Histochemical and chemical evidence for lignin accumulation during the expression of resistance to leaf rust fungi in wheat

Levels of individual phenolic acids were examined in primary leaves of wheat (Triticum aestivum) after inoculation with avirulent and virulent strains of the leaf rust fungus (Puccinia recondita f. sp. tritici) at stages when previous work had shown fungal and host cells to be affected by expression of the Lr20 or Lr28 alleles for resistance. The predominant phenolic acid, ferulic acid, as well as p-coumaric and syringic acids were detected in primary leaves in both unbound and bound forms. They were not detected in germinating urediniospores of either rust strain. Levels of unbound phenolic acids changed little in response to infection. In Lr28-bearing leaves inoculated with an avirulent strain, increased concentrations of bound phenolic acids and three other unidentified compounds were observed about 4 h after many single or small groups of cells had undergone hypersensitive collapse. In an Lr20-bearing cultivar, levels of bound phenolic acids fell in leaves inoculated with either a virulent or avirulent rust strain. Coniferyl aldehyde and coniferyl alcohol were not detected in healthy or inoculated leaves of either wheat cultivar. Attempts to affect expression of resistance by application of inhibitors of phenylalanine ammonia-lyase were not successful and both wheat cultivars remained resistant to avirulent rust strains. The bound phenolic acids which accumulate in cells undergoing a hypersensitive response may play a role in resistance of Lr28-bearing wheat to the leaf rust fungus.     

Nature of resistance to Striga hermonthica (Del.) Benth. parasitism in some Sorghum vulgare (Pers.) cultivars

Ten Sorghum vulgare (Pers.) cultivars varying in tolerance to Striga hermonthica (Del.) Benth. parasitism were grown with or without Striga infection. Endodermal thickening, pericycle lignification and silica crystal deposition were studied microscopically and measured for infected and non-infected sorghum cultivars. Although differences in the root character measurements were statistically significant they were not closely related to the response of the plant to infection. Low stimulant producing cultivars showed low or medium root cell thickening. The cv. Framida had both low stimulant production and high root cell thickening and was the best of the tolerant cultivars. High stimulant producing, tolerant cultivars generally showed heavy or intermediate cell thickening. The high stimulant producing, susceptible cultivar Debaikri also showed intermediate root cell thickening.‘Antibiosis', measured by the content of phenolic compounds in the plant, was then studied. Varietal differences in quality and quantity of phenolic substances in the roots and shoots of sorghum cultivars infected or non-infected with Striga were observed. Infection increased total phenolic contents in both shoot and root extracts. Differences in the total phenolic content in the shoot of non-infected cultivars did not reflect tolerance to Striga infection. The total phenolic acid content of the root extracts was closely related to the response of the host plant to Striga infection, tolerant cultivars having greater total phenolic acid content than susceptible ones.     

Differential induction of redox sensitive extracellular phenolic amides in potato

This study focuses on the differential induction of extracellular phenolic amides that accumulate in potato cell suspensions during the first few hours of the interaction between these plant cells and either bacterial pathogens or pathogen-related elicitors. Using suspension cells of Solanum tuberosum we identified 4 hydroxycinnamic acid amides that accumulate in the extracellular environment. Treatment of the suspension cells with pathovars of the plant pathogens Pseudomonas syringae or Ralstonia solanacearum or with pathogen-related elicitors changed the composition of the extracellular phenolic amides within hours and the composition differed for each treatment. Some of the phenolic amides were sensitive to oxidative stress; when suspension cells were treated with bacterial strains or elicitors that triggered an oxidative burst, the phenolics were oxidized and depleted for the duration of the burst. Other critical parameters that affected the qualitative and quantitative makeup of these phenolic amides were plant cell age and density.     

