A Genetic Transformation System for Rosa multiflora Thunb. var. cathayensis Rehd. et Wils through Callus Induction

An efficient genetic transformation system is a preparation for Rosa multi-flora Thunb. var. cathayensis Rehd. et Wils to diversify its flower color through ge-netic engineering. We firstly optimized the explants and culture conditions on callus induction, hormone concentrations and dark period of culture time on bud differentia-tions in particular, with sterilized seedlings to establish the regeneration system of R. multiflora. It showed that callus induction frequency reached 100% after the ex-plants being cultured in dark for 21 d when MS was chosen to be the initial culture medium. The bud differentiation rate was 48% after cal i being cultured under dark for 8 d on MS medium supplemented with TDZ （1.5 mg/L） and NAA （0.05 mg/L）. The cal i was used as the explants that were infected with Agrobacterium tumefa-ciens harboring a DFR-RNAi construct. The transformation rate reached as high as 50%. The establishment of a highly efficient rose gene transformation system out-lined in this report is prerequisite for genetic improvement in rose flower colors.

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.

根癌农杆菌胁迫下多花蔷薇(Rosamultiflora‘Inermis4’)的基因表达分析

多花蔷薇无刺4号(Rosamultiflora‘Inermis4’)对根癌病有较强抗性。以根癌农杆菌(Agrobacteriumtumefaciens)接种多花蔷薇无刺4号为目的材料,刺伤处理为对照,采用抑制差减杂交(suppressionsubtractivehybridization,SSH)方法,构建了富集多花蔷薇根癌病抗性相关基因的差减cDNA文库。差异筛选差减cDNA文库后,随机选取增强表达的92个克隆进行测序,去除冗余的cDNA和病菌的基因片段,共得到53个单一序列。利用GenBank数据库进行核酸和蛋白质同源性比较,获得的cDNA片段功能涉及:细胞分裂、生长(22个克隆),逆境防御、信号转导及激素调节(11个克隆)以及初生与次生代谢(10个克隆)。另有10个基因片段功能未知,可能为新的EST。根据测序及比对分析结果,对多花蔷薇无刺4号抗根癌病的分子机理进行了探讨。

部分蔷薇(Rosa multiflora)属植物的RAPD分析

蔷薇作为园林观赏植物,在中国城市绿化中应用较为广泛,但目前应用的蔷薇品种较为混杂,尚无系统研究。本研究采用RAPD技术对部分蔷薇属植物进行亲缘关系的分析,旨在进一步明确蔷薇种、品种,为引种育种以及材料选择提供参考。对蔷薇属植物的2个种、1个变种、20个品种进行了RAPD分析,16个引物对供试蔷薇材料PCR扩增获得175条谱带,其中多态性条带151条(86.3%)表现出丰富的遗传多态性。且不同花色系蔷薇属植物间的遗传多态性不同,白色系75.16%,粉红色系73.46%,黄色系37.50%。利用UPGMA法构建树状聚类图,蔷薇属植物的2个种、1个变种、20个品种在阈值为0.667处被分划为两大类群,这两大类群间的遗传距离较近,黄蔷薇与黄刺枚单独被划分为一个类群,与上述两类群间的遗传距离较远。聚类结果显示出了蔷薇的演化趋势:白色、单瓣是蔷薇较为原始的一个性状,花径大、重瓣性高是现代蔷薇栽培品种的性状,而部分粉红色半重瓣蔷薇则是从野生种到现代栽培品种的中间过渡类型。

In vivoand in vitroanti-arthritic efficacy of a herbal formula consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos

