Regulation of cuticle formation during fruit development and ripening in 'Newhall' navel orange (Citrus sinensis Osbeck) revealed by transcriptomic and metabolomic profiling
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ABSTRACT: To investigate global genome expression changes during fruit developmemt, We then performed genome-wide expression analyses on the fruit peel at 120 and 150 DAF with RNA-seq approach.A total of 865 genes were identified as differentially expressed, among which 494 genes were up-regulated and 371 genes were down-regulated in the fruits harvested at 150 DAF compared with in those harvested at 120 DAF.In general, data obtained from RNA-seq showed that the genes involved in photosynthesis and tetrapyrrole synthesis were significantly repressed at 150 DAF; in contrast, the genes involved in secondary metabolism including the biosynthesis of wax, terpenoids and phenylpropanoid were significantly induced at 150 DAF. Total four sample pools (two independent biological replicates of fruits peel from 120 DAF and 150 DAF)
Project description:To investigate global genome expression changes during fruit developmemt, We then performed genome-wide expression analyses on the fruit peel at 120 and 150 DAF with RNA-seq approach.A total of 865 genes were identified as differentially expressed, among which 494 genes were up-regulated and 371 genes were down-regulated in the fruits harvested at 150 DAF compared with in those harvested at 120 DAF.In general, data obtained from RNA-seq showed that the genes involved in photosynthesis and tetrapyrrole synthesis were significantly repressed at 150 DAF; in contrast, the genes involved in secondary metabolism including the biosynthesis of wax, terpenoids and phenylpropanoid were significantly induced at 150 DAF.
Project description:Peel color is a key factor that affects the fruit’s aesthetic and economic values. In Red Sugar pineapple, the peels’ red color reduces during maturation. Limited knowledge is available on the regulation of pineapple peel discoloration, which makes it important to study the molecular mechanisms associated with this important trait. Here, we report that a decrease in anthocyanin biosynthesis is predominantly associated with the pineapple peel color change during maturation. Particularly the exclusive accumulation of cyanidin in 60 days after flowering (DAF) as compared to 120 DAF gives the fruit peel its distinct reddish color. Our findings suggest that the changes in the expression of key structural genes (early and late biosynthetic genes) of the anthocyanin (cyanidin) biosynthesis pathway are responsible for peel discoloration. Based on a gene co-expression analysis and a transient expression, we identified two transcription factors i.e., AcHOX21 and AcMYB12, and showed that their downregulation leads to the reduced anthocyanin accumulation with fruit maturation.
Project description:The quality of the pepper fruit is significantly influenced by the properties of its surface such as color, glossiness and texture. The fruit surface is composed of a peel containing several layers including the cuticle, epidermis and the hypodermis. The peel acts as a protective barrier against biotic and abiotic stresses and is the most critical tissue affecting water loss during post harvest storage. The peel is composed of an outer epidermis with thick waxy (lipid) cuticle and few cell layers of thick-walled hypodermal cells. Despite its agronomic importance and due to the fact that the majority of studies in fruits have been conducted using flesh and peel tissues as a whole, the biochemical and genetic bases of variation in peel properties are largely unknown. In this proposal we aim to determine peel-specific gene expression in pepper by micro array hybridizations of peel and flesh RNA extracted at different developmental stages of the fruit. The cultivar Celica (Capsicum annuum) that has a large blocky fruit will be used for studying gene expression in the peel and flesh. Plants were grown in the greenhouse during the spring of 2006. Fruits were harvested at three developmental stages: young- 10 days after anthesis, mature green- 30 days after anthesis and ripe red- 45 days after anthesis. These stages were chosen because each represents a distinct phase in fruit development. At each stage, a biological replicate consists of bulked tissue from 3 fruits from each of 3 plants (a total of 9 fruits). We have a total of 4 biological replicates. For each fruit, the peel was separated from the flesh by manual dissection using thin forceps and scalpel blade. Peel and flesh samples were immediately frozen in liquid nitrogen and stored at -800C until RNA extraction. Total RNA was extracted using the GenElute Mammalian Total RNA Miniprep kit (Sigma). Keywords: Reference design 12 hybs total
