Development Mode and Countermeasures of Paeonia suffruticosa Andr Industrialization

The research introduced Paeonia suffruticosa industry from market demand, value and adaptability and its characteristics, providing references for Paeonia suffruticosa industrialization from product aspect. Finally, its development models were proposed, as well as countermeasures, providing suggestions and countermeasures for Paeonia suffruticosa industry.

Identification of differentially expressed genes associated with bud dormancy release in tree peony(Paeonia suffruticosa) by suppression subtractive hybridization

A subtractive cDNA library was developed to study genes associated with bud dormancy release in tree peonies. In order to identify genes that are highly expressed in buds released from dormancy, 588 clones were examined by differential screening. Of these, 185 clones were selected to be sequenced. A total of 37 unique sequences were obtained of which only 31 sequences have matches in the NCBI database or the Arabidopsis thaliana protein database. Semi-quantitative RT-PCR was used to confirm further the expression profiles for 12 transcripts identified within the subtractive cDNA library. Gene ontology analyses indicated that many of the different genes identified have unknown or hypothetical functions while it is speculated that other genes play different mo- lecular roles. In our study, genes involved in bud dormancy release were growth-related or stress-responsive, while low-temperature-induced ribosomal proteins may also play a role in bud dormancy release. Our results provide interesting information for further understanding of the molecular mechanism of bud dormancy release in tree peonies.

Regulation of phytohormones on root primordium initiation and adventitious root formation in the etiolated shoots of Paeonia suffruticosa'Yinfen Jinlin'

Tree peony is well known and sought after for its large, colorful flowers. Its propagation is via vegetative methods. Mech- nisms of the adventitious rooting and the regulation of rooting processes are the principles and techniques of plant propagation and improvement. Microstructures and fluctuations of phytohormones in the adventitious rooting were studied with the etiolated soft- wood shoots of Paeonia suffkuticosa ＇Yinfen Jinlin＇. There are no pre-primordia in the shoots of the cultivar. Adventitious roots are produced in five stages： shoot selection, primordium initiation, primordium growth, conducting tissue differentiation and root protru- sion. Primordia initiated in the cortex. The contents of the endogenous hormones, IAA, ABA and GA, were 5.842, 0.873 and 1.043 nmol·g^-1 FW on the bases of shoots, respectively. CTKs which included isopentenyl adenine （iPA）, zeatin riboside （ZR） and dihy- drozeatin riboside （DHZR） were 0.949, 0.695 and 2.034 nmol·g^-1 FW, respectively. DHZR is active among CTKs. The ratio of IAA to GA, CTK and ABA clearly increased at the stage of primordium initiation, while they showed low levels at the stages of primor- dium growth. The ratios were restored at the shoot levels at the stage of root protrusion. IBA provoked primordia initiation in the cortex, the vascular cambium, the pith and even in the callus induced on the base of shoots. ]AA levels in the treated shoots increased gradually to its highest level （three times of control） at the stage of conducting tissue differentiation. The ratios of IAA to GA, CTK and ABA clearly decreased at the stage of primordium initiation. The ratio of IAA to ABA is regulated at 10：1.

Cloning and analysis of cross-intron genomic gene encoding ACO in Paeonia suffruticosa Andr.

The last step of ethylene biosynthesis in higher plants is to convert 1-aminocyclopropane-1-carboxylic acid （ACC） to ethylene by ACC oxidase （ACO）. In our investigation, a cross-introns genomic DNA gene encoding ACC oxidase, PsgACO, was isolated from Paeonia suffruticosa. It revealed that PsgACO （FJ855434） has 1281 bases, containing four exons and three introns, en- coding 312 amino acids. The four exons stretched from 1 to 105, 217 to 434, 592 to 925 and 1000 to 1281 bases. A splicing junction sequence of all the exon-introns conformed to the GT-AG rule in the cross-intron genomic DNA sequence of the ACO gene. PsgACO showed high homology to many characterized ACC oxidases both at the nucleic acid and amino acid levels. As well, twelve amino acid residues were conserved among many ACOs from other species. A phylogenetic tree analysis indicated that the amino acid of ACOs is quite conserved among the different eudicots. The phylogenetic tree showed that both the tree peony and herbaceous peony are quite isolated taxa. Bioinformatic analysis showed that the molecular weight of ACO is 35.29 kD, with a theoretical pI of 5.25. It is a non-secrete, stable hydrophilic protein, located in the cytoplasm.

