Genetic diversity among viruses associated with sugarcane mosaic disease in Tucumán, Argentina.

Sugarcane leaves with mosaic symptoms were collected in 2006--07 in Tucumán (Argentina) and analyzed by reverse-transcriptase polymerase chain reaction (RT-PCR) restriction fragment length polymorphism (RFLP) and sequencing of a fragment of the Sugarcane mosaic virus (SCMV) and Sorghum mosaic virus (SrMV) coat protein (CP) genes. SCMV was detected in 96.6% of samples, with 41% showing the RFLP profile consistent with strain E. The remaining samples produced eight different profiles that did not match other known strains. SCMV distribution seemed to be more related to sugarcane genotype than to geographical origin, and sequence analyses of CP genes showed a greater genetic diversity compared with other studies. SrMV was detected in 63.2% of samples and most of these were also infected by SCMV, indicating that, unlike other countries and other Argentinean provinces, where high levels of co-infection are infrequent, co-existence is common in Tucumán. RFLP analysis showed the presence of SrMV strains M (68%) and I (14%), while co-infection between M and H strains was present in 18% of samples. Other SCMV subgroup members and the Sugarcane streak mosaic virus (SCSMV) were not detected. Our results also showed that sequencing is currently the only reliable method to assess SCMV and SrMV genetic diversity, because RT-PCR-RFLP may not be sufficiently discriminating.

Effect of fragarin on the cytoplasmic membrane of the phytopathogen Clavibacter michiganensis.

Fragarin, an antibiotic that was isolated and purified from a soluble fraction of strawberry leaves, may be a new type of preformed antimicrobial compound (phytoanticipin). Here, we report that the growth and oxygen consumption of the phytopathogenic bacterium Clavibacter michiganensis were rapidly inhibited after the addition of fragarin to cultures. Also, dissipation of the membrane potential and an increase of cell membrane permeability were observed in the presence of fragarin. The ability of fragarin to dissipate the membrane potential was confirmed with the use of small unilamellar liposomes made with lipids extracted from C. michiganensis. Our results suggest that fragarin is able to act at the membrane level, and that this action is correlated with a decrease in cell viability.

First Report of Colletotrichum acutatum on Strawberry in Northwestern Argentina.

Isolates were obtained from strawberry tissue with anthracnose symptoms from several locations near Tucumán, Argentina. Isolates were characterized using several criteria. Isolates produced fusiform conidia, tapered to a point at both ends, and averaged 13.5 × 4.9 µm. On potato dextrose agar, colonies produced a white cottony mycelial colony that turned orange in older cultures. Compared with Colletotrichum fragariae, the new isolates produced fewer appressoria. Pathogenicity tests were conducted on detached leaves and plants in the greenhouse and field. Detached immature leaves of cvs. Chandler, Fern, and Sweet Charlie were inoculated with a 20-µl droplet of an aqueous conidial suspension (106 conidia per ml) placed on the adaxial surface. Control leaves were inoculated with sterile distilled water. Leaves were maintained under white light (2,000 lux, 12 h/day) at 26°C, and 100% relative humidity. Necrotic spots were visible 4 days after inoculation. Greenhouse and field plants were spray-inoculated and covered for 48 h. Disease symptoms were mainly observed on petioles and runners 9 days after inoculation. No lesions were observed on control detached leaves or plants. Koch's postulates were confirmed in all cases. Based on morphological and cultural characteristics, isolates were identified as C. acutatum Simmonds (1). This is the first report of C. acutatum causing strawberry anthracnose in northwestern Argentina. Reference: (1) B. Smith and L. L. Black. Plant Dis. 74:69, 1990.

Isolation and purification of a 316 Da preformed compound from strawberry (Fragaria ananassa) leaves active against plant pathogens.

An antibiotic called fragarin showing activities against bacterial and fungal plant pathogens was isolated and purified by FPLC chromatography from the soluble fraction of strawberry leaves. The molecular weight value determined by mass spectrometry is 316 Da. Fragarin remains fully active after protease treatment or alkaline hydrolysis at 100 degrees C for 20 min. Biological and chemical analyses suggest that fragarin may be a new type of an antimicrobial preformed compound--phytoanticipin--and would constitute a primary non-specific barrier of strawberry defense.