Inhibition of seed germination and seedling growth by hound's-tongue (Cynoglossum officinale L.) seed leachate

The leaching of phenolics from decoated seeds, seed coats and pericarps of hound's-tongue ( Cynoglossum officinale L.), factors affecting leaching, and seed germination and seedling growth inhibitory activities of the leachate were investigated. Embryonic axes and cotyledons contained a large amount of methanol-soluble phenolic substances. Decoated seeds released phenolics into an aqueous incubation medium and this solution was capable of inhibiting root elongation of several grassy and broadleaf species, but not of hound's-tongue. The leachate and the phenolic fraction of the leachate inhibited germination of bluebunch wheatgrass ( Pseudoroegnaria spicata (Pursh) A. Löve [syn. Agropyron spicatum (Pursh) Scribn. and J. G. Sm.]). The non-phenolic fraction of the leachate did not inhibit seed germination or root and shoot elongation of bluebunch wheatgrass. Low oxygen and an increasing temperature significantly increased the amount of phenolic substances leached from decoated seeds. Increased leaching of phenolic substances related to an increasing temperature was associated with the loss of seed viability. Further investigation of the role of water-extractable hound's-tongue seed phenolics in the interaction of this species with its biotic environment under field conditions is needed to determine the ecological significance of these findings.     

Bioassays on Leptographium wingfieldii, a Bark Beetle Associated Fungus, with Phenolic Compounds of Scots Pine Phloem

The fungicidal activity of five phenolic metabolites occurring in Scots pine phloem (Cat, Tax, PS, PC and PSM) and of phenolic extracts from unwounded (fresh) and wounded phloem of Scots pine were measured against Leptographium wingfieldii, a fungus associated with Tomicus piniperda. Bioassays were performed in micro-wells in a standard medium in the absence and presence of Scots pine phloem. In both media, methanol extracts from the unwounded and wounded phloem, PS, PSM and PC inhibited fungal growth. Tax and Cat had no effect or stimulated fungal growth. The presence of Scots pine phloem in the liquid medium reduced the inhibitory effect of phenolic compounds. At low concentrations (10–4–10–5M), the fungus seemed to be able to degrade PS, PSM and PC while, whatever the Cat and Tax concentrations, it did not seem to degrade them. These results suggest that three phenolic compounds have a potential fungitoxicity in vitro and that, in vivo, these phenols could play a role in the efficiency of the induced reaction to stop fungal growth.     

Phenolic compounds as defence response of pepper fruits to Colletotrichum coccodes

Qualitative and quantitative changes of individual and total phenolics induced by Colletotrichum coccodes fungal infection have been studied in two susceptible sweet pepper cultivars ‘Soroksari’ and ‘Bagoly’, and the role of soluble phenolic compounds in plant's defence mechanism has been evaluated. Three distinct parts were analysed on pepper fruit: healthy tissue, anthracnose lesion, and bordering tissue, and individual phenolic compounds have been identified with the use of HPLC-MS system. In pepper fruit pericarp 21 phenolic compounds have been determined; the prevalent apigenin, quercetin and luteolin glycosides, chlorogenic acid and one chrysoeriol glucoside. C. coccodes infection increased the accumulation of chlorogenic acid, chrysoeriol glucoside, quercetin and luteolin glycosides in infected bordering tissue of both analysed pepper cultivars compared to healthy pepper tissue or symptomatic spot. Total apigenin derivatives did not show a significant increase in bordering tissue compared to the healthy pepper fruit in contrast to other groups of phenolics. This suggests a lesser role of apigenin glycosides in pepper plant defence against the Colletotrichum fungus. Intense phenolic synthesis was characteristic for the bordering zone between the healthy and infected plant tissue resulting in higher total phenolic content which might hinder the fungus to spread from the infected cells into the healthy tissue.     

Effect of leaf scald (<Emphasis Type="Italic">Xanthomonas albilineans</Emphasis>) on polyamine and phenolic acid metabolism of two sugarcane cultivars

Polyamine and phenolic acid levels as well as activities of some enzymes of their biosynthetic metabolism were examined in two sugarcane (Saccharum officinarum) cultivars differing in susceptibility to leaf scald, a disease caused by the bacterium, Xanthomonas albilineans. Juice obtained from both infected cultivars showed significantly increased levels of free putrescine and ornithine decarboxylase activity. However, the pathogen induced different changes in the two cultivars in subsequent metabolic steps. Whereas acid insoluble conjugated spermidine completely disappeared from the highly susceptible cv. C 439-52, an increase in acid insoluble conjugated polyamines was observed in the moderately susceptible cv. L 55-5. Phenolic acid metabolism also differed in the two cultivars. Since total phenolic acid content and phenylalanine ammonium lyase activity was greater in both cultivars after infection, distribution of phenolic acids between free or conjugated forms diverted into different pathways. The level of susceptibility of the two cultivars is discussed in terms of changes in these compounds.     

