887 resultados para Microwave induced resistance oscillations


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  果实为开花植物所特有的发育器官,在种子的成熟和传播过程中发挥着重要作用。同时,肉质果实中含有丰富的营养物质,包括纤维素、维生素、抗氧化剂等,成为人们饮食的重要组成部分。由于果实的成熟衰老和抗病性直接影响果品的质量和市场价值,因此,研究果实成熟衰老和抗病性的调控机制具有重要的理论意义和应用前景。本文主要利用蛋白质组学的方法,探讨外源化学物质抑制果实成熟衰老和诱导抗病性的调控机制。 1. 硅对果实的抗病性诱导:用硅酸钠(1%)处理采后的甜樱桃果实,再接种褐腐病原菌(Molinilia fracticola),置于20C下,观测贮藏期间果实的发病率,并分析硅处理后诱导的主要蛋白质及调控机制。研究结果表明:硅酸钠处理可显著抑制贮藏期间褐腐病的发生,其抑病机理与硅诱导PR-蛋白的表达,提高果实的抗氧化水平,减轻由病原菌侵染造成的氧化胁迫相关。同时,硅处理还能保护细胞骨架结构,有利于增强果实对病原菌入侵的抵抗力。 2. 水杨酸对果实的抗病性诱导:用水杨酸(SA,2mM)在果园处理三种成熟度的甜樱桃果实,然后接种青霉病原菌(Penicillium expansum)观察其发病情况,并取样分析参与抗病性应答的主要蛋白质及调控机制。试验结果表明:SA处理能显著降低青霉病的发病率和抑制病斑扩展,而且SA对低成熟度甜樱桃果实的抗性诱导效果更好。在八成熟的果实中,有5个热激蛋白和4个脱氢酶蛋白被SA诱导,这些蛋白参与了糖酵解和三羧酸循环。抗氧化蛋白和PR蛋白主要参与较低成熟度果实的抗性应答,而热激蛋白和脱氢酶在较高成熟度果实的抗性应答中更明显,SA诱导的抗性与代谢途径相关。   3. 草酸对果实的抗性诱导:用5mM的草酸处理冬枣果实后,接种青霉菌(P. expansum),观察果实发病情况,测定果实相关的生理指标,分析参与果实抗性应答的主要蛋白质及调控机制。结果表明:草酸能明显延缓冬枣果实的衰老,提高果实对青霉菌的抗性。草酸处理能抑制果实乙烯的释放量和呼吸强度,延缓叶绿素的降解,减少乙醇积累。利用蛋白质组学的研究方法证实了在25个参与了草酸处理应答的蛋白中,胱硫醚-β-合酶结构域包含蛋白(CBB domain-containing protein)和3个与光合作用相关蛋白[二磷酸核酮糖羧化酶/加氧酶(Ribulose bisphosphate carboxylase/oxygenase activase, chloroplast precursor),二磷酸核酮糖羧化酶/加氧酶大亚基结合蛋白(RuBisCO large subunit-binding protein subunit beta, chloroplast precursor),植物光系统Ⅱ放氧复合蛋白2(PSII oxygen-evolving complex protein 2)]的表达量上调,乙醇脱氢酶的表达量出现下调。草酸处理还提高了与乙烯合成前体相关蛋白的表达,抑制了ACC合成酶的活性。草酸提高果实抗病的机制与延缓果实成熟衰老和保持果实抗性有关。 4. 果实衰老的调控机制:采用高氧(100%)和低氧(2-3%)处理苹果果实,观察果实衰老的进程,并基于蛋白质组学的研究方法,探讨苹果果实衰老与线粒体蛋白质组的关系。结果表明,在苹果衰老过程中有22个蛋白的表达量发生变化,这些蛋白主要参与了三羧酸循环,电子传递,碳代谢和胁迫应答。高氧处理能诱导氧化胁迫,加速了果实的衰老。质谱鉴定结果证明:在高氧胁迫下,超氧化物歧化酶(manganese superoxide dismutase,MnSOD)和线粒体外膜通道蛋白(porin) 的表达量降低,MnSOD的活性受到抑制,由此提高了线粒体中超氧阴离子的含量,增加了蛋白质的氧化损伤。 此外,高氧处理改变了porin的功能,导致了线粒体膜的透势发生变化,从而引起外膜损伤。由此阐明了活性氧在果实的成熟衰老调控中的重要作用。

