Project description:Transcriptome analysis in tobacco mutant plants using tomato Genechip Genome array Tobacco (Nicotiana tabacum cv. Petit Havanna) psaA and psbA deletion mutants were constructed through a targeted deletion of 767 and 1152 nucleotides of coding regions, respectively with two gene cassettes: psbAproR:uidA:psbterR and rrnR:aadA:rbcLterR coding for GUS reporter and spectinomycin selectable marker genes, respectively. Standard established procedures were followed for chloroplast transformation to generate the psaA and psbA deletion mutants based on the homologous recombination. Gene expression profiles in psaA and psbA tobacco mutant plants were analyzed using tomato Genechip Genome array to study the global changes in the expression of genome.
Project description:Transcriptome analysis in tobacco mutant plants using tomato Genechip Genome array Tobacco (Nicotiana tabacum cv. Petit Havanna) psaA and psbA deletion mutants were constructed through a targeted deletion of 767 and 1152 nucleotides of coding regions, respectively with two gene cassettes: psbAproR:uidA:psbterR and rrnR:aadA:rbcLterR coding for GUS reporter and spectinomycin selectable marker genes, respectively. Standard established procedures were followed for chloroplast transformation to generate the psaA and psbA deletion mutants based on the homologous recombination. Gene expression profiles in psaA and psbA tobacco mutant plants were analyzed using tomato Genechip Genome array to study the global changes in the expression of genome. Total RNA was isolated from psaA and psbA tobacco mutant plants along with the wild type plants. Biotin labeled cRNA was hybridized on tomato GeneChip Genome Array following the Affymetrix protocols. Two independent biological replicates were maintained.
Project description:Transgenic tobacco (Nicotiana tabacum) expressing Caenorhabditis elegans cell death genes, Ced4 and Ced3, show evidence suggesting such expressions protect the plants from infestation by the plant parasitic nematode Meloidogyne incognita. Although positive results have been correlated with the gene expressions (data in preparation for publication; a draft of the publication can be provided upon request), the mechanism by which the nematode protection is manifested is not clearly understood. One possibility is that the C. elegans cell death proteins produced by the transgenic plants are being ingested and incorporated into the nematode’s own cell death pathway, leading to their demise. Alternatively, it is also possible that expression of the C. elegans cell death genes promotes the endogenous resistance genes of the plant, leading to nematode resistance. We want to test the later hypothesis by conducting a reference design microarray experiment to establish the expression profile of Ced3, and Ced4 homozygous plants and Ced3xCed4 double heterozygous plants in comparison with wild-type tobacco plants. If the hypothesis is correct, we expect to detect increased expression of pathogenicity-related genes in the transgenic plants. Furthermore, characterization of the expression profiles in these transgenic plants will provide us directionality for our future research on the elucidation of this resistance mechanism. Keywords: Reference design
Project description:Gene expression was measured in leaves from dark treated tobacco plants to investigate the changes associated with dark induced senescence.
Project description:Comparative transcriptome analysis was performed to study gene expression profiles in resistant (Yanyan 97, YY97, 25) and susceptible (Huanghuadajinyuan, HD, 36) tobacco in responding to Ralstonia solanacearum infection. Illumina sequencing yielded a total of 67,619,833,668 bases data, and about 223.99 M and 223.82 M raw reads for Hd and Yy97 plants, respectively. About 209.73 M and 209.18 M clean reads of Hd and Yy97 were mapped to reference genome via Hisat2, respectively. The ratio of mapped clean reads for eight libraries ranged from 93.92% to 96.67% (average: 95.9%). By comparing gene expression levels in Rs infected and control tobacco stems, we identified 15374 DEGs in Hd plants after Rs infection, which included 7220 up-regulated and 8154 down-regulated DEGs. We identified 2120 DEGs in Yy97 plants after Rs infection, which included 1794 up-regulated and 326 down-regulated DEGs.
Project description:Transgenic tobacco (Nicotiana tabacum) expressing Caenorhabditis elegans cell death genes, Ced4 and Ced3, show evidence suggesting such expressions protect the plants from infestation by the plant parasitic nematode Meloidogyne incognita. Although positive results have been correlated with the gene expressions (data in preparation for publication; a draft of the publication can be provided upon request), the mechanism by which the nematode protection is manifested is not clearly understood. One possibility is that the C. elegans cell death proteins produced by the transgenic plants are being ingested and incorporated into the nematode’s own cell death pathway, leading to their demise. Alternatively, it is also possible that expression of the C. elegans cell death genes promotes the endogenous resistance genes of the plant, leading to nematode resistance. We want to test the later hypothesis by conducting a reference design microarray experiment to establish the expression profile of Ced3, and Ced4 homozygous plants and Ced3xCed4 double heterozygous plants in comparison with wild-type tobacco plants. If the hypothesis is correct, we expect to detect increased expression of pathogenicity-related genes in the transgenic plants. Furthermore, characterization of the expression profiles in these transgenic plants will provide us directionality for our future research on the elucidation of this resistance mechanism. Keywords: Reference design 27 hybs total
Project description:Genome-wide selection and testing of superior reference genes for quantitative gene expression normalization in tobacco (Nicotiana tabacum)