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Murashige and skoog ms231 medium

Manufactured by Duchefa Biochemie

Murashige and Skoog MS231 medium is a plant tissue culture medium formulated by Toshio Murashige and Folke Skoog. It provides a balanced nutrient solution for the in vitro growth and development of plant cells, tissues, and organs.

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2 protocols using murashige and skoog ms231 medium

1

Arabidopsis Growth and Genetic Manipulation

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A. thaliana plants were grown in long-day conditions (16 h-day-at 21°C / 8 h night at 18°C cycles, LED tubes Philips 1500 mm SO 20W 840 T8, photon flux density of 120 µmol/s/m 2 at the plant level). Seedlings were grown for 8 days in hydroponic cultures at 23°C with constant light in Murashige and Skoog MS231 medium (Duchefa).
The FMT insertion mutant SALK_056717 was from Columbia ecotype. The line expressing the epitope-tagged ribosomal protein, FLAG-RPL18 (AT3G05590), was from (Zanetti et al., 2005) .
FMT-GFP, GFP-FMT and MS2-GFP lines were obtained by floral dip with pAM557, pAM549 and pAM495 plasmids respectively (Table S6) {Clough, 1998 #958}.
Transient transformation of At and N. benthamiana was performed by infiltration of leaves with a suspension of Agrobacterium tumefaciens harboring different constructs. The constructs allowed the expression of different GFP or split-GFP constructs, the pSu9-RFP mitochondrial marker (Michaud et al., 2014) , the COXIV-GFP mitochondrial marker (Peeters et al., 2000) and the silencing suppressor P19 protein (Michaud et al., 2014) .
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2

Plant Stress Response Protocols

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The insertion mutants SALK_034653 for vdac1 and SALK_127899.41.10.x for vdac3 [5 (link)] were from the Columbia ecotype. Both lines were kindly provided by Dr. Filleur (I2BC, Gif/Yvette, France). vdac1_OEV3 corresponds to a vdac1 mutant transformed with the VDAC3 genomic sequence (from its promoter to the end of its short 3′ UTR, and with a HA tag at the beginning of the coding sequence (CDS) [7 (link)]. Arabidopsis cell cultures were from the Landsberg ecotype [8 (link)].
Plants were grown in long-day conditions (16 h day at 21 °C/8 h night at 18 °C cycles, LED tubes Philips 1500 mm SO 20W 840 T8, photon flux density of 120 mmol/s/m2 at the plant level). Seedling cultures and cell cultures were grown at 23 °C with constant light. Seedlings were grown in Murashige and Skoog MS231 medium (Duchefa Biochemie, Haarlem, The Netherlands) (8 days in hydroponic cultures or 10 days on plates for O2 consumption experiments). Cell cultures were grown in Murashige and Skoog MS256 medium.
Stress conditions were performed on 8-day-old cell cultures. Cold and heat shocks were for 3 h at 4 °C or 37 °C, respectively. NaCl (150 mM) and H2O2 (20 mM) stresses and phosphate starvation were for 24 h at 23 °C. For phosphate starvation studies, media used were MSP01 (with phosphate) and MSP11 (without phosphate) (Caisson Labs, Smithfield, UT, USA).
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