Based on non-targeted metabolomics for differential components screening of Rosae Chinensis Flos and Rosae Rugosae Flos and their quality evaluation

Background:Rosa chinensis Jacq.and Rosa rugosa Thunb.are not only of ornamental value,but also edible flowers and the flower buds have been listed in the Chinese Pharmacopoeia as traditional medicines.The two plants have some differences in efficacy,but the flower buds are easily confused for similar traits.In addition,large-scale cultivation of ornamental rose flowers may lead to a decrease in the effective components of medicinal roses.Therefore,it is necessary to study the chemical composition and make quality evaluation of Rosae Chinensis Flos(Yueji)and Rosae Rugosae Flos(Meigui).Methods:In this study,40 batches of samples including Meigui and Yueji from different regions in China were collected to establish high-performance liquid chromatography fingerprints.Then,the fingerprints data was analyzed using principal component analysis,hierarchical cluster analysis,and partial least squares discriminant analysis analysis chemometrics to obtain information on intergroup differences,and non-targeted metabolomic techniques were applied to identify and compare chemical compositions of samples which were chosen from groups with large differences.Differential compounds were screened by orthogonal partial least-squares discriminant analysis and S-plot,and finally multi-component quantification was performed to comprehensively evaluate the quality of Yueji and Meigui.Results:The similarity between the fingerprints of 40 batches roses and the reference print R was 0.73 to 0.93,indicating that there were similarities and differences between the samples.Through principal component analysis and hierarchical cluster analysis of fingerprints data,the samples from different origins and varieties were intuitively divided into four groups.Partial least-squares discriminant analysis analysis showed that Meigui and Yueji cluster into two categories and the model was reliable.A total of 89 compounds were identified by high resolution mass spectrometry,mainly were flavonoids and flavonoid glycosides,as well as phenolic acids.Eight differential components were screened out by orthogonal partial least-squares discriminant analysis and S-plot analysis.Quantitative analyses of the eight compounds,including gallic acid,ellagic acid,hyperoside,isoquercitrin,etc.,showed that Yueji was generally richer in phenolic acids and flavonoids than Meigui,and the quality of Yueji from Shandong and Hebei was better.It is worth noting that Xinjiang rose is rich in various components,which is worth focusing on more in-depth research.Conclusion:In this study,the fingerprints of Meigui and Yueji were established.The chemical components information of roses was further improved based on non-targeted metabolomics and mass spectrometry technology.At the same time,eight differential components of Meigui and Yueji were screened out and quantitatively analyzed.The research results provided a scientific basis for the quality control and rational development and utilization of Rosae Chinensis Flos and Rosae Rugosae Flos,and also laid a foundation for the study of their pharmacodynamic material basis.

Mechanism of Rosae Rugosae Flos flavonoids in the treatment of hyperlipidemia and optimization of extraction process based on network pharmacology

This study aims to identify a natural plant chemical with hypolipidemic effects that can be used to treat high cholesterol without adverse reactions.Through network pharmacology screening,it was found that Rosae Rugosae Flos(RF)flavonoids had potential therapeutic effects on hyperlipidemia and its mechanism of action was discussed.TCMSP and GeneCards databases were used to obtain active ingredients and disease targets.Venn diagrams were drawn to illustrate the findings.The interaction network diagram was created using Cytoscape 3.8.0 software.The PPI protein network was constructed using String.GO and KEGG enrichment analysis was performed using Metascape.The results revealed 2 active flavonoid ingredients and 60 potential targets in RF.The key targets,including CCL2,PPARG,and PPARA,were found to play a role in multiple pathways such as the AGE-RAGE signaling pathway,lipid and atherosclerosis,and cancer pathway in diabetic complications.The solvent extraction method was optimized for efficient flavonoid extraction based on network pharmacology prediction results.This was achieved through a single factor and orthogonal test,resulting in an optimum process with a reflux time of 1.5 h,a solid-liquid ratio of 1:13 g/mL,and an ethanol concentration of 50%.

