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PROVIDER: PRJNA1105020 | ENA |
REPOSITORIES: ENA
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Project description:Purpose: de novo sequencing and comparative analysis of the bark transciptomes of Hevea brasiliensis induced without ethephon (C), with ethephon for 8 hours (E8) and 24 hours (E24) to identify the genes and pathways related to the stimulation of rubber production by ethylene. The goals of this study are to reveal the molecular mechanism behind the stimulation of rubber production by ethylene. Methods: Bark RNA was extracted using the TRIzol® Reagent (Invitrogen) and two cDNA libraries, H (healthy rubber trees) and T (TPD-affected trees), were prepared using the mRNA-Seq 8 sample prep Kit (Illumina). The libraries were deep sequenced using Illumina HiSeqTM 2000 (Illumina Inc., San Diego, CA, USA). Raw reads produced from sequencing machines were resorted to de novo assembly and gene annotation. Results: De novo sequencing and assembly of the bark transciptomes of Hevea brasiliensis induced with ethephon for 8 hours (E8) and 24 hours (E24) were performed. 51,965,770, 52,303,714 and 53,177,976 high-quality clean reads from E8, E24 and C (control) samples were assembled into 81,335, 80,048 and 80,800 unigenes respectively, with a total of 84,425 unigenes and an average length of 1,101 bp generated. 10,216 and 9,374 differentially expressed genes (DEGs) in E8 and E24 compared with C were respectively detected. The expression of several enzymes in crucial points of regulation in glycolysis were up-regulated and DEGs were not significantly enriched in isopentenyl diphosphate (IPP) biosynthesis pathway. In addition, up-regulated genes of great regulatory importance in carbon fixation (Calvin cycle) were identified. Conclusions: The rapid acceleration of glycolytic pathway supplying precursors for the biosynthesis of IPP and natural rubber, instead of rubber biosynthesis per se, may be responsible for ethylene stimulation of latex yield in rubber tree. The elevated rate of flux throughout the Calvin cycle may account for some durability of ethylene-induced stimulation. Our finding lays the foundations for molecular diagnostic and genetic engineering for high-yielding improvement of rubber tree. De novo sequencing of the transcriptomes of C (bark without ethephon application), E8 (bark with 1.5%-ethephon treatment for 8 hours) and E24 (bark with 1.5%-ethephon treatment for 24 hours) rubber trees was conducted using Illumina HiSeq 2000.
2016-02-23 | E-GEOD-78145 | biostudies-arrayexpress
Project description:Purpose: de novo sequencing and comparative analysis of the bark transciptomes of Hevea brasiliensis induced without ethephon (C), with ethephon for 8 hours (E8) and 24 hours (E24) to identify the genes and pathways related to the stimulation of rubber production by ethylene. The goals of this study are to reveal the molecular mechanism behind the stimulation of rubber production by ethylene. Methods: Bark RNA was extracted using the TRIzol® Reagent (Invitrogen) and two cDNA libraries, H (healthy rubber trees) and T (TPD-affected trees), were prepared using the mRNA-Seq 8 sample prep Kit (Illumina). The libraries were deep sequenced using Illumina HiSeqTM 2000 (Illumina Inc., San Diego, CA, USA). Raw reads produced from sequencing machines were resorted to de novo assembly and gene annotation. Results: De novo sequencing and assembly of the bark transciptomes of Hevea brasiliensis induced with ethephon for 8 hours (E8) and 24 hours (E24) were performed. 51,965,770, 52,303,714 and 53,177,976 high-quality clean reads from E8, E24 and C (control) samples were assembled into 81,335, 80,048 and 80,800 unigenes respectively, with a total of 84,425 unigenes and an average length of 1,101 bp generated. 10,216 and 9,374 differentially expressed genes (DEGs) in E8 and E24 compared with C were respectively detected. The expression of several enzymes in crucial points of regulation in glycolysis were up-regulated and DEGs were not significantly enriched in isopentenyl diphosphate (IPP) biosynthesis pathway. In addition, up-regulated genes of great regulatory importance in carbon fixation (Calvin cycle) were identified. Conclusions: The rapid acceleration of glycolytic pathway supplying precursors for the biosynthesis of IPP and natural rubber, instead of rubber biosynthesis per se, may be responsible for ethylene stimulation of latex yield in rubber tree. The elevated rate of flux throughout the Calvin cycle may account for some durability of ethylene-induced stimulation. Our finding lays the foundations for molecular diagnostic and genetic engineering for high-yielding improvement of rubber tree.
