Comparison between Conventional Bacterial and Algal-Bacterial Aerobic Granular Sludge Systems in the Treatment of Saline Wastewater

The increasing generation of saline wastewater through various industrial activities is becoming a global concern for activated sludge (AS) based biological treatment which is widely applied in wastewater treatment plants (WWTPs). As for the AS process, an increase in wastewater salinity has negative impact on its overall performance. The advent of conventional aerobic granular sludge (AGS) or bacterial AGS biotechnology has gained much attention because of its superior performance. The development of algal-bacterial AGS could enhance better nutrients removal, potentially reduce aeration cost through symbiotic algae-bacterial activity, and thus, can also reduce overall treatment cost. Nonetheless, the potential of salt stress to decrease biomass growth, microbial activity and nutrient removal exist. Up to the present, little information is available on saline wastewater treatment by algal-bacterial AGS. To the authors’ best knowledge, a comparison of the two AGS systems has not been done to evaluate nutrients removal capacity in the context of salinity increase. This study sought to figure out the impact of salinity on the algal-bacterial AGS system in comparison to bacterial AGS one, contributing to the application of AGS technology in the real world of saline wastewater treatment. In this study, the salt concentrations tested were 0 g/L, 1 g/L, 5 g/L, 10 g/L and 15 g/L of NaCl with 24-hr artificial illuminance of approximately 97.2 µmol m¯²s¯¹, and mature bacterial and algal-bacterial AGS were used for the operation of two identical sequencing batch reactors (SBRs) with a working volume of 0.9 L each, respectively. The results showed that salinity increase caused no apparent change in the color of bacterial AGS; while for algal-bacterial AGS, its color was progressively changed from green to dark green. A consequent increase in granule diameter and fluffiness was observed in the bacterial AGS reactor with the increase of salinity in comparison to a decrease in algal-bacterial AGS diameter. However, nitrite accumulation peaked from 1.0 mg/L and 0.4 mg/L at 1 g/L NaCl in the bacterial and algal-bacterial AGS systems, respectively to 9.8 mg/L in both systems when NaCl concentration varied from 5 g/L to 15 g/L. Almost no ammonia nitrogen was detected in the effluent except at 10 g/L NaCl concentration, where it averaged 4.2 mg/L and 2.4 mg/L, respectively, in the bacterial and algal-bacterial AGS systems. Nutrients removal in the algal-bacterial system was relatively higher than the bacterial AGS in terms of nitrogen and phosphorus removals. Nonetheless, the nutrient removal rate was almost 50% or lower. Results show that algal-bacterial AGS is more adaptable to salinity increase and could be more suitable for saline wastewater treatment. Optimization of operation conditions for algal-bacterial AGS system would be important to ensure its stably high efficiency in practice.

Adaptive Responses of Carum copticum to in vitro Salt Stress

Salinity is one of the most widespread agricultural problems in arid and semi-arid areas that limits the plant growth and crop productivity. In this study, the salt stress effects on protein, reducing sugar, proline contents and antioxidant enzymes activities of Carum copticum L. under in vitro conditions were studied. Seeds of C. copticum were cultured in Murashige and Skoog (MS) medium containing 0, 25, 50, 100 and 150 mM NaCl and calli were cultured in MS medium containing 1 μM 2, 4-dichlorophenoxyacetic acid, 4 μM benzyl amino purine and different levels of NaCl (0, 25, 50, 100 and 150 mM). After NaCl treatment for 28 days, the proline and reducing sugar contents of shoots, roots and calli increased significantly in relation to the severity of the salt stress. The highest amount of proline and carbohydrate were observed at 150 and 100 mM NaCl, respectively. The reducing sugar accumulation in shoots was the highest as compared to roots, whereas, proline contents did not show any significant difference in roots and shoots under salt stress. The results showed significant reduction of protein contents in seedlings and calli. Based on these results, proteins extracted from the shoots, roots and calli of C. copticum treated with 150 mM NaCl showed the lowest contents. The positive relationships were observed between activity of antioxidant enzymes and the increase in stress levels. Catalase, ascorbate peroxidase and superoxide dismutase activity increased significantly under salt concentrations in comparison to the control. These results suggest that the accumulation of proline and sugars, and activation of antioxidant enzymes play adaptive roles in the adaptation of seedlings and callus of C. copticum to saline conditions.