OBJECTIVE Rheumatoid arthritis(RA)is the most common inflammatory autoimmune disease,affecting around 1% of the world population.Toll-like receptor 4(TLR4)signalling has been found to be involved in the pathogenesis of RA.It is a potential therapeutic target for RA treatment.A herbal formula(RL)consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos has traditionally been used in treating various inflammatory disorders.In this study,we would evaluate the anti-arthritic effect of RL on collagen-induced arthritis(CIA)in rats and investigate the involvement of TLR4 signaling in the mode of action of RL in vivo and in vitro.METHODS In vivo anti-arthritic efficacy was evaluated using CIA rats induced by bovine typeⅡ collagen.The treatment groups were treated with various concentrations of RL or positive control indomethacin for 35 d.Clinical signs(hind paw volume and arthritis severity scores),changes in serum inflammatory mediators,histological and radiographic changes of joints were investigated.Spleens and peritoneal macrophages were used to determine the effects of RL on innate and adaptive immune responses in CIA rats.The involvement of TLR4 signalling pathways in the anti-arthritic effect of RL was examined in cartilage tissue of CIA rats,murine RAW264.7macrophages and human THP-1 monocytic cells.RESULTS The severity of arthritis in the CIA rats was significantly attenuated by RL.Histological score and radiographic score were efficiently improved by RL.RL could also dose-dependently inhibit pro-inflammatory cytokines in serum of CIA rats.RL significantly inhibited the production of various pro-inflammatory mediators,the expression and/or activity of the components of TLR4 signalling pathways in animal tissue and cell lines.CONCLUSION RL possesses anti-arthritic effect on collagen-induced arthritis in rats.The therapeutic effect of RL may be related to its inhibition on pro-inflammatory cytokines in serum.The inhibition of the TLR4/TAK1/NF-κB and TLR4/TAK1/MAPK pathways participate in the anti-arthritic effects of RL.This provides a pharmacological justification for the use of RL in the control of various arthritic diseases.Further investigation should be done to develop RL into a modern anti-arthritic agent.

Gas Chromatography-Based Fingerprinting and Chemical Pattern Recognition of Rosa multiflora Volatile Oil

In this paper, gas chromatography-mass spectrometry （GS-MS） was used to build the standard fingerprint of volatile oil from Rosa multiflora Thunb. from 12 different habitats. Fourteen components in the volatile oil were identified as the indicator components ofR. multiflora, of which one was selected as the standard. The GC analysis conditions used for fingerprinting afford a very good separating effect. The similarity of the 12 volatile oils from R. multiflora Thunb. was more than 0.84, and the precision, stability and repeatability of the fingerprints were quite good. It could be concluded that the fingerprints can be used as the standard and as a quality control method for medicinal materials from R. multiflora Thunb..

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.

野蔷薇根总黄酮对类风湿关节炎模型大鼠血管损伤的改善作用及机制

目的研究野蔷薇根总黄酮对类风湿关节炎(RA)模型大鼠血管损伤的改善作用及其潜在机制。方法将雌性Wistar大鼠随机分为正常对照组、模型组、阿司匹林组(阳性对照,30mg/kg)和野蔷薇根总黄酮低、高剂量组(4.15、8.30g/kg,以生药量计),每组10只。除正常对照组外,其余各组大鼠采用胶原诱导+高脂饮食喂养建立RA模型。造模14 d后,各组大鼠灌胃相应药液/0.5%羧甲基纤维素钠溶液,每天1次,连续36d。定期评估大鼠的全身评分、关节炎指数和足爪肿胀数;末次给药后,检测大鼠血清中白细胞介素6(IL-6)、细胞间黏附分子1(ICAM-1)和血管细胞黏附分子1(VCAM-1)水平,观察大鼠胸主动脉血管组织病理学形态变化,检测胸主动脉血管组织中Toll样受体4(TLR4)蛋白表达水平和Janus激酶2(JAK2)、信号转导及转录激活因子3(STAT3)蛋白磷酸化水平。结果与正常对照组比较,模型组大鼠血清中IL-6、ICAM-1、VCAM-1水平和胸主动脉血管组织中TLR4蛋白表达水平及JAK2、STAT3蛋白磷酸化水平均显著升高(P<0.01),胸主动脉血管组织可见动脉粥样斑块(粥瘤)、胆固醇结晶、淋巴细胞浸润及少量未破裂的泡沫细胞聚集。与模型组比较,在第28天时,野蔷薇根总黄酮各剂量组大鼠的全身评分(低剂量组除外)、关节炎指数和足爪肿胀数均显著降低(P<0.05或P<0.01);在第49天时,野蔷薇根总黄酮低剂量组大鼠的全身评分、关节炎指数和足爪肿胀数均显著降低(P<0.05或P<0.01);野蔷薇根总黄酮各剂量组大鼠血清和血管组织中其余定量指标均显著逆转(P<0.05或P<0.01),血管组织病理损伤明显减轻。结论野蔷薇根总黄酮可改善RA模型大鼠的血管损伤,其作用可能与降低血管组织中TLR4蛋白表达,抑制IL-6/JAK2/STAT3信号通路活性有关。

