A simple method for visualization of phenolics exudation by roots of intact lupin plants; the effect of nitrate and pH.

Phenolics exudation by imbibed seeds and roots of intact lupin plants (Lupinus albus L.) was studied during the first 4 days of growth by a new agar test with specific reagents for phenolics (Gibbs reagent, Naturstoffreagenz A). Comparative studies of the phenolics exudation reveal that legumes exude different phenolics (even if not qualitatively, then at least quantitatively) than oat. The exudation of phenolics starts very quickly after the imbibition of seeds and can be visualized as early as 24 h after sowing. In older seedlings, the exudation of phenolics can be detected along root zones and is influenced by nitrate and pH. At acidic pH, nitrate reduces phenolics exudation, but at pH 7.5 the exudation of phenolics becomes restricted to only some root zones. Nitrate must be present in the rooting media for at least 24 h to cause visible changes in the pattern of exudation at different pH values.     

Effective Phytoextraction of Cadmium (Cd) with Increasing Concentration of Total Phenolics and Free Proline in Cannabis sativa (L) Plant Under Various Treatments of Fertilizers,Plant Growth Regulators and Sodium Salt

The comparative effect of fertilizers (NPK), plant growth regulators (GA₃, IAA, Zeatin) and sodium chloride (NaCl) on Cd phytoaccumulation, proline and phenolics production in Cannabis sativa was evaluated. Proline and phenolices were correlated with Cd contents in plant. Cd significantly reduced the plant growth. Fertilizers application (in combination) most significantly increased the growth (19 cm root and 47 cm shoot) on Cd contaminated soil. All treatments increased the Cd contents in plant tissues. This increase was highly significant in fertilizers treated plants (1101, 121 and 544 ppm in roots, stem and leaves respectively). Significantly positive correlation was found between Cd concentration and dry biomass of root (R² = 0.7511) and leaves (R² = 0.5524). All treatments significantly increased the proline and total phenolics and maximum was recorded in NaCl treated plants followed by fertilizers. Proline was higher in roots while phenolics in leaves. The correlation between proline and phenolics was positive in leaf (R² = 0.8439) and root (R² = 0.5191). Proline and phenolics showed positive correlation with Cd concentration in plant. Conclusively, fertilizers in combination seem to be the better option for Cd phytoextraction. Further investigation is suggested to study the role of phenolics and proline in Cd phytoextraction.     

The Iron-Deficiency Induced Phenolics Accumulation May Involve in Regulation of Fe(III) Chelate Reductase in Red Clover

Although considerable researches have been conducted on the physiological responses to plant iron (Fe) deficiency stress in dicotyledonous plants, much still needs to be learned about the regulation of these processes. In the present research, red clover was used to investigate the role of root phenolics accumulation in regulating Fe-deficiency induced Fe(III) chelate reductase (FCR). The root FCR activity, IAA and phenolics accumulation, and also the phenolics secretion were greatly increased by the Fe deficiency treatment. The application of TIBA (2,3,5-triiodobenoic acid) to the stem, an IAA polar transport inhibitor, which could decrease IAA accumulation in root, significantly inhibited the FCR activity, but did not effect root phenolics accumulation and secretion, suggesting that IAA itself did not involve in root phenolics accumulation and secretion. In contrast, the Fe deficiency treatment significantly decreased the root IAA-oxidase activity. Interestingly the phenolics extracted from roots inhibited IAA-oxidase activity in vitro, and this inhibition was greater with phenolics extracted from roots of Fe deficient plants than that from Fe sufficient plants, indicating that the Fe deficiency-induced IAA-oxidase inhibition probably caused by the phenolics accumulation in Fe deficient roots. Based on these observations, we propose a model where under Fe deficiency stress in dicots, an increase in root phenolics concentrations plays a role in regulating root IAA levels through an inhibition of root IAA oxidase activity. This response, leads to, or at least partially leads to an increase in root IAA levels, which in turn help induce increased root FCR activity.Key Words: Fe deficiency, ferric chelate reductase, phenolics, Trifolium pretense     

What causes changes in plant litter quality and quantity as consequence of grazing in the Patagonian Monte: Plant cover reduction or changes in species composition?

