Effect of Plant Growth-Promoting Rhizobacteria and Culture Filtrate of <Emphasis Type="Italic">Sclerotium rolfsii</Emphasis> on Phenolic and Salicylic Acid Contents in Chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis>)

Two plant growth-promoting rhizobacteria (PGPR), viz., Pseudomonas fluorescens strain Pf4 and P. aeruginosa strain Pag, protected chickpea (Cicer arietinum) plants from Sclerotium rolfsii infection when applied singly or in combination as seed treatment. Pag gave the best protection to the seedlings, applied either singly (mortality 16%) or in combination with Pf4 (mortality 17%) compared with 44% and 24% mortality in control and Pf4 treatment, respectively. The two PGPR strains induced the synthesis of specific phenolic acids, salicylic acid (SA), as well as total phenolics at different growth stages of chickpea seedlings with varied amount. The maximum amount of total phenolics was recorded in all the aerial parts of 4-week-old plants. Gallic, ferulic, chlorogenic, and cinnamic acids were the major phenolic acids detected in high-performance liquid chromatography (HPLC) analysis. Induction of such phenolic acids in the seedlings was observed up to 6 weeks in comparison with control. Salicylic acid (SA) was induced frequently during the first 3 weeks of growth only. Between the two strains, Pag was more effective in inducing phenolic acid synthesis applied either singly or in combination with strain Pf4 during the entire 6 weeks of growth of chickpea. In the presence of a culture filtrate of S. rolfsii, the two Pseudomonas strains induced more phenolic acids in treated than in non-treated and control plants. The occurrence of salicylic acid was frequent in the first 24 h, but infrequent at 48 and 96 h. Foliar spray of Pseudomonas strains also enhanced the phenolic acid content as well as total phenolics within 24 h of application. Gallic, chlorogenic, and cinnamic acids were consistently discerned in the treated leaves, whereas SA was absent even up to 96 h of application. Resistance in chickpea plants by Pseudomonas strains through induction of phenolic compounds as well as induced systemic resistance via SA-dependent pathway was evident. Received: 1 April 2002 / Accepted: 4 May 2002     

Rhizobium-Mediated Induction of Phenolics and Plant Growth Promotion in Rice (Oryza sativa L.)

Qualitative and quantitative estimation of phenolic compounds was done through reverse phase–high performance liquid chromatography (RP-HPLC) from different parts (leaf, stem, and root) of rice plants after inoculation with two rhizobial strains, RRE6 (Rhizobium leguminosarum bv. phaseoli) and ANU 843 (R. leguminosarum bv. trifolii) and infection by Rhizoctonia solani. On the basis of their retention time, the major phenolic acids detected in HPLC analysis were gallic, tannic, ferulic, and cinnamic acids. Furthermore, in all Rhizobium-inoculated rice plants, synthesis of phenolic compounds was more consistently enhanced than in control (uninoculated plants), where the maximum accumulation of phenolic compounds was observed in plants inoculated with RRE6 and infection with R. solani. Under pathogenic stress, RRE6 performed better because a relatively higher amount of phenolics was induced as compared with plants treated with ANU 843. Phenolic acids mediate induced systemic resistance and provide bioprotection to plants during pathogenic stresses. In addition, both rhizobial strains promote growth and productivity of rice plants in greenhouse conditions. This report on Rhizobium-mediated defense responses and growth promotion of nonlegume (such as rice) provides a novel paradigm of symbiotic plant–microbe interaction.     

Effect of plant growth-promoting Rhizobacteria and culture filtrate of Sclerotium rolfsii on phenolic and salicylic acid contents in chickpea (Cicer arietinum)