不同品种和产地玫瑰精油成分的主成分与聚类分析

通过检索文献方式收集玫瑰精油成分数据,参考玫瑰精油标准共筛选出37种国内外玫瑰精油作为研究对象。采用主成分分析(PCA)和聚类分析(CA)方法对其成分进行分析,并建立评价模型。PCA结果表明,国内外前4种主成分的累计贡献率分别达到90.765%和75.269%,且可以反映大部分精油成分信息。综合得分排名最前为伊朗地区的突厥蔷薇和山东地区的紫枝玫瑰,其成分含量均匀,品质较好。CA将31种国外玫瑰精油集聚为5大类,6种国内玫瑰精油集聚为3大类,集聚为一类的产地接近,其成分含量具有相似性。

基于顶空固相微萃取-气相色谱-质谱法和相对气味活性值分析不同发酵时间玫瑰花茶香气的差异

目的探究不同发酵时间处理对玫瑰花茶香气及感官品质的影响。方法本实验在相同环境条件下,以玫瑰花瓣为材料,采取萎凋、揉捻、发酵、干燥4道制作工艺,分发酵0 h(CK)、发酵2 h(Treat-2 h)、发酵4 h(Treat-4 h)、发酵6 h(Treat-6 h)4个处理,采取顶空固相微萃取-气相色谱-质谱法(headspace solid phase microextraction-gas chromatography-mass spectrometry,HS-SPME-GC-MS)鉴定玫瑰花挥发性成分,筛选其关键呈香物质。结果共检测出938种16类挥发性代谢物。Treat-2 h、Treat-4 h、Treat-6 h与CK的挥发性代谢物存在显著差异物质554种,相对CK分别差异显著代谢物390种、416种、430种,同时对照4种处理挥发性物质的相对气味活性值(relative odor activity value,r OAV),结合感官评审结果,得出Treat-4 h为玫瑰花茶的最好处理,且发现关键呈香(玫瑰花香气)的物质为壬醇、异戊酸苯乙酯、苯乙醛、2-苯乙醇、香叶醇、大马士酮、苯甲醇、异丁酸香叶酯、(E)-6,10-二甲基-5,9-十一碳烯-2-酮、壬酸乙酯。结论玫瑰花瓣能制作成质量优异的玫瑰花茶,Treat-4 h的玫瑰花茶品质最佳。以上结果阐明了玫瑰发酵过程中香气成分变化,为实现玫瑰花产业资源化利用提供了参考依据。

多花蔷薇假珠芽诱导、体细胞胚发生及植株高效再生

以多花蔷薇‘无刺3号’成熟种子为外植体,探讨了假珠芽诱导、体细胞胚发生及植株高效再生的方法。结果表明,种子的子叶在添加2,4-D0.5-2mg·L^-1的1/2MS培养基上暗培养30d后所产生的愈伤组织,接种到添加TDZ5-20mg·L^-1的1/2MS培养基上光培养20d产生初级假珠芽。假珠芽在添加TDZ10mg·L^-1和GA30.1mg·L^-1的诱导培养基上暗培养20d后,在含麦芽糖的无植物生长调节剂的培养基上光培养产生少量次级假珠芽和胚性愈伤组织。假珠芽保存在诱导培养基上。胚性愈伤组织可发育成大量正常体细胞胚,继而再生正常植物。假珠芽可通过上述方法不断诱导体细胞胚和假珠芽,通过这种方法假珠芽已经保存了2年。

微波消解-火焰原子吸收法测定新疆野蔷薇果中金属元素

采用HNO3消解体系对野蔷薇果进行程序升温微波消解制样,应用火焰原子吸收法测定野蔷薇果中的Ca、K、Mn、Zn、Mg、Fe、Cu、Na、Cr、Ni、Sr共11种金属元素的含量。对样品前处理条件进行筛选,确定样品适合的微波消解体系并进行消化结果的准确度和精密度实验。该方法的加标回收率为96.50%～104.50%,相对标准偏差(RSD)≤3.21%。结果表明,新疆野蔷薇果中富含Ca、Mg、K、Na、Mn、Fe等对人体有益的金属元素,而Cr、Ni等金属元素的含量较低。

红刺玫愈伤组织诱导再生体系的建立

红刺玫（Rosa multiflora Thunb.var.cathayensis）为蔷薇科蔷薇属野蔷薇的一个变种,半常绿灌木,花色粉红。红刺玫以花期长、耐受高温和高湿气候等特性成为园林绿化的新宠[1]。但红刺玫的单瓣花和花色单一的缺陷限制了其广泛的应用,改良红刺玫花色引起园艺工作者的极大兴趣。

