functions and mechanisms in Populus simonii leaves. We used microarrays to detail the global program of gene expression during high-temperature treatments. Populus simonii leaves were taken from high-temperature (42C) treatments and controls (25C) for RNA extraction and hybridization on Affymetrix microarrays. H1 and H2 are from high-temperature treatments, CK1 and CK2 are controls.
Project description:Different intensities of high temperatures affect the growth of photosynthetic cells in nature. To elucidate the underlying mechanisms, we cultivated the unicellular green alga Chlamydomonas reinhardtii under highly controlled photobioreactor conditions and revealed systems-wide shared and unique responses to 24-hour moderate (35oC) and acute (40oC) high temperatures and subsequent recovery at 25oC. We identified transcripts/proteins with unique differential regulation at 35oC, uncovering previously overlooked novel elements in response to moderate high temperature. Heat at 35oC increased transcripts/proteins involved in gluconeogensis/glyoxylate-cycle for carbon uptake, promoted growth, and increased starch accumulation. Heat at 40oC inhibited growth, resulting in carbon uptake over usage and increased starch accumulation. Heat at 35oC transiently arrested the cell cycle followed by partial synchronization while 40oC inhibited DNA replication and arrested the cell cycle. Both high temperatures induced photoprotection, while 40oC decreased photosynthetic efficiencies, distorted thylakoid/pyrenoid ultrastructure, and affected the carbon concentrating mechanism. We demonstrated increased transcript/protein correlation during heat, which decreased during recovery, suggesting reduced post-transcriptional regulation during heat may help coordinate heat tolerance activities efficiently. During recovery after both treatments, transcripts/proteins related to DNA synthesis increased while those involved in photosynthetic light reactions decreased. We propose down-regulating photosynthetic light reactions during DNA replication benefits cell cycle resumption by reducing ROS production. Our results provide potential targets to increase thermotolerance in algae and crops. Overall design: Transcriptomic time-course analysis of wild-type Chlamydomonas reinhardtii cells exposed to 24-hours of moderate (35C) or acute (40C) high temperature, followed by a 48-hour recovery at 25C.
Project description:Streptococcus pneumoniae expresses capsular polysaccharides (CPSs) to protect itself from opsonophagocytic killing. The genes responsible for capsules synthesized by the Wzy-dependent mechanism, which accounts for 96 of the 98 known pneumococcal capsule types, are in a chromosomal region known as the cps locus. The nucleotide sequence in this region has been determined for all serotypes. In contrast, not all CPS structures have been defined. The structure of the serotype 35C polysaccharide was recently reported, but the presence of O-acetyltransferase genes in the serotype 35C cps locus suggested that it could be incomplete, as the reported structure contains no O-acetylation. In addition, the genetic distinction of serotype 35C from the closely related serotype 42 was unclear, as their reported cps loci are nearly identical. To clarify these discrepancies, we obtained serotype 35C and 42 clinical and reference isolates and studied their serological and genetic properties, as well as the structures of CPSs purified from reference isolates. We demonstrated that the O-acetyltransferase WciG was functional in serotype 35C but nonfunctional in serotype 42 due to a deletion in wciG Serotype 35C was O-acetylated at the 5- and 6-positions of 3-?-galactofuranose, as well as the 2-position of 6-?-galactofuranose. However, serotype 42 has only O-acetylation at 3-?-galactofuranose, an observation consistent with its loss of WciG functionality, which is associated with O-acetylation at the 2-position and subsequent reaction with typing antiserum 35a. These findings provide a comprehensive view of the genetic, biochemical structural, and serological bases of serotypes 35C and 42.