Abiotic stress treatment reveals expansin like A gene OfEXLA1 improving salt and drought tolerance of Osmanthus fragrans by responding to abscisic acid

Sweet osmanthus(Osmanthus fragrans) is a having general approval aromatic tree in China that is widely applied to landscaping and gardening. However, the evergreen tree adaptability is limited by many environmental stresses. Currently, limited information is available regarding the genetic analysis and functional identification of expansin genes in response to abiotic stress in sweet osmanthus. In this study, a total of 29 expansin genes were identified and divided into four groups by genome-wide analysis from the sweet osmanthus genome. Transcriptome and quantitative Real-time PCR analysis showed that the cell wall-localized protein expansin-like A(OfEXLA1) gene was significantly induced by salt and drought treatment. Histochemical GUS staining of transgenic Arabidopsis lines in which GUS activity was driven with the OfEXLA1 promoter, GUS activity was significantly induced by salt, drought, and exogenous abscisic acid(ABA). In yeast, we found OfEXLA1overexpression significantly improved the population of cells compared with wild-type strains after NaCl and polyethylene glycol(PEG)treatment. Additionally, OfEXLA1 overexpression not only promoted plant growth, but also improved the salt and drought tolerance in Arabidopsis. To gain insight into the role of ABA signaling in the regulation of OfEXLA1 improving abiotic tolerance in sweet osmanthus, four differentially expressed ABA Insensitive 5(ABI5)-like genes(OfABL4, OfABL5, OfABL7, and OfABL8) were identified from transcriptome, and dualluciferase(dual-LUC) and yeast one hybrid(Y1H) assay showed that OfABL4 and OfABL5 might bind to OfEXLA1 promoter to accumulate the OfEXLA1 expression by responding to ABA signaling to improve abiotic tolerance in sweet osmanthus. These results provide the information for understanding the molecular functions of expansin-like A gene and molecular breeding of sweet osmanthus in future.

Study on the molecular mechanism of Osmanthus fragrans Lour.on boosting immunity based on network pharmacology and molecular docking

Objective:To explore the molecular mechanism of Osmanthus Fragrans Lour.(OFL)in enhancing immunity.Methods:The compounds and action targets of OFL were collected from the Traditional Chinese Medicine Systematic Pharmacology Database and Analysis Platform.Protein targets of compounds were obtained from the UniProt database and relevant targets of boosting immunity were retrieved from the Genecards database.The Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis and Gene Ontology function enrichment analysis were performed through the DAVID analysis website,Visualization and Integrated Discovery.Finally,the results of the network analysis were validated by performing molecular docking using AutoDock vina.Results:A total of 7 active compounds and 167 potential active targets were identified in OFL.A total of 1549 genes with a correlation score of≥1 were retrieved from the Genecards website with the keyword“boost immunity”,and 107 genes were obtained by crossing the 167 genes of OFL with the 1549 genes of boosting immunity.A total of 4802 entries were obtained from Gene Ontology functional enrichment(P<0.05).A total of 234 signaling pathways were obtained through a Kyoto Encyclopedia of Genes and Genomes pathway analysis(P<0.05).Tumor necrosis factor(TNF)and interleukin 17(IL-17)signaling pathways were closely related to body immunity.The molecular docking results showed that all the core compounds in OFL the characteristics including low energy,a stable structure and high binding activity when bound to IL-17 and TNF-αprotein.Kaempferol showed the highest affinity with IL-17,and fucosterol showed the highest affinity with TNF-α.Conclusions:Through studies on network pharmacology and molecular docking,we have further demonstrated that OFL could enhance the immunity of the body through multi-component,multi-target and multi-pathway actions,and that IL-17/TNF-αsignalling pathway is the key molecular mechanism.

Construction of China Osmanthus City and Intellectual Property Right Protection in Xianning City in the Context of Three New Developments

Xianning City of Hubei Province is one of the five major traditional osmanthus producing areas in the world and the hometown of osmanthus in China. Xianning is constructing China Osmanthus City in the whole city. This paper first introduces the main characteristics of the osmanthus industry in Xianning City, and analyzes the characteristics of traditional knowledge, biological genetic resources, new plant varieties, patents, geographical indications, and trademarks. Then, it studies the intellectual property resources of the osmanthus industry, and analyzes the main problems existing in the construction of China Osmanthus City and intellectual property right protection. Finally, it comes up with constructive recommendations including applying for China and Globally Important Agricultural Heritage Systems, developing colored-leaf osmanthus varieties, creating a regional public brand of osmanthus specialty products, and building a new development pattern of osmanthus industry.