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在果实采后贮藏过程中,病原真菌的侵染会引起果实腐烂,造成巨大的经济损失。利用生物和非生物因子诱导果实抗病性,已经成为采后病害防治领域的一个研究热点。本文主要利用RT-PCR和RACE技术克隆果实抗病相关基因,通过分子杂交和蛋白羰基化免疫检测技术,研究了外源SA和酵母拮抗菌诱导果实抗病性机理,结果表明: 1. 通过优化RNA提取方法,能从含有多糖的冬枣、葡萄、甜樱桃、桃、番茄等果实中提取到质量较好的RNA,用于RT-PCR和Northern杂交。 2. 采用RT-PCR和RACE方法,从甜樱桃果实克隆了两个抗氧化相关基因CAT2(Genbank:EF165590)和GPX(Genbank:EF165591)和两个PR基因GLU-1(Genbank:EF177487)和GLU-3(Genbank:EF177488)。其中CAT2全长cDNA序列为1479 bp,编码492个氨基酸;GPX全长cDNA序列为513 bp,编码170个氨基酸;GLU-1全长cDNA序列为1050 bp,编码349个氨基酸;GLU-3部分cDNA序列为454 bp,编码141个氨基酸。 3. 酵母拮抗菌Pichia membranaefaciens处理不同成熟度的甜樱桃果实,能显著降低果实贮藏期间青霉病(Penicillium expansum)的发生,并且对低成熟度果实的病害防治效果更为明显。酵母拮抗菌的抑病机理与减轻了甜樱桃果实蛋白羰基化程度,诱导了果实抗氧化酶基因(CAT和GPX)和PR基因(GLU-1)的表达和提高了抗氧化酶(CAT和GPX)和β-1,3-葡聚糖酶的活性有关。 4. 四种酵母拮抗菌P. membranaefaciens, Cryptococcus laurentii, Candida guilliermondii和Rhodotorula glutinis处理桃果实,可显著降低贮藏期间的褐腐病(Monilinia fructicola)。这是由于酵母拮抗菌能抑制病原菌侵染造成的氧化胁迫和蛋白羰基化。此外,酵母拮抗菌处理还能显著诱导CAT、POD、几丁质酶、β-1,3-葡聚糖酶活性及相应基因的表达。 5. 水杨酸(SA,2 mM)处理采后不同成熟度的甜樱桃果实,能显著降低青霉病的危害。其抑病机理与SA处理能减轻P. expansum侵染引起的果实蛋白羰基化程度,显著提高CAT、GPX和β-1,3-葡聚糖酶基因的表达和相关的酶的活性有关。而2 mM的SA处理对P. expansum的生长没有直接抑制作用。 6. 水杨酸(SA,2 mM)与P. membranaefaciens(1×108 CFU/ml)配合处理能显著降低低温贮藏期间桃果实的褐腐病,并能提高几丁质酶、β-1,3-葡聚糖酶和POD的活性和相关基因的表达。另外,2 mM的SA对拮抗菌P. membranaefaciens的生长没有影响,但能够抑制病原菌M. fructicola的孢子萌发和菌丝扩展。