Two plant growth-promoting rhizobacteria (PGPR), viz., Pseudomonas fluorescens strain Pf4 and P. aeruginosa strain Pag, protected chickpea ( Cicer arietinum) plants from Sclerotium rolfsii infection when applied singly or in combination as seed treatment. Pag gave the best protection to the seedlings, applied either singly (mortality 16%) or in combination with Pf4 (mortality 17%) compared with 44% and 24% mortality in control and Pf4 treatment, respectively. The two PGPR strains induced the synthesis of specific phenolic acids, salicylic acid (SA), as well as total phenolics at different growth stages of chickpea seedlings with varied amount. The maximum amount of total phenolics was recorded in all the aerial parts of 4-week-old plants. Gallic, ferulic, chlorogenic, and cinnamic acids were the major phenolic acids detected in high-performance liquid chromatography (HPLC) analysis. Induction of such phenolic acids in the seedlings was observed up to 6 weeks in comparison with control. Salicylic acid (SA) was induced frequently during the first 3 weeks of growth only. Between the two strains, Pag was more effective in inducing phenolic acid synthesis applied either singly or in combination with strain Pf4 during the entire 6 weeks of growth of chickpea. In the presence of a culture filtrate of S. rolfsii, the two Pseudomonas strains induced more phenolic acids in treated than in non-treated and control plants. The occurrence of salicylic acid was frequent in the first 24 h, but infrequent at 48 and 96 h. Foliar spray of Pseudomonas strains also enhanced the phenolic acid content as well as total phenolics within 24 h of application. Gallic, chlorogenic, and cinnamic acids were consistently discerned in the treated leaves, whereas SA was absent even up to 96 h of application. Resistance in chickpea plants by Pseudomonas strains through induction of phenolic compounds as well as induced systemic resistance via SA-dependent pathway was evident.     

Antibiotic potential of plant growth promoting rhizobacteria (PGPR) against Sclerotium rolfsii

High performance liquid chromatographic (HPLC) analysis of culture filtrates of plant growth promoting rhizobacteria (PGPR) and medium of inhibitory zone of interaction of Sclerotium rolfsii with PGPR, viz. Pseudomonas aeruginosa, Pseudomonas fluorescens 4, Pseudomonas fluorescens 4 (new) and Pseudomonas sp. varied from sample to sample. In all the culture filtrates of PGPRs, P. aeruginosa had nine phenolic acids in which ferulic acid (14.52 μg/ml) was maximum followed by other phenolic acids. However, the culture filtrates of P. fluorescens 4 had six phenolic acids with maximum ferulic acid (20.54 μg/ml) followed by indole acetic acid (IAA), caffeic, salicylic, o-coumeric acid and cinnamic acids. However, P. fluorescens 4 culture filtrate had seven phenolic acids in which salicylic acid was maximum (18.03 μg) followed by IAA, caffeic, vanillic, ferulic, o-coumeric and cinnamic acids. Pseudomonas sp. also showed eight phenolic acids where caffeic acid (2.75 μg) was maximum followed by trace amounts of ferulic, salicylic, IAA, vanillic, cinnamic, o-coumeric and tannic acids. The analysis of antibiosis zone of PGPRs showed fairly rich phenolic acids. A total of nine phenolic acids were detected in which caffeic acid was maximum (29.14 μg/g) followed by gallic (17.64 μg/g) and vanillic (3.52 μg/g) acids but others were in traces. In P. aeruginosa, antibiosis zone had seven phenolic acids where IAA was maximum (3.48 μg/g) followed by o-coumeric acid (2.08 μg/g), others were in traces. The medium of antibiosis zone of P. fluorescens 4 and P. fluorescens 4 new had eight phenolic acids in which IAA was maximum with other phenolic acids in traces.     

Phenolic compounds of Sorghum vulgare in response to Sclerotium rolfsii infection

Abstract

Identification of individual phenolic acids of Sorghum vulgare Pers. cv. M.P. after interaction with Sclerotium rolfsii Sacc. using high performance liquid chromatograph (HPLC) showed the presence of phenolics namely tannic, gallic, ferulic, chlorogenic and cinnamic acids in varying amounts. After 72 h inoculation with S. rolfsii, a maximum amount of ferulic acid (166.6 µg g^?1 fresh wt) was present in the collar of inoculated plants, followed by leaves and roots and its level decreased gradually with time. Similarly, the presence of chlorogenic acid was traced after 48 h, while that of cinnamic acid was traced after 72 h of inoculation. Reddish-brown pigmentation at the collar region of inoculated plants was also observed along with the high content of tannic acid. Among other phenolics, the presence of gallic acid was recorded consistently and maximum accumulation (139.3 µg g^?1 fresh wt) was noticed at the zone of interaction (collar region) after 72 h of inoculation. In contrast, maximum lignin deposition was observed at collar region after 96 h of inoculation. Induction of phenolic acids in S. vulgare along with the lignin deposition and red pigmentation at collar region is considered a key biomarker in the non-host-pathogen interaction in the S. valgare–S. rolfsii pathosystem.     

