Overexpression of the Small Ubiquitin-Like Modifier protease OTS1 gene enhances drought tolerance in sugarcane (Saccharum spp. hybrid).

Sugarcane is an economically important crop plant across the globe as it is the primary source of sugar and biofuel. Its growth and development are greatly influenced by water availability; therefore, in periods of water scarcity, yields are severely compromised. Small Ubiquitin-Like Modifier (SUMO) proteases play an important role in stress responses by regulating the SUMO-related post-translational modification of proteins. In an attempt to enhance drought tolerance in sugarcane, this crop was genetically transformed with a cysteine protease (OVERLY TOLERANT TO SALT-1; OTS1) from Arabidopsis thaliana using particle bombardment. Transgenic plants were analysed in terms of photosynthetic capacity, oxidative damage, antioxidant accumulation and the SUMO-enrich protein profile was assessed. Sugarcane transformed with the AtOTS1 gene displayed enhanced drought tolerance and delayed leaf senescence under water deficit compared to the untransformed wild type (WT). The AtOTS1 transgenic plants maintained a high relative moisture content and higher photosynthesis rate when compared to the WT. In addition, when the transgene was expressed at high levels, the transformed plants were able to maintain higher stomatal conductance and chlorophyl content under moderate stress compared to the WT. Under severe water deficit stress, the transgenic plants accumulated less malondialdehyde and maintained membrane integrity. SUMOylation of total protein and protease activity was lower in the AtOTS1 transformed plants compared to the WT, with several SUMO-enriched proteins exclusively expressed in the transgenics when exposed to water deficit stress. SUMOylation of proteins likely influenced various mechanisms contributing to enhanced drought tolerance in sugarcane.

Determining biota succession in a domestic wastewater pond system after treatment with a specific consortium microalgae.

Wastewater stabilization ponds (WSPs) rely on the metabolic activities of the inhabiting microbiota to treat wastewater. A selected consortium of Chlorella vulgaris and Chlorella protothecoides were used to manipulate the natural resident microalgae assemblage to improve the treatment performance of a domestic wastewater pond treatment system in a coastal region. Since information is lacking about the resulting influence on the composition or succession of the phytoplankton or associated microbiota assemblage, the current study aimed to determine how dosing with the microalgae C. vulgaris and C. protothecoides change the efficiency of wastewater effluent treatment, as well as the composition and succession of the natural occurring phytoplankton and microbial assemblage throughout WSP system. After a year of specific microalgae inoculations, the effluent in part complied with the standards set by the Department of Water Affairs and Forestry (DWAF) and the USA, Environmental Protection Agency (EPA). The cyanobacteria Microcystis aeruginosa dominated the sixth (75%) and seventh pond (97%) before the inoculation with C. vulgaris and C. protothecoide commenced. After 12 inoculation events C. vulgaris and C. protothecoides were dominant in ponds three to seven while the dominant microbial groups were Bacteroidetes, Cyanobacteria, Firmicutes, Planctomycetes, Proteobacteria, Spirochaetes, Synergistetes and Verrucomicrobia. After the microalgae treatment, the WSP effluent were more compliant regarding to the set guidelines for effluent than prior to microalgae treatment. Based on the ability of the C. vulgaris and C. protothecoides to improve the effluent water quality, it was evident that the consortium of microalgae can be use improve domestic wastewater effluent in rural nutrient sensitive catchments. Supplementary Information: The online version contains supplementary material available at 10.1007/s40201-022-00840-z.

Treated acid mine drainage and stream recovery: Downstream impacts on benthic macroinvertebrate communities in relation to multispecies toxicity bioassays.