Analysis on Agricultural Meteorological Condition of Osmanthus Blooming

Osmanthus, an ornamental plants, is favored by people, and forecast of Osmanthus florescence is of vital significance for enjoying the sight. Based on 15 years of flowering observation data, analysis was made on agricultural meteorologi- cal conditions of Osmanthus in Guilin and the results showed that ground tempera- ture, humidity, precipitation, sunshine have significant effects on Osmanthus flores- cence. In forecasting florescence, therefore, it is necessary to take comprehensive consideration of weather conditions and changes in the future, to release Osman- thus flowering forecast as per method of combination of long-term forecasting and short-term forecast.

Characteristics of Uptake and Accumulation of Pb and Cd in Osmanthus fragrans(Thunb.) Lour.under Single and Combined Stresses of Pb and Cd

[Objective] This study aimed to investigate the characteristics of Pb and Cd uptake and accumulation in Osmanthus fragrans（Thunb.） Lour.under single and combined stress of Pb and Cd.[Method] Pot experiment was adopted with O.fragrans seedlings as experimental materials to investigate the effect of single and combined stresses of Pb and Cd on the growth of O.fragrans seedlings under 0,50,100,500,1 000 and 2 000 mg/kg of Pb and 0,1,10,50,100 and 200 mg/kg of Cd.[Result] Under the experimental conditions,the growth of O.fragrans seedlings was significantly inhibited under moderate Pb stress（500 mg/kg） and high Cd stress（100 mg/kg）;the decreasing order of Pb uptake rate of different O.fragrans tissues was root stem leaf;the decreasing order of Cd content of different O.fragrans tissues was root stem leaf,while the decreasing order of Cd uptake rate of different O.fragrans tissues was stem root leaf.[Conclusion] Under combined stress,low concentrations of Pb and Cd can mutually promote the uptake and accumulation,while high concentrations of Pb and Cd can mutually inhibit the uptake and accumulation.

Plant Landscaping Characteristics of the Sweet Osmanthus Exposition Park in Guilin City

Through field investigation, the plant landscaping and its characteristics of Sweet Osmanthus Exposition Park in Guilin were analyzed in terms of three aspects: the allocation of Sweet Osmanthus species, the collocation of Sweet Osmanthus and other plants, plants' arrangement in Sweet Osmanthus landscape garden, which would provide reference for the construction of the special categorized plants gardens.

Differences in leaf functional traits between red and green leaves of two evergreen shrubs Photinia×fraseri and Osmanthus fragrans

Leaf functional traits are adaptations that enable plants to live under different environmental conditions. This study aims to evaluate the differences in leaf functional traits between red and green leaves of two evergreen shrubs Photinia × fraseri and Osrnanthus fragrans. Specific areas of red leaves are higher than that of green leaves in both species. Thus, the material investment per unit area and per lamina of red leaves is significantly lower than that of green leaves, implying an utmost effort of red leaves to increase light capture and use efficiency because of their low leafchlorophyll concentration. The higher petiole length of green leaves compared with that of red leaves indicates that adult green leaves may have large fractional biomass allocation to support the lamina structures in capturing light with maximum efficiency and obtaining a high growth rate. The high range of the phenotypic plasticity of leaf size, leaf thickness, single-leaf wet and dry weights, and leaf moisture of green leaves may be beneficial in achieving efficient control of water loss and nutrient deprivation. The high range of phenotypic plasticity of leaf chlorophyll concentration of red leaves may be advantageous in increasing resource （especially light） capture anduse efficiency because this leaf type is juvenile in the growth stage and has low leaf-chlorophyll concentration.

Identification and quantification of the bioactive components in Osmanthus fragrans roots by HPLC-MS/MS

The roots of O. fragrans are also a valuable resource in addition to its flowers and fruits. In this study, the HPLC-MS/MS method used for analyzing the chemical constituents in O. fragrans roots extract was developed, which showed high sensitivity for both qualitative and quantitative analyses. Thirty-two compounds were first discovered in O. fragrans roots, one compound of which was reported for the first time. The simultaneous determination method for acteoside, isoacteoside, oleuropein and phillyrin was validated to be sensitive and accurate. Then it was applied to determine the content of bioactive components in O. fragrans roots from different cultivars. The content of oleuropein and phillyrin in the twelve batches was relatively stable, while the content of acteoside and isoacteoside varied greatly.Moreover, the therapeutic material basis and mechanism of O. fragrans roots exerting its traditional pharmacodynamics were analyzed by network pharmacology. The results showed that O. fragrans roots might be effective for the treatment of inflammation, cardiovascular diseases, cancer, and rheumatoid arthritis, which is consistent with the traditional pharmacodynamics of O. fragrans roots. This work can provide an analytical method for the comprehensive development of O. fragrans roots.