Project description:The quality of the pepper fruit is significantly influenced by the properties of its surface such as color, glossiness and texture. The fruit surface is composed of a peel containing several layers including the cuticle, epidermis and the hypodermis. The peel acts as a protective barrier against biotic and abiotic stresses and is the most critical tissue affecting water loss during post harvest storage. The peel is composed of an outer epidermis with thick waxy (lipid) cuticle and few cell layers of thick-walled hypodermal cells. Despite its agronomic importance and due to the fact that the majority of studies in fruits have been conducted using flesh and peel tissues as a whole, the biochemical and genetic bases of variation in peel properties are largely unknown. In this proposal we aim to determine peel-specific gene expression in pepper by micro array hybridizations of peel and flesh RNA extracted at different developmental stages of the fruit. The cultivar Celica (Capsicum annuum) that has a large blocky fruit will be used for studying gene expression in the peel and flesh. Plants were grown in the greenhouse during the spring of 2006. Fruits were harvested at three developmental stages: young- 10 days after anthesis, mature green- 30 days after anthesis and ripe red- 45 days after anthesis. These stages were chosen because each represents a distinct phase in fruit development. At each stage, a biological replicate consists of bulked tissue from 3 fruits from each of 3 plants (a total of 9 fruits). We have a total of 4 biological replicates. For each fruit, the peel was separated from the flesh by manual dissection using thin forceps and scalpel blade. Peel and flesh samples were immediately frozen in liquid nitrogen and stored at -800C until RNA extraction. Total RNA was extracted using the GenElute Mammalian Total RNA Miniprep kit (Sigma). Keywords: Reference design
Project description:Superficial scald is a major physiological disorder in apple fruit that is induced by cold storage and is mainly expressed as brown necrotic patches on peel tissue. However, a global view of the gene-protein-metabolite interactome underlying scald prevention/sensitivity is currently missing. Herein, we have found for the first time that cold storage in an atmosphere enriched with ozone (O3) induced scald symptoms in ‘Granny Smith’ apple fruits during subsequent ripening at room temperature. In contrast, treatment with the ethylene perception inhibitor 1-methylcyclopropene (1-MCP) reversed this O3-induced scald effect. Amino acids, including branched-chain amino acids, were the most strongly induced metabolites in peel tissue of 1-MCP treated fruits. Proteins involved in oxidative stress and protein trafficking were differentially accumulated prior to and during scald development. Genes involved in photosynthesis, flavonoid biosynthesis and ethylene signaling displayed significant alterations in response to 1-MCP and O3. Analysis of regulatory module networks identified putative transcription factors (TFs) that could be involved in scald. Subsequently, a transcriptional network of the genes-proteins-metabolites and the connected TFs was constructed. This approach enabled identification of several genes co-regulated by TFs, notably encoding glutathione S-transferase (GST) protein(s) with distinct signatures following 1-MCP and O3 treatments. Overall, this study is an important contribution to future functional studies and breeding programs for this fruit, aiding to the development of improved apple cultivars to superficial scald.
Project description:Fruits of the tomato (Solanum lycopersicum L.) cultivar MicroTom were investigated. MicroTom wild type and lecer6 mutant fruits with a deficiency in a fatty acid beta-ketoacyl-CoA synthase (LeCER6) were used. Identification and characterization of this insertional mutant has been reported previously (Vogg et al., 2004; Leide et al., 2007). According to the developmental stage of the wild type and lecer6 mutant fruits, samples were composed of 5 to 15 whole fruits with seeds being removed (fruit developmental category 'fruit set' and I). Otherwise, samples contained exclusively the fruit peel, which was removed with a scalpel to a depth of approximately 1 mm (fruit developmental category II to VII). After sampling point the plant material was immediately frozen in liquid nitrogen and stored at -80 C until use.