2016-02-23 | GSE78145 | GEO
Project description:Ethylene is commonly used as a latex stimulant of Hevea brasiliensis by application of ethephon (chloro-2-ethylphosphonic acid); however, the molecular mechanism by which ethylene increases latex production is not clear. To better understand the effects of ethylene stimulation on the laticiferous cells of rubber trees, a latex expressed sequence tag (EST)-based complementary DNA microarray containing 2973 unique genes (probes) was first developed and used to analyze the latex gene expression changes at three different time-points after ethephon treatment: 8, 24 and 48 h. Transcript levels of 163 genes were significantly altered with fold-change values ≥ 2 or ≤ –2 (q-value < 0.05) in ethephon-treated compared with control rubber trees. Of the 163 genes, 92 were up-regulated and 71 down-regulated. The microarray results were further confirmed using real-time quantitative reverse transcript-PCR for 20 selected genes.
2016-06-01 | GSE74060 | GEO
Project description:Ethylene is commonly used as a latex stimulant of Hevea brasiliensis by application of ethephon (chloro-2-ethylphosphonic acid); however, the molecular mechanism by which ethylene increases latex production is not clear. To better understand the effects of ethylene stimulation on the laticiferous cells of rubber trees, a latex expressed sequence tag (EST)-based complementary DNA microarray containing 2973 unique genes (probes) was first developed and used to analyze the latex gene expression changes at three different time-points after ethephon treatment: 8, 24 and 48 h. Transcript levels of 163 genes were significantly altered with fold-change values ⥠2 or ⤠â2 (q-value < 0.05) in ethephon-treated compared with control rubber trees. Of the 163 genes, 92 were up-regulated and 71 down-regulated. The microarray results were further confirmed using real-time quantitative reverse transcript-PCR for 20 selected genes. Analysis used the 8, 24 or 48 h control latex RNA samples comparison to the ET stimulated 8, 24 or 48 h latex RNA samples. Each sample included three independent biological replicates, and each replicate comprised the latex collected from six trees.
2016-06-01 | E-GEOD-74060 | biostudies-arrayexpress
Project description:Nutural rubber (NR) production, latex is harvested by periodical tapping of the trunk bark. Ethylene enhances and prolongs latex flow and latex regeneration. Ethephon, which is an ethylene-releasing compound, applied to the trunk before tapping usually results in a 1.5- to 2-fold increase in latex yield. We investigated gene expression in response to ethephon treatment using Pará rubber tree seedlings as a model system. After ethephon treatment, 3,270 genes showed significant differences in expression compared with the mock treatment. Genes associated with carotenoids, flavonoids, and abscisic acid biosynthesis were significantly upregulated by ethephon treatment, which might contribute to an increase in latex flow. Genes associated with secondary cell wall formation were downregulated, which might be because of the reduced sugar supply. Given that sucrose is an important molecule for NR production, a trade-off may arise between NR production and cell wall formation for plant growth and for wound healing at the tapping panel.
2021-09-22 | GSE174832 | GEO
Project description:Tapping panel dryness (TPD) seriously affects the natural rubber (NR) production of Hevea brasiliensis (rubber tree). Several studies have speculated that TPD influences NR biosynthesis in the latex of rubber trees based on the expression changes of NR biosynthesis-related genes. In this study, iTRAQ analysis of latex were carried out to reveal the molecular mechanism of TPD affecting rubber trees NR biosynthesis activity and molecular weight.