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目的: 利用重离子辐照技术选育出对农作物具有更好防病促生作用的突变生防菌株,探讨利用突变株诱导黄瓜对枯萎病菌产生抗病性的作用机制。 材料与方法: 采用兰州重离子研究装置(HIRFL)加速的碳离子束辐照生防菌BJ1,测定抑菌能力、抑菌谱,确定对该菌株最适宜的离子辐照参数,选育突变菌株。对突变株进行室内盆栽和田间防病促生试验。对原菌株和突变株进行16SrDNA和生理生化反应鉴定,确定分类地位。以突变株为对象进行诱导黄瓜对枯萎病菌产生抗病性的实验。 结果与结论: 1.重离子辐照生防菌BJ1的存活曲线随剂量的增加,呈先降后升再降的马鞍型变化,但是由于离子的能量不同也存在差异,表现为在相同的剂量下,能量越低其能量沉积效应即传能线密度(LET)越大,致死率越高。诱变效果随LET的不同也不尽相同,高LET时的突变株不但有更广的抑菌谱而且抑菌活性较对照也有比较大的提高,在存活率较高的条件下,低剂量就可以得到较多的突变体,有利于筛选优良的正突变体; 2.对于生防菌BJ1最适宜的12C6+辐照参数应选择剂量在200-400Gy,LET为60keV/m范围可筛选获得抑菌活性较高的菌株; 3.通过12C辐照结合抑菌试验最终获得了突变菌株154,该突变株通过20代的移植能够稳定遗传; 4.利用突变株154对黄瓜枯萎病菌进行室内盆栽促生试验,结果表明突变株154能够使促进黄瓜幼苗的生长发育,同时提高了黄瓜植株的抗病性,在对黄瓜枯萎病的防治效果上经154处理的达到了70.34%,高于原菌株和农药防治的效果; 5.传统的生理生化特征结合16SrDNA同源性比对的方法,对菌株BJ1及其突变株154的鉴定结果表明二者均属枯草芽孢杆菌,亲缘关系近,但是突变株154生化测定不同于原菌株,表现为抗菌物质的产量较高; 6.BJ1、154都可在番茄、当归和黄芪的根部有效的定殖,适应根部的生长环境, 并且154的定殖能力稍强; 7.BJ1、154防治当归麻口病效果较好,防治效果最好的是154的20倍液浸苗,防效为82.6%;BJ1的20倍液浸苗与154的10倍液浸苗对麻口病防治效果差别不大,防效分别为78.3%、75.62%;其余处理防效低于62%。 8.BJ1和154处理黄瓜幼苗后,植物体内一系列与抗病性有关的保护酶的活性均有不同程度的提高,因而可认为这些酶活性的改变与生防菌诱导的黄瓜对枯萎病的抗性可能有一定的相关性

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In marine benthic communities, herbivores consume a considerable proportion of primary producer biomass and, thus, generate selection for the evolution of resistance traits. According to the theory of plant defenses, resistance traits are costly to produce and, consequently, inducible resistance traits are adaptive in conditions of variable herbivory, while in conditions of constant/strong herbivory constitutive resistance traits are selected for. The evolution of resistance plasticity may be constrained by the costs of resistance or lack of genetic variation in resistance. Furthermore, resource allocation to induced resistance may be affected by higher trophic levels preying on herbivores. I studied the resistance to herbivory of a foundation species, the brown alga Fucus vesiculosus. By using factorial field experiments, I explored the effects of herbivores and fish predators on growth and resistance of the alga in two seasons. I explored genetic variation in and allocation costs of resistance traits as well as their chemical basis and their effects on herbivore performance. Using a field experiment I tested if induced resistance spreads via water-borne cues from one individual to another in relevant ecological conditions. I found that in the northern Baltic Sea F. vesiculosus communities, strength of three trophic interactions strongly vary among seasons. The highly synchronized summer reproduction of herbivores promoted their escape from the top-down control of fish predators in autumn. This resulted into large grazing losses in algal stands. In spring, herbivore densities were low and regulated by fish, which, thus,enhanced algal growth. The resistance of algae to herbivory increased with an increase in constitutive phlorotannin content. Furthermore, individuals adopted induced resistance when grazed and when exposed to water-borne cues originating from grazing of conspecific algae both in the laboratory and in field conditions. Induced resistance was adopted to a lesser extent in the presence of fish predators. The results in this thesis indicate that inducible resistance in F. vesiculosus is an adaptation to varying herbivory in the northern Baltic Sea. The costs of resistance and strong seasonality of herbivory have likely contributed to the evolution of this defense strategy. My findings also show that fish predators have positive cascading effects on F. vesiculosus which arise via reduced herbivory but possibly also through reduced resource allocation to resistance. I further found evidence that the spread of resistance via water-borne cues also occurs in ecologically realistic conditions in natural marine sublittoral. Thus, water-borne induction may enable macroalgae to cope with the strong grazing pressure characteristic of marine benthic communities. The results presented here show that seasonality can have pronounced effects on the biotic interactions in marine benthic communities and thereafter influence the evolution of resistance traits in primary producers.

