Characterization of YABBY transcription factors in Osmanthus fragrans and functional analysis of OfYABBY12 in floral scent formation and leaf morphology.

BACKGROUND: The plant-specific YABBY transcription factor family plays important roles in plant growth and development, particularly leaf growth, floral organ formation, and secondary metabolite synthesis. RESULTS: Here, we identified a total of 13 OfYABBY genes from the Osmanthus fragrans genome. These 13 OfYABBY genes were divided into five subfamilies through phylogenetic analysis, and genes in the same subfamily showed similar gene structures and conserved protein motifs. Gene duplication promoted the expansion of the OfYABBY family in O. fragrans. Tissue-specific expression analysis showed that the OfYABBY family was mainly expressed in O. fragrans leaves and floral organs. To better understand the role of OfYABBY genes in plant growth and development, OfYABBY12 was selected for heterologous stable overexpression in tobacco, and OfYABBY12-overexpressing tobacco leaves released significantly fewer volatile organic compounds than wild-type tobacco leaves. Overexpression of OfYABBY12 led to the downregulation of NtCCD1/4 and decreased ß-ionone biosynthesis. Correspondingly, a dual-luciferase assay showed that OfYABBY12 negatively regulated the expression of OfCCD4, which promotes ß-ionone synthesis. Furthermore, tobacco leaves overexpressing OfYABBY12 were curled and wrinkled and had significantly reduced leaf thickness and leaf inclusions and significantly extended flower pistils (styles). CONCLUSION: Overall, the results suggest that the OfYABBY gene family may influence the biosynthesis of the floral scent (especially ß-ionone) in O. fragrans and may regulate leaf morphogenesis and lateral organs.

Floral Volatile Organic Compounds Change the Composition and Function of the Endophytic Fungal Community in the Flowers of Osmanthus fragrans.

Endophytic fungi in flowers influence plant health and reproduction. However, whether floral volatile organic compounds (VOCs) affect the composition and function of the endophytic fungal community remains unclear. Here, gas chromatography-mass spectrometry (GC-MS) and high-throughput sequencing were used to explore the relationship between floral VOCs and the endophytic fungal community during different flower development stages in Osmanthus fragrans 'Rixiang Gui'. The results showed that the composition of the endophytic fungal community and floral VOCs shifted along with flowering development. The highest and lowest α diversity of the endophytic fungal community occurred in the flower fading stage and full blooming stage, respectively. The dominant fungi, including Dothideomycetes (class), Pleosporales (order), and Neocladophialophora, Alternaria, and Setophoma (genera), were enriched in the flower fading stage and decreased in the full blooming stage, demonstrating the enrichment of the Pathotroph, Saprotroph, and Pathotroph-Saprotroph functions in the flower fading stage and their depletion in the full blooming stage. However, the total VOC and terpene contents were highest in the full blooming stage and lowest in the flower fading stage, which was opposite to the α diversity of the endophytic fungal community and the dominant fungi during flowering development. Linalool, dihydro-ß-ionone, and trans-linalool oxide(furan) were key factors affecting the endophytic fungal community composition. Furthermore, dihydro-ß-ionone played an extremely important role in inhibiting endophytic fungi in the full blooming stage. Based on the above results, it is believed that VOCs, especially terpenes, changed the endophytic fungal community composition in the flowers of O. fragrans 'Rixiang Gui'. These findings improve the understanding of the interaction between endophytic fungi and VOCs in flowers and provide new insight into the mechanism of flower development.

Structural characterization and anti-inflammatory activity of a pectin polysaccharide AP2-c from the lignified okra.