2023-05-10 | PXD027892 | Pride
Project description:This study is committed to de novo sequencing and comparative analysis of the transcriptomes of healthy (H) and Tapping panel dryness (TPD)-affected (T) rubber trees to identify the genes and pathways related to the TPD. Total raw reads of 34,632,012 and 35,913,020 bp were obtained from H and T library, respectively using Illumina Hiseq 2000 sequencing technology. De novo assemblies yielded 141,456 and 169,285 contigs, and 96,070 and 112,243 unigenes from H and T library, respectively. Among 66535 genes, 107021 genes were identified as differential expressed genes between H and T library via comparative transcript profiling. A majority of genes involved in natural rubber biosynthesis and jasmonate synthesis with most potential relevance in TPD occurrence were found to be differentially expressed. In TPD-affected trees, the expression of most genes related to the latex biosynthesis and jasmonate synthesis was severely inhibited and it probably the direct cause of the TPD. Our de novo transcriptome data sets provide a significant resource for the discovery of genes related to TPD and improve our understanding the occurrence and maintainace of TPD.
2015-03-25 | GSE67214 | GEO
Project description:ngs2014_07_hevea-hevea_tpd-seq-RNAseq analysis of latex samples from healthy and Tapping Panel Dryness-affected trees.-Identification of Tapping Panel Dryness (TPD)-affected trees in a polyclonal trials grown under standard condition. Trees were tapped since November 2010 every 2 days. Latex yield and TPD occurrence were monitored as well as latex RNA samples were collected twice a year for further analysis. At the end of the experiment, gene expression in latex of healthy and TPD trees were compared.
2018-06-06 | GSE101568 | GEO
Project description:The Russian dandelion Taraxacum koksaghyz synthesizes considerable amounts of high molecular weight rubber in its roots. The cis-1,4-isoprene polymers are stored as rubber particle in the latex, the cytosol of specialized cells, called laticifers. The rubber transferase complex localizes at the particle surface and catalyzes the elongation of the cis-1,4-isoprene chains. The heteromeric complex is composed of an NgBR-like protein and a cis-prenyltransferase. In T. koksaghyz two NgBR homologous proteins, TkCPTL1 and TkCPTL2 were identified. Spatial expression analysis showed that TkCPTL1 is predominantly expressed at a high rate in latex, whereas TkCPTL2 exhibits low mRNA levels in all tissues. We investigated the functions of TkCPTL1&2 in two RNAi approaches. The down-regulation of TkCPTL2 showed no altered phenotype indicating a redundant function of the protein in dandelion. Knockdown of TkCPTL1 led to abolished poly(cis-1,4-isoprene) synthesis indicating that TkCPTL1 is essentially contributing to the formation of the rubber polymer. Simultaneously, levels of triterpenes and inulin in roots were significantly elevated. Analyses of latex from TkCPTL1-RNAi plants yielded a newly appearing latex fraction still containing particles, which seemed to be unaffected in structure compared to native rubber particles. Particle size, phospholipid composition and presence of small rubber particle proteins were determined for this purpose. We could show that the particles encapsulate triterpenes in a phospholipid shell that is stabilized by SRPPs. MS-based comparison of latex proteomes from TkCPTL1-RNAi plants and T. koksaghyz wildtypes enabled evidences for regulation mechanisms in several metabolic pathways associated to rubber biosynthesis.
2022-03-02 | PXD010953 | Pride
Project description:MicroRNAs (miRNAs) are non-coding, short, single-stranded RNAs with essential roles in gene regulation in various organisms including higher plants. In contrast to the vast information on miRNAs from many economically important plants, almost nothing has been reported on the identification or analysis of miRNAs from rubber tree (Hevea brasiliensis L.), the most important natural rubber-producing crop. To identify miRNAs and their target genes in rubber tree, high throughput sequencing combined with a computational approach was performed. Four small RNA libraries were constructed for deep sequencing from mature and young leaves of two rubber tree clones, PB 260 and PB 217, which provide high and low latex yield, respectively. 237 miRNAs belonging to 37 known miRNA families were identified, and northern hybridization validated miRNA expression and revealed developmental stage-dependent and clone-specific expression for some miRNAs. We took advantage of the newly released rubber tree genome assembly as well as the genomic databases from leafy spurge and cassava, two species related to rubber tree, and predicted 15 novel miRNAs.
2012-04-03 | GSE35254 | GEO