Microarray experiment on M. incognita infected and uninfected material in A.thaliana Col-0 plants and A. thaliana ERF6 mutated plants
ABSTRACT: In this experiment we investigate the transcription profile of M. inocognita infection at the start of the infection and 7 days later in A. thalinana Col-0 plants. This was compared to M. incognita infected material of the plant line erf6-1, a T-DNA insertion mutant in ERF6. The data showed the role of ERF6 during early M. incognita infection in Arabidaaaaaaaaaaopsis besides more insight in the transcription regulated by M. incognita in Col-0 the wildtype plant. Samples are taken from whole root systems of 14 (0 days after infection) or 21 (7 days after infection) day old plants.
Project description:affy_meloidogyne_rice2 - affy_meloidogyne_rice2 - Plant-parasitic nematodes cause heavy economic losses to global agriculture. The root knot nematode, Meloidogyne incognita, is an obligate parasite that causes significant damage to a broad range of host plants. M. incognita infection to dicotyledous plants is extensively studied but it is also important to study their interaction with monocotyledous plants, in particular with cereals. In our growing conditions, as of day 6, histological studies revealed a profound rice tissue reorganisation around nematodes, notably characterized by the plant feeding site formation. We are investigating the molecular plant response to M. incognita by carrying out a global analysis of gene expression during gall formation in rice, using giant cell-enriched root tissues at this early stage (6dpi) of gall development-Oryza sativa (var. Nipponbare) seedlings were grown on 6 cm3 SAP substrate completed with diluted Hoaglands solution (Reversat et al., 1999). Culture units were placed in a growth chamber illuminated with fluorescent tubes 9/24 h and maintained at 23°C for 6 days before being inoculated with a 300 J2-stage juveniles M. incognita. One day after inoculation (dai), the rice seedlings were immersed in de-ionised water to remove all J2s that had not penetrated the roots and allowing synchronization of the infection. Each seedling was transferred to a hydroponic mini chamber (Reversat et al., 2004). Sampling was performed at 6 dai and each of them contained galls from 45 infected plants, they were then hand-dissected, frozen in liquid-nitrogen and stored at -80°C. As reference samples, uninfected meristematic root fragments were dissected from seedlings grown under the same conditions. Each sample was replicated 3 times. Keywords: normal vs disease comparison 6 arrays - rice
Project description:Transcript profile of apices of 20 days-old Arabidopsis plants over expressing miR396b. We used ATH1 Affymetrics microarrays to obtain the transcription profile of plants overexpressing miR396b. We compared the transcriptome of 35S:miR396b plants vs wild-type plants (Col-0 accession), using ATH1 Affymetrics microarrays. For these experiments we collected vegetative apices together with the leaf primordia from plants of 20 days growth in short-day. For each genotype (Col-0, 35S:miR396b) we collected two samples.
Project description:111,556,278 high-quality reads were obtained by a deep-sequencing approach that show an exact match to the genome of Meloidogyne incognita from the library of J2 juveniles of M. incognita (Mi). Based on these Solexa reads, 89 M. incognita microRNA genes were identified, which grouped into 67 non-redundant miRNAs with mature sequences. All of these candidate miRNAs were confirmed by qRT-PCR, and 26 of them could be detected in the library of the galls of cucumber root infected by M. incognita (GC). MiR-100 was found in a cluster with let-7, which is similar with B. malayi, a human parasitic nematode. Based on the results of deep sequencing, the expression of miR-100 was much more abundant than that of let-7, which indicated that they may not be co-expressed. The ortholog of let-7, a key regulator that controls the nematode from L4 to adult in C. elegans, could be frequently sequenced in the GC library, the later stages of development of M. incognita, while it had a relative low expression level in J2, which indicated that let-7 may have a similar role in the development regulation in plant parasitic nematodes. Frequently sequenced microRNAs, including miR-71, miR-100 and miR-124, should play an important role in the growth and proliferation of M. incognita. Identifying microRNAs of M. incognita based on deep-sequencing of the small RNAs of an Mi library, the J2 juveniles, and comparing their expression levels with those in a GC library, the gall of cucumber root infected with M. incognita.
