Changes of Relevant Indexes of Photosynthetic Pigment and Nitrogen Metabolism in Leaves of Chrysanthemum morifolium Ramat. ‘Huaibai' after TAV-free

[ Objective ] The paper was to explore effects of Tomato aspermy virus （TAV） -free on chlorophyll content and nitrogen metabolism of Chrysanthemum morifolium Ramat. ＇Huaibai＇, in order to provide a theoretical basis for field popularization and application of TAV-free ＇ Huai bai＇ plantlets. [ Method] With previously acquired TAV-free test-tube plantlets of ＇ Huaibai＇ as the experimental materials and its ordinary test-tube plantlets as the control （CK）, chlorophyll content and some indexes related to nitrogen metabolism in leaves were determined during subculture. [ Result] During subculture, the contents of chlorophyll a and chlorophyll b in leaves of CK and TAV-free ＇ Huaibai＇ test-tube plantlets increased with the subculture days. Both realized the largest increase at the 22＇h day of subculture, and their increase amplitudes were 5.24% and 41.69%, respectively. Compared with CK, the chlorophyll a and chlorophyll b contents of TAV-free test-tube plantlets increased at the same time point of subculture, with significant difference （P 〈0.05） or extremely significant difference （P 〈0.01 ）. Further- more, the nitrate reductase （NR） activity, free proline and soluble protein contents of TAV-free ＇ Huaibai＇ were all significantly higher than those of CK, and the highest increase amplitudes were 27.99% （ subculture for 22 d）, 28.10% （ subculture for 22 d） and 5.89% （ subculture for 38 d）, respectively. [ Conclusion] TAV-free could effectively improve chlorophyll content and nitrogen metabolism level of ＇ Huaibai＇ , and was conducive to its growth and development.

B Class Floral Homeotic Genes are Involved in the Petal Identity and Flower Meristem Determinations in Chrysanthemum morifolium

Chrysanthemum morifolium,an ornamental crop with diverse forms of inflorescence,is a good model for studying flower development in Asteraceae.However,the genetic background is complex and the mechanisms of regulating flower development are still unclear.Here,we identified two natural mutant lines of chrysanthemum and named them M1 and M2 according to the severity of the phenotype.Both lines showed defects in petal identity,and the petals of the M1 line had a mild phenotype:partially loss of petal identity and conversion of petals into green,leaf-like organs.The M2 line had severe phenotypes:in addition to severe petal defects,secondary inflorescences were produced in the capitulum to replace the normal ray and disc florets,which indicated a transformation of a flower meristem into an inflorescence meristem.Transcriptome sequencing of WT and M2 inflorescences was performed and found altered expression of floral organ development A,B and E class genes,where B and E class genes were significantly down-regulated.qRT-PCR analysis in both M1 and M2 lines revealed that the expression of three chrysanthemum class B genes CmAP3.1,CmAP3.2 and CmPI,was negatively correlated with phenotypic severity.This suggests that class B genes in chrysanthemum not only have conserved functions in determining petal identity but also were involved in the determinacy of the flower meristem.This study provides insights into the functions of class B genes in flower development,and is informative for dissecting the molecular mechanisms of flower development in chrysanthemum.

Optimization of Extraction Process of Polyphenols from Chrysanthemum morifolium and the Development of Chrysanthemum Rice Wine

[Objectives]This study was conducted to provide a theoretical basis for the extraction and comprehensive utilization of functional active components of Chrysanthemum morifolium polyphenols.[Methods]With C.morifolium as a raw material,polyphenols were extracted by water extraction.The extraction process of polyphenols were optimized by single factor tests on solid-liquid ratio,extracting time and times.Old rice wine selected as the base wine was added with C.morifolium polyphenol extract,honey,citric acid and other auxiliary materials to prepare a kind of chrysanthemum old rice wine.Through sensory analysis combined with fuzzy mathematics comprehensive evaluation method,the optimal formula of chrysanthemum old rice wine was determined,and the corresponding physicochemical indicators of the obtained chrysanthemum old rice wine were tested.[Results]The optimal soaking process for C.morifolium was as follows:extraction time of 20 min,a solid-liquid ratio of C.morifolium to warm water at 1:40,extraction times of 3 times.The physical and chemical indicators of the chrysanthemum wine obtained from the above optimal formula were as follows:sugar content 5%,ethanol content 11%,and pH value 4.04.The chrysanthemum old rice wine obtained was clear and transparent,and the fragrance of chrysanthemum was coordinated with the mellow aroma of rice wine.The taste was refreshing and suitable for the vast majority of people to drink.[Conclusions]The new type of chrysanthemum old rice wine combines the polyphenolic active substances in C.morifolium with the low ethanol content of old rice wine,which not only improves the utilization rate of C.morifolium,but also strengthens the health function of old rice wine products,improves the quality of old rice wine,and promotes the healthy and rapid development of the old rice wine industry.

