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: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:Epidemics of coffee leaf rust (CLR) lead to great yield losses and huge depreciation of coffee marketing values, if no control measures are applied. Societal expectations of a more sustainable coffee production are increasingly imposing the replacement of pesticide treatments by alternative solutions. A good protection strategy is to take advantage of the plant immune system by eliciting its constitutive defenses. Based on such concept, plant resistance inducers (PRIs) have been developed. The Greenforce CuCa formulation made by UFLA (Brazil) is, in addition to acibenzolar-S-methyl (ASM), showing promising results in the control of CLR (Hemileia vastatrix) in Coffea arabica. In order to improve our understanding of the molecular mechanisms of the PRIs, proteomic (2DE-MALDI/TOF/TOF-MS/MS), physiological (leaf gas-exchange) and biochemical (enzymatic) analyses of coffee leaves treated with Greenforce CuCa and ASM and inoculation with H. vastatrix were performed. Proteomic data showed metabolic adjustments mainly related with photosynthesis, protein metabolism and stress responses but, the proteins modulated by the two PRIs were different. Greenforce CuCa, on its own, increased photosynthesis and stomatal conductance, while ASM caused a decrease in these parameters. Upon H. vastratix infection, the Greenforce CuCa showed a higher protective effect on the leaf physiology than ASM. The enzymatic analyses indicated that Greenforce CuCa reinforces the redox homeostasis of the leaf, while ASM seems to increase the involvement of secondary metabolism. So, the PRIs prepare the plant to resist CLR but, inducing different defense mechanisms upon pathogen infection. The data also evidenced the existence of a link between the primary metabolism and defense responses. Furthermore, Greenforce CuCa emerged as a significant agent for CLR management. The identification of components of the plant primary metabolism, essential for plant growth and development that, simultaneously, participate in the plant defense responses can open new perspectives for plant breeding programs.
Project description:Aphid attack induces defense responses in plants activating several signaling cascades that led to the production of toxic, repellent or antinutritive compounds and the consequent reorganization of the plant primary metabolism. Pepper (Capsicum annuum L.) leaf proteomic response against Myzus persicae (Sulzer) has been investigated and analyzed by LC-MS/MS coupled with bioinformatics tools. Infestation with an initially low density (20 aphids/plant) of aphids restricted to a single leaf taking advantage of clip cages resulted in 6 differentially expressed proteins relative to control leaves (3 proteins at 2 days post-infestation and 3 proteins at 4 days post-infestation). Conversely, when plants were infested with a high density of infestation (200 aphids/plant) 140 proteins resulted differentially expressed relative to control leaves (97 proteins at 2 days post-infestation, 112 proteins at 4 days post-infestation and 105 proteins at 7 days post-infestation). The majority of proteins altered by aphid attack were involved in photosynthesis and photorespiration, oxidative stress, translation, protein folding and degradation and amino acid metabolism. Other proteins identified were involved in lipid, carbohydrate and hormone metabolism, transcription, transport, energy production and cell organization. However proteins directly involved in defense were scarce and were mostly downregulated in response to aphids. The unexpectedly very low number of regulated proteins found in the experiment with a low aphid density suggests an active mitigation of plant defensive response by aphids or alternatively an aphid strategy to remain undetected by the plant. Under a high density of aphids, pepper leaf proteome however changed significantly revealing nearly all routes of plant primary metabolism being altered. Photosynthesis was so far the process with the highest number of proteins being regulated by the presence of aphids. In general, at short times of infestation (2 days) most of the altered proteins were upregulated. However, at longer times of infestation (7 days) the protein downregulation prevailed. Proteins involved in plant defense and in hormone signaling were scarce and mostly downregulated.
Project description:Infestation with white-backed planthopper (WBPH) to rice caused induced resistance to rice pathogens but brown planthopper (BPH) infestation induce weaker resistance to rice pathogens. We compared changes in gene expression in rice plants infested with WBPH and BPH to gain some insight into the WBPH-induced resistance to rice pathogens. An analysis, using microarrays, of gene expression in rice plants infested with these planthoppers revealed that WBPH infestation caused high induction of many defense-related genes including pathogenesis-related (PR) genes than BPH infestation. Furthermore, hydroperoxide lyase 2 (OsHPL2) which is an enzyme to produce C6 volatiles was induced by WBPH infestation, but not by BPH infestation. Experiment Overall Design: Agilent rice oligo microarray was used to investigate the gene expression profiling in rice plants infested with WBPH or BPH. Total RNA was extracted from pooled leaf blades infested with WBPH or BPH for 24 h and from mock-treated pooled leaf blades. Total RNA (200 ng) was labeled with Cy-3 or Cy-5 using an Agilent low RNA input linear amplification kit. Fluorescently labeled targets were hybridized to Agilent rice oligo microarrays. Hybridization and wash processes were performed according to the manufacturerâ??s instructions, and hybridized microarrays were scanned using an Agilent DNA microarray scanner. Agilent Feature Extraction software was employed for the image analysis and data extraction processes. Fold changes in expression level in each treatment were compared with those of the respective mock-treated controls. In each treatment, the experiment was performed independently three times.
Project description:Transcriptome analysis in response to infestation of whitefly in peppr leaf and root Microarry study using total RNA from whitefly infestation, BTH, whitefly + BTH, and control in both leaf and root of pepper
Project description:The green rice leafhopper (GRH), Nephotettix cincticeps Uhler, is a major insect pest of cultivated rice in temperate Asia. GRH2-near-isogenic line (NIL) TGRH11, GRH4-NIL (TGRH16) and GRH2/GRH4-pyramided line (PYL) TGRH29 were developed by introducing the GRH2 and GRH4 from indica rice (DV85). We identified GRH-inducible genes in respective rice lines. Furthermore, we compared the gene expression levels between NILs/PYL and control plants (T65). The gene expression changes in respective rice lines were detected by comparison between GRH-infested and pre-infested plants. Seedlings at the second-leaf stage were infested with 10 to 15 first- or second-instar nymphs in test tubes and shoots were collected at 30 h after GRH infestation. Detached-leaf blades at heading stage were infested with 10 to 15 first- or second-instar nymphs in test tubes and samples were collected at 30 h after GRH infestation. For each treatment, two biological replicates were performed.