Thiosulfate Oxidation and mixotrophic growth of Methylobacterium goesingense and Methylobacterium fujisawaense.

The mixotrophic growth with methanol plus thiosulfate was examined in nutrient-limited mixotrophic condition for Methylobacterium goesingense CBMB5 and Methylobacterium fujisawaense CBMB37. Thiosulfate oxidation increased the growth and protein yield in mixotrophic medium that contained 150 mM methanol and 20 mM sodium thiosulfate, at 144 h. Respirometric study revealed that thiosulfate was the most preferable reduced inorganic sulfur source, followed by sulfite and sulfur. M. goesingense CBMB5 and M. fujisawaense CBMB37 oxidized thiosulfate directly to sulfate, and intermediate products of thiosulfate oxidation such as polythionates, sulfite, and sulfur were not detected in spent medium and they did not yield positive amplification for tested soxB primers. Enzymes of thiosulfate oxidation such as rhodanese and sulfite oxidase activities were detected in cell-free extracts of M. goesingense CBMB5, and M. fujisawaense CBMB37, and thiosulfate oxidase (tetrathionate synthase) activity was not observed. It indicated that both the organisms use the "non-S4 intermediate" sulfur oxidation pathway for thiosulfate oxidation. It is concluded from this study that M. goesingense CBMB5, and M. fujisawaense CBMB37 exhibited mixotrophic metabolism in medium containing methanol plus thiosulfate and that thiosulfate oxidation and the presence of a "Paracoccus sulfur oxidation" (PSO) pathway in methylotrophic bacteria are species dependant.

Characterization of plant growth-promoting traits of bacteria isolated from larval guts of diamondback moth Plutella xylostella (lepidoptera: plutellidae).

Eight bacterial isolates from the larval guts of Diamondback moths (Plutella xylostella) were tested for their plant growth-promoting (PGP) traits and effects on early plant growth. All of the strains tested positive for nitrogen fixation and indole 3-acetic acid (IAA) and salicylic acid production but negative for hydrogen cyanide and pectinase production. In addition, five of the isolates exhibited significant levels of tricalcium phosphate and zinc oxide solubilization; six isolates were able to oxidize sulfur in growth media; and four isolates tested positive for chitinase and beta-1,3-glucanase activities. Based on their IAA production, six strains including four that were 1-aminocyclopropane-1-carboxylate (ACC) deaminase positive and two that were ACC deaminase negative were tested for PGP activity on the early growth of canola and tomato seeds under gnotobiotic conditions. Acinetobacter sp. PSGB04 significantly increased root length (41%), seedling vigor, and dry biomass (30%) of the canola test plants, whereas Pseudomonas sp. PRGB06 inhibited the mycelial growth of Botrytis cinerea, Colletotrichum coccodes, C. gleospoiroides, Rhizoctonia solani, and Sclerotia sclerotiorum under in vitro conditions. A significant increase, greater than that of the control, was also noted for growth parameters of the tomato test plants when the seeds were treated with PRGB06. Therefore, the results of the present study suggest that bacteria associated with insect larval guts possess PGP traits and positively influence plant growth. Therefore, insect gut bacteria as effective PGP agents represent an unexplored niche and may broaden the spectrum of beneficial bacteria available for crop production.

Cultivable bacteria associated with larval gut of prothiofos-resistant, prothiofos-susceptible and field-caught populations of diamondback moth, Plutella xylostella and their potential for, antagonism towards entomopathogenic fungi and host insect nutrition.

AIMS: To evaluate whether the gut bacteria of insecticide-resistant, insecticide-susceptible and field-caught populations of the lepidopteran insect pest diamondback moth (DBM)--Plutella xylostella (L.)--are variable and their role in host protection and nutrition. METHODS AND RESULTS: The gut bacterial populations of the three DBM larvae populations were found to be significantly different, irrespective of the developmental stage. The 16S rRNA gene sequence analysis of the DBM gut bacteria revealed that the bacterial population from the prothiofos-resistant larval gut was more diversified with Pseudomonas sp., Stenotrophomonas sp., Acinetobacter sp., and Serratia marcescens. Meanwhile, the susceptible larvae were associated with Brachybacterium sp., Acinetobacter sp. and S. marcescens and the field-caught population harboured a rather simple gut microflora of phylotypes belonging to Serratia. The siderophore-producing Pseudomonas sp. strain PRGB06 showed antagonistic activity towards entomopathogenic fungi, including Beaveria bassiana, Hirsutella thompsonii, Metarhizium anisopliae, Paecilomyces sp., and Paecilomyces tenuipes, while the chitinase-producing S. marcescens enhanced the larval growth and development. CONCLUSION: There was a significant variation in the gut bacteria from the three different populations of DBM. The production of antifungal siderophore compounds, like pyoverdine, may contribute to host antagonism against entomopathogens. The production of chitinase by gut bacteria appeared to contribute to host nutrition. SIGNIFICANCE AND IMPACT OF THE STUDY: The results provide the first comprehensive description of the gut microbial communities in three different populations of an important crucifer pest DBM and suggest that the bacteria associated with the insect pest could be of interest for developing a pest management strategy.

Thiosulfate oxidation and mixotrophic growth of Methylobacterium oryzae.

Thiosulfate oxidation and mixotrophic growth with succinate or methanol plus thiosulfate was examined in nutrient-limited mixotrophic condition for Methylobacterium oryzae CBMB20, which was recently characterized and reported as a novel species isolated from rice. Methylobacterium oryzae was able to utilize thiosulfate in the presence of sulfate. Thiosulfate oxidation increased the protein yield by 25% in mixotrophic medium containing 18.5 mmol.L-1 of sodium succinate and 20 mmol.L-1 of sodium thiosulfate on day 5. The respirometric study revealed that thiosulfate was the most preferable reduced inorganic sulfur source, followed by sulfur and sulfite. Thiosulfate was predominantly oxidized to sulfate and intermediate products of thiosulfate oxidation, such as tetrathionate, trithionate, polythionate, and sulfur, were not detected in spent medium. It indicated that bacterium use the non-S4 intermediate sulfur oxidation pathway for thiosulfate oxidation. Thiosulfate oxidation enzymes, such as rhodanese and sulfite oxidase activities appeared to be constitutively expressed, but activity increased during growth on thiosulfate. No thiosulfate oxidase (tetrathionate synthase) activity was detected.