Effect of Two Different Biochars on Germination and Seedlings Growth of Salad, Cress and Barley

The application of biochar to soils is becoming more and more common. Its application which is generally reported to improve the physical, chemical, and biological properties of soils, has an indirect effect on soil health and increased crop yields. However, many of the previous results are highly variable and dependent mainly on the initial soil properties, biochar characteristics, and production conditions. In this study, two biochars which are biochar II (BC II) derived from a blend of paper sludge and wheat husks and biochar 005 (BC 005) derived from sewage sludge with a KCl additive, are used, and the physical and chemical properties of BC II are characterized. To determine the potential impact of salt stress and toxic and volatile substances, the second part of this study focused on the effect biochars have on germination of salad (Lactuca sativa L.), barley (Hordeum vulgare), and cress (Lepidium sativum) respectively. Our results indicate that Biochar II showed some unique properties compared to the soil, such as high EC, high content of K, Na, Mg, and low content of heavy metals. Concerning salad and barley germination test, no negative effect of BC II and BC 005 was observed. However, a negative effect of BC 005 at 8% level was revealed. The test of the effect of volatile substances on germination of cress revealed a positive effect of BC II, while a negative effect was observed for BC 005. Moreover, the water holding capacities of biochar-sand mixtures increased with increasing biochar application. Collectively, BC II could be safely used for agriculture and could provide the potential for a better plant growth.

Alleviation of Adverse Effects of Salt Stress on Soybean (Glycine max. L.) by Using Osmoprotectants and Organic Nutrients

Salinity is one of the major factors limiting crop production in an arid environment. Despite its global importance soybean production suffer the problems of salinity stress causing damages at plant development. So it is implacable to either search for salinity enhancement of soybean plants. Therefore, in the current study we try to clarify the mechanism that might be involved in the ameliorating effects of osmo-protectants such as proline and glycine betaine as well as, compost application on soybean plants grown under salinity stress. The experiment was conducted under greenhouse conditions at the Graduate School of Biosphere Science Laboratory of Hiroshima University, Japan in 2011. The experiment was designed as a spilt-split plot based on randomized complete block design with four replications. The treatments could be summarized as follows; (i) salinity concentrations (0 and 15 mM), (ii) compost treatments (0 and 24 t ha-1) and (iii) the exogenous, proline and glycine betaine concentrations (0 mM and 25 mM) for each. Results indicated that salinity stress induced reduction in growth and physiological aspects (dry weight per plant, chlorophyll content, N and K+ content) of soybean plant compared with those of the unstressed plants. On the other hand, salinity stress led to increases in the electrolyte leakage ratio, Na and proline contents. Special attention was paid to, the tolerance against salt stress was observed, the improvement of salt tolerance resulted from proline, glycine betaine and compost were accompanied with improved K+, and proline accumulation. While, significantly decreased electrolyte leakage ratio and Na+ content. These results clearly demonstrate that harmful effect of salinity could reduce on growth aspects of soybean. Consequently, exogenous osmoprotectants combine with compost will effectively solve seasonal salinity stress problem and are a good strategy to increase salinity resistance of soybean in the drylands.

A Novel Gene Encoding Ankyrin-Repeat Protein, SHG1, is Indispensable for Seed Germination under Moderate Salt Stress

Salt stress adversely affects plant growth at various stages of development including seed germination, seedling establishment, vegetative growth and finally reproduction. Because of their immobile nature, plants have evolved mechanisms to sense and respond to salt stress. Seed dormancy is an adaptive trait that enables seed germination to coincide with favorable environmental conditions. We identified a novel locus of Arabidopsis, designated SHG1 (salt hypersensitive germination 1), whose disruption leads to reduced germination rate under moderate salt stress conditions. SHG1 encodes a transmembrane protein with an ankyrin-repeat motif that has been implicated in diverse cellular processes such as signal transduction. The shg1-disrupted Arabidopsis mutant died at the cotyledon stage when sown on salt-containing medium, although wild-type plants could form true leaves under the same conditions. On the other hand, this mutant showed similar phenotypes to wild-type plants when sown on medium without salt and transferred to salt-containing medium at the vegetative stage. These results suggested that SHG1 played indispensable role in the seed germination and seedling establishment under moderate salt stress conditions. SHG1 may be involved in the release of seed dormancy.

