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: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:Transgenic expression of viral proteins in natural host plants is a useful simplified system with the potential to understand the individual effect of each viral component. Transgenic expression of movement (MP) and a variant from coat protein (CPT42W) in tobacco, a TMV natural host, produces severe morphological changes, altered miRNAs accumulation and poor fertility. We used microarrays to characterize the gene expression changes caused by the co-expression of TMV capsid and movement proteins in Nicotiana tabacum comparing two isogenic lines MPxCPT42W and mpxcpT42W* (a line with both transgenes spontaneously silenced and with normal phenotype). Leaf tissues from 6-week old tobacco plants MPxCPT42W and mpxcpT42W* were collected for RNA extraction and hybridization on Affymetrix microarrays. We collected pools of three plants (one biological replicate) and analyzed three independent biological replicates for each transgenic line.
Project description:Gene expression was measured in leaves from dark treated tobacco plants to investigate the changes associated with dark induced senescence.
Project description:Two tobacco transgenic lines over-expressing grapevine polygalacturonase-inhibiting protein (Vvpgip1) vs. WT tobacco under normal growth conditions
Project description:Transgenic expression of viral proteins in natural host plants is a useful simplified system with the potential to understand the individual effect of each viral component. Transgenic expression of movement (MP) and a variant from coat protein (CPT42W) in tobacco, a TMV natural host, produces severe morphological changes, altered miRNAs accumulation and poor fertility. We used microarrays to characterize the gene expression changes caused by the co-expression of TMV capsid and movement proteins in Nicotiana tabacum comparing two isogenic lines MPxCPT42W and mpxcpT42W* (a line with both transgenes spontaneously silenced and with normal phenotype).
Project description:Cell adaptation to high salinity levels implicates the modification of different cellular, physiological and molecular mechanisms. However, studies about the cellular mechanisms that are implicated in salt adaptation are scarce in the literature. Tobacco BY-2 cell cultures are very homogeneous and are characterized by a continuous cell grow and high proliferation rate. These features make these cells a good model system. For this study, we have obtained a stable tobacco cell line adapted to grow at 250 mM of NaCl. To obtain a general view of this process, we have followed a microarray technique. Gene expression was analyzed using the Affymetrix microarray technique. We have used a microarray designed by Edwards et al. (2010) in tobacco (Total probes: 43768).
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.