Flavonoid extracts from chrysanthemum with appropriate anthocyanins turn blue when exposed to iron ions

Although some species that accumulate only cyanidin(Cy)in nature can produce blue flowers through iron ions,there has been no evidence of blue chrysanthemums being generated in this manner.This study revealed that flavonoid extracts from the ray florets of the chrysanthemum cultivar‘Wandai Fengguang’turned blue when exposed to Fe^(3+).Samples that could turn blue were labeled as CB(Cy-determined blue flowers),while samples that did not turn blue were labeled as CN(Cy-determined non-blue flowers).After a series of experiments,a stable screening system was established using flavonoid extracts containing NaAc buffer at pH 5.5 and a total anthocyanin concentration(TAC)of 30 μmol·L^(-1),and the addition of Fe^(3+)from 0 to 0.25 μmol·L^(-1)allowed for the selection of five CB samples from 39 chrysanthemum cultivars.All five CB samples exhibited flower color phenotypes that belonged to Cluster-I with redness(a*)values ranging from 29.03 to 45.99,yellowness(b*)values from-11.31 to 3.77,and brightness(L*)values from 29.07 to 45.99.Additionally,the ratio of TAC to total luteolin concentration(TLC)was found to be a critical factor for distinguishing between CB and CN samples.To realize the desired blue hue in the flavonoid extracts with the participation of Fe^(3+),a TAC to TLC ratio of 2.25 and above is required.Moreover,the protoplasts and ray florets of CB samples that turned blue with the involvement of Fe^(2+)showed great potential for cultivating blue chrysanthemums through ferric-anthocyanin chelate.Overall,this study reveals that blue flowers can be cultivated through the increase in the iron ion concentration,combined with the accumulation of Cy.

FLOWERING LOCUS T(FT)in Photosensitive Type Chrysanthemum Accelerates Flowering in Arabidopsis

FLOWERING LOCUS T(FT)is an important factor for integrating flowering signals through the photoperiod pathway,which significantly promotes flowering.Most chrysanthemum varieties are short-day plants,sensitive to the photoperiod,which limits their ornamental and annual production supply.In this study,we aim to analyze the nutrients and flowering-related genes of chrysanthemums with different photoperiod types and to clone and verify the function of the flowering-related gene CmFT.We found that the formation of floral buds requires the accumulation of starch while consuming soluble sugars and the expression patterns of flowering-related genes GIGANTEA(GI),CONSTANS(CO),and FT in C.morifolium‘Zilian’and C.morifolium‘Zihongtuogui’had a certain synchronization during floral buds differentiation according to our quantitative validation,and the expression levels of CmGI,CmCO and CmFT in C.morifolium‘Zihongtuogui’were higher than those in C.morifolium.‘Zilian’in the later stage of differentiation.CmFT was cloned from photosensitive chrysanthemums-C.morifolium‘Zihongtuogui’and polypeptide alignment and phylogenetic analysis showed that CmFT was clustered in FT-like subfamily.In further functional verification,we obtained two Arabidopsis transgenic lines.Our results showed that CmFT transgenic ft mutant lines can significantly accelerate flowering in Arabidopsis.Thus,we can initially confirm that CmFT plays an important role in promoting flowering,which may be the key reason for the photosensitivity of C.morifolium‘Zihongtuogui’.Overall,the results of this study are of great importance in revealing the flowering mechanism of different photoperiod types of chrysanthemums.

