ABSTRACT: The postharvest senescence processes of citrus fruits were analyzed transcriptomic. The present study was aimed to: further uncover the rind-flesh communication of hesperidium; characterize the differential storage behaviors of different citrus varieties; reveal the important changes during storing process; and demonstrate the specific non-climacteric characteristics of citrus fruits. We chose four major table fruit varieties of citrus: satsuma mandarin (Citrus unshiu Marc) (M), ponkan (Citrus reticulata Blanco) (K), newhall navel orange (Citrus sinensis L. Osbeck) (O) and shatian pummelo (Citrus grandis Osbeck) (P). They were sampled every 10 days during 50 DAH (days after harvest), almost covering the commercial storage period of loose-skin citrus.
Project description:Low temperature storage of citrus fruits are susceptible to chilling disorder symptoms that impact fruit quality. Understanding the molecular frame underlying the cold storage process will provide a basic guidance for practical control. We used Affymetrix Citrus GeneChip to examine the transcriptional changes in cold-stored Citrus reticulata Blanco cv. ‘Ponkan’ pulp tissue for three successive months. Ponkan postharvest fruits were under cold storage for three successive months. The pulp tissue was used for RNA extraction and hybridization on Affymetrix Citrus Genome microarrays.
Project description:Low temperature storage of citrus fruits are susceptible to chilling disorder symptoms that impact fruit quality. Understanding the molecular frame underlying the cold storage process will provide a basic guidance for practical control. We used Affymetrix Citrus GeneChip to examine the transcriptional changes in cold-stored <i>Citrus reticulata</i> Blanco cv. ‘Ponkan’ pulp tissue for three successive months. Overall design: Ponkan postharvest fruits were under cold storage for three successive months. The pulp tissue was used for RNA extraction and hybridization on Affymetrix Citrus Genome microarrays.
Project description:An antifungal aroma substance, 2-phenylethanol (PEA), was isolated from antagonistic yeast strain Kloeckera apiculata extract. Microarry were used to analyse its role citrus. We used microarrays to detail the global programme of gene expression underlying Citrus were treated with 1.0x108 cells/ml K. apiculata (KA), PEA (0.15%), the extract (1000xdilute) and control (CK) for 24 h, An antifungal aroma substance, 2-phenylethanol, was isolated from antagonistic yeast strain Kloeckera apiculata. To analyse its role in Citrus response,Citrus were treated with K. apiculata , 2-phenylethano (0.15%), the extract (1000xdilute) and control (CK) for 24 h, respectively. The fresh epicarp of citrus was separated by knife and directly frozen in liquid nitrogen for RNA extraction and hybridization on Affymetrix microarrays.
Project description:Kumquat (Fortunella classifolia Swingle) was thought as a close relative to citrus according to fruit morphology taxonomic, but kumquat fruit as well as its flowering characteristic are distinct from other citrus species, the trees usually blooms on the medium of June, obviously later than other citrus species, moreover many kumquat accessions could be blossom more than one times during one growth season, as this reason, the kumquat fruits could be consecutively ripen over several months, which made the study of globe genes expression profile for different development stage fruit easy. Kumquat is non-climacteric fruit, however the kumquat fruit ripening process, especially genes expression change in young and ripe kumquat fruits are less known, so studying on the global genes expression profiles of kumquat fruits in young and ripe stage are especially helpful to identify ripening-related genes and unravel the mechanism of ripening process.
Project description:Somatic embryogenesis in nucellar tissues is widely recognized to induce polyembryony in major citrus varieties such as sweet oranges, satsuma mandarins and lemons. This capability for apomixis is attractive in agricultural production systems using hybrid seeds, and many studies have been performed to elucidate the molecular mechanisms of various types of apomixis. To identify the gene responsible for somatic embryogenesis in citrus, a custom oligo-DNA microarray including predicted genes in the citrus polyembryonic locus was used to compare the expression profiles in reproductive tissues between monoembryonic and polyembryonic varieties. The full length of CitRKD1, which was identified as a candidate gene responsible for citrus somatic embryogenesis, was isolated from satsuma mandarin and its molecular function was investigated using transgenic ‘Hamlin’ sweet orange by antisense-overexpression. The candidate gene CitRKD1, predominantly transcribed in reproductive tissues of polyembryonic varieties, is a member of the plant RWP-RK domain proteins. CitRKD1 of satsuma mandarin comprised two alleles (CitRKD1-mg1 and CitRKD1-mg2) at the polyembryonic locus controlling embryony type (mono/polyembryony) that were structurally divided into two types with or without a miniature inverted-repeat transposable element (MITE)-like insertion in the upstream region. CitRKD1-mg2 with the MITE insertion was the predominant transcript in flowers and young fruits where somatic embryogenesis of nucellar cells occurred. Loss of CitRKD1 function by antisense-overexpression abolished somatic embryogenesis in transgenic sweet orange and the transgenic T1 plants were confirmed to derive from zygotic embryos produced by self-pollination by DNA diagnosis. Genotyping PCR analysis of 95 citrus traditional and breeding varieties revealed that the CitRKD1 allele with the MITE insertion (polyembryonic allele) was dominant and major citrus varieties with the polyembryonic allele produced polyembryonic seeds. Overall design: To identify the gene responsible for somatic embryogenesis in citrus, a custom oligo-DNA microarray including predicted genes in the citrus polyembryonic locus was used to compare the expression profiles in reproductive tissues between ‘Kiyomi’ and ‘Harumi’ as materials for monoembryonic and polyembryonic varieties, respectively. Eight independent experiment performed at 15, 30, 45 and 60 DAF (Day After Flowering) using whole young fruits in each variety.