Studies on exudate-depleted sclerotial development in Sclerotium rolfsii and the effect of oxalic acid,sclerotial exudate,and culture filtrate on phenolic acid induction in chickpea (Cicer arietinum)

Exudate depletion from developing sclerotia of Sclerotium rolfsii Sacc. in culture caused reduced size and weight of sclerotia. Germination of exudate-depleted sclerotia was delayed on Cyperus rotundus rhizome meal agar medium when compared with that of control sclerotia. The exudate-depleted sclerotia caused infection in chickpea (Cicer arietinum) plants in a glasshouse. Different temperatures and incubation periods had no effect on the germination ability of the exudate-depleted sclerotia. Oxalic acid, sclerotial exudate, and culture filtrate of S. rolfsii induced the synthesis of phenolic acids, including gallic, ferulic, chlorogenic, and cinnamic acids, as well as salicylic acid, in treated chickpea leaves. Gallic acid content was increased in treated leaves compared with the untreated controls. Maximum induction of gallic acid was seen in both leaves treated with oxalic acid followed by exudate and leaves treated with culture filtrate. Cinnamic and salicylic acids were not induced in exudate-treated leaves. Ethyl acetate fractionation indicated that the sclerotial exudates consisted of gallic, oxalic, ferulic, chlorogenic, and cinnamic acids, whereas the culture filtrate consisted of gallic, oxalic, and cinnamic acids along with many other unidentified compounds.     

Biochemical Investigations of Sclerotial Exudates of Sclerotium rolfsii and their Antifungal Activity

Exudates from sclerotia of two Sclerotium rolfsii isolates (one causing collar rot in Cicer arietinum, isolate VC971, and the other leaf spots in Rauvolfia serpentina, isolate VL016) were assayed for their antifungal activity against 26 fungi consisting of plant parasites as well as saprophytes. Spore germination of all the test fungi was affected by the exudates reaching 100% in some cases. Foliar spray with exudates of isolate VL016 significantly reduced disease incidence of balsam (Impatiens balsamina) powdery mildew caused by Erysiphe cichoracearum and pea (Pisum sativum) powdery mildew caused by Erysiphe pisi, under field conditions. Characterization of exudates from 25 isolates of S. rolfsii revealed pH ranging from 3.8 to 5.3 and colour from light yellow to deep yellow. Among the phenolic acids found in the exudates were tannic, gallic, caffeic, vanillic, ferulic, chlorogenic and cinnamic acids. Oxalic acid was also found in varied amounts. Among the phenolic acids, ferulic acid was found to be present at high concentration in exudates of most isolates (3.9–153.4 μg/ml). The antioxidant properties of phenolics, which generally inhibit fungal morphogenesis including spore germination along with the antifungal nature of some phenolics are chiefly attributed to the inhibitory effect of sclerotial exudates of S. rolfsii. Additionally, both the isolates VC971 and VL016 showed almost similar antifungal activities despite they are of different origin and thereby demonstrate the antifungal nature of sclerotial exudates.     

甜瓜根系分泌物中酚酸物质对尖孢镰孢菌的化感效应

采用HPLC法对甜瓜根系分泌物进行分离鉴定,检测到甜瓜根系分泌物中含有没食子酸、邻苯二甲酸、丁香酸、水杨酸、阿魏酸、苯甲酸和肉桂酸7种酚酸物质,通过外源添加法研究该类物质对尖孢镰孢菌的化感效应.室内试验结果表明: 阿魏酸、苯甲酸、肉桂酸在0.1、0.25 mmol·L^-1处理浓度下能够显著促进尖孢镰孢菌的孢子萌发,水杨酸则对孢子萌发具有一定的抑制作用;丁香酸、阿魏酸在菌丝培养后期表现出较强的促进作用.盆栽结果显示,在0.05、0.1和0.5 mmol·L^-1处理浓度下肉桂酸、阿魏酸、苯甲酸可显著促进甜瓜枯萎病病情.
     