Project description:As part of the Cage consortium, leaves 1 and 2 of Col-0 plants were harvested at differents stages of development as defined by Boyes Scale (1.04, 1.12, and 5.10). The Col-0 was used as a control for the coi1-1 plants as both sets were sown on MS media for 7 days and then transferred to soil (Jiffy pots). These samples were hybridized against the oligo reference labeled with Cy3. These samples can be compared to the Col-WT samples that were harvested by the CAGE members to provide a variation of a normal genomic expression pattern.
Project description:By sequencing small RNAs from uninfected Arabidopsis roots and from galls seven and 14 days post infection with Meloidogyne incognita, we sequenced by SOLiD technology the RNA fraction below 50nt. We identified 24 miRNAs differentially expressed in gall as putative regulators of gall development. Overall design: Uninfected Arabidopsis roots and galls seven and 14 days post infection with Meloidogyne incognita (3 replicates)
Project description:As part of the Cage consortium, all leaves (the shoot) of coi1 plants were harvested at the 1.10 stage of development as defined by Boyes scale. The Col-0 was used as a control for the coi1-1 plants as both sets were sown on MS media for 7 days and then transferred to soil (Jiffy pots). These samples were hybridized against the oligo reference labeled with Cy3. These samples can be compared to the Col-WT samples that were harvested by the CAGE members to provide a variation of a normal genomic expression pattern.
Project description:Reactive oxygen species (ROS) have been characterized as both important signaling molecules and universal stressors that mediate many developmental and physiological responses. So far, details of the transcriptional mechanism of ROS-responsive genes are still largely unknown. In the study reported herein, we identified eight potential ROS-responsive cis-acting elements (ROSEs) from the promoters of genes upregulated by ROS. We also found that the APETALA2 (AP2/EREBP)-type transcription factor ERF6 could bind specifically to the ROSE7/GCC box. Co-expression of ERF6 enhanced luciferase activity driven by ROSE7. ERF6 interacted physically with mitogen-activated protein kinase 6 (MPK6), and also served as a substrate of MPK6. MPK6-mediated ERF6 phosphorylation at both Ser 266 and Ser 269 affected the dynamic alternation of ERF6 protein, which resulted in changes in ROS-responsive gene transcription. These data might provide new insight into the mechanisms that regulate ROS-responsive gene transcription via a complex of MPK6, ERF6, and the ROSE7/GCC box. To identifiy the ERF6 regualted genes under ROS and HL treatment. Three-week old Col-0 and erf6-1 mutant plants before and after exposed to 2000 umol m-2s-1 illumination for 2 h were used for RNA extracted hybridization on Affymetrix microarrays.
Project description:Transcriptional profiling of Arabidopsis rossette leaves comparing WT Col-0 with a transgenic lines overexpressing AhNF-YC gene from Amaranthus hypochondriacus in three different conditions. Three-condition experiment WT vs AhNF-YC OE plant leaves. The analyzed conditions were: normal growth conditions, water stress for 5 days and 24 hrs of recovery after normal watering was reestablished to 8-day water stressed plants.
Project description:We have used a strain of Tobacco etch potyvirus (TEV) experimentally adapted to Arabidopsis thaliana ecotype Ler-0 to infect a set of seven A. thaliana plant ecotypes(Col-0, Ei-2, Wt-1, ler-0, Oy-0, St-0). Each ecotype was inoculated with the same amount of the virus. Using commercial microarrays containing probes Arabidopsis thaliana ssp. Col-0 plant transcripts, we explored the effect of viral infection in the plant transcriptome This microarray can be useful to study gene activity of Arabidopsis thaliana associated with response to virus infection. For ecotypes ST-0, Wt-1, Ler-0, Di-2 there are five biological replicates and for ecotype Oy-0 there are three. As control we used four technical replicates of each ecotype, but five for Ler-0. Rows and columns are numbered as scanned by a GenePix Scanner (barcode on the bottom, DNA on the front surface).
Project description:Wild-type (Col-0) and mutant plants (adg1-1, tpt-2, adg1-1/tpt-2) were grown for 28 days under low light conditions and were then transferred to HL. Leaf samples for transcriptional profiling were taken at time 0 (before transfer to high light) and 4 hours and 48 hours (2 days) after transfer to high light.