Natural phenolic acids from wheat bran inhibit <Emphasis Type="Italic">Fusarium culmorum</Emphasis> trichothecene biosynthesis <Emphasis Type="Italic">in vitro</Emphasis> by repressing <Emphasis Type="Italic">Tri</Emphasis> gene expression

The effect of natural phenolic acids from wheat bran on type B trichothecene biosynthesis by Fusarium culmorum was investigated in vitro. Durum wheat bran contained various monomeric forms of phenolic acids, with ferulic acid being the most abundant. In addition, various oligomeric forms of ferulic acid and mainly dimeric forms were also detected. When liquid cultures of F. culmorum were supplemented with a natural wheat bran extract, trichothecene production was fully inhibited. The exact mechanism by which toxin synthesis is repressed remains to be clarified but we showed that the phenolic acid treatment resulted in a drastic reduction in the expression level of structural trichothecene biosynthetic genes. The inhibitory efficiency of the natural phenolic acid extract was significantly higher than that of a reconstituted mixture containing similar concentrations of monomeric forms. Thus, to elucidate the full repression of type B trichothecene production induced by the natural phenolic acid extract from wheat bran, two hypotheses can be raised: (i) a synergistic impact of monomeric and dimeric forms of phenolic acids, (ii) the occurrence of an unidentified oligomeric form able to efficiently repress toxin yield. As a first attempt to investigate the effect of oligomeric forms, one of the most abundant dimer of ferulic acid, the 8-5′-benzofuran dimer, has been synthesized in vitro and was shown to inhibit trichothecene biosynthesis to the same extent than the monomer of ferulic acid.     

Dynamic root exudation of phenolic allelochemicals from Johnson grass (Sorghum halepense)

The phenolic allelochemicals from the root exudates of Johnson grass (Sorghum halepense) were collected at different developmental stages and analyzed using high performance liquid chromatography‐ultraviolet/photo‐diode array. The results revealed that three simple phenolic components, p‐hydroxybenzoic acid, p‐hydroxybenzaldehyde and ethyl p‐hydroxybenzoate, along with two flavonoids, diosmetin (3′,5,7‐trihydroxy‐4′‐methoxyflavone) and tricin (5,7,4′‐trihydroxy‐3′,5′‐dimethoxyflavone), were found in the root exudates of Johnson grass. However, the major phenolic compounds varied and the quality of each compound was dynamic at different developmental stages: the only phenolic compound that was detected in the seedling stage was p‐hydroxybenzoic acid; three simple phenolic allelochemicals, p‐hydroxybenzoic acid, p‐hydroxybenzaldehyde and ethyl p‐hydroxybenzoate, were found in the jointing stage; and five compounds, including the two flavonoids, tricin and diosmetin, appeared in the reproductive stage along with the other three simple phenolic allelochemicals. The highest diversity in phenolic constituents was in the reproductive stage, with five allelochemicals. Only p‐hydroxybenzoic acid was found in the whole life cycle of Johnson grass, with a range of 2.9 nmol to 6.8 nmol per plant per day. p‐Hydroxybenzaldehyde had the highest quality among all the compounds, with the highest releasing rate of 26.8 nmol per plant per day. The lowest‐content compounds were the two flavonoids, with the highest quality of 1.8 nmol per plant per day. The results suggest that the temporal dynamic of phenolic allelochemicals in the root exudates of Johnson grass corresponds with the dynamic of development.     

