Enhancing cadmium phytoremediation of Chlorophytum comosum (Thunb.) Jacques by applying cadmium-resistant bacterial tablet.

This study aims to formulate bacterial tablets of cadmium (Cd)-resistant Micrococcus sp. MU1, an indole-3-acetic acid-producer, for soil inoculation to improve Cd phytoremediation by Chlorophytum comosum (Thunb.) Jacques. The viability of Micrococcus sp. MU1 in tablets after storage at room temperature and 4 °C was determined. The ability of Micrococcus sp. tablets and cell suspensions on stimulating growth and Cd accumulation in C. comosum was compared. The results found that the viability of Micrococcus sp. tablets stored at room temperature and 4 °C for 2 months were 29.2 and 97.9%, respectively. After 2 months of growth in pots, the dry biomass weights of C. comosum amended with Micrococcus sp. tablet and cell suspension were greater than that of uninoculated control by 1.4- and 1.3-fold, respectively. Cd concentrations in the roots and shoots of C. comosum inoculated with bacterial tablet and bacterial suspension were not significantly different (p < 0.05) and were greater than that of the uninoculated plants. In addition, plants inoculated with Micrococcus sp. tablet and cell suspension exhibited superior phytoextraction performance, bioaccumulation factor, and translocation factor, indicating equal performance of both bacterial forms on boosting Cd phytoremediation efficiency in C. comosum. These findings suggest that soil inoculation with Micrococcus sp. tablet as a ready-to-use inoculum is a novel approach to promote phytoremediation of C. comosum in Cd-contaminated agricultural soil.

Seasonal variation in the abundance of microplastics in three commercial bivalves from Bandon Bay, Gulf of Thailand.

This study investigated the abundance of microplastics in three commercial bivalves found at Bandon Bay. Spatial-temporal differences in the concentration of microplastics were evaluated during the dry and wet seasons. The results showed that the highest abundance of microplastics in oysters, blood cockles, and green mussels was observed in fishery and aquaculture areas during the dry season, with 1.42 particles/g (w/w), 1.01 particles/g (w/w) and 0.87 particles/g (w/w), respectively. Microplastics were more abundant during the dry season compared to the wet season (p < 0.05), with fibre being the predominant shape and black being the major colour of particles. Cellophane was the most common type of polymer in all bivalves. This result is an important reference for understanding the status of microplastics in three commercial bivalves during different seasons and in different human activities, which should aid in understanding the sources of microplastics in Bandon Bay.

Cadmium-resistant Streptomyces stimulates phytoextraction potential of Crotalaria juncea L. in cadmium-polluted soil.

This work evaluated the competence of two strains of cadmium (Cd)-resistant Streptomyces, namely Streptomyces rapamycinicus K5PN1, an indole-3-acetic acid (IAA) producer, and Streptomyces cyaneus 11-10SHTh, a siderophore producer on promoting Cd phytoextraction by sunn hemp. The results showed that S. rapamycinicus improved root elongation of sunn hemp seedlings under Cd stress conditions. S. rapamycinicus and S. cyaneus were colonized on the root surface of sunn hemp at concentrations of 2.3 × 104 and 6.4 × 103 CFU g-1 root fresh weight, respectively. The results of pot-culture experiments showed that S. rapamycinicus increased the root and shoot lengths, and dry biomass of sunn hemp planted in high Cd-contaminated soil. The Cd concentration in the leaves of sunn hemp inoculated with S. cyaneus (73.82 ± 2.20 mg kg-1 plant dry wt) was higher than that of plants with S. rapamycinicus inoculation and the uninoculated control. The phytoextraction of Cd by sunn hemp was significantly increased with Cd-resistant Streptomyces inoculation. In conclusion, both strains of Cd-resistant Streptomyces had potential on enhancing Cd phytoextraction efficiency of sunn hemp. Our study suggests the application of Cd-resistant Streptomyces can improve Cd phytoextraction by sunn hemp for restoration of Cd-polluted sites.