Exploring the mechanism of Crocus sativus and Rosa rugosa for the treatment of coronary heart disease based on network pharmacology and molecular docking

Coronary atherosclerotic heart disease(CHD)is the main type of cardiovascular disease.The efficacy of Uyghur drug compound Saffron formula in CHD has been clinically proven.However,the underlying mechanism remains unclear.In this study,researchers investigated the active ingredients and mechanism of action of Crocus sativus and Rosa rugosa in the treatment of CHD by network pharmacology and molecular docking techniques,collected target information with the help of TCMSP,GEO,GeneCards,and other databases,constructed protein-protein interaction(PPI)network diagrams by STRING database,performed GO and KEGG pathway enrichment analysis on common targets,and finally molecularly docked the active ingredients with core targets.C.sativus-R.rugosa have a variety of polyphenol compounds,a total of 12 active ingredients,including quercetin and kaempferol,were screened.The first three targets intersected with the core targets of CHD as AKT1,TNF,and IL-1B.Enrichment results of KEGG pathway showed that C.sativus-R.rugosa against CHD involved atherosclerosis pathways.The molecular docking results showed that quercetin and kaempferol were well bound to the core targets,and it was speculated that these components might be the main active ingredients for the treatment of CHD.The potential mechanism of action of C.sativus-R.rugosa for the treatment of coronary heart disease was initially revealed.

‘紫枝’玫瑰(Rosa rugosa ‘Zi zhi’)开花过程花青素相关化合物及代谢途径分析

【目的】分析‘紫枝’玫瑰5个开花时期花瓣中的花青苷、类黄酮苷和类胡萝卜素的种类和含量,推定‘紫枝’玫瑰花的花青苷代谢途径,为探讨玫瑰花色的呈色机理和花色育种提供参考。【方法】以‘紫枝’玫瑰不同开花时期的花瓣为试材,用高效液相色谱(HPLC)和超高效液相色质联用分析法(UPLC-DAD-Q-TOF-MS)对其花青苷、类黄酮苷和类萝卜素进行结构推定和定量分析,结合化学反应过程推测‘紫枝’玫瑰花青苷代谢途径。【结果】在‘紫枝’玫瑰中鉴定出8种花青苷、16种类黄酮苷和β-胡萝卜素,没有检测到叶黄素;花青苷主要以芍药素、飞燕草素、矢车菊素和天竺葵素的双糖苷为主,四类花青苷都在花蕾期和初开期相对含量最高;检测到芍药苷的两种甲基化衍生物,没有发现飞燕草苷的甲基化衍生物;类黄酮苷以槲皮素和山萘酚的糖苷化、酰基化和甲基化的衍生物为主。定量分析显示,芍药苷和飞燕草苷占‘紫枝’玫瑰总花青苷含量的90%以上;芍药苷含量在‘紫枝’玫瑰开花过程中随花色变浅而降低,飞燕草苷在开花过程含量变化不大;芍药苷和飞燕草苷的比例(Pn/Dp)随花色变浅而降低。【结论】‘紫枝’玫瑰花中含有芍药素、飞燕草素、矢车菊素和天竺葵素,这四类花青素在半开期之前完成全部积累,盛开期后只降解不合成。芍药素是形成‘紫枝’玫瑰花色的主要成分。‘紫枝’玫瑰的花青苷代谢途径中,甲基酶(Rr AOMT)的催化作用有底物特异性。

A study on JA-and BTH-induced resistance of Rosa rugosa‘Plena’to powdery mildew(Sphaerotheca pannosa)

Different concentrations of jasmonic acid(JA)and benzothiadiazole(BTH) were sprayed on 2-year-old Rosa rugosa‘Plena’ seedlings. The induced resistance of JA and BTH to Sphaerotheca pannosa(Wallr.) and the changes of their related physiological indices were investigated. Results showed that JA and BTH treatments had inhibitory impacts on S. pannosa infection. The optimal concentration of JA and BTH was 0.5 mmol/L for the disease-resistance induction of the leaves, its inductive effect was up to 66.36% for BTH and 54.49% for JA. Our results confirmed that exogenous JA and BTH significantly improved R. rugose ‘Plena’ resistance to S. pannosa. When treated with JA and BTH, activities of the three defense enzymes(POD, PPO, and PAL) increased significantly.Contents of total phenolics, flavonoids, and lignin also increased significantly. It is inferred from these results that exogenous JA and BTH could improve the resistance of R.rugose ‘Plena’ to S. pannosa through enhancing activities of the defensive enzymes and accumulation of secondary metabolites in the leaves.