In Patagonian Monte, as in other arid ecosystems, grazing has triggered changes in vegetation and soil such as plant cover reduction, changes in species composition and soil nutrient losses. Several mechanisms were proposed interconnecting these changes, but evidence supporting them is very scarce. On the basis of published data concerning plant cover by species along grazing gradients and leaf litter production of dominant species, we estimated the effects of grazing on a – quality (N, soluble phenolics and lignin concentrations) and b – quantity (leaf litterfall (LLF) and inputs of nitrogen, soluble phenolics and lignin to the soil) of leaf litter in the Patagonian Monte, discriminating the effect of plant cover reduction from that of species composition. We also evaluated the relationship between senesced leaves traits and the response of species to grazing (i.e. their relative change in plant cover). Grazing causes a reduction in LLF and in the inputs of nitrogen, soluble phenolics and lignin to the soil. In the case of LLF, this reduction was not only a result of the decrease in plant cover but also due to changes in species composition. In contrast, our results showed that the reduction in nitrogen, soluble phenolics and lignin inputs to the soil by LLF is only a consequence of plant cover reduction. Additionally, litter quality was affected through increasing concentration of N and secondary compounds (soluble phenolics and lignin). N and soluble phenolics concentration on senesced leaves were positively related to the response of species to grazing, suggesting that other factors instead of N are relevant to sheep foraging decisions.     

UV radiation is the primary factor driving the variation in leaf phenolics across Chinese grasslands

Due to the role leaf phenolics in defending against ultraviolet B (UVB) under previously controlled conditions, we hypothesize that ultraviolet radiation (UVR) could be a primary factor driving the variation in leaf phenolics in plants over a large geographic scale. We measured leaf total phenolics, ultraviolet‐absorbing compounds (UVAC), and corresponding leaf N, P, and specific leaf area (SLA) in 151 common species. These species were from 84 sites across the Tibetan Plateau and Inner Mongolian grasslands of China with contrasting UVR (354 vs. 161 mW/cm² on average). Overall, leaf phenolics and UVAC were all significantly higher on the Tibetan Plateau than in the Inner Mongolian grasslands, independent of phylogenetic relationships between species. Regression analyses showed that the variation in leaf phenolics was strongly affected by climatic factors, particularly UVR, and soil attributes across all sites. Structural equation modeling (SEM) identified the primary role of UVR in determining leaf phenolic concentrations, after accounting for colinearities with altitude, climatic, and edaphic factors. In addition, phenolics correlated positively with UVAC and SLA, and negatively with leaf N and N: P. These relationships were steeper in the lower‐elevation Inner Mongolian than on the Tibetan Plateau grasslands. Our data support that the variation in leaf phenolics is controlled mainly by UV radiation, implying high leaf phenolics facilitates the adaptation of plants to strong irradiation via its UV‐screening and/or antioxidation functions, particularly on the Tibetan Plateau. Importantly, our results also suggest that leaf phenolics may influence on vegetation attributes and indirectly affect ecosystem processes by covarying with leaf functional traits.     

Interactive effects of leaf maturation and phenolics on consumption and growth of a geometrid moth

Phenolic compounds are commonly regarded as the main chemical defenses of deciduous woody plants against insects. To examine how indices of leaf maturation (water content, toughness, and sugar/protein ratio) modified larval consumption and growth relative to phenolics and phenolic-related leaf traits, we measured consumption and growth of fourth-instar Epirrita autumnata (Bkh.) (Lepidoptera: Geometridae) larvae on three different days on young, normal, and mature leaves, respectively, from the same mountain birch (Betula pubescens ssp. czerepanovii (Orlova) Hämet-Ahti) trees. The larvae achieved the same growth rates on young and normal leaves, but had to consume 40% more on the latter. On more mature leaves, larval growth was poorer and was positively correlated with sugar/protein ratios (although the ratio peaked at that time). Indices of leaf maturation correlated with several phenolics in data pooled over the three study days, but poorly in any individual day. Similarly, in the pooled data, larval consumption and growth correlated with several leaf traits, but correlations between leaf and insect traits were few on any of the three days, and no trait was significant on each of the three days.We next examined whether variation in the maturation indices modified the associations of phenolics with insect consumption and growth. When interactions between phenolics and leaf maturation indices were taken into account, the number of phenolic compounds displaying significant associations with insect traits more than doubled. The relative importance of interactive versus direct associations increased with leaf maturation: on young leaves five phenolics showed direct and eleven interactive associations with insect traits, while in mature leaves we found two phenolics to display direct and thirteen phenolics interactive associations. Leaf water content, either alone or together with toughness and sugar/protein ratio, generally explained more of the variance in Epirrita growth (up to 59%) than any phenolic or phenolic-related trait alone (highest value 20%). Including interactive effects between phenolics and indices of leaf maturation in the model increased the proportion explained of variance in larval growth between 49 and 73%. Maturation indices explained 0 to 23% of variance in consumption, and the phenolic compound with the highest (positive!) correlation alone up to 28%, but taking into account interactions between phenolics and maturation indices raised the degree of explanation much (namely, 32 to 53%) over that explained by indices of leaf maturation alone. This indicates strong interactive effects on consumption between phenolics and indices of leaf maturation.     

