Arabidopsis thaliana mutants lib (Yu et al., 2013 (link)), coi1-1 (Xie et al., 1998 (link)), and jar1-1 (Staswick et al., 2002 (link)) were described previously. In all experiments, the Columbia-0 (Col-0) was used as WT plants. Meanwhile, coi1-1 homozygous seedlings were screened out via supplying JA in MS medium (Xie et al., 1998 (link)). Arabidopsis seeds were sterilized with 20% bleach, plated on Murashige and Skoog medium (MS; Sigma-Aldrich, United States), cooled at 4°C for 2 days, and then grown in a growth room under a 16-h light/8-h dark (21–23°C) photoperiod.
Murashige and skoog ms medium
Manufactured by Merck Group
Sourced in United States
Murashige and Skoog (MS) medium is a widely used plant cell culture medium formulated by Toshio Murashige and Folke Skoog. It provides a balanced set of essential macro and micronutrients, as well as vitamins, to support the growth and development of plant cells, tissues, and organs in vitro.
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Lab products found in correlation
Fm4 64, by Thermo Fisher Scientific (3 mentions)
Phytagel, by Merck Group (2 mentions)
Tyra23, by Merck Group (1 mentions)
Methyl 1 butylcarbamoyl 2 benzimidazolecarbamate benomyl, by Merck Group (1 mentions)
Indole acetic acid iaa, by Merck Group (1 mentions)
Flg22, by MedChemExpress (1 mentions)
6 benzylaminopurine, by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
Mannitol, by Merck Group (1 mentions)
Hygromycin b, by Merck Group (1 mentions)
Filter paper, by Cytiva (1 mentions)
Agarose, by Merck Group (1 mentions)
Plant agar, by Duchefa Biochemie (1 mentions)
High fidelity dna polymerase, by Thermo Fisher Scientific (1 mentions)
Taq dna polymerase, by Thermo Fisher Scientific (1 mentions)
53 protocols using murashige and skoog ms medium
1
Arabidopsis mutant growth protocol
2
Developmental Transcriptomics of Faba Bean
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V. faba L. (cv. Fiord) plants were grown in environmentally-controlled glasshouse and growth cabinet conditions as previously described (Offler et al., 1997 (link); Farley et al., 2000 (link)). To sample transcripts from cotyledons undergoing epidermal TC development, cotyledons (100–120 mg FW) were removed from pods and either fixed immediately (t = 0-h) in ice-cold ethanol:acetic acid (3:1 v/v) for 1 h, or placed adaxial surface down on filter paper soaked in Murashige and Skoog medium (MS; Sigma Australia) in petri dishes. The Petri dishes were sealed with tape and incubated in darkness at 22°C for 3, 9, or 24 h, then the cotyledons were fixed in ice-cold ethanol:acetic acid as described above. The ethanol-acetic acid fixation was used for all cotyledons to minimize wounding responses associated with handling. Fixed cotyledons were rinsed briefly in distilled water and sheets of adaxial epidermal cells and discs of storage parenchyma were then isolated from each cotyledon as described (Dibley et al., 2009 (link)). For each time-point (0, 3, 9, 24 h) epidermal peels and storage parenchyma disks were collected from a minimum of 10 cotyledons derived from pods harvested from at least three different plants, and were snap frozen in liquid nitrogen and stored at –80°C prior to RNA isolation.
3
Molecular Tools for Plant Signaling Analysis
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LaCl3 (purity >99.99%) was purchased from Aladdin Bio-Chem Technology Co. Ltd (Shanghai, China). FM4-64 was purchased from Thermo Fisher Scientific Co. Ltd (Shanghai, China). DPI, TyrA23, Murashige and Skoog medium (MS), methyl 1-(butylcarbamoyl)-2-benzimidazolecarbamate (Benomyl), 6-benzylaminopurine, indole acetic acid (IAA), and JA were purchased from Sigma-Aldrich Co. Ltd (Shanghai, China). Flg22 was purchased from MedChemExpress Co. Ltd (Shanghai, China). Anti-plant actin mouse monoclonal antibody (A01050) was purchased from Abbkine Scientific Co. Ltd (Wuhan, China). Goat anti-mouse IgG H&L [horseradish peroxidase (HRP)] (ab205719) and HRP anti-GFP antibody (ab6663) were purchased from Abcam Co. Ltd (Shanghai, China). Other chemicals used in this study were analytical reagents and were purchased from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China).
4
Gerbera and Arabidopsis Growth Protocols
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Gerbera hybrida “Shenzhen No. 5” seedlings were grown under standard greenhouse conditions at 26/18°C (day/night temperature) and a relative humidity of 65–80%. Ray petals at different developmental stages (Meng and Wang, 2004 (link)), and other tissues and organs were sampled for the following experiments.
