Project description:Background: Understanding the genetic elements that contribute to key aspects of coffee biology will impact future agronomical improvements for this economically important tree. The past years, EST collections were generated in Coffee, opening the possibility to create new tools for functional genomics. Results: The project PUCE CAFE, set up by the scientific consortium NESTLE/IRD/CIRAD has developed of long oligonucleotide coffee array using public coffee EST sequences mainly obtained from different stages during fruit development and leaves in Coffea canephora (Robusta). We have performed a validation experiment in order to check the array usability and the reproducibility of hybridizations. Conclusion: We have generated the first 15K Coffee array during this three years project PUCE CAFE, granted by The French National Research Agency (ANR, Programme Génoplante) . This new tool was dedicated to large scale transcriptomic analysis during grain development of Coffea canephora grown in different countries . Furthermore, other analysis have been also initiated by the different partners like analysis of polyploidy or drought resistance. In any case, at the end of the project, the generated arrays will be available to the international scientific community.
Project description:The present study was conducted to evaluate the effects of the intake of three types of coffee (caffeinated, decaffeinated, and green unroasted coffee) on the livers of C57BL/6J mice fed a high-fat diet, and to extensively elucidate the physiological responses to coffee intake by analysing the findings obtained from a comprehensive transcriptomic analysis using DNA microarrays. The present study was conducted to evaluate the effects of the intake of three types of coffee (caffeinated, decaffeinated, and green unroasted coffee) on the livers of C57BL/6J mice fed a high-fat diet, and to extensively elucidate the physiological responses to coffee intake by analysing the findings obtained from a comprehensive transcriptomic analysis using DNA microarrays.
Project description:Background: Understanding the genetic elements that contribute to key aspects of coffee biology will impact future agronomical improvements for this economically important tree. The past years, EST collections were generated in Coffee, opening the possibility to create new tools for functional genomics. Results: The project PUCE CAFE, set up by the scientific consortium NESTLE/IRD/CIRAD has developed of long oligonucleotide coffee array using public coffee EST sequences mainly obtained from different stages during fruit development and leaves in Coffea canephora (Robusta). We have performed a validation experiment in order to check the array usability and the reproducibility of hybridizations. Conclusion: We have generated the first 15K Coffee array during this three years project PUCE CAFE, granted by The French National Research Agency (ANR, Programme Génoplante) . This new tool was dedicated to large scale transcriptomic analysis during grain development of Coffea canephora grown in different countries . Furthermore, other analysis have been also initiated by the different partners like analysis of polyploidy or drought resistance. In any case, at the end of the project, the generated arrays will be available to the international scientific community. three biological replicates were made for each tissue analyzed (i.e. leaves, flowers and mature beans). The following comparisons were made: Bean-Flower, Leaf-Flower and Leaf-Bean. In all, we performed microarray analyses on 18 slides [3 (replicates) x 2 (dyes) x 3 (organs)]
Project description:Coffee leaf miner is an important plague in coffee crops. Using subtracted cDNA libraries and nylon filter arrays, we analyzed the expression profile of 1536 expressed sequence tags (ESTs) of coffee plants from an hybrid progeny (C. arabica x C. racemosa), containg resistant (R) and susceptible plants (S) to the infestation of coffee leaf miner. Leaf discs were collected from non-infested plants (R control - RC; S control - SC), infested plants after moth oviposition (R oviposition - Ro; S oviposition - So) and infested after larvar eclosion (R eclosion - Re; S eclosion - Se). Isolation and characterization of Coffea genes induced during coffee leaf miner (Leucoptera coffeella) infestation. Plant Science 169(2):351-360 Keywords: ordered
Project description:The present study was conducted to evaluate the effects of the intake of three types of coffee (caffeinated, decaffeinated, and green unroasted coffee) on the livers of C57BL/6J mice fed a high-fat diet, and to extensively elucidate the physiological responses to coffee intake by analysing the findings obtained from a comprehensive transcriptomic analysis using DNA microarrays. The present study was conducted to evaluate the effects of the intake of three types of coffee (caffeinated, decaffeinated, and green unroasted coffee) on the livers of C57BL/6J mice fed a high-fat diet, and to extensively elucidate the physiological responses to coffee intake by analysing the findings obtained from a comprehensive transcriptomic analysis using DNA microarrays. Briefly, 7-week-old male C57BL/6J mice purchased from Charles River Laboratories Japan (Yokohama) were divided into the following five groups. The normal diet group (ND group) was fed D12450B (10 kcal% fat, Research Diets, New Brunswick, NJ, USA). The high-fat diet group (HF group) was fed D12492 (60 kcal% fat, Research Diets, New Brunswick, NJ, USA). The caffeinated coffee group (HFCC group) was fed a high-fat diet containing 2% caffeinated freeze-dried coffee. The decaffeinated coffee group (HFDC group) was fed a high-fat diet containing 2% decaffeinated freeze-dried coffee. The green unroasted coffee group (HFGC group) was fed a high-fat diet containing 2% unroasted caffeinated freeze-dried coffee. The mice had ad libitum access to their diets and drinking water. After 9 weeks, mice were sacrificed and the livers were subjected to the Affymrtix DNA microarray experiment.