Genetic diversity of the endangered species Rosa rugosa Thunb.in China and implications for conservation strategies

Rosa rugosa Thunb. is one of the dominant and important shrub species in estuary dunes and shingle beaches of northern China. However, its area of distribution, the number of populations, and the size of each population have decreased rapidly in the past two decades because of habitat degradation and loss. Random amplified polymorphic DNA markers were used to determine the genetic diversity of four remaining large natural populations of R. rugosa and to discuss an effective conservation strategy for this endangered species in China. High genetic variations were detected in R. rugosa populations in China. The mean percentage of polymorphic loci （P%） within four local populations was 57.99%, with the P% of the total population being 75.30%. Mean Shannon＇s information index （H0） was 0.2826, whereas total Ho was 0.3513. The genetic differentiation among populations was 0.1878, which indicates that most genetic diversity occurs within populations. Population Tumenjiang （TMJ） showed the highest genetic diversity （P% = 66.27%; H0 = 0.3117） and contained two exclusive bands. Population Changshandao （CSD） showed higher genetic diversity （P% =59.04%; H0 = 0.3065）. Populations TMJ and CSD contained 95.33% and 99.33%, respectively, of loci with moderate to high frequency （P〉0.05） of the total population. These results indicate that populations TMJ and CSD should be given priority for in situ conservation and regarded as seed or propagule sources for ex situ conservation. The results of the present study also suggest that R. rugosa in China has become endangered as a result of human actions rather than genetic depression of populations; thus, human interference should be absolutely forbidden in R. rugosa habitats.

Effect of jasmonate treatments on leaves of Rosa rugosa‘Plena’and detoxification enzymes and feeding of adult Monolepta hieroglyphica

To study the effect of jasmonates(JAs)on the resistance of economic forest plants to insects,R osa rugosa‘Plena'leaves were treated with 1 mmol/L jasmonic acid(JA),methyl jasmonate(MeJA)and Z-jasmone,then the content of tannin and total phenol in leaves and the feeding area of Monolepta hieroglyphica adults on leaves were measured.Changes in the activities of detoxification enzymes in adult M.hieroglyphica that had fed on leaves treated with JAs were also studied.Tannin and total phenol levels in leaves increased significantly after treatment with JAs,and tannin level was 1.36–1.55-fold higher than in the control after treatment with 1 mmol/L MeJA.The total content of phenol in leaves treated with 1.0 mmol/L Z-jasmone increased by 1.33–2.20-fold compared with those of the control.The activities of detoxification enzymes in adults were inhibited to differing degrees:activity of alkaline phosphatase(AKP)first increased,then decreased;the activities of acid phosphatase(ACP),glutathione S-transferases(GSTs)and carboxylesterase(CarE)following treatment with 1 mmol/L MeJA were significantly reduced and were 22%–31%,11%–26%,and 11%–31%lower than those of the control,respectively.Moreover,the feeding area of adult M.hieroglyphica on the leaves treated with JAs was significantly reduced(P<0.05).The feeding area of economic forest R.rugosa‘Plena'leaves treated with 1 mmol/L MeJA decreased by 17%–43%compared with that of the control.Moreover,the decrease in the adult M.hieroglyphica feeding area was highly positively correlated with the content of tannin and positively correlated with the contents of total phenol of economic forest R.rugosa‘Plena'leaves.The reduced feeding area of adult M.hieroglyphica was highly negatively correlated with the activities of AKP and ACP and negatively correlated with those of the GSTs.In conclusion,the use of 1 mmol/L MeJA can noticeably decrease the deleterious effects of adult M.hieroglyphica.

Cloning and Expression Analysis of <i>RrGT2</i>Gene Related to Anthocyanin Biosynthesis in <i>Rosa rugosa</i>

At present, the research about flower color of Rosa rugosa is a very inno-vative and practical study. Glycosylation modification fulfills an important role in increasing the stability and solubility of anthocyanin in plants. In this study, based on the transcriptional database of R. rugosa, a gene with full length cDNA of 1422bp, encoding 473 amino acids, designated as RrGT2, were isolated from flowers of R. rugosa ‘Zizhi’ and then functionally characterized. According to online software prediction, the molecular formula of the protein encoded by the RrGT2 gene is C2334H3628N602O711S18, the relative molecular mass is 52,075.17 Da, and the theoretical isoelectric point is pI = 4.76. The result of the RrGT2 protein 3D model construction showed that it had the highest homology with the UDP-glycosyltransferase 74F2 protein model in the database (39.53%). Sequence alignments with the NCBI database showed that the RrGT2 protein is a member of the GTB superfamily. Homology analysis revealed that the coding regions of RrGT2 was highly specific among different species, but still had typical conserved amino acid residues called PSPG that are crucial for RrGT2 enzyme activity. RrGT2 transcripts were detected in five flowering stages and seven tissues of R. rugosa ‘Zizhi’, R. rugosa ‘Fenzizhi’ and R. rugosa ‘Baizizhi’, and their expression patterns corresponded with the accumulation of antho-cyanins. Therefore, we speculated that glycosylation of RrGT2 plays a crucial role in anthocyanin biosynthesis in R. rugosa.