The success and long term effectiveness of extensive and expensive engineering solutions to restore streams impacted by Acid Mine Drainage (AMD) is rarely tested. Concentrations of pollutants were measured in water along a longitudinal gradient from a stretch of the Tweelopie stream, South Africa, that receives pH-treated acid mine drainage (AMD) from an abandoned gold mine. The biotoxic effects of treated AMD were determined through macroinvertebrate biotic indices (SASS5) and a battery of toxicity bioassays. These included the L. sativa, A. cepa, D. magna toxicity and Ames mutagenicity tests, as well as an in vitro human liver cancer cell line HepG2. Even though the Tweelopie stream was moderately to severely degraded by multiple anthropogenic stressors, the impact of the treated AMD was masked by the improvement in the system downstream after mixing with the domestic wastewater effluent receiving stream, and subsequent further dilution as a result of the karst springs downstream. The general improvement of the system downstream was clearly shown by the decrease in the ecotoxicity and mutagenicity in relation to the in-stream macroinvertebrates. PCA multivariate analysis successfully displayed associations between the different environmental variables and the decrease in toxicity and subsequent ecosystem improvement downstream. This study indicated that environmental management of AMD remediation should consider long term assessment strategies, including multiple factors, to promote biological ecosystem recovery.

River catchment responses to anthropogenic acidification in relationship with sewage effluent: An ecotoxicology screening application.

Rising environmental pressures on water resources and resource quality associated with urbanisation, industrialisation, mining and agriculture are a global concern. In the current study the upper Olifants River catchment as case study was used, to show that acid mine drainage (AMD) and acid precipitation were the two most important drivers of possible acidification during a four-year study period. Over the study period 59% of the precipitation sampled was classified as acidic with a pH value below 5.6. Traces of acidification in the river system using aquatic organisms at different trophic levels were only evident in areas of AMD point sources. Data gathered from the ecotoxicology screening tools, revealed that discharge of untreated and partially treated domestic sewage from municipal sewage treatment works and informal housing partially mitigate any traces of acidification by AMD and acid precipitation in the main stem of the upper Olifants River. The outcome of the study using phytoplankton and macroinvertebrates as indicator organisms revealed that the high loads of sewage effluent might have played a major role in the neutralization of acidic surface water conditions caused by AMD and acid precipitation. Although previous multi-stage and microcosm studies confirmed the decrease in acidity and metals concentrations by municipal wastewater, the current study is the first to provide supportive evidence of this co-attenuation on catchment scale. These findings are important for integrated water resource management on catchment level, especially in river systems with a complex mixture of pollutants.

Profiling Diuraphis noxia (Hemiptera: Aphididae) Transcript Expression of the Biotypes SA1 and SAM Feeding on Various Triticum aestivum Varieties.

The intimate relationship between an aphid and its host is mediated by the composition of the secreted saliva. In the present study, aphid heads were sampled and transcript profiling conducted after aphids were fed on their preference host and transferred to a variety of preference and nonpreference hosts. It was found that the virulent Diuraphis noxia (Kurdjumov) (Hemiptera: Aphididae) biotype SAM was able to selectively up-regulate more transcripts when confronted with feeding on a variety of hosts, than was the case with the less virulent D. noxia biotype SA1, suggesting increased genomic regulation when coping with a stressful environment. Collectively, the observed transcriptomic changes are supported by previous findings that host changes induce significant changes in the proteome of phytophagous hemipterans, unlike in many other entomophagous generalist species. The current data suggest that highly specialized hemipterans may be able to counter plant defenses with inducible salivary transcripts with resulting protein biosynthesis, as demonstrated here.

Towards tradable permits for filamentous green algae pollution.

Water pollution permit systems are challenging to design and implement. Operational systems that has maintained functionality remains few and far between, particularly in developing countries. We present current progress towards developing such a system for nutrient enrichment based water pollution, mainly from commercial agriculture. We applied a production function approach to first estimate the monetary value of the impact of the pollution, which is then used as reference point for establishing a reserve price for pollution permits. The subsequent market making process is explained according to five steps including permit design, terms, conditions and transactional protocol, the monitoring system, piloting and implementation. The monetary value of the impact of pollution was estimated at R1887 per hectare per year, which not only provide a "management budget" for filamentous green algae mitigation strategies in the study area, but also enabled the calculation of a reserve price for filamentous green algae pollution permits, which was estimated between R2.25 and R111 per gram filamentous algae and R8.99 per gram at the preferred state.

The expression of selected genes linked to metabolic homeostasis in obese pansteatitis-suffering Mozambique tilapia, Oreochromis mossambicus (Peters).