Our study demonstrated the potential of two strains of Cd-resistant Streptomyces to stimulate Cd phytoextraction from soil by sunn hemp. Cadmium-resistant Streptomyces strongly stimulated Cd uptake and accumulation in sunn hemp planted in high level of Cd-polluted soil, supporting sunn hemp to be a superior Cd accumulator.

Microplastics in the surface seawater of Bandon Bay, Gulf of Thailand.

This study aimed to evaluate the microplastics abundance, composition and distribution in Bandon Bay's surface seawater, in southern Thailand. Samples of microplastics were collected from 48 transects using a surface manta trawl at four different estuaries that support human activities. The results showed that the highest microplastic abundance occurred in the fishery and aquaculture areas with a mean abundance of 0.33 particles/m3. Fragments were the dominant form at all stations. Microplastics with <1 mm were the dominant size, and white was the colour most found in all stations. Polypropylene was the major type of microplastic, accounting for 57% overall. This study is an important reference for understanding the microplastics status in the surface seawater of Bandon Bay, as it will allow relevant agencies to accurately assess the pollution level of microplastics in the bay. It is of practical significance to understand the sources and sinks of microplastics.

Influence of cadmium-resistant Streptomycetes on plant growth and cadmium uptake by Chlorophytum comosum (Thunb.) Jacques.

This work aims to explore the role of cadmium-resistant actinomycetes on promoting plant growth and cadmium uptake in Chlorophytum comosum (Thunb.) Jacques, a spider plant. Actinomycetes isolated from the plant roots in peat swamp forests were screened for their cadmium resistance and the production of indole-3-acetic acid (IAA) and siderophores. The results found that K5PN1 and 11-10SHTh produced high levels of IAA and siderophores, respectively. K5PN1 and 11-10SHTh were identified to be Streptomyces rapamycinicus and Streptomyces cyaneus, respectively. Both strains were able to remove cadmium from aqueous solution and survive under cadmium stress in contaminated soil. The results of pot experiments found that the selected Streptomyces inoculation increased the root and shoot biomass and cadmium accumulation in the root and shoot of C. comosum planted in a cadmium-contaminated soil. The highest cadmium accumulation and translocation ability of cadmium from the root to shoot was found in C. comosum with S. rapamycinicus inoculation. In addition, plant with S. cyaneus inoculation had the highest phytoextraction coefficient and bioaccumulation factor. Our findings concluded that S. rapamycinicus and S. cyaneus stimulated the growth and cadmium uptake in C. comosum, suggesting a combined approach using the selected Streptomyces and C. comosum for phytoremediation of cadmium-polluted soil.

Effects of biochar-immobilized bacteria on phytoremediation of cadmium-polluted soil.

This work is the first report of the ability of biochar-immobilized cadmium-resistant bacteria (CRB) on promoting the efficiency of cadmium phytoextraction by Chlorophytum laxum R.Br. The survival of CRB immobilized on biochar in cadmium-contaminated soil at a concentration of 75.45 mg kg-1 was studied. The results found that both CRB, namely Arthrobacter sp. TM6 and Micrococcus sp. MU1, can survive and grow in cadmium-contaminated soil. To study phytoextraction in the pot experiments, 2-month-old C. laxum was individually planted in cadmium-contaminated soil and divided into four treatments, including (i) untreated control, (ii) biochar, (iii) biochar-immobilized (BC) Arthrobacter sp., and (iv) BC-Micrococcus sp. The results found that biochar-immobilized CRB did not cause any effect to the root lengths and shoot heights of plants compared to the untreated control. Interestingly, inoculation of biochar-immobilized CRB significantly increased cadmium accumulation in the shoots and roots compared to the untreated control. In addition, the highest cadmium content in a whole plant, best phytoextraction performance, and greatest bioaccumulation factor was found in plant inoculated with BC-Micrococcus sp., followed by BC-Arthrobacter sp. In conclusion, inoculation of biochar-immobilized CRB enhanced cadmium accumulation and translocation of cadmium from the roots to shoots, suggesting further applying biochar-immobilized CRB in cadmium-polluted soil for promoting cadmium phytoextraction efficiency of ornamental plants. Graphical abstract.