Seed Treatment during Germination in Linseed to Overcome Salt and Drought Stresses (Linum usitatissimum L.)

Evaluation of crop plants resistance to environmental stresses specially in germination stage is a critical factor in their selection in different conditions of cultivation. Therefore use of a procedure in controllable situation can help to evaluate plants reaction to stress quickly and precisely. In order to study germination characteristics of flax in water and salinity stress conditions were conducted two laboratories experimental. The two experimental were conducted in 4-replicant completing random design for salinity and water stress. The treatment, for salinity and water stress was three potential (zero, 40, 80 mM) of NaCl and three potential (zero, -2, -4 bar) of PEG respectively. Germination percentage and rate, in addition to Radical and plumule length and dry-weight and plumule/Radical ration were measured. All of characteristics reduce under water stress conditions. salinity stress significant reduce germination rate and Radical and plumule length of flax seeds. Hydropriming and osmopriming significant increased germination rate, plumule length and plumule/Radical ration ration of flax seeds. But germination percentage and Radical and plumule dry weight significant increased only in hydropriming treat. Hydropriming and osmopriming could not be used to improved germination under saline and drought stress. But has more tolerance in salinity and drought stress in flax by less reduce in Radical and plumule length under saline and drought stress.

Growth and Stomatal Responses of Bread Wheat Genotypes in Tolerance to Salt Stress

Plant growth is affected by the osmotic stress as well as toxicity of salt in leaves. In order to study of salt stress effects on stomatal conductance and growth rate and relationship between them as wells osmotic and Na+-specific effects on these traits, four bread wheat genotypes differing in salt tolerance were selected. Salinity was applied when the leaf 4 was fully expanded. Sodium (Na+) concentrations in flag leaf blade at 3 salinity levels (0, 100 and 200 mM NaCl) were measured. Salt-tolerant genotypes showed higher stomatal conductance and growth rate compared to salt-sensitive ones. After 10 and 20 days exposure to salt, stomatal conductance and relative growth rate were reduced, but the reduction was greater in sensitive genotypes. Growth rate was reduced severely in the first period (1-10 days) of salt commencements and it was due to osmotic effect of salt not Na+ toxicity. In the second period (11-20 days) after salt treatment growth reduced only when salt accumulated to toxic concentrations in the leaves. A positive relationship between stomatal conductance and relative growth rate showed that stomatal conductance can be a reliable indicator of growth rate, and finally can be considered as a sensitive indicator of the osmotic stress. It seems 20 days after salinity, the major effect of salt, especially at low to moderate salinity levels on growth properties was due to the osmotic effect of salt, not to Na+-specific effects within the plant.

Rice cDNA Encoding PROLM is Capable of Rescuing Salt Sensitive Yeast Phenotypes G19 and Axt3K from Salt Stress

Rice seed expression (cDNA) library in the Lambda Zap 11® phage constructed from the developing grain 10-20 days after flowering was transformed into yeast for functional complementation assays in three salt sensitive yeast mutants S. cerevisiae strain CY162, G19 and Axt3K. Transformed cells of G19 and Axt3K with pYES vector with cDNA inserts showed enhance tolerance than those with empty pYes vector. Sequencing of the cDNA inserts revealed that they encode for the putative proteins with the sequence homologous to rice putative protein PROLM24 (Os06g31070), a prolamin precursor. Expression of this cDNA did not affect yeast growth in absence of salt. Axt3k and G19 strains expressing the PROLM24 were able to grow upto 400 mM and 600 mM of NaCl respectively. Similarly, Axt3k mutant with PROLM24 expression showed comparatively higher growth rate in the medium with excess LiCl (50 mM). The observation that expression of PROLM24 rescued the salt sensitive phenotypes of G19 and Axt3k indicates the existence of a regulatory system that ameliorates the effect of salt stress in the transformed yeast mutants. However, the exact function of the cDNA sequence, which shows partial sequence homology to yeast UTR1 is not clear. Although UTR1 involved in ferrous uptake and iron homeostasis in yeast cells, there is no evidence to prove its role in Na+ homeostasis in yeast cells. Absence of transmembrane regions in Os06g31070 protein indicates that salt tolerance is achieved not through the direct functional complementation of the mutant genes but through an alternative mechanism.

