Transcription profiling by array of Andigena SP5G RNAi leaves, tubers and tuber sprouts
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ABSTRACT: To analyze the gene regulatory network controlled by SP5G in potato, we hybridized an Agilent 60-mer microarray with probes corresponding to the RNA extracted out of leaf, tuber and tuber sprout tissues from WT and StSP5Gi lines.
Project description:To analyze the gene regulatory network controlled by SP6A under warm temperatures, we hybridized an Agilent 60-mer microarray with probes corresponding to the RNA extracted out of leaves from WT and StSP6Aox potato plants grown at 22ºC and 28ºC.
Project description:Transcriptome Analysis of the potato (genotype RH89-039-16). To aid annotation and address a series of biological questions, we generated RNA-Seq data from 16 RH libraries representing all major tissue types, developmental stages and responses to abiotic and biotic stresses.
Project description:Potato (Solanum tuberosum L) is a natural host of Potato spindle tuber viroid (PSTVd) which can cause characteristic symptoms on developing plants including stunting phenotype and distortion of leaves and tubers. PSTVd is the type species of the family Pospiviroidae, it can replicate in the nucleus and the viroid RNA moves systemically in infected plants. Its KF440-2 strain can cause severe symptoms in potato. It is not well understood how the viroid can affect host genes for successful invasion and which genes show altered expression levels upon infection. In this study, we used a high-scale method to identify differentially expressed genes in potato. We have identified defence, stress and sugar metabolism related genes having altered expression levels upon infection. Additionally, hormone pathways connected genes showed up- or down-regulation. Our primary focus is on the identification of genes which can affect tuber formation as the viroid infection can strongly influence tuber development, especially tuber shape is affected. DWARF1/DIMINUTO, Gibberellin 7-oxidase and BEL5 protein were identified and validated which showed differential expression in viroid infected tissues suggesting that gibberellin and brassinosteroid pathways have a possible role in tuber development upon PSTVd infection.
Project description:In Cichorium intybus, macroscopic and histological modifications were identified during in vitro cell reactivation and morphogenesis events. The relationships between these modifications and transcript profiles of genes were investigated. Two chicory genotypes derived from the Hungarian landrace Koospol, were tested: K59 is a responsive genotype (embryogenic), as it undergoes cell de- and re-differentiation that leads to somatic embryogenesis (SE). C15 is an unresponsive genotype (nonembryogenic), unable to express this morphogenetic pattern. Previously studies showed that addition of beta-D-glucosyl Yariv reagent (BGlcY), a synthetic phenyl-glycoside that specifically binds arabinogalactan-proteins (AGPs), to the culture medium blocked somatic embryogenesis in chicory root explants in a concentration-dependent manner with complete inhibition of induction occurring at 250 µM of BGlcY (Chapmann et al. 2000a). A putative AGP (DT212818) encoding gene was previously found significantly over-expressed in the K59 as compared to the C15 and was suggested to be involved in chicory re-differentiation. To better understand the cell reactivation events, a treatment with BGlcY was applied in order to precipitate AGPs in muro and to block the in vitro cell re-differentiation processes. The two different genotypes together with BGlcY represented an original tool to discriminate cell reactivation events from cell fate determination. Microscopy, biochemical and transcriptome analyses showed that in vitro SE in chicory was blocked during BGlcY-treatment but cell reactivation still occurred. Consequently, AGP (DT212818) seems to be only important in cell fate determination leading to SE commitment. Gene expression profiles showed in addition to the AGP, other proteins could play a role in SE determination: 26S proteasome AAA ATPase subunit 6 (RPT6), remorin (REM), metallothionein 1 (MT1), 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase). Cell wall events during BGlcY-treatment were observed and discussed. Gene expression of K59 explants induced (d4) was compared with gene expression of K59 explants without induction (d0). Gene expression of K59 explants induced in the presence of BGlcY (Y+; d4) was next compared with gene expression of K59 explants induced in the absence of BGlcY (Y-; d4). Similar comparisons were made for C15 leaf explants. To link up both genotypes, direct comparison was done between gene expression of K59 explants without induction (d0) and C15 explants without induction (d0). Three biological replicates were used for each analysed condition. Dye labelling for each paired sample was reversed in two subsequent individual hybridizations giving rise to a total of six hybridizations per condition except for the K59_D0_vs_C15_D0 condition.