Effect of Different Arbuscular Mycorrhizal Fungi Strains on Mineral Nutrition and Antioxidant Enzymes of Chrysanthemum morifolium

[Objective] The purpose was to study the effects of different arbuscular mycorrhizal fungi strains（AMF）on the mineral nutrition and antioxidant enzymes of Chrysanthemum morifolium.[Method] The pot experiment was conducted in the greenhouse,C.morifolium ＇Jinba＇ was inoculated with five kinds of AMF,N,P,K,malondial dehyde（MDA）content,as well as the superoxide dismutase（SOD）,peroxidase（POD） and catalase（CAT） activity in roots,leaves and petals of C.morifolium were measured at seedling and flowering stages.[Result] The G.i,G.e and G.m treatments could promote mineral nutrient absorption,increase N,P,K content in roots,leaves and petals of C.morifolium compared with the control without inoculation.The G.d、G.e and G.m treatments could significantly reduce MDA content in roots and petals,thus alleviating membrane permeability and lipid peroxidation.The G.i treatments could also improve the SOD,POD and CAT activities of C.morifolium,thereby increasing the capability of scavenging oxygen free radicals.[Conclusion] According to the comprehensive analysis,G.i was screened out as the best strain to improve mineral nutrition and antioxidant enzyme activities of C.morifolium.

Two New Flavonoid Glycosides from Chrysanthemum morifolium

Two new flavonoid glycosides were isolated from the flowering heads of Chrysanthemum morifolium. Their structures were determined to be luteolin 4＇-methoxy-7- O-（6＂-O-acetyl）-β-D-glucopyranoside （1） and acacetin 7-O-（3＂-O-acetyl）-β-D-glucopyranoside （2） by means of 1H and 13C NMR spectroscopic analysis, including 2D NMR technique.

Protection of Intellectual Property and Authenticity of Chinese Herbal Medicine Chrysanthemum morifolium

This paper first introduced Chrysanthemum morifolium Ramat. and its medicinal value. Then, it studied agricultural intellectual property resources such as geographical indications, trademarks, patents, new plant varieties, biological genetic resources, and traditional knowledge related to C. morifolium . Finally, it summarized poverty-stricken areas ecologically suitable for planting C. morifolium , analyzed the GAP certification of C. morifolium , pollution-free agricultural product certification, green food certification, and organic food certification, and discussed the strategies for C. morifolium intellectual property protection and authenticity protection in the context of TCM development strategy and rural revitalization strategy.

Effects of Different Treatment Methods on Virus Elimination of Chrysanthemum morifolium 'Shenma'

[Objective]The paper was to explore the virus elimination technique of Chrysanthemum morifolium′Shenma′.[Method]With C.morifolium ′Shenma′as the test materials,the virus elimination effects of shoot-tip treatment,shoot-tip with virazole treatment and shoot-tip with heat treatment were used to study the effect of virus elimination in the paper.The shoot-tips with the lengths of 0.2-0.3,0.4-0.5,0.6-0.8 and 0.9-1.0 mm were cultured by the shoot-tip treatment.A randomized complete blocks design was conducted to screen the best shoot-tip length and virazole concentration by shoot-tip with virazole treatment.The test-tube seedling were cultured by the method of variable temperature at day and night and stepwise temperature rising.ELISA test was used to identify the elimination efficiencies.[Result]The shoot-tip with the length of 0.4-0.5 mm was the best in shoot-tip treatment;the combination of shoot-tip length of 0.4-0.5mm and virazole mass concentration of 10 mg/L was the best in shoot-tip with virazole treatment;the shoot-tip with the length of 0.4-0.5 mm stripped after heat treatment for 45d was the best in shoot-tip with heat treatment.[Conclusion]The study provides an effective way for virus elimination in C. morifolium′Shenma′.

