Fetal bovine serum (FBS), Dulbecco’s modified Eagle’s medium (DMEM), 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), phosphate buffered saline (PBS), antibiotic solution and other chemicals were purchased from Hi-media Limited (Mumbai, India). silica gel (# 100-200 and # 230-400) from Loba Chemie (Mumbai, India), Diaion HP-20® resin from Supelco analytical (USA), Sephadex LH-20 from Amersham Pharmacia Biotech and GE Healthcare (USA), Polyamide 6 and RP18 column from Sigma Chemical Co. (St. Louis, MO, USA) were used for column chromatography. Mouse and rat TNF-α and IL-1ß ELISA kits were obtained from Krishgen Biosystems (Mumbai, India). Thin layer chromatography (TLC) plates pre-coated with silica gel 60 F254 thickness 0.2 mm and solvents (laboratory grade) were received from Merck (Darmstadt, Germany). Dexamethasone, Lipopolysaccharide (Escherichia coli 026:B6) (LPS), dimethyl sulfoxide (DMSO), curcumin, L-NG-Nitroarginine methyl ester (L-NAME), modified Griess reagent, L-mimosine, jasmonic acid and crocin were obtained from Sigma Chemical Co. (St. Louis, MO, USA). For in vivo study, L-mimosine, jasmonic acid and crocin were purchased from Sigma Chemical Co. (St. Louis, MO, USA) and crocetin was prepared by ester hydrolysis of crocin.
Jasmonic acid
Manufactured by Merck Group
Sourced in United States, Germany
Jasmonic acid is a naturally occurring plant hormone that plays a crucial role in plant growth and development. It is a carboxylic acid with the chemical formula C₁₂H₁₈O₃. Jasmonic acid functions as a signaling molecule, regulating various physiological processes in plants, including defense responses, stress tolerance, and reproductive development.
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54 protocols using jasmonic acid
1
Anti-inflammatory Agents Screening Protocol
2
Comprehensive Analysis of Bioactive Compounds
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Jasmonic acid, yeast extract, ferrozine (3-(2-pyridyl)-5,6-bis-(4-phenyl-sulfonic acid)-1,2,4-triazine), 2,20-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), α-amylase, pancreatin, pepsin, lipase, α-glucosidase, bile extract, lipoxygenase, linoleic acid, angiotensin converting enzyme, o-phthalaldehyde, p-nitrophenyl acetate, dimethyl sulfoxide, 3,5-dinitrosalicylic acid, p-nitrophenol, protocatechuic acid, syringic acid, vanillic acid, sinapic acid, salicylic acid, caffeic acid, and hydroxybenzoic acid, cyclohexamide, resazurin were purchased from Sigma-Aldrich (Poznan, Poland). The COX Activity Assay kit was purchased from Cayman Chemical company (Ann Arbor, MI, USA). Penicillin and streptomycin were purchased from Life Technologies, Warsaw, Poland. Mueller–Hinton broth, and Mueller–Hinton agar were also obtained (Biomaxima, Lublin, Poland). All other chemicals were of analytical grade.
3
HPLC-MS Chemical Standards Protocol
4
Analytical Characterization of Nanoparticles and Phytohormones
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LC-MS grade water, acetonitrile (ACN), ethanol and isopropanol were purchased from Fisher Scientific (NJ, USA). Nanoparticles of TiO2, ZnO, bismuth cobalt zinc oxide (Bi2O3)0.07(CoO)0.03(ZnO)0.9 (<100 nm BET or <50 nm XRD), gibberellic acid, salicylic acid, jasmonic acid, indoleacetic acid and abscisic acid were purchased from Sigma-Aldrich (MO, USA). Free fatty acids including C6:0, C8:0, C10:0, C12:0, C14:0, C16:0, C18:0, C20:0 and C22:0 were purchased from NU-CHEK PREP, Inc (MN, USA). All nanoparticles have been thermally treated at 350°C for 2 hours in a muffle furnace (Hubei, China) before use in order to remove trace organic contaminants.
