Project description:Aim of the project: Genome wide gene expression profiles across the cambial zone are analyzed in 35um resolution from wild type hybrid aspen (Populus tremula x tremuloides) and two independent LMX5::AtIPT7 over expressor transgenic Populus tree lines.
Project description: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. Results Treatment of the aspen roots with 500 µM Al induced a strong inhibition of root growth within 6 h of exposure time. The root growth subsequently recovered, reaching growth rates comparable to that of control plants. Changes in gene expression were determined after 6 h, 2 d, and 10 d of Al exposure. Replicated transcriptome analyses using the Affymetrix poplar genome array revealed a total of 175 significantly up-regulated and 69 down-regulated genes, of which 70% could be annotated based on Arabidopsis genome resources. Between 6 h and 2 d, the number of responsive genes strongly decreased from 202 to 26, and then the number of changes remained low. The responses after 6 h were characterized by genes involved in cell wall modification, ion transport, and oxidative stress. Two genes with prolonged induction were closely related to the Arabidopsis Al tolerance genes ALS3 (for Al sensitive 3) and MATE (for multidrug and toxin efflux protein, mediating citrate efflux). Patterns of expression in different plant organs and in response to Al indicated that the two aspen genes are homologs of the Arabidopsis ALS3 and MATE. Conclusion Exposure of aspen roots to Al results in a rapid inhibition of root growth and a large change in root gene expression. The subsequent root growth recovery and the concomitant reduction in the number of responsive genes presumably reflect the success of the roots in activating Al tolerance mechanisms. The aspen genes ALS3 and MATE may be important components of these mechanisms.
Project description:Background: Solubilized aluminum (Al) is rhizotoxic and can be present in acid soils at levels that inhibit root growth. Acid soils are found throughout the world and often support forests. However, unlike in herbaceous plants, little is known about the mechanisms by which forest trees respond to and tolerate Al. To begin to elucidate these mechanisms, we characterized transcriptomic changes in response to Al in roots of aspen (Populus tremula L.). Results: Aspen roots were treated with Al in solution culture for 6 h, 54 h, and 246 h. Transcriptomic changes were assessed by the Affymetrix GeneChip Poplar Genome Array. The analysis revealed 176 induced and 66 suppressed genes. The majority of these genes were regulated at 6 h, presumably reflecting root growth, which was strongly inhibited at 6 h and partially was recovered at 2 d and 10 d. Enrichment analysis identified sets of functionally related genes whose members were statistically over-represented compared to the genes on the microarray. These sets included genes related to cell wall modification, oxidative stress, cell death, and transport processes. Two of the genes involved in transport were related to the Arabidopsis Al tolerance genes AtALS3 (aluminum sensitive 3), possibly mediating redistribution of Al, and AtMATE (multi-drug and toxin extrusion), facilitating exudation of citrate. Expression patterns in response to Al in different plant tissues indicated that the two aspen genes are homologs of AtALS3 and AtMATE, suggesting that aspen and Arabidopsis share common mechanisms to cope with Al. Conclusion: This is the first survey of transcriptomic changes in response to Al in a forest tree. The results of our study provide a valuable set of data, which will help to further our understanding of the mechanisms and their regulation that enable aspen to grow in environments with toxic levels of Al. Aspen roots were treated with Al in solution culture for 6 h, 54 h, 246 h to cover a broad range of exposure times. Transcriptomic changes were assessed by the Affymetrix GeneChip Poplar Genome Array. For each time point, root tips of three independant plants were analyzed.
Project description:Thermospermine-induced transcriptomic changes were explored in Populus tremula x P. tremuloides. Transgenic hybrid aspen plants expressing 35S::POPACAULIS5 were compared to wild-type hybrid aspen under the influence of PGRs and depleted from PGRs.