Interaction Between Nitrogen-Fertilized Peach Trees and Expression of syrB, a Gene Involved in Syringomycin Production in Pseudomonas syringae pv. syringae

ABSTRACT The in vitro expression of the syrB gene that controls the synthesis of syringomycin, a non-host-specific phytotoxin produced by Pseudomonas syringae pv. syringae van Hall, was studied using aqueous extracts derived from bark tissues collected from nitrogen-fertilized and nonfertilized peach trees. Expression of the syrB gene was quantified as beta- galactosidase activity expressed by P. syringae pv. syringae B3AR-132 containing a syrB::lacZ fusion. Gene expression was significantly less in three of four paired comparisons using extracts derived from fertilized versus nonfertilized trees; however, canker lengths were significantly different in only one of four comparisons. Expression was negatively correlated with plant tissue nitrogen content and positively correlated with a plant carbon/nitrogen ratio. Bark tissue from ring nematodeinfested trees had significantly higher concentrations of total soluble phenolic compounds and carbon/nitrogen ratios than bark samples from trees without nematodes, and canker size was significantly greater in trees growing in ring nematode-infested soil compared with noninfested soil. Nitrogen fertilization significantly decreased the plant carbon/nitrogen ratio, which was positively correlated with the concentration of total soluble phenolic compounds. Canker size developing after bacterial inoculation was positively correlated with higher plant carbon/nitrogen ratios and total soluble phenolic compounds. These results support the hypothesis that one reason why nitrogen fertilization decreases host susceptibility to bacterial canker is by either reducing the amount of plant metabolites that can induce syrB gene expression, or producing or increasing the concentration of compounds that antagonize syrB inducing compounds.     

三种酚酸类化合物对莴苣幼苗的化感作用及机理初探

化感物质通过植物体释放到农田土壤中,可对作物生长造成严重影响。酚酸是一类重要的化感物质,由于其相对分子质量小,结构稳定,因而不易降解而累积在土壤中,进而影响作物生长,是引起连作障碍问题的一类主要化合物。为了明确酚酸类物质对植物生长影响的途径,探讨其化感作用机理,以3种结构相似的化合物——苯甲酸、水杨酸和对羟基苯甲酸为研究对象,以莴苣幼苗为活性受体,通过测定化合物对莴苣幼苗根、茎生长的影响,评价了其对植物的毒活性;并借助于荧光显微镜,通过FDA/PI双染色法和DHE染色法,分别研究了3种酚酸类化合物对受体植物根尖细胞活力和活性氧积累的影响。结果表明:3种酚酸类化合物对莴苣幼苗根、茎的生长均表现出低浓度促进、高浓度抑制的作用模式;莴苣幼苗在高浓度酚酸的胁迫下,其体内活性氧大量积累,高浓度的活性氧对莴苣幼苗细胞膜结构等具有显著的破坏作用,进而导致细胞活力降低甚至凋亡,最终抑制植物生长。     

Allelopathic effect of the root exudates of K21, a potent allelopathic rice

An inhibitory effect of the phenolic compounds released from the roots of K21, a newly bred potent allelopathic rice, was investigated in comparison to its male parent, Kouketsumochi, and its female parent, Dongjinbyeo. The inhibitory rates of the root exudates of K21 and both of its parents, collected at 5 day intervals, on the shoot growth of Echinochloa crus-galli var. praticola exhibited an increasing trend from 19–34 days after seeding (DAS). However, the root exudates of K21, at concentrations of 0.5, 1.0, and 2.0%, showed a greater inhibitory effect than those of its female parent on the growth of E. crus-galli var. praticola at the earlier collection dates. Nine phenolic compounds were identified; the total amount of phenolic compounds in the exudates increased from 19–34 DAS in both K21 and its male parent, but not much in the female parent. The subsequent increase in the amount of total phenolic compounds in the root exudates of the later investigation dates and the trend of increasing inhibitory rates on the shoot growth of E. crus-galli var. praticola at higher exudate concentrations imply that the increased amount of phenolic acids in the root exudates of K21 secreted on the subsequent dates could be related to its greater allelopathic effects.     