Outdoor Domestication Cultivation and Survival Mechanism for Tissue Culture Seedlings of Paeonia suffruticosa

In order to reveal the differences between different peony varieties and facilities,thousands of tissue culture seedlings of 21 Itoh peony varieties imported from abroad were used as test materials,and the peony domestication cultivation method used in the past was used as a control. The technical measures and mechanisms for the outdoor domestication of seedlings and the survival of field transplantation were studied. The results indicate that the main factors and measures for effectively protecting the outdoor transplanting of tissue culture seedlings include cultivating sound and strong seedlings,transplanting in proper period of spring,and providing good ventilating places and facilities,removing leaves,keeping buds and releasing dormancy before transplantation,planting depth should meet the requirement of exposing to the root neck,changing the pots according to the size of seedlings,the substrate should be loose and permeable,seriously disinfected and sterilized,scientific management,and prevention and control of diseases and insect pests. In addition,the survival mechanism was also analyzed,and the reasons and countermeasures for the successful application of peony tissue culture in China were found. Finally,the differences in survival rates between different varieties and facilities were summarized.

GC-MS analysis of essential oil of <i>Paeonia suffruticosa</i>Andrews from ZhaoFen and RouFurong flowers in China

The flowers of ZhaoFen and RouFurong may contain essential oils with natural aromatic ingredients. In the present work, the chemical compositions of essential oil of Paeonia suffruticosa Andrews from the flowers of ZhaoFen and RouFurong grown only in China were investigated by GC-MS analysis. The results indicate that there are 27 constituents in ZhaoFen and 29 constituents in RouFurong, which account for 96.04% and 95.90% of the oils of ZhaoFen and RouFurong, respectively. The major components of the essential oils are character-rized by oxygenated terpenols, and their content is, respectively, 85.06% and 83.47%. The essential oil of Paeonia suffruticosa Andrews was reported for the first time on the aerial parts.

Chemical constituents and theiranti-infective activity of Paeonia suffruticosa

Objective:Research the chemical constituents of Paeonia suffruticosa that are responsible for its anti-infective properties.Methods:Several column chromatographic methods were used to purify the chemical constituents from P.suffruticosa,including medium pressure liquid chromatography,Sephadex LH-20,and normal silica gel.A disc diffusion method was used to screen for antibacterial activity,and their anti-virulence activity was assessed on the type III secretion system(T3SS)of Salmonella pathogenicity island 1(SPI-1)in Salmonella enterica serovar Typhimurium UK-1 g 8956 by SDS-PAGE and western blots.Results:Twenty-one compounds were identified.Compounds 7,8 and 17 showed moderate activity against S.aureus ATCC25923,compounds 8,9 and 10 showed weak activities against B.subtilis ACCC11060.Meanwhile,phenols(14-18)and flavonoids(20 and 21)inhibited T3SS protein secretion of S.typhimurium without affecting bacterial growth.Furthermore,a strong inhibitory effect was observed for 17 and 20 on SPI-1 mediated invasion of HeLa cells.Additionally,no toxicity was observed for these compounds.Conclusion:P.suffruticosa has anti-infective properties due in part to the fact that phenols and flavonoids can block the secretion of T3SS-associated protein effectors.

