Increased health risk assessment in different vegetables grown under untreated sewerage irrigation regime due to higher heavy metals accumulation.

Heavy metals are environmental pollutants and carcinogenic for human health if ingested. In developing countries, including Pakistan, untreated sewerage water is one of the major sources of irrigation for vegetable production in the vicinities of urban areas which might be toxic to human health due to heavy metals contamination. The present study was conducted to investigate the uptake of heavy metals by sewage water application and its impact on human health. The experiment consisted of five vegetable crops (Raphanus sativus L, Daucus carota, Brassica rapa, Spinacia oleracea, and Trigonella foenum-graecum L.) and two irrigation sources (clean water irrigation and sewage water irrigation). Each treatment was three time replicated for all five vegetables, and standard agronomic practices were applied. The results demonstrated that shoot and root growth in radish, carrot, turnip, spinach, and fenugreek was enhanced significantly with sewerage water, probably due to enhanced organic matter. However, pithiness was observed in the root of radish under sewerage water treatment. Very high concentrations of Cd, up to 7.08 ppm in turnip roots while up to 5.10 ppm in fenugreek shoot, were observed, and other vegetables also contained higher concentrations of Cd. Zn concentrations in the edible parts of carrot (control (C) = 129.17 ppm, sewerage (S) = 164.10 ppm), radish (C = 173.73 ppm, S = 253.03), turnip (C = 109.77 ppm, S = 149.67 ppm), and fenugreek (C = 131.87 ppm, S = 186.36 ppm) were increased by sewerage water treatment but a decrease in Zn concentration in spinach (C = 262.17 ppm, S = 226.97 ppm) was observed. Fe concentration in edible parts of carrot (C = 888.00 ppm, S = 524.80 ppm), radish (C = 139.69 ppm, S = 123.60 ppm), turnip (C = 195.00 ppm, S = 121.37 ppm), and fenugreek (C = 1054.93 ppm, S = 461.77 ppm) were also decreased by sewerage water treatment while spinach leaves had accumulated higher Fe (C = 1560.33 ppm, S = 1682.67 ppm) in sewerage water treatment. The highest bioaccumulation factor value was 4.17 for Cd in carrots irrigated with sewerage water. The maximum value of bioconcentration factor was 3.11 for Cd in turnip under control, and the highest value of translocation factor was 4.82 in fenugreek irrigated with sewerage water. Daily intake of metals and health risk index (HRI) calculation indicated that HRI for Cd was more than 1, suggesting toxicity in these vegetables while HRI for Fe and Zn is still under safe limit. Correlation analysis among different traits of all vegetables under both treatments revealed valuable information for selecting traits in the next crop breeding programs. It is concluded that untreated sewerage-irrigated vegetables, highly contaminated with Cd, are potentially toxic for human consumption and should be banned in Pakistan. Furthermore, it is suggested that the sewerage water should be treated to eliminate toxic compounds, particularly Cd, before irrigation usage and non-edible/phytoremediation crops might be grown in contaminated soils.

Nitrogen optimization coupled with alternate wetting and drying practice enhances rhizospheric nitrifier and denitrifier abundance and rice productivity.

Optimizing nitrogen (N) fertilization without sacrificing grain yield is a major concern of rice production system because most of the applied N has been depleted from the soil and creating environmental consequences. Hence, limited information is available about nutrient management (NM) performance at a specific site under alternate wetting and drying (AWD) irrigation compared to conventional permanent flooding (PF). We aimed to inquire about the performance of NM practices compared to the farmer's fertilizer practice (FFP) under PF and AWD on rhizospheric nitrifier and denitrifier abundance, rice yield, plant growth, and photosynthetic parameters. Two improved NM practices; nutrient management by pig manure (NMPM); 40% chemical N replaced by pig manure (organic N), and nutrient management by organic slow-release fertilizer (NMSR); 40% chemical N replaced by organic slow-release N were compared. The results showed an increased total grain yield (16.06%) during AWD compared to PF. Compared to conventional FFP, NMPM, and NMSR significantly increased the yields by 53.84 and 29.67%, respectively, during AWD. Meanwhile, PF prompted a yield increase of 45.07 and 28.75% for NMPM and NMSR, respectively, (p < 0.05) compared to FFP. Besides, a significant correlation was observed between grain yield and nitrogen content during AWD (R 2 = 0.58, p < 0.01), but no significant correlation was observed during PF. The NMPM contributed to photosynthetic attributes and the relative chlorophyll content under both watering events. Moreover, relatively higher abundances of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) were observed during AWD, and the highest value was found after the late panicle stage. Our results suggest that the AWD-NMPM model is the best option to stimulate nitrifier and denitrifier gene abundance and promote rice production.

