During avocado fruit ripening, decreasing levels of the flavonoid epicatechin have been reported to modulate the metabolism of preformed antifungal compounds and the activation of quiescent Colletotrichum gloeosporioides infections. Epicatechin levels decreased as well when C. gloeosporioides was grown in the presence of epicatechin in culture. Extracts of laccase enzyme obtained from decayed tissue and culture media fully metabolized the epicatechin substrate within 4 and 20 h, respectively. Purified laccase protein from C. gloeosporioides showed an apparent MW of 60,000, an isoelectric point at pH 3.9, and maximal epicatechin degradation at pH 5.6. Inhibitors of fungal laccase such as EDTA and thioglycolic acid reduced C. gloeosporioides symptom development when applied to ripening susceptible fruits. Isolates of C. gloeosporioides with reduced laccase activity and no capability to metabolize epicatechin showed reduced pathogenicity on ripening fruits. On the contrary, Mexican isolates with increasing capabilities to metabolize epicatechin showed early symptoms of disease in unripe fruits. Transcript levels of cglac1, encoding C. gloeosporioides laccase, were enhanced during fungal development in the presence of epicatechin at pH 6.0, where avocado fruits are susceptible to fungal attack. But transcript increase was not detected at pH 5.0, where the fruit is resistant to fungal attack. The present results suggest that biotransformation of epicatechin by C. gloeosporioides in ripening fruits is followed by the decline of the preformed antifungal diene compound, resulting in the activation of quiescent infections.
In recent years, anthracnose has become a significant disease affecting avocado fruit in the state of Michoacan, Mexico, where it significantly reduces fruit quality and commercial yield. Anthracnose has been assumed to involve Colletotrichum gloeosporioides and C. acutatum as causal agents. However, because of the increasing incidence of anthracnose, a more precise identification of the Colletotrichum spp. involved in this disease has become desirable. During the years 2004-2007, avocado fruits of different sizes exhibiting brown-black and reddish spots on the pericarp and soft rot in the mesocarp, were gathered from orchards in nine counties. Fungal isolates were cultured on potato dextrose agar, and among these, 31 were selected for molecular, morphological and pathogenicity analyses. The molecular approaches used sequence typing of the internal transcribed spacer region and the partial nuclear large ribosomal subunit, allowing the unequivocal identification of C. gloeosporioides (71%), C. acutatum (16%) and C. boninense (13%). This last species has not been previously reported as being associated with anthracnose symptoms in avocado fruits anywhere in the world. Various morphological characteristics such as the size and shape of conidia were determined, as well as the conidial mass colour. Pathogenicity tests performed with all three species were conducted by inoculating healthy fruits. In each case, identical symptoms developed within 3 days of inoculation. Knowledge of the Colletotrichum populations in the Michoacan state, including the newly encountered avocado pathogen C. boninense, will facilitate further studies addressing the relationships between these Colletotrichum spp. and their avocado host.
This review presents information on disease outbreaks in human populations linked to Salmonella spp. and Escherichia coli O157:H7 associated with the consumption of fresh produce. It focuses on the processes of bacterial internalization and survival in non-host plant tissues and the role of biofilms in bacterial persistence. Research to identify the microbial sources contaminating fruits and vegetables and their persistence is urgently required. Internalization of human pathogenic bacteria in plant tissues is reported for various fruits. Under certain circumstances these can enter the plant via roots or seeds, with subsequent survival and translocation within the plant. Research should continue on the relationships between human pathogens and plants, and treatments should be developed to minimize pathogen presence on the surface of raw produce and to prevent their internalization.
During the spring of 2015, fruit rot symptoms were observed on pomegranate fruit in an orchard located in Oaxaca, Mexico. Characteristic lesions were collected and a fungus was isolated from these lesions. Based on morphological characteristics, analysis of rDNA-ITS sequences, and pathogenicity tests on pomegranate fruits, the causal agent was identified as Pilidiella granati. This is the first report of P. granati causing pomegranate fruit rot in Mexico.
Zea mays and Sorghum bicolor are important crops for animal and human nutrition worldwide. In the Central Highland Valley of Mexico, both crops are extremely important, and research is aimed toward increasing yield, disease resistance, and crop adaptation from 1,900- to 2,700-m elevation. In a 3-year field breeding experiment (2004 to 2006), leaf blight and vascular wilt symptoms were frequently observed in contiguous plots of maize and sorghum crops in Montecillo, Mexico and maize plots in Tecamac, Mexico. To identify and characterize the causal agent of these symptoms, isolations were conducted on leaves from areas where healthy and diseased tissues converged. Leaf sections of 1 cm2 from both crops were disinfested, placed on casamino acid-peptone-glucose (CPG) medium, and incubated at 28°C. After 48 h, only yellow colonies were observed and 12 isolates were selected for further characterization. Physiological and biochemical tests indicated that the isolates were nonfluorescent on King's B medium, and API 50 CHE (bioMérieux, Marcy l'Etoile, France) revealed that they were negative for gelatin hydrolysis, indole production, acid production from raffinose and positive for utilization of glycerol, D-glucose, mannitol, arbutine, esculine, salicine, cellobiose, maltose, melibiose, D-fucose, and D-arabitol; all characteristics of Pantoea agglomerans. Further identification of these isolates was accomplished by DNA analysis. For DNA analysis, 1.4-kbp fragments of the 16S rRNA gene were amplified with primer set 8F/1492R (3) and sequenced with U514F/800R universal primers (2). Five sequences were obtained and deposited in GenBank (Accession Nos. EF050806 to EF050810). A phylogenetic tree was constructed using the UPGMA method (mega version 3.1). Results of the phylogenetic analysis grouped the species P. ananatis, P. stewartti, and P. agglomerans into three clusters. The five unknown sequences were grouped into the P. agglomerans cluster. There was a 98 to 99% similarity of the five 16S rRNA gene sequences with P. agglomerans strain type ATCC 27155. Pathogenicity of the 12 isolates was confirmed by injecting 108 CFU mL–1 of inoculum into stems of 3-week-old maize cv. Triunfo and sorghum cold tolerant hybrid (A1×B5)×R1 seedlings in the greenhouse at 28°C and 80% relative humidity. Also, seedlings were inoculated with water, nonpathogenic isolates of P. agglomerans from maize (GM13, and HLA1), and not inoculated as negative controls. Three replications were included for each isolate and control. All test strains developed water-soaked lesions on juvenile leaves at 8 days postinoculation and were followed by chlorotic to straw-colored leaf streaks and then leaf blight symptoms at 3 weeks postinoculation. All negative control seedlings did not develop symptoms. In addition, the 12 isolates were infiltrated at 107 CFU mL–1 into tobacco leaves that displayed a hypersensitive response at 4 days, indicating the presence of the type III secretion system (1). Isolates were reisolated, and the 16S rRNA gene fragments were 100% similar to their original isolate sequences. P. agglomerans has been reported to affect other crops, including chinese taro in Brazil (2007), onion in the United States (2006) and South Africa (1981), and pearl millet in Zimbabwe (1997); however, to our knowledge, this is the first report of P. agglomerans associated with leaf blight and vascular wilt symptoms in maize and sorghum in the Central Highland Valley of Mexico. References: (1) J. Alfano and A. Collmer. Annu. Rev. Phytopathol 42:385, 2004. (2) Y. Anzai et al. Int. J. Syst. Evol. Microbiol. 50:1563, 2000. (3) M. Sasoh et al. Appl. Environ. Microbiol. 72:1825, 2006.
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