The Agrobacterium tumefaciens C58 genome harbors an operon containing the dmeR (Atu0890) and dmeF (Atu0891) genes, which encode a transcriptional regulatory protein belonging to the RcnR/CsoR family and a metal efflux protein belonging to the cation diffusion facilitator (CDF) family, respectively. The dmeRF operon is specifically induced by cobalt and nickel, with cobalt being the more potent inducer. Promoter-lacZ transcriptional fusion, an electrophoretic mobility shift assay, and DNase I footprinting assays revealed that DmeR represses dmeRF transcription through direct binding to the promoter region upstream of dmeR. A strain lacking dmeF showed increased accumulation of intracellular cobalt and nickel and exhibited hypersensitivity to these metals; however, this strain displayed full virulence, comparable to that of the wild-type strain, when infecting a Nicotiana benthamiana plant model under the tested conditions. Cobalt, but not nickel, increased the expression of many iron-responsive genes and reduced the induction of the SoxR-regulated gene sodBII. Furthermore, control of iron homeostasis via RirA is important for the ability of A. tumefaciens to cope with cobalt and nickel toxicity. IMPORTANCEThe molecular mechanism of the regulation of dmeRF transcription by DmeR was demonstrated. This work provides evidence of a direct interaction of apo-DmeR with the corresponding DNA operator site in vitro. The recognition site for apo-DmeR consists of 10-bp AT-rich inverted repeats separated by six C bases (5=-ATATAGTATACCCCCCTATAGTATAT-3=). Cobalt and nickel cause DmeR to dissociate from the dmeRF promoter, which leads to expression of the metal efflux gene dmeF. This work also revealed a connection between iron homeostasis and cobalt/nickel resistance in A. tumefaciens. Cobalt is required by coenzyme B 12 -dependent enzymes and several proteins (1, 2). However, cobalt overload can cause cellular toxicity by catalyzing the generation of reactive oxygen species (3, 4), which leads to iron and glutathione depletion, and thus disturbing iron homeostasis (4-6). Cobalt competes with iron in heme proteins (7) and inhibits the activity of iron-sulfur (Fe-S) proteins as shown in Escherichia coli (5) and Salmonella enterica (6). To avoid cobalt toxicity, levels of intracellular cobalt must be properly controlled. Cobalt trafficking systems in the cell, including uptake systems, efflux systems, and metallochaperones, help maintain cobalt at levels suitable for growth (8).To prevent intracellular cobalt overload-mediated toxicity, excessive amounts of cobalt are eliminated by efflux systems involving components such as the major facilitator superfamily (MFS), P 1B-4 -ATPase, resistance nodulation cell division (RND), cation/ proton antiporter, and cation diffusion facilitator (CDF). The E. coli RcnA (resistance to cobalt and nickel) efflux pump belongs to a unique family that is responsible for the detoxification of cobalt and nickel (9). The expression of rcnA is negatively regulated by RcnR (10). CoaT is a P 1B-...

Agrobacterium tumefaciens has a cluster of genes (Atu3178, Atu3179, and Atu3180) encoding an ABC-type transporter, here named troA, troB, and troC, respectively, which is shown here to be a zinc-specific uptake system. Reverse transcription (RT)-PCR analysis confirmed that troA, troB, and troC are cotranscribed, with troC as the first gene of the operon. The yciC (Atu3181) gene is transcribed in the opposite orientation to that of the troCBA operon and belongs to a metal-binding GTPase family. Expression of troCBA and yciC was inducible under zinc-limiting conditions and was controlled by the zinc uptake regulator, Zur. Compared to the wild type, the mutant strain lacking troC was hypersensitive to a metal chelator, EDTA, and the phenotype could be rescued by the addition of zinc, while the strain with a single yciC mutation showed no phenotype. However, yciC was important for survival under zinc limitation when either troC or zinT was inactivated. The periplasmic zinc-binding protein, ZinT, could not function when TroC was inactivated, suggesting that ZinT may interact with TroCBA in zinc uptake. Unlike many other bacteria, the ABC-type transporter ZnuABC was not the major zinc uptake system in A. tumefaciens. However, the important role of A. tumefaciens ZnuABC was revealed when TroCBA was impaired. The strain containing double mutations in the znuA and troC genes exhibited a growth defect in minimal medium. A. tumefaciens requires cooperation of zinc uptake systems and zinc chaperones, including TroCBA, ZnuABC, ZinT, and YciC, for survival under a wide range of zinc-limiting conditions. IMPORTANCEBoth host and pathogen battle over access to essential metals, including zinc. In low-zinc environments, physiological responses that make it possible to acquire enough zinc are important for bacterial survival and could determine the outcome of hostpathogen interactions. A. tumefaciens was found to operate a novel pathway for zinc uptake in which ZinT functions in concert with the high-affinity zinc importer TroCBA. Zinc is an essential metal for bacteria because it is required for the functions of many enzymes and proteins (1, 2). However, zinc overload is toxic to cells (3-7). Bacteria have mechanisms to maintain zinc homeostasis via the coordinated response of genes involved in zinc uptake, efflux, and storage (8-13). The zinc uptake regulator Zur is a transcriptional regulator belonging to the Fur family and functions as a repressor of zinc uptake genes, including znuABC and zinT (10). To prevent excessive amounts of zinc in cells under high-zinc conditions, Zur uses Zn 2ϩ as its cofactor to bind to a conserved AT-rich sequence, called the Zur box (14), found in the promoter region of the zinc uptake genes, leading to inhibition of gene expression (15)(16)(17). ZnuA is a periplasmic protein that binds zinc and transfers it to the membrane permease ZnuB and the ATPase ZnuC (15). The ZinT protein is a periplasmic zinc-binding protein (18-21) that has been shown to directly interact with and assist ZnuABC i...