RNAi-Mediated Silencing of Putative Halloween Gene Phantom Affects the Performance of Rice Striped Stem Borer, Chilo suppressalis.

The physiological and biochemical characterization of the "Halloween" genes has fundamental importance in the biosynthesis pathway of ecdysteroids. These genes were found to catalyze the final phases of ecdysteroid biosynthesis from dietary cholesterol to the molting hormone 20-hydroxyecdysone. We report the characterization of the Cs-Phm in a major insect pest in agriculture, the rice striped stem borer, Chilo suppressalis (C. suppressalis). A full-length transcript of Cs-Phm was amplified with an open reading frame (ORF) of 478 amino acids through 5' and 3' RACE. Cs-Phm shows five insect-conserved P450 motifs: Helix-C, Helix-I, Helix-K, PERF, and heme-binding motifs. Phylogenetic analysis clearly shows high similarity to Lepidoptera and evolutionary conservation in insects. The relative spatial and temporal transcript profile shows that Cs-Phm is highly expressed in the prothoracic glands and appears throughout the larval development, but with low expression at the start of the larval instar. It seems to peak in 3-4 days and decreases again before the larvae molt. Double-stranded RNA (dsRNA) injection of Cs-Phm at the larval stage efficiently knocked down the target gene and decreased its expression level. The dsRNA-treated group showed significantly decreased ecdysteroid titers, which leads to delayed larval development and higher larval mortality. Negative effects of larval development were rescued by treating 20E in the dsRNA-treated group. Thus, in conclusion, our results suggest that Cs-Phm is functionally conserved in C. suppressalis and encodes functional CYP that contributes to the biogenesis of 20E.

Distribution of polyphenolic compounds, antioxidant potential, and free amino acids in Ziziphus fruits extract; a study for determining the influence of wider geography.

Ziziphus fruits have attracted much attention within the field of medicine due to their high potential against central nervous system disorders. Abundance of secondary metabolites and their composition is key to the pharmaceutical potential and commercial qualities of plants. The in vitro antioxidant activities of Ziziphus nummularia (Burm. f.) and Ziziphus oxyphylla Edgew fruit extract were analyzed using 2,2-diphenil-1-pycrilhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) free radical scavenging assay methods. Phenolic profiles were explored using high-performance liquid chromatography-diode array detector (HPLC-DAD). The result revealed high concentration of polyphenols and their antioxidant potential. In Z. nummularia, the total phenolic content (TPC) (80.270 ± 0.422 µg/ml), DPPH (62.03 ± 0.98 µg/ml), ABTS (66.32 ± 0.73 µg/ml), and TFC (90.683 ± 0.274 µg/ml) were recorded. However, in Z. oxyphylla, DPPH and ABTS values were 60.66 ± 0.56 µg/ml and 61.55 ± 0.77 µg/ml, respectively, indicative of the impacts of climate and soil nutrients. The overall screening of phytochemicals revealed that both the Ziziphus species contain diverse bioactive compounds, including spinacetine-3-O-(2 feruloyl glucopyranosyl)-glucopyranoside, kaempferol-3-O-glucoside-7-O-glucoside, and caffeic acid; p-hydroxybenzoyl hexose, p-coumaric acid, salicylic acid, and ellagic acid pentoxide. Additionally, the highest concentrated amino acid noted was of Lue 0.19 g/100 g with 596.00 retention time (RT), followed by Thr>Ale>Isl>Phya>Val in Z. nummularia. Similarly, the highest concentration of Lue amino acid was recorded as 0.18/100 g with 564.52 RT followed by Pr>Thr>Ale>Lue>Isl>Phya>Val in all genotypes of Z. oxyphylla. Reporting of polyphenols rich and stable species along with identification of favorable regions of cultivation for amino acid, polyphenols, and higher antioxidant potential may lead the way for the identification of elite clones of the species as well as may result in new drug discovery.

Medical Staff Work Burnout and Willingness to Work during COVID-19 Pandemic Situation in Pakistan.