Project description:Bud mutations arise often in citrus. The selection of mutants is one of the most important breeding methods in citrus. However, the molecular bases of bud mutation have rarely been studied. To identify the potential important or novel genes involved in bud mutation, different transcriptomic techniques combing suppression subtractive hybridization (SSH) and microarray were performed between a lycopene accumulated mutant, ‘Hong Anliu’ sweet orange (Citrus sinensis L. Osbeck), and its wild-type during fruit maturation. Microarray analysis revealed that differentially expressed clones are extensively coordinated with the initiation of lycopene accumulation. After sequencing of the differentially expressed clones, a total of 267 non-redundant transcripts were obtained, 182 (68.2%) of which share homology (E-value ≤ 1×10-10) with known gene products or known protein domains. A list of candidate genes which involved in cellular metabolic process, primary metabolic process, localization, macromolecular metabolic process was obtained. Out of these genes, 12 share homology with previously described signal transduction or transcription factors, suggesting complex regulatory control. These results demonstrate profound effect on gene expression of bud mutation in citrus fruits and provide new insights into the molecular basis of bud mutation. Keywords: bud mutation, candidate genes, Citrus, cNDA microarray, suppression subtractive hybridization (SSH) Fruits from the mutant and its wild type were collected at five time points from August to December. Total RNA extracted from the mutant was hybridized to the array together with RNA from the wild type. Each hybridization was performed in duplicate by dye swap.
Project description:Storage at low temperatures is one of the most used methods to prolong the life of postharvest peaches (Prunus persica (L) Batch.). However, fruit quality is adversely affected by the development of woolliness, a physiological disorder that is apparent when the fruit is ripened after prolonged periods of cold storage and is mainly manifested as loss of juice in the peaches. The aim of this study was to obtain a more detailed cohort of genes that underlie the wolliness in a segregating population with contrasting phenotypes of mealiness after being exposed to cold storage at 4 °C. For this, a transcriptomics approach was applied to fruits from a progeny of individuals accounted for 6% more juicy and woolly 6% over a 2 years. Our results suggest that not only genes related to the maintenance of cell wall architecture may contribute to the development of mealy phenotype. Based on its possible physiological process and differential pattern of expression transcriptomic profiles show that genes related to maintenance (modification I) and membrane fluidity account for the differences between fruits that exhibit contrasting phenotypes of mealiness. These genes may contribute to tolerance to cold during storage. We analyzed a total of 9 woolly fruits (from 3 different trees, 3 fruits from each tree) and 12 juicy fruits (from 4 different trees, 3 fruits from each tree). An RNA pool from 9 woolly fruits was used as reference and was compared to an RNA pool of 3 juicy fruits from each individual tree. Two technical replicates were done for each comparison, thus making in total 8 hybridizations.
Project description:Mature green fruits of the tomato (Solanum lycopersicum L.) cultivar MicroTom were investigated (fruit developmental category II). After harvest, fruits were immediately utilized or stored in darkness at 25 C until further processing. After 0, 1, 2 or 4 days of storage stem scar tissues of tomato fruits were excised with a scalpel to a depth of approximately 2 mm, immediately frozen in liquid nitrogen and stored at -80 C until use.
Project description:Transcript profiling analysis of fruits from a segregating PopDG population, which were grouped/selected according to extreme cold susceptibility and subjected to different times of cold storage
Project description:The tomato MADS-box FRUITFULL (FUL) homologs, FUL1 and FUL2, interact with the main ripening regulator RIPENING INHIBITOR (RIN). To clarify their role in fruit ripening, we generated FUL1/FUL2-suppressed transgenic lines by RNAi. We found that five transgenic lines bearing fruits that did not ripen normally: lycopene accumulation and increase of ethylene production were severely inhibited. We then performed next generation RNA sequencing (RNA-Seq) analysis of the fruits of a FUL1/FUL2-suppressed line (TF18) with those of the wild type (Ailsa Craig cultivar; AC) and rin mutant. The comparison of RNA-Seq data among them indicated that FUL1/FUL2-suppression significantly affected the expression of a larger portion of ripening-induced and -repressed genes than the rin mutation did. Moreover, the effect of FUL1/FUL2-suppression was observed not only in the fruits harvested at the wild type ripening age [45 days after pollination (DAP)] but also in those at the pre-ripening age (35 DAP). This suggests that the FUL homologs play an essential role in the regulation of fruit development and ripening, the role which covers a wider range of biological processes than RIN does. Differentially expressed genes (DEGs) between the wild type and TF18 fruits included known ripening-related genes such as ACS2 and ACS4 involved in ethylene production and PSY1 in carotenoid biosynthesis, consistent with the phenotype of TF18 fruits described above. The DEGs also included many direct RIN target genes, which supports the hypothesis that the FUL homologs regulate fruit ripening in a form of MADS-box complex with RIN. mRNA profiles of wild type (Ailsa Craig cultivar), rin mutant and FUL1/FUL2-suppressed tomato fruits harvested at 35DAP and 45 DAP were generated by next generation sequencing, in triplicate, using Illumina Hiseq2000.