Trunk twisting, a special phenomenon observed during plant growth, leads to remarkable changes in plant phenotypic plasticity and adaptability. However, it is a debate whether the trunk twisting belongs to environmental alteration or genetic variation. To understand the mechanisms underlying trunk twisting in Yunnan pine (Pinus yunnanensis Franch.), through a diallel cross experiment, we first determined that trunk twisting was controlled by recessive genes. Anatomical analysis identified that straight and twisty types differed significantly in xylem and phloem. RNA-seq of materials enriched by laser microdissection revealed several metabolic pathways with significant enrichment in twisty pines, including auxin signal transduction, photosynthetic carbon fixation and sucrose metabolism, etc. Application of exogenous auxin and auxin transport inhibitors (TIBA) can only change the growth rate, but cannot change the twisted or straight of the trunk. When auxin signaling inhibitors (auxinole) were added, the straight pines produce a tendency to twisty growth. Combined with enzyme activity assay and immunohistochemistry, we propose a working model. ARF can not only downregulate POR to block chlorophyll synthesis and photosynthesis but also upregulated Susy expression, and allows a large amount of sucrose to synthesize cellulose. Nevertheless, due to downregulated CBH expression and abnormal cellulolysis, cellulose accumulates and the lignin content decreases, eventually making the trunk highly prone to twisted growth. These results reveal the molecular mechanism of trunk twisting, and suggest that ARF expression method can be vital in trunk shape screening during the early growth stages of Yunnan pines.
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