CPTA treatment reveals potential transcription factors associated with carotenoid metabolism in flowers of Osmanthus fragrans

Osmanthus fragrans is one of the top ten traditional flowers in China.It is divided into three different groups according to its color.α-Carotene and β-carotene are the main determinants to distinguish the color differences between three groups.However,the dominant genes and transcription factors involved in carotenoid metabolism remain unclear.CPTA treatment(0.7mmol·L−1)remarkably promoted lycopene,α-carotene and β-carotene contents in flowers.Transcriptome sequencing analysis revealed that CPTA treatment could trigger chain reactions in carotenoid metabolism pathway genes.Four up-regulated and 10 down-regulated transcription factors which have close association with carotenoid variation were significantly induced by CPTA treatment.The up-regulated TFs such as MYB43,MYB123,HSF,were further subjected to transcript expression determination in different cultivars with drastic colors.Among them,transcript expression of four up-regulated TFs coincided with the carotenoid accumulation in different cultivars.We selected up-regulated OfMYB43 to verify its function,which is related to stress tolerance and transcriptional regulation.Transient overexpression of OfMYB43 in O.fragrans flowers showed that it could remarkably promote the expression of PDS,ZISO,LCYE and CCD4,leading to increased accumulation of β-branch carotenoids.OfMYB43 was a potential positive regulator of carotenoid biosynthesis in O.fragrans flowers.This study provides insight into the molecular mechanism of carotenoid metabolism in O.fragrans.

First Report of Botryosphaeria dothidea Causing Sweet Osmanthus Leaf Dieback in China

Sweet osmanthus is one of the ten traditional famous flowers in China.The occurrence of the diseases caused by fungi other than Botryosphaeria spp.has been reported mainly from China on sweet osmanthus.A leaf dieback of sweet osmanthus caused by Botryosphaeria sp.was found for the first time in 2007 in Nanning City,Guangxi,China.The objectives of the present study were to isolate and characterize the causal organism of sweet osmanthus leaf dieback.The fungus was isolated from the lesions of affected sweet osmanthus leaves and its pathogenicity to sweet osmanthus was confirmed using a detached-leaf-inoculation method.The identification of the pathogen was carried out mainly based on the morphological characters and molecular analysis of internal transcribed spacer （ITS） sequences.The morphological characters of the pathogenic isolate GHX6 were agreed with that of Botryosphaeria dothidea.The ITS sequence of the isolate was amplified with primers ITS1 and ITS4,and submitted to GenBank （accession no.GQ368251）.Molecular analysis based on ITS sequence comparison between the isolate GHX6 and the other related fungi derived from GenBank supported that the causal agent of the sweet osmanthus leaf dieback belonged to Botryosphaeria dothidea.This is the first report of Botryosphaeria dothidea causing leaf dieback on sweet osmanthus in China.

cDNA Cloning and Expression Analysis of the Chalcone Synthases (CHS) in <i>Osmanthus fragrans</i>

In the flavonoid biosynthesis pathway, Chalcone synthase (CHS) is involved in the formation of the pigment and has been shown to be a rate-limiting enzyme for the synthesis of flavonoids. In this study, a PCR approach was used to clone a Chalcone synthases cDNA from flower of sweet osmanthus “Chenghong Dangui” and it was designated as OfCHS (O. fragrans, CHS). The cDNA was 1383 bp long and a coding sequence (CDS) of 1173 bp encoding a polypeptide of 391 amino acids with an estimated molecular mass of 39.9 kDa. The theoretical isoelectric point was 6.23. Phylogenetic analysis demonstrated that OfCHS clustered with Olea europaea, Solenostemon scutellarioides, Perilla frutescens, Antirrhinum majus and Digitalis lanata. We also detected the expression of OfCHS in different tissues in “Dangui” and in two cultivars with varied coloration, “Zi Yingui” and “Chenghong Dangui” at different floral stages using quantitative real-time PCR. We observed that OfCHS transcript was higher in leaves than in petals in “Dangui”. The transcripts of OfCHS in “Zi Yingui” petals were higher than those in “Dangui” at three stages especially at xianyan stage and there was no significant difference between the two cultivars in the full flowering stage. “Chenghong Dangui” has a relatively high anthocyanin content compared to “Zi Yingui”. The relative amount of anthocyanin of “Chenghong Dangui” initially increases, and then decreases during the bloom period. However, the expression of CHS is the highest at the initial flowering stage. These data suggest that the OfCHS does not play a key role in the accumulation of total flavonoid in this cultivar. These data could contribute to explain the different accumulation of flavonoids in petals of the two cultivars.

