A Putative Early Response of Antifungal Bacillus lentimorbus WJ5 Against the Plant Pathogenic Fungus, Colletotrichum gloeosporioides, Analyzed by a DNA Microarray
The global RNA transcription profiles of Bacillus lentimorbus WJ5 under an in vitro co-culture with Colletotrichum gloeosporioides were analyzed in order to study the antagonistic bacteria-fungi interactions. Using a filter membrane system, B. lentimorhus WJ5 was exposed to the spores of C. gloeospo...
The global RNA transcription profiles of Bacillus lentimorbus WJ5 under an in vitro co-culture with Colletotrichum gloeosporioides were analyzed in order to study the antagonistic bacteria-fungi interactions. Using a filter membrane system, B. lentimorhus WJ5 was exposed to the spores of C. gloeosporioides at the late exponential stage. The transcription profiles of the B. lentimorhus WJ5, both with and without a challenge from C. gloeosporioides, were analyzed using custom DNA chips containing 2,000 genome fragments. A total of 337 genes were expressed, with 87 and 47 up- and down-regulated, respectively. Of these, 12 genes, which were involved in central carbon metabolisms, and 7 from minor catabolism were relatively highly up-regulated (> 10 fold) and down-regulated (< 0.2 fold), respectively. Nine genes, which were thought to be related to the antifungal activity, were also up-regulated, but their levels were not so high (2.0 - 9.7 folds). From the results, during the early stage of the co-culture of B. lentimorbus WJ5 and C. gloeosporioides, nutrient competition seemed to occur; therefore, the genes from central carbon metabolisms could be up-regulated, while those from minor catabolism could be down-regulated.
The global RNA transcription profiles of Bacillus lentimorbus WJ5 under an in vitro co-culture with Colletotrichum gloeosporioides were analyzed in order to study the antagonistic bacteria-fungi interactions. Using a filter membrane system, B. lentimorhus WJ5 was exposed to the spores of C. gloeosporioides at the late exponential stage. The transcription profiles of the B. lentimorhus WJ5, both with and without a challenge from C. gloeosporioides, were analyzed using custom DNA chips containing 2,000 genome fragments. A total of 337 genes were expressed, with 87 and 47 up- and down-regulated, respectively. Of these, 12 genes, which were involved in central carbon metabolisms, and 7 from minor catabolism were relatively highly up-regulated (> 10 fold) and down-regulated (< 0.2 fold), respectively. Nine genes, which were thought to be related to the antifungal activity, were also up-regulated, but their levels were not so high (2.0 - 9.7 folds). From the results, during the early stage of the co-culture of B. lentimorbus WJ5 and C. gloeosporioides, nutrient competition seemed to occur; therefore, the genes from central carbon metabolisms could be up-regulated, while those from minor catabolism could be down-regulated.
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제안 방법
In this work, custom DNA microarrays of the antago nistic bacterium, B. lentimorbus WJ5, were constructed and the RNA transcription profiles from the in vitro co culture of B. lentimorbus WJ5 against C. gloeosporioides observed, in order to study the antagonist-fungus interaction.
Poor quality spots (sum of median < 300) were filtered from the raw data before analysis. The microarray experiments were repeated twice, and the average of the two gene expres- sion values for each gene used for the analysis. For each hybridized spot, the gene expression value (ratio of the median in the GenePix Pro 3.
대상 데이터
The RNA of the B. lentimorbus WJ5 was labeled with cyanine 3-dUTP (Cy3-dUTP, NEN Life Science Products, USA) and 나sed as the control probe. The RNA of the B.
이론/모형
This study was carried out under the Nuclear R&D Pro gram of the Ministry of Science and Technology (MOST), Republic of Korea.
성능/효과
The microarray experiments were repeated twice, and the average of the two gene expres- sion values for each gene used for the analysis. For each hybridized spot, the gene expression value (ratio of the median in the GenePix Pro 3.0), 나le ratio of the Cy5 flu orescence intensity minus the Cy5 background intensity to that of the Cy3 fluorescence intensity minus the Cy5 background intensity, was calculated. This represented the gene expression fold change for each gene.
From the results of our experiments, nutrient competi tion seemed to occur at an early stage of the co-culture of B. lentimorbus WJ5 and C. gloeosporio ides, and in advance of antibiosis, so that the genes from a central car bon metabolism could be up-regulated and those from a minor metabolism could be down-regulated.
후속연구
, 1999). However, further study on the func tions of the 6 unknown 응enes (K233, K1215, K1688, K1547, K1757 and K1221) appears to be required to establish any relationship to nutrient competition.
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