Phenols are bioactive substances of great interest because of their involvement in plant physiology, their use in many industrial processes, and their impact on human health. This work aims to summarize the varied approaches to the phenolic analysis of chestnut (bark and wood of trunk and branches, leaves, catkins, burs, and fruit) and to collate the optimal conditions into an easy to follow and execute protocol. Phenolic compounds were extracted by solid–liquid extraction and separated by liquid–liquid extraction. Total phenols content was determined by Folin–Ciocalteu assay, condensed tannins by vanillin assay, and hydrolyzable tannins (gallotannins and ellagitannins) by high-performance liquid chromatography quantification of methyl gallate and ellagic acid following acid methanolysis. The lowest temperature for conservation (–80 ℃), lyophilization, and milling (liquid N₂) were the most effective pretreatments for samples. For quantification of tannins, the use of water clearly reduced the sensitivity of the analysis of condensed tannins, whilst the more efficient degradation capacity of sulfuric acid improved the methanolysis of hydrolyzable tannins. These findings were validated using a range of chestnut tissues, and thus confirm the utility and effectiveness of this easy to implement, cost-effective, and efficient protocol.

Pinus pinaster is a plant species of great ecological and economic importance. Understanding the underlying molecular mechanisms that govern the growth and branching of P. pinaster is crucial for enhancing wood production and improving product quality. In this study, we describe a simple methodology that enables the discovery of candidate genes in Pinus pinaster by transferring existing knowledge from model species like Arabidopsis thaliana and focusing on factors involved in plant growth, including hormonal and non-hormonal pathways. Through comparative analysis, we investigated the main genes associated with these growth-related factors in A. thaliana. Subsequently, we identified putative homologous sequences in P. pinaster and assessed the conservation of their functional domains. In this manner, we can exclude sequences that, despite displaying high homology, lack functional domains. Finally, we took an initial approach to their validation by examining the expression levels of these genes in P. pinaster trees exhibiting contrasting growth patterns. This methodology allowed the identification of 26 candidate genes in P. pinaster. Our findings revealed differential expression patterns of key genes, such as NCED3, NRT1.2, PIN1, PP2A, ARF7, MAX1, MAX2, GID1, AHK4, AHP1, and STP1, in relation to the different growth patterns analyzed. This study provides a methodological foundation for further exploration of these genes involved in the growth and branching processes of P. pinaster. This will contribute to the understanding of this important tree species and open new avenues for enhancing its utilization in sustainable forestry practices.