ABSTRACT: Comparative transcript profiling of gene expression between seedless Ponkan mandarin (Citrus reticulata Blanco) and its seedy wild type during floral organ development
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.
Project description:Seed developmental arrest is one of the early phenotypes of seed abortion. However, the molecular mechanism underlying seed developmental arrest of citrus is still unclear. In this study, laser capture microdissection (LCM) was used to accurately divide the seeds of seedless Ponkan ‘Huagan No.4’ (Citrus reticulata) (HG) and seeded Ponkan ‘Egan No.1’ (Citrus reticulata) (EG) into nucellus and integument/seed coat tissues. The captured tissues were used for subsequent RNA-seq. Moreover, single-molecule real-time (SMRT) sequencing was used to generate full-length transcripts of EG, which were used as reference transcripts for RNA-seq. These data can be utilized to analyse the causes of citrus seedlessness formation and the molecular regulatory network in the process of seed abortion.
Project description:Ovule developmental arrest is one of the early phenotypes of seed abortion. However, the molecular mechanism underlying ovule developmental arrest of citrus is still unclear. In this study, laser capture microdissection (LCM) was used to accurately divide the ovules of seedless Ponkan ‘Huagan NO.4’ (Citrus reticulata) (MT) and seeded Ponkan ‘Egan NO.1’ (Citrus reticulata) (WT) into nucellus and integument tissues. The captured tissues were used for subsequent RNA-seq. Moreover, single-molecule real-time (SMRT) sequencing was used to generate full-length transcripts of WT, which were used as reference transcripts for RNA-seq. These data can be utilized to analyse the causes of citrus seedlessness formation and the molecular regulatory network in the process of ovule abortion.
Project description:Seedlessness is an important agronomic trait for citrus, and male sterility (MS) is one main cause of seedless citrus fruit. However, the molecular mechanism of citrus seedlessness remained not well explored. An integrative strategy combining SSH (suppression subtractive hybridization) library with cDNA microarray was employed to study the underlying mechanism of seedlessness of a Ponkan mandarin seedless mutant (Citrus reticulate Blanco). Screening with custom microarray, a total of 279 differentially expressed clones were identified, and 133 unigenes (43 contigs and 90 singletons) were obtained after sequencing. Gene Ontology (GO) distribution showed that the majority of differential genes involved in metabolic process, responded to stimulus and functioned as DNA/RNA binding, catalytic activity and oxidoreductase activity. Expression dynamics of some candidate clones revealed a gene encoding male sterility protein 2 was highly up-regulated, while several transcription factors (TFs) such as AP2/EREBP, MYB, WRKY, NAC and C2C2-GATA zinc-finger domain TFs were specifically down-regulated in the seedless mutant compared with the wild type. Our research highlighted some candidate pathways that participated in the citrus male gametophyte development and could be beneficial for seedless citrus breeding in the future.
Project description: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. âPonkanâ 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:Seedlessness is an important agronomic trait for citrus, and male sterility (MS) is one main cause of seedless citrus fruit. However, the molecular mechanism of citrus seedlessness remained not well explored. An integrative strategy combining SSH (suppression subtractive hybridization) library with cDNA microarray was employed to study the underlying mechanism of seedlessness of a Ponkan mandarin seedless mutant (Citrus reticulate Blanco). Screening with custom microarray, a total of 279 differentially expressed clones were identified, and 133 unigenes (43 contigs and 90 singletons) were obtained after sequencing. Gene Ontology (GO) distribution showed that the majority of differential genes involved in metabolic process, responded to stimulus and functioned as DNA/RNA binding, catalytic activity and oxidoreductase activity. Expression dynamics of some candidate clones revealed a gene encoding male sterility protein 2 was highly up-regulated, while several transcription factors (TFs) such as AP2/EREBP, MYB, WRKY, NAC and C2C2-GATA zinc-finger domain TFs were specifically down-regulated in the seedless mutant compared with the wild type. Our research highlighted some candidate pathways that participated in the citrus male gametophyte development and could be beneficial for seedless citrus breeding in the future. Flower buds from the seedless citrus mutant and its wild type were collected at four developmental stages: quaring stage (SF, about 20 DBF), medium bud stage (MF, about 10 DBF), flowers at full bloom stage (BF) and young ovaries of 2-3 days after flowering (OV). Total RNA extracted from the mutant and the wild type was hybridized to the array. The hybridization was performed in duplicate by dye swap.
Project description:High-throughput small RNA and degradome sequencing were performed to identify a large number of miRNAs and their targets. Differential expression of several miRNAs reveals that miRNA regulatory network was closely linked to gametophyte development and male sterility in Ponkan mandarin. The expression profiles of miR156 and miR167 as well as their targets demonstrated their contribution in male sterility in the Ponkan seedless type. This study provides valuable information for better understanding of miRNAs network in plant anther development and lays a foundation to future research on male sterility in citrus and even other species.
Project description:High-throughput small RNA and degradome sequencing were performed to identify a large number of miRNAs and their targets. Differential expression of several miRNAs reveals that miRNA regulatory network was closely linked to gametophyte development and male sterility in Ponkan mandarin. The expression profiles of miR156 and miR167 as well as their targets demonstrated their contribution in male sterility in the Ponkan seedless type. This study provides valuable information for better understanding of miRNAs network in plant anther development and lays a foundation to future research on male sterility in citrus and even other species. small RNA and degradome sequencing of seedy M-bM-^@M-^XEgan No.1M-bM-^@M-^Y and its male sterile and seedless mutant M-bM-^@M-^XQianyang seedlessM-bM-^@M-^Y
