Carotenoid cleavage oxygenase (CCO) is an enzyme that can catalyze carotenoids to volatile aromatic substances and participate in the biosynthesis of two important phytohormones, i.e., abscisic acid (ABA) and strigolactone (SL). However, the genome-wide identification and analysis of the CCO gene family in the rare and endangered woody plant Liriodendron chinense has not been reported. Here, we performed a genome-wide analysis of the CCO gene family in the L. chinense genome and examined its expression pattern during different developmental processes and in response to various abiotic stresses. A total of 10 LcCCO genes were identified and divided into 6 subfamilies according to the phylogenetic analysis. Subcellular localization prediction showed that most of the LcCCO proteins were located in the cytoplasm. Gene replication analysis showed that segmental and tandem duplication contributed to the expansion of this gene family in the L. chinense genome. Cis-element prediction showed that cis-elements related to plant hormones, stress and light response were widely distributed in the promoter regions of LcCCO genes. Gene expression profile analysis showed that LcNCED3b was extensively involved in somatic embryogenesis, especially the somatic embryo maturation, as well as in response to heat and cold stress in leaves. Furthermore, qRT-PCR analysis showed that LcNCED3b obviously responded to drought stress in roots and leaves. This study provides a comprehensive overview of the LcCCO gene family and a potential gene target for the optimization of the somatic embryogenesis system and resistance breeding in the valuable forest tree L. chinense.

Chloroplasts is the site for photosynthesis, which is the main primary source of energy for plants. Golden2-like (GLK) is a key transcription factor that regulates chloroplast development and chlorophyll synthesis. However, most studies on GLK genes are performed in crops and model plants with less attention to woody plants. In this study, we identified the LhGLK1 and LhGLK2 genes in the woody plant Liriodendron hybrid, and they are specifically expressed in green tissues. We showed that overexpression of the LhGLK1 gene improves rosette leaf chlorophyll content and induces ectopic chlorophyll biogenesis in primary root and petal vascular tissue in Arabidopsis. Although these exhibit a late-flowering phenotype, transgenic lines accumulate more biomass in vegetative growth with improved photochemical quenching (qP) and efficiency of photosystem II. Taken together, we verified a conserved and ancient mechanism for regulating chloroplast biogenesis in Liriodendron hybrid and evaluated its effect on photosynthesis and rosette biomass accumulation in the model plant Arabidopsis.

In this study, 52 AAAP genes were identified in the L. chinense genome and divided into eight subgroups based on phylogenetic relationships, gene structure, and conserved motif. A total of 48 LcAAAP genes were located on the 14 chromosomes, and the remaining four genes were mapped in the contigs. Multispecies phylogenetic tree and codon usage bias analysis show that the LcAAAP gene family is closer to the AAAP of Amborella trichopoda, indicating that the LcAAAP gene family is relatively primitive in angiosperms. Gene duplication events revealed six pairs of segmental duplications and one pair of tandem duplications, in which many paralogous genes diverged in function before monocotyledonous and dicotyledonous plants differentiation and were strongly purification selected. Gene expression pattern analysis showed that the LcAAAP gene plays a certain role in the development of Liriodendron nectary and somatic embryogenesis. Low temperature, drought, and heat stresses may activate some WRKY/MYB transcription factors to positively regulate the expression of LcAAAP genes to achieve long-distance transport of amino acids in plants to resist the unfavorable external environment. In addition, the GAT and PorT subgroups could involve gamma-aminobutyric acid (GABA) transport under aluminum poisoning. These findings could lay a solid foundation for further study of the biological role of LcAAAP and improvement of the stress resistance of Liriodendron.

Auxin, the first discovered phytohormone, is important for the growth and development of plants through the establishment of homeostasis and asymmetry. Here, we cloned the auxin transporter gene PIN-FORMED3 (PIN3) from the valuable timber tree hybrid Liriodendron (Liriodendron chinense × Liriodendron tulipifera). The gene contained a complete open reading frame of 1917 bp that encoded 638 amino acids. Phylogenetic analysis indicated that LhPIN3 exhibited the highest sequence similarity to the PIN3 of Vitis vinifera. Quantitative real-time PCR analysis showed that LhPIN3 was broadly expressed across different tissues/organs of Liriodendron, with the highest expression level in the roots. Heterologous overexpression of LhPIN3 in Arabidopsis thaliana caused considerable phenotypic changes, such as the root length and number of flowers. Genetic complementation of Arabidopsis pin1 mutants by LhPIN3, driven by the cauliflower mosaic virus 35S promoter, fully restored the root length and number of flowers of the pin1 mutant. Overall, our findings reveal that LhPIN3 has similar capacities to regulate the root length and number of flowers of Arabidopsis with AtPIN1.

Liriodendron is a highly valued ornamental genus renowned for its distinctive tulip-shaped flowers. Despite its horticultural importance, the molecular mechanisms underlying interspecific variation in petal coloration, namely green petals in Liriodendron chinense (Hemsl.) Sargent, an orange-yellow basal band in Liriodendron tulipifera Linn., and an extended orange-yellow band in their hybrid, remain poorly understood. By integrating morphological, transcriptomic, and metabolomic analyses, we found that orange-yellow pigmentation during petal development is closely associated with chlorophyll degradation and carotenoid biosynthesis. The expression of chlorophyll synthesis genes PORA1 and PORA2 showed a strong positive correlation with chlorophyll content, and their downregulation led to disrupted chloroplast structure and reduced chlorophyll levels. Concurrently, carotenoid biosynthesis genes CRTISO and LCYE were markedly upregulated during the formation of the colored petal band. These results highlight the synergistic roles of chlorophyll and carotenoid metabolism in determining petal color patterning in Liriodendron, providing a genetic basis for the targeted breeding of ornamental traits.