Exogenous application of L-phenylalanine and ferulic acid enhance phenylalanine ammonia lyase activity and accumulation of phenolic acids in pea (Pisum sativum) to offer protection against Erysiphe pisi

Phenylalanine ammonia lyase (PAL) activity was measured using HPLC in pea leaves following exogenous application of L-phenylalanine and ferulic acid. Treatment with different concentrations (50, 100, 150 ppm) of L-phenylalanine caused increased activity of PAL activity in comparison to control. In pea leaves treated with 50 ppm L-phenylalanine, maximum PAL activity was observed after 72 h of treatment. Application of ferulic acid first reduced PAL activity at lower concentration (50 ppm) but it further increased at higher concentrations of the compound (100 and 150 ppm) in pea leaves compared to control. Minimum PAL activity was 0.19 nM cinnamic acid/min/g fresh wt after 24 h at 50 ppm and then increased with time. Treatment with both compounds significantly increased the accumulation of phenolic acids and salicylic acid and reduced conidial germination of Erysiphe pisi on pea leaves. They were equally effective at 100 and 150 ppm in reducing conidial germination. Conidial germination on L-phenylalanine-treated leaves was 26% after 24 h and that on ferulic acid treated leaves 34% compared to control (46%). Foliar application of different concentrations of L-phenylalanine increased the level of ferulic acid in the leaves of pea plants. Maximum enzyme activity in terms of the accumulation of cinnamic acid (79.3 and 83.5 μg/g fresh wt) was observed following the application of L-phenylalanine after 24 and 48 h respectively. At 50 ppm, cinnamic acid accumulation in pea leaves was 35.6 and 39.4 μg/g fresh wt and 74.3 and 86.5 μg/g fresh wt at 100 ppm.     

Induction of Plant Defense Enzymes and Phenolics by Treatment With Plant Growth–Promoting Rhizobacteria Serratia marcescens NBRI1213

In greenhouse experiments, plant growth–promoting rhizobacteria (PGPR) Serratia marcescens NBRI1213 was evaluated for plant growth promotion and biologic control of foot and root rot of betelvine caused by Phytophthora nicotianae. Bacterization of betelvine (Piper betle L.) cuttings with S. marcescens NBRI1213 induced phenylalanine ammonia-lyase, peroxidase, and polyphenoloxidase activities in leaf and root. Qualitative and quantitative estimation of phenolic compounds was done through high-performance liquid chromatography (HPLC) in leaf and root of betelvine after treatment with S. marcescens NBRI1213 and infection by P. nicotianae. Major phenolics detected were gallic, protocatechuic, chlorogenic, caffeic, ferulic, and ellagic acids by comparison of their retention time with standards through HPLC. In all of the treated plants, synthesis of phenolic compounds was enhanced compared with control. Maximum accumulation of phenolics was increased in S. marcescens NBRI1213–treated plants infected with P. nicotianae. In a greenhouse test, bacterization using S. marcescens NBRI1213 decreased the number of diseased plants compared with nonbacterized controls. There were significant growth increases in shoot length, shoot dry weight, root length, and root dry weight, averaging 81%, 68%, 152%, and 290%, respectively, greater than untreated controls. This is the first report of PGPR–mediated induction of phenolics for biologic control and their probable role in protecting betelvine against P. nicotianae, an important soil-borne phytopathogenic fungus.     

芦苇腐解土中酚酸类化感物质的水分响应特性研究

芦苇和虉草均具有较强的去污能力,常作为湿地植物配置于同一人工湿地进行污水处理。芦苇作为一种强化感植物对虉草具有较强的化感作用,在自然湿地和人工湿地中均会出现芦苇代替虉草的现象,且这一现象的发生与土壤含水量存在一定联系,此外,芦苇腐解土对虉草的化感抑制效应与腐解土中总酚酸的量密切相关。为了研究芦苇腐解土中主要酚酸类物质的水分响应特性,筛选出其中对水分响应较为明显的酚酸物质种类,该研究采用高效液相色谱法,通过芦苇枯落物腐解土的制备,对不同水分环境下芦苇腐解土中酚酸类物质进行了分离和鉴定。结果表明:芦苇腐解土中可分离出没食子酸、香豆酸、香草酸、丁香酸、对香豆酸、阿魏酸、水杨酸和苯甲酸等8种酚酸类物质,其中香豆酸、苯甲酸和阿魏酸等3种酚酸类物质含量较高。分离出的8种酚酸类物质的含量与腐解土的相对含水量均呈显著线性负相关关系,即随着腐解土相对含水量的上升,酚酸类物质的含量均呈现下降趋势,且各种酚酸类物质对水分的响应趋势均可用线性方程较好地拟合。其中,香豆酸、没食子酸和阿魏酸对芦苇腐解土的水分响应最为明显。因此,可将香豆酸、没食子酸和阿魏酸作为主要调控目标,通过调控湿地土壤中水分含量,削弱芦苇对虉草的化感抑制效应,从而维持人工湿地中虉草芦苇群落的长期稳定共存。     

