Daminozide Alters Anthocyanin Metabolism in Ray Florets of Bronze Chrysanthemum (Chrysanthemum morifolium Ramat.)

Daminozide is a well-known chemical inhibitor of the gibberellic acid biosynthesis pathway regulating the vegetative growth of potted chrysanthemums (Chrysanthemum morifolium Ramat.). However, the precise mechanism underlying daminozide-related floral color loss is unknown. To investigate the latter, in two separate greenhouse experiments, bronze flowering chrysanthemum cultivars ‘Baton Rouge’ and ‘Pelee’ were treated weekly with consecutive (0 or 5,000 mg l^?1) foliar daminozide spray applications at early, intermediate, and late stages during the short-day photoperiod. The ray florets of both cultivars were sampled, and the effect of daminozide application on anthocyanins and their biosynthetic precursors were determined by HPLC. Daminozide applied to ‘Baton Rouge’ plants at early developmental stages was correlated with partial loss of red color, and HPLC analysis determined that this was associated with a 75 % reduction in ray floret anthocyanins. Conversely, a near complete loss of red coloration in daminozide-sprayed ‘Pelee’ relative to control plants was associated with as much as a 98 % decline in anthocyanins, irrespective of the time of application. HPLC analysis determined that daminozide application was associated with a 22–50 % increase in the flavones apigenin 7-O-rutinoside, acacetin 7-O-rutinoside, diosmetin 7-O-rutinoside, and eupatorin, and a 68 % increase in the flavonol quercetin 3-O-glucoside, in ray florets of ‘Pelee’ relative to control plants. There was no relative change in ‘Baton Rouge’ flavone and flavonol levels. The accumulation of bronze C. morifolium flavones and flavonols following foliar daminozide application suggests that red color loss is associated with inhibition of anthocyanidin synthase of ‘Pelee’ ray florets.     

热激锻炼诱导菊花耐热性研究

以菊花(Chrysanthemum morifolium)叶片和花瓣为试验材料,在38℃下对其进行5 h的热激锻炼,再于50℃下分别进行0、0.5、1.0、1.5、3.0 h的高温胁迫,然后对其细胞膜透性、叶绿素、丙二醛(MDA)和蛋白质含量动态变化及5′-核苷酸酶的活性进行了测定.结果表明,热锻炼的菊花叶片电解质渗漏率相对较小,叶片叶绿素含量比对照下降的慢,5′-核苷酸酶活性比对照高,MDA含量也相对较低,而且花瓣中可溶性蛋白质保持了相对较高的含量,说明热锻炼诱导菊花获得了一定的耐热性.     

菊花的化学成分研究

通过硅胶、Sephadex LH-20反复柱层析、纯化,从菊花的乙醇提取物中分离得到14个化合物,利用波谱方法分离鉴定为香叶木素(chrysoeriol,1),木犀草素(luteolin,2),芹菜素(apigenin,3),香叶木素7-O--βD-葡萄糖苷(chrysoeriol 7-O--βD-glucopyranoside,4),木犀草素7-O--βD-葡萄糖苷(luteolin 7-O--βD-glucopyranoside,5),金合欢素7-O--βD-葡萄糖苷(acacetin 7-O--βD-glucopyranoside,6),刺槐苷(acacetin 7-O--βD-glucopyranosyl(6→1)--αL-rhamnopy-ranoside,7),金合欢素7-O-(6″-O-乙酰)--βD-葡萄糖苷(acacetin 7-O-(6″-O-acetyl)--βD-glucopyranoside,8),金合欢素(acacetin,9),山奈酚(kaempferol,10),异泽兰黄素(eupatilin,11),大黄素(emodin,12),大黄酚(chrysophanol,13)和大黄素甲醚(physcion,14)。其中化合物8~14均为首次从菊花中分离得到。     

菊花中一个新的多糖的研究

通过热水提取、离子交换层析和凝胶层析,从菊花(Chrysanthemum morifolium Ramat．)中分离得到一个新的多糖。根据糖组成分析、甲基化分析、高碘酸氧化、部分酸水解和NMR谱图分析,确定其结构如下：     