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Introducción: Las vacunas clásicamente han representado un método económico y eficaz para el control y prevención de múltiples enfermedades infecciosas. En los últimos años se han introducido nuevas vacunas contra neumococo a precios elevados, y los diferentes análisis económicos a nivel mundial de estas vacunas no muestran tendencias. El objetivo de este trabajo era resumir la evidencia existente a través de los diferentes estudios económicos evaluando las dos vacunas de segunda generación contra neumococo en la población a riesgo. Metodología: En este trabajo se realizo una revisión sistemática de la literatura en 8 bases de datos localizadas en diferentes partes del mundo y también que tuvieran literatura gris. Los artículos fueron inicialmente evaluados acorde a su titulo y resumen, posteriormente los elegidos se analizaron en su totalidad. Resultados: Se encontraron 404 artículos, de los cuales 20 fueron incluidos en el análisis final. Se encontró que la mayoría de los estudios se realizaron en áreas donde la enfermedad tiene una carga baja, como es Norte América y Europa, mientras que en los lugares del mundo donde la carga es mas alta, se realizaron pocos estudios. De igual manera se observo que la mayoría de los estudios mostraron por los menos ser costo efectivos respecto a la no vacunación, y en su totalidad las dos vacunas de segunda generación mostraron costo efectividad respecto a la vacunación con PCV-7. Los resultados de los estudios son muy heterogéneos, hasta dentro del mismo país, señalando la necesidad de guías para la conducción de este tipo de estudios. De igual manera, la mayoría de los estudios fueron financiados por farmacéuticas, mientras en un numero muy reducido por entes gubernamentales. Conclusiones: La mayoría de los estudios económicos sobre las vacunas de segunda generación contra neumococo han sido realizados en países con un alto índice de desarrollo económico y patrocinados por farmacéuticas. Dado que la mayoría de la carga de la enfermedad se encuentran en regiones con un menor nivel de desarrollo económico se deberían realizar mas en estas zonas. De igual manera, al ser la vacunación un asunto de salud publica y con un importante impacto económico los gobiernos deberían estar mas involucrados en los mismos.

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Current research into indirect phytopathogen–herbivore interactions (i.e., interactions mediated by the host plant) is carried out in two largely independent directions: ecological/mechanistic and molecular. We investigate the origin of these approaches and their strengths and weaknesses. Ecological studies have determined the effect of herbivores and phytopathogens on their host plants and are often correlative: the need for long-term manipulative experiments is pressing. Molecular/cellular studies have concentrated on the role of signaling pathways for systemic induced resistance, mainly involving salicylic acid and jasmonic acid, and more recently the cross-talk between these pathways. This cross-talk demonstrates how interactions between signaling mechanisms and phytohormones could mediate plant–herbivore–pathogen interactions. A bridge between these approaches may be provided by field studies using chemical induction of defense, or investigating whole-organism mechanisms of interactions among the three species. To determine the role of phytohormones in induced resistance in the field, researchers must combine ecological and molecular methods. We discuss how these methods can be integrated and present the concept of “kaleidoscopic defense.” Our recent molecular-level investigations of interactions between the herbivore Gastrophysa viridula and the rust fungus Uromyces rumicis on Rumex obtusifolius, which were well studied at the mechanistic and ecological levels, illustrate the difficulty in combining these different approaches. We suggest that the choice of the right study system (possibly wild relatives of model species) is important, and that molecular studies must consider the environmental conditions under which experiments are performed. The generalization of molecular predictions to ecologically realistic settings will be facilitated by “middle-ground studies” concentrating on the outcomes of the interactions.

