Transcriptional Regulation of Metabolic and Cellular Processes in Durum Wheat (Triticum turgidum subsp. durum) in the Face of Temperature Increasing.

The Yaqui Valley, Mexico, has been historically considered as an experimental field for semiarid regions worldwide since temperature is an important constraint affecting durum wheat cultivation. Here, we studied the transcriptional and morphometrical response of durum wheat at an increased temperature (+2 °C) for deciphering molecular mechanisms involved in the thermal adaptation by this crop. The morphometrical assay showed a significant decrease in almost all the evaluated traits (shoot/root length, biovolume index, and dry/shoot weight) except in the dry root weight and the root:shoot ratio. At the transcriptional level, 283 differentially expressed genes (DEGs) were obtained (False Discovery Rate (FDR) ≤ 0.05 and |log2 fold change| ≥ 1.3). From these, functional annotation with MapMan4 and a gene ontology (GO) enrichment analysis with GOSeq were carried out to obtain 27 GO terms significantly enriched (overrepresented FDR ≤ 0.05). Overrepresented and functionally annotated genes belonged to ontologies associated with photosynthetic acclimation, respiration, changes in carbon balance, lipid biosynthesis, the regulation of reactive oxygen species, and the acceleration of physiological progression. These findings are the first insight into the regulation of the mechanism influenced by a temperature increase in durum wheat.

Metabolic Footprints of Burkholderia Sensu Lato Rhizosphere Bacteria Active against Maize Fusarium Pathogens.

Consistent with their reported abundance in soils, several Burkholderia sensu lato strains were isolated from the rhizosphere of maize plants cultivated at different sites in central México. Comparative analysis of their 16S rRNA gene sequences permitted their separation into three distinctive clades, which were further subdivided into six other clusters by their close resemblance to (1) Trinickia dinghuensis; (2) Paraburkholderia kirstenboschensis, P. graminis, P. dilworthii and P. rhynchosiae; (3) B. gladioli; (4) B. arboris; (5) B. contaminans, or (6) B. metallica representative species. Direct confrontation assays revealed that these strains inhibited the growth of pathogenic Fusarium oxysporum f. sp. radicis-lycopersici, and F. verticillioides within a roughly 3-55% inhibition range. The use of a DIESI-based non-targeted mass spectroscopy experimental strategy further indicated that this method is an option for rapid determination of the pathogen inhibitory capacity of Burkholderia sensu lato strains based solely on the analysis of their exometabolome. Furthermore, it showed that the highest anti-fungal activity observed in B. contaminans and B. arboris was associated with a distinctive abundance of certain m/z ions, some of which were identified as components of the ornbactin and pyochelin siderophores. These results highlight the chemical diversity of Burkholderia sensu lato bacteria and suggest that their capacity to inhibit the Fusarium-related infection of maize in suppressive soils is associated with siderophore synthesis.

Wide distribution of the Ustilago maydis-bacterium endosymbiosis in naturally infected maize plants.

We have previously described that laboratory strains of Ustilago maydis, a fungal pathogen of maize and its ancestor teosinte, harbor an intracellular bacterium that enables the fungus to fix nitrogen. However, it is not clear whether other strains isolated from nature also harbor endosymbiotic bacteria, and whether these fix nitrogen for its host. In the present study, we isolated U. maydis strains from naturally infected maize. All the isolated strains harbored intracellular bacteria as determined by PCR amplification of the 16S rRNA gene, and some of them showed capacity to fix nitrogen. That these are truly bacterial endosymbionts were shown by the fact that, after thorough treatments with CuSO4 followed by serial incubations with antibiotics, the aforementioned bacterial gene was still amplified in treated fungi. In all, these data support the notion that U. maydis-bacterium endosymbiosis is a general phenomenon in this species.

A novel intracellular nitrogen-fixing symbiosis made by Ustilago maydis and Bacillus spp.