Detoxification of sugarcane bagasse hydrolysate improves ethanol production by Candida shehatae NCIM 3501

Sugarcane bagasse hydrolysis with 2.5% (v/v) HCl yielded 30.29g/L total reducing sugars along with various fermentation inhibitors such as furans, phenolics and acetic acid. The acid hydrolysate when treated with anion exchange resin brought about maximum reduction in furans (63.4%) and total phenolics (75.8%). Treatment of hydrolysate with activated charcoal caused 38.7% and 57.5% reduction in furans and total phenolics, respectively. Laccase reduced total phenolics (77.5%) without affecting furans and acetic acid content in the hydrolysate. Fermentation of these hydrolysates with Candida shehatae NCIM 3501 showed maximum ethanol yield (0.48g/g) from ion exchange treated hydrolysate, followed by activated charcoal (0.42g/g), laccase (0.37g/g), overliming (0.30g/g) and neutralized hydrolysate (0.22g/g).     

Randomized controlled trial of dietary intervention: association between level of urinary phenolics and anti-mutagenicity

We have undertaken a randomized trial to confirm the ability of a class of phenolics, flavonoids, to increase urinary anti-mutagenicity in smokers. Ninety heavy smokers were recruited and randomly assigned to three groups, who were given three different diets. One diet was rich in flavonoids, but not based on supplementation ('flavonoid'), one was a normal iso-caloric diet with an adequate administration of fruit and vegetables ('normal'), and one was based on supplementation of flavonoids in the form of green tea and soy products ('supplement'). The urinary anti-mutagenicity-as inhibiting effect of the urinary extracts on the mutations induced by MeIQx-was measured in Salmonella typhimurium YG1024 in the presence of liver S9 from male Sprague-Dawley rats treated with Aroclor 1254. The amount of total phenolics in the urinary extracts was measured by use of spectrometric analysis. We found that important dietary modifications can be achieved through special recipes and instructions given by a cook during an intensive course. The intervention was focused on increasing the flavonoid intake, and it was successful in that respect. In fact, differences in flavonoid intake were appreciated mainly between the first group (normal diet) and the other two (flavonoid-rich and supplemented diet), suggesting that dietary modification can be as effective as supplementation. However, both urinary anti-mutagenicity and the amounts of urinary phenolics did not change as a consequence of the trial. These results suggest that only a small fraction of urinary phenolics is influenced by dietary changes in the intake of flavonoids, and that most urinary anti-mutagens and phenolics are metabolites of dietary flavonoids, whose formation is more affected by the activity and diversity of bacterial flora in the colon than by the quantity and type of intake. A strong correlation was found between urinary phenolics and anti-mutagenicity in all the groups involved in the trial. Such correlation was not explained by dietary variables.     

Production of phenolic antioxidants by the solid-state bioconversion of pineapple waste mixed with soy flour using Rhizopus oligosporus

The ability of Rhizopus oligosporus to produce enhanced levels of free phenolics from pineapple residue mixed with soy flour as potential nitrogen source was investigated. Concurrently, phenolic-linked β-glucosidase and the antioxidant activity of the extracts were followed. Two treatments were studied: (A) 9 g of pineapple residue and 1 g of soy flour (P9); (B) 5 g of pineapple residue and 5 g of soy flour (P5). The increase of water extractable phenolics was 39.3% for P9 treatment and 79.4% for P5 treatment. During early stages of growth high antioxidant activity, low phenolic content and low β-glucosidase specific activity was observed. High antioxidant activity was likely due to the presence of insoluble polymeric phenolics, know for their high antioxidant activity. A marked decrease of the antioxidant activity of P5 treatment during late stages of growth was observed due to likely formation of free soluble phenolics. The moderate total phenolics content and high β-glucosidase specific activity of P9 treatment in late stages is likely the consequence of low nitrogen content in this treatment. The bioconversion of pineapple residue by R. oligosporus leads to enhanced levels of phenolic compounds, mainly for P5 treatment. This approach offers a novel strategy to enhance the value of pineapple wastes.     