Potential of Napier grass with cadmium-resistant bacterial inoculation on cadmium phytoremediation and its possibility to use as biomass fuel.

This work mainly aims to explore the potential of synergistic use of cadmium-resistant bacteria and Napier grass to promote cadmium phytoremediation and the possibility of using the harvested Napier grass for biomass fuel. A pot experiment was carried out by transplanting Napier grass with and without bacterial inoculation in cadmium contaminated soil for 6 months. The results found that Micrococcus sp. significantly promoted the shoot biomass of Napier grass but not the root biomass. Micrococcus sp. and Arthrobacter sp. stimulated cadmium accumulation in the root and the shoot. Cadmium was retained more in the root than the shoot at all plantation periods. The maximum cadmium content in a whole plant was found in plants inoculated with Micrococcus sp. at six months. The values of phytoextraction coefficient and bioaccumulation factor in plants with bacterial inoculation were higher than those in the uninoculated control. Translocation factor was very low. Napier grass could be considered as a candidate plant for cadmium phytostabilization. The calorific value of Napier grass transplanted in cadmium-contaminated soil was similar to that in uncontaminated soil, but cadmium was still retained in the ash and some was emitted into the air. In conclusion, these cadmium-resistant bacteria enhanced the performance of Napier grass on cadmium phytoremediation. The harvested Napier grass can be used for biomass fuel under controlled ash and air emission from the combustion process.

Phytoremediation of cadmium-polluted soil by Chlorophytum laxum combined with chitosan-immobilized cadmium-resistant bacteria.

This study examined the performance of the chitosan-immobilized cadmium-resistant bacteria Arthrobacter sp. and Micrococcus sp. on cadmium phytoremediation by Chlorophytum laxum in cadmium-polluted soil. These immobilized cadmium-resistant bacteria can survive in cadmium-contaminated soil and significantly increased soil cadmium solubility, but the ability of chitosan-immobilized cells to increase cadmium solubility was lower than that of free cells. A pot experiment demonstrated that chitosan-immobilized Micrococcus sp. promoted the growth of C. laxum planted in cadmium-contaminated soil. A significant increase in the cadmium concentration in the roots and aboveground parts of C. laxum was found in plants inoculated with free and chitosan-immobilized cells of these bacteria. The performance of Arthrobacter sp. free cells to augment cadmium accumulation in C. laxum was a little bit better than that of chitosan-immobilized Arthrobacter sp., except at 9 weeks after planting. The phytoextraction coefficient, bioaccumulation factor, and translocation factor of C. laxum inoculated with free and chitosan-immobilized cells of cadmium-resistant bacteria were higher than those of the uninoculated control and increased with time. Our findings suggest that chitosan-immobilized cells can be exploited to enhance the efficiency of cadmium phytoremediation by C. laxum.

Enhanced cadmium phytoremediation of Glycine max L. through bioaugmentation of cadmium-resistant bacteria assisted by biostimulation.

