<Emphasis Type="Italic">Stenotrophomonas maltophilia</Emphasis>: a new potential biocontrol agent of <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis>, causal agent of potato brown rot

Stenotrophomonas maltophilia was isolated from the rhizosphere of eggplant in the Nile Delta of Egypt, and its antagonistic potential against Ralstonia solanacearum race 3 biovar 2, the causal agent of potato brown rot, was in vitro evaluated on KB agar medium and in vivo on potato plants. In vitro, four isolates of S. maltophilia (PD3531, PD3532, PD3533, and PD3534) appeared antagonistic. The isolate (PD3533) was screened as the most promising antagonist for the in vivo tests. In the greenhouse, the antagonist was applied directly to soil or by bacterization of potato eyepieces. Stenotrophomonas maltophilia significantly suppressed potato brown rot in Egyptian clay soil but not in Dutch clay soil. Survival of a rifampicin and chloramphenicol-resistant S. maltophilia strain PD4560 was investigated in two pairs of clay soils, conventionally and organically managed, from Egypt and the Netherlands. The survival of S. maltophilia was significantly less in Dutch than in Egyptian soils, while the converse occurred for R. solanacearum. These results are in agreement with those obtained in the in vivo biocontrol tests. In conclusion, S. maltophilia may be useful for control of brown rot in the area where it was originally isolated, the Nile Delta in Egypt.     

Lack of biocontrol capacity in a non-pathogenic mutant of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">melonis</Emphasis>

The aim of this study was to assess the biocontrol capacity of rev157, a non-pathogenic mutant of a pathogenic strain of Fusarium oxysporum f. sp. melonis (Fom24). Inoculated in association with the virulent parental strain, the mutant rev157 did not protect the host plant (muskmelon) against infection by Fom24. Applied on flax, a non-host plant, the mutant rev157 was not able to protect it against its specific pathogen F. oxysporum f. sp. lini. On the contrary the parental strain Fom24 did protect flax as well as a soil-borne biocontrol strain (Fo47). Since the mutant rev157 was affected neither in its growth in vitro nor in its capacity to penetrate into the roots, it can be speculated that the mutation has affected traits responsible for interactions within the plant. In F. oxysporum the pair of strains Fom24/rev157 is a good candidate to identify genes involved in the biocontrol capacity of F. oxysporum and to test the hypothesis of a link between capacity to induce the disease and capacity to induce resistance in the plant.     

<Emphasis Type="Italic">In vitro</Emphasis> screening for sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis>L.) resistant calli to <Emphasis Type="Italic">Diaporthe helianthi</Emphasis> fungal culture filtrate

Diaporthe helianthi the causal agent of sunflower (Helianthus annuus) stem canker, causes significant reductions in yield and oil content in most sunflower-growing areas. With the aim of enhancing host resistance, we selected in vitro sunflower calli against culture filtrates of two pathogen isolates (7/96 and 101/96). This technique may be an effective and rapid tool to discriminate the most virulent D. helianthi isolate and to screen for host resistance in the early stage of a breeding programme. Further investigation on the mechanisms involved in defence pathways showed no induction of salicylic acid and pathogenesis-related proteins in calli, indicating that the host resistance is not associated with Systemic Acquired Resistance but probably other biochemical mechanisms.     

Root and stem rot of chrysanthemum caused by five <Emphasis Type="Italic">Pythium</Emphasis> species in Japan

Root and stem rot with wilt of above ground parts of cultivated chrysanthemums was first found in Ibaraki, Toyama and Kagawa prefectures, Japan in 2002 and 2003. Pythium species were isolated from the diseased tissues and identified as P. dissotocum, P. oedochilum, P. sylvaticum, P. ultimum var. ultimum and asexual strains of P. helicoides based on their morphologies and sequences of rDNA-ITS region. All the Pythium species were strongly pathogenic to chrysanthemums in pot conditions and were reisolated from the inoculated plants. Because Pythium root and stem rot of chrysanthemum has never been reported in Japan, we propose that this is a new disease that can be caused by the five Pythium species.     

Effect of indole-acetic acid (IAA) on the development of symptoms caused by <Emphasis Type="Italic">Pythium ultimum</Emphasis> on tomato plants

The effect of indole-acetic acid (IAA) on the development of symptoms caused by Pythium ultimum on tomato plants was investigated using different bioassays. Application of IAA (5 μg ml⁻¹) on tomato seedlings inoculated with P. ultimum did not affect their emergence suggesting that IAA did not affect the severity of Pythium damping-off. However, IAA was shown to influence the development of P. ultimum symptoms on tomato plantlets. Low concentrations of IAA (0–0.1 μg ml⁻¹) within the rhizosphere of plantlets increased the severity of the symptoms caused by P. ultimum, while higher concentrations (10 μg ml⁻¹), applied either by drenching to the growing medium or by spraying on the shoot, reduced the symptoms caused by this pathogen. In addition, the study demonstrated that P. ultimum produces IAA in liquid culture amended with L-tryptophan, tryptamine or tryptophol (200 μg ml⁻¹) and in unamended culture.     

Molecular characterization of <Emphasis Type="Italic">Phoma tracheiphila</Emphasis>, causal agent of Mal secco disease of citrus,in Israel

Mal secco disease of citrus caused by Phoma tracheiphila is a devastating disease in the Mediterranean basin. Susceptible citrus species include lemon, citron, lime and others. Trees attacked by the fungus show characteristic symptoms; the smallest twigs die first, followed by the larger branches. Eventually, the whole tree is killed. The symptoms are clear in the orchards but by the time they are visible the disease is already well established. The need for a sensitive, reliable and rapid diagnostic method for the early identification of the fungus in trees and fruit exists. We have developed a PCR-based method for the identification of P. tracheiphila from plant tissues including fruit. Any such method must take into account the genetic variability in the pathogen population. Molecular methods were used to compare different isolates of P. tracheiphila. This study found no significant differences between different isolates from different citrus species from different parts of Israel.     

Cloning of DNA fragments specific to the pathotype and race of <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Japanese isolates of Verticillium dahliae, a causal agent of wilt disease in many plants, are classifiable into pathotypes based on their pathogenicity. Because these pathotypes are morphologically indistinguishable, establishing a rapid identification method is very important for the control of this pathogen in Japan. For cloning DNA fragments that are useful for identification and specific detection of V. dahliae pathotypes, we performed random amplified polymorphic DNA (RAPD) analyses using various isolates. One polymerase chain reaction (PCR) product, E10-U48, was specific to isolates pathogenic to sweet pepper. The other product, B68-TV, was specific to race 1 of isolates pathogenic to tomato. The specificity of these sequences was confirmed by genomic Southern hybridization. Further analyses revealed that the region peripheral to B68-TV obtained from the genomic DNA library includes the sequence specific to all isolates pathogenic to tomato (races 1 and 2). Moreover, sequence tagged site (STS) primers designed from B68-TV and its peripheral region showed race-specific and pathotype-specific amplification in a PCR assay. The probes and primers obtained in this study are likely to be useful tools for the identification and specific detection of pathotypes and races of V. dahliae. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession number AB095266.