An insight into algicidal characteristics of Bacillus altitudinis G3 from dysfunctional photosystem and overproduction of reactive oxygen species.

Cyanobacterial blooms negatively affect aquatic ecosystems and human health. Algicidal bacteria can efficiently kill bloom-causing cyanobacteria. Bacillus altitudinis G3 isolated from Dianchi Lake shows high algicidal activity against Microcystis aeruginosa. In this study, we investigated its algicidal characteristics including attack mode, photosynthesis responses, and source and the contribution of reactive oxygen species (ROS). The results showed that G3 efficiently and specifically killed M. aeruginosa mainly by releasing both thermolabile and thermostable algicidal substances, which exhibited the highest algicidal activity (99.8%, 72 h) in bacterial mid-logarithmic growth phase. The algicidal ratio under full-light conditions (99.5%, 60 h) was significantly higher than under dark conditions (<20%, P < 0.001). G3 filtrate caused photosystem dysfunction by decreasing photosynthetic efficiency, as indicated by significantly decreased Fv/Fm and PIABS (P < 0.001) values. It also inhibited photosynthetic electron transfer as indicated by significantly decreased rETR (P < 0.001), especially QA- downstream, as revealed by significantly decreased φEo and ψo, and increased Mo (P < 0.001). These results indicated that the algicidal activity of G3 filtrate is light-dependent, and the cyanobacterial photosystem is an important target. Cyanobacterial ROS and malondialdehyde contents greatly increased by 37.1% and 208% at 36 h, respectively. ROS levels decreased by 49.2% (9 h) when diuron (3-(3-4-dichlorophenyl)-1,1-dimethylurea) partially blocked photosynthetic electron transport from QA to QB. Therefore, excessive ROS were produced from disrupted photosynthesis, especially the inhibited electron transport area in QA- downstream, and caused severe lipid peroxidation with significantly increased MDA content and oxidative stress in cyanobacteria. The ROS scavenger N-acetyl-l-cysteine significantly decreased both cyanobacterial ROS levels (34%) and algicidal ratio (52%, P < 0.05) at 39 h. Thus, excessive ROS production due to G3 filtrate administration significantly contributed to its algicidal effect. G3 could be an excellent algicide to control M. aeruginosa blooms in waters under suitable light conditions.

Algicidal effect of tryptoline against Microcystis aeruginosa: Excess reactive oxygen species production mediated by photosynthesis.

Cyanobacterial blooms significantly decrease water quality and can damage ecosystems and, as such, require efficient control methods. Algicidal bacteria and their associated substances are promising tools for controlling cyanobacterial blooms; however, their specific algicidal mechanisms remain unclear. Therefore, the current study sought to investigate the algicidal mechanism of tryptoline (1,2,3,4-tetrahydro-9 h-pyrido[3,4-b]indole) against Microcystis aeruginosa, with a specific focus on the contribution made by reactive oxygen species (ROS), the underlying mechanisms of ROS increase, as well as the photosystem response. Results show that the algicidal ratio of tryptoline significantly and positively correlates with algal ROS. Moreover, 93.79% of the algicidal ratio variation is attributed to ROS in the tryptoline group, while only 47.75% can be attributed to ROS in the tryptoline + N-acetyl-L-cysteine (NAC) group, where ROS are partially scavenged by NAC. In the presence of tryptoline, algicidal effect and ROS levels were significantly enhanced in the presence of light as compared to those in the dark (P < 0.001). Hence, the increase in ROS production attributed to tryptoline is primarily affected by the presence of light and photosynthesis. Additionally, tryptoline significantly reduces Fv/Fm, PIABS, ETo/RC, and the expression of psaB and psbA genes related to photosynthesis, while increasing Vj and DIo/RC (P < 0.05). These results suggest that tryptoline hinders algal photosynthesis by significantly decreasing photosynthetic efficiency and carbon assimilation, inhibiting photochemical electron transfer, and increasing closed reaction centers and energy loss. Moreover, following partial blockade of the photosynthetic electron transfer from QA to QB by diuron (3-(3-4-dichlorophenyl)-1,1-dimethylurea), the ROS of algae exposed to tryptoline is significantly decreased. Thus, tryptoline inhibits electron transfer downstream of QA, which increase the number of escaping electron and thereby increase ROS generation. Collectively, this study describes the algicidal mechanism of tryptoline against M. aeruginosa and highlights the critical factors associated with induction of algicidal activity.

