Project description:Competition is a major determinant of plant community structure consisting of both species-specific and general interactions, either of which may influence competitive competency and plant abundance and size. In certain cases, competitive competency could arise from altered gene expression and plant function when an individual is confronted with new competitors. We explored competition at the molecular level by hybridizing transcripts from Centaurea maculosa (spotted knapweed), one of North America's most invasive exotic plant species, to an Arabidopsis microarray chip. Centaurea was grown in competition with Festuca idahoensis (Idaho fescue), a native grass species that generally has weak competitive effects against Centaurea; Gaillardia aristata (Indian blanketflower), a native herbaceous species that tends to be a much stronger competitor against Centaurea; or alone (control). The expression of some genes was found to be relatively uninfluenced by the type of plant neighbor, whereas other patterns of gene expression appeared to be more neighbor specific. To our knowledge, these results are the first to identify genes in an invasive plant that are induced or repressed by plant neighbors and provide a new avenue of insight into the molecular aspects of plant competitive ability. Keywords: treated vs.untreated
Project description:Competition is a major determinant of plant community structure consisting of both species-specific and general interactions, either of which may influence competitive competency and plant abundance and size. In certain cases, competitive competency could arise from altered gene expression and plant function when an individual is confronted with new competitors. We explored competition at the molecular level by hybridizing transcripts from Centaurea maculosa (spotted knapweed), one of North America's most invasive exotic plant species, to an Arabidopsis microarray chip. Centaurea was grown in competition with Festuca idahoensis (Idaho fescue), a native grass species that generally has weak competitive effects against Centaurea; Gaillardia aristata (Indian blanketflower), a native herbaceous species that tends to be a much stronger competitor against Centaurea; or alone (control). The expression of some genes was found to be relatively uninfluenced by the type of plant neighbor, whereas other patterns of gene expression appeared to be more neighbor specific. To our knowledge, these results are the first to identify genes in an invasive plant that are induced or repressed by plant neighbors and provide a new avenue of insight into the molecular aspects of plant competitive ability. Keywords: treated vs.untreated First file, Chip 508 (9-9-05) is preliminary hyb data to see if this cross species hybridization is possible using the OAR27K chip Experiment 1 (chip 508): The Arabidopsis OAR27K gene chip was hybridized with labeled cDNA probes produced from samples of Centaurea RNA at the W.M. Keck Foundation Biotechnology Resource Laboratory, Yale University. The OAR27K gene chip consists of 70-mer oligos representing approximately 27,000 genes from Arabidopsis. Leaf and root samples were reverse-transcribed into cDNA and labeled with different fluorescent dyes (Cy3 and Cy5) using the Genisphere Array 900 kit (Cat No.W500180) (Genisphere, Hatfield PA). Labeled cDNAs were hybridized to the OAR27Kchip using the Advalytix ArrayBooster DNA Microarray Incubator (Advalytix, Boston MA). Approximately 10ug of each sample was used for hybridization.Experiment 2: Hybridization and labeling followed the procedure above, except the experimental control was Centaurea plants grown alone, and the test situations were Centaurea grown with Gaillardia or Festuca neighbors. In each experiment, RNA from Centaurea grown alone was used as the control (labeled with Cy3) and RNA from Centaurea grown with a neighbor was used as the test (labeled with Cy5). Two replicates were performed for each test situation.
Project description:Brucella spp. is an intracellular pathogen in vivo. The intracellular B. melitensis transcriptome was determined by initially enriched and then amplified B. melitensis RNA from total RNA of B. melitensis-infected HeLa cells. Analysis of microarray results identified 161 and 115 genes differentially expressed at 4 and 12 h p.i., respectively. Most of the genes (78%) differentially expressed were down-regulated at the earliest time point, but up-regulated (75%) at 12 h p.i. The analysis of the results indicates that Brucella undergo an adaptation period during the first 4 h p.i. that is overcome by 12 h p.i., permitting Brucella to replicate intracellularly. Specific genes and biological processes identified in this study will further help elucidate how Brucella act during the early infectious process to their eventual benefit and to the detriment of the naM-CM-/ve host. Keywords: Time course study of intracellular B. melitensis gene expression Gene expression of the intracellular Brucella melitensis was determined at 4 and 12 h p.i. We generated the following samples: A) B. melitensis total RNA enriched and amplified from total RNA of B. melitensis-infected HeLa cells at 4 h p.i.; B) Total RNA isolated from B. melitensis-infected HeLa cells at 4 h p.i.; C) B. melitensis total RNA enriched and amplified from total RNA of B. melitensis-infected HeLa cells at 12 h p.i.; D) Total RNA isolated from B. melitensis-infected HeLa cells at 12 h p.i. B. melitensis total RNA was initially enriched and then amplified from total RNA of B. melitensis-infected HeLa cells at 4 and 12 h p.i. in quadruplicate, indirectly labeled and co-hybridized against B. melitensis gDNA to a custom 3.2K B. melitensis oligo-array (n = 8). As there was a possibility that some HeLa transcripts cross-hybridize with probes on B. melitensis microarrays, the original total RNA from B. melitensis-infected HeLa cells were also co-hybridized against B. melitensis gDNA to B. melitensis oligo-arrays (n = 8), and any oligospots with signals were considered non-specific and eliminated from all analysis to avoid false positive gene detection. The intracellular B. melitensis gene expression was compared to the gene expression of the inoculum (n = 2). Every Brucella melitensis open reading frame was printed in triplicate on each microarray, thereby providing three technical replicates for each biological replicates. Each replicate was normalized against labeled Brucella melitensis genomic DNA.