Effects of Wastewater Strength and Salt Stress on Microalgal Biomass Production and Lipid Accumulation

This work aims to investigate a potential of microalgae for utilizing industrial wastewater as a cheap nutrient for their growth and oil accumulation. Wastewater was collected from the effluent ponds of agro-industrial factories (cassava and ethanol production plants). Only 2 microalgal strains were isolated and identified as Scenedesmus quadricauda and Chlorella sp.. However, only S. quadricauda was selected to cultivate in various wastewater concentrations (10%, 20%, 40%, 60%, 80% and 100%). The highest biomass obtained at 6.6×106 and 6.27×106 cells/ml when 60% wastewater was used in flask and photo-bioreactor. The cultures gave the highest lipid content at 18.58 % and 42.86% in cases of S. quadricauda and S. obliquus. In addition, under salt stress (1.0 M NaCl), S. obliquus demonstrated the highest lipid content at 50% which was much more than the case of no NaCl adding. However, the concentration of NaCl does not affect on lipid accumulation in case of S. quadricauda.

Hydropriming and Osmopriming Effects on Cumin(Cuminum Cyminum L.) Seeds Germination

In production of medicinal plants, seed germination is very important problem. The treated seeds (control, hydro priming and ZnSO4) of Cumin (Cuminum cyminum L.) were evaluated at germination and seedling growth for tolerance to salt (NaCl and Na2SO4) conditions at the same water potentials of 0.0, -0.3, -0.6, - 0.9 and -1.2MPa. Electrical conductivity (EC) values of the NaCl solutions were 0.0, 6.5, 12.7, 18.4 and 23.5 dSm-1, respectively. The objective of the study was to determine factors responsible for germination and early seedling growth due to salt toxicity or osmotic effect and to optimize the best priming treatment for these stress conditions. Results revealed that germination delayed in both solutions, having variable germination with different priming treatments. Germination, shoot and weight, root and shoot length were higher but mean germination time and abnormal germination percentage were lower in NaCl than Na2SO4 at the same water potential. The root / shoot weight and R/S length increased with increase in osmotic potential in both NaCl and Na2SO4 solutions. NaCl had less inhibitor effect on seedling growth than the germination. It was concluded that inhibition of germination at the same water potential of NaCl and Na2SO4 resulted from salt toxicity rather than osmotic effect. Hydro priming increased germination and seedling growth under salt stress. This protocol has practical importance and could be recommended to farmers to achieve higher germination and uniform emergence under field conditions.

Chlorophyll Fluorescence as Criterion for the Diagnosis Salt Stress in Wheat (Triticum aestivum) Plants

To investigate effect of salt stress on Chlorophyll fluorescence four cultivars (fong,star,chamran and kharchia) of wheat (Triticum aestivum) plants subjected to salinity levels ( control,8,12 and 16 dsm-1 ) from one week after emergence to the end of stem elongation under greenhouse condition . results showed that quantum yield of photosystem II from light adopted leaves (ΦPSII), Photochemical quenching (qP) ,quantum yield of dark adopted leaves (fv/fm) and non photochemical quenching (NPq) were affected by salt stress . Salinity levels affected photosynthetic rate. Star and fong cultivars showed minimum and maximum levels of photosynthetic rate in respectively. Minimum photosynthetic rate differences between levels of salinity were shown in Kharchia. Shoot dry matter of all cultivars decreased by increasing salinity levels. Results showed that non photochemical quenching by salinity levels attribute to the decreases in shoot dry matter.