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.

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。

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。

Response of Chlorophyll Fluorescence Characteristic of Trifolium repens L. to Water Stress

[Objective] The aim was to study response of chlorophyll fluorescence parameters in Trifolium repens L.leaves to water stress from perspective of physiology.[Method] With T.repens cultivar ＂Haifa＂ as tested material,three soil water content levels were set to culture plants,including 75% （no stress,CK）,50% （moderate stress,LD） and 25% （severe stress,HD）,the effects of water stress on chlorophyll fluorescence parameters were determined.[Result] The results showed that the chlorophyll fluorescence parameters changed a little when the relative soil moisture content was 75% and 50%,while the chlorophyll fluorescence parameters such as conversion efficiency of primary light energy （Fv/Fm） of PS Ⅱ,the photochemical quenching coefficient （qP） and the quantum yield of PS Ⅱ electron transport （φPS Ⅱ） decreased respectively when the relative soil moisture content was 25%.[Conclusion] When T.repens in severe drought conditions,the physiological functions were damaged,showing symptoms of drought injury.

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。

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.

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.

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.

氧化石墨烯对白三叶根系生长、生理和解剖结构的影响

为了探究氧化石墨烯(GO)对植物根系特征的影响,以GO和豆科植物白三叶(Trifolium repens L.)为实验材料,通过将不同质量分数的GO(0.2%、0.4%和0.6%)添加到土壤中,进行为期100 d的植物培养,分析了白三叶根系生长、逆境生理、营养吸收及其解剖结构的响应特征.结果显示:随着GO质量分数的增加,白三叶的根系生长和根系活力显著下降,0.6%GO处理组的根长、直径、体积、干重和根系活力较对照组分别降低了31.2%、35.1%、62.5%、94.1%和64.6%.GO处理后根系的抗氧化酶活性显著增加,矿质营养元素含量随着GO质量分数的升高而降低.根系输导组织的解剖结构参数,包括周皮厚度与细胞层数、导管直径与数量、维管柱宽度、木栓层厚度均随GO质量分数的增加逐渐减小.由此可知,GO会诱导白三叶根系氧化胁迫,激活抗氧化酶系统,抑制根系对矿质营养的吸收,破坏根系输导组织结构,最终抑制了根系生长.

HPLC测定红车轴草中大豆苷元、染料木素、芒柄花素和鹰嘴豆芽素A的含量

目的:建立测定红车轴草中大豆苷元、染料木素、芒柄花素和鹰嘴豆芽素A含量的高效液相色谱方法。方法:KromasilC18色谱柱(250mm×4.6mm,5μm),流动相:乙腈0.1H3PO4(40∶60),流速:1.0mL·min-1,检测波长:254nm,柱温:25℃。结果:大豆苷元、染料木素、芒柄花素和鹰嘴豆芽素A的线性范围分别为0.0662～0.6620μg,0.0978～0.9780μg,0.0866～0.8660μg和0.0992～0.9920μg。该方法回收率大豆苷元为98.9(RSD=0.46),染料木素为98.8(RSD=1.21),芒柄花素为98.8(RSD=0.71),鹰嘴豆芽素A为97.5(RSD=1.33)。结论:该法简便、快速、准确。

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。

HPLC法测定红车轴草提取物中有效成分的含量

建立了一种测定红车轴草提取物中大豆素、染料木素、刺芒柄花素含量的高效液相色谱分析方法。Polaris C18色谱柱（150mm×4．6mm，5μm），流动相为乙腈一0．1％H3PO4（40：60），流速1．0mL·min，检测波长260nm，柱温25℃，进样量20μL。结果表明，大豆素、染料木素、刺芒柄花素的线性范围分别为0．40～2．00μg、0．10～0．96μg、0．10～0．96μg，其回收率分别为98．42％（RSD=1．32％）、98．15％（RSD=1．96％）、100．17％（RSD=0．32％）。该法简便、快速、准确。

红车轴草提取物中总鹰嘴豆芽素类成分HPLC测定方法的研究

Two HPLC methods were established to determine the total biochanins in Red Clover extracts.One was taken Ononin,Sissotrin,Formononetin and Biochanin A as external standard.The other was taken Biochanin A as extrernal standard only.The results of two methods were primary similar.

转LeNCED1基因对白三叶ABA含量及水分利用效率的影响

【目的】研究LeNCED1基因对白三叶水分利用效率的影响,为白三叶的节水抗旱提供一定的理论指导。【方法】分别以7株含转SAG13-LeNCED1基因白三叶和非转基因白三叶的老叶和嫩叶为材料,研究其ABA的含量、光合特性、叶绿素含量及水分利用效率。【结果】转SAG13-LeNCED1基因显著提高了白三叶老叶的ABA含量(P<0.05),却显著降低了白三叶老叶的叶绿素b、胞间CO2浓度、蒸腾速率和气孔导度(P<0.05)。转LeNCED1基因显著降低了白三叶嫩叶叶绿素a、叶绿素b、净光合速率和蒸腾速率(P<0.05),却显著提高了白三叶嫩叶的胞间CO2浓度和气孔导度(P<0.05)。【结论】转入LeNCED1基因能显著提高白三叶的长期水分利用效率(P<0.05)。

Development, Reproduction and Body Size Variation of Aphis glycines Matsumura Fed on Different Plants

Soybean aphid, Aphis glycines Matsumura is one of the most important pests in soybean. Life cycle of A. glycines is characterized as heteroecious and holocyclic. The primary hosts of A. glycines are Rhamnus spp. and the secondary hosts include cultivated soybean, Glycine max(L.) Merr, and wild soybean, Glycine soja Sieb.& Zucc. In this study, A. glycines were fed on Trifolium repens L. and Metaplexis japonica(Thunb.) Makino for three generations and their development, reproduction and body sizes were studied. These data were compared to the control fed on the known hosts, G. max and G. soja. These newly deposited offspring by the 3 rd generation were transferred back onto G. max and these differences in their development, reproduction and body sizes were also studied. It showed that A. glycines all could survive, develop and reproduce well, when they were fed on T. repens and M. japonica for three generations, but there were significant differences in their nymph stage, adult longevity and fecundity, life table parameters and body sizes. When these offspring were transferred back onto G. max which were newly deposited by the 3 rd generation aphids fed on T. repens and M. japonica, they could survive, develop and reproduce well, but there were significant differences in their nymph stage, adult longevity, intrinsic rate of increase and body sizes. It provided important information for studies on host adaptability of A. glycines on T. repens and M. japonica and to make clear the potential that this aphid involved into host biotypes on these plants.