IntroductionAeropollen can induce detrimental effects, particularly in respiratory airways. Monitoring local aeropollen is essential for the management of pollen allergic patients in each area. However, without resources for constant monitoring, pollen counts are subjected to biases imposed by the choices of sampling season, time of collection, and location. Therefore, the effects of these factors must be better understood. This study investigated the dynamics of aeropollen types through seasonal variation, diurnal cycle and different heights from the ground in Bangkok, Thailand.MethodsAeropollen samples were collected for 12 months at the Faculty of Science, Mahidol University in Bangkok, using a RotoRod Sampler®. For the investigation of diurnal effect, pollen was collected at 7 a.m., 10 a.m., 1 p.m., 4 p.m., and 7 p.m. For the study of height effect, data were collected at 2, 10, and 18 meters above ground.Results and discussionThis is the first study of the effects of diurnal cycle and height variation on airborne pollen count in Southeast Asia. The results showed the highest concentration of aeropollen was observed in November, which was at the beginning of the northeast monsoon season in Bangkok, whereas the lowest concentration was recorded in July (rainy season). Interestingly, the lowest airborne pollen concentration recorded in July was greater than the high level of most standards. Grass pollen was found as the major aeropollen. The highest total pollen concentration was detected at 1 p.m. The maximum pollen quantity was detected at 10 meters from the ground. However, the total aeropollen concentration was extremely high (>130 grains/m3) at all elevated heights compared to other studies that mostly found at lower height (approximately 1–2 m above ground). The result suggested that pollen concentrations of most pollen types increased as height increased. This study also illustrated the correlation between aeropollen quantity and local meteorological factors.ConclusionThis aeropollen survey reported that pollen concentration and diversity were affected by seasonal variation, diurnal cycle, and height from the ground. Understanding these relationships can help with predictions of aeropollen type and quantity.

The contamination of paddy elds and rice grains by cadmium (Cd) adversely affects human health. Thus, many approaches have been proposed to reduce the accumulation of Cd in rice. Here, we investigate the potential of aqueous Moringa oleifera leaf extract (AMOLE) in decreasing uptake and toxicity of Cd in a popular Thai jasmine rice variety, Khao Dawk Mali 105 (KDML105). Plants were grown in Petri dishes, a hydroponic system, and a pot system under different concentrations of Cd, in the presence and absence of AMOLE. In Petri dishes, Cd reduced the percentage of germination by 79%, but the treatment with 0.5 mg mL -1 AMOLE signi cantly increased the germination percentage. Moreover, AMOLE signi cantly decreased Cd accumulation in rice seedlings by 97%. In the hydroponics system, 0.5 mg mL -1 AMOLE decreased Cd content in shoots by 48%. Although no signi cant physiological changes in response to Cd treatments were observed in the pot system, a large amount of Cd was accumulated in rice roots. The AMOLE treatments signi cantly reduced Cd accumulation in rice shoots and decreased Cd content in milled grain by half compared to those without AMOLE treatment. We conclude that AMOLE reduced Cd toxicity, enhanced seedling growth, and reduced Cd accumulation in rice grains.

The contamination of paddy fields and rice grains by cadmium (Cd) adversely affects human health. Thus, many approaches have been proposed to reduce the accumulation of Cd in rice. Here, we investigate the potential of aqueous Moringa oleifera leaf extract (AMOLE) in decreasing uptake and toxicity of Cd in a popular Thai jasmine rice variety, Khao Dawk Mali 105 (KDML105). Plants were grown in Petri dishes, a hydroponic system, and a pot system under different concentrations of Cd, in the presence and absence of AMOLE. In Petri dishes, Cd reduced the percentage of germination by 79%, but the treatment with 0.5 mg mL -1 AMOLE significantly increased the germination percentage. Moreover, AMOLE significantly decreased Cd accumulation in rice seedlings by 97%. In the hydroponics system, 0.5 mg mL -1 AMOLE decreased Cd content in shoots by 48%. Although no significant physiological changes in response to Cd treatments were observed in the pot system, a large amount of Cd was accumulated in rice roots. The AMOLE treatments significantly reduced Cd accumulation in rice shoots and decreased Cd content in milled grain by half compared to those without AMOLE treatment. We conclude that AMOLE reduced Cd toxicity, enhanced seedling growth, and reduced Cd accumulation in rice grains.