Target prediction of Chrysanthemum morifolium against hepatocellular carcinoma

A novel computational system was used to decipher the targets and mechanisms of Chrysanthemum morifolium against hepatocellular carcinoma.The putative target profile of Chrysanthemum morifolium against hepatocellular carcinoma was identified from Traditional Chinese Medicine Systems Pharmacology and DrugBank.Next,identification of the protein-protein interaction network of Chrysanthemum morifolium against hepatocellular carcinoma targets,and the identification of differentially expressed genes in hepatocellular carcinoma,relied on data from the NCBI database.Finally,Kyoto Encyclopedia of Genes and Genomes and Gene Ontology were used to analyze the common targets.Forty-eight active compounds in Chrysanthemum morifolium were identified by Traditional Chinese Medicine Systems Pharmacology and DrugBank,and 71 differentially expressed genes were identified in the GEO database.Further,10 core target proteins were selected,including CYP1A1,ADRB2,ADRA1B,PGR,MAOB,SLC6A4,GABRA1,MAOA,NOS3 and PTGS2.These target genes were also found to be associated with pathways involved in adrenergic receptor activity and catecholamine binding.

Hemmung der exoerythrozytaren Entwicklungsstadien von Plasmodium yoelii im Nagetier （Ratte） durch ChrysantheMum morifolium

Mannliche Wistar Ratten wurden intravenos mit 700 ×103 Sporozoiten von Plasmodium yoelii infiziert. Den Versuchstieren wurde 2 und 24 Stunden nach der Infektion 700 mg/kg eines Extraktes von Chrysanthemum morifolium in die Bauchhohle gespritzt. 40 Stunden nach der Infektion wurden die Tiere getotet und in Leberdunnschnitten wurde die exoerythrozytare Schizogonie bestimmt. Es wurde gefunden, daβ zwar das Eindringen der Sporozoiten in die Leberzellen nicht gehemmt wurde,jedoch die intrazellulare Reifung zu infektiosen Formen der exoerythrozytare Schizogonie deutlich gehemmt war.

Wirkung des Athylazetatextraktes von Chrysanthemum morifolium auf Plasmodium falciparum In Vitro

Die Wirkung des Athylazetatextraktes von Chrysanthemum morifolium auf P. falciparum wurde in vitro untersucht. Wahrend sich die Parasitamie bei der Kontrollgruppe bis zu 10. 90% entwickelte, wurde bei dem mit Athylazetatextrakt in einer Konzentration von 100 μg/ml Medium am 5. Tag der Kultur eine signifikante Hemmwirkung auf die Parasiten beobachtet. Die Parasitamie blieb bei 1.00 %.

Effect of Spraying Liquorices Root Extract on Vegetative Growth, Chemical Characters and Photosynthetic Pigments of Chrysanthemum （Chrysanthemum morifolium L.） Grown under Salt Stress

This experiment was conducted in Horticulture Department, School of Plant Production, Faculty of Agriculture and Forestry, University of Duhok, Iraq in 2010 on Chrysanthemum plants, in order to mitigate the adverse influence of salinity by the use of liquorice root extract and study its effect on vegetative growth, chlorophyll content in leaves from chlorophyll and assistant pigment, carbohydrate and starch. Subsequently, the trail consisted of three levels of salinity （0, 4 and 8 ds m^-1） and three concentrations of the liquorice root extract 0, 2.5 and 5 g L^-1. Results showed that salinity （4 ds m^-1） rate caused significant increases in certain vegetative characteristics （branches number, plant height, leaves number and leaf area）, photosynthetic pigments （Chl a, Chl b, total chlorophyll, a-carotene, β-carotene, zeaxanthin, astaxanthin）, fresh shoot weight, fresh root weight, shoot dry weight, root dry weight, shoot： root ratio, carbohydrate dry weight percentage and starch percentage as compared to untreated check. However, increasing salinity rate to 8 ds m^-1 resulted in substantial reductions in all above mentioned Waits. Treatment with liquorices root extract, in particular, 5 g L^-1 manifested a potent capability in ameliorating the salinity negative effects in terms of vegetative and photosynthetic pigments aspects.