This study examined the potential of three strains of cadmium-resistant bacteria, including Micrococcus sp., Pseudomonas sp. and Arthrobacter sp., to promote root elongation of Glycine max L. seedlings, soil cadmium solubility and cadmium phytoremediation in G. max L. planted in soil highly polluted with cadmium with and without nutrient biostimulation. Micrococcus sp. promoted root length in G. max L. seedlings under toxic cadmium conditions. Soil inoculation with Arthrobacter sp. increased the bioavailable fraction of soil cadmium, particularly in soil amended with a C:N ratio of 20:1. Pot culture experiments observed that the highest plant growth was in Micrococcus sp.-inoculated plants with nutrient biostimulation. Cadmium accumulation in the roots, stems and leaves of G. max L. was significantly enhanced by Arthrobacter sp. with nutrient biostimulation. A combined use of G. max L. and Arthrobacter sp. with nutrient biostimulation accelerated cadmium phytoremediation. In addition, cadmium was retained in roots more than in stems and leaves and G. max L. had the lowest translocation factor at all growth stages, suggesting that G. max L. is a phytostabilizing plant. We concluded that biostimulation-assisted bioaugmentation is an important strategy for improving cadmium phytoremediation efficiency.

Improvement of cadmium phytoremediation after soil inoculation with a cadmium-resistant Micrococcus sp.

Cadmium-resistant Micrococcus sp. TISTR2221, a plant growth-promoting bacterium, has stimulatory effects on the root lengths of Zea mays L. seedlings under toxic cadmium conditions compared to uninoculated seedlings. The performance of Micrococcus sp. TISTR2221 on promoting growth and cadmium accumulation in Z. mays L. was investigated in a pot experiment. The results indicated that Micrococcus sp. TISTR2221significantly promoted the root length, shoot length, and dry biomass of Z. mays L. transplanted in both uncontaminated and cadmium-contaminated soils. Micrococcus sp. TISTR2221 significantly increased cadmium accumulation in the roots and shoots of Z. mays L. compared to uninoculated plants. At the beginning of the planting period, cadmium accumulated mainly in the shoots. With a prolonged duration of cultivation, cadmium content increased in the roots. As expected, little cadmium was found in maize grains. Soil cadmium was significantly reduced with time, and the highest percentage of cadmium removal was found in the bacterial-inoculated Z. mays L. after transplantation for 6 weeks. We conclude that Micrococcus sp. TISTR2221 is a potent bioaugmenting agent, facilitating cadmium phytoextraction in Z. mays L.

Bioaugmentation with cadmium-resistant plant growth-promoting rhizobacteria to assist cadmium phytoextraction by Helianthus annuus.

Micrococcus sp. MU1 and Klebsiella sp. BAM1, the cadmium-resistant plant growth-promoting rhizobacteria (PGPR), produce high levels of indole-3-acetic acid (IAA) during the late stationary phase of their growth. The ability of PGPR to promote root elongation, plant growth and cadmium uptake in sunflowers (Helianthus annuus) was evaluated. Both species of bacteria were able to remove cadmium ions from an aqueous solution and enhanced cadmium mobilization in contaminated soil. Micrococcus sp. and Klebsiella sp. use aminocyclopropane carboxylic acid as a nitrogen source to support their growth, and the minimum inhibitory concentrations of cadmium for Micrococcus sp. and Klebsiella sp. were 1000 and 800mM, respectively. These bacteria promoted root elongation in H. annuus seedlings in both the absence and presence of cadmium compared to uninoculated seedlings. Inoculation with these bacteria was found to increase the root lengths of H. annuus that had been planted in cadmium-contaminated soil. An increase in dry weight was observed for H. annuus inoculated with Micrococcus sp. Moreover, Micrococcus sp. enhanced the accumulation of cadmium in the root and leaf of H. annuus compared to untreated plants. The highest cadmium accumulation in the whole plant was observed when the plants were treated with EDTA following the treatment with Micrococcus sp. In addition, the highest translocation of cadmium from root to the above-ground tissues of H. annuus was found after treatment with Klebsiella sp. in the fourth week after planting. Our results show that plant growth and cadmium accumulation in H. annuus was significantly enhanced by cadmium-resistant PGPRs, and these bacterial inoculants are excellent promoters of phytoextraction for the rehabilitation of heavy metal-polluted environments.

Bacillus subtilis SSE4 produces subtulene A, a new lipopeptide antibiotic possessing an unusual C15 unsaturated beta-amino acid.