Genetic Diversity of Clover by SRAP

In the present study,genetic diversity and genetic relationship of 11materials including red clover ( Trifolium pretense L.),white clover ( T.repen L.) and alsike clover ( T.hybridum L.) and leaf-type variants of white clover were investigated by SRAP (sequence related amplified polymorphism).Forty SRAP primers were screened,and 792 reliable bands were amplified,among which 426 (53.8%) were polymorphic.The number of polymorphic bands per pair of primers ranged from 3 to 38 with an average of 10.6.SRAP cluster analysis showed that the similarity coefficients between white clover materials were from 0.465 to 0.997 with an average of 0.812,indicating that there was certain genetic diversity among white clover.Specific bands appeared in white clover variants,indicating that there were certain differences in DNA sequence between normal while clover plants and their leaf-type variants.The similarity coefficients between red clover materials were from 0.457 to 0.827 with an average of 0.597,indicating that the different red clover germplasms had genetic diversity at the DNA molecular level.The results also showed that the genetic relationship of the alsike clover with the red clover was closer than that with the white clover.This study shows that SRAP technique can be effectively used for the analysis on interspecific and intraspecific relationship,germplasm resource identification and genetic diversity of clover.

Genetic structure and diversity of Ukrainian red clover cultivars revealed by microsatellite markers

Polymorphism of microsatellite loci of Ukrainian red clover cultivars has been studied. 87 microsatellite alleles, which occurred in different combinations, were identified. The number of alleles ranged from 7 to 10. Microsatellite allele distribution showed that 15 alleles were common for all the red clover cultivars (17.2%). The red clover cultivars were represented by homozygous and heterozygous genotypes. The observed and expected heterozygosity ranged from 0.067 to 0.269 and from 0.225 to 0.807, respectively. An analysis of molecular variance revealed that the largest proportion of variation (68.5%) resided at the intrapopulation level. Differentiation of the Ukrainian cultivars was moderately expressed (FST = 0.07).

逆境胁迫对红三叶影响的研究进展

红三叶(Trifolium pratense L.)是豆科车轴草属中的一种,是富含粗蛋白、粗脂肪和黄酮类化合物等的优良多年生牧草和药用植物,在生态建设中也发挥着重要作用。红三叶性喜温暖湿润气候,耐湿性能良好,但耐旱性能较差、抗逆性较弱和生态幅狭窄等限制了在我国大多数地区的推广利用。文章综述了国内外近年来对红三叶重金属胁迫、干旱胁迫、水淹胁迫、生物胁迫及低温胁迫的研究进展,概述了红三叶响应逆境胁迫的生理和分子机理,旨在为今后红三叶新品种选育和在我国草牧业发展中的推广利用提供背景资料和理论参考。

野火球叶片色素和多酚类物质含量与叶色参数的关系

为探明影响野火球(Trifolium lupinaster L.)叶色的关键参数及其叶色差异与叶片色素等相关物质含量的内在联系,本研究采集了翠绿色叶、绿色叶、深绿色叶类型野火球的成熟叶,同时对每类型的叶色特征、叶片色素(叶绿素a,叶绿素b,类胡萝卜素)、多酚类物质含量等进行测定,采用相关分析和主成分分析以探明影响野火球叶色的关键参数。结果表明:翠绿色叶类型的叶片中可溶性糖和花色素苷含量最高;绿色叶类型P n和G s最高,叶绿素总浓度、过氧化物酶活性和单宁含量均最多;深绿色叶类型叶片a^(*),h^(*)和s^(*)值较大。相关分析和主成分分析发现,a^(*),h^(*)和s^(*)与叶绿素含量呈显著正相关;b^(*),C^(*),H值与可溶性糖、花色素苷含量呈显著正相关;h^(*)和s^(*)及色素含量的因子负荷较高。野火球叶色存在差异是因为叶绿素和花色素苷含量不同,导致叶片h^(*)和s^(*)值不同,继而野火球呈现出不同的叶色。

不同处理方式对野火球种子硬实及萌发的影响

采用砂纸打磨+变温、砂纸打磨+恒温两个双因素试验及酸、碱两个单因素试验共四种方式对野火球种子进行处理,筛选破除野火球种子硬实的最佳温度条件以及最佳方法。结果表明:(1)砂纸打磨+恒温、砂纸打磨+变温处理的硬实率显著降低,砂纸打磨+恒温处理以恒温25.0℃处理的萌发及硬实破除效果最佳,砂纸打磨+变温处理以10.4℃/26.6℃为打破野火球种子硬实、促进萌发的最佳变温条件。(2)98%H_(2)SO_(4)处理8 min时野火球种子发芽率略高于对照但无显著差异,但活力指数显著高于对照(P<0.05),硬实率降低,种子受损程度小。(3)野火球种子在0.5 mol/L NaOH处理下的发芽率和活力指数与对照相比均显著降低(P<0.05),对硬实破除无显著效果,死种子数增加,种子活力降低,种子萌发受抑制。