野蔷薇根总黄酮的提取及纯化工艺

目的:优化野蔷薇根总黄酮提取及纯化工艺。方法:L9(34)正交试验回流提取确定最适提取剂乙醇浓度、料液比、提取时间组合;从静态吸附-解吸、动态吸附-解吸两方面筛选最佳树脂型号,单因素考察最佳纯化条件。结果:UV 508 nm检测波长下,最佳提取条件为50%乙醇溶剂,料液比1∶10,进行90℃、90 min回流提取;14种大孔树脂中,HPD-400型大孔树脂对野蔷薇根总黄酮的吸附-解吸能力最好,树脂纯化最佳上样浓度为浸膏母液稀释1.5倍;上样量35 m L,流速3 s/滴;除杂用水量40 m L,流速1 s/滴;50%乙醇32 m L洗脱,流速2 s/滴。结论:最优提取纯化条件下,总黄酮回收率达50%以上。

月季和野蔷薇的试管嫁接研究

【目的】研究月季和野蔷薇的试管嫁接技术,为月季繁殖的有效途径提供参考。【方法】以野蔷薇带芽茎段为外植体,通过比较不同灭菌方法、不同激素种类和浓度的培养基对组培快繁的影响,筛选出适宜的灭菌方法以及培养基,从而建立野蔷薇的无菌快繁体系,并以不同品种月季无菌苗的茎尖为接穗,野蔷薇为砧木进行远缘嫁接。【结果】野蔷薇的外植体灭菌宜采用75%酒精溶液拭擦茎段表面后浸泡10 s,再用0.1%的升汞溶液浸泡10 min,继代增殖培养基为MS+6-BA 3.0 mg/L+NAA 0.10 mg/L,生根培养基为WPM+NAA 0.05 mg/L;保留砧木叶片有利于提高嫁接成活率和生根率;以继代增殖培养25 d的月季茎尖为接穗,有利于远缘嫁接的成活。移栽以绯扇-野蔷薇的成活率最高,其次为"卡罗拉-野蔷薇"、"金凤凰-野蔷薇"、"红双喜-野蔷薇"、"龙沙宝石-野蔷薇"、"遗产-野蔷薇"。【结论】不同月季品种与野蔷薇嫁接的成活率因品种而异,砧木保留叶片与否以及接穗的继代培养时间与嫁接成活率有着密切的关系。

野蔷薇根醇提物对大鼠动脉粥样硬化的影响

目的:观察YT对大鼠AS的影响。方法:50只SD大鼠按随机数字表法分为空白组、模型组、YT低、高剂量组、AL组,灌胃给药90 d,每2周称摄食量和体质量,调整灌胃量;空白组喂标准饲料,其余组高脂饮食联合注射维生素D3并加免疫损伤造成AS;末次给药1h后,取主动脉进行病理组织学观察,比色法检测血清中SOD、MDA水平,化学发光法检测血清中CRP水平,ELISA法检测血清中sCD40L、ADNP、IL-18、IL-16水平,放射免疫分析法检测血清中IL-6、TNF-α水平。结果:YT低、高剂量组明显改善AS大鼠主动脉壁增厚、脂质沉积,减少泡沫细胞形成及淋巴细胞浸润;YT低剂量组体质量增加幅度、MDA水平显著降低,摄食量显著增加,YT高剂量组IL-18水平显著降低。结论:YT可通过调节脂质过氧化及炎症反应改善AS。

多花蔷薇总RNA提取方法

根据总RNA完整性、纯度和得率筛选出适合多花蔷薇幼嫩根、叶总RNA的提取方法.结果表明,以CTAB/酸酚法提取的扦插苗根系总RNA、以LiCl-尿素法提取的扦插苗根、叶总RNA以及采用RNeasy Plant Mini Kit试剂盒的改进方法提取的组培苗嫩叶总RNA电泳有清晰明亮的28S、18S条带,无降解;其A260/A280值为1.73～2.04,表明总RNA质量好.RT-PCR结果进一步证实所提取的总RNA能够用于分子生物学的各种下游实验.RNA得率分别为:根系和组培苗嫩叶120-140μg/g(fw),扦插苗嫩叶190-230μg/g(fw).CTAB/酸酚法提取的嫩叶总RNA、SDS/酸酚法提取的根、叶总RNA有多糖污染,且有明显降解.Total RNA isolation system(Z5111,Promega)试剂盒不适合提取多花蔷薇各组织总RNA.