Subtulene A, a new cyclic lipopeptide, was isolated from the culture broth of Bacillus subtilis SSE4. This antibiotic compound contained the seven common alpha-amino acids, L-Asn-1, D-Tyr-2, D-Asn-3, L-Gln-4, L-Pro-5, D-Asn-6, L-Ser-7 and the unique beta-amino acid-8 present in the iturin family. 1D and 2D NMR, as well as MS analyses, identified the beta-amino acid as 3-amino-13-methyltetradec-8-enoic acid, an Iso C15 long chain beta-amino acid. B. subtilis SSE4 was also found to produce iturin A. B. subtilis SSE4 culture filtrate exhibited both antifungal and antibacterial activities.

Adaptive and cross-protective responses against cadmium and zinc toxicity in cadmium-resistant bacterium isolated from a zinc mine

Cadmium (Cd) is a major environmental hazard, which usually is detected in its ionic form of Cd2+. It also causes adverse toxic effects on human health and other living organisms. Cd-resistant bacteria were isolated from Cd-contaminated soils. One isolate, TAK1, was highly resistance level to Cd toxicity. TAK1 was isolated from soil contaminated with a high Cd concentration (204.1 mg.kg-1). The result of 16S rDNA sequence analysis found that the TAK1 showed the similarity to Ralstonia sp. Physiological adaptive and cross-protective responses to Cd and Zn killing were investigated in Ralstonia sp.TAK1. Exposure to a low concentration of Cd induced adaptive resistance to higher concentrations of Cd. In addition, pretreatment of Ralstonia sp.TAK1 with an inducing concentration of Cd conferred cross-protective response against subsequent exposure to the lethal concentrations of Zn. The induced adaptive and cross-protective response Ralstonia sp.TAK1 required newly synthesized protein(s). Cd-induced adaptive and cross-protective responses against Cd and Zn toxicity are the important mechanisms used by Ralstonia sp.TAK1 to survive in the heavy metal contaminated environments. These findings might lead to the use of Ralstonia sp.TAK1 for microbial based remediation in Cd and Zn-contaminated soils.

Adaptive and cross-protective responses against cadmium and zinc toxicity in cadmium-resistant bacterium isolated from a zinc mine.

Cadmium (Cd) is a major environmental hazard, which usually is detected in its ionic form of Cd(2+). It also causes adverse toxic effects on human health and other living organisms. Cd-resistant bacteria were isolated from Cd-contaminated soils. One isolate, TAK1, was highly resistance level to Cd toxicity. TAK1 was isolated from soil contaminated with a high Cd concentration (204.1 mg.kg(-1)). The result of 16S rDNA sequence analysis found that the TAK1 showed the similarity to Ralstonia sp. Physiological adaptive and cross-protective responses to Cd and Zn killing were investigated in Ralstonia sp.TAK1. Exposure to a low concentration of Cd induced adaptive resistance to higher concentrations of Cd. In addition, pretreatment of Ralstonia sp.TAK1 with an inducing concentration of Cd conferred cross-protective response against subsequent exposure to the lethal concentrations of Zn. The induced adaptive and cross-protective response Ralstonia sp.TAK1 required newly synthesized protein(s). Cd-induced adaptive and cross-protective responses against Cd and Zn toxicity are the important mechanisms used by Ralstonia sp.TAK1 to survive in the heavy metal-contaminated environments. These findings might lead to the use of Ralstonia sp.TAK1 for microbial based remediation in Cd and Zn-contaminated soils.

Antifungal potential of extracellular metabolites produced by Streptomyces hygroscopicus against phytopathogenic fungi.

