Semisynthetic Derivatives of Sissotrin Isolated from Trifolium baccarinii Chiov. (Fabaceae) and Evaluation of Their Antibacterial Activities

Starting from sissotrin (1), a natural isoflavonoid isolated from Trifolium baccarinii (Fabaceae), one new semisynthetic derivative, 6-nitrobiochanin A (1b) and two known derivatives, 8-nitrobiochanin A (1a) and 2",3",4",6"-tetraacetylsissotrin (1c) have been obtained after performing nitration and acetylation reactions. Their structures were assigned after interpretation of their spectrometric (HR-ESI-MS) and spectroscopic (NMR 1D and 2D) data and by comparison with those reported in the literature. The substrate as well as the semisynthetic derivatives were evaluated for their antibacterial activities against six strains. The results reveal that they are inactive or weakly active on the strains tested with the exception of 8-nitrobiochanin A (1a) which showed moderate activity (MIC = 62.5 μg∙mL−1) on Staphylococcus aureus ATCC 43300.

白三叶(Trifolium repens)入侵对城市草坪生态系统土壤动物的影响

土壤动物是土壤生态系统中十分活跃的生物类群之一,也是城市草坪生态系统的重要组成部分。由于外来植物白三叶入侵,草坪土壤动物可能受到直接或间接的影响,进而改变城市草坪生态系统功能及过程。研究白三叶入侵对城市草坪生态系统土壤动物的影响,可为进一步了解外来植物入侵机制及城市草坪的建植养护提供理论依据。采用野外调查的试验方法对4种不同入侵程度下城市草坪的土壤动物群落特征和土壤理化性质进行了研究。结果如下:4种不同入侵程度的草坪样地共捕获土壤动物30099只,隶属于19目43科,其中小杆科、线蚓科和跳虫科为优势类群;土壤动物个体数量和类群数量整体表现为中度入侵>轻度入侵>对照>重度入侵,符合中度干扰假说;土壤动物个体数量和类群数量的季节动态主要表现为夏秋较高,冬春较低。CCA分析显示,土壤理化因子中铵态N对土壤动物影响相对较小,可能与白三叶改善了草坪土壤的供N水平并使其不成为土壤动物分布的限制因子有关;同时,土壤理化因子对土壤动物优势类群影响较小,而主要影响常见和稀有类群。总体而言,一定程度的白三叶入侵增加了土壤动物群落的多样性,但当达到重度入侵时,白三叶形成单优群落,减少了植物多样性,进而使土壤动物可获得的生活资源减少,土壤动物群落多样性呈下降趋势;白三叶入侵后通过改变土壤理化性质,影响到土壤动物常见和稀有类群,并最终改变草坪土壤动物的群落结构。

土壤水分梯度对白三叶(Trifolium repens)光合作用和根系分布的影响

通过对两个品种白三叶Trifolium repenscv.Haifa（海发）和Trifolium repenscv.Rivendel（瑞文德）盆栽试验,模拟3种不同的土壤水分状况（无水分胁迫：保持植株良好的水分供应;轻度胁迫：表层0-20 cm土壤处于干旱状态;重度胁迫：表层0-20 cm土壤处于极干旱状态,20-40 cm土壤处于干旱状态）对白三叶光合作用和根系生长的影响。结果表明,当植株未遭受水分胁迫时,两个品种白三叶的光合作用和根系生长状况没有明显差异;当表层0-20 cm处于干旱状态时,‘海发’在处理后期的净光合速率和水分利用效率升高,根系生长量增大,表现出促进作用,‘瑞文德’受到的影响不显著;当表层0-20 cm处于极干旱、20-40cm处于干旱状态时,‘海发’在处理前期受到轻微影响,随后恢复正常状态,‘瑞文德’则受到较严重的影响。随着干旱程度的加深和时间的延长,白三叶的根冠比逐渐增大。与‘瑞文德’相比,在相同时期相同胁迫程度下,‘海发’的根冠比没有显著差异,但深根数量大大超过‘瑞文德’,因而,‘海发’的耐旱能力强于‘瑞文德’。