菊花中一个新的多糖的研究

通过热水提取、离子交换层析和凝胶层析,从菊花(Chrysanthemum morifolium Ramat.)中分离得到一个新的多糖.根据糖组成分析、甲基化分析、高碘酸氧化、部分酸水解和NMR谱图分析,确定其结构如下:[→4)-Galp-(1→4)-Galp-(1→4)-Galp-(1→4)-Galp-(1→6)-Galp-(1]→F3L2Araf-(1→5)-Araf-(1→4)-Glcp-(1→4)-Glcp1→6 F3[→4)-Galp-(1→4)-Galp-(1→4)-Galp-(1→4)-Ga6p-(1→6)-Galp-(1]→n     

Antimutagenic activity of flavonoids from Chrysanthemum morifolium

A methanol extract from the flower heads of Chrysanthemum morifolium showed a suppressive effect on umu gene expression of the SOS response in Salmonella typhimurium TA1535/pSK1002 against the mutagen 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide). The methanol extract was re-extracted with hexane, chloroform, ethyl acetate, butanol, and water. The ethyl acetate fraction showed a suppressive effect. Suppressive compounds in the ethyl acetate fraction were isolated by silica gel column chromatography and identified as the flavonoids acacetin (1), apigenin (2), luteolin (3), and quercetin (4) by EI-MS, IR, and (1)H and 13C NMR spectroscopy. Compounds 1-4 suppressed the furylfuramide-induced SOS response in the umu test. Compounds 1-4 suppressed 60.2, 75.7, 90.0, and 66.6% of the SOS-inducing activity at a concentration of 0.70 micromol/ml. The ID50 (50% inhibitory dose) values of 1-4 were 0.62, 0.55, 0.44, and 0.59 micromol/ml. These compounds had the suppressive effects on umu gene expression of the SOS response against other mutagens, 4-nitroquinolin 1-oxide (4NQO) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), which do not require liver-metabolizing enzymes. These compounds also showed the suppression of SOS-inducing activity against the other mutagens aflatoxin B1 (AfB1) and 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), which require liver-metabolizing enzymes, and UV irradiation. In addition to the antimutagenic activities of these compounds against furylfuramide, Trp-P-1 and activated Trp-P-1 were also assayed by the Ames test using S. typhimurium TA100.     

黄山贡菊的组织培养与快速繁殖

1　植物名称　菊 (Ｃｈｒｙｓａｎｔｈｅｍｕｍｍｏｒｉｆｏｌｉｕｍ)品种黄山贡菊。2　材料类别　茎尖。3　培养条件　 (1 )诱导愈伤组织培养基 :ＭＳ 6 ＢＡ 3ｍｇ·Ｌ- 1 (单位下同 ) ＮＡＡ 0 .1 ;(2 )丛生芽增殖培养基 :ＭＳ 6 ＢＡ 0 .5 ;(3 )生根培养基 :1 /2ＭＳ。上述培养基均加入 0 .8%琼脂和 3 %的蔗糖 ,ｐＨ值为 6 .0 ,在 1 2 1℃高温高压下灭菌 3 0ｍｉｎ。培养温度为 (2 4± 2 )℃ ,光照时间 1 2ｈ·ｄ- 1 ,光照度为 2 0 0 0ｌｘ。4　生长与分化情况4.1　愈伤组织的诱导　从无病虫害、生长健壮的植株上选取茎尖 ,用稀释 …     

Uptake and Distribution of Copper in Chrysanthemum morifolium

The effects of various levels of copper on the uptake and distributionof copper in Chrysanthemum morifolium grown in solution cultureand peat-sand have been examined. Whole plants growing in shortdays were sampled at regular intervals, divided into roots,stem, leaves and lateral shoots, and analysed for copper. Thepartitioning of copper between these tissues showed that a relativelylarge proportion (30–40 per cent) of the total plant copperwas accumulated in the roots of normal plants during the harvestingperiod, compared with approximately 10 per cent in the rootsof copper deficient plants. Whilst the copper content (ug g^–1) of leaves and stemfrom normal plants was negatively correlated with the amountof dry matter produced (P < 0·001), the correspondingcopper deficient tissues showed little variation in copper contentwith increases in tissue dry weight. A more detailed investigationof the copper content of leaves from normal plants showed thatgradients existed within the plant with respect to both leafposition and time of harvest which could be described by a singlecubic surface equation (P < 0·001).     