Indigenous actinomycetes isolated from rhizosphere soils were assessed for in vitro antagonism against Colletotrichum gloeosporioides and Sclerotium rolfsii. A potent antagonist against both plant pathogenic fungi, designated SRA14, was selected and identified as Streptomyces hygroscopicus. The strain SRA14 highly produced extracellular chitinase and beta-1,3-glucanase during the exponential and late exponential phases, respectively. Culture filtrates collected from the exponential and stationary phases inhibited the growth of both the fungi tested, indicating that growth suppression was due to extracellular antifungal metabolites present in culture filtrates. The percentage of growth inhibition by the stationary culture filtrate was significantly higher than that of exponential culture filtrate. Morphological changes such as hyphal swelling and abnormal shapes were observed in fungi grown on potato dextrose agar that contained the culture filtrates. However, the antifungal activity of exponential culture filtrates against both the experimental fungi was significantly reduced after boiling or treatment with proteinase K. There was no significant decrease in the percentage of fungal growth inhibition by the stationary culture filtrate that was treated as above. These data indicated that the antifungal potential of the exponential culture filtrate was mainly due to the presence of extracellular chitinase enzyme, whereas the antifungal activity of the stationary culture filtrate involved the action of unknown thermostable antifungal compound(s).

The role of chitosan in protection of soybean from sudden death syndrome caused by Fusarium solani f. sp. glycines.

The in vitro antifungal properties of chitosan and its role in protection of soybean from a sudden death syndrome (SDS) were evaluated. Chitosan inhibited the radial and submerged growth of F. solani f. sp. glycines with a marked effect at concentrations up to 1mg/ml indicating antifungal property and at 3mg/ml was able to delay SDS symptoms expression on soybean leaves for over three days after fungal inoculation when applied preventively. Chitosan was able to induce the level of chitinase activity in soybean resulting in the retardation of SDS development in soybean leaves. However, the SDS symptoms gradually appeared and were associated with the reduction of chitinase activity level after five days of infection period. These results suggested the role of chitosan in partially protecting soybeans from F. solani f. sp. glycines infection.

Challenging Xanthomonas campestris with low levels of arsenic mediates cross-protection against oxidant killing.

Xanthomonas encounters highly toxic reactive oxygen species (ROS) from many sources, such as those generated by plants against invading bacteria, other soil bacteria and from aerobic respiration. Thus, conditions that alter intracellular ROS levels such as exposure to toxic metalloids would have profound effects on bacterial physiology. Here, we report that exposure of Xanthomonas campestris pv. phaseoli (Xp) to low levels of arsenic induces physiological cross-protection against killing by H(2)O(2) and organic hydroperoxide but not a superoxide generator. Cross-protection against H(2)O(2) and organic hydroperoxide toxicity was due to increased expression of genes encoding major peroxide-metabolizing enzymes such as alkyl hydroperoxide reductase (AhpC), catalase (KatA) and organic hydroperoxide resistance protein (Ohr). Arsenic-induced protection against H(2)O(2) and organic hydroperoxide requires the peroxide stress response regulators, OxyR and OhrR, respectively. Moreover, analyses of double mutants of the major H(2)O(2) and organic hyproperoxide-scavenging enzymes, Xp ahpC katA and Xp ahpC ohr, respectively, suggested the existence of unidentified OxyR- and OhrR-regulated genes that are involved in arsenic-induced resistance to H(2)O(2) and organic hyproperoxide killing in Xp. These arsenic-induced physiological alterations could play an important role in bacterial survival both in the soil environment and during plant-pathogen interactions.

ohrR and ohr are the primary sensor/regulator and protective genes against organic hydroperoxide stress in Agrobacterium tumefaciens.