Arabidopsis thaliana seeds (Col-0) were surface sterilized, plated on Murashige and Skoog medium (MS) (Sigma–Aldrich), and imbibed in darkness at 4°C for 3 days for vernalization. The plates were transferred to a growth room (21–22°C, 60% relative humidity) under long-day conditions (16 h light/8 h dark) for 7 days, then seedlings were transplanted into pots containing peat:vermiculite (3:1) and grown under the same conditions.
Arabidopsis thaliana seeds (Col-0) were surface sterilized, plated on Murashige and Skoog medium (MS) (Sigma–Aldrich), and imbibed in darkness at 4°C for 3 days for vernalization. The plates were transferred to a growth room (21–22°C, 60% relative humidity) under long-day conditions (16 h light/8 h dark) for 7 days, then seedlings were transplanted into pots containing peat:vermiculite (3:1) and grown under the same conditions.
5
Investigating Cotton Fiber Development
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Gossypium hirsutum acc. Texas Marker-1 (TM-1), the fuzzless or naked seed mutant n2NSM, and the fuzzless–lintless mutant XZ142FLM were used to investigate fiber development. All plants were grown at the Dangtu Breeding Station (DBS/NAU) with normal field practices, and in the glasshouses of Nanjing Agriculture University. Arabidopsis seeds were sterilized with 20% bleach, plated on Murashige and Skoog medium (MS; Sigma-Aldrich), chilled at 4 °C for 3 d, and transferred to a growth room with a 16 h (22–24 °C)/8 h (19 °C) light/dark photoperiod. Nicotiana benthamiana was grown under a 16 h (28 °C)/8 h (22 °C) light/dark photoperiod.
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Gossypium hirsutum acc. Texas Marker-1 (TM-1), the fuzzless or naked seed mutant n2NSM, and the fuzzless–lintless mutant XZ142FLM were used to investigate fiber development. All plants were grown at the Dangtu Breeding Station (DBS/NAU) with normal field practices, and in the glasshouses of Nanjing Agriculture University. Arabidopsis seeds were sterilized with 20% bleach, plated on Murashige and Skoog medium (MS; Sigma-Aldrich), chilled at 4 °C for 3 d, and transferred to a growth room with a 16 h (22–24 °C)/8 h (19 °C) light/dark photoperiod. Nicotiana benthamiana was grown under a 16 h (28 °C)/8 h (22 °C) light/dark photoperiod.
6
Nematode Infection Assay in Transgenic Arabidopsis
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MiMIF-2 transgenic A. thaliana lines were previously described (Zhao et al., 2019 (link)). Wild-type (WT) A. thaliana (Col-0) was used as a control. Surface-sterilized A. thaliana seeds were placed on Murashige and Skoog Medium (MS) (Sigma, St. Louis, MO, USA) and were incubated at 22°C with a 12-h photoperiod. Seedlings of 15 days old plants were transplanted into soil and were grown in the greenhouse at 22°C and 65% relative humidity with a 16-h light/8-h dark photoperiod.
Tomato plants (Solanum lycopersicum var. “Baiguo”) were used to reproduce M. incognita nematode culture. Infective pre-parasitic second-stage juveniles (pre-J2s) were collected after egg hatching (Zhao et al., 2019 (link)).
Tomato plants (Solanum lycopersicum var. “Baiguo”) were used to reproduce M. incognita nematode culture. Infective pre-parasitic second-stage juveniles (pre-J2s) were collected after egg hatching (Zhao et al., 2019 (link)).
7
Hyptis suaveolens Seed Germination Protocol
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Plants and seeds of Hyptis suaveolens were collected in Merida,
Yucatan, Mexico, in 2006 and authenticated by German Carnevalli, Scientific
Research Center of Yucatan Herbarium. A specimen of the plant was deposited
(voucher number CICY 7086). The seeds were carefully washed under running tap
water for 10 min and surface-sterilized by immersing in ethanol at 70% (v/v) for
1 min and in sodium hypochlorite at 1.5% (v/v) for 5 min. The sterilized seeds
were immediately rinsed three times with sterile distilled water for 5 min.
Subsequently, the seeds were placed in glass vessels containing Murashige and
Skoog medium (MS, Sigma) [18 (link)], amended with sucrose 3% (w/v), and 2.5 g L-1water-phytagel for germination. The plantlets were maintained until they were 5
cm in length in a growth chamber at 24 ± 2°C with a photoperiod of 16/8 h
light/darkness. Light sources were cool white lamps of 40 watts.