Medical staff is vital for helping society through a health crisis, human-made or natural disaster, and pandemic. This study aims to investigate the medical staff's work-related burnout during Covid-19 and their willingness to work when they are most needed. The cross-sectional design was used, and an online survey was conducted through snowball sampling. Sample comprised on 250 participants (male = 89 & female= 161). The study's inclusion criteria were that only those medical staff of different hospitals was approached to collect data performed inwards isolated for Covid-19 treatment corona isolation wards. Maslach burnout inventory (MBI-HSS) and willingness to work (WTW) tools were used to collect data. Descriptive and Partial least square analysis was utilized to evaluate the relationships. The Coefficient of determination or R-Square value was 0.299, which means 29.9% or 30% of the work burnout variation was due to the impact of emotional exhaustion and personal accomplishment. Perceived danger, Role Competence, Self-Efficacy, and Sense of duty significantly impacted the willingness to work. Despite the workload and perceived risk, 42.6% of participants agreed to work if their department had to need their services, while 55.2% of participants agreed to work whether their department asked them or not. Government and hospital management should adopt a proactive and positive response during the pandemic to eradicate the employee stress and adopt adequate steps to improve the willingness to work with medical staff.

Inter-Parental Conflict's Persistent Effects on Adolescent Psychological Distress, Adjustment Issues, and Suicidal Ideation During the COVID-19 Lockdown.

The current study sought to ascertain the impact of inter-parent conflicts on teenage psychological distress, social and academic adjustment and examine the suicide ideation during the COVID-19. The results found to be alarming as 22% of the individuals displayed suicidal tendencies, with 9% having attempted suicide once, 4.6% having tried suicide twice, and 11% stating that they were likely to do so again. Therefore, the media and the government might host awareness programs and counseling initiatives to promote mental health and prevent suicidal behavior. Moreover, parents may be educated on community level, about the effect of inter-parental arguments on the mental health of their children.

Genome-Wide Analysis of Codon Usage Patterns of SARS-CoV-2 Virus Reveals Global Heterogeneity of COVID-19.

The ongoing outbreak of coronavirus disease COVID-19 is significantly implicated by global heterogeneity in the genome organization of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The causative agents of global heterogeneity in the whole genome of SARS-CoV-2 are not well characterized due to the lack of comparative study of a large enough sample size from around the globe to reduce the standard deviation to the acceptable margin of error. To better understand the SARS-CoV-2 genome architecture, we have performed a comprehensive analysis of codon usage bias of sixty (60) strains to get a snapshot of its global heterogeneity. Our study shows a relatively low codon usage bias in the SARS-CoV-2 viral genome globally, with nearly all the over-preferred codons' A.U. ended. We concluded that the SARS-CoV-2 genome is primarily shaped by mutation pressure; however, marginal selection pressure cannot be overlooked. Within the A/U rich virus genomes of SARS-CoV-2, the standard deviation in G.C. (42.91% ± 5.84%) and the GC3 value (30.14% ± 6.93%) points towards global heterogeneity of the virus. Several SARS-CoV-2 viral strains were originated from different viral lineages at the exact geographic location also supports this fact. Taking all together, these findings suggest that the general root ancestry of the global genomes are different with different genome's level adaptation to host. This research may provide new insights into the codon patterns, host adaptation, and global heterogeneity of SARS-CoV-2.

Phenotypic characterization of exotic tomato germplasm: An excellent breeding resource.

Tomato production in Pakistan faces significant problems of low yields due to various biotic and abiotic stresses primarily because of a narrow genetic base of the cultivars being used. Therefore, Introduction and evaluation of the exotic tomato germplasm has become necessary to acquire elite material to develop future breeding programs. To this end, the present study was conducted for the phenotypic characterization of twenty exotic tomato genotypes along with two locally grown cultivars in semi-arid subtropical climate. Data were collected for morphological, fruit quality and fruit yield traits. A significant (p<0.05) phenotypic variation was observed for all the studied traits. Maximum yield was obtained from "Rober" i.e., 1508.31 g per plant. The maximum shelf life was observed in the Cromco, with the least weight loss (2.45%) and loss in the firmness of fruit (22.61%) in 4 days. Correlation analyses revealed a strong genetic association among morphological and yield related traits. High estimates of the heritability (ranged from 79.77% to 95.01% for different traits), along with a high genetic advance (up to 34%) showed the potential usefulness of these traits and genotypes to develop breeding programs to improve the tomato yield and fruit quality.

Comparative study of the genetic basis of nitrogen use efficiency in wild and cultivated barley.