MicroRNA Expression Profile during Aphid Feeding in Chrysanthemum (Chrysanthemum morifolium)

MicroRNAs (miRNAs) are important regulators of gene expression, affecting many biological processes. As yet, their roles in the response of chrysanthemum to aphid feeding have not been explored. Here, the identity and abundance of miRNAs induced by aphid infestation have been obtained using high-throughput Illumina sequencing platform. Three leaf small RNA libraries were generated, one from plants infested with the aphid Macrosiphoniella sanbourni (library A), one from plants with mock puncture treatment (library M), and the third from untreated control plants (library CK). A total of 7,944,797, 7,605,251 and 9,244,002 clean unique reads, ranging from 18 to 30 nucleotides (nt) in length, were obtained from library CK, A and M, respectively. As a result, 303 conserved miRNAs belonging to 276 miRNAs families and 234 potential novel miRNAs were detected in chrysanthemum leaf, out of which 80, 100 and 79 significantly differentially expressed miRNAs were identified in the comparison of CK-VS-A, CK-VS-M and M-VS-A, respectively. Several of the differentially abundant miRNAs (in particular miR159a, miR160a, miR393a) may be associated with the plant''s response to aphid infestation.     

菊花多糖的研究进展

菊花多糖是菊花中重要的活性成分,日益受到人们的重视。文章介绍了菊花多糖的结构、提取、分离、纯化、测定方法、生物活性等的研究进展,并对其发展前景进行了展望,为深入研究菊花多糖提供参考。     

菊花R病毒杭白菊分离株全基因组序列测定与分析

杭白菊作为著名的中药“浙八味”之一,种植规模和产区不断扩大,但其病毒病的发生也日益严重,对其产量和品质造成严重影响。本研究利用双链RNA（double stranded RNA,dsRNA）和非序列依赖PCR扩增（sequence independent amplification,SIA）等方法,对感病杭白菊病原物进行鉴定,为杭白菊病毒病原的检测构建一套快速和简便的方法。结果表明,感病杭白菊被菊花R病毒（Chrysanthemum virus R,CVR）侵染,将其命名为CVR-TX。通过对其全基因组进行序列扩增与分析,获得其全长基因组为8 872 bp,编码6个ORF,具有Carlavirus属病毒的典型特征。基于全基因组核酸序列以及复制酶、外壳蛋白氨基酸的序列比对发现,CVR-TX与CVR-BJ同源性最高,分别为85.5%、96.0%和96.3%;与Carlavirus属其他病毒同源性分别在48.2%～54.4%、46.9%～55.3%和36.8%～59.5%,因此CVR被确定为一种新的Carlavirus属病毒。系统进化分析表明,基于全长基因组、复制酶（replicase）基因和外壳蛋白（coat protein,CP）基因与CVR-BJ聚为一簇,亲缘性最近。本研究获得了CVR-TX的全长基因组,丰富了CVR的基因组信息,通过生物信息学分析明确其种属关系和区域变化情况,从而为建立CVR可靠灵敏的分子检测手段和有效的防控措施提供理论基础。     

不同品种菊花和贵州产野菊花中木犀草素的含量比较

目的测定并比较不同品种菊花和贵州产野菊花中木犀草素的含量。方法用高效液相色谱法测定木犀草素的含量。色谱柱为ODS柱（4．6mm×250mm,5μm）,流动相为甲醇-水-冰醋酸（体积比45：55：0．4）,检测波长为254nm,流速1．0mL／min。结果7个菊花样品的木犀草素含量,以贵州野菊花的木犀草素含量最高（3．876mg／g）,杭菊的木犀草素含量最低（0．302mg／g）。结论贵州产野菊花中木犀草素的含量均高于其他品种菊花,具有较高的药用价值。     

