Illumina HiSeq 2500 paired end sequencing; Temperature dependent sex-biased gene expression in the gametophytes of the kelp Saccharina latissima
Temperature dependent sex-biased gene expression in the gametophytes of the kelp Saccharina latissima
Gametophytes from uniparental stock cultures (AWI seaweed collection) of S. latissima (AWI culture Number - males 3094, females 3096) from Helgoland, Germany were cultivated in glass flasks under red light (16hL:8hD) at 12°C in an environmentally controlled room. Seawater enriched with an adapted Provasoli medium without the addition of iron was replaced weekly. These conditions prevented the onset of gametogenesis (Lüning and Dring, 1975; Motomura and Sakai, 1981; Lewis et al., 2013), which was checked under the microscope regularly.  To eliminate the confounding factor of red irradiance in our transcriptomic analysis, red light was replaced by white light with a photon fluence rate of 10 - 15 µmol photons m-2 s -1 (16hL:8hD) provided by fluorescent lamps OSRAM L 18W for one week prior to and during the experiment. These conditions retained our gametophytes' cultures at a vegetative stage. At the start of the experiment 3.10 +/- 0.10 g of fresh weight of female gametophyte clumps and 2.20 +/- 0.10 g of male gametophytes clumps were transferred separately to plastic petri dishes (n=4).  Vegetative gametophytes were isolated from spores of fertile sporophyte collected at Helgoland, Germany according to the protocol of Bartsch I. (2018). Derivation of clonal stock cultures and hybridization of kelps. In Protocols for Macroalgae Research (pp. 61-78): CRC Press Taylor and Francis Group Total RNA extraction was conducted using the method described in Heinrich et al. 2012. Heinrich S, Frickenhaus S, Glöckner G, Valentin K (2012a) A comprehensive cDNA library of light- and temperature-stressed Saccharina latissima (Phaeophyceae). European Journal of Phycology, 47, 83-94.https://www.tandfonline.com/doi/abs/10.1080/09670262.2012.660639 cDNA libraries were prepared with an Illumina TruSeq RNA Library Prep Kit according to the manufacturer protocol.