organism: Arabidopsis thaliana
description: Protocols: Plant material was harvested and frozen into liquid nitrogen after 0h, 30min, 1h, 3h and 24h of infestation. Arabidopsis thaliana Col-0 accession was used. Seeds were planted and incubated 5 days at 4ºC in peat moss and vermiculite (3:2 V/V). Plants were then grown in growth chambers (Sanyo MLR-350-H) under control conditions (23ºC±1ºC, >70% relative humidity and a 16 h/8 h day/night photoperiod). A colony of Tetranychus urticae, London strain (Acari: Tetranychidae) provided by Dr. Miodrag Grbic (UWO, Canada), was reared on beans (Phaseolus vulgaris) and maintained in growth chambers (Sanyo MLR-350-H) at 25ºC±1ºC, >70% relative humidity and a 16h/8h day/night photoperiod. Mites were synchronized and four-week-old plants were infested with 20 T. urticae female adults per plant. They were carefully transferred with a brush to the leaf surface. Total RNA was isolated from frozen rosettes and purified by using the RNeasy Qiagen Mini Plant Kit (74904 Qiagen), including the on-column DNA I (79254, Qiagen) digestion recommended by the manufacturer. RNA amount and quality were tested in a Nanodrop ND-1000. Using poly-T oligo-attached magnetic beads, mRNAs were purified from the total RNA. Then, the mRNAs were fragmented and cDNA was synthesized using random hexamer-primers, DNA polymerase I and RNase H. The double-stranded cDNAs were purified with magnetic beads and ligated to adaptors for Illumina sequencing. The quality and quantity of the library was verified using an Agilent 2100 Bioanalyzer and an ABI StepOnePlus Real-Time PCR system, respectively.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2021-03-23T08:57:00Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2020-09-25T14:41:14Z
attributes.tag: External Id
attributes.value: SAMEA7339845
attributes.tag: INSDC center name
attributes.value: Centro de Biotecnologia y Genomica de Plantas, Universidad Politecnica de Madrid (UPM) - Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (INIA)
attributes.tag: INSDC first public
attributes.value: 2021-03-23T08:57:00Z
attributes.tag: INSDC last update
attributes.value: 2020-09-25T14:41:14Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-9448:Tur_3h 2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: thale cress
attributes.tag: developmental stage
attributes.value: seedling development stage
attributes.tag: ecotype
attributes.value: 4
attributes.tag: ecotype
attributes.value: Col-0
attributes.tag: infect
attributes.value: Tetranychus urticae
attributes.tag: organism part
attributes.value: rosette
attributes.tag: sample name
attributes.value: E-MTAB-9448:Tur_3h 2
attributes.tag: scientific_name
attributes.value: Arabidopsis thaliana