Bacillus safensis filtrate-based ZnO nanoparticles control black heart rot disease of apricot fruits by maintaining its soluble sugars and carotenoids.

Black heart rot is a serious disease of apricot and it has been reported to be caused by Alternaria solani, around the world. The present research was designed to control this disastrous disease using zinc oxide nanoparticles (b-ZnO NPs). These NPs were synthesized in the filtrate of a useful bacterium (Bacillus safensis) and applied to control black heart rot of apricot. After synthesis, the reduction of b-ZnO NPs was confirmed by UV-visible spectroscopy, at 330 nm. Fourier transform infrared (FTIR) spectra ensured the presence of multiple functional groups (alcohols, phenols, carboxylic acids, nitro compounds and amines) on the surface of b-ZnO NPs. X-Ray diffraction (XRD) analysis elucidated their average size (18 nm) while scanning electron microscopy (SEM) micrograph described the spherical shape of b-ZnO NPs. The synthesized b-ZnO NPs were applied in four different concentrations (0.25 mg/ml, 0.50 mg/ml, 0.75 mg/ml, 1.0 mg/ml) under both in vitro and in vivo conditions. These NPs were very efficient in inhibiting mycelial growth (85.1%) of A. solani at 0.75 mg/ml concentration of NPs, in vitro. Same concentration also performed best, in vivo, and significantly reduced disease incidence (by 67%) on self-inoculated apricot fruit. Apart from this, application of b-ZnO NPs helped apricot fruit to maintain its quality under fungal-stress conditions. The decay of apricot fruit was reduced and they maintained greater firmness and higher weight. Moreover, b-ZnO NPs treated fruits controlled black heart rot disease by maintaining higher contents of ascorbic acid, soluble sugars and carotenoids. These b-ZnO NPs were produced in powder form for their easy carriage to the farmers' fields.

Efficacy of organic-based carrier material for plant beneficial rhizobacteria application in okra under normal and salt-affected soil conditions.

AIMS: Plant beneficial rhizobacteria (PBR) improve salt tolerance and plant yield in vegetable plants by producing 1-aminocyclopropane-1-carboxylate-deaminase, indole-3-acetic acid and phosphate solubilization. Organic-based carrier material is needed to ensure the PBR's uniform application, distribution, survival and functioning in a variety of fields. The PBR also use carbon present in the carrier as food and energy source. The selection of a suitable organic-based carrier material for the application of the PBR in normal and saline soils always has received less attention. The current study compared the PBR suitability of different organic-based carrier materials (biochar, biogas residues [BGRs] and coconut powder) and evaluated their effects on okra productivity under normal and saline soil conditions. METHODS AND RESULTS: In a pot experiment, the PBR strain Bacillus sp. MR-1/2 (accession number, MG548383) was applied with/or without organic-based carrier materials to okra grown in three different soils: S1 (EC 1.0 dS m-1 ), S2 (EC 3.0 dS m-1 ) and S3 (EC 5.0 dS m-1 ). The experiment was set up in a completely randomized design with five replicates in factorial arrangement. Results indicated that in soil S1, PBR + BGR increased the number of pods per plant, plant dry weight and indole compounds by 64%, 68% and 17% while reduced the electrolyte leakage (ELL), malonaldehyde (MDA) contents and stress ethylene level by 17%, 55% and 38%, respectively over the PBR application without any carrier. Similarly, in soil S2, the treatment PBR + BGR increased the number of pods by 81%, plant dry weight by 40% and indole compounds by 13% while reduced the ELL by 17%, MDA contents by 50% and stress ethylene by 30% over the PBR alone treatment. In soil S3, PBR + biochar increased the number of pods by 51%, plant dry weight by 62% and indole compounds by 20%, while reduced the ELL by 21%, MDA by 40% and indole compounds by 54% over the PBR alone treatment. CONCLUSIONS: Results concluded that in soil S1 and S2 (normal soils), BGR as carrier for PBR showed best results, while in soil S3, biochar as carrier for PBR resulted in enhanced potassium (K+ ) and calcium (Ca+2 ) uptake and increased the productivity of okra. SIGNIFICANCE AND IMPACT OF STUDY: Response of different carrier materials in supporting PBR under different soil conditions was variable. This study will help in the selection and use of best suitable carrier material for PBR application under different soil conditions. It is recommended that farmer should use BGR as carrier material for PBR application in normal soils while biochar should be used as carrier for the PBR application in saline soil.

Physiological and biochemical characterization of Kalongi (Nigella sativa) against arsenic stress: Implications for human health risk assessment.

