The relict woody plant genus Liriodendron contains two endangered species, Liriodendron chinense and Liriodendron tulipifera. Understanding the molecular mechanisms involved in early embryo development is important for horticultural and ecological research, particularly for the development of improved somatic embryogenesis systems. However, the specific molecular processes underlying embryogenesis in these species remain largely unexplored. To address this, we investigated expression of the WOX (WUSCHEL-related homeobox) gene family of transcription factors throughout somatic embryogenesis. We confirmed expression of eight out of 11 novel candidate LcWOX genes in L.chinense using qRT-PCR and examined spatiotemporal expression patterns of the expressed genes using stable reporter lines that had been transformed with different LcWOX promoters driving GUS expression. We observed embryo developmental stages and expression patterns that broadly correlated with those reported for Arabidopsis somatic embryogenesis. LcWUS was weakly expressed during the transition stage and was predominantly restricted to the apical meristem. LcWOX5 was specifically expressed in the root meristem and restricted to the cotyledons thereafter, and LcWOX4 expression was restricted to the vascular tissue of cotyledonary embryos. In contrast, LcWOX9 was expressed in the embryonic callus and the entire embryonic cell mass, then became restricted to the basal cells, indicating a potential role in regulating embryonic maintenance. Our findings provide insights into spatiotemporally specific WOX transcription and shed new light on potential functions of WOX genes during Liriodendron somatic embryogenesis.

Roots are essential for plant growth, and studies on root-related genes, exemplified by WUSCHEL-RELATED HOMEOBOX5 (WOX5), have mainly concentrated on model organisms with less emphasis on the function of these genes in woody plants. Here, we report that overexpression of the WOX5 gene from Liriodendron hybrid (LhWOX5) in Arabidopsis leads to significant morphological changes in both the aerial and subterranean organs. In the Arabidopsis aerial parts, overexpression of LhWOX5 results in the production of ectopic floral meristems and leaves, possibly via the ectopic activation of CLV3 and LFY. In addition, in the Arabidopsis root, overexpression of LhWOX5 alters root apical meristem morphology, leading to a curled and shortened primary root. Importantly, these abnormal phenotypes in the aerial and subterranean organs caused by constitutive ectopic expression of LhWOX5 mimic the observed phenotypes when overexpressing AtWUS and AtWOX5 in Arabidopsis, respectively. Taken together, we propose that the LhWOX5 gene, originating from the Magnoliaceae plant Liriodendron, is a functional homolog of the AtWUS gene from Arabidopsis, while showing the highest degree of sequence similarity with its ortholog, AtWOX5. Our study provides insight into the potential role of LhWOX5 in the development of both the shoot and root.

The Pentatricopeptide repeat (PPR) superfamily is a large gene family in plants that regulates organelle RNA metabolism, which is important for plant growth and development. However, a genome-wide analysis of the PPR gene family and its response to abiotic stress has not been reported for the relict woody plant Liriodendron chinense. In this paper, we identified 650 PPR genes from the L. chinense genome. A phylogenetic analysis showed that the LcPPR genes could roughly be divided into the P and PLS subfamilies. We found that 598 LcPPR genes were widely distributed across 19 chromosomes. An intraspecies synteny analysis indicated that duplicated genes from segmental duplication contributed to the expansion of the LcPPR gene family in the L. chinense genome. In addition, we verified the relative expression of Lchi03277, Lchi06624, Lchi18566, and Lchi23489 in the roots, stems, and leaves and found that all four genes had the highest expression in the leaves. By simulating a drought treatment and quantitative reverse transcription PCR (qRT-PCR) analysis, we confirmed the drought-responsive transcriptional changes in four LcPPR genes, two of which responded to drought stress independent of endogenous ABA biosynthesis. Thus, our study provides a comprehensive analysis of the L. chinense PPR gene family. It contributes to research into their roles in this valuable tree species’ growth, development, and stress resistance.