Journal
FRONTIERS IN PLANT SCIENCE
Volume 13, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fpls.2021.715809
Keywords
wood; transcription factors; Y1H; Y2H; lignin; Populus
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Funding
- National Key Scientific Research Project of China [2021YFD2200205]
- National Natural Science Foundation of China [31670606, 31972955, 31770315, 32101549, 31972860]
- Taishan Scholar Program of Shandong
- First Class Grassland Science Discipline Program of Shandong Province
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Wood formation in trees is a complex process that involves protein-protein and protein-DNA interactions. In this study, we developed a high-throughput screening system to detect these interactions in wood development. Multiple regulatory modules of lignin biosynthesis were identified, and some interactions were further validated.
Wood formation of trees is a complex and costly developmental process, whose regulatory network is involved in the protein-protein and protein-DNA interactions. To detect such interactions in wood development, we developed a high-throughput screening system with 517 Gal4-AD-wood-associated transcription factors (TFs) library from Populus alba x P. glandulosa cv 84K. This system can be used for screening the upstream regulators and interacting proteins of targets by mating-based yeast-one hybrid (Y1H) and yeast-two-hybrid (Y2H) method, respectively. Multiple regulatory modules of lignin biosynthesis were identified based on this Populus system. Five TFs interacted with the 500-bp promoter fragment of PHENYLALANINE AMMONIA-LYASE 2 (PAL2), the first rate-limiting enzyme gene in the lignin biosynthesis pathway, and 10 TFs interacted with PaMYB4/LTF1, a key regulator of lignin biosynthesis. Some of these interactions were further validated by EMSA and BiFC assays. The TF-PaPAL2 promoter interaction and TF-PaMYB4 interaction revealed a complex mechanism governing the regulation of lignin synthesis in wood cells. Our high-throughput Y1H/Y2H screening system may be an efficient tool for studying regulatory network of wood formation in tree species.
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