Arsenic (As) is a toxic metalloid that exhibits a varying degree of toxicity in plants depending upon the redox status of its species. Elemental arsenic [As(0)] is the least toxic of all the As species, however, under conducive environmental conditions, it can be readily oxidized into toxic forms. The present experiment was designed to evaluate the deleterious effects of As when applied in As(0) form on the morpho-physiological attributes of Kalongi (Nigella sativa). Seeds of N. sativa were sown in soil contaminated with various levels of As (0, 1.875, 3.75, 7.5, 15.0, and 30.0 mg nA(0) kg-1 soil). The results indicated that plant biomass and grain yield of N. sativa were not much affected by various levels of As except at 30 mg nA(0) kg-1 soil. Activities of antioxidant enzymes (SOD, APX, POX, and CAT), phenolic contents, and carotenoids were enhanced in response to the overproduction of H2O2, subsequently inhibiting lipid peroxidation. Arsenic accumulation in different plant organs increased with increasing soil As levels in the given trend root > shoot > leaf > seedpod > seed. Arsenic uptake affected the uptake of other elements (P, Fe, Zn, K, Na, Ca). Adaptive changes in total chlorophyll contents, MDA contents, and antioxidant enzymatic defense mechanism in response to As stress suggest that the N. sativa is tolerant to moderate As stress. Therefore, this crop can be cultivated on moderately As-contaminated soils without any significant risks of economic losses and food chain contamination.

Zinc in soil-plant-human system: A data-analysis review.

Zinc (Zn) plays an important role in the physiology and biochemistry of plants due to its established essentiality and toxicity for living beings at certain Zn concentration i.e., deficient or toxic over the optimum range. Being a vital cofactor of important enzymes, Zn participates in plant metabolic processes therefore, alters the biophysicochemical processes mediated by Zn-related enzymes/proteins. Excess Zn can provoke oxidative damage by enhancing the levels of reactive radicals. Hence, it is imperative to monitor Zn levels and associated biophysicochemical roles, essential or toxic, in the soil-plant interactions. This data-analysis review has critically summarized the recent literature of (i) Zn mobility/phytoavailability in soil (ii) molecular understanding of Zn phytouptake, (iii) uptake and distribution in the plants, (iv) essential roles in plants, (v) phyto-deficiency and phytotoxicity, (vi) detoxification processes to scavenge Zn phytotoxicity inside plants, and (vii) associated health hazards. The review especially compares the essential, deficient and toxic roles of Zn in biophysicochemical and detoxification processes inside the plants. To conclude, this review recommends some Zn-related research perspectives. Overall, this review reveals a thorough representation of Zn bio-geo-physicochemical interactions in soil-plant system using recent data.

Ecotoxicology of Heavy Metal(loid)-Enriched Particulate Matter: Foliar Accumulation by Plants and Health Impacts.

Atmospheric contamination by heavy metal-enriched particulate matter (metal-PM) is highly topical nowadays because of its high persistence and toxic nature. Metal-PMs are emitted to the atmosphere by various natural and anthropogenic activities, the latter being the major source. After being released into the atmosphere, metal-PM can travel over a long distance and can deposit on the buildings, water, soil, and plant canopy. In this way, these metal-PMs can contaminate different parts of the ecosystem. In addition, metal-PMs can be directly inhaled by humans and induce several health effects. Therefore, it is of great importance to understand the fate and behavior of these metal-PMs in the environment. In this review, we highlighted the atmospheric contamination by metal-PMs, possible sources, speciation, transport over a long distance, and deposition on soil, plants, and buildings. This review also describes the foliar deposition and uptake of metal-PMs by plants. Moreover, the inhalation of these metal-PMs by humans and the associated health risks have been critically discussed. Finally, the article proposed some key management strategies and future perspectives along with the summary of the entire review. The abovementioned facts about the biogeochemical behavior of metal-PMs in the ecosystem have been supported with well-summarized tables (total 14) and figures (4), which make this review article highly informative and useful for researchers, scientists, students, policymakers, and the organizations involved in development and management. It is proposed that management strategies should be developed and adapted to cope with atmospheric release and contamination of metal-PM.

Hydrogeochemical investigation of arsenic in drinking water of schools and age dependent risk assessment in Vehari District, Punjab Pakistan: a multivariate analysis.

