organism: Medicago truncatula
description: Protocols: M. truncatula lines A17 and skl were treated, grown and inoculated according to (Anderson et al., 2010). R. solani AG8 (isolate WAC10335) was prepared according to (Lichtenzveig et al., 2006). Briefly, surface sterilised M. truncatula seeds were germinated on moist filter paper in the dark at 4 °C for two days, then one day at 24 °C. Seedlings were added to pots half filled with vermiculite also containing 5 ml of 2.2mg/ml dry weight equivalent suspension of homogenised fungal mycelium. The seedlings were covered with fine vermiculite and transferred to growth rooms at a constant temperature of 24 °C with 16/8 light/dark photoperiod at a light intensity of 200 mmol m-2 s-1. Mock treated plants were treated the same with the addition of 5 ml sterile water rather than the 5 ml of R. solani suspension. Inoculated plants were harvested at two and seven days after planting into pre-inoculated pots with corresponding mock samples taken at the same time. Root and above ground tissue was collected separately, immediately frozen in liquid nitrogen and stored at -80 °C until RNA extraction. The remaining plants were scored for survival and harvested at 21 days after inoculation to confirm the infection had progressed as previously described (Anderson et al., 2013). Frozen root tissue was ground in the presence of liquid nitrogen and total RNA extracted using Trizol reagent (Sigma) according to manufacturer's instructions. Libraries were prepared using the illumina TruSeq Stranded mRNA Library Prep Kit according to manufacturer's instructions.
attributes.tag: ENA checklist
attributes.value: ERC000011
attributes.tag: INSDC center name
attributes.value: CSIRO Agriculture and Food
attributes.tag: INSDC first public
attributes.value: 2017-06-06T17:01:35Z
attributes.tag: INSDC last update
attributes.value: 2017-02-13T18:58:55Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: SRA accession
attributes.value: ERS1556020
attributes.tag: Sample Name
attributes.value: ERS1556020
attributes.tag: alias
attributes.value: E-MTAB-5473:skl R. solani inoculated 2 dpi Rep1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: ecotype
attributes.value: sickle mutant
attributes.tag: organism
attributes.value: Medicago truncatula
attributes.tag: organism part
attributes.value: root
attributes.tag: title
attributes.value: skl R. solani inoculated 2 dpi Rep1