Development of biological control for plant diseases is accepted as a durable and environmentally friendly alternative for agrochemicals. Arbuscular mycorrhizal fungi (AMF), which form symbiotic associations with root systems of most agricultural, horticultural and hardwood crop species, have been suggested as widespread potential bioprotective agents. In the present study the ability of two AMF (Glomus mosseae and Glomus intraradices) to induce local or systemic resistance to Phytophthora parasitica in tomato roots have been compared using a split root experimental system. Glomus mosseae was effective in reducing disease symptoms produced by P. parasitica infection, and evidence points to a combination of local and systemic mechanisms being responsible for this bioprotector effect. The biochemical analysis of different plant defence-related enzymes showed a local induction of mycorrhiza-related new isoforms of the hydrolytic enzymes chitinase, chitosanase and beta-1,3-glucanase, as well as superoxide dismutase, an enzyme which is involved in cell protection against oxidative stress. Systemic alterations of the activity of some of the constitutive isoforms were also observed in non-mycorrhizal roots of mycorrhizal plants. Studies on the lytic activity against Phytophthora cell wall of root protein extracts also corroborated a systemic effect of mycorrhizal symbiosis on tomato resistance to Phytophthora.
The arbuscular mycorrhizal fungus Glomus mosseae is able to confer bioprotection against Phytophthora parasitica in tomato roots. Localized and induced systemic resistance (ISR) have been demonstrated to be involved in pathogen control in mycorrhizal and nonmycorrhizal roots with a split root experimental system. Decreased pathogen development in mycorrhizal and nonmycorrhizal parts of mycorrhizal root systems is associated with accumulation of phenolics and plant cell defense responses. G. mosseae-containing cortical cells in the mycorrhizal tissues are immune to the pathogen and exhibit a localized resistance response with the formation of cell wall appositions reinforced by callose adjacent to intercellular hyphae. The systemically induced resistance in nonmycorrhizal root parts is characterized by elicitation of host wall thickenings containing non-esterified pectins and PR-1a protein in reaction to intercellular pathogen hyphae, and by the formation of callose-rich encasement material around P. parasitica hyphae that are penetrating root cells. PR-la protein is detected in the pathogen wall only in these tissues. None of these cell reactions are observed in nonmycorrhizal pathogen-infected root systems, where disease development leads to host cell death. The cellular and molecular basis of bioprotection by an arbuscular mycorrhizal fungus is discussed in relation to that induced by other nonpathogenic microorganisms.
Tomato plants pre-colonised by the arbuscular mycorrhizal fungus Glomus mosseae showed decreased root damage by the pathogen Phytophthora nicotianae var. parasitica. In analyses of the cellular bases of their bioproteetive effect, a prerequisite for cytological investigations of tissue interactions between G. mosseae and P. nicotianae v. parasitica was to discriminate between the hyphae of the two fungi within root tissues. We report the use of antibodies as useful tools, in the absence of an appropriate stain for distinguishing hyphae of P. nicotianae v. parasitica from those of G. mosseae inside roots, and present observations on the colonisation patterns by the pathogenic fungus alone or during interactions in mycorrhizal roots. Infection intensity of the pathogen, estimated using an immunoenzyme labelling technique on whole root fragments, was lower in mycorrhizal roots. Immunogold labelling of P. nicotianae v. parasitica on cross-sections of infected tomato roots showed that inter or intracellular hyphae developed mainly in the cortex, and their presence induced necrosis of host cells, the wall and contents of which showed a strong autofluorescence in reaction to the pathogen. In dual fungal infections of tomato root systems, hyphae of the symbiont and the pathogen were in most cases in different root regions, but they could also be observed in the same root tissues. The number of P. nicotianae v. parasitica hyphae growing in the root cortex was greatly reduced in mycorrhizal root systems, and in mycorrhizal tissues infected by the pathogen, arbuscule-containing cells surrounded by intercellular P. nicotianae v. parasitica hyphae did not necrose and only a weak autofluorescence was associated with the host cells. Results are discussed in relation to possible processes involved in the phenomenon of bioprotection in arbuscular mycorrhizal plants.
