几种天然有机多酚酸体外抗脂质过氧化研究

通过天然有机多酚酸对H2O2诱导小鼠肝匀浆脂质过氧化和自发脂质过氧化的保护作用的体外实验,比较研究绿原酸、咖啡酸、迷迭香酸、阿魏酸、丹参素、原儿茶醛和3-羟基肉桂酸体外抗氧化能力的强弱,探讨其抗氧化能力产生机理及构效关系。结果表明,7种多酚酸对H2O2诱导小鼠肝匀浆脂质过氧化的保护作用强弱依次为迷迭香酸咖啡酸原儿茶醛绿原酸阿魏酸丹参素3-羟基肉桂酸,其最大和最小抑制率相差2.4倍;对小鼠肝脏组织自发脂质过氧化的抑制率大小顺序为迷迭香酸、原儿茶醛、绿原酸、咖啡酸、阿魏酸、丹参素、3-羟基肉桂酸,最高和最低抑制率相差近90%。7种天然有机酸抗脂质过氧化作用因化合物结构不同存在显著差异。     

藓类提取物对植物病原真菌的抑菌性研究

以立枯丝核菌(Rhizoctonia)、尖孢镰孢菌(Fusarium oxysporum)、苹果轮纹病菌(Macrophoma kawatsukai)、梨黑星病菌(Fusicladium pirina)、草莓灰霉菌(Botrytis cinerea)等七种植物病原真菌为供试菌,通过生长速率法测定了24种藓类的乙醇提取物对病原真菌的抑制作用.结果表明,在提取物浓度为1.2mg干样/mL,大镰刀藓提取物对立枯丝核菌的抑制作用最强,抑菌率为67.8%.藓类提取物不仅对植物病原真菌有抑菌活性,而且还有多种藓类提取物对病原真菌菌丝有促进生长活性,如锐尖匍灯藓对立枯丝核菌菌丝生长的促进率为95.1%.选出大镰刀藓、扁灰藓、紫萼藓、山地水灰藓、疣小金发藓、橙色净口藓、锐尖匍灯藓和羊角藓对立枯丝核菌和灰霉菌的抑菌性或菌丝生长促进性进行了进一步研究,藓类提取物对病原菌的抑菌性或菌丝生长促进性在一定范围内随浓度的增加而增强,并且抑菌性随时间的延长呈下降趋势.     

贝莱斯芽胞杆菌HN-Q-8菌株发酵液稳定性测定及抑菌活性成分分析

【背景】贝莱斯芽胞杆菌(Bacillus velezensis) HN-Q-8菌株是本实验室前期获得的能有效拮抗立枯丝核菌(Rhizoctonia solani)的生防细菌。【目的】明确贝莱斯芽胞杆菌HN-Q-8菌株的抑菌活性物质。【方法】采用菌丝生长速率法检测其发酵液对5种马铃薯病原菌的拮抗能力以及稳定性,利用基质辅助激光解吸电离飞行时间质谱技术对HN-Q-8菌株活性物质进行鉴定。【结果】HN-Q-8菌株对立枯丝核菌(Rhizoctonia solani)、尖镰孢菌(Fusarium oxysporum)、茄链格孢(Alternaria solani)和致病疫霉(Phytophthora infestans)具有良好的抑菌活性,抑菌率可达50%-90%。发酵液分别经紫外照射35min、自然光照射10 h以及100°C高温处理后,相对抑菌率分别为74%、92%和98%;发酵液的抑菌活性不受胰蛋白酶和蛋白酶K的影响;但不耐强酸、强碱,其适宜pH为4.0-10.0,表明HN-Q-8菌株活性物质具有良好的稳定性。活性成分能使立枯丝核菌菌丝形态畸形扭曲,从而抑制立枯丝核菌的生长,经鉴定活性物质为丰原素(fengycin)和表面活性素(surfactin)。【结论】贝莱斯芽胞杆菌HN-Q-8菌株发酵液具有较好的稳定性和较强的抑菌活性,具有良好的开发价值和应用前景。     

