Unstandardized UV-C dose used for killing harmful cyanobacteria may instead initiate accelerated growth in the target organisms.

Although UV-C radiation has been in use for killing unwanted cyanobacteria, experiments with lower doses of UV-C radiation instead showed induction of growth related parameters and enhanced biomass production in the cyanobacterium Nostoc muscorum Meg1. When the cyanobacterial cultures were exposed to UV-C radiation of varying doses (6, 12 and 18 mJ/cm2), concentrations of various photo-absorbing pigments, RuBisCO and D1 protein of PSII; activities of oxygen evolving complex, nitrogenase and glutamine synthetase were significantly increased upon 6 and 12 mJ/cm2 UV-C radiation exposures. Resulting higher photosynthetic performance was evident from the augmentation in carbohydrate content by ∼49% under single exposure to 6 mJ/cm2 UV-C by fifteenth day. The increased performances of both RuBisCO and D1 proteins were in part also due to induction at the genetic level as seen from the increase in their mRNA and protein levels under treatment. Similar increase was also observed in protein (16%) and in lipid contents (43%) that reflected an upsurge in the total biomass. Highest biomass (463 mg/L/d) was noted in culture exposed to 6 mJ/cm2 UV-C radiation, representing a ∼25% increase. Furthermore the possibility of this organism using part of the incident UV-C radiation as an additional source of energy was deduced from an experiment where the thylakoid membranes excited within UV (226-400 nm) range showed emission at longer wavelengths with an emission maximum at ∼640 nm. Thus this work provides evidence that lower UV-C doses can potentially augment cyanobacterial growth and use of unstandardized UV-C doses for restricting cyanobacterial growth may in fact produce contrary result.

Modulation of oxidant and antioxidant homeostasis in the cyanobacterium Nostoc muscorum Meg1 under UV-C radiation stress.

The present work was conducted to study how restoration of perturbed oxidant and antioxidant homeostasis is achieved in the UV-C radiation exposed cells of cyanobacterium Nostoc muscorum Meg1. Exposure to varying doses of UV-C radiation (6, 12, 18 and 24 mJ/cm2) showed damage to ultrastructures especially cytoplasmic membrane, cell wall and organisation of thylakoid membranes of the cyanobacterium under transmission electron microscope (TEM). All doses of UV-C exposure significantly induced most of the enzymatic antioxidant {catalase, superoxide dismutase (SOD) and glutathione reductase (GR)} activities, their protein levels (western blot analysis) and mRNA levels (real time PCR analysis) within the first hour of post UV-C radiation incubation period. In the same way, contents of many non-enzymatic antioxidants such as ascorbic acid, reduced glutathione, proline, phenol and flavonoids were also augmented in response to such UV-C radiation exposure. Although notable increase in ROS level was only seen in cultures treated with 24 mJ/cm2 UV-C exposure which also registered increase in protein oxidation (22%) and lipid peroxidation (20%), this boost in both enzymatic and non-enzymatic antioxidants was significant in all radiation exposed cells indicating cell's preparation to combat rise in oxidants. Further, albeit all antioxidants increased considerably, their levels were restored back to control values by day seventh re-establishing physiological redox state for normal metabolic function. The combined efficiency of the enzymatic and non-enzymatic antioxidants were so effective that they were able to bring down the increase levels of ROS, lipid peroxidation and protein oxidation to the physiological levels within 1 h of radiation exposure signifying their importance in the defensive roles in protecting the organism from oxidative toxicity induced by UV-C radiation exposure.

Dose dependent variance in UV-C radiation induced effects on carbon and nitrogen metabolism in the cyanobacterium Nostoc muscorum Meg1.

With the intention of getting an insight into the differential effect of UV-C radiation on the N2-fixing heterocystous cyanobacterium Nostoc muscorum Meg1, various aspects of carbon and nitrogen metabolism was evaluated in the organism. Exposure to different doses of UV-C (6, 12, 18 and 24 mJ/cm2) showed that among various photo-absorbing pigments, phycobiliproteins were most sensitive. Oxygen evolving complex (OEC) activity measured as net oxygen evolution rate decreased by 63% upon 24 mJ/cm2 exposure. Western blot analysis established that D1 protein of PSII was highly sensitive and its levels decreased even at a radiation dose as low as 6 mJ/cm2. In contrast, levels of the Calvin cycle enzyme RuBisCO was increased at 6 and 12 mJ/cm2 doses but the level decreased drastically (84%) at higher dose (24 mJ/cm2). The nitrogenase enzyme activity decreased at all doses but the ammonia assimilating enzyme glutamine synthetase (GS) activity recorded increase at the lower doses. The reactive oxygen species (ROS) and lipid peroxidation increased upon UV-C exposure. Transmission electron microscopic observation revealed damage to ultrastructure especially the thylakoid membrane organization, aggregation of dissolving phycobilisomes and loss of caboxysomes. Interestingly, sub-lethal radiation (6 and 12 mJ/cm2) dose exposures increased the growth rate in the organism when growth was measured over a period of 11 days after radiation exposure.

