organism: Lupinus angustifolius
description: Protocols: Tissue sampling was performed at 10 AM at 1, 5, 9, 16 and 23 days post inoculation. Samples were immediately frozen in liquid nitrogen and stored in -80 °C until RNA extraction. Every sample consisted of 10 mm stem section carrying scarification site (in the middle) with surrounding regions, weighing approximately 90 mg. Experiment was performed in controlled environment (a greenhouse), in temperature regime 21 °C day / 16 °C night and under 14 h photoperiod. Pots (11 cm × 11 cm × 21 cm) with sterilized soil were used. Inoculation was performed 4 weeks after sowing at 10 AM. Lower parts of stems were scarified by lancet (5 cm from root neck) and inoculated with 20 μl of conidia suspension of a given isolate of D. toxica (106 conidia per ml). Inoculation sites were wrapped with Parafilm (Sigma-Aldrich, Poznan, Poland) to minimize evaporation. Control plants were also scarified but no inoculum was applied. After inoculation plants were grown with a relative humidity above 80%.s Every frozen plant sample was divided into two parts (each of 45 mg) and proceeded in parallel. Stem fragments were cut into 5 pieces (about 10 mg each) and homogenized using TissueLyser II (Qiagen) and two stainless steel beads ø 5 mm (Qiagen) in 2 ml tubes (Eppendorf, Hamburg, Germany). RNA isolation was performed using Spectrum Plant Total RNA Kit (Sigma-Aldrich) with some alterations to the standard lysis protocol based on our previous experiences. TruSeq RNA Sample Preparation v2 Guide, Part # 15026495 Rev. F
attributes.tag: ENA first public
attributes.value: 2021-01-12
attributes.tag: ENA last update
attributes.value: 2020-11-25
attributes.tag: External Id
attributes.value: SAMEA7625146
attributes.tag: INSDC center alias
attributes.value: Department of Biometry and Bioinformatics, Institute of Plant Genetics, Polish Academy of Sciences
attributes.tag: INSDC center name
attributes.value: Department of Biometry and Bioinformatics, Institute of Plant Genetics, Polish Academy of Sciences
attributes.tag: INSDC first public
attributes.value: 2021-01-12T04:59:48Z
attributes.tag: INSDC last update
attributes.value: 2020-11-25T15:42:49Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-9814:Sample 54
attributes.tag: age
attributes.value: 29
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: narrow-leaved blue lupine
attributes.tag: cultivar
attributes.value: 75A:258
attributes.tag: disease
attributes.value: normal
attributes.tag: organism part
attributes.value: stem internode
attributes.tag: phenotype
attributes.value: very resistant, Phr1 allele
attributes.tag: replicate
attributes.value: 3
attributes.tag: sample name
attributes.value: E-MTAB-9814:Sample 54