基于主成分及聚类分析的药用菊花品种产量与品质综合评价

为防止品种混杂,筛选药用或茶用菊花的专用品种,以不同产地来源的8个药用菊花品种16个样品为试材,采用超高效液相色谱法和比色法测定不同产地药用菊花的品质成分含量;并基于主成分分析和聚类分析,对不同产地药用菊花品种产量与品质进行综合评价。结果表明,不同产地药用菊花品种单株花头数变异系数(39.03%)最大,是构成药用菊花单株花头干重的主要因素(r=0.800),以枣庄红心菊单株花头数(496.50)最多、单株花头干重最高;绿原酸、3,5-O-二咖啡酰基奎宁酸和总黄酮等药效成分在不同来源的样本中差异显著(P<0.05),上述三种成分在滁州产滁菊中相对含量较高,分别为3.75%、8.07%、16.08%,这与其品质成分综合评价结果一致,说明滁州滁菊药效较优。13个性状指标的主成分分析提取出药效因子、高产因子、总黄酮因子和花径因子4个主成分,其累计贡献率为84.34%;综合评价得分排在前3位的依次为枣庄引种的红心菊、早小洋菊、滁菊。结合聚类分析结果,判定在原产地,滁州滁菊适于药用;在枣庄引种地,红心菊为优选的高产品种,早小洋菊为综合品质优良的品种。通过建立药用菊花品种产量和品质的综合评价体系,可为我国药茶两用菊花新品种的引种筛选和药食同源利用提供理论依据。

不同直插型基质对3种盆栽菊花生长的影响以及综合评价

为提高盆栽菊花(Chrysanthemum morifolium)扦插繁殖的成活率,提高菊花生产效率,以既有扦插基质又有营养基质的直插型基质代替传统栽培基质,设置了12种不同扦插基质,配合前期筛选的营养基质,扦插培养3种盆栽菊花‘辉煌’‘波尔多红’‘黄芙蓉’。测量其生长、生理以及光合指标,并利用主成分分析和隶属函数对不同直插型基质的盆栽菊花生长效果进行综合评价。结果表明:扦插基质为Z1(河沙)、Z2(珍珠岩)、Z3(蛭石)时,3个盆栽菊花品种的株高、茎粗、冠幅及花期等生长指标更具优势。‘辉煌’‘波尔多红’‘黄芙蓉’在Z2处理中的综合评价指数最高,分别为0.999、0.940、1.000,说明其育苗效果最好,可作为最佳直插型基质,其次为Z1、Z3处理。而扦插基质中,椰糠、稻壳和泥炭体积占比较大时,并不利于盆栽菊花的生长。

怀菊花茎叶中聚炔类化合物对ox-LDL诱导的HUVECs细胞损伤的保护作用

研究怀菊花茎叶中分离得到的聚炔类化合物对氧化低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)损伤的保护作用及初步探讨作用机制。采用ox-LDL(200μg/mL)处理HUVECs细胞建立内皮细胞损伤模型,同时给予聚炔类化合物1~10(100μg/mL)干预;通过CCK-8法检测细胞生存率,筛选出有细胞保护作用的目标化合物;采用目标化合物5和9(12.5、25、50、100μg/mL)干预细胞24 h,CCK-8检测细胞生存率,采用流式细胞术检测细胞凋亡率和活性氧(reactive oxygen species,ROS)生成率,酶联免疫(ELISA)检测细胞上清液中白介素-1β(interleukin-1β,IL-1β)、白介素-6(interleukin-6,IL-6)及肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量;细胞免疫印迹法(In-Cell Western)检测细胞中核转录因子-κB(nuclear transcription factor-κB,NF-κB)2p100/p52与NF-κBp65蛋白的相对表达水平。结果显示:与正常对照组(Con)相比,模型组(ox-LDL)中的细胞生存率明显下降,细胞凋亡率、ROS生成,炎性因子IL-1β、IL-6及TNF-α明显升高(P<0.01);与ox-LDL组相比,聚炔类化合物5和9可以明显提高HUVECs细胞的生存率、降低细胞凋亡率、ROS生成率,炎性因子IL-1β、IL-6及TNF-α的分泌(P<0.05或P<0.01),下调NF-κB2 p100/p52、NF-κB p65蛋白的表达(P<0.05或P<0.01)。综上所述,怀菊花茎叶中聚炔苷类成分可以改善ox-LDL诱导的HUVECs细胞损伤与功能障碍,其作用机制可能与NF-κB信号通路相关。

