Non-Additive Gene Expression in Carbon and Nitrogen Metabolism Drives Growth Heterosis in Populus deltoides

Growth heterosis is crucial for Populus deltoides breeding, a key industrial-timber and ecological-construction tree species in temperate regions. However, the molecular mechanisms underlying carbon (C)-nitrogen (N) metabolism coordination in regulating growth heterosis remain unclear. Herein high-hybrids of P. deltoides exhibited high-parent heterosis and mid-parent heterosis in growth traits and key enzymes of C-N metabolism. In hybrids, gene expression patterns were mainly biased toward female parent. Parental contribution to growth heterosis in P. deltoides is differentiation, rather than absolute maternal or paternal dominance contributions. Parental genes were predominantly and dynamically inherited in a non-additive manner, mainly with dominant expression patterns. A total of 44 non-additive genes associated with photosynthetic C fixation, starch and sucrose metabolism, sucrose transport, photorespiration, and nitrogen metabolism coregulated growth heterosis by coordinating C-N metabolism. Growth-regulating factors 4 interacted with DELLA genes to promote growth by enhancing this coordination. Additionally, five critical genes were identified. Briefly, the above genes in high-hybrids improved photosynthesis and N utilisation by regulating carbohydrate accumulation and enzyme activity, while reducing respiratory energy consumption, thereby providing more energy for growth and promoting growth heterosis. Our findings offer new insights and theoretical basis for deep understanding genetic and molecular regulation mechanisms of tree heterosis and its application in precision hybrid breeding.