菊花R病毒杭白菊分离株全基因组序列测定与分析

杭白菊作为著名的中药“浙八味”之一,种植规模和产区不断扩大,但其病毒病的发生也日益严重,对其产量和品质造成严重影响。本研究利用双链RNA（double stranded RNA,dsRNA）和非序列依赖PCR扩增（sequence independent amplification,SIA）等方法,对感病杭白菊病原物进行鉴定,为杭白菊病毒病原的检测构建一套快速和简便的方法。结果表明,感病杭白菊被菊花R病毒（Chrysanthemum virus R,CVR）侵染,将其命名为CVR-TX。通过对其全基因组进行序列扩增与分析,获得其全长基因组为8 872 bp,编码6个ORF,具有Carlavirus属病毒的典型特征。基于全基因组核酸序列以及复制酶、外壳蛋白氨基酸的序列比对发现,CVR-TX与CVR-BJ同源性最高,分别为85.5%、96.0%和96.3%;与Carlavirus属其他病毒同源性分别在48.2%～54.4%、46.9%～55.3%和36.8%～59.5%,因此CVR被确定为一种新的Carlavirus属病毒。系统进化分析表明,基于全长基因组、复制酶（replicase）基因和外壳蛋白（coat protein,CP）基因与CVR-BJ聚为一簇,亲缘性最近。本研究获得了CVR-TX的全长基因组,丰富了CVR的基因组信息,通过生物信息学分析明确其种属关系和区域变化情况,从而为建立CVR可靠灵敏的分子检测手段和有效的防控措施提供理论基础。     

亳菊花中黄酮类化合物的分离鉴定

菊花为菊科植物菊(Chrysanthemum morifolium Ramat.)的干燥头状花序,为了进一步揭示药理活性的物质基础,为该中药的开发利用和质量评价提供依据,安徽亳州亳菊花中的黄酮类化合物进行了系统研究.毫菊经醇提、萃取、硅胶柱层析与纯化得到4个黄酮类化合物.通过理化常数和波谱解析鉴定了其中3个化舍物的结构,分别为:芹菜素-7-甲醚(Apigenin-7-methyl ether)(Ⅰ)、芹菜素(Apigenin)(Ⅱ)和未犀草素(Luteolin)(Ⅲ),另外1个化合物的结构尚在鉴定中.上述3个化合物均为首次从该植物中分到的已知合物.     

Altered expression of CmNRRa changes flowering time of Chrysanthemum morifolium

Flowering time is an important ornamental trait for chrysanthemum (Chrysanthemum morifolium, Dendranthema x grandiflorum) floricultural production. In this study, CmNRRa, an orthologous gene of OsNRRa that regulates root growth in response to nutrient stress in rice, was identified from Chrysanthemum and its role in flowering time was studied. The entire CmNRRa cDNA sequence was determined using a combinatorial PCR approach along with 5′ and 3′ RACE methods. CmNRRa expression levels in various tissues were monitored by real‐time RT‐PCR. CmNRRa was strongly expressed in flower buds and peduncles, suggesting that CmNRRa plays a regulatory role in floral development. To investigate the biological function of CmNRRa in chrysanthemums, overexpression and knockdown of CmNRRa were carried out using transgenic Chrysanthemum plants generated through Agrobacterium‐mediated transformation. CmNRRa expression levels in the transgenic plants were assayed by real‐time RT‐PCR and Northern blot analysis. The transgenic plants showed altered flowering times compared with nontransgenic plants. CmNRRa‐RNAi transgenic plants flowered 40–64 days earlier, while CmNRRa‐overexpressing plants exhibited a delayed flowering phenotype. These results revealed a negative effect of CmNRRa on flowering time modulation. Alteration of CmNRRa expression levels might be an effective means of controlling flowering time in Chrysanthemum. These results possess potential application in molecular breeding of chrysanthemums that production year‐round, and may improve commercial chrysanthemum production in the flower industry.     