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Phytophagous insects have to contend with a wide variation in food quality brought about by a variety of factors intrinsic and extrinsic to the plant. One of the most important factors is infection by plant pathogenic fungi. Necrotrophic and biotrophic plant pathogenic fungi may have contrasting effects on insect herbivores due to their different infection mechanisms and induction of different resistance pathways, although this has been little studied and there has been no study of their combined effect. We studied the effect of the biotrophic rust fungus Uromyces viciae-fabae (Pers.) Schroet (Basidiomycota: Uredinales: Pucciniaceae) and the necrotrophic fungus Botrytis cinerea Pers. (Ascomycota: Helotiales: Sclerotiniaceae) singly and together on the performance of the aphid Aphis fabae Scop. (Hemiptera: Aphididae) on Vicia faba (L.) (Fabaceae). Alone, botrytis had an inhibitory effect on individual A. fabae development, survival and fecundity, while rust infection consistently enhanced individual aphids’ performance. These effects varied in linear relation to lesion or pustule density. However, whole-plant infection by either pathogen resulted in a smaller aphid population of smaller aphids than on uninfected plants, indicating a lowering of aphid carrying capacity with infection. When both fungi were applied simultaneously to a leaf they generally cancelled the effect of each other out, resulting in most performance parameters being similar to the controls, although fecundity was reduced. However, sequential plant infection (pathogens applied five days apart) led to a 70% decrease in fecundity and 50% reduction in intrinsic rate of increase. The application of rust before botrytis had a greater inhibitory effect on aphids than applying botrytis before rust. Rust infection increased leaf total nitrogen concentration by 30% while infection by botrytis with or without rust led to a 38% decrease. The aphids’ responses to the two plant pathogens individually is consistent with the alteration in plant nutrient content by infection and also the induction of different plant defence pathways and the possible cross-talk between them. This is the first demonstration of the complex effects of the dual infection of a plant by contrasting pathogens on insect herbivores. Key words: Vicia faba, Botrytis cinerea, Uromyces viciae-fabae, tripartite interactions, induced resistance

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Objetivando-se estudar a influência da adubação na manifestação da resistência de feijoeiro (Phaseolus vulgaris L.) (cultivares Rosinha G-2, ESAL-564 - Carioca e Diamante Negro) ao ataque de Acanthoscelides obtectus (Say, 1831) (Coleoptera: Bruchidae) foram conduzidos, na época das águas, testes com e sem chance de escolha, utilizando-se grãos obtidos de parcelas adubadas com N, P, K, NP, NK, PK, NPK, e sem adubo, totalizando 24 tratamentos. Os testes foram realizados em condições controladas de temperatura, umidade e luz. Foram avaliados o número de insetos atraídos e de ovos, em cada tratamento. Concluiu-se, no teste com chance de escolha, que o número de ovos de A. obtectus por recipiente foi reduzido pela aplicação do nitrogênio. A aplicação de nitrogênio em adubação resultou em menor porcentagem de insetos atraídos e menor número de ovos no genótipo Rosinha G-2. A manifestação da resistência nos genótipos ESAL-564 e Rosinha G-2 ao ataque de A. obtectus ficou evidente quando utilizados nitrogênio e potássio. Nos testes sem chance de escolha o consumo dos insetos foi reduzido nos grãos produzidos com a aplicação do nitrogênio. A aplicação de nitrogênio em adubação resultou no aumento do número de ovos de A. obtectus no genótipo ESAL-564. Não ficou evidente a manifestação da resistência nos genótipos ESAL-564 e Rosinha G-2 ao ataque do caruncho, pela aplicação dos macronutrientes N, P e K. Na ausência de fósforo o ciclo biológico do inseto foi maior na presença de potássio.