Project description:Despite being located at the crossroads of Asia, genetics of the Afghanistan populations have been largely overlooked. It is currently inhabited by five major ethnic populations: Pashtun, Tajik, Hazara, Uzbek and Turkmen. Here we present autosomal from a subset of our samples, mitochondrial and Y- chromosome data from over 500 Afghan samples among these 5 ethnic groups. This Afghan data was supplemented with the same Y-chromosome analyses of samples from Iran, Kyrgyzstan, Mongolia and updated Pakistani samples (HGDP-CEPH). The data presented here was integrated into existing knowledge of pan-Eurasian genetic diversity. The pattern of genetic variation, revealed by structure-like and Principal Component analyses and Analysis of Molecular Variance indicates that the people of Afghanistan are made up of a mosaic of components representing various geographic regions of Eurasian ancestry. The absence of a major Central Asian-specific component indicates that the Hindu Kush, like the gene pool of Central Asian populations in general, is a confluence of gene flows rather than a source of distinctly autochthonous populations that have arisen in situ: a conclusion that is reinforced by the phylogeography of both haploid loci. 24 samples were analysed with the Illumina platform Human660W-Quad v1.0 Genotyping BeadChip and are described herein.
Project description:Despite being located at the crossroads of Asia, genetics of the Afghanistan populations have been largely overlooked. It is currently inhabited by five major ethnic populations: Pashtun, Tajik, Hazara, Uzbek and Turkmen. Here we present autosomal from a subset of our samples, mitochondrial and Y- chromosome data from over 500 Afghan samples among these 5 ethnic groups. This Afghan data was supplemented with the same Y-chromosome analyses of samples from Iran, Kyrgyzstan, Mongolia and updated Pakistani samples (HGDP-CEPH). The data presented here was integrated into existing knowledge of pan-Eurasian genetic diversity. The pattern of genetic variation, revealed by structure-like and Principal Component analyses and Analysis of Molecular Variance indicates that the people of Afghanistan are made up of a mosaic of components representing various geographic regions of Eurasian ancestry. The absence of a major Central Asian-specific component indicates that the Hindu Kush, like the gene pool of Central Asian populations in general, is a confluence of gene flows rather than a source of distinctly autochthonous populations that have arisen in situ: a conclusion that is reinforced by the phylogeography of both haploid loci.
Project description:Aim of the project: Genome wide gene expression analysis for cytokinin (100nM 2ip, 20nM NaPi buffer) fast (1h) response genes from 3 months old Hybrid aspen(Populus tremula x tremuloides)apical part (30 cm from tip; diameter:4-5mm) of stem. Stems of two individuals (trees 15 and 16) was cut into 50-100um thick (free hand)cross sections randomly selected stem discs are set to two pools: cytokinin treatment and control treatment. Samples are collected noon time (11:00-11:30). Cytokinin treatment: stem discs (several hundred) are submerged with 100nM 2ip, 20nM NaPi buffer, with modest shaking for 60 min +/- 2 min (time to collect about 30mg stem discs (several dozens) for RNA sample. Control treatment: identical to cytokinin treatment, only without 100nM 2ip.
Project description:The expression of stress-related genes induced by feeding of chestnut moth larvae (Conistra vaccinii L.) was studied with Vitreoscilla hemoglobin-expressing (VHb) and non-transgenic hybrid aspen lines (Populus tremula x P. tremuloides). Besides the herbivore-injured leaves (L1), cDNA microarray analyses were conducted using uninjured leaves of hybrid aspen lines positioned above (A) and below (B) the herbivory exposed leaves.
Project description:In this study we report on transgenic hybrid aspen (Populus tremula x P. tremuloides) lines that showed potential to increase biomass production both in the greenhouse and after five years of growth in the field. The transgenic lines carried an overexpression construct for Populus tremula x tremuloides vesicle-associated membrane protein (VAMP)-associated protein PttVAP27-17 (Potri.019G116400) that was selected from a gene-mining program for novel regulators of wood formation. A proteomic analysis was performed to characterize the overall effect of the overexpression of PttVAP27-17 on plant metabolic pathways. 20 mg of xylem sample for the selected wild type (WT) and three transgenic lines (lines 1,2 and 3) was collected in the following manner from two-months-old, greenhouse grown trees: The frozen bottom-part of the stem (10-17 cm portion from the base of the stem) was peeled, and the surface of the secondary xylem consisting of living vessels and fibers (into the depth of approximately one mm from the surface) was scraped. The xylem scrapings were ground to fine powder in liquid nitrogen and stored at -80oC. The analyses included 3-5 biological replicates for each of the transgenic lines, and seven replicates for the wild type.