We observed that the maize pathogenic fungus Ustilago maydis grew in nitrogen (N)-free media at a rate similar to that observed in media containing ammonium nitrate, suggesting that it was able to fix atmospheric N2 . Because only prokaryotic organisms have the capacity to reduce N2 , we entertained the possibility that U. maydis was associated with an intracellular bacterium. The presence of nitrogenase in the fungus was analyzed by acetylene reduction, and capacity to fix N2 by use of (15) N2 . Presence of an intracellular N2 -fixing bacterium was analyzed by PCR amplification of bacterial 16S rRNA and nifH genes, and by microscopic observations. Nitrogenase activity and (15) N incorporation into the cells proved that U. maydis fixed N2 . Light and electron microscopy, and fluorescence in situ hybridization (FISH) experiments revealed the presence of intracellular bacteria related to Bacillus pumilus, as evidenced by sequencing of the PCR-amplified fragments. These observations reveal for the first time the existence of an endosymbiotic N2 -fixing association involving a fungus and a bacterium.

Enhanced Pb Absorption by Hordeum vulgare L. and Helianthus annuus L. Plants Inoculated with an Arbuscular Mycorrhizal Fungi Consortium.

The effect of an arbuscular mycorrhizal fungi (AMF) consortium conformed by (Glomus intraradices, Glomus albidum, Glomus diaphanum, and Glomus claroideum) on plant growth and absorption of Pb, Fe, Na, Ca, and (32)P in barley (Hordeum vulgare L.) and sunflower (Helianthus annuus L.) plants was evaluated. AMF-plants and controls were grown in a substrate amended with powdered Pb slag at proportions of 0, 10, 20, and 30% v/v equivalent to total Pb contents of 117; 5,337; 13,659, and 19,913 mg Pb kg(-1) substrate, respectively. Mycorrhizal root colonization values were 70, 94, 98, and 90%, for barley and 91, 97, 95, and 97%, for sunflower. AMF inoculum had positive repercussions on plant development of both crops. Mycorrhizal barley absorbed more Pb (40.4 mg Pb kg(-1)) shoot dry weight than non-colonized controls (26.5 mg Pb kg(-1)) when treated with a high Pb slag dosage. This increase was higher in roots than shoots (650.0 and 511.5 mg Pb kg(-1) root dry weight, respectively). A similar pattern was found in sunflower. Plants with AMF absorbed equal or lower amounts of Fe, Na and Ca than controls. H. vulgare absorbed more total P (1.0%) than H. annuus (0.9%). The arbuscular mycorrizal consortium enhanced Pb extraction by plants.

Effects of iron-reducing bacteria on carbon steel corrosion induced by thermophilic sulfate-reducing consortia.

Four thermophilic bacterial species, including the iron-reducing bacterium Geobacillus sp. G2 and the sulfate-reducing bacterium Desulfotomaculum sp. SRB-M, were employed to integrate a bacterial consortium. A second consortium was integrated with the same bacteria, except for Geobacillus sp. G2. Carbon steel coupons were subjected to batch cultures of both consortia. The corrosion induced by the complete consortium was 10 times higher than that induced by the second consortium, and the ferrous ion concentration was consistently higher in iron-reducing consortia. Scanning electronic microscopy analysis of the carbon steel surface showed mineral films colonized by bacteria. The complete consortium caused profuse fracturing of the mineral film, whereas the non-iron-reducing consortium did not generate fractures. These data show that the iron-reducing activity of Geobacillus sp. G2 promotes fracturing of mineral films, thereby increasing steel corrosion.

Burkholderia cepacia XXVI siderophore with biocontrol capacity against Colletotrichum gloeosporioides.