Paeonia suffruticosa Andrews的界定,兼论栽培牡丹的分类鉴定问题

栽培牡丹一直称为PaeoniasuffruticosaAndrews。在把作丹皮用的栽培牡丹 (凤丹 )描述为新种P .ostii后 ,观赏的栽培牡丹仍然被统称为P .suffruticosa。但在P .suffruticosa复合体内 ,P .rockii,P .jis hanensis,P .qiui和P .suffruticosassp .yinpingmudan等野生类型相继被描述并得到承认。于是 ,有人提出了栽培牡丹多元发生的观点 ,同时也出现了P .suffruticosa“是牡丹组数种植物复合体”的说法。本文在详细论述P .suffruticosa复合体分类历史的基础上 ,根据我们自己对野生类型和栽培类型的考察研究 ,认为观赏栽培牡丹的形成是多元的 ,并且包含几个物种 ;P .suffruticosa有明确的概念 ,它是观赏栽培牡丹的组成部分 ,而不代表栽培牡丹的全部 ,更不是数种植物的复合体。我们认为应根据形态特征正确鉴定观赏栽培牡丹 ,不应把它们一概鉴定为P .suffruticosa。通过文献和标本引证 ,文中界定了P .suf fruticosa的范围。此外 ,把P .yinpingmudan (D .Y .Hong,K .Y .Pan&Z .W .Xie)B .A .Shen作为P .suffruticosassp.yinpingmudanD .Y .Hong,K .Y .Pan&Z .W .Xie的异名 ,并且认定P .yinpingmudan (D .Y .Hong ,K .Y .Pan&Z .W .Xie)B .A .Shenssp .henanensis(D .Y .Hong,K .Y .Pan&Z .W .Xie)B .A .Shen是不?

药用植物凤丹(Paeonia suffruticosa)根际土壤细菌群落16S rRNA基因的ARDRA分析

凤丹(Paeonia suffruticosa Andr.)为芍药科多年生植物,是一种重要的传统中药资源。在其生长发育的周期中与土壤微生物尤其是根际土壤微生物有密切的关系。通过构建16S rRNA基因克隆文库及文库的限制性片段长度多态性分析(ARDRA),对中国药用植物凤丹5大主要分布区域的根际土壤细菌群落多样性进行了研究。采用限制性内切酶HinfI和Csp6I对克隆文库中随机挑选的1000个白色克隆子进行了酶切分型,根据酶切图谱的不同,将其分为324个OTUs,并对38个优势OTUs进行了测序和系统发育分析。16S rRNA基因序列分析结果表明,凤丹根际土壤细菌种群主要包括:变形菌门(包括alpha、beta、gamma、detla亚门)、酸杆菌门、放线菌门、拟杆菌门及厚壁菌门等11类细菌,此外还包含了3个未归类的细菌。变形菌门和酸杆菌门为文库中的主要菌群,分别占克隆总数的47.34%和14.36%,其中Pseudomonas sp.、Burkholderia sp.和Arthrobacter sp.为优势菌属。研究结果表明,我国药用植物凤丹5大主要分布区域的根际土壤细菌种群不仅具有丰富的多样性,还存在丰富的潜在新菌种。

牡丹(Paeonia suffruticosa)导管的形态多样性

利用生物显微镜对牡丹一年生枝条的次生木质部离析材料进行了观察,旨在了解牡丹(Paeonia suffruticosa)的导管形态特征,为其生理生态及系统发育的合理解释及应用提供理论参考。结果表明:牡丹次生木质部具有螺纹、梯纹和孔纹三种类型的导管,穿孔板具有孔状穿孔板和梯状穿孔板两种类型,导管端壁的穿孔及尾尖形态多样,孔膜退化比较彻底。不存在明显的导管二态现象,平均长度为1100μm,平均宽度为200μm。牡丹导管形态特征的多样性揭示了其较为复杂生理生态适应机制和相对较低的演化地位。