迷果芹(Sphallerocarpus gracilis)和红三叶(Trifolium pratense)的核型分析

对迷果芹 ( Sphallerocarpus gracilis( Bess.) K- Pol.)和红三叶 ( Trifolium pratenseL.)进行了染色体计数及核型分析。迷果芹的染色体数目为 2 n=2 0 ,核型公式为 K( 2 n) =2 x=2 0 =1 4m+ 4 sm+ 2 st( SAT) ;核型类型为 2 A,为较对称核型 ,该种植物的染色体数目及核型均为首次报道。红三叶的染色体数目有 2 n=1 4、1 6、2 8、32等类型 ,本研究首次报道了 2 n=1 4的核型公式为 K( 2 n) =2 x=1 4=2 M+ 1 2 m,核型类型为 1 B。

Antibacterial activity of leaves extracts of Trifolium alexandrinum Linn,against pathogenic bacteria causing tropical diseases

Objective:To investigate antibacterial potential of Trifolium alexandrinum(T.alexandrinum) Linn,against seven gram positive and eleven gram negative hospital isolated human pathogenic bacterial strains responsible for many tropical diseases.Methods:Non-polar and polar extracts of the leaves of T.alexandrinum i.e.,hexane,dichloromethane(DCM),ethyl acetate(EtOAc), methanol(MeOH) and aqueous(AQ) extracts at five different concentrations(1,2,5,10 and 15 mg/ mL) were prepared to evaluate their antibacterial value.NCCL standards were strictly followed to perform antimicrobial disc susceptibility test using disc diffusion method.Results:Polar extracts demonstrated significant antibacterial activity against tested pathogens.EtOAc and MeOH extracts showed maximum antibacterial activity with higher inhibition zone and were found effective against seventeen of the tested pathogens.While AQ plant extract inhibited the growth of sixteen of the test strains.EtOAc and MeOH plant extracts inhibited the growth of all seven gram positive and ten of the gram negative bacterial strains.Conclusions:The present study strongly confirms the effectiveness of crude leaves extracts against tested human pathogenic bacterial strains causing several tropical diseases.Since Egyptian clover is used as a fodder plant,it could be helpful in controlling various infectious diseases associated with cattle as well.

白三叶(Trifolium repens L)无性系植物分株间的生理整合作用

研究了白三叶无性系植物种群，在相互连接的分株之间生理上相互依存的程度。结果显示子代在生理上取决于母株资源的转移，这一依赖性随时间而下降。相互连接的分株，一部分被遮荫至１０％，而照光分株光合速率明显增加。光被限制期间，分株将重新整合，遮荫分株无性系中由其它分株提供同化物。在子代匍匐茎数目和长度上有一个权衡。当资源受限制时，产生极少较长的子代匍匐茎，结果提高了子代能够占据更有利条件的可能性。在分株之间，生理整合作用降低了子代建立所面临的危险，使同源分株在资源的可利用能力上整合到一个局部的异质环境，增加同源分株间的竞争能力，缓冲所面对的局部微生境的选择。

Phenolic Profiles and Polyphenol Oxidase (PPO) Gene Expression of Red Clover (Trifolium pratense) Selected for Decreased Postharvest Browning

Red clover (Trifolium pratense L.) is a legume forage abundant in phenolic compounds. It tends to brown when cut for hay, due to oxidation of phenolic compounds catalyzed by polyphenol oxidase (PPO), and subsequent binding to proteins. Selecting for a greener hay may provide information about the relationship of browning, PPO, and phenolics to each other. The red clover Kenland cultivar was selected over eight breeding cycles for decreased browning after being cut and dried 48 h in the field. Expression of PPO1 and PPO3, in Kenland and three of the eight cycles, was compared by real-time quantitative PCR. Phenolic compounds in Kenland and Cycle 8, collected 0, 24, and 48 h after cutting, were quantified by high-performance liquid chromatography (HPLC). Visual browning scores decreased 12% between Kenland and Cycle 8 (P = 0.02). PPO1 and PPO3 gene expression were not affected by selection. Clovamide decreased 26% in Cycle 8 relative to Kenland (P = 0.016). Sissotrin decreased 10% in Cycle 8 (P = 0.043). Neither total formononetin nor total biochanin A was affected by selection (P = 0.63 and 0.45, respectively). These results suggest that when selecting clover for decreased postharvest browning, a decrease occurs in a phenolic compound that can bind protein independently of PPO. However, PPO1 and PPO3 gene expression, and the major red clover isoflavones, are minimally affected.

