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SRX19821390: GSM7124365: B728a_P1; Pseudomonas syringae pv. syringae; RNA-Seq
1 ILLUMINA (NextSeq 500) run: 6.1M spots, 615.2M bases, 200.1Mb downloads

External Id: GSM7124365_r1
Submitted by: University of Arizona
Study: Dual-transcriptomic analysis of Nicotiana benthamiana and two compatible Pseudomonas strains
show Abstracthide Abstract
We sought to compare and contrast plant host and bacterial transcriptional changes during compatible infections that cause disease (albeit within different symptoms). We investigated the infection by the two Pseudomonas syringae sensu lato strains P. syringae pv. syringae B728a (Psy) and P. amygdali pv. tabaci 11528 (Pta) of Nicotiana benthamiana at an early time point post inoculation to understand how a plant host responds to two related bacteria with different infection strategies. Plant and bacterial transcriptomes were analyzed prior to and five hours post inoculation. Overall design: RNA was taken from six Nicotiana benthamiana plants before and five hours after syringe inoculation with Psy or Pta for assessing the plant transcriptome. For the bacterial transcriptomes, bacteria growing on King's B medium and from the leaf intercellular wash fluid five hours post inoculation were used for RNA isolation. For each organism and condition there are 3 replicates. To determine whether bacterial transcriptomes were impacted by pelleting before RNA collection, the Psy B728a has a pelleted (P) and not-pelleted (N) set of samples for the in vitro timepoint.
Sample: B728a_P1
SAMN33988076 • SRS17181663 • All experiments • All runs
Library:
Name: GSM7124365
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Frozen samples were homogenized in 2mL homogenization tubes with high-density zirconium beads (Glen Mills) using a Geno/Grinder (SPEX SamplePrep) for 1 min at 1,750 Hz with a LN2 chilled sample holder. RNA was extracted using the DirectZol RNA miniprep kit (Zymo Research). RNA samples were DNAse treated with the Turbo DNAse (Thermo-Fisher) followed by cleanup with the Monarch RNA Cleanup Kit (NEB). Reagents were used according to the manufacturer's recommendations. Plant mRNA sequencing libraries were prepared with the Illumina-compatible KAPA Stranded mRNA-seq Kit (Roche) by the Georgia Genomics and Bioinformatics Core (GGBC). The Psy RNA samples were rRNA depleted using the RiboMinus Bacterial kit (Thermo Fisher) and libraries were prepared with the Illumina-compatible KAPA Stranded RNA-seq Kit (Roche) by the GGBC. The Pta RNA samples were rRNA depleted using both RiboZero Plant and RiboZero Bacteria (Illumina) in combination and RNA sequencing libraries were prepared in-house using the TruSeq Stranded Total RNA kit (Illumina). Library prep kits were used according to the manufacturer's recommendations.
Runs: 1 run, 6.1M spots, 615.2M bases, 200.1Mb
Run# of Spots# of BasesSizePublished
SRR240186576,095,894615.2M200.1Mb2023-09-01

ID:
27168587

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