Mapping the dynamic plant interactome: from in vitro assays to in vivo quantitative approaches
Rattachement africain : cn, tr. Niveau de preuve : code pays fourni par la source.
Le résumé fourni par la source
BACKGROUND: Protein-protein interactions underpin virtually all biological processes in plants, from signal transduction and immune responses to development and stress adaptation. Despite their fundamental importance, the plant interactome remains far from complete, and existing maps are systematically biased by the technical limitations inherent to conventional detection platforms. MAIN BODY: This review critically traces the evolution of protein-protein interaction methodologies, from foundational approaches to advance in vivo and quantitative platforms. Classical techniques such as the yeast two-hybrid system and in vitro pull-down assays operate outside physiological cellular environments and are poorly suited to capturing transient or condition-dependent interactions. Affinity purification coupled with mass spectrometry improves throughput but remains vulnerable to artifacts introduced during cell lysis and to the preferential loss of weak interactors. To address these shortcomings, proximity labeling with engineered biotin ligases, most notably the fast-acting variant TurboID, has emerged as a powerful strategy, enabling covalent biotinylation of protein neighborhoods within living cells prior to lysis and thereby preserving associations that conventional methods routinely miss. Because TurboID reports proximity rather than direct binding, its output requires downstream binary validation. Complementary in planta validation tools are equally critical for moving beyond discovery. Split-luciferase complementation assays based on the NanoLuciferase reporter provide exceptional sensitivity for binary interaction detection under native expression conditions, while Förster Resonance Energy Transfer measured through fluorescence lifetime imaging microscopy offers quantitative biophysical evidence of molecular proximity at endogenous expression levels, serving as a high-confidence validation approach. Emerging technologies, including high-throughput protein microarrays and optogenetically controlled dimerization systems, further expand the methodological repertoire available to the plant biology community. CONCLUSION: We propose a practical, integrative three-tier framework, combining proximity labeling for broad in vivo discovery, split-luciferase complementation for sensitive binary validation, and fluorescence lifetime imaging microscopy for quantitative confirmation, that systematically funnels candidate interactions from initial identification to physiologically rigorous verification. This framework synthesizes established best practices into a structured workflow applicable to mapping dynamic plant interactomes, though its optimal implementation will depend on the biological question, target protein class, and available resources.
Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.
Le contrôle bibliographique ouvert
DOI retrouvé dans Crossref DOI retrouvé ; titre concordant.
- Titre Crossref
- Mapping the dynamic plant interactome: from in vitro assays to in vivo quantitative approaches
- Date Crossref
- 12/07/2026
- Éditeur
- Springer Science and Business Media LLC
- Type
- journal-article
Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.
Où se fait cette recherche
-
Huazhong Agricultural University Hongshan Laboratory pays non établi dans la noticeUniversité ou école supérieure
-
Xinjiang Production and Construction Corps pays non établi dans la noticeOrganisation à but non lucratif
-
Tarim University Xinjiang Production and Construction Corps Key Laboratory of Facility Agriculture pays non établi dans la noticeUniversité ou école supérieure
-
Yangzhou University pays non établi dans la noticeUniversité ou école supérieure
-
Akdeniz University pays non établi dans la noticeUniversité ou école supérieure
-
College of Plant Science and Technology MOA Key Laboratory of Crop Ecophysiology and Farming System in the Middle Reaches of the Yangtze River pays non établi dans la noticeUniversité ou école supérieure
-
College of Horticulture and Forestry Science National Key Laboratory for Germplasm Innovation and Utilization of Horticultural Crops pays non établi dans la noticeUniversité ou école supérieure
-
College of Horticulture and Landscape Architecture pays non établi dans la noticeUniversité ou école supérieure
-
Faculty of Agriculture Department of Plant Breeding and Genetics pays non établi dans la noticeUniversité ou école supérieure
-
Agricultural Institute Department of Plant Physiology and Metabolomics pays non établi dans la noticeStructure de recherche
Hongshan Laboratory — Huazhong Agricultural University, Xinjiang Production and Construction Corps et Xinjiang Production and Construction Corps Key Laboratory of Facility Agriculture — Tarim University, avec 7 autres affiliations.
Une affiliation ne permet pas de déduire la nationalité d’un auteur.