Diazotrophic specific cytochrome c oxidase required to overcome light stress in the cyanobacterium Nostoc muscorum.

Diazotrophic, filamentous and heterocystous cyanobacterium Nostoc muscorum perform photosynthesis in vegetative whereas nitrogen fixation occurs in heterocyst only. However, despite their metabolic plasticity, respiration takes place both in vegetative cells and heterocysts. The role of the respiratory electron transport system and terminal oxidases under light stress is not evident so far. As compared to the diazotrophically grown cultures, the non-diazotrophically grown cultures of the N. muscorum show a slight decrease in their growth, chlorophyll a contents and photosynthetic O2 evolution under light stress. Whereas respiratory O2 uptake under identical stress condition increases several fold. Likewise, nitrogen fixing enzyme i.e. nitrogenase over-expresses itself under light stress condition. The terminal enzyme of respiratory electron transport chain i.e. cytochrome c oxidase shows more activity under light stress, whilst light stress has no impact on Ca(++)-dependent ATPase activity. This leads to the conclusion that under light stress, cytochrome c oxidase plays a vital role in mitigating given light stress.

Differential display of antioxidants in mitigating adverse effects of UV-B radiation in Nostoc muscorum and Phormidium foveolarum photoacclimated to different irradiances.

The response of UV-B radiation (UV-BL; 0.5) and (UV-BH; 1.5 µmol m(-2) s(-1)) was investigated in two cyanobacteria--Nostoc muscorum and Phormidium foveolarum--preacclimated to low, normal, and high (LL; 15 ± 0.5, NL; 75 ± 3.5, and HL; 225 ± 5.5 µmol photon m(-2) s(-1), respectively) light intensities. Acclimation to HL stimulated growth over the control; however, both UV-B doses induced decline in growth of both cyanobacteria. UV-B-induced decline was maximum in LL acclimated cells which was in consonance with the results for chlorophyll content. LL acclimation led to an increase in phycocyanin content. Carotenoids content was maximum in HL acclimated (UV-B untreated and treated) cells of N. muscorum and P. foveolarum. Maximum stimulation in SOR and H2O2 levels was observed under LL + UV-BH treatment which was correlated with inefficient antioxidative mechanism (SOD, CAT, POD, and GST) in these cells. MDA and RCG contents also showed a similar trend. Mitigation of UV-B-induced stress response could be established in HL acclimated cells due to the concerted and differential display of enzymatic and nonenzymatic antioxidants in them. P. foveolarum proved to be more resistant than N. muscorum under the studied test conditions.

Low and high doses of UV-B differentially modulate chlorpyrifos-induced alterations in nitrogen metabolism of cyanobacteria.

The present study assessed the comparative responses on the specific growth rate, nitrogen metabolism and enzymes associated with nitrogen metabolism in two nitrogen fixing cyanobacteria-Nostoc muscorum and Phormidium foveolarum exposed to two UV-B doses (low; UV-BL: 0.5472kJm(-2) and high; UV-BH: 5.472kJm(-2)) and two doses of the insecticide chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate; low i.e. CPL, 1µgml(-1) and high i.e. CPH, 2µgml(-1)) singly and in combination. The specific growth rate, NO3(-) and NO2(-) uptake, nitrate assimilating enzymes - nitrate reductase and nitrite reductase and ammonium assimilating enzymes - glutamine synthetase and glutamate synthase were severely affected when treated either with CPH or/and UV-BH while glutamate dehydrogenase exhibited a stimulatory response. CPL also reduced all the measured parameters (except GDH activity) after 24h, however, a stimulatory effect was observed after 72h due to an increase in nitrogen metabolism (and other antioxidant) enzymes during this period. UV-BL did not cause significant alteration in the studied parameters while in combination with CP doses, it either alleviated the inhibitory effects or further enhanced the CPL induced activities of these enzymes (except GDH). Overall results indicate the resistant nature of P. foveolarum against the inhibitory doses of UV-B and chlorpyrifos in comparison to N. muscorum.

Compatibility of ascorbate-glutathione cycle enzymes in cyanobacteria against low and high UV-B exposures, simultaneously exposed to low and high doses of chlorpyrifos.