The genes involved in organic hydroperoxide protection in Agrobacterium tumefaciens were functionally evaluated. Gene inactivation studies and functional analyses have identified ohr, encoding a thiol peroxidase, as the gene primarily responsible for organic hydroperoxide protection in A. tumefaciens. An ohr mutant was sensitive to organic hydroperoxide killing and had a reduced capacity to metabolize organic hydroperoxides. ohr is located next to, and is divergently transcribed from, ohrR, encoding a sensor and transcription regulator of organic hydroperoxide stress. Transcription of both ohr and ohrR was induced by exposure to organic hydroperoxides but not by exposure to other oxidants. This induction required functional ohrR. The results of gel mobility shift and DNase I footprinting assays with purified OhrR, combined with in vivo promoter deletion analyses, confirmed that OhrR regulated both ohrR and ohr by binding to a single OhrR binding box that overlapped the ohrR and ohr promoters. ohrR and ohr are both required for the establishment of a novel cumene hydroperoxide-induced adaptive response. Inactivation or overexpression of other Prx family genes (prx1, prx2, prx3, bcp1, and bcp2) did not affect either the resistance to, or the ability to degrade, organic hydroperoxide. Taken together, the results of biochemical, gene regulation and physiological studies support the role of ohrR and ohr as the primary system in sensing and protecting A. tumefaciens from organic hydroperoxide stress.

Protection of Xanthomonas against arsenic toxicity involves the peroxide-sensing transcription regulator OxyR.

Arsenic has been shown to mediate its toxicity through induced generation of reactive oxygen species. Here, we examined the role of oxidative stress-inducible genes (katA, ahpC and ohr) and their regulators (oxyR and ohrR) in the response to arsenic treatment in a plant pathogenic bacterium, Xanthomonas campestris pv. phaseoli (Xp). Overproduction of peroxide-scavenging enzymes (KatA, AhpCF and Ohr) did not enhance arsenic tolerance in wild-type Xp. Furthermore, inactivation of katA, ahpC, ohr, and ohrR genes had no effect on the level of arsenic resistance. By contrast, an oxyR mutant (Xp oxyR) showed increased sensitivity to both pentavalent arsenate and, to a greater extent, trivalent arsenite. The resistance of cells to arsenite treatment was significantly affected by the level of iron. Cells were 10-fold more sensitive to arsenite killing in the presence of excess iron, while removal of iron by an iron chelator (2,2'-dipyridyl) protected Xanthomonas from arsenite toxicity. The arsenite-sensitive phenotype of Xp oxyR could be complemented by the expression of functional OxyR from a plasmid vector, but not by the expression of other known OxyR-regulated peroxide-scavenging enzymes such as KatA and AhpCF, Ohr and OhrR. The data suggested that as yet unidentified, OxyR-regulated gene(s) are involved in conferring arsenic resistance in Xp. To our knowledge, this is the first report showing that the peroxide-sensing regulator OxyR is involved in arsenic resistance.

Analysis of growth phase regulated KatA and CatE and their physiological roles in determining hydrogen peroxide resistance in Agrobacterium tumefaciens.

Agrobacterium tumefaciens possesses two catalases, a bifunctional catalase-peroxidase, KatA and a homologue of a growth phase regulated monofunctional catalase, CatE. In stationary phase cultures and in cultures entering stationary phase, total catalase activity increased 2-fold while peroxidase activity declined. katA and catE were found to be independently regulated in a growth phase dependent manner. KatA levels were highest during exponential phase and declined as cells entered stationary phase, while CatE was detectable at early exponential phase and increased during stationary phase. Only small increases in H2O2 resistance levels were detected as cells entering stationary phase. The katA mutant was more sensitive to H2O2 than the parental strain during both exponential and stationary phase. Inactivation of catE alone did not significantly change the level of H2O2 resistance. However, the katA catE double mutant was more sensitive to H2O2 during both exponential and stationary phase than either of the single catalase mutants. The data indicated that KatA plays the primary role and CatE acts synergistically in protecting A. tumefaciens from H2O2 toxicity during all phases of growth. Catalase-peroxidase activity (KatA) was required for full H2O2 resistance. The expression patterns of the two catalases in A. tumefaciens reflect their physiological roles in the protection against H2O2 toxicity, which are different from other bacteria.