Yucatan, Mexico, in 2006 and authenticated by German Carnevalli, Scientific
Research Center of Yucatan Herbarium. A specimen of the plant was deposited
(voucher number CICY 7086). The seeds were carefully washed under running tap
water for 10 min and surface-sterilized by immersing in ethanol at 70% (v/v) for
1 min and in sodium hypochlorite at 1.5% (v/v) for 5 min. The sterilized seeds
were immediately rinsed three times with sterile distilled water for 5 min.
Subsequently, the seeds were placed in glass vessels containing Murashige and
Skoog medium (MS, Sigma) [18 (link)], amended with sucrose 3% (w/v), and 2.5 g L-1water-phytagel for germination. The plantlets were maintained until they were 5
cm in length in a growth chamber at 24 ± 2°C with a photoperiod of 16/8 h
light/darkness. Light sources were cool white lamps of 40 watts.
8
Arabidopsis Growth under Bacterial Inoculation
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Arabidopsis thaliana Col-0 seeds were surface sterilized with 70% ethanol for 1 min, followed by 1% commercial chlorine bleach and 0.01% Tween 20 solution for 7 min, and then washed three times with sterile distilled water. The sterilized seeds were then sown on 1% agar plates containing ½ Murashige and Skoog medium (MS) (Sigma-Aldrich) (Murashige and Skoog, 1962 (link)). Some plates were inoculated with bacteria. Eight seeds were sown in each plate and three plates were used for each treatment. Plates were vertically placed in the incubator at 24°C with a photoperiod of 12 h light and 12 h of dark. Several growth parameters were measured 10 days after sowing. For pot experiments, seeds were sterilized and immersed into bacterial suspension for 10 s, then sown into pots with sterile soils maintained at the same environmental conditions as described above for 2 months. Plants were watered with sterile water twice per week. Ten seeds were sown in each pot and three pots were used for each treatment. The same experiment was repeated three times.
9
Arabidopsis and Populus Growth Conditions
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Arabidopsis thaliana, ecotype Columbia (Col‐0), was used in both wild‐type and transgenic plant experiments. Arabidopsis were grown in soil in a growth room (14 h light; light intensity, 150 μmol/m2/s) at 23 °C or on half‐strength Murashige and Skoog medium (MS, Sigma‐Aldrich) containing 2% sucrose with appropriate antibiotics for screening. Hybrid poplars (Populus alba × P. tremular var. glandulosa, clone BH) were used as both wild‐type controls and transgenic plants in this study. The plants were acclimated in soil and grown in controlled conditions in a growth room (16 h light; light intensity, 150 μmol/m2/s; 24 °C).
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Arabidopsis thaliana, ecotype Columbia (Col‐0), was used in both wild‐type and transgenic plant experiments. Arabidopsis were grown in soil in a growth room (14 h light; light intensity, 150 μmol/m2/s) at 23 °C or on half‐strength Murashige and Skoog medium (MS, Sigma‐Aldrich) containing 2% sucrose with appropriate antibiotics for screening. Hybrid poplars (Populus alba × P. tremular var. glandulosa, clone BH) were used as both wild‐type controls and transgenic plants in this study. The plants were acclimated in soil and grown in controlled conditions in a growth room (16 h light; light intensity, 150 μmol/m2/s; 24 °C).
10
Bacterial colonization of Arabidopsis
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The strain Pseudomonas japonica S3R2.1 was chromosomally tagged with a gene coding for a red fluorescent protein (DsRed) using a mini-Tn7 transposon system by conjugation procedure using E. coli Lam#5as donor (Lambertsen et al., 2004) . The transconjugant strain P. japonica S3R2.1_DR showed resistance to rifampicin and kanamycin conferred by resistance genes harbored by the transposon system and exhibited red fluorescence under epifluorescent microscopy. P. japonicaS3R2.1_DR was used to bacterize Arabidopsis thaliana plantlets as detailed by Mapelli et al. (2013) . Briefly, cells of P. japonica S3R2.1_DR harvested in exponential growth phase were supplemented to A. thaliana by dipping for a short time (1 to 4 hours) the axenic roots in Murashige and Skoog medium (MS, SIGMA, Italy) containing 10 8 cells/ml. Plants were then transferred on sterile MS medium and incubated for 14 days in growth chamber (temperature 25°C, 55% humidity). Plant leaves were randomly sampled by cutting, washed with sterile water, crushed and re-suspended in sterile saline solution (NaCl 0.9%) for Colony-Forming Unit (CFU) determination on TSA medium supplemented by rifampicin (0.1 mg/ml) and kanamycin (0.05 mg/ml). The red fluorescence of randomly chosen colonies was verified by cell observation under epifluorescence microscopy.
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