Fusarium oxysporum includes nonpathogenic strains and pathogenic strains that can induce necrosis or tracheomycosis in plants. The objective of this study was to compare the abilities of a pathogenic strain (Foln3) and a nonpathogenic strain (Fo47) to colonize flax roots and to induce early physiological responses in flax cell culture suspensions. Both strains colonized the outer cortex of the root; however, plant defense reactions, i.e., the presence of wall appositions, osmiophilic material, and collapsed cells, were less frequent and less intense in a root colonized by Foln3 than by Fo47. Early physiological responses were measured in flax cell suspensions confronted with germinated microconidia of both strains. Both pathogenic (Foln3) and nonpathogenic strains (Fo47) triggered transient H 2 O 2 production in the first few minutes of the interaction, but the nonpathogenic strain also induced a second burst 3 h postinoculation. Ca 2؉ influx was more intense in cells inoculated with Fo47 than in cells inoculated with Foln3. Similarly, alkalinization of the extracellular medium was higher with Fo47 than with Foln3.
Transgenic mice expressing the prion protein (PrP) of species affected by transmissible spongiform encephalopathies (TSEs) have recently been produced to facilitate experimental transmission of these diseases by comparison with wild-type mice. However, whilst wild-type mice have largely been described for the discrimination of different TSE strains, including differentiation of agents involved in bovine spongiform encephalopathy (BSE) and scrapie, this has been only poorly described in transgenic mice. Here, two ovine transgenic mouse lines (TgOvPrP4 and TgOvPrP59), expressing the ovine PrP (A136 R154 Q171) under control of the neuron-specific enolase promoter, were studied; they were challenged with brainstem or spinal cord from experimentally BSE-infected sheep (AA136 RR154 QQ171 and AA136 RR154 RR171 genotypes) or brainstem from cattle BSE and natural sheep scrapie. The disease was transmitted successfully from all of these sources, with a mean of approximately 300 days survival following challenge with material from two ARQ-homozygous BSE-infected sheep in TgOvPrP4 mice, whereas the survival period in mice challenged with material from the ARR-homozygous BSEinfected sheep was 423 days on average. It was shown that, in the two ovine transgenic mouse lines, the Western blot characteristics of protease-resistant PrP (PrP res ) were similar, whatever the BSE source, with a low apparent molecular mass of the unglycosylated glycoform, a poor labelling by P4 monoclonal antibody and high proportions of the diglycosylated form. With all BSE sources, but not with scrapie, florid plaques were observed in the brains of mice from both transgenic lines. These data reinforce the potential of this recently developed experimental model for the discrimination of BSE from scrapie agents.
INTRODUCTIONTransmissible spongiform encephalopathies (TSEs) are fatal, neurodegenerative diseases of the central nervous system (CNS) affecting both humans (Creutzfeldt-Jakob disease; CJD) and animals, mainly sheep and goats (scrapie), deer and elk (chronic wasting disease), and cattle (bovine spongiform encephalopathy; BSE). Incubation periods are long and ultimately lead to fatal neurological clinical signs. The disease is generally associated with the accumulation of an abnormal form of a host-encoded prion protein (PrP c ) in the CNS (Prusiner, 1982). This disease-associated PrP (PrPd) differs from the cellular protein (PrP c ) in its biochemical properties, including partial resistance to proteinase K degradation (PrP res ) and insolubility in nondenaturing detergents. Prion-disease transmission between different species, if it occurs, is generally characterized by prolonged incubation periods and low susceptibility at first passage in a new host species. On subsequent subpassages in the new host species, the infectious agent generally adapts to give shorter incubation times and higher susceptibility. This illustrates the existence of a 'species barrier' phenomenon. The transmission efficiency of the disease across species largely depend...
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