Colletotrichum gloeosporioides is the causal agent of anthracnose in mango. Burkholderia cepacia XXVI, isolated from mango rhizosphere and identified by 16S rDNA sequencing as a member of B. cepacia complex, was more effective than 6 other mango rhizosphere bacteria in inhibiting the model mango pathogen, C. gloeosporioides ATCC MYA 456. Biocontrol of this pathogen was demonstrated on Petri-dishes containing PDA by > 90 % reduction of surface colonization. The nature of the biocontrol metabolite(s) was characterized via a variety of tests. The inhibition was almost exclusively due to production of agar-diffusible, not volatile, metabolite(s). The diffusible metabolite(s) underwent thermal degradation at 70 and 121 °C (1 atm). Tests for indole acetic acid production and lytic enzyme activities (cellulase, glucanase and chitinase) by B. cepacia XXVI were negative, indicating that these metabolites were not involved in the biocontrol effect. Based on halo formation and growth inhibition of the pathogen on the diagnostic medium, CAS-agar, as well as colorimetric tests we surmised that strain XXVI produced a hydroxamate siderophore involved in the biocontrol effect observed. The minimal inhibitory concentration test showed that 0.64 µg ml(-1) of siderophore (Deferoxamine mesylate salt-equivalent) was sufficient to achieve 91.1 % inhibition of the pathogen growth on Petri-dishes containing PDA. The biocontrol capacity against C. gloeosporioides ATCC MYA 456 correlated directly with the siderophore production by B. cepacia XXVI: the highest concentration of siderophore production in PDB on day 7, 1.7 µg ml(-1) (Deferoxamine mesylate salt-equivalent), promoted a pathogen growth inhibition of 94.9 %. The growth of 5 additional strains of C. gloeosporioides (isolated from mango "Ataulfo" orchards located in the municipality of Chahuites, State of Oaxaca in Mexico) was also inhibited when confronted with B. cepacia XXVI. Results indicate that B. cepacia XXVI or its siderophore have the potential to be used as a biological control agent against C. gloeosporioides; thus diminishing environmental problems caused by the current practices to control this disease.

Tolerance and growth of 11 Trichoderma strains to crude oil, naphthalene, phenanthrene and benzo[a]pyrene.

Petroleum hydrocarbons (PHs) are major organic contaminants in soils, whose degradation process is mediated by microorganisms such as the filamentous fungi Cunninghamella elegans and Phanerochaete chrysosporium. However, little is known about the tolerance and the degradation capability of Trichoderma species when exposed to PH. This research evaluated the tolerance and growth of 11 Trichoderma strains to crude oil (COil), naphthalene (NAPH), phenanthrene (PHE) and benzo[a]pyrene (B[a]P) by using in vitro systems. Petri dishes containing solid mineral minimum medium were separately contaminated with COil, with seven doses of either NAPH or PHE (250, 500, 750, 1000, 2000, and 3000 mg L(-1)), and with six doses of B[a]P (10, 25, 50, 75, and 100 mg L(-1)). Non-contaminated plates were used as controls. Trichoderma strains were exposed to all the contaminants by triplicate, and the growth of each fungal colony was daily recorded. No significant differences were observed among Trichoderma strains when they were exposed to COil in which the maximum fungal growth was reached at 96 h. In contrast, Trichoderma strains showed variations to tolerate and grow under different doses of either NAPH, PHE or B[a]P. Increasing NAPH doses resulted on significant greater fungal growth inhibition than PHE doses. The exposure to B[a]P did not inhibited growth of some Trichoderma strains.

Isolation and characterization of hexavalent chromium-reducing rhizospheric bacteria from a wetland.

Scirpus americanus Pers. occurs naturally in "San Germán," a pond that serves as a receptor of industrial wastewater in Guanajuato, México. This plant accumulates metals mainly in the root: concentrations (mg/kg) of Cr, As, Cd and Se were 970, 49, 41, and 85 respectively. Analysis of rhizosphere samples indicated bacterial population of 10(8) cfu g(-1) in media with 0.2 mM Cr(VI) and 10 mM sodium gluconate. Thirteen isolates were obtained and phylogenetic analyses (16S rRNA) indicated they corresponded to genera of Agrobacterium, Arthrobacter, Microbacterium, Curtobacterium, Rhodococcus, Xanthomonas and Pseudomonas. Cr(VI) reduction was evaluated using the diphenyl carbazide method. The isolates accomplished 5-40% (20 microM) of reduction in assays of resting cell and tolerated 0.5-5.0 mM Cr(VI). Eight strains used nitrate and thirteen used iron and chromium as electron acceptors to grow under anaerobic conditions. Cr(VI) reduction by five strains occurred at pH values (7-9) and NaCl concentrations (0.5-1.0 M) in basal medium. A mixed culture of strains (S17 and S28) reached a chromium removal of 100% at 0.2 mM Cr(VI) initial concentration. Aerobically, this consortium was capable of 93.8% Cr(VI) reduction of 81 microg L(-1) Cr(VI) of the industrial effluent, indicating their possible use in environmental cleanup.