Ferulic acid may prevent infection of Cicer arietinum by Sclerotium rolfsii

High performance liquid chromatography analysis of different parts of Sclerotium rolfsii-infected and healthy seedlings of chickpea (Cicer arietinum) was carried out to examine the status of phenolic compounds. Three major peaks that appeared consistently were identified as gallic, vanillic and ferulic acids. Gallic acid concentrations were increased in the leaves and stems of infected plants compared to healthy ones. Vanillic acid detected in stems and leaves of healthy seedlings was not detected in infected seedlings. There was a significant increase of ferulic acid in those stem portions located above the infected collar region compared to minimal amounts in the roots of healthy seedlings. In vitro studies of ferulic acid showed significant antifungal activity against S. rolfsii. Complete inhibition of mycelial growth was observed with 1000 g of ferulic acid/ml. Lower concentrations (250, 500 and 750 g/ml) were also inhibitory and colony growth was compact in comparison with the fluffy growth of normal mycelium. Higher amounts of phenolics were found in the stems and leaves of S. rolfsii-infected seedlings in comparison to the healthy ones. A role for ferulic acid in preventing infections by S. rolfsii in the stems and leaves of chickpea plants above the infection zone is therefore feasible.     

Antioxidant activity of phenolic and related compounds: a density functional theory study on the O–H bond dissociation enthalpy

Abstract

We report here on calculations at the hybrid DFT/HF (B3-LYP/6-31G(d, p)) level of the O–H bond dissociation enthalpy (O–H BDE) of phenylpropenoic acids (caffeic, ferulic, p-coumaric and cinnamic) and phenolic acids and related compounds (gallic, methylgallate, vanillic and gentisic) in order to gain insight into the understanding of structure–antioxidant activity relationships. The results were correlated and discussed mainly on the basis of experimental data in a companion work (Galato D, Giacomelli C, Ckless K, Susin MF, Vale RMR, Spinelli A. Antioxidant capacity of phenolic and related compounds: correlation among electrochemical, visible spectroscopy methods and structure-antioxidant activity. Redox Report 2001; 6: 243–250). The O–H BDE values showed remarkable dependence on the hydroxyl position in the benzene ring and the existence of additional interaction due to hydrogen bonding. For parent molecules, the experimental antioxidant activity (AA) order was properly obeyed only when intramolecular hydrogen bonding was present in the radicalized structures of o-dihydroxyl moieties. In structurally related compounds, excellent correlation with experimental data was in general observed (0.64 < ρ < 0.99). However, it is shown that excellent correlation can also be obtained for this series of compounds considering p-radicalized structures which were not stabilized by intramolecular hydrogen bonding, but this had no physical meaning. These findings suggested that the antioxidant activity evaluation of phenolic and related compounds must take into consideration the characteristics of each particular compound.     

Antioxidant Activity,Total Phenolic,Tannin, and Flavonoid Content of Five Plants Used in Traditional Medicine in Penjamo,Guanajuato

