Sample 29
Lupinus angustifolius
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
ENA first public
2021-01-12
ENA last update
2020-11-25
External Id
SAMEA7625109
INSDC center alias
Department of Biometry and Bioinformatics, Institute of Plant Genetics, Polish Academy of Sciences
INSDC center name
Department of Biometry and Bioinformatics, Institute of Plant Genetics, Polish Academy of Sciences
INSDC first public
2021-01-12T04:59:47Z
INSDC last update
2020-11-25T15:42:48Z
INSDC status
public
Submitter Id
E-MTAB-9814:Sample 29
age
29
broker name
ArrayExpress
common name
narrow-leaved blue lupine
cultivar
Wonga
disease
normal
organism part
stem internode
phenotype
moderately resistant, PhtjR allele
replicate
2
sample name
E-MTAB-9814:Sample 29