Multichromosomal genome structure and confirmation of diazotrophy in novel plant-associated Burkholderia species.

Pulsed-field gel electrophoresis and 16S rRNA hybridization experiments showed that multichromosome genome structures and very large genome sizes (6.46 to 8.73 Mb) are prevalent in novel plant-associated Burkholderia species. (15)N(2) isotope dilution assays revealed unambiguous diazotrophy in these novel species. nifH gene sequence analysis, often used to determine phylogenetic relatedness among diazotrophs, showed tight clusters of Burkholderia species, which were clearly distinct from those of other diazotrophs.

Identification and characterization of sulfur-oxidizing bacteria in an artificial wetland that treats wastewater from a tannery.

Wastewater from tanneries contains high concentrations of organic matter, chromium, nitrogen, and sulfur compounds. In this study, an artificial wetland is is used as the tertiary treatment in a tannery in León Gto., México. It consists of three subplots with an area of about 450 m2. Two subplots were planted with Typha sp. and the third with Scirpus americanus. Geochemical analyses along the flowpath of the wetland show that contaminants were effectively attenuated. The most probable number technique was used to determine rhizospheric microbial populations involved in the sulfur cycle and suggested that there were 104-10(6) cells g(-1) sediment of sulfate-reducing bacteria and 10(2)-10(5) of sulfur-oxidizing bacteria (SOB). Representatives of SOB were isolated on media containing thiosulfate. Phylogenetic analysis of 16S rRNA of SOB isolates shows that they belong to the genera Acinetobacter, Alcaligenes, Ochrobactrum, and Pseudomonas. Most of the isolates are organotrophic and can oxidize reduced sulfur compounds such as elemental sulfur or thiosulfate, accumulating thiosulfate, or tetrathionate during growth. All isolates can use reduced-sulfur compounds as their sole sulfur source and some can use nitrate as an electron acceptor to grow anaerobically. Our results illustrate the relevance of SOB in the functioning of the wetland constructed for tannery wastewater remediation.

Non-Frankia actinomycetes isolated from surface-sterilized roots of Casuarina equisetifolia fix nitrogen.

Based on partial 16S sequences, we previously described a novel group of nonsymbiotic, acetylene reduction activity-positive actinomycetes which were isolated from surface-sterilized roots of Casuarina equisetifolia growing in Mexico. An amplified rRNA restriction analysis confirmed that these actinomycetes are distinct from Frankia, a finding substantiated by a 16S rRNA gene phylogenetic analysis of two of the Mexican isolates. Further support for these actinomycetes being separate from Frankia comes from the very low DNA-DNA homology that was found. Nevertheless, the Mexican isolates may be diazotrophs based not only on their ability to grow in N-free medium and reduce acetylene to ethylene but also on the results from (15)N isotope dilution analysis and the finding that a nifH gene was PCR amplified. A comparison of the nifH sequences from the various isolates showed that they are closely related to nifH from Frankia; the similarity was 84 to 98% depending on the host specificity group. An analysis of complete 16S rRNA gene sequences demonstrated that the two strains analyzed in detail are most closely related to actinobacteria in the Thermomonosporaceae and the Micromonosporaceae.

Regulatory considerations of aflatoxin contamination of food in Mexico.

Aflatoxins are potent mutagenic and carcinogenic compounds produced by some strains of Aspergillus flavus, A. nomius, and A. parasiticus that are commonly present in the environment. Human populations, particularly those whose basic diet includes grains, are in risk to exposure to aflatoxins. It is thus necessary to monitor and control the contamination of food and feed by aflatoxins in both domestic and international trade. As large amounts of corn are imported into Mexico from the United States, this paper stresses the need to develop legislation and enforce standards to ensure trade of corn with the minimal amount of aflatoxin.