This study deals with the comparative responses of the two cyanobacteria viz. Nostoc muscorum and Phormidium foveolarum against single and combined doses of low (UV-B(L,) 0.1 µmol m(-2) s(-1)) and high (UV-B(H), 1.0 µmol m(-2) s(-1)) fluence rates of ultraviolet-B radiation with low (CP(L), 1 µg ml(-1)) and high (CP(H), 2 µg ml(-1)) doses of the insecticide chlorpyrifos by measuring changes in growth, ascorbate-glutathione cycle enzymes and related metabolites. CP(L) and UV-B(L) both caused lesser increase in ROS but significantly stimulated AsA-GSH cycle enzymes. On the other hand, CP(H) and UV-B(H) posed inhibitory effects by enhancing ROS and inhibiting AsA-GSH cycle enzymes. Inhibitions in CP(H) or UV-B(H) treated samples were significantly prevented when they were supplemented with UV-B(L) and CP(L) (after 72 h), respectively by lowering down ROS and enhancing AsA-GSH enzymes and related metabolites which manifested in terms of improved biomass accumulation.

High light intensity augments mercury toxicity in cyanobacterium Nostoc muscorum.

The present study is aimed at investigating the role of growth irradiance in determining the extent of mercury (Hg) toxicity on various physiological parameters viz. growth, pigment contents, photosynthesis, respiration, (14)CO(2) fixation, photosynthetic electron transport, photorespiration and enzyme activity of cyanobacterium Nostoc muscorum. A general decline was observed in all these parameters with increasing concentration of Hg except for carotenoids content and respiratory activity which exhibited significant enhancement. This effect was more pronounced in high light (130 µmol photon m(-2) s(-1)) exposed cells as compared to normal (70 µmol photon m(-2) s(-1)) and low (10 µmol photon m(-2) s(-1)) light exposed cells. Among the photosynthetic electron transport activities, whole chain was found to be more sensitive than photosystem II (PSII) and photosystem I (PSI). (14)CO(2) fixation was more affected as compared to O(2) evolution when exposed to Hg and different light intensities. Photorespiratory activity, which is an index of protecting organisms from light-induced damage, also showed a similar declining trend. Enzyme assay revealed that among the carboxylating enzymes, activity of RUBISCO was more severely inhibited than PEPCase. Thus, these results suggest that Hg itself was toxic at all tested concentrations and high light intensity augmented its toxicity in N. muscorum inhibiting the growth, pigment contents and photosynthetic activity of the organism.

UV-B induced differential effect on growth and nitrogen metabolism in two cyanobacteria under copper toxicity.

In the present study, impact of low (UV-B(L): 0.1 µmol m(-2) s(-1)) and high (UV-BH: 1.0 µmol m(-2) s(-1)) fluence rates of ultraviolet-B on growth and nitrogen metabolism in two cyanobacteria: Phormidium foveolarum and Nostoc muscorum under copper toxicity (2 and 5 µM) was investigated after 24 and 72 h of experiments. Copper and UV-BH treatment suppressed growth but more in N. muscorum which was accompanied by significant accumulation of Cu. Nitrate and nitrite uptake rates and activities of nitrogen assimilating enzymes i.e. nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS) and glutamate synthase (GOGAT) except glutamate dehydrogenase activity (GDH; aminating) were decreased following treatments of Cu and UV-BH, and under combined treatments the effect was greater. On contrary, UV-BL declined Cu toxicity significantly. The study concludes that Cu and UV-BH suppressed the activity of NR, NiR, GS and GOGAT (except GDH) hence decreased growth. However, UV-BL showed cross tolerance in test organisms against Cu toxicity up to certain extent. Phormidium foveolarum is comparatively less sensitive against UV-BH and excess Cu, a situation likely exists in nature, hence it may be used as a biofertilizer for sustainable agriculture.

Differential physiological and biochemical responses of two cyanobacteria Nostoc muscorum and Phormidium foveolarum against oxyfluorfen and UV-B radiation.

In the present study, degree of tolerance and tolerance strategies of two paddy field cyanobacteria viz. Nostoc muscorum and Phormidium foveolarum against oxyfluorfen (10 and 20 µg ml(-1)) and UV-B (7.2 kJ m(-2)d(-1)) stress were investigated. Oxyfluorfen and UV-B decreased growth, photosynthesis, nutrient uptake, nitrate reductase, acid and alkaline phosphatase activities, which accompanied with the increase in the level of oxidative stress. However, growth was more affected in N. muscorum than P. foveolarum. Antioxidants exhibited differential responses against oxyfluorfen and UV-B stress. Ascorbate and proline levels were higher in P. foveolarum. A protein of 66 kDa was expressed in N. muscorum, however, it was absent in P. foveolarum than those of N. muscorum. Besides this, a protein of 29 kDa appeared in P. foveolarum under all the treatments, but it was present only in control cells of N. muscorum cells. Overall results indicated resistant nature of P. foveolarum against oxyfluorfen and UV-B stress in comparison to N. muscorum.