Genetic Mapping of Quantitative Trait Loci Underlying Flowering Time in Chrysanthemum (Chrysanthemum morifolium)

Flowering time is an important trait in chrysanthemum, but its genetic basis remains poorly understood. An intra-specific mapping population bred from the cross between the autumn-flowering cultivar ‘Yuhualuoying’ and the summer-flowering ‘Aoyunhanxiao’ was used to determine the number and relative effect of QTL segregating for five measures of flowering time. From flowering time data recorded over two consecutive seasons, 35 additive QTL were detected, each explaining between 5.8% and 22.7% of the overall phenotypic variance. Of these, 13 were detected in both years. Nine genomic regions harboring QTL for at least two of the five traits were identified. Ten pairs of loci epistatically determined the flowering time, but their contribution to the overall phenotypic variance was less than for the additive QTL. The results suggest that flowering time in chrysanthemum is principally governed by main effect QTL but that epistasis also contributes to the genetic architecture of the trait, and the major QTL identified herein are useful in our ongoing efforts to streamline the improvement of chrysanthemum via the use of molecular methodology.     

Rapid mass propagation of Chrysanthemum morifolium by callus derived from stem and leaf explants

A procedure for rapid multiplication of Chrysanthemum morifolium RAMAT cv. Birbal Sahni using leaf callus and stem (nodal/internodal) callus as well as node and apical shoots has been developed. Murashige and Skoog's medium (1962) supplemented with 2mg/1 2,4-D yielded good green calli from both leaf and stem segments within 2 weeks. About 1 cm × 1 cm callus regenerated 2–3 shoots after 3 weeks on MS solid medium supplemented with 0.1 mg/l IAA and 0.2 mg/l BAP. Each of the regenerated shoots when transferred to the same shooting medium without agar yielded about 150 new shoots, which in turn regenerated roots after another week in MS half strength or modified White's media (Rangaswamy, 1961). It has been estimated that about 10¹⁴ plantlets could be produced in a year from one expiant following the proposed protocol.Abbreviations BAP 6-benzylaminopurine - IAA indole-3-acetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - MS Murashige and Skoog's (1962) medium     

Polyamine metabolism,floral initiation and floral development in Chrysanthemum (Chrysanthemum morifolium Ramat.)

In the short-day plant Chrysanthemum (Chrysanthemum morifolium Ramat. variety Pavo) putrescine and spermidine conjugates appeared in the apical bud before the first observable transformation of the meristem into floral structures. These compounds accumulated on floral initiation and well before floral evocation. Spermidine conjugates were predominant during floral initiation whereas free amines did not accumulate to any significant extent. Different associations of amides were observed during floral initiation as compared with the reproductive phase. 3,4-Dimethoxyphenethylamine conjugates (water-insoluble compounds) were the predominant amine conjugates observed during flower development. These compounds decreased drastically after fertilization. In vegetative buds from plants grown in long days polyamine conjugates were very low and appeared as plants aged. We present evidence that ornithine decarboxylase (ODC) regulates putrescine biosynthesis during floral initiation and floral development. When ODC action was blocked by DFMO (-DL-difluoromethylornithine, a specific, irreversible inhibitor of ODC), flowering was inhibited, and free and conjugated polyamines were not detected. This treatment led to a slight enhancement of ADC activity. When putrescine was added, polyamine titers and flowering were restored. A similar treatment with DFMA (-DL difluoromethylarginine, a specific, irreversible inhibitor of ADC) did not affect flowering and the polyamine titers. The results suggest that ODC and polyamine conjugates are involved in regulating floral initiation in Chrysanthemum.Abbreviations ADC arginine decarboxylase - ODC ornithine decarboxylase - DFMA -DL-difluoromethylarginine - DFMO -DL-difluoromethylornithine     

Xanthine oxidase inhibitory effects of the constituents of Chrysanthemum morifolium stems

The flowers of Chrysanthemum morifolium Ramat. (family: Asteraceae) are used in Traditional Chinese Medicine to treat fever, cold and swelling. However the aerial part is considered as agriculture waste and the chemical and pharmacological information is scanty. From the stems of this plant, four new compounds named as morineoliganosides A (1), B (2), C (3) and heterophyllol-1-O-β-d-glucopyranoside A (4) together with 27 known isolates (5–31), including flavonoids and caffeoylquinic acids were isolated. Their structures were elucidated by chemical and spectroscopic methods The inhibitory effects on xanthine oxidase of these compounds have been determined. This research provided a scientific development and utilization of C. morifolium stems.