The Oreochromis mossambicus (Peters) population inhabiting Lake Loskop, South Africa, is characterized by a high incidence of obesity and pansteatitis. We investigated potential links between the impaired health of Lake Loskop O. mossambicus and the endocrine system by assessing the expression of selected genes associated with the thyroid and adrenal endocrine axes as well as peroxisome proliferator-activated receptor gamma (pparg). Moreover, contaminant-induced thyroid and/or metabolic modulation in Lake Loskop water was evaluated using juvenile O. mossambicus in laboratory exposures. The expression of thyroid hormone receptor alpha (thra) and type 2 deiodinase (dio2) was higher in Lake Loskop O. mossambicus than fish from another population, suggesting a degree of thyroid disruption. The altered gene expression may be a consequence, rather than cause of obesity. Expression of dio2 and pparg was higher in juvenile O. mossambicus exposed to unfiltered compared to filtered lake water, and our data suggest fasting as causative factor. Micro-organism abundance can therefore be a confounding factor in studies applying molecular markers to test for thyroid modulation by environmental waters. Pansteatitis was not a significant source of variance in the expression of any of the genes investigated, suggesting that the disease is not associated with disrupted endocrine signalling.

An ecotoxicological screening tool to prioritise acid mine drainage impacted streams for future restoration.

Streams impacted by acid mine drainage (AMD) typically present water exhibiting low pH and high metal concentrations. These factors result in the environmental degradation of watercourses. The objective of this study was to develop and evaluate an ecotoxicological screening tool (EST) to prioritise future remediation of streams impacted by AMD. The Bloubank stream drainage system in South Africa, served as study area for this purpose. In the initial EST development phase physicochemical variables were assessed while in the second phase, epilithic filamentous green algae biomass (chl-a mg m(-2)), diatoms and filamentous green algae community structures were employed as bioindicators as well as Daphnia magna toxicity assays. Using a weight of evidence approach, the first three sites receiving AMD were critically and seriously modified, followed by site 4 that was modified. Sites 1-3 with EST scores ≤70% were assessed as priority candidates for future restoration.

Assessment of the effect of nanomaterials on sediment-dwelling invertebrate Chironomus tentans larvae.

Studies were conducted to determine the effects of a panel of seven nanomaterials (NMs), namely: α-alumina, γ-alumina, precipitated silica; silica fume, calcined silica fume, colloidal antimony pentoxide (Sb(2)O(5)), and superfine amorphous ferric oxide (Fe(2)O(3)), on sediment dwelling invertebrates Chironomus tentans under controlled laboratory conditions. Percentage survival, enzyme activities, growth development, and DNA fragmentation parameters were studied as acute, biochemical, and physiological toxicities of NMs, respectively. Quantitation of catalase and peroxidase enzyme activity demonstrated that toxicant stress of the NMs increased enzyme activity in a concentration dependent fashion across all treatments. The percentage growth length of the test specimens exposed to different NMs was significantly reduced compared to the negative control while only five concentrations were not in the toxic range, namely; Fe(2)O(3) (5 µg/kg); silica fume (5 µg/kg, 50 µg/kg); Sb(2)O(5) (5 µg/kg) and calcined silica fume (5 µg/kg). Genotoxic stress assessed by use of DNA laddering showed complementary findings to the other ecotoxicological endpoints tested in this study--the percentage survival and growth length inhibition.

The effects of engineered nanoparticles on survival, reproduction, and behaviour of freshwater snail, Physa acuta (Draparnaud, 1805).

