F1 node 3
Lathyrus oleraceus
Protocols: Whole stipules were harvested from leaflets of individual plants and snap-frozen in liquid nitrogen. Each sample was crushed into a fine powder by pestle and mortar. Samples were then weighed on a microgram scale to 1 mg of whole stipule tissue per extraction.  P. sativum cultivars (supplied by the Australian Grains Genebank and the John Innes Centre) were grown within a controlled glasshouse environment at an average ambient temperature of 22°C and subjected to long-day light cycles (8 h dark, 16 h light). The soil medium macro/micro nutrients and other constituents consisted of 0.5 L Vermiculite, 0.5 L Perlite, 35 g Macracote Coloniser Plus 4-month slow release fertiliser (15N : 3P : 9K), 30 g Nitrogen slow-release fertiliser (40N : 0P : 0K), 25 g Water holding granules, 15 g Trace elements (6Mg : 6.5FE :5.4S : 1.5Mn : 0.4Zn : 0.14B : 0.07Mo), and 5 g Garden lime per 30 L of soil. Total RNA was then isolated using Invitrogen TRIzol reagent (ThermoFisher Scientific) using the manufacturer's protocol. Minor modifications to the protocol included an extended centrifugation for 15 minutes, as some RNA precipitate was still suspended in solution for some samples. RNA precipitate pellets were washed twice rather than once, as it was found this removed further impurities and improved the overall quality of the RNA. Centrifugation between washes was also extended to 10 minutes, to ensure the RNA pellet was anchored to the bottom of the Eppendorf tube during wash steps. Total RNA pellets for each sample were then re-suspended in RNase-free water, and subsequently quality-checked and quantified utilising the NanoDropTM spectrophotometer.  RNA-seq libraries were prepared with the Illumina Truseq Stranded v2 kit, according to manufacturer's instructions
ENA first public
2025-03-01
INSDC center name
WEHI
INSDC status
public
Submitter Id
E-MTAB-14845:F1 node 3
age
6 weeks post emergence
broker name
ArrayExpress
collection date
not collected
common name
garden pea
cultivar
Sutton's Hundredfold wt x Sutton's Early Giant rogue
developmental stage
third leaflet
genotype
rogue
geographic location (country and/or sea)
not collected
isolate
not applicable
organism part
stipule
sample name
E-MTAB-14845:F1 node 3
scientific_name
Pisum sativum