To curb the increasing demand for nitrogenous fertilizers, it is imperative to develop new cultivars with comparatively greater nitrogen use efficiency (NUE). Nonetheless, so far very meager information is available concerning the variances among barley (Hordeum vulgare L.) varieties for their response to nitrogen deprivation. The current study was carried out to explore the potential of barley genotypes for higher NUE. A hydroponic experiment was conducted at seedling stage to compare the performance of four barley genotypes, ZD9 and XZ149 (with higher NUE) and HXRL and XZ56 (with lower NUE) in response to low (0.1 mM) and normal nitrogen (2 mM) levels. Under low N, all the genotypes expressed less number of tillers, decreased soluble proteins, chlorophyll and N concentrations in both roots and shoots, in comparison with normal N supply. However, significant differences were found among the genotypes. The genotypes with high NUE (ZD9 and XZ149) showed higher N concentration, increased number of tillers, improved chlorophyll and soluble proteins in both roots and shoots as compared to the inefficient ones (HXRL and XZ56). Furthermore, nitrate transporter gene (NRT2.1) showed higher expression under low N, both in roots and leaves of N efficient genotypes, as compared to the N inefficient ones. However, N assimilatory genes (GS1 and GS2) showed higher expression under normal and low N level, in leaves and roots respectively. The outcome of the study revealed that genotypes with higher NUE (ZD9 and XZ149) performed better under reduced N supply, and may require relatively less N fertilizer for normal growth and development, as compared to those with lower NUE. The study also revealed a time-specific expression pattern of studied genes, indicating the duration of low N stress. The current study suggested that future work must involve the time course as a key factor while studying expression patterns of these genes to better understand the genetic basis of low-N tolerance.

Inoculation of Nicotiana tabacum with recombinant potato virus X induces RNA interference in the solenopsis mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae).

OBJECTIVES: The chitin synthase 1 (CHS1) gene in Phenacoccus solenopsis (PsCHS1) was evaluated as a potential target of RNA interference (RNAi) by using Potato virus X (PVX) as a vector (recombinant PVX) for expressing RNAi triggering elements in Nicotiana tabacum L. RESULTS: RT-PCR analysis confirmed the expression of PsCHS1 in N. tabacum inoculated with recombinant-PVX-PsCHS1 (treated). RT- and multiplex-PCR further showed a reduction in mRNA levels of the target gene in mealybugs feeding on treated plants. Mortality in parent adults and emerging nymphs (21 and 29%) exposed to the treated plants was significantly higher (P < 0.05) than those exposed to uninoculated (-ve control) or inoculated with non-recombinant PVX (PVX-control). The number of surviving adults and the combined number of adults and nymphs (47 and 60%) was significantly (P < 0.05) lower on the treated plants than the -ve (76%) or PVX (74%) control. The visual observations verified the physical deformities in mealybugs exposed to the treated plants. CONCLUSION: chitin synthase 1 is a potential RNAi target in P. solenopsis and the recombinant PVX can be used as a tool to evaluate candidate RNAi triggering elements in plants.

Use of recombinant tobacco mosaic virus to achieve RNA interference in plants against the citrus mealybug, Planococcus citri (Hemiptera: Pseudococcidae).

The citrus mealybug, Planococcus citri, is an important plant pest with a very broad plant host range. P. citri is a phloem feeder and loss of plant vigor and stunting are characteristic symptoms induced on a range of host plants, but P. citri also reduces fruit quality and causes fruit drop leading to significant yield reductions. Better strategies for managing this pest are greatly needed. RNA interference (RNAi) is an emerging tool for functional genomics studies and is being investigated as a practical tool for highly targeted insect control. Here we investigated whether RNAi effects can be induced in P. citri and whether candidate mRNAs could be identified as possible targets for RNAi-based P. citri control. RNAi effects were induced in P. citri, as demonstrated by specific target reductions of P. citri actin, chitin synthase 1 and V-ATPase mRNAs after injection of the corresponding specific double-stranded RNA inducers. We also used recombinant Tobacco mosaic virus (TMV) to express these RNAi effectors in Nicotiana benthamiana plants. We found that P. citri showed lower fecundity and pronounced death of crawlers after feeding on recombinant TMV-infected plants. Taken together, our data show that actin, chitin synthase 1 and V-ATPase mRNAs are potential targets for RNAi against P. citri, and that recombinant TMV is an effective tool for evaluating candidate RNAi effectors in plants.