A rice phenolic efflux transporter is essential for solubilizing precipitated apoplasmic iron in the plant stele

Iron deficiency is one of the major agricultural problems, as 30% of the arable land of the world is too alkaline for optimal crop production, rendering plants short of available iron despite its abundance. To take up apoplasmic precipitated iron, plants secrete phenolics such as protocatechuic acid (PCA) and caffeic acid. The molecular pathways and genes of iron uptake strategies are already characterized, whereas the molecular mechanisms of phenolics synthesis and secretion have not been clarified, and no phenolics efflux transporters have been identified in plants yet. Here we describe the identification of a phenolics efflux transporter in rice. We identified a cadmium-accumulating rice mutant in which the amount of PCA and caffeic acid in the xylem sap was dramatically reduced and hence named it phenolics efflux zero 1 (pez1). PEZ1 localized to the plasma membrane and transported PCA when expressed in Xenopus laevis oocytes. PEZ1 localized mainly in the stele of roots. In the roots of pez1, precipitated apoplasmic iron increased. The growth of PEZ1 overexpression lines was severely restricted, and these lines accumulated more iron as a result of the high solubilization of precipitated apoplasmic iron in the stele. We show that PEZ1 is responsible for an increase of PCA concentration in the xylem sap and is essential for the utilization of apoplasmic precipitated iron in the stele.     

四川王朗自然保护区缺苞箭竹（Fargesia denudate Yi.）总酚含量及变化规律

采用分光光度法对四川王朗自然保护区缺苞箭竹的总酚含量进行了分析,结果表明:在采集到的不同器官中均有酚类物质分布,总酚含量最高的为叶龄超过1a的老叶(2.05%),笋中的总酚含量最低(0.20%).海拔高度和日照强度在一定程度上影响其含量变化,在海拔高和光照强的条件下,幼嫩器官中的总酚含量有明显增加的趋势.研究分析了缺苞箭竹各器官总酚含量的变化规律,及其与环境因子和大熊猫采食量之间的关系.     

Antiviral and virucidal activities against herpes simplex viruses of umesu phenolics extracted from Japanese apricot

Umesu phenolics were obtained from the salt extracts of Japanese apricot (Nanko‐mume cultivar of Prunus mume Sieb. et Zucc.) as purified phenolics. The antiviral activities of umesu phenolics obtained were then examined against herpes simplex virus type 1 (HSV‐1) and type 2 (HSV‐2), enveloped DNA viruses. The phenolics inhibited the multiplication of these viruses when added to the culture media of the infected cells. This inhibition occurred at phenolic concentrations at which they showed no severe cytotoxicity. One‐step growth experiments showed that the eclipse period in the HSV‐1 multiplication process was extended in the presence of umesu phenolics and that the addition of phenolics after the completion of viral DNA replication did not affect their multiplication. More drastic effects were observed on virucidal activities against HSV‐1 and HSV‐2; the infectivity decreased to 0.0001 when infected cells were incubated with 3 mg/ml phenolics at 30°C for 5 min. These results demonstrate the antiviral and virucidal activities of umesu phenolics and suggest a potential pharmacological use for these phenolics as a sanitizing or preventive medicine against superficial HSV infections.     

