Seedlings were transferred on solid MS medium with the indicated chemicals: propidium iodide (PI, 10 μM, Sigma-Aldrich or Thermofisher), hydroxyurea (HU, final concentration 5 mM, Sigma-Aldrich) for 24 hours, gibberellic acid (GA, final concentration 10 μM, Sigma-Aldrich) for 1 hour before ablation, paclobutrazol (PAC, final concentration 2 or 10 μM as indicated, Sigma-Aldrich) for 1 hour before ablation, epibrassinolide (EBL, Sigma Aldrich, final concentration 1 μM) for 1 hour before ablation, dexamethasone (DEX, Sigma Aldrich, final concentration 5 μM) for 1 hour before ablation, 1-Naphthylacetic acid f (NAA, Sigma Aldrich, final concentration 1 μM) or 1 hour before ablation, 6-Benzylaminopurine (BAP, Sigma Aldrich, final concentration 50 nM) for 1 hour before ablation.
Gibberellic acid ga
Manufactured by Merck Group
Sourced in United States
Gibberellic acid (GA) is a naturally occurring plant hormone that is widely used in agricultural and horticultural applications. It is a member of the gibberellin family of plant hormones, which play a crucial role in regulating plant growth and development. GA is extracted from cultures of the fungus Gibberella fujikuroi and is available as a laboratory product for research and commercial use.
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Lab products found in correlation
Salicylic acid, by Merck Group (2 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
6 benzylaminopurine bap, by Merck Group (1 mentions)
Paclobutrazol pac, by Merck Group (1 mentions)
Epibrassinolide ebl, by Merck Group (1 mentions)
Hydroxyurea hu, by Merck Group (1 mentions)
Indole 3 acetic acid iaa, by Merck Group (1 mentions)
Methyl jasmonate meja, by Merck Group (1 mentions)
Phytocon culture vessels, by PhytoTechnology Laboratories (1 mentions)
Abscisic acid aba, by Merck Group (1 mentions)
5 protocols using gibberellic acid ga
1
Modulation of Seedling Development
2
Phytohormone Treatments on Seedlings
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Seeds of 30DPH were soaked in 70% (v/v) ethanol for 1 min followed by washing in 2.5% (v/v) sodium hypochlorite solution containing 0.1% (v/v) Tween 20 for 15 min and rinsed thoroughly with sterile distilled water. Surface sterilized seeds were grown in PhytoCon culture vessels (Phytotechnology Laboratories, Overland Park, KS, USA) containing half strength Murashige and Skoog (MS) medium for 14 days. Seedlings were kept in sucrose free liquid half strength MS medium for 24 h. Seedlings of 15DPG (days post germination) were transferred to PhytoCon culture vessels (Phytotechnology Laboratories) containing liquid half strength MS supplemented with 100 µM abscisic acid (ABA, Sigma, St. Louis, MO, USA), 50 µM brassinolide (Bra, Sigma), 50 µM gibberellic acid (GA, Sigma), 50 µM indole-3-acetic acid (IAA, Sigma), 100 µM methyl jasmonate (MeJa, Sigma), 100 µM salicylic acid (SA, Sigma), 100 µM Zeatin (Zea, Sigma) and incubated for 6 h. Leaves from a total of 72 samples from seven treatments in three biological replicates including one untreated control of three genotypes were harvested and immediately frozen as mentioned in the earlier section.
3
Maize Seed Disinfection and Hormonal Treatments
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Zea mays var. VS535 seeds were disinfected with 2% (v/v) household bleach for two min with constant stirring. Hypochlorite was removed from seeds by washing extensively with sterile deionized water. After disinfection, seeds were shaken for 1.5 h at 150 rpm and 30 °C [60 (link)] in 1 mL/seed of sterile deionized water (control), 1 mL/seed of 20,000 F. verticilloides MY3 conidia/seed (infected), 1 mL/seed of 0.5 mM salicylic acid (SA) (Sigma, St. Louis, MO, USA), 1 mL/seed of 0.05 mM methyl jasmonate (MeJa) (Aldrich, St. Louis, MO, USA) or 1 mL/seed of 0.14 mM gibberellic acid (GA) (Sigma, St. Louis, MO, USA). Then, seeds were sown along with their inoculation water or hormone solution on wet filter paper and germinated in the dark and 30 °C for 1, 2, 3, and 6 days, for control and infected conditions, and only for 3 days for hormone treatments. Each experiment was carried out three times, independently.
4
Cotton Ovule Culture for Reproductive Studies
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Cotton (G. hirsutum) was grown in soil in a greenhouse between November 2021 and February 2022 at the Weizmann Institute of Science greenhouse facilities. Flowers were harvested 2 days post anthesis (2 dpa), and the ovary was sterilized in a solution of NaOCl (6%) for 2 min at room temperature. The fertilized ovules were aseptically removed and placed floating onto sterile Beasley and Ting (BT) medium (15 mL) supplemented with gibberellic acid (GA, 5 μM, Sigma, Germany), indoleacetic acid (IAA, 0.5 μM, Germany), and additives (see section “ovule feeding experiments ”). The cultures were kept in the dark at 30°C and 5% CO2 for 20 days (standard conditions).33 (link),34 (link)
5
Comprehensive Fucus Stress Protocol
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Fucus distichus (Fucales, Phaeophyceae) individuals were collected from their natural rockpool habitat at the University of California Kenneth S. ST1). Three biological replicates were obtained for each treatment at two timepoints: 3 hours and 3 days after treatment start, resulting in 90 samples in total. A summary of the treatments may be found in Table S1.
The chemical treatments were: control ASW with nutrients (PES), nutrient-deficient ASW, 0.2µg/L imidacloprid (Marathon 1%, OHP, Inc., USA), 50µM indole-3-acetic acid (IAA;
Cat#102037, MP Biochemicals, Irvine, CA.), 50µM gibberellic acid (GA; Cat#G7645, Sigma). An equal volume of absolute ethanol was used as a control for the GA and IAA treatment. In addition, a saline shock was performed using a hypersaline solution (2x ASW) and a hyposaline solution (0.5x ASW). The desiccation treatment was affected by placing the algal segments onto dry Petri dishes, after blotting gently, under the same environmental conditions as in other treatments. A mechanical wounding treatment, to simulate the effect of grazing, was performed by damaging the algal segments with a razor blade in several places along the thallus. Alteration of light and temperature was achieved as follows: a portion of samples was cultured under modified light conditions (120 μmol m -2 s -1 and complete darkness) or two non-standard temperatures (8⁰C and 22⁰C).
The chemical treatments were: control ASW with nutrients (PES), nutrient-deficient ASW, 0.2µg/L imidacloprid (Marathon 1%, OHP, Inc., USA), 50µM indole-3-acetic acid (IAA;
Cat#102037, MP Biochemicals, Irvine, CA.), 50µM gibberellic acid (GA; Cat#G7645, Sigma). An equal volume of absolute ethanol was used as a control for the GA and IAA treatment. In addition, a saline shock was performed using a hypersaline solution (2x ASW) and a hyposaline solution (0.5x ASW). The desiccation treatment was affected by placing the algal segments onto dry Petri dishes, after blotting gently, under the same environmental conditions as in other treatments. A mechanical wounding treatment, to simulate the effect of grazing, was performed by damaging the algal segments with a razor blade in several places along the thallus. Alteration of light and temperature was achieved as follows: a portion of samples was cultured under modified light conditions (120 μmol m -2 s -1 and complete darkness) or two non-standard temperatures (8⁰C and 22⁰C).
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