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Perdas significativas ocorrem durante o armazenamento e a comercialização de uvas de mesa devido, principalmente, à ocorrência do mofo cinzento (Botrytis cinerea Pers.:Fr.) e, para o controle de patógenos emprega-se, geralmente, o dióxido de enxofre (SO2). Diante da restrição crescente ao uso de produtos químicos em pós-colheita, tem ocorrido considerável interesse em métodos alternativos de controle. Este trabalho teve como principal objetivo avaliar os efeitos da quitosana, na proteção pós-colheita de uva 'Itália' contra B. cinerea. In vivo, avaliou-se o efeito direto e indireto da quitosana pelo tratamento dos cachos de uva, antes e após a inoculação com o patógeno. Utilizou-se quitosana nas concentrações de 0,00; 0,25; 0,50; 1,00; 1,50 e 2,00 % (v/v). Para inoculação, em 10 bagas de cada cacho de uva foram feitos ferimentos de ±2 mm de profundidade, procedendo-se em seguida, a aspersão da suspensão de conídios (±10(5) conídios.mL-1) de B. cinerea. Após os tratamentos, os cachos foram mantidos a 25±1 °C / 80-90 % UR e avaliados diariamente quanto à incidência e severidade da podridão. Avaliações in vitro do efeito do produto sobre o patógeno também foram realizadas analisando-se o crescimento micelial e a germinação dos conídios de B. cinerea. A solução de quitosana, nas concentrações de 1,5 e 2,0 % (v/v), quando empregada após a inoculação com B cinerea, reduziu significativamente o índice de doença no entanto, quando os cachos foram tratados antes da inoculação, não houve efeito significativo do tratamento sobre o desenvolvimento da doença. Nos ensaios in vitro, a solução de quitosana, nas maiores concentrações, suprimiu o crescimento micelial do patógeno e retardou a germinação dos conídios.

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The role of non-pathogenic binucleate Rhizoctonia spp. (BNR) on the biocontrol of diseases caused by R. solani on many crops has been reported in the literature. However, in Brazil, there is no information about the potential of BNR as biocontrol agents against Rhizoctonia diseases on soybean. on this research we tested the hypothesis that BNR can induce resistance on soybean against the foliar blight caused by R. solani anastomosis group (AG) 1 IA. Thus, the objective of this research was to evaluate BNR isolates isolated from peanuts, snapbeans and soybean according to their ability for inducing resistance on soybean against the foliar blight disease, under greenhouse conditions. This research evidenced the role of BNR inducing resistance on soybeans against the foliar blight. However, both the occurrence and effectiveness of the phenomenon of induced resistance are dependent on the soybean cultivation season.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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O tripes do prateamento, Enneothrips flavens Moulton, 1941, é considerado a principal praga do amendoim no Brasil, por sua ocorrência generalizada, pelos elevados níveis populacionais e pelos danos causados a cultura. Objetivou-se, com este trabalho, avaliar o efeito do silício sobre a população de E. flavens e sobre a produtividade do amendoinzeiro. Avaliaram-se, semanalmente, 10 folíolos abertos ou semi-abertos no terço superior das plantas do cultivar IAC 886, por parcela. Os tratamentos foram constituídos por: uma aplicação foliar de silício realizada aos 20 dias após a emergência, duas aplicações foliares de silício realizadas aos 20 e 55 dias, e o controle. Uma aplicação de silício proporcionou proteção às plantas de amendoim, reduzindo o número de adultos e ninfas do tripes de E. flavens e aumentou a produtividade da cultura em 31,30% de amendoim em casca e 28,85% em grãos.

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Barley plants (cultivars Embrapa 127, 128 and 129) treated with xanthan gum, and with different time intervals between the administration of the inducer and the pathogen, demonstrated induction of resistance against Bipolaris sorokiniana. Induction was shown to have local and systemic action. In order to prove the resistance effect, biochemical analyses were performed to quantify proteins and the enzymatic activity of beta-1,3 glucanase. Results demonstrated that barley plants treated with the inducer, showed an increase in the concentration of proteins, as well as in the activity of the enzyme beta-1,3 glucanase, when compared with the extract from healthy plants. In infected plants, protein concentrations decreased and enzymatic activity was lower than in healthy plants. Results suggest that barley plants treated with xanthan gum developed mechanisms responsible for induced resistance, which are still unknown. The most important macromolecule in the defense mechanism was demonstrated to be PR-protein, due to its accumulation and concentration of proteins. However, it may not be the only macromolecule responsible for the resistance effect. (C) 2004 Elsevier SAS. All rights reserved.