Temporary immersion bioreactors (TIB) provide a versatile,cost-effective and reproducible in vitro analysis of the response of pineapple shoots to salinity and drought

Effects of salinity (NaCl) and the carbon source mannitol (0–200 mM) on micropropagation of pineapple cv. MD2 were analyzed in temporary immersion bioreactors (TIBs). Shoot multiplication rate, shoot cluster fresh weight and levels of aldehydes, chlorophylls, carotenoids and phenolics were determined in the plant material. The content of soluble phenolics in the culture medium was also evaluated. NaCl or mannitol above concentrations of 50 mM decreased pineapple shoot multiplication and fresh weight significantly. Two hundred mM NaCl decreased multiplication rate by 71.5% and cluster fresh weight by 40.0%. NaCl increased 2.4 times the levels of other aldehydes; 1.4 times the soluble phenolics in shoots; and 1.4 times the phenolics excreted to the culture medium. On the other hand, mannitol decreased the multiplication rate and cluster fresh weight by about 60%. Mannitol increased the contents of chlorophyll b 1.4 times and soluble phenolics 2.1 times. Results indicated that pineapple cv. MD2 is more sensitive to NaCl than to mannitol. Multiplication rates indicate that a 50% reduction was obtained with 37.4 mM NaCl and 66.5 mM mannitol. These concentrations can be used to stress shoots during micropropagation in TIBs and screen for/detect somaclonal variants with an increased salinity or drought tolerance.     

Effect of nitrogen fertilization on leaf phenolic production of grand fir seedlings

Summary A significant decrease in production of total phenolics was apparent with nitrogen fertilization in three of five seed sources of grand fir [Abies grandis (Dougl.) Lindl.]. Subsequent extraction and identification of phenolics indicated that two compounds increased with fertilization, and when grouped by biosynthetic origin into cinnamic acids, benzoic acids, and flavonoids, no differences were apparent with fertilization. Total biomass increased with fertilization, and the relation between growth and total phenolics was similar for all seed sources. The increase in growth and decrease in total phenolics suggest that, in the carbon allocation hierarchy, as available soil nitrogen increases, phenolics represent a lower priority.     

Antioxidative activity of bound-form phenolics in potato peel

Free and bound-form phenolics were isolated from potato (cv. Toyoshiro) flesh and peel. The free and bound-form phenolics in the peel showed high DPPH radical scavenging activity, while those in the flesh showed low activity. The total amount of chlorogenic acid and caffeic acid in the free-form phenolics from the peel was highly correlated with the DPPH radical scavenging activity. Ferulic acid was identified as the active radical scavenging compound in the bound-form phenolics from the peel. The potato peel may therefore offer an effective source of an antioxidative.     

Changes in the concentration of phenolic compounds and exudation induced by phosphate deficiency in bean plants (Phaseolus vulgaris L.)

The effect of prolonged phosphate starvation of bean plants (Phaseolus vulgaris L.) on the concentration of phenolics and their exudation by roots was studied. Plants cultured on phosphate-deficient media maintained a steady concentration of total phenolics in the leaves, whereas in the leaves of plants grown on complete nutrient media the phenolic concentration decreased. After 18 days of culture, higher total phenolics and anthocyanin concentrations in phosphate-deficient leaves compared with control leaves were observed. The divergent trends in total phenolic concentrations between phosphate-deficient and control leaves corresponded to the changes in the activity of L-phenylalanine ammonia-lyase. In the roots, the concentration of total phenolics was lower in phosphate-deficient plants compared with control plants. However, after 18 days of culture of bean plants, the amount of exuded phenolics from phosphate-deficient roots was 5-times higher than that from the roots of control plants. The activity of L-phenylalanine ammonia-lyase was twice as high in the roots of phosphate-starved plants. Comparable rates in the exudation of phenolics by bean roots observed after 18 days of culture on nitrogen-deficient or phosphate-deficient medium may suggest a similar system of signal transduction for phenolics release. The results are discussed in relation to the possible functions of phenolics in nutrient uptake and as chemical signals in root-soil microbe interactions to enhance the plant adaptation to particular environmental conditions.     

Radical scavenging potential of phenolics from Bryophyllum pinnatum (LAM.) OKEN

Optimization of the extraction process of phenolics from Bryophyllum pinnatum was carried out using response-surface methodology (RSM). The effect of different variables such as ratio of solvents, plant material/solvent ratio, extraction time, and temperature were investigated. An optimal phenolics yield of 7.952 mg/g gallic acid equivalence (GAE) was achieved at reduced levels of methanol/water ratio (1:1, v/v). During optimization, the product yield was enhanced by ～2-fold at reduced extraction solvent (methanol/water) up to 37%. Validation of the RSM model for extraction of total phenolic content (TPC) was confirmed by high-performance liquid chromatography (HPLC) analysis. The obtained experimental values were in good agreement with the predicted values, thereby indicating the appropriateness of the model generated. Phenolic extracts from B. pinnatum were further examined by 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) methods for determining the radical scavenging activities. EC(50) values of B. pinnatum extracts (BPEs) obtained by these methods were in accordance with the amount of phenolics present in the extract. Significant correlation was found between total phenolic content and antioxidant activities (p < 0.05).     

