This companion website to Barron et al. (2020) provides the workflow integrating protocols, data and reproducible workflow associated to this research. A map showing the locations where seeds from mother plants were collected by our colleagues at the US Forest Service is available here.
Overall, this study is aiming at developing an in vitro method of propagation using growth regulators for Basin Big sagebrush (Artemisia tridentata subsp. tridentata: 2n=2x=18) to support genome sequencing and GxE research.
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Basin Big sagebrush (Artemisia tridentata subsp. tridentata) is the keystone species of the sagebrush steppe, a widespread ecosystem of western North America threatened by climate change. This study aims at developing an in vitro method of propagation for this taxon using growth regulators to support genome sequencing and genotype-by-environment research underpinning drought research. Seedlings were generated from two diploid mother plants (2n=2x=18) collected in environments reflecting contrasting precipitation regimes. Effects of IBA and NAA (at different concentrations) on rooting of shoot tips were compared to a control treatment based on 45 individuals. Number of roots per shoot tip were quantified after 15 days and used to perform statistical (ANOVA) and clustering analyses. Rooted shoot tips were transferred into growing media to assess whether plantlets can be maintained in vitro. Results suggested that A. tridentata is an out-bred species as shown by the significant individual effect on rooting, survival and growth responses. IBA has been shown to significantly promote rooting of shoot tips, especially in the case of the top three performers identified by ANOVA analyses. These individuals have high survival and growth rates, making them suitable candidates for generating biomass for genome sequencing and producing clones for genotype-by-environment research.
The figure below provides an overview of the experimental design applied in this study.