芍药(PaeoniaL.)生物学特性研究进展

综述了近年来国内外芍药的研究进展概况,系统地介绍了芍药的起源、分布与分类,栽培与繁殖,组织培养,生殖生物学等方面的研究现状及发展方向。

牡丹花青素苷合成关键基因PsDFR启动子活性分析

[目的]分析牡丹花青素苷合成关键基因PsDFR启动子的顺式作用元件及活性,为进一步研究PsDFR启动子的功能及其在牡丹花色形成中的调控机制奠定基础。[方法]以‘黑花魁’牡丹花瓣中提取的基因组DNA为模板,通过染色体步移法克隆PsDFR的启动子序列。利用生物信息在线软件预测分析启动子序列的顺式作用元件。构建5个不同长度缺失启动子与GUS基因融合的表达载体,并瞬时转化烟草叶片,通过GUS组织化学染色和酶活性检测分析缺失启动子的活性,及其对脱落酸(ABA)、茉莉酸甲酯(MeJA)、光照等不同胁迫处理的响应。[结果]克隆得到PsDFR上游1687 bp的启动子序列。生物信息学分析发现PsDFR启动子含有多个光信号、激素响应、逆境响应及组织特异表达等顺式作用元件,这暗示PsDFR的表达可能受光信号、激素和胁迫等多种信号的共同调节。GUS组织化学染色和酶活性检测表明,随启动子片段的缩短,启动子活性逐渐下降,-1623至-916区段对于启动子活性具有重要作用。MeJA和黑暗处理对各启动子片段活性均有显著抑制作用,恢复光照可促使启动子活性明显回升,而参与ABA响应的核心调控区域位于-443至-76 bp。[结论]PsDFR启动子含有多个光信号、激素响应、逆境响应及组织特异表达等顺式作用元件,其活性受光的正调控以及MeJA的负调控;-1623至-916 bp间的区域对于启动子活性具有重要作用,-443至-76 bp是响应ABA处理的核心区域。该研究为进一步揭示PsDFR应答环境信号参与牡丹花色形成的分子调控机制提供参考。

牡丹基因组AFLP银染反应体系的建立和优化

采用改进的SDS方法，从牡丹成熟叶片中提取基因组DNA，所得DNA样品的A260／A280值在1．7～1．8之间，琼脂糖凝胶电泳主带清晰、DNA纯度高、较少降解、不舍酶反应抑制剂，可被MseⅠ和PstⅠ两种限制性内切酶完全酶切．通过酶切连接、预扩增、选择性扩增、银染等试验过程，成功建立了牡丹品种AFLP银染反应体系。得到了清晰的指纹图谱，对扩增结果进行聚类分析，得到了23个牡丹品种的聚类分析图。

HPLC法测定丹皮中丹皮酚的含量

牡丹皮为毛茛科牡丹Paeonia suffruticosa Andr.的根皮,常用作清热、凉血药,丹皮酚是其主要成分之一,具有降压、止血等多种药理作用。丹皮酚含量测定方法有直接蒸馏-UV法、HPLC法等。

牡丹不同连栽年限对春季土壤动物群落的影响

【目的】了解牡丹(Paeonia suffruticosa)不同连栽年限对春季土壤动物群落结构的影响。【方法】以毗邻荒地(CK)为对照,大型土壤动物采用手捡法,中小型土壤动物采用干湿漏斗分离法,研究了牡丹连栽1 a(F1)、2 a(F2)、3 a(F3)和4 a(F4)后土壤动物群落特征。【结果】试验共捕获土壤动物190只,隶属于2门7纲15目31科,优势类群为蚰蜒目和蜈蚣目,与CK相比,牡丹种植整体上增加了土壤动物类群数,并随连栽年限的增加而呈现出抛物线的变化趋势,牡丹连栽3 a后(F3),土壤动物类群数和平均密度达到最高;同时,除第3年外,牡丹种植后降低了其余种植年的土壤动物密度。随着连栽年限的增加,土壤动物丰富度指数呈递增趋势,多样性指数呈抛物线的变化趋势,优势度指数呈现逐渐减少的趋势,均匀度指数值波动较小。捕食性与杂食性土壤动物密度随连栽年限的增加均呈先递增后递减的趋势。【结论】牡丹种植在短期内有利于土壤动物群落发展,但在超过一定年限后,土壤动物群落会受到一定抑制。

部分牡丹品种遗传多样性的AFLP分析

目的研究牡丹遗传多样性为合理利用牡丹种质资源、培育观赏价值高的牡丹新品种提供依据。方法利用荧光标记AFLP技术,采用8对M+3和P+3引物组合对30份牡丹栽培品种进行了总基因组DNA水平上的多态性检测。结果检测结果共获得1123条可统计的条带,其中965条呈多态性,多态性带百分率达86%。揭示了牡丹栽培种质丰富的遗传多样性。8组引物在30个品种中检测到数目不等的品种特异带型,这些品种的特异带型对供试牡丹品种具有一定的鉴别价值。结论8对引物能将30个牡丹品种完全区分开。聚类分析结果表明多数来源地相同的牡丹种质表现出较为密切的亲缘关系。