5
Jasmonic Acid and Auxin Regulation of DAO Genes
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To check the effect of JA and IAA on the expression of DAO1 and DAO2, total RNA was extracted from whole seedlings of wild type and mutants (coi1-16 or jin1-2), which were grown under long-day conditions. Five days after germination, the seedlings were moved to sterile liquid media for overnight acclimation before any treatment. Jasmonic acid (Sigma-Aldrich, J2500, St. louis, MO, USA and IAA (Duchefa Biochemie, I0901) were used in this study. The auxinole was a gift from Professor Hayashi [33 (link)].
To check the expression of AOC2, OPR3, GH3.3, GH3.5, and GH3.6 genes, total RNA was extracted from etiolated hypocotyls. Wild-type and dao1-1 seedlings were first etiolated in the dark until their hypocotyls were 6 mm long (T0), and then they were transferred to long-day conditions for either 9 h (T9), 24 h (T24), or 72 h (T72).
To check the expression of AOC2, OPR3, GH3.3, GH3.5, and GH3.6 genes, total RNA was extracted from etiolated hypocotyls. Wild-type and dao1-1 seedlings were first etiolated in the dark until their hypocotyls were 6 mm long (T0), and then they were transferred to long-day conditions for either 9 h (T9), 24 h (T24), or 72 h (T72).
6
Jasmonic and Salicylic Acid Induction in Cotton
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Jasmonic acid (JA; Sigma, USA) was dissolved in a small quantity of ethanol, and further diluted with ddH2O to a concentration of 200 µmol·L−1. Salicylic acid (SA; Sigma, USA) was directly dissolved into heated ddH2O water to a final concentration of 400 µmol·L−1. Upland cotton cultivar CCRI 36 was planted in the greenhouse with an optimal temperature of 28°C. One-month-old plants were sprayed with JA and SA solutions when four true leaves had developed [20] , [21] (link), [36] (link). The top two leaves were removed at 0, 0.5, 6, 12, 24, and 48 h time-point. Harvested leaves were immediately plunged into liquid nitrogen, and stored at –80°C for later RNA extraction and relative transcript analysis. Nonexpressor of Pathogenesis-Related Genes 1 (GhNPR1, DQ409173), a SA- and JA-inducible gene in upland cotton was used as a positive control gene here [21] (link).
7
Preparation of Phytohormone Standards
8
Metabolomic Analysis of Plant Hormones
9
Phytohormone Modulation and Mealybug Interactions
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Chemical treatment: Jasmonic acid or salicylic acid (Sigma-Aldrich) was dissolved in 0.5 mL of acetone and dispersed in water (containing 0.1% Tween 20) to produce a 1.0 mM JA or SA solution. Each plant was sprayed with 1.0 mL/leaf of the JA or SA solution with a hand-sprayer. Twenty-four hours later, JA- or SA-treated plants were used for experiments.
Mealybug treatment: A mixture of third-instar nymphs and newly emerged adults (40 in total) of P. solenopsis were carefully transferred onto each plant and allowed to feed freely for 6, 12, 24, 72, or 120 h. After that, leaf samples were collected for phytohormone analysis. Plants pre-infested with 40 P. solenopsis adults (2- to 4-days old) for 120 h were used for performance bioassays, the EPG experiment, and gene-expression analysis.
Control treatment: In a preliminary experiment, we found that treatment with a water solution (including 0.5 mL of acetone and 0.1% Tween 20) had no effect on the performance of P. solenopsis reared on tomato plants. Thus, healthy and intact plants that received no treatment were used as control plants.
10
Plant Growth Regulator Preparation
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Karrikins (KAR1, KAR2), GR245DS, and GR24ent-5DS were prepared as described (Flematti et al., 2007 (link); Goddard-Borger et al., 2007 (link); Scaffidi et al., 2014 (link)) and dissolved as 10 mM stock solutions in acetone. Epibrassinolide (Sigma E1641), gibberellic acid (GA4 from L. N. Mander, Australian National University), 3-indoleacetic acid (Sigma I2886), (+)-cis, trans-abscisic acid (AG Scientific A-1103) and (±)-jasmonic acid (Sigma J2500) were dissolved in acetone as 5, 10, 10, 10, and 50 mM stock solutions, respectively.
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