Culturing Physcomitrella patens Mutants

Physcomitrella patens subspecies patens (Hedwig) Bruch & Schimp. WT strain “Gransden 2004”, used for genome sequencing^{32 (link)}, provided the genetic background for the generation of ΔPpSCRM1 mutants (“Gransden 2004”, Freiburg) and ‘Villersexel’ the genetic background for the generation of ΔPpSMF1 mutants (Sheffield), and “Gransden D12” was the background for production of the ΔPpSMF2 mutants. P. patens was grown axenically on BCDAT medium^{33 (link)} supplemented with 1 mM calcium chloride and overlaid with cellophane discs (AA Packaging, UK), in 9 cm Petri dishes sealed with Micropore tape (3M) in Sanyo MLR incubators under continuous light (140 μmol m⁻² s⁻¹) at 25 °C^{34 (link)}. P. patens (Freiburg) was grown in liquid or on solid (12 g/L purified agar (OXOID, Thermo Scientific, Waltham, MA, USA)) supplemented Knop medium^{35 (link),36 (link)} and cultivated at 23 °C under a 16-hour light and 8-hour dark cycle^{37 (link)}. Sporophyte development was induced according to ref^{38 (link)}.

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Chater C.C., Caine R.S., Tomek M., Wallace S., Kamisugi Y., Cuming A.C., Lang D., MacAlister C.A., Casson S., Bergmann D.C., Decker E.L., Frank W., Gray J.E., Fleming A., Reski R, & Beerling D.J. (2016). Origin and function of stomata in the moss Physcomitrella patens. Nature plants, 2, 16179.

Publication 2016

Micropore tape OXOID

Agar Calcium chloride Cellophane Dishes Genetic background Genome Light Physcomitrella Strain

Corresponding Organization : University of Freiburg

Other organizations : Universidad Nacional Autónoma de México, University of Sheffield, Royal College of Veterinary Surgeons, University of Leeds, University of Michigan–Ann Arbor, Stanford University, Ludwig-Maximilians-Universität München

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Variable analysis

independent variables

Genetic background of Physcomitrella patens
Growth medium (BCDAT medium vs. Knop medium)
Light conditions (continuous light vs. 16-hour light/8-hour dark cycle)
Temperature (25°C vs. 23°C)

dependent variables

Physcomitrella patens growth and development
Sporophyte development

control variables

Axenic growth conditions
Cellophane discs used for growth on BCDAT medium
Agar concentration (12 g/L) for solid Knop medium

positive controls

Wild-type 'Gransden 2004' strain used as the genetic background for generation of Δ PpSCRM1 mutants
'Villersexel' used as the genetic background for generation of Δ PpSMF1 mutants
'Gransden D12' used as the genetic background for production of Δ PpSMF2 mutants

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