Project description:Species from the genus Colletotrichum are the causal agents of anthracnose which contribute to significant losses to the production of commercially grown crops. The genomes of Colletotrichum orbiculare, which infects cucurbits and Nicotiana benthamiana, as well as Colletotrichum gloeosporioides, which infects a wide range of fruits and vegetables, were sequenced. A custom microarray was designed for Colletotrichum orbiculare and used to assess gene expression during infection of Nicotiana benthamiana. Gene expression of Colletotrichum orbiculare growing on its host Nicotiana benthamiana was assessed at 24 hours post inoculation, 3 days post inoculation and 7 days post inoculation. Mycelia growing in vitro and ungerminated conidia were used as controls. Three replicates were performed for each time point.
Project description:Time course of C. orbiculare infection of the host plant, N. benthamiana, and comparison to in vitro grown tissue (conidia and in vitro grown hyphae) to identify genes that contribute to the infection process by transcript expression profile.
Project description:Colletotrichum orbiculare Whi2, yeast stress response Whi2 homolog, is involved in switch from biotrophic to necrotrophic stage. To elucidate downstream genes regulated by Co Whi2, we have conducted DNA microarray. About 3100 genes were up or down regulated in the Co whi2Δ mutant compared with the wild-type. In particularly, 44 genes among up-regulated 58 genes in the Co whi2Δ mutant are ribosomal protein related gene. Eukaryote is widely conserved TOR (Target Of Rapamycin) which is known to regulator of ribosomal gene expression. To elucidate whether up-regulated ribosomal genes in the Co whi2Δ mutant are regulated by TOR activity, we have conducted DNA microarray in the Co whi2Δ mutant treated with rapamycin inhibiting TOR activity. The enormous ribosomal gene expression in the Co whi2Δ mutant treated with the rapamycin is lower than that without rapamycin treatment.
Project description:Colletotrichum orbiculare Whi2, yeast stress response Whi2 homolog, is involved in switch from biotrophic to necrotrophic stage. To elucidate downstream genes regulated by Co Whi2, we have conducted DNA microarray. About 3100 genes were up or down regulated in the Co whi2Î mutant compared with the wild-type. In particularly, 44 genes among up-regulated 58 genes in the Co whi2Î mutant are ribosomal protein related gene. Eukaryote is widely conserved TOR (Target Of Rapamycin) which is known to regulator of ribosomal gene expression. To elucidate whether up-regulated ribosomal genes in the Co whi2Î mutant are regulated by TOR activity, we have conducted DNA microarray in the Co whi2Î mutant treated with rapamycin inhibiting TOR activity. The enormous ribosomal gene expression in the Co whi2Î mutant treated with the rapamycin is lower than that without rapamycin treatment. In gene expression of the Co whi2Î mutant, the wild-type and the Co whi2Î mutant infecting on cucumber cotyledons were assessed at 4 hours post-inoculation. In gene expression of the Co whi2Î mutant with rapamycin treatment, Co whi2Î mutant treated with 100nM rapamycin and Co whi2Î mutant without rapamycin treatment infecting on cucumber cotyledons were assesed at 4 hours post-inoculation. Four replication were performed for each experiments.
Project description:Nicotiana benthamiana plants were grown to the 8th true leaf development stage with 16 h light at 150 mol s-1 m-2 with 25/17oC day/night temperatures. Colletotrichum orbiculare strain ATCC20767 and Pseudomonas syringae pv. tabaci strain BPIC4 was grown in pure cultures to obtain syringae pv. tabaci strain BPIC4 was grown in pure cultures to obtain conidia and cells, respectively. Leaves of N. benthamiana plants were inoculated by spraying a suspension of 2 X 106 spores/ml in sterile H2O for C. orbiculare as described by Shen et al. (2001) or by infiltrating with a needle-less syringe 2 X 105 CFU/ml in sterile 10 mM MgCl2 for P. syringae pv. tabaci according to Rommens et al. (1995). For a control, healthy leaves were sprayed with sterile H2O or infiltrated with sterile 10 mM MgCl2 and then incubated under the same conditions for the same period of time as the pathogen-inoculated plants. Total RNA was extracted at 12 hours and 1, 2, 3 and 4 days after inoculation. The first and second mature leaf was excised and immediately frozen at -70 C. Keywords: Direct comparison