Globally, a large number of school-aged children is suffering from water-borne diseases, particularly in low-income countries. Arsenic (As) is a hazardous and potentially carcinogenic metal(loid) in drinking water. Nowadays, alarming levels of As have been reported in the groundwater of Vehari District, Punjab Pakistan. In this study, drinking water supplies for high and higher secondary schools were examined in Vehari District. A total of 164 water samples were collected from schools and subjected to heavy metal(loid) analysis (As) and basic water physicochemical parameters. The results were analyzed with respect to sampling area, school type, school education level, sources of sample collection, and the depth of the source. The results revealed that As concentration of water samples in boys' and girls' schools was 12.8 µg/L and 9.2 µg/L, respectively. However, when the As concentration in drinking water was evaluated at the school education level, a notable higher concentration of As was observed in the higher secondary schools than the high schools with an average of 19.5 and 9.7 µg/L, respectively. The risk assessment indices were calculated based on education level and different age groups of the children (primary, elementary, high, and higher secondary). High carcinogenic (cancer risk = 0.001) and non-carcinogenic (hazard quotient = 2.0) risks were noted for the children in higher secondary school. The current findings anticipated that the drinking water of schools in Vehari District did not meet the requirement of the World Health Organization (WHO) drinking water quality guidelines. Safe drinking water is crucial for the development and growth of children. Therefore, it is important for educational authorities to take steps for provision of As free safe drinking water to students and local inhabitants.

Risk assessment and biophysiochemical responses of spinach to foliar application of lead oxide nanoparticles: A multivariate analysis.

Despite extensive research progress in the recent past, the data regarding foliar uptake of heavy metals, associated biophysiochemical changes inside plants and possible health hazards are limited. This study determined the effect of foliar application of lead oxide nanoparticles (PbO-NPs) on lead (Pb) accumulation, physiological and biochemical changes inside spinach plants and associated health risks. A green method was used to prepare PbO-NPs using coconut water. Scanning electron microscopy (SEM) showed the preparation of smooth, unwrinkled, granular and spherical PbO-NPs. Spinach leaves were exposed via foliar application to three concentrations of PbO-NPs (0, 10 and 50 mg/plant). Foliar PbO-NPs application resulted in a significant accumulation of Pb in leaves (42.25 µg g-1), with limited translocation towards root tissues (4.46 µg g-1). This revealed that spinach can accumulate considerable amount of Pb via foliar uptake. Lead accumulation inside spinach caused a significant decrease in pigment contents (38%) and dry weight (67%). After foliar uptake, Pb caused several-fold increase in the activities of catalase and peroxidase. However, foliar PbO-NPs did not induce significant changes in H2O2 production, lipid peroxidation and superoxide dismutase activity. Application of PbO-NPs (50 mg/plant) showed possible health risks (non-carcinogenic) due to ingesting Pb-contaminated leaves of spinach. It is proposed that atmospheric contamination and foliar deposition of metal-PM can seriously affect vegetable growth and can provoke health issues due to ingestion of metal-enriched vegetables. Therefore, atmospheric levels of heavy metals need to be monitored on a regular basis to avoid their food chain contamination and possible human exposure.

Regulation of antioxidant production, ion uptake and productivity in potato (Solanum tuberosum L.) plant inoculated with growth promoting salt tolerant Bacillus strains.

The exchangeable sodium (Na+) in salt affected soils is a major constraint in potassium (K+) availability to plants that disturb ion transport and inhibit plant growth, adversely. Salt tolerant plant growth promoting rhizobacteria (PGPR) may regulate the Na+/K+ efflux and increase K+ uptake by the plant from the soil. Therefore, a pot study was performed to examine the effect of salt tolerant PGPR Bacillus sp. alone and in consortium, on antioxidant enzyme activity, ion uptake and potato (Solanum tuberosum L.) tuber yield in normal and salt affected soils. We observed that Bacillus sp. (strains SR-2-1 and SR-2-1/1) solubilized insoluble phosphorous and produced indole-3-acetic acid while only SR-2-1/1 produced ACC deaminase in culture medium supplemented with various concentrations of NaCl (0-6%). In the pot experiment, the consortium treatment of strains was found to increase relative leaf water contents whereas decreased the electrolyte leakage and antioxidant enzyme activity both in normal and salt affected soils. Similarly, consortium treatment decreased Na+ whereas increased K+, Ca+2, K+/Na+ and Ca+2/Na+ in plant dry matter in both soils. It has been investigated that inoculation of PGPR significantly (p < 0.05) increased plant biomass, number of tubers per plant and tuber weight as compared to un-inoculated plants in both soils. In addition, PGPR inoculation enhanced auxin production in root exudates of young potato plants and bacterial population dynamics in both soils. Na+ ion regulation (R2 = 0.95) and tuber weight (R2 = 0.90) in salt affected soil were significantly correlated with auxin production in the rhizosphere. Results of this study conferred that consortium of Bacillus strains (SR-2-1, SR-2-1/1) enhanced auxin production in the rhizosphere of potato plants and that ultimately regulated antioxidant enzyme production and uptake of Na+, K+ and Ca+2 in potato plants resulted into a higher tuber yield in both normal and salt affected soils.

