Effect of Culture Temperature on 2-Methylisoborneol Production and Gene Expression in Two Strains of Pseudanabaena sp.

The presence of the odorant 2-methylisoborneol (2-MIB) in drinking water sources is undesirable. Although 2-MIB production is known to be influenced by temperature, its regulation at the gene level and its relationship with Chlorophyll-a (Chl-a) at different temperatures remain unclear. This study investigates the impact of temperature on 2-MIB production and related gene expression in Pseudanabaena strains PD34 and PD35 isolated from Lake Paldang, South Korea. The strains were cultured at three temperatures (15, 25, and 30 °C) to examine cell growth, 2-MIB production, and mic gene expression levels. 2-MIB production per cell increased with higher temperatures, whereas mic gene expression levels were higher at lower temperatures, indicating a complex regulatory mechanism involving post-transcriptional and enzyme kinetics factors. Additionally, the relationship between Chl-a and 2-MIB involved in metabolic competition was analyzed, suggesting that high temperatures appear to favor 2-MIB synthesis more than Chl-a synthesis. The distinct difference in the total amount of the two products and the proportion of 2-MIB between the two strains partially explains the variations in 2-MIB production. These findings highlight the significant effect of temperature on 2-MIB biosynthesis in Pseudanabaena and provide a valuable background for gene data-based approaches to manage issues regarding 2-MIB in aquatic environments.

Occurrence and phylogenetic analysis of Pseudanabaena sp. producing 2-methylisoborneol in drinking water source of South Korea.

We investigated the abundance of Pseudanabaena species and the concentration of the monoterpene 2-methylisoborneol (2-MIB) from July to October at three sampling sites in South Korea. To identify the main cause of 2-MIB occurrence in drinking water source, we characterized and performed a phylogenetic analysis of the 2-MIB synthase gene. Pseudanabaena was the dominant cyanobacterium (68%-100%) among the samples. At all three sampling sites, a strong positive correlation was detected between 2-MIB concentrations and Pseudanabaena cell numbers. A phylogenetic analysis of 222 MIB sequences isolated from the water samples showed that all of the clones were affiliated with the Pseudanabaena MIB synthase gene, demonstrating that the 2-MIB in the Han River drinking water source was produced by Pseudanabaena sp. Using a clone of the 2-MIB gene, network-based analysis and unweighted pair group method with arithmetic mean analysis were used to examine temporal and spatial variation in the 2-MIB concentration and Pseudanabaena abundance. The network analysis showed greater temporal than spatial similarity among the 2-MIB gene clones. Together, our results demonstrate that Pseudanabaena was the main producer of 2-MIB. These findings provide important information for odour management in drinking water source.

Low-affinity iron transport protein Uvt3277 is important for pathogenesis in the rice false smut fungus Ustilaginoidea virens.

Ustilaginoidea virens is the causal agent of rice false smut disease resulting in quantitative and qualitative losses in rice. To gain insights into the pathogenic mechanisms of U. virens, we established a T-DNA insertion mutant library of U. virens through Agrobacterium tumefaciens-mediated transformation and selected an enhanced pathogenicity mutant (i.e., B3277). We analyzed the biological characteristics of the wild-type P1 and B3277. The growth rate and sporulation of B3277 were decreased compared with those of P1; the ferrous iron could be utilized by B3277, but inhibited the growth of P1. Southern blot analysis was performed to verify the copy number of the foreign gene inserted in the genomic DNA and only one copy of the T-DNA was found. The combined hiTAIL-PCR with RACE-PCR analysis showed the successful cloning of full length of the T-DNA flanking gene associated with pathogenicity, named Uvt3277. Gene expression was analyzed using real-time PCR. Results revealed that Uvt3277 was expressed at lower levels in B3277 than in P1. This gene was then subjected to bioinformatics analysis. The encoded protein of Uvt3277 exhibited high homology with low-affinity iron transporter proteins in some fungi. Transformation of the RNAi vector by constructing the hairpin RNA of the target gene was confirmed as successful. The pathogenicity of the transformant also increased. These results suggested that Uvt3277 may have an important function associated with the pathogenesis of U. virens. This study provides insights into the pathogenic mechanism of U. virens and a molecular target of disease control.

Comparative transcriptome analysis of fruiting body and sporulating mycelia of Villosiclava virens reveals genes with putative functions in sexual reproduction.