In Mexico, plants are commonly used to alleviate various ailments, including controlling some chronic degenerative diseases through the regular consumption of decoctions, infusions, and teas. However, there is little scientific evidence consolidating traditional medicine within health systems. Therefore, this work determined the phytochemical profile of the most used plants to treat various ailments (Cedro rojo, Cancerina, Ortiguilla, Hierba de la golondrina, Hierba de arlomo) and their general consumption as infusions. Aqueous and ethanolic extracts were generated, while the phytochemical compound content in the extracts obtained was quantified. The results indicate that the ethanolic extracts showed the highest phenolic compound and tannin content, with the highest contents for Cedro rojo (831.04 mg L⁻¹) and Cancerina (864.80 mg L⁻¹). The antioxidant activity was also determined, and a significant difference was observed (p<0.05). The extracts with the highest antioxidant capacity were the ethanolic extracts ranging from 250 to 907 μMET mL⁻¹, while the aqueous extracts ranged from 112 to 390 μMET mL⁻¹. The compounds identified by high-performance liquid chromatography characterization on the aqueous extracts highlighted the presence of chlorogenic acid>cinnamic acid>quercetin. In ethanolic extracts, the presence of chlorogenic acid>cinnamic acid>quercetin>gallic acid>ferulic acid>coumaric acid was highlighted. The correlation between bioactive compounds, type of extract, and antioxidant activity suggests a significant affinity of these phytochemical compounds for the ethanol solvent. The results indicate that these plants are good sources of antioxidant phenolics and can be incorporated for use as functional beverages. However, more studies are needed to corroborate their beneficial effect.     

Cyanobacteria-mediated phenylpropanoids and phytohormones in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis>) enhance plant growth and stress tolerance

Phenylpropanoids, flavonoids and plant growth regulators in rice (Oryza sativa) variety (UPR 1823) inoculated with different cyanobacterial strains namely Anabaena oryzae, Anabaena doliolum, Phormidium fragile, Calothrix geitonos, Hapalosiphon intricatus, Aulosira fertilissima, Tolypothrix tenuis, Oscillatoria acuta and Plectonema boryanum were quantified using HPLC in pot conditions after 15 and 30 days. Qualitative analysis of the induced compounds using reverse phase HPLC and further confirmation with LC-MS/MS showed consistent accumulation of phenolic acids (gallic, gentisic, caffeic, chlorogenic and ferulic acids), flavonoids (rutin and quercetin) and phytohormones (indole acetic acid and indole butyric acid) in rice leaves. Plant growth promotion (shoot, root length and biomass) was positively correlated with total protein and chlorophyll content of leaves. Enzyme activity of peroxidase and phenylalanine ammonia lyase and total phenolic content was fairly high in rice leaves inoculated with O. acuta and P. boryanum after 30 days. Differential systemic accumulation of phenylpropanoids in plant leaves led us to conclude that cyanobacterial inoculation correlates positively with plant growth promotion and stress tolerance in rice. Furthermore, the study helped in deciphering possible mechanisms underlying plant growth promotion and stress tolerance in rice following cyanobacterial inoculation and indicated the less explored avenue of cyanobacterial colonization in stress tolerance against abiotic stress.     

Quantitative analysis of phenolics in selected crop species and biological activity of these compounds evaluated by sensitivity of Echinochloa crus-galli

The purpose of this study was to determine the content of selected phenolic compounds in white mustard, buckwheat, spring barley, oat and rye grown under field conditions. Moreover, the allelopathic efficiency of these compounds was evaluated by sensitivity of Echinochloa crus-galli. The aromatic acids: trans-cinnamic, salicylic, ferulic, chlorogenic, p-hydroxybenzoic, protocatechuic, p-coumaric and vanillic were separated from crop plants by TLC and determined spectrophotometrically. Differences in concentrations of analysed compounds were observed for most of the examined plant species. The highest concentration was noticed for cinnamic acid and ranged from 360 μg·g⁻¹ DW in rye to 2770 μg·g⁻¹ DW in spring barley. The relatively high concentration was noticed for ferulic acid (from 73.8 μg·g⁻¹ DW in buckwheat to 1046 μg·g⁻¹ DW in spring barley) and p-coumaric acid (from 50 μg·g⁻¹ DW in oat to 1499 μg·g⁻¹ DW in buckwheat). The observed differences in the phenolics content between two successive vegetation seasons can reflect the effect of abiotic and biotic environmental factors on the phenolics level in studied plants. In the greenhouse experiment the effect of particular compounds on the growth of Echinochloa crus-galli was also studied. It has been found that the examined phenolics, and especially trans-cinnamic acid and mixture of phenolic compounds, significantly inhibit the growth of Echinochloa crus-galli. The obtained results may contribute to the explanation of the biological activity of some phenolic compounds.     