A multivariate analysis of health risk assessment, phytoremediation potential, and biochemical attributes of Spinacia oleracea exposed to cadmium in the presence of organic amendments under hydroponic conditions.

Cadmium (Cd) phytoremediation potential and its accumulation in edible and nonedible plant tissues is the function of various biochemical processes taking place inside plants. This study assessed the impact of organic ligands on Cd phyto uptake and different biophysiochemical processes of Spinacia oleracea L., and associated health hazards. Plants were exposed to Cd alone and chelated with citric acid (CA) and ethylenediaminetetraacetic acid (EDTA). Results revealed that the effect of Cd on lipid peroxidation, H2O2 production and pigment contents varied greatly with its applied level and the type of organic ligand. Moreover, the effect was more prominent in root tissues than leaf tissues and for high concentrations of Cd and organic ligands. Cadmium accumulation increased by 90 and 74% in roots and leaves, respectively, with increasing Cd levels (25-100 µM). Cadmium exposure at high levels caused lipid peroxidation in roots only. Application of both CA and EDTA slightly diminished Cd toxicity with respect to pigment contents, lipid peroxidation and hydrogen peroxide (H2O2) contents. Hazard quotient (HQ) of Cd was <1.00 for all the treatments. Under nonlinear effect of treatments, multivariate analysis can be an effective tool to trace overall effects/trends.

Foliar uptake of arsenic nanoparticles by spinach: an assessment of physiological and human health risk implications.

Atmospheric contamination by heavy metal(loid)-enriched particulate matter (metal-PM) is highly topical these days because of its high persistence, toxic nature, and health risks. Globally, foliar uptake of metal(loid)s occurs for vegetables/crops grown in the vicinity of industrial or urban areas with a metal-PM-contaminated atmosphere. The current study evaluated the foliar uptake of arsenic (As), accumulation of As in different plant organs, its toxicity (in terms of ROS generation, chlorophyll degradation, and lipid peroxidation), and its defensive mechanism (antioxidant enzymes) in spinach (Spinacia oleracea) after foliar application of As in the form of nanoparticles (As-NPs). The As-NPs were prepared using a chemical method. Results indicate that spinach can absorb As via foliar pathways (0.50 to 0.73 mg/kg in leaves) and can translocate it towards root tissues (0.35 to 0.68 mg/kg). However, health risk assessment parameters showed that the As level in the edible parts of spinach was below the critical limit (hazard quotient < 1). Despite low tissue level, As-NP exposure caused phytotoxicity in terms of a decrease in plant dry biomass (up to 84%) and pigment contents (up to 38%). Furthermore, several-fold higher activities of antioxidant enzymes were observed under metal stress than control. However, no significant variation was observed in the level of hydrogen peroxide (H2O2), which can be its possible transformation to other forms of reactive oxygen species (ROS). It is proposed that As can be absorbed by spinach via foliar pathway and then disturbs the plant metabolism. Therefore, air quality needs to be considered and monitored continuously for the human health risk assessment and quality of vegetables cultivated on polluted soils (roadside and industrial vicinity). Graphical abstract ᅟ.

Effect of organic amendments on cadmium stress to pea: A multivariate comparison of germinating vs young seedlings and younger vs older leaves.

Despite significant recent advancement in research, biogeochemical behavior of heavy metals with respect to their applied form is still topical. Moreover, metal toxicity to plants may vary with their stage of development/maturity. Therefore, this study for the first time evaluated the influence of ethylenediaminetetraacetic acid (EDTA) and citric acid (CA) on cadmium (Cd) accumulation and toxicity to germinating and young pea seedlings as well as in younger and older leaves. The experimental setup of current study consisted of two separate studies. The first study was performed on germinating seedlings grown in a Cd-contaminated sand media. Pea seeds were treated with two levels of Cd (Cd-25 and Cd-100) alone and combined with different levels of EDTA and CA. The second study was carried out in hydroponic solution. The influence of organic amendments on Cd accumulation and toxicity to pea plants was evaluated by determining Cd contents in pea seedlings, H2O2 contents, chlorophyll contents and lipid peroxidation in younger and older leaves. Cadmium stress caused overproduction of H2O2 in roots and leaves of pea seedlings. Cadmium-induced overproduction of H2O2 caused a decrease in the pigment contents and increased lipid peroxidation. Application of EDTA at higher levels (81 and 200µM) increased Cd accumulation by pea plants. However, CA did not affect Cd accumulation by pea. Both EDTA and CA increased Cd-induced H2O2 production and lipid peroxidation. Younger pea leaves showed more sensitivity to Cd stress compared to older leaves. Similarly, Cd toxicity was more pronounced in germinating seedlings than young seedlings. Moreover, Pearson correlation and principal component analysis (PCA) showed very interesting correlations between treatments and stress responses of germination and young seedlings as well as younger and older leaves. Based on multivariate analysis, it is proposed that the Cd toxicity to pea plants greatly vary with its growth stage and the maturity of organs (younger or older leaves).