红三叶(Trifolium pratense)的染色体核型分析

本文对红三叶（Ｔｒｉｆｏｌｉｕｍ ｐｒａｔｅｎｓｅ）的染色体数目及核型进行了初步研究，结果表明，其染色体数目为２ｎ＝１４，核型公式为Ｋ_（２ｎ）＝２ｘ＝１４＝２Ｍ＝１２ｍ，即７对染色体中，１对为正中着丝粒染色体，６对为中央着丝粒染色体。

白车轴草(Trifolium repens)植株抗病性和生长与植物病史的关系(英文)

从白车轴草 (Trifoliumrepens)自然种群中采集无白车轴草单孢锈菌病史的无性系 (clones) 17个 ,有白车轴草单孢锈菌病史的无性系 14个 ,分别作为抗病型和感受型植物实验材料 ;采集白车轴草单孢锈菌 (Uromycestrifolii repentis)菌系 (strains) 10个 ,作为病菌实验材料 .分别设置并进行了两个温室实验、一个田间盆栽实验和一个原生长地移栽实验 ,实验处理上分对照、单菌系接种和 10个菌系接种等 3种 .实验结果表明 ,无论是用单菌系接种还是 10个菌系接种 ,植株发病的概率和程度均与其抗病性有关 ,抗病型植株 (无病史 )发病的概率和程度显著低于感受型 (有病史 )植株 .在相同处理的实验中 (无论是田间实验还是温室实验 ) ,无病史植株和有病史植株的生长无显著差异 ;不同处理田间实验植株的生长有显著差异 ,病情愈重 ,生长愈差 .无病史植株的抗病性明显强于有病史植株 .但是 ,原生长地的移栽实验结果表明 ,在无病原菌存在的情况下 ,有病史植株的 (叶 )生长显著好于无病史植株 .可以认为 ,研究生物个体对环境因子反应性差异的实验应当在自然条件下和自然梯度范围内进行 .

Phenotypic and Genetic Characterization of Trifolium Rhizobia Isolated from Temperate and Subtropical Regions of China

The clovers that comprise Trifolium genus are naturalized in the subtropical and tropical zones in China. They are valuable bioresources as important green manures and pasture grass, which contribute biologically fixed dinitrogen （N2） and provide nutrition to farming systems. However, there are very few effective strains available for inoculant production and there is little information available about symbiotic rhizobia in Chinese legume clover root nodules. In this study, 139 root nodule bacteria were isolated from two clover species （Trifolium repens and Trifolium pretense） growing in the subtropical and temperate regions of China, 16S rRNA gene sequence analysis, BOX-PCR, whole cell protein SDS-PAGE, and nodulation tests were performed to characterize these strains. The results showed that phenotypic and genetic diversities among 139 isolates were large, with 83 protein patterns and 66 BOX- AIR profiles, respectively. The rhizobial strains were first divided into two large phenotypic protein groups. The sequencing strains representing the two groups were related to Rhizobium leguminosarum USDA2370T and R. sophorae CCBAU03386T and had 99.6%-100% similarities. The phylogeny specificity of the rhizobia from clover was elucidated, while showed a large variation in the fingerprints of the phenotypes and genotypes and genetic diversity was high （revealed by Shannon diversity index, H＇）. The rhizobial isolates from subtropical regions, such as Anhui Province, Yunnan Province and Hubei Province, had higher diversities than those from temperate areas, such as Hebei Province and Shanxi Province, which could be used to identify rhizobial strains from clover and screen efficient inoculum strains. A number of diverse rhizobial strains had been identified and a pool of currently available clover rhizobial strains were increased. This would ultimately increase the likelihood of identifying more efficient strains suited for developing a successful inoculation strategy for the production of white clover.

A new hydroxybenzoic acid glycosides from the plant of Trifolium lupinaster

A new 5-O-(β-D-glucopyranosyl)-6 -O-benzoyl-2,5-dihydroxybenzoic acid was isolated from the ethyl acetate extract from the whole plants of Trifolium lupinaster L.Its structure was determined and deduced on the basis of their spectral data.

Construction of Clonal Growth Models in Trifolium repens

The suitability of models for describing the clonal growth of Trifolium repens population was discussed. The results showed that deterministic models were inadequate for describing its clonal growth, but the diffusion models and the randomwalk models suited for the clonal growth characteristics of the population. And it was found that random-walk models were better than diffusion models for describing a population in an environment with rich natural resources, and the latter was better in a poor environment.