一株拮抗立枯丝核菌的放线菌筛选、鉴定及生理特性

通过对峙实验,从采集自黄渤海海域的双齿围沙蚕(Perinereis aibuhitensis)中分离到一株内生放线菌SCF-18,并对其抑菌活性进行分析。SCF-18对立枯丝核菌(Rhizoctonia solani)和小麦根腐病菌(Bipolaris sorokiniana)都具有拮抗作用,且对立枯丝核菌生长的拮抗效果最为明显,生长抑制率为89.78%。抑菌试验结果表明:菌株SCF-18发酵滤液可以抑制立枯丝核菌丝的生长,发酵液浓度越高,抑制力越强;当发酵液浓度达到50%时,对两种病原菌的抑制率分别为89.78%和80.26%。菌株SCF-18生长特性试验结果表明:菌株SCF-18生长最适温度为30℃,最适pH值为7;放线菌SCF-18耐盐度可达5%。根据形态特征、培养特征、生理生化特性和16S rDNA序列分析,将放线菌SCF-18初步鉴定为淡紫灰链霉菌(Streptomyces lavendulae)。本研究发现,SCF-18菌株是防治立枯丝核菌等病原菌的潜在优良生防菌株,具有潜在的开发应用价值。     

立枯丝核菌营养菌丝多型性观察

采用了2种不同的方法对立枯丝核菌营养菌丝的形态进行了观察和比较,观察到2种不同的营养菌丝的形态,即菌核类和假分生孢子类。为以后进一步研究立枯丝核菌营养菌丝的多型性奠定了一定的基础。     

叶损伤诱导兴安落叶松针叶中10种酚酸的变化

酚酸是一类重要次生抗虫物质.为研究损伤及昆虫取食诱导对兴安落叶松针叶内酚酸含量的影响,采用3种不同程度剪叶或落叶松毛虫幼虫取食处理兴安落叶松幼树,以高效液相色谱技术测定兴安落叶松健康针叶中酚酸含量.结果表明：与对照相比,处理后1 d,剪叶或昆虫取食4枝50%针叶处理的兴安落叶松幼苗健康针叶中,除阿魏酸无显著差异外,苯甲酸、咖啡酸、绿原酸、水杨酸、苯乙酸、肉桂酸、香草酸、丁香酸和没食子酸9种酚酸均差异显著;4枝75%针叶处理的10种酚酸含量均发生显著变化.说明剪叶及虫害50%、75%针叶处理均达到诱导阈值,能显著诱导兴安落叶松化学防御.在损伤程度相同情况下,处理1 d时,剪叶4枝50%、75%诱导的咖啡酸、苯乙酸、肉桂酸、香草酸和没食子酸的含量显著高于虫害诱导处理;5 d时,剪叶4枝50%、75%诱导处理的这5种酚酸含量显著低于虫害诱导处理;10 d时,两种方法诱导的酚酸含量差异不显著.说明剪叶诱导处理的酚酸含量变化比昆虫取食处理迅速,且诱导强度与剪叶程度相关.采用适当处理诱导针叶中酚酸含量的变化来增强兴安落叶松对害虫的防御能力是可行的.     

立枯丝核菌(Rhizoctonia solani)对日本结缕草(Zoysia japonica steud.)的侵染过程研究

【目的】了解立枯丝核菌(Rhizoctonia solani) AG1 IA对日本结缕草(Zoysia japonica steud.)的侵染过程及其引起的病害症状,为进一步从分子水平研究该菌的病理学侵染机制和草坪草抗病分子育种提供理论基础。【方法】通过根部接种法促使立枯丝核菌与日本结缕草无菌苗建立侵染关系,从而对其病叶率、病株率及病情指数进行统计分析,同时结合组织染色透明技术及植物组织石蜡切片对立枯丝核菌的侵染过程、感染方式进行研究。【结果】立枯丝核菌AG1 IA的侵染过程主要为：菌丝吸附在植物组织表面,并沿组织表面定向生长,形成侵染结构——侵染垫与组织建立密切的侵染关系。菌丝通过细胞间隙侵入植物组织内部,主要侵染植物皮层细胞及除木质部导管以外的整个维管束系统。结缕草的地上部分与地下部分组织对立枯丝核菌的侵染显现不同的寄主反应。【结论】立枯丝核菌的侵染过程主要包括吸附、定向生长、渗透、定殖4个部分;立枯丝核菌的侵染主要引起结缕草叶片病症;结缕草病变与菌丝直接侵染无直接联系,表明该菌具有复杂的侵染机制。     

河西走廊盐碱土壤中抗立枯丝核菌的放线菌筛选

从甘肃省河西走廊盐碱土中分离所得50株放线菌中,筛选出对立枯丝核菌有生防潜能的放线菌,旨在为生物防治作物病害提供新的菌种资源。通过平板对峙法、生长速率法及活体组织法筛选拮抗菌,并对其进行形态及分子鉴定。经过平板初筛,有4株菌株对立枯丝核菌有拮抗作用,编号Ⅰ18-5-2、Ⅲ22-3-12、Ⅳ16-3-2、DC009-1-23,其中Ⅳ16-3-2和DC009-1-23有较高的拮抗性,抑菌圈直径分别为20.5 mm和15.5 mm;进而对这2菌株进行发酵液复筛,菌丝的生长抑制率分别为89.02%和80.49%;活体组织试验表明,Ⅳ16-3-2和DC009-1-23对立枯丝核菌的防治效果分别达54.29%和45.71%;通过形态培养特征和16S rDNA序列鉴定,将菌株Ⅳ16-3-2初步鉴定为变异链霉菌,DC009-1-23为丁香链霉菌。     