玫瑰花产量灰色分析、Kriging插值及选择指数研究

【目的】探讨玫瑰(Rosa rugosa Thunb.)高产品种的选育理论与技术。【方法】对20个玫瑰品种的15个数量性状与单株产花量进行灰色系统分析、Kriging插值模拟、综合选择指数分析。【结果】单株花数、分枝数、花蕾宽和单花鲜重等4个性状与单株产花量的灰色关联度较大(大于0.5)。根据单株产花量的Kriging插值模拟结果,单株产花量随单株花数、分枝数的增加而增加,任何单一因素的变化均不能保证主要目标性状的提高。【结论】提高玫瑰单株产花量的选育方法应该采用多性状综合选择法,选用单株花数、分枝数、花蕾宽和单花鲜重等4个性状,以关联度为权重建立了单株产花量综合选择指数模型:I=0.3187x1-318.6164x2+670.1262x4+6.3474x8,指数遗传力=0.8014,综合育种值选择进展ΔH=245.8811,该研究结果对今后玫瑰高产花量品种的选育具有指导意义。

微波辅助提取玫瑰花红色素工艺的优化

[目的]优化微提取玫瑰花(Rosa rugosa Thunb)红色素的工艺。[方法]以云南高原食用玫瑰为原料,在单因素试验的基础上,通过正交试验优化微波提取工艺。[结果]微波提取玫瑰花色素的最佳工艺为:微波功率为560 W,提取时间为35 s,料液比为1∶5,提取溶剂为40%乙醇。在最佳工艺条件下,玫瑰花红色素提取率达到81.6%,提取收率达到3.6%。[结论]微波提取技术可较好地提取玫瑰红色素。

玫瑰花发育过程中芳香成分及含量的变化

【目的】研究玫瑰花发育过程中芳香成分及其含量的变化。【方法】采用顶空固相微萃取(HS-SPME)和气相色谱-质谱联用(GC-MS)技术分析‘唐紫’玫瑰花发育过程中芳香成分及其含量的变化。【结果】花蕾期共检出芳香成分53种,萜烯类化合物是主要香气成分,含量较多的芳香成分有γ-依兰油烯、α-雪松烯、α-蒎烯、反式-α-香柠檬烯、顺式-乙酸-3-己烯酯、长叶马鞭草烯酮;初开期共检测出65种芳香成分,醇类、酯类和萜烯类化合物是主要香气成分,含量较多的有β-香茅醇、乙酸香茅酯、苯乙醇、香叶醇、乙酸正己酯、橙花醇、乙酸苯乙酯、顺式-乙酸-3-己烯酯;半开期共检测出芳香成分62种,醇类、酯类和萜烯类化合物是主要香气成分,含量较多的有β-香茅醇、香叶醇、苯乙醇、乙酸香茅酯、橙花醇、乙酸正己酯、乙酸苯乙酯和α-月桂烯;盛开期共检测出芳香成分65种,主要为醇类、酯类和萜烯类化合物,含量较多的有β-香茅醇、乙酸香茅酯、苯乙醇、乙酸香叶酯、香叶醇、乙酸苯乙酯、橙花醇、乙酸正己酯和α-月桂烯;盛开末期共检测出芳香成分58种,主要为醇类、酯类和萜烯类化合物,含量较多的有β-香茅醇、苯乙醇、乙酸苯乙酯、乙酸香茅酯、α-月桂烯、α-蒎烯、柠檬烯和乙酸香叶酯;伴随花的不断发育,醇类化合物含量迅速增加,半开期含量最高;酯类化合物呈先升高后降低的趋势,盛开期含量最高;萜烯类化合物在花蕾期含量最高,初开期迅速下降,至盛开末期又有所上升;醛类化合物的含量呈先迅速升高后迅速降低的趋势,盛开期含量最高。【结论】玫瑰花发育的不同时期,芳香成分及含量差异明显。鲜花的主要香气成分在初开期基本形成,大部分芳香成分在半开期至盛开期含量最高。因此,生产中提取玫瑰精油时,应选择开放程度在半开期至盛开期的鲜花进行采摘。