Integrated genome-transcriptome analysis unveiled the mechanism of Debaryomyces hansenii-mediated arsenic stress amelioration in rice.

Globally, rice is becoming more vulnerable to arsenic (As) pollution, posing a serious threat to public food safety. Previously Debaryomyces hansenii was found to reduce grain As content of rice. To better understand the underlying mechanism, we performed a genome analysis to identify the key genes in D. hansenii responsible for As tolerance and plant growth promotion. Notably, genes related to As resistance (ARR, Ycf1, and Yap) were observed in the genome of D. hansenii. The presence of auxin pathway and glutathione metabolism-related genes may explain the plant growth-promoting potential and As tolerance mechanism of this novel yeast strain. The genome annotation of D. hansenii indicated that it contains a repertoire of genes encoding antioxidants, well corroborated with the in vitro studies of GST, GR, and glutathione content. In addition, the effect of D. hansenii on gene expression profiling of rice plants under As stress was also examined. The Kyoto Encyclopedia of Genes and Genomes (KEGG) database revealed 307 genes, annotated in D. hansenii-treated rice, related to metabolic pathways (184), photosynthesis (12), glutathione (10), tryptophan (4), and biosynthesis of secondary metabolite (117). Higher expression of regulatory elements like AUX/IAA and WRKY transcription factors (TFs), and defense-responsive genes dismutases, catalases, peroxiredoxin, and glutaredoxins during D. hansenii+As exposure was also observed. Combined analysis revealed that D. hansenii genes are contributing to stress mitigation in rice by supporting plant growth and As-tolerance. The study lays the foundation to develop yeast as a beneficial biofertilizer for As-prone areas.

Mitigation of arsenic toxicity in rice by the co-inoculation of arsenate reducer yeast with multifunctional arsenite oxidizing bacteria.

The study aimed to explicate the role of microbial co-inoculants for the mitigation of arsenic (As) toxicity in rice. Arsenate (AsV) reducer yeast Debaryomyces hansenii NBRI-Sh2.11 (Sh2.11) with bacterial strains of different biotransformation potential was attempted to develop microbial co-inoculants. An experiment to test their efficacy (yeast and bacterial strains) on plant growth and As uptake was conducted under a stressed condition of 20 mg kg-1 of arsenite (AsIII). A combination of Sh2.11 with an As(III)-oxidizer, Citrobacter sp. NBRI-B5.12 (B5.12), resulted in ∼90% decrease in grain As content as compared to Sh2.11 alone (∼40%). Reduced As accumulation in rice roots under co-treated condition was validated with SEM-EDS analysis. Enhanced As expulsion in the selected combination under in vitro conditions was found to be correlated with higher As content in the soil during their interaction with plants. Selected co-inoculant mediated enhanced nutrient uptake in association with better production of indole acetic acid (IAA) and gibberellic acid (GA) in shoot, support microbial co-inoculant mediated better biomass under stressful condition. Boosted defense response in association with enhanced glutathione-S-transferase (GST) and glutathione reductase (GR), activities under in vitro and in vivo conditions were observed. These results indicated that the As(III) oxidizer-B5.12 accelerated the As detoxification property of the As(V) reducer-Sh2.11. Henceforth, the results confer that the coupled reduction-oxidation process of the co-inoculant reduces the accumulation of As in rice grain. These co-inoculants can be further developed for field trials to achieve higher biomass with alleviated As toxicity in rice.

Production of flavor compounds from rice bran by yeasts metabolisms of Kluyveromyces marxianus and Debaryomyces hansenii.

The aim of this study was to evaluate the biosynthesis of flavor compounds from rice bran by fermentation facilitated by Kluyveromyces marxianus and Debaryomyces hansenii. The growth of both yeasts was assessed by specific growth rates and doubling time. The biosynthesis of flavor compounds was evaluated by gas chromatography-olfactometry (GC-O), gas chromatography-mass spectrometry (GC-MS), and Spectrum™ sensory analysis. The specific growth rate (µ) and doubling time (td) of K. marxianus was calculated as 0.16/h and 4.21h, respectively, whereas that of D. hansenii was determined as 0.13/h and 5.33h, respectively. K. marxianus and D. hansenii produced significant levels of higher alcohols and acetate esters from rice bran. Results showed that K. marxianus can produce 827.27 µg/kg of isoamyl alcohol, 169.77 µg/kg of phenyl ethyl alcohol, and 216.08 µg/kg of phenyl ethyl acetate after 24-h batch fermentation. A significant amount of isovaleric acid was also synthesized by K. marxianus (4013 µg/kg) after the batch fermentation of 96 h. 415.64 µg/kg of isoamyl alcohol and 135.77 µg/kg of phenyl ethyl acetate was determined in rice bran fermented by D. hansenii after 24-h fermentation. Fermented cereals and rose were the characteristic flavor descriptors of the fermented rice bran samples. Rose flavor in fermented rice bran samples was found to be associated with phenyl ethyl alcohol, phenyl ethyl acetate, isoamyl acetate, and guaiacol. Thus, the findings of this study demonstrate that the valorization of rice bran can be achieved with the production of natural flavor compounds by yeast metabolism.

Assessment of endophytic yeast diversity in rice leaves by a culture-independent approach.