Component Analysis of Anthocyanins in Petals at Different Flowering Stages of Three <i>Rosa rugosa</i>Hybrid Cultivars

The species and contents of anthocyanins in plant petals can make plants appear pink, red, violet and blue, etc., and play a major role in the coloration of plants. In this study, the species and contents of anthocyanins in the petals of three R. rugosa hybrid cultivars, R. rugosa “Zizhi”, R. rugosa “Fenzizhi” and R. rugosa “Baizizhi”, were analyzed, and the direct cause of the differences in flower color of three R. rugosa hybrid cultivars was inferred. This paper provides a reference for the coloration mechanism and flower color breeding of R. rugosa. The specific methods are as follows: the petals of five flowering stages of three R. rugosa hybrid cultivars were used as materials, and the types and contents of anthocyanins contained in them were qualitatively and quantitatively analyzed by high performance liquid chromatography (HPLC). The same six kinds of anthocyanins were identified in R. rugosa “Zizhi” and R. rugosa “Fenzizhi”, mainly based on the diglycoside of paeoniflorin and cyanidin. The relative contents of the two anthocyanins were higher at budding stage and initial opening stage. In the different flowering stages of R. rugosa “Zizhi”, the content of Pn3G5G was up to 4280.84 ± 20.82 μg&middot;g-1, and the content of Cy3G5G was up to 789.41 ± 1.21 μg&middot;g-1. In R. rugosa “Fenzizhi”, the highest content of Pn3G5G reached 1293.50 ± 17.64 μg&middot;g-1, and the content of Cy3G5G was up to 358.86 ± 3.94 μg&middot;g-1. It could be speculated that the difference in the contents of Pn3G5G and Cy3G5G was the main reason for the difference in coloration between the petals of R. rugosa “Zizhi” and R. rugosa “Fenzizhi”. A total of five species of anthocyanins were identified in R. rugosa “Baizizhi” and their contents were relatively low. Compared with R. rugosa “Zizhi” and R. rugosa “Fenzizhi”, the presence of Cy3G was not detected. Therefore, we speculated that the two reasons above might be responsible for the visual white flowers of R. rugosa “Baizizhi”.

Cloning and Expression of Anthocyanin Biosynthesis Related Gene RrMYB6 in <i>Rosa rugosa</i>

R2R3-MYB transcription factor plays an important role in plant anthocyanin synthesis. Based on the transcriptional database of Rosa rugosa, one MYB transcription factor related to floral color, RrMYB6, was cloned. By using bioinformatics analysis method, cloning MYB gene and analyzing its function in anthocyanin biosynthesis regulation, we hope to lay a solid foundation for new color variety breeding of R. rugosa. Using the R. rugosa “Zi zhi” as the material, we obtained the total length of cDNA of RrMYB6 by RT-PCR and RACE. By analyzing its bioinformatics, we found that the formula of the protein was C1491H2368N452O470S17, molecular weight was 34690.97 Da, the theoretical pI was 8.74. In addition, it belonged to unstable protein with an unstable index at 50.59, and it was also a hydrophilic protein with the total average hydrophobic index at -0.847. In the secondary structure of RrMYB6 protein, the Alpha helix accounted for 32.35%, random coil was 47.39%, extended strand was 11.11%, and beta turn was 9.15%. The sequence analysis showed that RrMYB6 had a typical R2R3-MYB domain and bHLH binding domain, and it also had an N1, C1, C2 inhibitory motif, belonging to the Sg4 subfamily MYB protein. What’s more, evolutionary analysis indicated that the RrMYB6 protein was closely related with the MYB protein in Rosacea family, while it was far from those in other families. The expression analysis showed that RrMYB6 protein decreased with the color of petals deeping, and its expression was the lowest in the petals while the highest in stamens. According to the above results, it was speculated that RrMYB6 was involved in regulating the anthocyanin synthesis of R. rugosa, which belonged to negative regulatory mechanism.