In vitro genotoxicity assessment of caffeic, cinnamic and ferulic acids

Phenols are a large and diverse class of compounds, many of which occur naturally in a variety of food plants; they exhibit a wide range of biological effects, including antibacterial, anti-inflammatory, antiallergic, hepatoprotective, antithrombotic, antiviral, anticarcinogenic, and vasodilatory actions. We examined the genotoxic and clastogenic potential of three phenolic compounds: caffeic, cinnamic and ferulic acids, using the comet and micronucleus assays in vitro. Drug-metabolizing rat hepatoma tissue cells (HTCs) were used. Three different concentrations (50, 500 and 1500 μM) of these phenolic acids were tested on the HTCs for 24 h. The caffeic, cinnamic and ferulic acids were not genotoxic by the comet assay (P > 0.05). However, the micronucleus test showed an increase in the frequency of micronucleated cells for the three compounds, indicating that these substances have clastogenic effects in HTC.     

Phenolic acid changes in mycelia of Sclerotium rolfsii as influenced by neem (Azadirachta indica) cake and Zephyarenthes citrina bulb

High performance liquid chromatographic (HPLC) analysis of mycelia of Sclerotium rolfsii grown on neem cake, and Zephyarenthes citrina bulb incorporated media was carried out. Several phenoloic acids, e.g., gallic, tannic, caffeic, cinnamic, chlorogenic and O-coumeric acids, were found in considerable amounts in treated mycelial mat as compared to the control. The amount of phenloic acids increased with increased concentration of both the materials in mycelia of 7 and 14 day-old cultures. Due to anti-oxidant and several other properties of phenolic acids, the senescence of the fungus has been prolonged which may be one probable reason of sustaining the virulence of the pathogen.     

Phenols enhancement effect of microbial consortium in pea plants restrains Sclerotinia sclerotiorum

Microbial interactions occurring in rhizosphere may play important roles in providing protection against phytopathogens. Induction of phenolic compounds and their modulation in leaf and collar region was investigated upon challenge inoculation with Sclerotinia sclerotiorum in pea plants untreated and treated with beneficial microbes viz. Bacillus subtilis BHHU100, Trichoderma harzianum TNHU27 and Pseudomonas aeruginosa PJHU15 either singly or in the form of consortium. Changes in phenolic compounds in both leaves and collar region indicated that the induced response was systemic in nature altering the physiological status of the host plant. The phenylpropanoid metabolism was strongly involved in providing resistance against S. sclerotiorum challenge; especially in the plants treated with microbes in consortium, where the response was exaggerated in order to cope up with the biotic stress induced by the pathogen. Enhanced accumulation of phenolics viz., shikimic acid, gallic acid, chlorogenic acid, syringic acid, p-coumaric acid, cinnamic acid, salicylic acid, myricitin, quercetin and kaempferol occurred both in leaf and collar tissue.     

Metabolism of phenolic compounds in <Emphasis Type="Italic">Vitis riparia</Emphasis> seeds during stratification and during germination under optimal and low temperature stress conditions

We studied the alterations in phenolic compounds in grape seeds during their stratification and germination under optimal conditions (+25 °C) and at low temperature (+10 °C). Biological materials in the study were seeds of Vitis riparia. Phenolic compounds were extracted from defatted seeds using 80 % methanol or 80 % acetone. The content of total phenolics was determined with the Folin-Ciocalteau reagent, while the content of tannins was determined by vanillin assay and the protein (BSA) precipitation method. The RP-HPLC method was used to determine phenolic compounds (phenolic acids, catechins) in the extracts. High amounts of tannins, catechins, gallic acid and lesser amounts of p-coumaric acid were found in the seeds. The content of total phenolics in acetone extracts was higher than that obtained using methanol. The amounts of phenolic acids and tannins found in V. riparia seeds after stratification were much lower. It may confirm a possible role of these compounds in dormancy of V. riparia seeds. After 72 h of low temperature treatment, inhibition of grape root growth and biochemical changes in seeds were detected. The chilling stimulated increased accumulation of some phenolic compounds (free gallic acid and catechins) in the seeds. These substances can protect plants against some abiotic stressors.