Project description:The MADS-domain transcription factor APETALA1 (AP1) is a key regulator of Arabidopsis flower development. To understand the molecular mechanisms underlying AP1 function, we identified its target genes during floral initiation using a combination of gene expression profiling and genome-wide binding studies. Many of its targets encode transcriptional regulators, including known floral repressors. The latter genes are down-regulated by AP1, suggesting that it initiates floral development by abrogating the inhibitory effects of these genes. While AP1 acts predominantly as a transcriptional repressor during the earliest stages of flower development, regulatory genes known to be required for floral organ formation were found to be activated by AP1 at more advanced stages, indicating a dynamic mode of action. Our results further imply that AP1 orchestrates floral initiation by integrating growth, patterning and hormonal pathways.
Project description:Characterization of the activities of the transcription factor that AG encodes throughout flower development using perturbation assays and ChIP-Seq in combination with a floral induction system (FIS) that allows a stage-specific analysis of flower development. Examination of genomic regions bound by fully functional AG-GFP protein at approx floral stage 4-5 as compared to a negative control sample.
Project description:Coffee leaf miner is an important plague in coffee crops. Using subtracted cDNA libraries and nylon filter arrays, we analyzed the expression profile of 1536 expressed sequence tags (ESTs) of coffee plants from an hybrid progeny (C. arabica x C. racemosa), containg resistant (R) and susceptible plants (S) to the infestation of coffee leaf miner. Leaf discs were collected from non-infested plants (R control - RC; S control - SC), infested plants after moth oviposition (R oviposition - Ro; S oviposition - So) and infested after larvar eclosion (R eclosion - Re; S eclosion - Se). Isolation and characterization of Coffea genes induced during coffee leaf miner (Leucoptera coffeella) infestation. Plant Science 169(2):351-360
Project description:Characterization of the activities of the transcription factors that AP3 and PI encode throughout flower development using perturbation and ChIPSeq assays in combination with a floral induction system (FIS) that allows a stage-specific analysis of flower development. Examination of genomic regions bound by fully functional AP3-GFP and PI-GFP proteins at approx floral stage 4-5 as compared to a negative control sample
Project description:The MADS-domain transcription factor APETALA1 (AP1) is a key regulator of Arabidopsis flower development. To understand the molecular mechanisms underlying AP1 function, we identified its target genes during floral initiation using a combination of gene expression profiling and genome-wide binding studies. Many of its targets encode transcriptional regulators, including known floral repressors. The latter genes are down-regulated by AP1, suggesting that it initiates floral development by abrogating the inhibitory effects of these genes. While AP1 acts predominantly as a transcriptional repressor during the earliest stages of flower development, regulatory genes known to be required for floral organ formation were found to be activated by AP1 at more advanced stages, indicating a dynamic mode of action. Our results further imply that AP1 orchestrates floral initiation by integrating growth, patterning and hormonal pathways. We used the AP1-GR system to conduct chromatin immunoprecipitation experiments with AP1-specific antibodies followed by deep-sequencing (ChIP-Seq) in order to determine AP1 binding sites on a genome-wide scale. Samples were generated from tissue in which the AP1-GR protein was induced for 2h using a single treatment of 1 uM DEX to the shoot apex. As control, we performed ChIP experiments using the same antibody on uninduced tissue. Experiments were done in two biological replicates.