Cloning and Bioinformatics Analysis of Rosa rugosa β-1,3-Glucanase Gene (RrGlu)

In order to reveal which role the callose played in R. rugosa pollination incompatibility, the full-length cDNA sequence of β-1,3-glucanase gene was cloned for the first time from the stylus of Rosa rugosa “Tanghong” with RT-PCR and RACE methods and named as RrGlu. The full-length cDNA is 1380 bp with an open reading frame of 1041 bp, encoding 346 amino acids. The derived protein has a molecular weight of 37.85 kD, a calculated pI of 9.12, a pfam00332 conserved domain at position 36 - 345, and belongs to glycosyl hydrolase family 17. The derived protein is a hydrophilic protein secreted into the vacuole. There is a signal peptide cleavage site at position 34 - 35, a transmembrane domain at position 13 - 32, six Ser phosphorylation sites, three Thr phosphorylation sites, three Tyr phosphorylation sites, one N-glycosylation site, and five O-glycosylation sites. There are 31.50% α-helixes, 30.92% random coil, 25.14% extended peptide chain, and 12.43% β-corner structure. This protein and the Glu protein from eight other species, including Prunus persica, share a sequence homology of greater than 72%;all of the proteins contain a pfam00332 conserved domain and a β-1,3-glucanase active center sequence (LIVM)-X-(LIVMFYW)3-(STAG)-E-(ST)-G-W-P-(ST)-X-G. Furthermore, their phylogenetic relationships are consistent with their traditional classifications. These results were meaningful to reveal the molecular mechanism of R. rugosa pollination incompatibility and improve the theory and techniques of breeding ornamental R. rugosa.

Clonging and Analysis of RrF3’ H in Rosa rugosa

Rosa rugosa is an important garden ornamental plant which belongs to the genus Rosa of the family Rosaceae. The current wild and cultivated R. rugosa are mostly purple, pink, a small amount of white, but lack of yellow, orange and so on. Flavonoids 3’-hydroxylase belongs to CYP75B subfamily of cytochrome P450, and is an essential enzyme in anthocyanins synthesis. In this experiment, RrF3’H gene was cloned from the petal of Rosa rugosa ‘Hunchun’ using RT-PCR, and bioinformatics analysis was performed. The RrF3’H gene’s full length of opening reading frame was 1687 bp, encoding 510 amino acids. The formulas of proteins encoded by RrF3’H were C2666H4149N699O734S24. The derived protein had a molecular weight of 58,506.95 Da. The aliphatic index was 90.94. It belongs to unstable hydrophilic protein. The protein consists of 46.76% α-helix, 31.04% random coil, 7.66% β-corner and 14.54% extended strand. The protein contains 21 Ser phosphorylation sites, 12 Thr phosphorylation sites, and 2 Tyr phosphorylation sites. The protein contained two O-glycosylation sites, located at positions 98 and 263 of the amino acid sequence respectively. The protein has a signal peptide site and a transmembrane structure. In addition, by comparing the expression levels of RrF3’H, we found RrF3’H was positively correlated with the depth of flower color.

Cloning and Expression of One Anthocyanin-Related R2R3-MYB Gene in <i>Rosa rugosa</i>

Based on the transcriptome of Rosa rugosa, one anthocyanin-promoting R2R3-MYB gene, RrMYB10.1 (Accession Nos:MH717244), was cloned from the petals of Rosa rugosa ‘Zizhi’. Sequence analysis results showed that RrMYB10.1 had a full length opening reading frame of 747bp, encoding 249 amino acids. Sequence analysis revealed that RrMYB10.1 contained the conserved R2R3-MYB domain, two atypical anthocyanin-promoting motifs and a conserved amino acid signature for the interaction with bHLH protein. The results of phylogenic tree revealed that RrMYB10.1 showed high homology with other anthocyanin-promoting proteins in Rosacea, and sharing the highest identity (98.39%) with RhMYB10. RT-PCR results showed that RrMYB10.1 was mainly expressed in petals among various tissues and expressed significantly higher in petals in bud stage than in opening period. To sum up, these results showed that RrMYN10.1 may play a key role in regulating anthocyanin concentration, thus providing a certain foundation on regulating flower color formation in Rosa rugosa.

