The ARF-Thiol Axis: An Epigenetic Model for the Biosynthesis and Domestication-Led Silencing of 3-Methyl-2-Butene-1-Thiol (3MB1T) in Cannabis sativa L.

"LEGAL NOTICE: This document constitutes an official Defensive Publication. The ARF-Thiol Axis and the Stress Hammer Protocol described herein are hereby placed in the public domain as Prior Art to prevent future patenting of these specific abiotic restoration mechanisms."This research dossier proposes the ARF-Thiol Axis, a novel mechanistic framework to account for the transgenerational phenotypic loss of high-potency volatile sulfur compounds (VSCs)—specifically 3-methyl-2-butene-1-thiol (3MB1T)—in Cannabis sativa L. We posit that the "Roadkill Skunk" (RKS) phenotype is a functional metabolic mutation characterized by the constitutive upregulation of specific Auxin Response Factors (CsARF21/22). This hormonal "overdrive" provides the transcriptional activation required for the GST-mediated conjugation and subsequent volatilization of thiols via the beta-lyase enzyme. Furthermore, we argue that decades of stable indoor domestication have triggered an "Epigenetic Lock," where DNA methylation silences these pathways in the absence of abiotic stressors. The dossier outlines a repeatable Abiotic-Trauma Protocol ("Stress Hammer") using targeted UV-B radiation and diurnal thermal shock to strip methylation marks and restore the ancestral phenotype. This framework is presented as a hypothesis-generating model for the global breeding and genomic research community to validate through GC-MS and bisulfite sequencing. Version 21.1 includes a strengthened Section X that broadens the prior-art framework to stress-responsive pathways across domesticated crops.