Virulence determination and molecular features of peach latent mosaic viroid isolates derived from phenotypically different peach leaves: a nucleotide polymorphism in L11 contributes to symptom alteration.

Symptoms of chlorosis along leaf edges (chlorosis-edge), along leaf veins (chlorosis-vein) and yellowing on peach leaves have been observed for a long history in the field, while the pathological factor(s) responsible for these symptoms remained unknown. Peach latent mosaic viroid (PLMVd) was detected in the leaves collected from three unique phenotypic peach trees showing above mentioned symptoms. The obtained PLMVd isolates were subjected to population structure analyses and biological assays to evaluate their pathogenicity on peach seedlings in an effort to elucidate the relationship between the PLMVd and the symptoms observed on peach trees in China. In addition, molecular features of PLMVd isolates were analyzed to obtain some insight into the structure-function relationships of this viroid. The results revealed that the symptoms of chlorosis-edge and yellowing were indeed incited by PLMVd, and a direct link between the nucleotide polymorphisms and the symptoms of yellowing and chlorosis-edge was established, i.e. residue U338 responsible for the yellowish symptom and C338 responsible for the chlorosis-edge symptom. This study provides an additional proof to endorse a previous proposal that PLMVd pathogenicity determinants reside in L11. The illustrative etiology of the disease, visualization of the symptoms progression and identification of the unique single nucleotide polymorphism possibly involved in the symptom induction will significantly increase understanding of the pathogenic mechanisms of PLMVd and will help in designing control strategies for the resulting disease.

Genetic diversity analyses reveal novel recombination events in Grapevine leafroll-associated virus 3 in China.

Grapevine leafroll-associated virus 3 (GLRaV-3) is the most prevalent causal agent of grapevine leafroll disease (GLD). Of the 75 grapevine samples collected from three regions in China, 46.7% and 94.7% of samples tested positive for GLRaV-3 in reverse transcription-PCR (RT-PCR) and reverse transcription nested PCR (RT-nPCR), respectively. The SSCP analysis for the clones of complete CP gene from 16 GLRaV-3 isolates showed that 15 isolates contained one predominant haplotype and one isolate had no predominant haplotype. The sequences of the CP genes showed 89.9-100% identities at the nucleotide level. Phylogenetic analysis of the CP gene sequences revealed the existence of four well defined variant groups, which corresponded to previously reported phylogenetic groups (1, 2, 3, and 5). Two new sub-groups designated as sub-group 1B and sub-group 3B in groups 1 and 3, respectively, were identified in the Chinese GLRaV-3 population. Recombination analyses illustrated that those two new sub-groups (1B and 3B) were emerged as a result of recombination events between variants in groups 1 and 2, and variants in groups 1 and 3, respectively. These results further indicated that the variants in those new sub-groups are viable and evolutionary successful. Recombinants with highly similar coat protein structure to variants of group 1 were abundantly found in the viral population. In addition, these analyses provided evidence about CP gene as one of the recombination hotspots in GLRaV-3 genome. The population genetic parameters of all available CP sequences suggested that the recombinants might have emerged due to population bottlenecks during transmission. The results provide new insights into the variability and evolution of GLRaV-3.

Probe binding to host proteins: a cause for false positive signals in viroid detection by tissue hybridization.

Molecular hybridization assay, especially involving the use of tissues directly, has been developed as a rapid, simple and important technique for plant pathogen detection and/or gene expression analysis on a large scale. In theory, this method relies on the specific binding of a labeled probe to a target nucleotide sequence. However, occasional false positive reactions can pose a problem in its application and the cause is often not well understood. Here, we show that in tissue-printing hybridization to detect Peach latent mosaic viroid (PLMVd) strong signals could arise by interactions between the viroid probe and plant proteins. Such probe-protein interactions made it difficult to show significant correlations between viroid infection and the level of hybridization signals. These results challenge the traditional view that proteins may hamper PCR reactions but have no influence on molecular hybridization. They further demonstrate that such probe-protein interactions in a plant could compromise the quality of molecular hybridization assays for viroid detection. Our results uncovered an important source of false positive reactions in tissue-printing hybridization and suggest that specificity can be improved by removing proteins.