Project description:To identify genes associated with citrus peel development and manifestation of peel disorders, we analyzed flavedo, albedo and juice sac tissues from navel orange displaying, and not displaying, the puff disorder. Symptomatic and healthy M-bM-^@M-^\NavelM-bM-^@M-^] orange fruits were harvested from an orchard located in in Pauma Valley, San Diego County, California, USA. Sampling for all analysis (healthy or disordered Navel orange) was performed at the same time, from trees grown under the same agronomic, soil, and environmental conditions. Healthy and disordered fruits were analyzed at the mature stage. All transcriptome analysis was performed on mature fruit. For each type of fruit, three tissues (flavedo, albedo, and juice sacs) from three different trees (biological replicates) were separately analyzed. Four symptomatic fruits comprised one biological replicate each. Two healthy fruits comprised two biological replicates of control samples. A 1 cm-thick equatorial disc and four sections (N, S, E, and W) were cut per fruit. Each section of flavedo, albedo, and juice sac tissue was dissected. gene expression variation underlying quality trait, different genotypes
Project description:Anthocyanins are high value plant antioxidants which are not present in the fruits of cultivated tomato. However, both the dominant gene Anthocyanin fruit (Aft) and the recessive gene atroviolacea (atv), introgressed into domesticated tomato from two different wild Solanum species, stimulate a limited anthocyanin pigmentation. Surprisingly, double mutant Aft/Aft atv/atv tomatoes are characterised by the presence of anthocyanins in the fruit peel, resulting in intensely purple pigmented fruit. We carried out a transcript profiling analysis using GeneChip® Tomato Genome Arrays, in order to identify differentially expressed genes when comparing wild type, Aft/Aft, atv/atv, and Aft/Aft atv/atv fruits. The expression pattern of several genes involved in the anthocyanin pathway was analyzed in detail. Among the fruit peel-associated differentially expressed transcripts, genes involved in phenylpropanoid pathway, cell wall composition, biotic and abiotic stress responses, sugar and hormone metabolism were overrepresented in Aft/Aft atv/atv. Transcriptomic analysis thus revealed that the activation of anthocyanin synthesis in tomato fruit was accompanied by a complex remodulation of gene expression, likely affecting important agronomic and merceological traits.
Project description:Mature green fruits of the tomato (Solanum lycopersicum L.) cultivar John Baer and Pearson were investigated (fruit developmental category II). John Baer wild type, John Baer LA0063 and Pearson wild type, Pearson 2-303 fruits were used. Tomato mutant plants LA0063 and 2-303 were positional sterile (PS mutants). Samples contained exclusively the fruit peel, which was removed with a scalpel to a depth of approximately 1 mm. After sampling point the plant material was immediately frozen in liquid nitrogen and stored at -80 C until use.
Project description:Anthocyanins are high value plant antioxidants which are not present in the fruits of cultivated tomato. However, both the dominant gene Anthocyanin fruit (Aft) and the recessive gene atroviolacea (atv), introgressed into domesticated tomato from two different wild Solanum species, stimulate a limited anthocyanin pigmentation. Surprisingly, double mutant Aft/Aft atv/atv tomatoes are characterised by the presence of anthocyanins in the fruit peel, resulting in intensely purple pigmented fruit. We carried out a transcript profiling analysis using GeneChip® Tomato Genome Arrays, in order to identify differentially expressed genes when comparing wild type, Aft/Aft, atv/atv, and Aft/Aft atv/atv fruits. The expression pattern of several genes involved in the anthocyanin pathway was analyzed in detail. Among the fruit peel-associated differentially expressed transcripts, genes involved in phenylpropanoid pathway, cell wall composition, biotic and abiotic stress responses, sugar and hormone metabolism were overrepresented in Aft/Aft atv/atv. Transcriptomic analysis thus revealed that the activation of anthocyanin synthesis in tomato fruit was accompanied by a complex remodulation of gene expression, likely affecting important agronomic and merceological traits. Wild type (Cv. Ailsa Craig, accession number LA2838A), Aft/Aft (accession number LA1996), atv/atv (accession number LA0797) and double mutant (Aft/Aft atv/atv) were grown during the winter season in a controlled heated greenhouse. Fruits were collected at mature green, turning red and red stages of development. The transcriptional profile in Aft/Aft, atv/atv, and Aft/Aft atv/atv fruits when compared to the wild type was analyzed using the GeneChip® Tomato Genome Array.