Changes in Florets’Vertical Direction within Inflorescence Affects Pollinator Behavior,and Fitness in Trifolium repens

interactions,many flowers are known to display their attractive qualities,such as scent emission,flower rewards and floral vertical direction,in a rhythmic fashion.However,less is known about how plants regulate the relationship between these flower traits to adapt to pollinator visiting behavior and increase reproduction success.Here we investigated the adaptive significance of the flower bending from erect to downward in Trifolium repens.We observed the flowering dynamic characteristics(changes of vertical direction of florets,flowering number,pollen grain numbers,pollen viability and stigma receptivity over time after blossom)and the factors affecting the rate of flower bending in T.repens.Then we altered the vertical direction of florets in inflorescence of different types(upright and downward),and compared the pollinator behaviors and female reproductive success.Our results showed that florets opened sequentially in inflorescence,and then bend downwards slowly after flowering.The bending speed of florets was mainly influenced by pollination,and bending angle increased with the prolongation of flowering time,while the pollen germination rate,stigma receptivity and nectar secretion has a rhythm of“lowhigh-low”during the whole period with the time going.The visiting frequency of all the four species of pollinators on upward flowers was significantly higher than that of downward flowers,and they especially prefer to visit flowers with a bending angle of 30°–60°,when the flowers was exactly of the highest flower rewards(nectar secretion and number of pollen grains),stigma receptivity and pollen germination rate.The seed set ratio and fruit set ratio of upward flowers were significantly higher than downward flowers,but significantly lower than unmanipulated flowers.Our results indicated that the T.repens could increase female and male fitness by accurate pollination.The most suitable flower angle saves pollinators’visiting energy and enables them to obtain the highest nectar rewards.This coordination between plants and pollinators maximizes the interests of them,which is a crucial factor in initiating specialized plant-pollinator relationships.

基于SSR分子标记的13个白三叶(Trifolium repens L.)品种指纹图谱构建

白三叶(Trifolium repens L.)是一种多年生冷季型豆科牧草,因其具有产量高、品质优良等特点,已成为我国西南地区重要的优质牧草。为分析国外引进的白三叶新品种的遗传多样性,本研究利用简单重复序列(Simple sequence repeats,SSR)分子标记对13个白三叶品种进行品种鉴定及DNA指纹图谱的构建,以期利用此方法直接、有效的鉴定和区分白三叶品种。结果表明:20对SSR引物共扩增出115条不同的条带,其中多态性条带104条(90.4%);同时所有引物的多态信息含量(Polymorphism information content,PIC)范围为0.198~0.455,平均值为0.333;AMOVA分析表明,49%的遗传变异发生在品种间,51%发生在品种内部;聚类分析和主成分分析的结果表明,13个品种的亲缘关系中,‘超级海法’和‘海法’的亲缘关系最密切,‘米莉格’和‘克朗德’的遗传距离较远;利用6种具有代表性的引物gtrs1164,gtrs573,gtrs541,gtrs171,gtrs536和gtrs173构建的DNA指纹图谱具有较强的识别能力,可将13个白三叶品种有效的鉴定区别开来。综上所述,本试验所选用的6对引物具有良好的多态性,可用于白三叶品种的指纹图谱构建。

Volatile Constituents of Leaves of Trifolium alexandrinum

This study presents the volatile constituents from leaves of Trifolium alexandrinum from Pakistan.The dried leaves were subjected to hydrodistillation for extraction and extracted volatile oil was subsequently characterized by gas chromatography-mass spectrometry technique.A total of 22 compounds were identified where the most dominant constituents were:phytol(46.00%),palmitic acid(9.26%),phytol acetate(6.44%),and linolenic acid(3.47%).Most of these are well-known constituents to exhibit antibacterial,antioxidant,antitumor,and anti-inflammatory activities.

适生植物白三叶草(Trifolium repens)对陇东地区不同浓度油污土壤场地生态修复的响应

为了明确白三叶草(Trifolium repens)生物量和土壤环境指标对不同浓度油污土壤场地生态修复的响应方式,采用常规方法测定了不同油污浓度胁迫下其根际土壤总石油烃(TPH)降解率、生物量指标、根际微生物数量和酶活性的变化情况。结果显示:(1)随着油污浓度增加,根际TPH降解率依次为75. 49±2. 14%、75. 57±2. 52%,55. 02±2. 06%、33. 14±2. 22%和13. 34±1. 53%;(2) 70000mg/kg-1时其出芽时间延迟至15. 61±1. 65d,而出芽率、植物体干重和株高则分别下降了38. 11%、40. 44%和51. 18%;(3)当油污浓度≤50000mg/kg时土壤脱氢酶、多酚氧化酶以及土壤烷烃和芳烃降解菌数量均与油污浓度均呈正相关关系(P <0. 01),但在70000mg/kg时均受到显著抑制(P <0. 05);(4) 50000和70000mg/kg时植物生物量指标完全受到抑制,其根际土壤微生物特性和土壤酶活性高低是决定重度污染TPH降解的主要环境因素。