Increasing uses of engineered nanoparticles (ENPs) in commercial products and industrial applications has eventually resulted to their releases into atmospheric, terrestrial, and aquatic environments. However, knowledge gaps in ENPs toxicity, fate, and behaviour currently limit our ability to quantify risk assessment of materials with nanoscale dimensions, and therefore, the extent of the resultant environmental impacts remains unknown. In the present study, we evaluated the effects of γ-alumina, α-alumina, modified TiO(2) (M-TiO(2)), and commercial TiO(2) (C-TiO(2)) ENPs on the survival, behaviour, and early life stages of the freshwater snail Physa acuta (Draparnaud). The toxicity evaluation was carried out after spiking commercial sand with ENPs concentrations of 0.005, 0.05, or 0.5 gk g(-1). Our findings suggest that increases of γ-alumina and α-alumina concentrations at sub-lethal level concentrations caused significant reduction in the embryo growth rate and embryo hatchability. In addition, these ENPs induced observable developmental deformities of the embryos. In addition, toxicity evaluations using acute 96-h and chronic 28-d tests showed exposure duration may be a significant factor in ENPs-induced toxicity. Therefore, long-term exposure of aquatic organisms to ENPs - potentially can alter certain ecological populations at different trophic levels - and may compromise the entire aquatic ecological functionality. The percentage hatchlings in test chambers containing 0.5 gk g(-1) γ-alumina and α-alumina concentration was 50% less to those observed in the controls. Our results suggest the embryonic growth and hatchability tests are useful endpoints in chronic sediment toxicity tests for determining the toxic thresholds of ENPs in sediment environment. Although no snail mortalities were observed during the static 96-h test containing sediment spiked with different concentrations of M-TiO(2), C-TiO(2), γ-alumina and α-alumina - the antioxidant enzymatic assay results indicated a significant change in antioxidant levels which altered peroxidation at 0.05 or 0.5 gk g(-1)concentrations for both γ-alumina and α-alumina.

Phytobenthos and phytoplankton community changes upon exposure to a sunflower oil spill in a South African protected freshwater wetland.

The occurrence of a sunflower oil spill in 2007 in the Con Joubert Bird Sanctuary freshwater wetland, South Africa, inhibited the growth of sensitive phytoplankton species and promoted that of tolerant species. The algal divisions Chlorophyta and Euglenophyta were well represented in the sunflower oil contaminated water, especially the species Euglena sociabilis, Phacus pleuronectes and Chlamydomonas africana. Young and mature resting zygotes of Chlamydomonas africana were recorded in high abundance at all the sunflower oil contaminated sampling sites. The phytobenthos diversity and abundance were significantly suppressed and negatively associated with low Dissolved Oxygen concentrations and the negative redox potential of the bottom sediment. At the intracellular level, phytoplankton chlorophyll a and b concentrations as physiological variables were more sensitive indicators of the adverse effects of sunflower oil than the 72 h Selenastrum capricornutum algal bioassay conducted.

Limited endosymbiont variation in Diuraphis noxia (Hemiptera: Aphididae) biotypes from the United States and South Africa.

Symbiosis allows an insect access to imbalanced food sources on which other organisms cannot survive. A bacterial endosymbiont, Buchnera aphidicola, gives aphids the ability to feed on phloem depleted of certain essential amino acids by producing those required. Pseudogenes and lower plasmid copy numbers of essential amino acid genes in B. aphidicola, endosymbiont of the Russian wheat aphid, Diuraphis noxia (Kurdjumov) (Hemiptera: Aphididae), suggest that this symbiotic relationship is degenerating. The complete endosymbiont assemblages, copy numbers of plasmids (important in essential amino acid production), and sequence variation in B. aphidicola, from 10 Russian wheat aphid biotypes, were investigated. B. aphidicola was found to be monosymbiotic in the Russian wheat aphid biotypes and other Diuraphis species examined. An insert, occurring in an inverted repeat region on the leucine plasmid, was the only variation found in the approximately 10-kb B. aphidicola sequence analyzed from each Russian wheat aphid biotype. This inverted repeat was shown previously to be conserved within the family Aphididae. The insert occurred in B. aphidicola sequences isolated from four Russian wheat aphid biotypes. Copy numbers of the leucine plasmid differ between the South African and U.S. biotypes and were similar to previously reported values for biotypes from the same geographic regions. These results suggest that B. aphidicola may still contribute to Russian wheat aphid fitness when the aphid feeds on a variety of hosts.

Responses of phytoplankton upon exposure to a mixture of acid mine drainage and high levels of nutrient pollution in Lake Loskop, South Africa.