两种秸秆醋液对尖孢镰刀菌和立枯丝核菌的抑菌作用

本研究以玉米秸秆醋液、水稻秸秆醋液为供试材料,研究不同稀释度的秸秆醋液对尖孢镰刀菌(Fusarium oxysporum)和立枯丝核菌(Rhizoctonia solani)的抑制作用,测定其可溶性蛋白和可溶性糖。结果表明,50倍稀释液对2种病原菌的抑制率为100%,即2种病原菌在50倍稀释处理的秸秆醋液中不能生长。玉米秸秆醋液对尖孢镰刀菌中可溶性蛋白的影响不显著,对立枯丝核菌中可溶性蛋白的影响表现为先升高后降低。水稻秸秆醋液对尖孢镰刀菌中可溶性蛋白的影响是先升高后降低,对立枯丝核菌中可溶性蛋白的影响是逐渐累积。高浓度的水稻秸秆醋液可提高2种菌体内可溶性糖含量,100倍稀释的玉米秸秆醋液可提高尖孢镰刀菌体内的可溶糖含量,玉米秸秆醋液对立枯丝核菌的可溶性糖含量基本不会产生影响。本研究为秸秆醋液在农业生产上的推广应用等提供科学依据。     

酚酸类物质的抑草效应分析

运用正交旋转回归试验设计分析5种常见的化感物质替代物水饧酸、对羟基苯甲酸、肉桂酸、香草酸和阿魏酸对田间伴生杂草稗草的抑制效应．结果表明,肉桂酸对稗草根长抑制率的影响最显著。其关系函数的二次项系数为-6．18,达极显著水平,水杨酸、对羟基苯甲酸和阿魏酸对稗草根长的抑制效应趋势与肉桂酸相同,效应曲线均为“n”形抛物线;而香草酸的效应曲线则为“U”形抛物线．当水饧酸、对羟基苯甲酸、肉桂酸、香草酸和阿魏酸浓度水平分别为0．06、0．60、0．24、0．02和0．02mmol·L^-1时,混合物对稗草根长的抑制率最大,达到78．65％。     

Interaction of Moisture Stress and Three Phenolic Compounds on Lettuce Seed Germination

No interactions between water stress and three phenolic acids(p-coumaric, caffeic and ferulic acids) on lettuce (Lactucasativa L. var. Grand Rapids) seed germination were found. Probitanalysis indicated that mechanisms of action of water stressand the phenolic inhibitors were similar. The relative effectivenessof the compounds was p-coumaric > ferulic > caffeic. Nointeraction was found between p-coumaric and ferulic acid, whereasantagonism was found between caffeic acid and each of the othertwo phenolic acids. Lactuca sativa L., lettuce, germination, phenolic compounds, moisture stress, allelopathy, seed     

Belowground ABA boosts aboveground production of DIMBOA and primes induction of chlorogenic acid in maize

Plants are important mediators between above- and belowground herbivores. Consequently, interactions between root and shoot defenses can have far-reaching impacts on entire food webs. We recently reported that infestation of maize roots by larvae of the beetle Diabrotica virgifera virgifera induced shoot resistance against herbivores and pathogens. Root herbivory also enhanced aboveground DIMBOA and primed for enhanced induction of chlorogenic acid, two secondary metabolites that have been associated with plant stress resistance. Interestingly, the plant hormone abscisic acid (ABA) emerged as a putative long-distance signal in the regulation of these systemic defenses. In this addendum, we have investigated the role of root-derived ABA in aboveground regulation of DIMBOA and the phenolic compounds chlorogenic acid, caffeic and ferulic acid. Furthermore, we discuss the relevance of ABA in relation to defense against the leaf herbivore Spodoptera littoralis. Soil-drench treatment with ABA mimicked root herbivore-induced accumulation of DIMBOA in the leaves. Similarly, ABA mimicked aboveground priming of chlorogenic acid production, causing augmented induction of this compound after subsequent shoot attack by S. littoralis caterpillars. These findings confirm our notion that ABA acts as an important signal in the regulation of aboveground defenses during belowground herbivory. However, based on our previous finding that ABA alone is not sufficient to trigger aboveground resistance against S. littoralis caterpillars, our results also suggest that the ABA-inducible effects on DIMBOA and chlorogenic acid are not solely responsible for root herbivore-induced resistance against S. littoralis.Key words: induced resistance, Spodoptera littoralis, Zea mays, Diabrotica virgifera, DIMBOA, chlorogenic acid, absisic acid, priming     

