Background: Hosta plantaginea (Lam.) Aschers (HPA), a species in the family Liliaceae, is an important landscaping plant and herbaceous ornamental flower. However, because the flower has only two colors, white and purple, color matching applications are extremely limited. To date, the mechanism underlying flower color regulation remains unclear. Methods: In this study, the transcriptomes of three cultivars—H. plantaginea (HP, white flower), H. Cathayana (HC, purple flower), and H. plantaginea ‘Summer Fragrance’ (HS, purple flower)—at three flowering stages (bud stage, initial stage, and late flowering stage) were sequenced with the Illumina HiSeq 2000 (San Diego, CA, USA). The RNA-Seq results were validated by qRT-PCR of eight differentially expressed genes (DEGs). Then, we further analyzed the relationship between anthocyanidin synthase (ANS), chalcone synthase (CHS), and P450 and the flower color regulation by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Eukaryotic Orthologous Groups (KOG) network and pathway enrichment analyses. The overexpression of CHS and ANS in transgenic tobacco petals was verified using qRT-PCR, and the petal colors associated with the overexpression lines were confirmed using absorbance values. Results: Over 434,349 transcripts were isolated, and 302,832 unigenes were identified. Additionally, through transcriptome comparisons, 2098, 722, and 606 DEGs between the different stages were found for HP, HC, and HS, respectively. Furthermore, GO and KEGG pathway analyses showed that 84 color-related DEGs were enriched in 22 pathways. In particular, the flavonoid biosynthetic pathway, regulated by CHS, ANS, and the cytochrome P450-type monooxygenase gene, was upregulated in both purple flower varieties in the late flowering stage. In contrast, this gene was hardly expressed in the white flower variety, which was verified in the CHS and ANS overexpression transgenic tobacco petals. Conclusions: The results suggest that CHS, ANS, and the cytochrome P450s-regulated flavonoid biosynthetic pathway might play key roles in the regulation of flower color in HPA. These insights into the mechanism of flower color regulation could be used to guide artificial breeding of polychrome varieties of ornamental flowers.

Anthocyanins are natural colorants are synthesized in a branch of the flavonoid pathway. Dihydroflavonol-4reductase (DFR) catalyzes dihydroflavonoids into anthocyanins biosynthesis, which is a key regulatory enzyme of anthocyanin biosynthesis in plants. Hosta ventricosa is an ornamental plant with elegant flowers and rich colorful leaves. How the function of HvDFR contributes to the anthocyanins biosynthesis is still unknown. In this study, the DFR homolog was identified from H. ventricosa and sequence analysis showed that HvDFR possessed the conserved NADPH binding and catalytic domains. A phylogenetic analysis showed that HvDFR was close to the clade formed with MaDFR and HoDFR in Asparagaceae. Gene expression analysis revealed that HvDFR was constitutive expressed in all tissues and expressed highly in flower as well as was positively correlated with anthocyanin content. In addition, the subcellular location of HvDFR showed that is in the nucleus and cell membrane. Overexpression of HvDFR in transgenic tobacco lines enhanced the anthocyanins accumulation along with the key genes upregulated, such as F3H, F3ʹH, ANS, and UFGT. Our results indicated a functional activity of the HvDFR, which provide an insight into the regulation of anthocyanins content in H. ventricosa.

The Hosta hybrid cultivar ‘So Sweet’, an important ornamental and widely used horticultural plant, is noted for its rich, fragrant white flowers. The main aroma components of Hosta flowers are terpenoids, mainly monoterpenes. Until now, the terpene synthases responsible for terpene production in Hosta were not described. In this study, two terpene synthase (TPS) genes (HsTPS1 and HsTPS2) were cloned and characterized to further study their function. Furthermore, the volatile terpenes of Hosta ’So Sweet’ in two flower development stages from two in vitro enzyme tests were analyzed by gas chromatography–mass spectrometry (GC–MS). We analyzed the expression levels of two genes at four different developmental stages using quantitative real-time PCR, while localization was analyzed using Nicotina benthamiana leaves. In vitro, the two proteins were identified to mainly produce linalool and nerol. In addition, the active products of the two recombinant proteins were (E,E)-farnesol and (E,E)-farnesal, respectively, using farnesyl pyrophosphate as a substrate. The high expression of HsTPS1 and HsTPS2 was correlated with the release of components of Hosta flowers. To our knowledge, this is the first time that the terpene synthase genes of Hosta species have been isolated and identified, providing an opportunity to study the terpene metabolic pathways in Hosta species.

Seabuckthorn (Hippophae rhamnoides Linn.) is a commonly utilized medicinal crop with various applications in the treatment of different diseases. Two particularly noteworthy nutrients in seabuckthorn fruit are seabuckthorn oil and flavonoids. In recent years, seabuckthorn oil has attracted considerable attention due to its perceived benefits for beauty and healthcare. Consequently, there is a clear need for further research into seabuckthorn oil. While numerous studies have been conducted on the regulation of oil by the FAD2 gene family, there is a paucity of literature examining the molecular mechanism of FAD2 gene involvement in seabuckthorn oil regulation. Accordingly, two FAD2 genes have been identified in seabuckthorn, which are classified differently and perform distinct functions. Both genes are located in the endoplasmic reticulum. Following transient expression in seabuckthorn fruits, it was observed that HrFAD2-1 and HrFAD2-3 were capable of influencing the synthesis of α-linolenic acid, with HrFAD2-1 additionally demonstrated to facilitate the synthesis of lysophosphatidic acid. All of the aforementioned genes have been observed to promote jasmonic acid (JA) synthesis. The heterologous transformation of Linum usitatissimum demonstrates that both HrFAD2-1 and HrFAD2-3 are capable of promoting plant growth. The HrFAD2-1 gene was observed to significantly increase the content of major fatty acids in Linum usitatissimum Linn seeds, whereas the HrFAD2-3 gene appeared to be primarily involved in the regulation of plant growth and development. In conclusion, a preliminary investigation into the functions of the HrFAD2-1 and HrFAD2-3 genes in fatty acid synthesis was conducted. This revealed that HrFAD2-1 is closely associated with oleic acid synthesis and acts as a negative regulator. Furthermore, our findings will provide a foundation for subsequent investigations into the fatty acid synthesis pathway in Hippophae rhamnoides oil, offering a theoretical basis for subsequent studies at the molecular level.