The relationships between water quality and the phytoplankton community within Lake Loskop were studied during the late summer and autumn of 2008 to evaluate the impacts of acid mine drainage and high nutrient concentrations. The higher concentrations of metal ions and sulphate had adverse effects on certain phytoplankton species in the inflowing riverine zone of Lake Loskop, in comparison to the reference site in the lacustrine zone of the lake, which was dominated by the larger and slower growing late summer species of Coelastrum reticulum Nägeli, Straurastrum anatinum Meyen ex Ralfs and Ceratium hirundinella Müller. The high nutrient concentrations (nitrogen: 17 mg l(-1) and orthophosphate: 0.7 mg l(-1)) during the mid-summer peak of the rainy season were associated with the development of a bloom of the cyanobacterium Microcystis. Water quality data associated with the development of the Microcystis bloom suggest that the aquatic system of Lake Loskop has now entered an alternate, hypertrophic regime. This change overshadowed the adverse effects of high concentrations of heavy metal ions and low pH. Throughout this study, the reference site in the lacustrine zone of Lake Loskop had lower concentrations of metal ions and sulphate, and higher pH values. The response of phytoplankton bioassays on integrated water samples from the different sampling sites did provide potential answers to the reasons for the absence of the algal group Chlorophyceae in the phytoplankton community structure in the riverine zone of the lake.

Biological and chemical evaluation of sewage water pollution in the Rietvlei nature reserve wetland area, South Africa.

Macroinvertebrate communities in Rietvlei nature reserve wetland area and their relationship with water quality were studied with the aim to evaluate their use as potential indicators of pollution. Sampling locations were selected to include outlets from sewage effluent, agricultural and informal residential runoff. A large increase in nutrient concentrations was observed downstream from discharged treated sewage with an associated decrease in species richness. Bioassays performed included: Daphnia magna, Hydra attenuate, Lactuca sativa, Allium cepa and Pyxicephalus adspersus. The highest percentage of lethality response to a screen (100% concentration) of sampled wetland water by test specimens were observed at the point source input of the Hartbeespoort treated sewage plant. Data generated from the AUSRIVAS method and multitrophic level bioassays revealed the deterioration of the wetland possibly due to factors such as increasing urbanization, industrialization, agriculture runoff and rapid human settlement in the Hennops River catchment area and its principal tributaries.

Genetic diversity of African maize inbred lines revealed by SSR markers.

Knowledge of genetic diversity (GD) and relationships among maize inbred lines is indispensable in a breeding program. Our objectives were to (1) investigate the level of genetic diversity among maize inbred lines and (2) assess their genetic structures by applying simple sequence repeat (SSR) markers. Fifty-six highland and mid-altitude maize inbred lines obtained from CIMMYT programs in Ethiopia and Zimbabwe were genotyped using 27 SSR loci. All of the genotypes studied could unequivocally be distinguished with the combination of the SSRs used. In total, 104 SSR alleles were identified, with a mean of 3.85 alleles per locus. The average polymorphism information content (PIC) was 0.58. GD expressed as Euclidean distance, varied from 0.28 to 0.73 with an average of 0.59. Cluster analysis using unweighted pair group method with arithmetic average (UPGMA) suggested five groups among the inbred lines. Most of the inbred lines adapted to the highlands and the mid-altitudes were positioned in different clusters with a few discrepancies. The pattern of groupings of the inbred lines was mostly consistent with available pedigree information. The variability detected using SSR markers could potentially contribute towards effective utilization of the inbred lines for the exploitation of heterosis and formation of genetically diverse source populations in Ethiopian maize improvement programs.

Using a battery of bioassays, benthic phytoplankton and the AUSRIVAS method to monitor long-term coal tar contaminated sediment in the Cache la Poudre River, Colorado.

This survey provides information on sediment toxicity and structural characteristics of the macroinvertebrates and benthic phytoplankton at 10 locations in the Cache la Poudre River after long-term exposure to coal tar residue. The application of the Australian river bioassessment system (AUSRIVAS) as well as a biotest battery was used to evaluate the river 'health' condition. Coal tar is a dense nonaqueous-phase liquid of significant environmental concern due to its toxicity and persistence in the subsurface. Organisms like Selenastrum capricornutum, Daphnia magna and Chironomus tentans, representing different complexities in the biosphere, were selected as test systems for ecotoxicological studies. The results obtained in this study indicate that a biotest battery, macroinvertebrate and benthic phytoplankton communities are in principle suitable biological tools for evaluation of toxic oil and coal-derived substances in long-term contaminated river sediment.

Cloning and sequence analysis of the endopolygalacturonase gene from the pitch canker fungus, Fusarium circinatum.