The Genetic Relationship of Two New Colorful Sweet Osmanthus fragrans Cultivars Revealed by AFLP and SSR Markers

Osmanthus fragrans is well-known as an ornamental and agricultural plant noted for its unique fragrance.We investigated the genetic relationships between two new colorful O.fragrans cultivars,Ziyan Gongzhu and Qiannan Guifei,and their relationships with other O.fragrans using amplified fragment length polymorphism(AFLP)and simple sequence repeat(SSR),compared to the Changye Muxi species.55 AFLP primer pairs and 17 SSRs were screened;1103 and 45 amplification products were produced,of which 92.29%of the AFLP and 62.20%of the SSRs were polymorphic.At the population level,the number of effective alleles varied from 0.76 to 1.11,and the Shannon index(I)ranged from 0 to 0.11,indicating a narrow genetic base of O.fragrans cultivars.The genetic similarity varied from 0.61 to 0.80 between Ziyan Gongzhu and other O.fragrans cultivars,and 0.57 to 0.67 in the Qiannan Guifei and other cultivars.The unweighted pair group method with arithmetic mean(UPGMA)clustering revealed that Ziyan Gongzhu may belong to the O.fragrans Albus group.However,the Qiannan Guifei cultivar was clustered into a group,showing that it has great genetic variation.These results provide new molecular evidence for germplasm resources protection,and new cultivars for breeding.

Study on Prediction of the First Flowering Date of Osmanthus fragrans

[ Objective] The study aimed to forecast the first flowering date of Osmanthus fragrans. [ Method] Based on the data about flowering stage of O. fragrans in Guilin City from 1999 -2012, the correlation between meteorological factors and the first flowering date of O. fragrans was analyzed, and a forecast model for the first flowering date of O. fragrans was established. EResult] Among meteorological factors, precipitation, humidity, temperature and sunshine hours could obviously affect the first flowering date of O. fragrans, especially humidity in August, temperature and sunshine hours in September. The forecast model could exactly forecast the first flowering date in the partial early or too late year, and the av- erage error of date was 6 days. The factors influencing the first flowering date of O. fragrans are complex, and there is an artificial error in record data of the flowering stage, so using a single model to forecast the first flowering date have some disadvantages. When forecasting the first flower- ing date, we should consider all influencing factors to get a good forecast result. [ Conclusion] The research could provide a new method to forecast the first flowednq date of O. fraclrans.

Climatic Regionalization of Orange Osmanthus in Pucheng County Based on GIS

[Objectives]This study was conducted to scientifically plan orange osmanthus planting in Pucheng County to promote the development of agricultural economy.[Methods]Two factors,the total number of days with daily average temperature≥15℃from March to August and the total number of days with daily average temperature≥20℃in September were determined as the regionalization index factors using the weather data and geographic information data of Pucheng County,according to the 80%guarantee rate principle,the mean square error method and the actual growth law of orange osmanthus.Then,according to the weighted stack method,comprehensively considering the on-site inspection results and expert opinions,the suitability of orange osmanthus planting layout in Pucheng County was evaluated,and the GIS spatial interpolation technology was applied to complete the refined agroclimatic regionalization.[Results]The growth of Pucheng orange osmanthus has a great relationship with the thermal conditions,and it is clear that the suitable,sub-suitable and unsuitable areas for orange osmanthus planting in Pucheng County have certain applicability and maneuverability.[Conclusions]This study will play a scientific guiding role in the industrialization and development of orange osmanthus planting in Pucheng County.

Development Prospects of Osmanthus fragrans Mixed Wine Based on Liquor and Spirits

As a plant resource, Osmanthus fragrans has both medicinal and edible effects. This paper reviews the active components and health-care functions of O. fragrans, and introduces the current research situation of O. fragrans mixed wine. In addition, combined with the development of new products of plant-based flower wine, it analyzes the development prospects of O. fragrans mixed wine based on liquor and spirits.