河南部分牡丹品种遗传多样性的AFLP分析

利用AFLP技术对30个牡丹品种的遗传多样性进行了研究。用3对引物组合进行选择性扩增,共产生151个位点,其中96个为多态性位点,占63.5%。其中2个受试品种产生了特异性位点。聚类分析结果显示,遗传距离0.25将30个牡丹品种划分为5个聚类组,与传统分类结果基本一致。

HPLC法测定牡丹籽油中游离型和结合型α-亚麻酸、亚油酸及油酸的含量

采用高效液相色谱(HPLC)法测定牡丹籽油中游离型、结合型α-亚麻酸、亚油酸及油酸的含量。检测条件为Agilent C18高效色谱柱(3.0 mm×50 mm,2.7μm),流动相乙腈-0.1%磷酸水溶液(体积比85∶15),流速1 mL/min,检测波长203 nm,柱温35℃,进样量20μL。结果表明,该方法精密度、重复性、稳定性良好。α-亚麻酸在41.4~207μg/mL范围内,其峰面积与质量浓度呈良好的线性关系;亚油酸在20.2~101μg/mL范围内,其峰面积与质量浓度呈良好的线性关系;油酸在14.9~74.6μg/mL范围内,其峰面积与质量浓度呈良好的线性关系。牡丹籽油样品中,游离型α-亚麻酸、亚油酸和油酸的平均含量分别为4.47、1.89 mg/mL和0.92 mg/mL,结合型α-亚麻酸、亚油酸和油酸的平均含量分别为437、237 mg/mL和156 mg/mL。建立的方法具有分离效率高、选择性好和条件温和的优点。

基于牡丹涝害胁迫的转录组分析及SSR引物开发

涝害胁迫是限制牡丹种植、生长和高产的最主要的非生物胁迫之一。为了阐明牡丹在涝害胁迫下的作用机制,研究进行了转录组学分析,并开发了SSR标记。利用高通量测序对6个由涝害处理的幼苗和对照的mRNA构建的cDNA文库进行测序。通过组装总共获得73925个基因,创建了牡丹的初始参考转录组数据库。其中780个被鉴定为涝害早期反应基因,其中155个基因上调,625个基因下调。功能分析表明,参与转录因子信号调节、DNA复制、核糖体和嘧啶代谢的基因表达的改变可能在牡丹对涝害胁迫的反应中发挥重要作用。基于这些高通量转录组测序数据,挖掘出5204个SSR标记。在这些标记中,二核苷酸重复占58.13%,三核苷酸重复占27.11%,四核苷酸重复占7.24%。在牡丹耐涝性状靶基因相关的功能注释基因中,发现了110对SSR标记引物,筛选出45对引物,其中12对能扩增出清晰的条带,多态性SSR标记引物占引物总数的26.67%。研究结果不仅有助于阐明牡丹的涝害反应机制,同时也为耐涝牡丹品种的分子标记辅助选择育种提供了宝贵的基因组资源和科学依据。

基于叶绿体psbA-trnH序列和trnL-F序列的牡丹野生种间关系

对牡丹组全部9个野生种15个居群及凤丹的叶绿体psb A-trn H和trn L-F序列进行测序,并采用邻位相连法分别构建了psb A-trn H序列和trn L-F序列的系统发育树。结果显示:1)两段序列的基因树均支持牡丹组划分为两个亚组的分类方法,与前人的研究结果一致。2)psb A-trn H序列基因树表明大花黄牡丹与黄牡丹的亲缘关系最近,这与大花黄牡丹曾被确定为黄牡丹的一个变种的历史一致。3)革质花盘亚组内,基于psb A-trn H和trn L-F序列的研究结果大体一致,分歧主要在四川牡丹的地位问题。4)psb A-trn H序列和trn L-F序列基因树分别表明杨山牡丹与凤丹具有最近或较近的亲缘关系。