Agarwood is a resinous wood of great economic value produced by trees from the Thymelaeaceae family in response to stress. The natural formation of agarwood can take decades after exposure to the stressors. Artificial agarwood induction by inoculating the stem with fungi has been successfully demonstrated, but resin accumulation occurs very slowly. Cell suspension and callus cultures may serve as an alternative solution to provide a fast-growing plant material to produce artificial agarwood in a short period. Here, we induced agarwood formation in callus cultures of Aquilaria malaccensis by application of crude mycelial extracts of Fusarium solani strains GSL1 or GSL2, or methyl jasmonate (MeJA). After 20 days of treatment with elicitors, all treated calluses had less dry weight than the control group. The gas chromatography–mass spectrometry analysis identified 33 different secondary metabolites among all samples, four of which were present in all treatments and control, i.e., 1-docosene and 1-octadecene (alkenes), 4-di-tert-buthylphenol (phenolic), and benzenepropanoic acid (fatty acid). The 6-methoxy-2-(4-methoxyphenethyl)-4H-chromene-4-one, a chromone derivative, was only detected in callus elicited with the F. solani strain GSL2 and MeJA. All treated calli produced more fatty acid derivatives than the control group. We conclude that elicitors used in this study can induce the production of agarwood-related chemicals such as chromone and fatty acid in callus culture.

Grapevine (Vitis vinifera L.) buds undergo seasonal dormancy to survive unfavourable conditions and synchronize growth with environmental cues. While dormancy transitions have been widely studied in temperate woody perennials, the physiological and metabolic dynamics underlying these transitions in grapevine remain poorly understood. Our study investigates seasonal changes in the physiology and metabolism of perennating buds in V. vinifera cv. Cabernet Sauvignon, focusing on dormancy depth, respiration, sugar metabolism, and cell cycle activity. We identified three distinct phases of bud quiescence: (i) para-dormancy (early summer), characterized by active metabolism and high levels of tricarboxylic acid cycle intermediates, shikimate, and myo-inositol; (ii) endo-dormancy (late summer to autumn), where dormancy depth peaked in late summer and was marked by reduced respiration and bud water content, and a sharp decline in hexose levels with a concomitant increase in raffinose; and (iii) eco-dormancy (winter to spring), featuring increased respiration, sugar mobilization (notably sucrose, glucose, and trehalose), and reactivation of cell division, with a shift of cells into the G2 phase. Just prior to bud burst, we observed a significant accumulation of sugar-phosphates, providing evidence that supports their role in promoting bud burst also in grapevine. Our findings provide new insights into the biochemical and physiological regulation of bud dormancy and bud burst, contributing to a deeper understanding of dormancy transitions in perennial crops.

The regulation of DNA accessibility by histone modification has emerged as a paradigm of developmental and environmental programming. Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a versatile tool to investigate in vivo protein–DNA interaction and has enabled advances in mechanistic understanding of physiologies. The technique has been successfully demonstrated in several plant species and tissues; however, it has remained challenging in woody tissues, in particular complex structures such as perennating buds. Here we developed a ChIP method specifically for mature dormant buds of grapevine (Vitis vinifera cv. Cabernet Sauvignon). Each step of the protocol was systematically optimized, including crosslinking, chromatin extraction, sonication and antibody validation. Analysis of histone H3-enriched DNA was performed to evaluate the success of the protocol and identify occupancy of histone H3 along grapevine bud chromatin. To our best knowledge, this is the first ChIP experiment protocol optimized for the grapevine bud system.