Endophytic microorganisms inhabit internal plant tissues in the host plant without causing any symptoms or negative effects. Although the diversity of endophytes has been evaluated by both culture-dependent and culture-independent methods, less information is available on yeast communities. Therefore, in this study a culture-independent method was used to examine endophytic yeasts associated with rice leaves based on the large subunit of ribosomal DNA using a semi-nested PCR technique. Sequence analysis indicated that the colonization frequency and the relative species frequency (RF) of endophytic yeast phylotypes were 0.41 and 0.06, respectively, and the majority of the yeast phylotypes were basidiomycetous yeasts. The phylotypes were designated as five known species (Cryptococcus victoriae, Debaryomyces hansenii, Debaryomyces vindobonensis, Meyerozyma guilliermondii and Pseudozyma antarctica), together with seventeen phylotypes closest to Candida metapsilosis, Cryp. foliicola, Cryp. laurentii, Pseudozyma abaconensis, Pseudozyma aphidis and Trichosporon asahii, among which some could be novel species. The most prevalent phylotypes were those closest to Cryp. foliicola (47.5 % RF) followed by D. hansenii (22.8 % RF) and P. antarctica (16.8 % RF). The presence of the phylotypes related to species known for their potential applications as biocontrol agents and plant growth promoting hormone producers suggests that they may have valuable applications. In addition, our findings revealed the occurrence of novel phylotypes at high frequency, which should encourage extensive studies to discover novel yeast species and to understand their roles in the rice leaves.

Calidad bacteriológica y detección de Bacillus cereus toxigénicos en arroz blanco cocido expendido en el área metropolitana de la provincia de San José, Costa Rica

El gran consumo de arroz a nivel mundial es uno de los factores que favorece su implicación en brotes de origen alimentario y de uno de los patógenos más importantes ligado a este producto como el Bacillus cereus El objetivo del presente trabajo fue evaluar la calidad microbiológica de 50 muestras de arroz blanco cocido expendido en restaurantes de área Metropolitana de San José Costa Rica, incluyendo la determinación del recuento total aerobio mesófilo, Número Más Probable de coliformes totales, fecales y E. coli, B. cereus así como de detección de sus genes nheA, nheB, y nheC. Para el análisis bacteriológico se siguieron los procedimientos descritos en el Compendio de Métodos para el Examen Microbiológico de Alimentos y para la detección de los genes se utililzó un PCR múltiplex y la metodología descrita por Hansen et al., 2001. De las muestras analizadas 46% fueron positivas por coliformes totales, 34% por coliformes fecales, 16% por E. coli, 10% por B. cereus y un 8% por B. cereus toxigénico Lo anterior sugiere que el consumo de arroz blanco en restaurantes puede representar un riesgo para la salud pública y que es necesario implementar mejoras con el fin de brindarle al consumidor un producto inocuo y de mejor calidad.

Bacteriological quality and toxigenic Bacillus cereus detection in cooked white rice sold at the Metropolitan Area of San José, Costa Rica.. The wide use of rice is one of the factors that favors its implication in food borne diseases, and one of the most important pathogens associated to it is Bacillus cereus. The aim of this work was to evaluate the microbiological quality of 50 samples of white cooked rice sold in restaurants at the Metropolitan Area of San José, Costa Rica, including the determination of the total aerobic plate count, the Most Probable Number of total and fecal coliforms and Escherichia coli. MPN of Bacillus cereus and the detection of nheA, nheB and nHeC genes, associated to its toxicity, was also performed. Procedures described in the Compendium of Methods for the Microbiological Examination of Foods were followed for the bacteriological analysis, multiplex PCR was used for the detection of genes following the methodology described by Hansen et al, 2001. 46% of the samples analysed were positive for total coliforms, 34% for fecal coliforms, 16% for E. coli and 10% for B. cereus, being 8% toxigenic. These facts suggest that white cooked rice may represent a risk for Pubic Health and that improvements shall be performed in order to offer a safe and high quality product to consumers.

[Bacteriological quality and toxigenic Bacillus cereus detection in cooked white rice sold at the metropolitan area of San José, Costa Rica]. / Calidad bacteriológica y detección de Bacillus cereus toxigénicos en arroz blanco cocido expendido en el área metropolitana de la provincia de San José, Costa Rica.

The wide use of rice is one of the factors that favors its implication in food borne diseases, and one of the most important pathogens associated to it is Bacillus cereus. The aim of this work was to evaluate the microbiological quality of 50 samples of white cooked rice sold in restaurants at the Metropolitan Area of San José, Costa Rica, including the determination of the total aerobic plate count, the Most Probable Number of total and fecal coliforms and Escherichia coli. MPN of Bacillus cereus and the detection of nheA, nheB and nHeC genes, associated to its toxicity, was also performed. Procedures described in the Compendium of Methods for the Microbiological Examination of Foods were followed for the bacteriological analysis, multiplex PCR was used for the detection of genes following the methodology described by Hansen et al, 2001. 46% of the samples analysed were positive for total coliforms, 34% for fecal coliforms, 16% for E. coli and 10% for B. cereus, being 8% toxigenic. These facts suggest that white cooked rice may represent a risk for Pubic Health and that improvements shall be performed in order to offer a safe and high quality product to consumers.