Overexpression of the <i>Rosa rugosa RrGT1</i>Gene Induces Anthocyanin Accumulation in Tobacco

Rosa rugosa has always been an important plant in landscape application, and the improvements and innovations about its flower color are particularly important. Glycosylation modification fulfills an important role in increasing the stability and solubility of anthocyanin in plants. In this study, based on the transcriptional database of R. rugosa, a gene with full length cDNA of 1161 bp, encoding 386 amino acids, designated as RrGT1, were isolated from flowers of R. rugosa “Zizhi” and then functionally characterized. Sequence alignments with the NCBI database show that the RrGT1 protein is a member of the GTB superfamily and has typical conserved amino acid residues called PSPG that are crucial for RrGT1 enzyme activity. RrGT1 transcripts were detected in five flowering stages and seven tissues of R. rugosa “Zizhi” and their expression patterns corresponded with the accumulation of anthocyanins. Additionally, the in vivo function of RrGT1 was investigated via its overexpression in tobacco. Transgenic tobacco plants expressing RrGT1 induced anthocyanin accumulation in flowers, indicating that RrGT1 could encode a functional glycosyltransferase (GT) protein for anthocyanin biosynthesis and could function in other species. Therefore, we speculated that glycosylation of RrGT1 played a crucial role in anthocyanin biosynthesis in R. rugosa.

Cloning and Bioinformatics Analysis of <i>Rosa rugosa</i>TFL1 Gene (<i>RrTFL1</i>)

In order to determine if the TFL1 is related with the continuous flowering phenotype of wild Rosa rugosa from Muping, the full-length cDNA sequence of TFL1 Gene was cloned for the first time from the flower buds of wild Rosa rugosafrom Muping with RT-PCR and RACE methods and named as RrTFL1. The full-length cDNA is 973 bp with an open reading frame of 519 bp, encoding 172 amino acids. The derived protein has a molecular weight of 19.48 kD, a calculated pI of 9.13, a c100227 conserved domain at position 1-172, and belongs to PEBP family. The derived protein is a Hydrophilic protein secreted into the cytoplasmic. There is no transmembrane domain and no signal peptide cleavage site, five Ser phosphorylation sites, seven Thr phosphorylation sites, three Tyr phosphorylation sites, one O-glycosylation site, and no N-glycosylation sites. There are 24.42% α-helixes, 36.63% random coil, 27.91% extended peptide chain, and 11.05% β-corner structure. This protein and the TFL1 protein from Rosaceae plants, including Rosa chinensis, share a sequence homology of 87% - 96%. All of the proteins contain a c100227 conserved domain, two highly conserved modules D-P-D-x-P, G-x-H-R, and two functional sites His, Asp. Furthermore, their phylogenetic relationships are consistent with their traditional classifications. These results not only laid a foundation for further researching the expression and function of RrTFL1, but also cultivating new varieties of R. rugosawhich can flower continuously by gene engineering.

Cloning and Expression Analysis of <i>RrGT1</i>Gene Related to Anthocyanin Biosynthesis in <i>Rosa rugosa</i>

Glycosylation modification fulfills an important role in increasing the stability and solubility of anthocyanin in plants. In this study, based on the transcriptional database of R. rugosa, a gene with full length cDNA of 1161 bp, encoding 386 amino acids, designated as RrGT1, was isolated from flowers of R. rugosa ‘Zizhi’ and then functionally characterized. According to online software prediction, the molecular formula of the protein encoded by the RrGT1 gene is C1879H2964N494O556S14, the relative molecular mass is 41,820.02 Da, and the theoretical isoelectric point is pI = 5.03. The result of the RrGT1 protein 3D model construction showed that it had the highest homology with the UDP-glucose: anthocyanidin 3-O-glucosyltransferase protein model in the database (47.01%). Sequence alignments with the NCBI database showed that the RrGT1 protein is a member of the GTB superfamily. Homology analysis revealed that the coding regions of RrGT1 was highly specific among different species, but still had typical conserved amino acid residues called PSPG that are crucial for RrGT1 enzyme activity. RrGT1 transcripts were detected in five flowering stages and seven tissues of R. rugosa ‘Zizhi’, R. rugosa ‘Fenzizhi’ and R. rugosa ‘Baizizhi’, and their expression patterns corresponded with the accumulation of anthocyanins. Therefore, we speculated that glycosylation of RrGT1 plays a crucial role in anthocyanin biosynthesis in R. rugosa.

Study on Cultivation Technology of Rosa rugosa Thunb.