The potential assessment of green alga Chlamydomonas reinhardtii CC-503 in the biodegradation of benz(a)anthracene and the related mechanism analysis.

Benz(a)anthracene (BaA) is a polycyclic aromatic hydrocarbons (PAHs), that belongs to a group of carcinogenic and mutagenic persistent organic pollutants found in a variety of ecological habitats. In this study, the efficient biodegradation of BaA by a green alga Chlamydomonas reinhardtii (C. reinhardtii) CC-503 was investigated. The results showed that the growth of C. reinhardtii was hardly affected with an initial concentration of 10 mg/L, but was inhibited significantly under higher concentrations of BaA (>30 mg/L) (p < 0.05). We demonstrated that the relatively high concentration of 10 mg/L BaA was degraded completely in 11 days, which indicated that C. reinhardtii had an efficient degradation system. During the degradation, the intermediate metabolites were determined to be isomeric phenanthrene or anthracene, 2,6-diisopropylnaphthalene, 1,3-diisopropylnaphthalene, 1,7-diisopropylnaphthalene, and cyclohexanol. The enzymes involved in the degradation included the homogentisate 1,2-dioxygenase (HGD), the carboxymethylenebutenolidase, the ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and the ubiquinol oxidase. The respective genes encoding these proteins were significantly up-regulated ranging from 3.17 fold to 13.03 fold and the activity of enzymes, such as HGD and Rubisco, was significantly induced up to 4.53 and 1.46 fold (p < 0.05), during the BaA metabolism. This efficient degradation ability suggests that the green alga C. reinhardtii CC-503 may be a sustainable candidate for PAHs remediation.

[Construction of multifunctional genetically engineered pesticides-degrading bacteria by homologous recombination].

Construction of multifunctional pesticides-degrading genetically engineered microorganisms (GEMs) is increasing important in the bioremediation of various pesticides contaminants in environment. However, construction of genetically stable GEMs without any exogenous antibiotic resistance is thought to be one of the bottlenecks in GEMs construction. In this article, homologous recombination vectors with the recipient's 16S rDNA as homologous recombination directing sequence (HRDS) and sacB gene as double crossover recombinants positive selective marker were firstly constructed. The methyl parathion hydroalse gene (mpd) was inserted into the 16S rDNA site of the carbofuran degrading strain Sphingomonas sp. CDS-1 by homologous recombination single crossover in the level of about 3.7 x 10-(7) - 6.8 x 10(-7). Multifunctional pesticides-degrading GEMs with one or two mpd genes inserted into the chromosome without any antibiotic marker were successfully constructed. The homologous recombination events were confirmed by PCR and southern blot methods. The obtained GEMs were genetically stable and could degrade methyl parathion and carbofuran simultaneously. The insertion of mpd gene into rrn site did not have any significant effect on recipient' s physiological and original degrading characteristics. The methyl parathion hydrolase (MPH) was expressed at a relatively high level in the recombinants and the recombinant MPH specific activity in cell lysate was higher than that of original bacterium (DLL-1) in every growth phase tested. The highest recombinant MPH specific activity was 6.22 mu/tg. In this article, we describe a first attempt to use rRNA-encoding regions of Sphingomonas strains as target site for expression of exogenous MPH, and constructed multifunctional pesticides degrading GEMs, which are genetically stable and promising for developing bioremediation strategies for the decontamination of pesticides polluted soils.