Sexual reproduction of heterothallic clavicipitaceous fungus Villosiclava virens (anamorph: Ustilaginoidea virens) generates ascospores, which is considered as primary infection source of rice false smut disease. However, little is known about the molecular underpinnings of sexual reproduction in V. virens. In this study, transcriptomes of V. virens in fruiting body (FB) and sporulating mycelia (SM) were compared using Illumina paired-end sequencing technology. A total of 33,384,588 and 23,765,275 clean reads of FB and SM transcriptome profiles could be used to map cDNA of V. virens, respectively. We evaluated the gene expression variations between FB and SM, a total of 488 genes therein were significantly higher expressed in FB than SM, and 342 genes were significantly higher expressed genes in SM than FB. These differentially expressed genes were annotated using Kyoto Encyclopedia of Genes and Genomes and Gene Ontology databases. Several genes were found to specifically function in sexual reproduction, involving in mating type, pheromone synthesis, signaling transduction, transcription factors, and meiosis; additionally, a few of genes were presumed to function in conidia sporulation and infection. Comparative transcriptome analysis of V. virens during FB and SM provided an overview of gene expression profiles at the transcriptional level and provided hints to better understand the molecular mechanisms of sexual development. Additionally, the data presented here also proved benefit for mining of essential genes contributing to sexual conidiation and infection.

Characterization of mating-type loci in rice false smut fungus Villosiclava virens.

Ascospores of Villosiclava virens are a primary infection source of rice false smut. This phytopathogenic fungus exists in heterothallic form, and mating compatibility is regulated by mating-type locus 1 (MAT1). However, the MAT1 locus structure remains unknown. The MAT1-1 and MAT1-2 idiomorphs of V. virens were characterized and annotated on the basis of cDNA sequencing. A multiplex polymerase chain reaction (PCR) method was developed to identify the mating types of hyphae and sclerotia. The MAT1-1 locus of V. virens contains three mating-type genes: MAT1-1-1, MAT1-1-2 and MAT1-1-3, and a pseudogene similar to MAT1-2-1. The MAT1-2 locus harbors the MAT1-2-1 gene and a new mating-type gene MAT1-2-8. The mRNA of MAT1-1-1, MAT1-1-3 and MAT1-2-1, but not MAT1-1-2, was detectible by reverse transcription PCR in vegetative mycelia. However, the mRNA of MAT1-1-2 was detectible in the stroma, which is a sexual reproduction structure of V. virens. A multiplex PCR detection method was developed for the identification of the MAT1-1 and MAT1-2 idiomorphs. All 20 wild-type strains harbored either the MAT1-1 or MAT1-2 idiomorphs. Sclerotia that harbored both the MAT1-1 and MAT1-2 idiomorphs had potential to form fertile stromata, whereas those that harbored only the MAT1-1 idiomorph could not form mature stromata.

The Relationship between Depressive Symptoms in Outpatients with Chronic Illness and Health Care Costs

PURPOSE: To evaluate the relationship between depressive symptoms and health care costs in outpatients with chronic medical illnesses in Korea, we screened for depressive symptoms in 1,118 patients with a chronic medical illness and compared the severity of somatic symptoms and health care costs. PATIENTS AND METHODS: Data were compared between outpatients with depressive symptoms and those without depressive symptoms. Depression and somatic symptoms were measured by Zung's Self-rating Depression Scale (SDS) and Patient Health Questionnaire (PHQ)-15, respectively. We also investigated additional data related to patients' health care costs (number of visited clinical departments, number of visits made per patients, and health care costs). A total of 468 patients (41.9%) met the criteria for depressive disorder. RESULTS: A high rate of severe depressive symptoms was found in elderly, female and less-educated patients. A positive association between the severity of somatic symptoms and depressive symptoms was also identified. The effects of depressive symptoms in patients with chronic illnesses on three measures of health services were assessed by controlling for the effects of demographic variables and the severity of somatic symptoms. We found that the effects of depressive symptoms on the number of visited departments and number of visits made per patients were mediated by the severity of somatic symptoms. However, for health care costs, depressive symptoms had a significant main effect. Furthermore, the effect of gender on health care costs is moderated by the degree of a patient's depressive symptoms. CONCLUSION: In summary, there is clearly a need for increased recognition and treatment of depressive symptoms in outpatients with chronic medical illnesses.