Antimutagenic effect of phenolic acids

In the present study, the Salmonella typhimurium tester strain TA 100 was used in the plate-incorporation test to examine the antimutagenic potential of caffeic, ferulic and cichoric acids extracted from plant species of genera Echinacea (L) Moench, as well as of another phenolic acids, on 3-(5-nitro-2-furyl)acrylic acid (5NFAA) and sodium azide mutagenicity. All tested compounds possess antimutagenic activity. In the case of 5NFAA, the antimutagenic potency of tested compounds was in the order of gallic acid > ferulic acid > caffeic acid > syringic acid > vanillic acid. The mutagenic effect of sodium azide was inhibited by tested phenolic acids by about 20-35 %. The most effective compound, gallic acid inhibits this effect by 82 % in the concentration of 500 mug/plate. The only exception from favourable properties of tested phenolic acids is cichoric acid, which in the contrary significantly increased the mutagenic effect of 5NFAA.     

Biochemical features of maize tissues with different capacities to regenerate plants

Metabolic profiling using GC–MS and LC–MS analyses of soluble metabolites and cell wall bound phenolic compounds from maize calluses of different morphogenic competence revealed a number of biochemical characteristics that distinguish tissues with high plant regeneration ability from tissues that cannot efficiently regenerate plants in vitro. Maize cultures of different ages from H99 (compact type I callus) and HiII (friable type II callus) were divided into two different samples: regenerable (R) and non-regenerable (NR) based on known morphologies. Tissues from both genotypes with high morphogenic potential had higher asparagine and aspartate and indole-3-butenol concentrations, decreased sugar and DIMBOA (2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one) concentrations, low levels of 4-aminobutyric acid (GABA) and chlorogenic acid and lower levels of feruloyl- and sinapoyl glucosides compared to NR tissues. The ether bound cell wall phenolics of tissues with high regeneration potential had higher levels of the predominant G (guaiacyl) units and lower levels of H (p-hydroxyphenyl) and S (syringyl) units and higher ferulic acid/coumaric acid and ferulic acid/diferulic acid ratios. The same trends were found with the ester-bound phenolics of HiII, however, there were only small differences between the H99 R and NR tissues. Concentrations of the major sugars, organic acids, amino acids and soluble aromatic compounds tended to increase as the time after culture initiation increased. The results show that there are differences in general metabolism, phenolic secondary compounds and cell wall composition between R and NR cell types.     

The phenolic vir gene inducer ferulic acid is O-demethylated by the VirH2 protein of an Agrobacterium tumefaciens Ti plasmid

Some or possibly all Ti plasmids of Agrobacterium tumefaciens encode a bicistronic operon designated virH, which encodes two proteins, VirH1 and VirH2, that resemble a family of cytochrome P450-type monooxygenases. Expression of this operon is induced by a family of phenolic compounds that induce all other operons within the vir regulon. We hypothesized that either or both of these proteins might metabolize some or all of these phenolic compounds. We therefore tested induction of a vir promoter by a variety of phenolic compounds in isogenic strains that express or lack virH1 and virH2. Although some compounds were equally effective inducers regardless of the virH status, other compounds induced vir expression far more effectively in the virH mutant than in the virH-proficient host. For all tested compounds, VirH2 appeared to be solely responsible for this effect. One such compound, ferulic acid, was chosen for biochemical analysis. Ferulic acid was degraded by a VirH-proficient host but not by a VirH mutant. The wild-type strain released large amounts of a more hydrophilic compound into the cell supernatant. This compound was tested by mass spectroscopy, nuclear magnetic resonance and UV spectroscopy and found to consist of caffeic acid. This indicates that wild-type strains convert virtually all added ferulic acid to caffeic acid, and that VirH2 is essential for this O-demethylation reaction. Ferulic acid was far more toxic than caffeic acid to the wild-type strain, although the wild-type strain was more resistant to ferulic acid than was the virH mutant. Caffeic acid was slowly removed from the broth, suggesting further metabolic reactions.     

藏药翁布的酚性成分研究

从藏药翁布(Myricaria germanica)的60％丙酮提取物中进行了进一步研究,从中共分离得到了11个化合物.利用光谱和波谱分析法,分别鉴定为阿魏酸(1),松柏醇(2),阿魏酸葡糖苷(3),异落叶松脂醇(4),咖啡酸(5),对羟基桂皮酸(6),没食子酸(7),3,5-二羟基-4-甲氧基苯甲酸(8),杜鹃醇(9),3-甲氧基-4-羟基-苯甲酸(10)和3,4,5-三羟基肉桂酸(11).化合物1～11均为首次从该植物中分得,其中1～6和8～11为首次从水柏枝属植物中分离得到.     