Rosa rugosa Thunb. is a traditional medicinal and food plant in Hotan region,which has important medicinal and economic value. In recent years,the planting area in Hotan region has been expanding. This paper described the cultivation techniques,planting models and field management,pest control,harvesting and processing of R. rugosa. Based on the analysis of the advantages and cultivation techniques of developing the rose industry in Hotan region,the paper discussed how to accelerate the development of rose cultivation and deep processing industry in Hotan region.

Cloning and Bioinformatics Analysis of <i>Rosa rugose S</i>Locus F-Box Gene (<i>RrSLF</i>)

In order to reveal the phenomenon of R. rugose pollination incompatibility, the full-length cDNA sequence of S Locus F-box Gene was cloned for the first time from the pollen of Rosa rugose “Zilong wochi” with RT-PCR and RACE methods and named as RrSLF. The full-length cDNA is 1236 bp with an open reading frame of 1122 bp, encoding 343 amino acids. The derived protein has a molecular weight of 43.7 kD, a calculated pI of 6.24, an F-box conserved domain at position 343 - 741, and belongs to F-box family. The derived protein is a Hydrophobicity protein secreted into the cytoplasm. There is no transmembrane domain and no signal peptide cleavage site, twenty-one Ser phosphorylation sites, seven Thr phosphorylation sites, seven Tyr phosphorylation sites, two N-glycosylation sites, and no O-glycosylation sites. There are 22.25% α-helixes, 31.37% random coil, 32.17% extended peptide chain, and 14.21% β-corner structure. This protein and the SFB/SLF protein from Rosaceae Prunus fruit, including Prunus speciose, share a sequence homology of 59% - 61%;all of the proteins contain an F-box conserved domain, two hypervariable regions HVa, HVb, and two variable regions V1, V2. Furthermore, their phylogenetic relationships are consistent with their traditional classifications. These results were meaningful to reveal the molecular mechanism of Rosa rugose pollination incompatibility and improve the theory and techniques of breeding ornamental Rosa rugose.

三个玫瑰品种花色物质的代谢组学分析

玫瑰(Rosa rugosa)具有很高的观赏价值和商业价值,但其花色比较单一,限制了玫瑰的开发利用及其在园林造景中的应用。为了探究‘苦水玫瑰’‘墨红玫瑰’和‘保加利亚白玫瑰’3个不同品种玫瑰的呈色物质,该研究利用超高效液相色谱-四级杆-飞行时间质谱(UPLC-Q-TOF-MS)联用检测花瓣类黄酮的种类和含量,通过KEGG数据库对差异代谢物进行富集分析,筛选出关键代谢物,并分析与花色表型值的相关性。结果表明:(1)在3个不同色系玫瑰花瓣中共检测到58种代谢物,其中花青素只有一种为矢车菊-3-O葡萄糖苷,约占30.45%。(2)K-means聚类分析表明,共有12种关键代谢物注释到KEGG代谢通路中,其中乔松素和杨梅黄酮是决定‘苦水玫瑰’和‘墨红玫瑰’花色呈红色的主要物质,圣草酚、木犀草素和山萘酚是决定‘保加利亚白玫瑰’花色呈白色的主要物质。该研究结果可为具有特定颜色玫瑰的育种提供理论依据,并促进玫瑰在园林绿化中的应用。

玫瑰花HPLC指纹图谱研究

目的建立玫瑰花HPLC指纹图谱,为其质量评价提供参考。方法采用HPLC,色谱柱为COSMOSIL 5C18-MS-Ⅱ(4.6 mm×250 mm,5μm),以2.5%乙腈+0.1%甲酸+水(A)-乙腈+0.1%甲酸(B)为流动相,梯度洗脱,柱温40℃,流速0.5 mL/min,检测波长350 nm。采用《中药色谱指纹图谱相似度评价系统》(2012版)对13批样品进行相似度评价,通过液相色谱-质谱(LC-MS)技术进行定性分析,并结合聚类分析、主成分分析和正交偏最小二乘-判别分析对玫瑰花进行整体质量评价。结果建立了玫瑰花HPLC指纹图谱共有模式,13批样品相似度较好,初步鉴定指认其中9个共有峰;化学模式识别将13批样品聚为3类。结论本研究建立的玫瑰花指纹图谱结合化学模式识别方法灵敏度高、专属性强,可为制定玫瑰花的质量评价标准提供依据。