Illumina HiSeq 2000 sequencing; Unravelling plant-density genetic architecture in canola
Unravelling plant-density genetic architecture in canola
Greenhouse experimentation was performed with Lynx and Monty genotypes cultivated under simulated shade modelling contrasting plant densities. Seeds were initially sown on agar plates, and transferred to 4L pots in soil:sand:perlite 2:1:1 mix after 9 days (2-3 true leaves stage). They were further grown in greenhouse for 15 days at water field capacity and optimal conditions. Then, plants were separated in two groups: shade and control treatments. The shade treatment was achieved with 50 cm high green filters placed at both sides of pots. Treatment lasted 8 days, along with corresponding control plants with no screen, after which leaves material for RNA extraction were recovered. Samples comprising pooled leaves from at least 6 independent plants were harvested at midday, isolating total RNA from independent triplicated biological samples. Greenhouse growth of Brassica napus plants Total RNA was extracted with the Spectrum Plant Total RNA Kit (Sigma-Aldrich). Non-strand-specific libraries were prepared with the TruSeq mRNA Sample Prep Kit (Illumina), following providerâ€™s instructions.