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Status |
Public on Sep 01, 2023 |
Title |
Tolerant callus line L3, non exposed to fungal filtrate TCL3 |
Sample type |
SRA |
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Source name |
Callus
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Organism |
Persea americana |
Characteristics |
cultivar: Anaheim tissue: Callus cell line: L3 phenotype: Tolerant cell type: Embryogenic treatment: Non exposed to fungal filtrate
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Treatment protocol |
The virulent CH53 fungal strain of Rosellinia necatrix was cultured on potato dextrose agar (PDA) in darkness at 25 °C. To obtain the fungal filtrate, six 1cm-fragments of PDA with fungal mycelium were incubated on 500ml of PDA liquid medium during 9 days at 25 °C in darkness. This medium was filtrated through 2 layers paper to eliminate hyphae and remains of the fungus; the medium obtained was sterilized through 0.2 µm filtration and frozen until its use.
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Growth protocol |
The embryogenic cultures were maintained on Murashige and Skoog (MS) medium (Murashige & Skoog, 1962) supplemented with 0.1 mg l-1 picloram (MSP medium) and solidified with 6 g l-1 agar in darkness at 25 ± 1 °C and were subculture at 4-week intervals in MSP medium. The pH medium was adjusted to 5.74 before to autoclaving at 0.1 MPa and 121 °C for 15 min.
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Extracted molecule |
total RNA |
Extraction protocol |
For RNA extraction, the embryogenic callus from L3 and AN9 exposed and no exposed to fungal exudates were macerated with liquid nitrogen. And, afterwards, RNA was extracted using ´Spectrum plant RNA kit´ (Sigma Aldrich) following manufacturer’s instructions. Sequencing libraries were generated using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA) according to the manufacturer.
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Library strategy |
RNA-Seq |
Library source |
transcriptomic |
Library selection |
cDNA |
Instrument model |
Illumina NovaSeq 6000 |
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Description |
R09TCL
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Data processing |
Raw data in a FASTQ format were processed using fastp. Raw reads were processed to remove reads containing adapter and poly-N sequences and reads with low quality and were calculated the Q20, Q30 and GC content of the clean data (clean reads). High quality paired-end clean reads were mapped to the reference genome downloaded from genome website browser (NCBI) using HISAT2 software. Differential expression analysis of three biological replicates per condition was performed using DESeq2 R package. Benjamin and Hochberg’s approach were used to adjust P values for controlling the False Discovery Rate (FDR) and select genes with an adjusted Pvalue < 0.05 as differentially expressed genes. The main purpose of this study is to compare and analyse differential expressed genes (DEGs) and differential pathways after growing two different avocado lines (S, Susceptible and T, Tolerant) on media supplemented with Rosellinia necatrix filtrates. A differential expression analysis is made. The input data for differential gene expression analysis is read counts from gene expression level analysis. The differential gene expression analysis contains three steps: read counts normalization; model dependent p-value estimation; FDR value estimation based on multiple hypothesis testing. Assembly: not provided Supplementary files format and content: Differential expression analysis between two conditions/groups (three biological replicates per condition) was performed using DESeq2 R package. DESeq2 provides statistical routines for determining differential expression in digital gene expression data using a model based on the negative binomial distribution. The resulting P values were adjusted using the Benjamini and Hochberg’s approach for controlling the False Discovery Rate (FDR). Genes with an adjusted P value < 0.05 found by DESeq2 were assigned as differentially expressed.
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Submission date |
Mar 27, 2023 |
Last update date |
Sep 01, 2023 |
Contact name |
Clara Pliego |
E-mail(s) |
mclara.pliego@juntadeandalucia.es
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Phone |
0034654564365
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Organization name |
IFAPA
|
Department |
Department of Genomics and Biotechnology
|
Lab |
Biotechnology
|
Street address |
Cortijo de la Cruz
|
City |
Malaga |
State/province |
Malaga |
ZIP/Postal code |
29140 |
Country |
Spain |
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Platform ID |
GPL33284 |
Series (1) |
GSE228295 |
Transcriptome analysis of avocado embryogenic cultures selected for resistance to Rosellinia necatrix exudates |
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Relations |
BioSample |
SAMN33939821 |
SRA |
SRX19785843 |