Phenolic content and growth of wetland macrophytes: Is the allocation to secondary compounds driven by nutrient availability?

This study compares soluble phenolics and lignin content in two wetland macrophytes with contrasting life strategies grown under a varying nutrient supply in the field and in a greenhouse experiment. The differences are explained in terms of the protein competition model (PCM) hypothesis relating changes in secondary metabolites to changing nutrient limitation. The two study species, Eleocharis cellulosa (EC) and Typha domingensis (TD), are both widespread in tropical and subtropical freshwater and brackish marshes of the New World, and are often found in P-limited rather than N-limited conditions. TD is a fast-growing competitor with large nutrient requirements. EC is a stress tolerator, quite well adapted to growth in nutrient-limiting environments. In both species, the concentration of phenolics was negatively correlated with increasing growth (due to increasing nutrient levels). This is in agreement with the PCM hypothesis, which predicts an increase in phenolic synthesis when protein synthesis (and consequently growth) is low due to limited resource availability. An interesting difference was found in the correlation between tissue nutrients and phenolics. TD from both the field and the greenhouse showed a negative correlation between tissue P and phenolics, while EC displayed a significant negative correlation between tissue N and phenolics. EC is adapted to low P, and increased tissue P content represents luxury consumption (uptake of P for storage) which is not reflected in increased growth and thus is not correlated with phenolics. These are the first steps in elucidating the relationship among nutrient availability, growth and phenolic content in two important primary producers of tropical and subtropical marshes.     

Biochemical Aspects of Parasitism by the Angiosperm Parasites: Phenolics in Parasites and Hosts

the contents of total phenolics in three parasitic angiosperms, Cuscuta species, Orobanche aegyptiaca and Dendrophthoe falcata and their respective hosts, were colorimetrieally determined. A biochemical comparison was made of the phenolics on the basis of the ability of alcoholic extracts of the tissues to inhibit amylose phosphorylase in vitro. High concentration of phenolics seemed to be a general feature of parasitic angiosperms. An increase in the concentration of the phenolics occurred in the tissues of infected hosts, in comparison with controls. the phenolics of Orobanche and mistletoe had inhibitory activity against amylose phosphorylase, but those of Cuscuta developed the inhibitory ability only when growing on hosts which themselves possessed inhibitory phenolics. the inhibitory activity of host phenolics was sometimes altered as a result of infection by parasite. Although the hosts often exerted some influence on the concentration and the inhibitory activity of phenolics in the parasites, there was no direct relationship between host and parasite phenolics. the sum of the phenolics in the tissues of parasite and the infected bost generally exceeded the phenolics in the tissues of the control host. The content of phenolics and their inhibitory activity did not appear to be directly related to the resistance of a host or to the extent of its susceptibility to parasite infection.     

A survey on basal resistance and riboflavin-induced defense responses of sugar beet against Rhizoctonia solani

We examined basal defense responses and cytomolecular aspects of riboflavin-induced resistance (IR) in sugar beet-Rhizoctonia solani pathsystem by investigating H(2)O(2) burst, phenolics accumulation and analyzing the expression of phenylalanine ammonia-lyase (PAL) and peroxidase (cprx1) genes. Riboflavin was capable of priming plant defense responses via timely induction of H(2)O(2) production and phenolics accumulation. A correlation was found between induction of resistance by riboflavin and upregulation of PAL and cprx1 which are involved in phenylpropanoid signaling and phenolics metabolism. Application of peroxidase and PAL inhibitors suppressed not only basal resistance, but also riboflavin-IR of sugar beet to the pathogen. Treatment of the leaves with each inhibitor alone or together with riboflavin reduced phenolics accumulation which was correlated with higher level of disease progress. Together, these results demonstrate the indispensability of rapid H(2)O(2) accumulation, phenylpropanoid pathway and phenolics metabolism in basal defense and riboflavin-IR of sugar beet against R. solani.