三叶草(Trifolium repens)用于土壤镉污染的修复潜力

通过温室盆栽试验法,研究了土壤镉(Cd)污染下三叶草对土壤Cd的富集特征及土壤Cd的净化能力等,以期为三叶草对土壤Cd的生态修复提供依据。结果表明:三叶草具有较好的Cd耐受性,总生物量增加值介于2.0~3.7 g。随着土壤Cd处理浓度增加,三叶草根、茎、叶中Cd含量分别高达178.6、101.3、130.9 mg·kg-1,富集系数(BF)分别介于9.7~17.9、3.0~10.1、3.1~13.1,富集能力大小总体表现为根>叶>茎,转运系数(TF)值均≥1(除T2处理),表明三叶草有较强的Cd富集能力,且能较好地将Cd转运至地上部位;三叶草根、茎、叶对Cd的吸收量均随着土壤Cd处理浓度的升高而递增,地上部位(茎、叶)吸Cd百分率(占总吸Cd量)最高达87.2%,对土壤Cd净化率最高达6.2%。因此得出三叶草具有作为Cd超积累植物的较好潜力,可以进行污染区域美化改造和Cd污染修复。

Resource Sharing by the Different Clonal White Clover Plant(Trifolium repens L.)

Resource sharing in the different parts of the clonal white clover plant (Trifolium repens L.) was investigated in this study. The results showed that the integration of water and nitrogen metabolism occured among parental, young and old branches of a plant when water passed through its stolons. Under the sufficient supply of light, water and nutrients from soil the newly developed branches showed similar capacity in taking in sufficient water and nitrogen to that of old branches. The development of the new branches did not affect the growth of the old and parental branches attached. The results also indicated that energy values in roots was significantly related with plant age, however, the energy values were various in the different parts of a plant.

Characterization of the copper-containing amine oxidase from Trifolium pratense seedlings

We studied the kinetic characterizations of the Trifolium pratense seedlings copper-containing amine oxidase(TPAO) by using various amine-containing substrates.The catalyzing rate for all of amine-containing substrates can be ordered as diamines > polyamines > aromatic monoamines,and it shows an apparent trend in each category of substrates such as the longer the carbon chain,the lower the V_(max) is,so does the V_(max)/K_m values but is opposite for the K_m value of TPAO.The distinct differences between the kinetic parameters for different amine-containing substrates indicated that the rate-determining step of the catalytic reaction strongly depends on the substrate's chemical structure.It is concluded that both pH and ionic strength can affect the catalytic activity of TPAO via influencing the coulomb interaction-mediated enzyme-substrate docking processes,which can be attributed to the potential of charged groups from both substrates and the activity sites of TPAO by the regulation of ionic strength.

配施有机肥对白三叶草生长及香蕉园土壤生态的改良作用

为探明香蕉园间作白三叶草(Trifolium repens L.)模式下配施纯羊粪和生物有机肥对白三叶草的生物量和营养元素含量及香蕉园土壤理化性质的影响,采用间作白三叶草+常规管理(CK)、间作白三叶草+施用纯羊粪、间作白三叶草+施用生物有机肥3个处理做对比分析。结果表明,间作条件下配施纯羊粪,白三叶草地上部和地下部的生物量分别显著增加17%和10%,地上部和地下部的N、P、K含量分别显著增加21%、33%、17%和13%、15%、53%,土壤有机质、pH、全氮和速效钾含量分别显著增加17%、3%、22%和3%;间作条件下配施生物有机肥,白三叶草地上部和地下部的生物量分别显著增加19%和25%,地上部和地下部的N、P、K含量分别显著增加26%、12%、5%和44%、50%、22%,土壤有机质、pH、全氮和速效钾含量分别显著增加42%、4%、33%和4%。由此可见,在香蕉园间作白三叶草模式下配施纯羊粪或生物有机肥均能显著提高白三叶草的生物量和N、P、K元素含量,同时还能改善香蕉园土壤生态,且配施生物有机肥优于配施纯羊粪。