A Study on Cold Resistance of Osmanthus Fragrans Cultivars in Kaifeng, China

Low winter temperature is generally recognized as the chief factor limiting the northward distribution of Osmanthus fragrans. O. fragrans has been cultivated in Kaifeng for nearly two decades, yet little is known regarding how well this plant has adapted to the city＇s cold winter. In a city-wide survey, we periodically examined O. fragrans leaves for visible symptoms of freeze damage, then measured leaf soluble sugar content, leaf electric conductivity, palisade layer thickness/leaf thickness ratio, and spongy layer thickness/leaf thickness ratio of several cultivars. The data thus collected were assessed to determine the cultivars＇ cold resistance levels. Our results indicate that the northward distribution of O. fragrans may be limited primarily by low spring temperatures rather than low winter temperatures. O. fragrans Sijigui and Huangchuan Jingui are the most cold resistant O. fragrans cultivars in Kaifeng, China.

A Comparative Study of the Translation of Reporting Verbs and Characterization between Conventional Translation and Self-Translation:A Case Study of Ah Xiao in Eileen Chang’s“Steamed Osmanthus”

Reporting verbs are not only an important means of presenting characters’speeches,but also an important means of characterization in narratives.The original and the two English versions of Eileen Chang’s novella“Steamed Osmanthus”are used as a corpus to conduct a comparative analysis focusing on the reporting verbs of the heroine Ah-Xiao.The analysis shows that the two translators-Eileen Chang herself and Simon Patton-have different styles.Patton favors the use of structuring verbs(especially“replied”)and neutral verbs in serial verb constructions while Chang tends to use neutral verbs glossed by an adverb,an adjective or a prepositional phrase.Descriptive verbs,especially prosodic verbs,also predominate in Chang’s version.As a result,both versions may have shaped a different Ah Xiao.

Sweet Osmanthus

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不同辐射剂量处理的盆栽桂花生长发育比较

为研究辐射对盆栽桂花(Osmanthus fragrans)生长发育的影响,以四季桂、金桂为供试材料,采用0、15、25、35 Gy 4种辐射剂量的60Co-γ射线对其进行辐射,统计桂花树新生芽数、新生叶数和株高,并测量叶长、叶宽和叶片质量,检测新生叶的过氧化物酶(POD)和过氧化氢酶(CAT)活性。结果表明,随着辐射剂量的增加,金桂的新生叶数、株高呈降低趋势,四季桂的新生叶数、新生芽数、CAT活性呈降低趋势。在一定的辐射剂量内,金桂的新生芽数、叶长、叶宽、叶片质量、POD活性、CAT活性呈升高趋势,四季桂则是株高、叶长、叶宽、叶片质量、POD活性呈升高趋势。综合来看,辐射对四季桂的影响更大,金桂的耐辐射能力更强。0~15 Gy的辐射剂量一定程度上促进桂花的生长发育。

Analysis of aroma-active compounds in three sweet osmanthus(Osmanthus fragrans) cultivars by GC-olfactometry and GC-MS

Objective：Aroma is the core factor in aromatherapy. Sensory evaluation of aromas differed among three sweet osmanthus （Osmanthus fragrans） cultivar groups. The purpose of this study was to investigate the aroma-active compounds responsible for these differences. Methods： Gas chromatography-olfactometry （GC-O） and GC-mass spectrometry （GC-MS） were used to analyze the aroma-active compounds and volatiles of creamy-white （‘Houban Yingui’, HBYG）, yellow （‘Liuye Jingui’, LYJG）, and orange （‘Gecheng Dangui’, GCDG） cultivars. Results：Seventeen aroma-active compounds were detected among 54 volatiles. trans-β-Ocimene, trans-β-ionone, and linalool, which were major volatiles, were identified as aroma-active, while cis-3-hexenyl butanoate,γ-terpinene, and hexyl butanoate were also aroma-active compounds, although their contents were low. Analysis of the odors was based on the sum of the modified frequency （MF） values of aroma-active compounds in different odor groups. HBYG contained more herb odors, contributed by cis-β-ocimene and trans-β-ocimene, while LYJG had more woody/violet/fruity odors released by trans-β-ionone, α-ionone, and hexyl butanoate. In GCDG, the more floral odors were the result of cis-linalool oxide, trans-linalool oxide, and linalool. Conclusions：Aroma-active compounds were not necessarily only the major volatiles：some volatiles with low content also contributed to aroma. The aroma differences among the three cultivars resulted from variation in the content of different odor groups and in the intensities of aroma-active compounds.