Chilling and gibberellin acids hyperinduce β-1,3-glucanases to reopen transport corridor and break endodormancy in tree peony (Paeonia suffruticosa)

Bud endodormancy is accompanied by transport channel apertures blockage through callose deposition, and its resume to growth requires evoking β-1,3-glucanases (BGs) to unchoke the conduit. To understand out its working manner, the statuses of the transport channels were evaluated and candidate BGs were identified during chilling and gibberellin acids (GA) induced dormancy release in tree peony. Calcein reflects plasmodesmata permeability, and no calcein was observed in the bud together with density aniline blue fluorescent around the stem phloem at 0 d chilling. With the increase of chilling accumulation, the contents of glucan declined and the activities of gulcanase increased gradually in buds, and the calcein reached the top of flower primordia at 21 d chilled bud. Both GA3 and GA4 feedings promoted bud sprouting and growth along with rapidly unchoking the transport channels, and GA3 was more effective. Several candidate β-1,3-glucanase genes were detected, combining transcriptional profiling and quantitative PCR analysis. PsBG1, PsBG3, PsBG6, PsBG8 and PsBG9 were inducible by chilling accumulation and presented laminarin hydrolyzing activities after prokaryotically expression, while PsBG1, PsBG3, PsBG8 and PsBG9 responded to GAs application. Subcellular localizations revealed that PsBG6 and PsBG9 were plasmodesmata residents. It was concluded that PsBG6 played a vital role in chilling accumulation response and PsBG9 was central in GAs-induced dormancy release, and they could be used as marker genes for dormancy release in tree peony. These results were of great value to understand the mechanism of dormancy regulation and as an important fundamental for forcing culture technology in tree peony.