In this paper, a pectin polysaccharide AP2-c with molecular weight 6.69 × 105 Da was obtained from the lignified okra. The monosaccharide composition analysis indicated that AP2-c consisted of galactose, rhamnose and galacturonic acid in a molar ratio of 2.3: 1.5: 1.5. The structural characterization indicated that the main chain of AP2-c was composed of →2)-α-L-Rhap-(1→ and →4)-α-D-GalAp-(1→. →2)-α-L-Rhap-(1→ was branched at position O-4 and the branched chain consisted of →3,6)-ß-D-Galp-(1→, →6)-ß-D-Galp-(1→, α-L-Rhap-(1→ and ß-D-Galp-(1→. AP2-c could inhibit the mRNA expression levels of TNF-α, IL-1ß and iNOS in LPS-induced macrophages with a dose-dependent manner. Furthermore, AP2-c inhibited the phosphorylation of IκB and p65 via NF-κB pathway. The results indicated that AP2-c had obvious anti-inflammatory activity. PRACTICAL APPLICATIONS: When okra seeds were harvested, lignified okra was always abandoned as waste and had not been fully used for exploitation. Nevertheless, it accounted for more than half of the total plant's weight and was abundant in cell wall polysaccharides, which were the main components of okra to perform a variety of biological functions. In the research, the purified pectin polysaccharide AP2-c was obtained from lignified okra and its physicochemical properties, structural features and anti-inflammatory activity were systematically researched. It was detected that AP2-c exhibited anti-inflammatory activity by blocking NF-κB pathway and thus lowering the expression of related inflammatory factors. The results have significant implications for the value-added application of okra and its processing side products can obviously help to promote the anti-inflammatory application of AP2-c and avoid wasting resources.

Structural characterization and anti-inflammatory activity of a polysaccharide from the lignified okra.

The polysaccharide (AP1-b) of molecular weight 6.59 × 105 Da was isolated from lignified okra (Abelmoschus esculentus (L.) Moench) by hot-water extraction, 40 % ethanol precipitation and purified by DEAE Cellulose chromatography, respectively. The structure and anti-inflammatory activity of AP1-b were investigated. AP1-b was composed of galactose, rhamnose, gluctose, arabinose and galacturonic acid in a molar ratio of 1.98:1.00:0.15:0.32:0.29. The structural features showed that the AP1-b consisted of →2)-α-d-Rhap-(1→, →4)-ß-d-Galp-(1→, →4)-α-d-GalpA-(1→, →6)-ß-d-Galp-(1→, ß-d-Glcp-(1→ and α-l-Araf-(1→. AP1-b could observably improve the inflammatory injury of LPS-induced RAW 264.7 cells by inhibiting the secretion of NO and decreasing the levels of pro-inflammatory factors (IL-1ß, iNOS and TNF-α). AP1-b also inhibited the phosphorylation levels of IκB and p65 proteins, manifesting the anti-inflammatory activity of AP1-b may associated with inhibition of NF-κB signaling pathway. Therefore, AP1-b had potential value in treating inflammatory injury.

Purification, structural characterization and immunostimulatory activity of polysaccharides from Umbilicaria esculenta.

In this study, an active component UP1-1 was isolated from Chinese Huangshan Umbilicaria esculenta via hot water extraction and purified by anion-exchange and gel-filtration chromatography. UP1-1 mainly composed of galactose, mannose and glucose in a molar ratio of 0.8:1.0:4.6 with an average molecular weight of 281 kDa. Methylation analysis of UP1-1 revealed the major glycosidic bonds comprised 1,6-linked Glcp, 1,4-linked Glcp, t-linked Glcp, 1,3,6-linked Manp, 1,3-linked Galp, t-linked Galp at the ratio of 2.28:0.38:0.32:0.63:0.25:0.29. Structural analysis results revealed that the backbone of UP1-1 consisted of →6)-ß-D-Glcp-(1→, →6)-ß-D-Manp-(1→, →4)-ß-D-Glcp-(1 → residues with side chains of →3)-ß-D-Galp-(1→, ß-D-Galp-(1 → and ß-D-Glcp-(1 → branches located at O-3 position of →6)-ß-D-Manp-(1→. Immunostimulatory activity tests showed that UP1-1 could promote the phagocytic activity and NO production of RAW 264.7 cells in a dose-dependent manner. UP1-1 could significantly improve the proliferation effect of RAW 264.7 cells at the concentration of 50 µg/mL. Thus, UP1-1 exerted good immunostimulatory activity, suggesting that UP1-1 has a great potential application in pharmacological industry.