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SRX6756341: GSM4044049: MS_01; Durio zibethinus; RNA-Seq
1 ILLUMINA (Illumina HiSeq 4000) run: 21.3M spots, 6.4G bases, 2.4Gb downloads

Submitted by: NCBI (GEO)
Study: Discovery of Novel Genes and Pathways through Comparative Transcriptomics Approach to Elucidate Genes Expressed in Durian (Durio zibethinus Murr) D24 Varieties Fruit Pulp Development and Ripening.
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In this study, RNA Seq analysis was performed for the comprehensive understanding of the transcriptome of the durian, Durio zibethinus Murr. using Illumina HiSeq Platform. In comparison with the durian reference genome, the studies identified 3,438 novel exons and 1,197 novel loci. The total number of novel transcripts were 280. Ninety-seven novel transcripts did not show any similarity with any sequences available in the Genebank's nucleotide database. Comparison of the top five ranked gene abundances between three differential groups YS/MS, YS/RS and MS/RS showed a difference in their genes and the function. Several key genes involved in sugar metabolism, sulfur production, and ripening process were further studied and visualised using IGV to view changes in their expression profile. 1,4-alpha-glucan-branching enzyme 2-1, chloroplastic/amyloplastic-like, hexokinase-3-like, UDP-sugar pyrophosphorylase-like were detected among the genes responsible for sugar and starch metabolism in durian fruit. The highest expression of genes involved in sugar and starch metabolism pathway was in the mature stage. There is almost similar level of gene expression involved in sugar metabolism in young and ripening stage. Methionine gamma-lyase-like (MGL) and PAP-specific phosphatase HAL2-like genes responsible for sulphur production and the metabolism in durian showed a rapid change in expression (about two times) from the young stage to mature and ripening stage. Interestingly, 1-aminocyclopropane-1-carboxylate oxidase (ACO) genes responsible for ethylene biosynthesis for fruit ripening were showed a significant increase in young stage compared to other stages. Glucose and ribitol dehydrogenase-like were among the top gene abundance identified in the ripening stage, and responsible in carbohydrate metabolism and acquisition of desiccation tolerance. Endochitinase 1-like was the gene in all three stages and responsible in carbohydrate binding site. The categorization of differently expressed transcripts among the three groups to Gene Ontology (GO) and KEGG pathway indicates the highest number of assigned genes to cellular process, metabolic process, cellular metabolic process, cellular nitrogen compound metabolic process; (biological_process), binding, catalytic activity, ion binding (molecular_function) and intracellular, cell part, cell (cellular_component). KEGG pathway mapping was finally carried out to classify the genes into different metabolic pathway. The percentage of the mapped reads to the KEGG pathway was 96.50% (YS/MS), 60.20% (YS/RS) and 35.30% (MS/RS). KEGG pathway distributions of highly expressed genes were mapped to the pathway in a group of nucleotide metabolism, carbohydrate metabolism, lipid metabolism, amino acid metabolism, metabolism of other amino acids, metabolism of cofactors and vitamins, biosynthesis of other secondary metabolites, glycan biosynthesis and metabolism, metabolism of terpenoids and polyketides, and energy metabolism. Comparison of the output of the top 10 highly significant genes between four statistical methods; CLC, CuffDiff, DESeq2, and EdgeR showed the similarity and dissimilarity between the pipelines. The highest similarity of the significant genes was observed when compared using DESeq2 and EdgeR methods. As a whole, the studies conducted successfully revealed several genetic attributes of durian (D24) cultivar. The information also serves as a preliminary genomics expression data which could serve as the foundation in creating a new durian variety that is drought-resistance, and has longer shelf-like and has low-sugar fruits suitable for diabetics and glucose intolerant patients. Overall design: Total of 9 samples; 3 biological replicates collected from three different stages of growth; young, mature and ripening,
Sample: MS_01
SAMN12629377 • SRS5305314 • All experiments • All runs
Library:
Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: RNA extraction kit namely GeneAll RibospinTM Seed/Fruit RNA mini kit was used to isolate the RNA. RNA samples were extracted according to the manufacturer's protocols with the minor modifications to increase the yileds and purity. The assesment of RNA quality was carried out using implen nanodrop spectrophotometer, qubit fluorometer, 1% gel agarose electrophoresis, agilent bioanalyzer and agilent tapestation. Poly-A mRNAs were purified from 1 ug of total RNA using the NEB Next Poly(A) mRNA magnetic isolation module. The first and second strand of cDNA synthesis was performed with purified mRNA using the NEB Next Ultra RNA Library Prep Kit (Ilumina). The index primers 1-9 were used for each of the samples to ensure every samples were having different barcode for pool method of sequencing. To obtain the final library, the cDNA fragments were amplified by PCR as per outlined in the NEB protocol. The purification of PCR reaction were carried out using AmPure XP beads. Nine pre-prepared cDNA libraries were send to NGS service provider (Novogene). The libraries were sequenced with HiSeq Ilumina Platform using 2x 150 bp paired end protocol.
Experiment attributes:
GEO Accession: GSM4044049
Links:
Runs: 1 run, 21.3M spots, 6.4G bases, 2.4Gb
Run# of Spots# of BasesSizePublished
SRR1001869821,308,8086.4G2.4Gb2023-01-25

ID:
8900779

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