Testing the systemic induced resistance hypothesis with Austrian pine and Diplodia sapinea

Systemic induction of defenses (e.g. phenolic metabolites) is considered vital in conifer resistance to pathogens and insects, and forms the mechanistic basis of the systemic induced resistance hypothesis (SIRH). In this study, the SIRH was tested on juvenile Austrian pine. Main stems expressed SIR in a manner that was consistent with the SIRH, while shoots became uniformly more susceptible to subsequent inoculations, demonstrating clear organ specificity in the tree's response. The majority of phenolic metabolites were poorly correlated with phenotype. Thus, the defensive system of Austrian pine is highly plastic and organ specific, and cannot be predicted by phenolic profiles alone.     

Epidemiological and histological components of crown rust resistance in oat genotypes

Crown rust, caused by Puccinia coronata f. sp. avenae, can cause significant damage in all regions where oats (Avena sativa L.) are cultivated. The primary means of controlling crown rust has been through genetic resistance, although in most cases resistance has been quickly overcome by the pathogen. More durable partial or non-specific resistance may possess different mechanisms from those underlying genes with specific effects. We studied the epidemiological and histological components of crown rust resistance with potential use in plant protection. Among the components evaluated, pustule density showed the clearest effect on resistance, while the latent period was not an important component. Cell death associated with the accumulation of autofluorescent and phenolic compounds was common in the resistant genotypes, but temporally distinct for the genotypes studied. Genotype Pc68/5*Starter, which has race-specific resistance, showed rapid cell death that prevented the development of pathogen colonies. Conversely, with cultivar URS 21 and genotypes 04B7113-1 and 04B7119-2, cell death and associated accumulation of autofluorescent and phenolic compounds was delayed until pathogen colonies were already established. Pathogen colonies developed normally in susceptible plants genotypes, and had usually produced sporogenic tissue by 5 days after inoculation. The data suggest that the resistance mechanisms, especially hypersensitivity and phenolic compound production, active in resistant plants are similar but may be differently expressed over time. The temporal variation in the expression of hypersensitivity and phenolic compound production reflects the level of field resistance in these genotypes.     

Grapevine phenolic compounds in xylem sap and tissues are significantly altered during infection by Xylella fastidiosa

Pierce's disease of grapevine (PD), caused by the bacterial pathogen Xylella fastidiosa, remains a serious problem for grape production in California and elsewhere. This research examined induction of phenolic compounds in grapevines ('Thompson Seedless') infected with X. fastidiosa over a 6-month period. Two months postinoculation with X. fastidiosa, catechin, digalloylquinic acid, and astringin were found at greater levels in xylem sap; multiple catechins, procyanidins, and stilbenoids were found at greater levels in xylem tissues; and precursors to lignin and condensed tannins were found at greater levels in xylem cell walls. However, such large-scale inductions of phenolic compounds were not observed 4 months after inoculation. Six months after inoculation, infected plants had significantly reduced phenolic levels in xylem sap and tissues when compared with control plants, including lowered levels of lignin and condensed tannins. At 6 months, PD symptoms were severe in infected plants and most photosynthetic tissue was abscised. These results suggest that, even though grapevine hosts may initially respond to X. fastidiosa infections with increased production of phenolic compounds, ultimately, PD causes grapevines to enter a state of decline whereby diseased hosts no longer have the resources to support secondary metabolite production, including defense-associated phenolic compounds.     

Antifungal capacity of major phenolic compounds of Olea europaea L. against Phytophthora megasperma Drechsler and Cylindrocarpon destructans (Zinssm.) Scholten

The phenolic composition of olive roots and stems was studied by high performance liquid chromatography–mass spectrometry. The in vivo levels of the principal phenolic compounds found in olive plants infected by Phytophthora megasperma Drechsler and Cylindrocarpon destructans (Zinssm.) Scholten differed from the levels observed in non-infected plants. When the antifungal activity of these compounds against both fungi was studied in vitro, the most active were quercetin and luteolin aglycons, followed by rutin, oleuropein, p-coumaric acid, luteolin-7-glucoside, tyrosol and catechin. Microscopic study showed that these phenolic compounds affected the growth, morphology and ultrastructure of the fungi. Taken together, these findings suggest that the phenolic compounds present in olive plants play an active role in the protection against pathogen attack.     