Project description:The present research investigates a ‘medicinal’ plant Jerusalem artichoke (abbreviated as JA) (Helianthus tuberosus L.) tuber proteome with an aim to unravel its proteome using a high-throughput proteomics technique. Although JA has been historically know to the Native Americans, it was brought back and spread to Europe by the colonists and in the late 19th century early 20th century it began to regain importance including its use for health and as a folk remedy for diabetes. In Japan (referred to as ‘kiku-imo’) its cultivation became popular mostly for health-related benefits such as reducing the blood sugar level. The group of (Genboku Takahashi et al.) has been working on the cultivation and utilization of kiku-imo tuber as a traditional/alternative medicine in daily life, and thus the research progressed to deeply look into the protein components through proteomics as very less is known about the proteome of the tubers, especially in relation to its importance as a functional food in treating diseases health conditions like diabetes. Using three commercially processed JA tuber products we used total protein extraction on the powdered samples in conjunction with label-free quantitate proteomic approach (mass spectrometry) to identify for the first time a comprehensive protein list for the JA tuber. A total of 2967 high‐confidence proteins were identified and categorized into different protein classes through bioinformatics. We have discussed these proteins especially in relation to their association with health and disease regulatory metabolism.
Project description:72 plants have been grown in 2 phytochambers for 30 days under control temperature (21°C/19°C) and short day conditions (8 hours light/16 hours darkness). After 30 days the plants were switched to long day conditions (16 hours light/8 hours darkness) and the temperature of one phytochamber was increased to 29°C/27° (heat chamber). Moreover,a heatplate was located in the control chamber and a coldplate was located in the heat chamber. 18 plants have been grown on the heatplate with a constant temperature of 29°C. 18 plants have been grown on the coldplate with a constant temperature of 22°C. After 9 days of the heat period leaf samples (end of night) were taken for micorarray analysis. After 10 days of heat, tuber samples were taken for microarray analysis.
Project description:Leaf explants of the superembryogenic Medicago truncatula line 2HA were treated with auxin (1-naphthaleneacetic acid) for one week to induce the formation of roots (Imin et al J Exp Bot 58:439-451). Gene expression in the leaves and the NAA treated tissue cultures was compared to identify transcripts expressed during the commitment to root formation in tissue culture. We have used the Affymetrix Medicago Genome Array GeneChip to compared gene expression in Medicago truncatula leaves and leaf explants that have been cultured for one week on NAA, to identify genes expressed during the commitment to root formation in tissue culture. Experiment Overall Design: Medicago truncatula 2HA leaves and leaf explants treated with NAA for one week were collected; total RNA was extracted and used for hybridization to Affymetrix arrays
Project description:The consequences on tuber transcriptome of a short heat period during tuber development was investigated in this study with special regard to the development of secondary tuber growth. Plants were grown for 47 days in the greenhouse under ambient conditions (21°C/ 19°C, 16h light, 8h dark) before application of mild heat stress temperatures (29°C/27°C) to one group of plants for 7 days and a stress release period on control temperature for 2 more weeks until harvest. Leaves were sampled before the heat period, at the end of the heat period and at harvest, two weeks after stress release. Tuber samples were taken at harvest. Tubers grown at normal temperatures and exhibiting a normal growth phenotype were used as control. Tubers subjected to the heat treatment and exhibiting a second-growth phenotype (chain tubers) were grouped into primary (attached to stolon from plant) and secondary tubers (attached to stolon from primary tuber).
Project description:In order to identify cellular proteins interacting with the coat protein (CP) of Potato virus Y (PVY), we used coimmunoprecipitation of GFP-CP in PVY-infected Nicotiana benthamiana plants. To be able to pull down non-abundant interactors and to stabilize transient interactions, we used the crosslinker dithiobis(succinimidyl propionate) (DSP). With this we were able to identify a total of 147 potential CP interactors.