Effects of phenolic acids on ammonia oxidation by terrestrial autotrophic nitrifying microorganisms

Abstract It has been hypothesized that vegetation in certain ecosystems inhibits nitrification in soil by producing phenolic compounds that inhibit oxidation of ammonia by nitrifying microorganisms. This hypothesis is based largely on a report that very low concentrations (10⁻⁶ M–10⁻⁸ M) of several phenolic acids (notably ferulic acid) completely inhibited NO₂⁻ production in an aqueous suspension of soil treated with (NH₄)₂SO₄ and a nutrient solution suitable for growth of Nitrosomonas and other autotrophic nitrifying microorganisms. To evaluate this hypothesis, we determined the effects of three ohenolic acids (ferulic acid, caffeic acid, and p -coumaric on nitrite production by representatives of three genera of terrestrial autotrophic nitrifying microorganisms ( Nitrosospira, Nitrosomonas , or Nitrosolubos ) grown on a defined medium containing NH₄⁺. We found that nitrite production by the Nitrososspira was not inhibited by ferulic acid, caffeic acid, or p -coumaric acid at concentrations of 10⁻⁶ or 10⁻⁵ M and was only slightly inhibited when these acids were at a concentration of 10⁻⁴ M. We also found that ferulic acid did not markedly inhibit nitrite production by the three genera of nitrifying microorganisms studied, even when its concentration was as high as 10⁻³ M. These observations invalidate the hypothesis tested because the phenolic acids studied did not significantly retard ammonia oxidation by autotrophic microorganisms even when their concentration in cultures of these microorganisms greatly exceeded their concentrations in soils.     

Free phenols in maize pith and their relationship with resistance to Sesamia nonagrioides (Lepidoptera: Noctuidae) attack

The stem borer Sesamia nonagrioides (Lefèbvre) is the most important insect pest of maize, Zea mays L., in northwestern Spain. Among the metabolites present in maize, phenolic compounds could play an important role in resistance. The objective of this work was to determine whether a relationship between phenols and the amount of resistance exists. Amounts of free phenolic compounds in the pith of 13 inbred maize lines that differ in resistance were measured. The phenolic compounds identified were p-coumaric acid, cafeic acid, ferulic acid, vanillic acid, syringic acid, chorogenic acid, sinapic acid, p-hydroxybenzoic acid, and vanillin. The amount of free p-coumaric acid was correlated with the resistance level. Higher quantities of p-coumaric in the pith could contribute to general resistance to stem borer attack. Jointly with ferulic acid, p-coumaric could provide resistance mechanisms through cell wall fortification and lignification. The other compounds showed no or an unclear relationship with resistance. The vanillic acid showed a decreased tendency after silking, when maize is most attractive for S. nonagrioides, suggesting this acid could act as a chemoattractant for S. nonagrioides larvae or adults. Future studies that focus on these phenolic compounds could be useful in understanding S. nonagrioides resistance.     

The effect of hydroxycinnamic acids on the growth of wine-spoilage lactic acid bacteria

Hydroxycinnamic acids and their derivatives occur naturally in grape juice and wine. To assess their potential as natural preservatives the effect of caffeic, coumaric and ferulic acids on the growth of three wine-spoilage strains of Lactobacillus collinoides and one of Lact. brevis was studied in acid tomato broth containing 5% ethanol at pH 4.8. At concentrations of 500 and 1000 mg l^-1, all three compounds markedly inhibited growth; coumaric and ferulic acids were more effective than caffeic acid. At a concentration of 100 mg l^-1, all compounds stimulated growth. In general, the strains of Lact. collinoides were more susceptible both to inhibition and stimulation by the hydroxycinnamic acids than was the strain of Lact. brevis. The possible influence of hydroxycinnamic acids on the malolactic fermentation of wine is discussed.     

Phenolic compounds and somatic embryogenesis in cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis> L.)

Studies of phenolic compounds were performed during cell suspension cultures in relation with the induction of embryogenic structures in two cultivars of cotton. Coker 312 produced embryogenic structures, unlike R405-2000 which was found to be a non-embryogenic cultivar. Embryogenesis induction in Coker 312 was strongly linked to a higher content of caffeic, ferulic and salicylic acids and to the appearance of p-coumaric acid, benzoic acid, trans-resveratrol, catechin and naringenin.