菊花中216种农药残留检测分析

目的:建立菊花中216种常用农药残留的快速检测方法。方法:样品经乙腈高速匀浆提取后浓缩,采用液相色谱-质谱法和气相色谱-质谱法,以多反应监测模式进行数据采集,采用空白基质匹配法及校准曲线法进行定量检测。结果:菊花中216种农药的线性关系良好,90%以上农药指标r>0.995,各待测农药指标回收率为70%~115%,符合农药多残留的痕量分析要求;78批菊花中检出农药68种。结论:菊花中农药残留检出率较高、检出品种较多,但检出的多为中低毒性农药。建立的方法有针对性地检测了菊花中高风险的农药品种,涵盖了大多数常用农药品种,检测方法快速、准确,适用于菊花中农药多残留的风险监测和日常快速筛查,可为基质类似的中药材农药多残留检测方法研究提供借参考。

大亳菊总黄酮的提取及其颗粒剂制备工艺研究

优选大亳菊总黄酮的提取及其颗粒剂制备工艺。采用正交试验,以总黄酮提取率为判断标准,优选大亳菊总黄酮的提取工艺;以成型率为判断标准,优选大亳菊总黄酮颗粒剂的制备工艺。结果表明,优选的大亳菊总黄酮提取工艺为:液料比为25∶1,乙醇体积分数75%,提取温度85℃,回流时间1.25h;优选的制粒工艺为:可溶性淀粉∶糊精=1∶1,浸膏粉∶辅料=1∶3,干燥温度60℃,干燥时间2h。该工艺简便、稳定,可以用于大亳菊总黄酮的提取及其颗粒剂的制备。

基于谱-效关系筛选菊茎叶总黄酮抗氧化物质基础及其机制探讨

为探讨菊茎叶总黄酮(total flavonoids from stems and leaves of Chrysanthemum morifolium,TFCSL)抗氧化应激的活性成分,阐明其药效物质基础和作用机制。采用HPLC建立不同批次TFCSL指纹图谱;以高浓度葡萄糖诱导人脐静脉内皮细胞建立氧化损伤模型,将丙二醛含量、乳酸脱氢酶含量和超氧化物歧化酶活性作为药效指标;采用灰色关联度和偏最小二乘法分析其谱-效关系确定抗氧化药效物质基础;基于网络药理学结合分子对接探究核心靶点及作用通路。从12批次TFCSL指纹图谱中确定12个共有峰,指认其中9个化学成分;各批次总黄酮样品均可减少细胞凋亡、降低丙二醛及乳酸脱氢酶含量、提高超氧化物歧化酶活性;综合2种数学模型确定峰5(芹菜素-7-O-β-D-葡萄糖苷)、峰6(异绿原酸C)、峰7(香叶木素-7-O-β-D-葡萄糖苷)为抗氧化物质基础;筛选出的3个活性成分作用于抗氧化应激的33个靶点;关键靶点为TNF、CASP3、EDNRA、XDH、PTGS2、MMP2,主要涉及脂质和动脉粥样硬化信号通路、IL-17信号通路、糖尿病并发症AGE-RAGE信号通路、TNF通路、MPKA通路等信号通路;分子对接结果显示活性成分与关键靶点之间均有较好的结合力。表明TFCSL抗氧化应激的物质基础可能为芹菜素-7-O-β-D-葡萄糖苷、异绿原酸C、香叶木素-7-O-β-D-葡萄糖苷,推测通过TNF、CASP3等靶点作用动脉粥样硬化信号通路、IL17信号通路发挥作用,体现菊茎叶多成分、多靶点抗氧化的作用特点。

功能性菊花资源分类及产品开发研究进展

功能性菊花是产业融合性能优异的特色经济作物,市场发展潜力巨大。本文对常见的36种功能性菊花进行了资源分类和特性解析,提出了功能菊花在观赏、茶用、药用以及食用方面的用途,从特色加工产品、景观应用产品及旅游服务产品三个方面探讨了功能性菊花的产品开发策略,以期为功能性菊花的产业化发展提供参考。