Z 3 hexenol

The chemicals used for volatile identification and enzyme activity, including hexyl acetate, E-2-hexenyl acetate, Z-3- hexenyl acetate, hexanol, E-2-hexenol, Z-3-hexenol, 2-octanol, butyl acetate, geranyl acetate, ethyl hexanoate, ethyl benzoate, MeJA, MeSA, JA, and SA were purchased from Sigma-Aldrich (USA).

Two-week-old N. benthamiana plants grown on Murashige and Skoog agar plates (250 cm³) were treated with dichloromethane (DCM; Sigma–Aldrich) or individual GLVs (i.e., E-2-hexenal, Z-3-hexenol, and Z-3-hexenyl acetate; Sigma–Aldrich). Volatiles were diluted with DCM, which does not induce HPL expression. A 2-μL aliquot of 0.1 M volatile solution was applied to 3M^TM Micropore^TM Surgical Tape, which was attached to the inside of the plate cover. The cover was immediately set on the plastic plate, and the plants were incubated for 1 h at 25°C in an illuminated growth chamber (70 μmol m^-2 s^-1). DCM-treated plants were used as controls.

Female pheromone components were selected based on their ability to elicit single sensillum responses in female H. virescens moths [11 ]. The female sex pheromone components were as follows: (Z)-9-tetradecenal (Z9-14:Ald), (Z)-11-hexadecenal (Z11-16:Ald) and (Z)-11-hexadecenyl acetate (Z11-16:OAc) and were obtained from Bedoukian Research, Inc. (Danbury, Connecticut, USA). Putative host volatiles were selected from compounds that have shown physiological or behavioral effects on H. virescens in previous studies [11 ][12 ][39 (link)][40 (link)]: 2-phenyl ethanol, Z3-hexenol, along with racemic linalool and β-caryophyllene were obtained from Sigma Aldrich (St. Louis, MO, USA). All chemicals were diluted in hexane at decade steps from 10ng/μl-100μg/μl, and stored at -20°C. OA-hydrochloride was obtained from Sigma Aldrich (St. Louis, MO, USA) and stored at room temperature. OA was diluted to 50μg/μl in hemolymph Ringer solution [41 ] and stored in a fridge at 4°C.

Unless otherwise stated, all the reagents used in this study were analytically pure. The water used for various experiments was purified with a Milli-Q purification system. Tween 80, glucose anhydrous, cupric sulfate, potassium sodium tartrate tetrahydrate, sodium hydroxide, magnesium chloride, acetyl-CoA, linoleic acid sodium salt, nicotinamide adenine dinucleotide, dithiothreitol, Triton X-100, 4-morpholineethanesulfonic acid, polyvinylpolypyrrolidone (PVPP), dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium acetate, acetaldehyde, and butanol were obtained from Yuanye Co., Ltd. (Shanghai, China). BTH, E-2-hexenal, hexanal, Z-3-hexenal, E-2-hexenol, Z-3-hexenol, hexanol, Z-3-hexenyl acetate, hexyl acetate, 2-octanol, linolenic acid, and linoleic acid were HPLC grade and obtained from Sigma-Aldrich (Shanghai, China).

n-Alkanes (C8–C40) were purchased from J&K Scientific, Beijing, China. Benzaldehyde, benzyl alcohol, (Z)-3-hexenyl acetate, methyl salicylate, and 2-phenylethanol were purchased from Wako Pure Chemical Industries Ltd, Osaka, Japan. Ethyl n-decanoate, geraniol, 1-hexanol, (Z)-3-hexenol, phenylacetaldehyde, α-farnesene, (E)-nerolidol, linalool, linalool oxide, polyvinylpolypyrrolidone (PVPP), and XAD-2 were purchased from Sigma-Aldrich Company Ltd., Louis, MO, USA. geraniol was purchased from TCI Development Co., Ltd, Shanghai, China. The Quick RNA isolation kit was purchased from Huayueyang Biotechnology Co., Ltd., Beijing, China. The 2× SYBR Green Universal PCR Mastermix was purchased from Bio-Rad Laboratories, Richmond, CA, USA.

LiChrolut EN cartridges (500 mg, 6 mL) were obtained from Merck (Darmstadt, Germany). All chemicals were of analytical reagent grade unless otherwise stated, and water was obtained from a Milli-Q purification system (MilliporeSigma, Burlington, MA, USA). Folin-Ciocalteu reagent, sodium carbonate, sodium acetate, potassium chloride, sodium chloride, sucrose, methanol (HPLC grade), ethanol (HPLC grade), dichloromethane (HPLC grade) and gallic acid were purchased from SCR^® (Shanghai, China). Eucalyptol (99%), linalool (≥95%), (−)-myrtenol (95%), carveol (97%, mixture of isomers), borneol, (≥99.0%, sum of enantiomers, GC), terpinolene (≥94.0%), β-citronellol (95%), geraniol (98%), ethyl-2-methylbutyrate (99%), ethyl-3-methylbutyrate (98%), (Z)-3-hexenol (98%), (E)-2-hexenol (96%), benzyl alcohol (≥99%), phenylethyl alcohol (≥99%), cuminic alcohol (97%), methyl butanoate (99%), isobutyl acetate (99%), ethyl butanoate (99%), isoamyl acetate (≥99%), methyl benzoate (99%), ethyl benzoate (≥99%), diethyl succinate (99%), methyl salicylate (≥99%), ethyl octanoate (≥99%), ethyl decanoate (98%), methyl vanillate (99%), decanoic acid (≥98.0%), benzaldehyde (≥99%), p-cresol (99%), 4-vinylguaiacol (≥98.0%), vanillin (99%), (E)-asarone (98%), (Z)-asarone (70%) were obtained from Sigma-Aldrich (St. Louis, MO, USA). All volatile standards were prepared by dilution with HPLC grade methanol.