The fungus Fusarium circinatum causes pitch canker disease on mature pine trees and root rot and damping-off of pine seedlings. Endopolygalacturonases (endoPGs) play a major role during penetration of plants by fungi. Digestion of the pectic polysaccharides in the plant primary cell walls is one of the earliest functions of endoPGs during infection. The research objective was to clone and characterize the gene encoding endopolygalacturonase in F. circinatum. A 970-bp DNA fragment was cloned by using degenerate PCR amplification from F. circinatum DNA. Sequence data for this fragment were used to design specific primers for use in genome walking to amplify and sequence the remaining portion of the F. circinatum endoPG gene (Fcpg). The amino acid sequence predicted from this gene showed 90% and 87% similarity to Fusarium oxysporum and Fusarium moniliforme endoPGs, respectively.

Induction of Pyrophosphate-Dependent Phosphofructokinase in Watermelon (Citrullus lanatus) Cotyledons Coincides with Insufficient Cytosolic D-Fructose-1,6-Bisphosphate 1-Phosphohydrolase to Sustain Gluconeogenesis.

During germination of Citrullus lanatus, pyrophosphate-dependent phosphofructokinase (PFP) activity is induced. The peak of PFP activity coincides with the maximum gluconeogenic flux and high fructose-2,6-bisphosphate (Fru-2,6-P2) concentrations. Determination of cytosolic fructose-1,6 bisphosphatase (FBPase) activity in crude extracts is unreliable because of the high PFP activity. The FBPase activity, after correction for the contaminating PFP, is only one-third of the PFP activity. Purified cytosolic FBPase is inhibited by Fru-2,6-P2. The low cytosolic FBPase activity and high Fru-2,6-P2 most probably result in inadequate in vivo activity to catalyze the observed gluconeogenic flux. The total PFP activity is sufficient to catalyze the required carbon flux.

Pyrophosphate Dependent Phosphofructokinase of Citrullus lanatus: Molecular Forms and Expression of Subunits.

During germination and seedling establishment, the total pyrophosphate-dependent phosphofructokinase (PFP) activity in the cotyledons increases. Two types of subunits with molecular weights of 68 (alpha-subunit) and 65 (beta-subunit) kilodaltons are present. The increase in activity coincides with an approximately 10-fold increase in beta-subunit and twofold increase in alpha-subunit content. Different isoforms of PFP are present at all stages of incubation, but the ratio between the isoforms significantly changes. A linear relationship exists between the ratio of the two PFP subunits and the ratio of the two isoforms of the enzyme. The more anionic (peak 2) isoform of the enzyme apparently is favored by a high ratio of total beta-subunit to alpha-subunit content. The beta- to alpha-subunit ratio of the peak 2 isoform is also approximately fivefold higher than that of the peak 1 (less anionic) isoform. It is evident that the two subunits are not coordinately expressed and the level of expression of each subunit appears to be the primary factor determining the molecular form in which the enzyme is present. In some tissues, only the 65 kilodalton polypeptide is expressed in large amounts. The peak 1 isoform has a higher affinity for pyrophosphate than the peak 2 isoform, while the affinity for fructose-6-phosphate is similar. Both molecular forms are activated by fructose-2,6-bisphosphate.

Control of Pyrophosphated-Fructose-6-Phosphate 1-Phosphotransferase Activity in the Cotyledons of Citrullus lanatus.

After initiation of radicle elongation, the pyrophosphate:d-fructose-6-phosphate 1-phosphotransferase (PFP) activity sharply increases in the cotyledons of Citrullus lanatus. Removal of the radicle early during incubation prevents the increase in PFP activity in the cotyledons evident in the control. Removal of the radicle at any stage after germination results in a decrease in PFP activity in the cotyledons. Application of kinetin (0.5 micromolar) or 2-chlorophosphonic acid (0.1 micromolar) to isolated cotyledons replaces the effect of the radicle. Gibberellic acid (0.09 micromolar GA(3)) also partially mimics the presence of the radicle. Anaerobic conditions, as well as cycloheximide application (0.18 micromolar) to intact embryos or to kinetin and ethrel treated isolated cotyledons prevent the increase in PFP activity evident in the control.