Differential Inhibition of Sphaeropsis sapinea Morphotypes by a Phenolic Compound and Several Monoterpenes of Red Pine

ABSTRACT The in vitro effects of a red pine phenolic compound (pinosylvin), a phenolic compound common to other species (tannic acid), and the major red pine monoterpenes (alpha-pinene, beta-pinene, and delta-3-carene) on spore germination and mycelial growth of Sphaeropsis sapinea were examined. Two A and two B morphotype isolates were used. At 88 mug/mm(2), pinosylvin inhibited spore germination of all four isolates (98 to 100%). At 8.8mug/mm(2), spore germination of B isolates was inhibited more than that of A isolates (73 versus 30%). Pinosylvin also inhibited mycelial growth of B isolates more than that of A isolates (84 versus 13% at 88 mug/mm(2)). Tannic acid stimulated or had little affect on spore germination and had little affect on mycelial growth of either morphotype. Spore germination of B isolates was inhibited more than that of A isolates by beta-pinene at saturation (79 versus 37%). Spore germination of B isolates was inhibited and germination of A isolates was stimulated by delta-3-carene below saturation (49 versus -7%). Mycelial growth of B isolates was inhibited more than that of A isolates by all monoterpenes at saturation. Differences observed between morphotypes below saturation were significant only for beta-pinene. These results demonstrate the biological activity of a phenolic compound and monoterpenes that occur in red pine. The differential responses might provide means of distinguishing morphotypes and offer a potential explanation for ecological specialization.     

Soluble phenolic acids in Capsicum annuum stems infected with Phytophthora capsici

The soluble phenolic acids of the stems of three Capsicum annuum cultivars associated with differing resistance to Phytophthora capsici showed qualitative and quantitative variation after inoculation, notably an increase in the total phenolic acid content in the resistant and intermediate varieties. The soluble phenolic acids also retarded myeelial growth of P. capsici in culture. The most pronounced inhibitory effect was produced by f-cinnamic acid, followed by p-hydroxybenzoic, vanillic and salicylic acids. Colonization and necrosis spread throughout the stem of the susceptible cultivar whereas the stem of the resistant cultivar was only partially invaded. In the latter, invasion of the tissue adjacent to necrosis occurred in only 20% of the plants.     

Comparative effects of dichlobenil and its phenolic alteration products on photo- and oxidative phosphorylation

Effects of dichlobenil (2,6-dichlorobenzonitrile) and its phenolic degradation products (2,6-dichloro-3-hydroxybenzonitrile and 2,6-dichloro-4-hydroxybenzonitrile) were compared on electron transport and phosphorylation in isolated spinach (Spinacia oleracea L.) chloroplasts and mung bean (Phaseolus aureus Roxb.) mitochondria. In chloroplasts, the hydroxylated derivatives inhibited both photoreduction and coupled photophosphorylation with water as the electron donor and with ferricyanide as oxidant, and cyclic photophosphorylation with phenazine methosulfate as the electron mediator under an argon gas phase. In mitochondria, the phenolic derivatives acted as uncouplers of oxidative phosphorylation as evidenced by the stimulation of ADP-limited respiration, circumvention of oligomycin-inhibited non-ADP-limited respiration, and the induction of ATPase activity. Treatment of excised mung bean hypocotyls by the phenolic derivatives also resulted in a very rapid and drastic lowering of ATP levels. In all assays, only limited, if any, interference was expressed by dichlobenil even at relatively high molar concentrations.Inhibition of oxidative and photophosphorylation by the phenolic degradation products, but not by dichlobenil, suggests that if there is a delay between the formation of the hydroxylated compounds and their conjugation, photosynthesis and respiration will be inhibited. Because biochemical and physiological processes depend on oxidative and photophosphorylation for the energy (ATP) needed to drive the reactions, interference with ATP production could be one of the major mechanisms through which phytotoxicity is expressed by the phenolic degradation compounds of the herbicide, if they should accumulate in the free from. Species selectivity may be related to the rate of formation of the phenolic products in different plants and the rapidity of conjugate formation.