This research was designed to elucidate the role of exopolysaccharides (EPS) producing bacterial strains for the amelioration of drought stress in wheat. Bacterial strains were isolated from a farmer’s field in the arid region of Pakistan. Out of 24 isolated stains, two bacterial strains, Bacillus subtilis (Accession No. MT742976) and Azospirillum brasilense (Accession No. MT742977) were selected, based on their ability to produce EPS and withstand drought stress. Both bacterial strains produced a good amount of EPS and osmolytes and exhibited drought tolerance individually, however, a combination of these strains produced higher amounts of EPS (sugar 6976 µg/g, 731.5 µg/g protein, and 1.1 mg/g uronic acid) and osmolytes (proline 4.4 µg/mg and sugar 79 µg/mg) and significantly changed the level of stress-induced phytohormones (61%, 49% and 30% decrease in Indole Acetic Acid (IAA), Gibberellic Acid (GA), and Cytokinin (CK)) respectively under stress, but an increase of 27.3% in Abscisic acid (ABA) concentration was observed. When inoculated, the combination of these strains improved seed germination, seedling vigor index, and promptness index by 18.2%, 23.7%, and 61.5% respectively under osmotic stress (20% polyethylene glycol, PEG6000). They also promoted plant growth in a pot experiment with an increase of 42.9%, 29.8%, and 33.7% in shoot length, root length, and leaf area, respectively. Physiological attributes of plants were also improved by bacterial inoculation showing an increase of 39.8%, 61.5%, and 45% in chlorophyll a, chlorophyll b, and carotenoid content respectively, as compared to control. Inoculations of bacterial strains also increased the production of osmolytes such asproline, amino acid, sugar, and protein by 30%, 23%, 68%, and 21.7% respectively. Co-inoculation of these strains enhanced the production of antioxidant enzymes such as superoxide dismutase (SOD) by 35.1%, catalase (CAT) by 77.4%, and peroxidase (POD) by 40.7%. Findings of the present research demonstrated that EPS, osmolyte, stress hormones, and antioxidant enzyme-producing bacterial strains impart drought tolerance in wheat and improve its growth, morphological attributes, physiological parameters, osmolytes production, and increase antioxidant enzymes.

In this study, heavy metal phyto-accumulation potential of selected cultivars of two leafy vegetables on irrigation with municipal wastewater and human health risks were investigated. Municipal wastewater chemistry was recorded significantly different from groundwater control and led to the two-fold high enrichment of soil heavy metal contents (Ni, 19.46; Pb, 23.94; Co, 4.68; Cd, 1.4 in mg/kg, respectively). Interactive effects for phyto-accumulation of most heavy metals were also recorded significant at p < 0.001 in four vegetable cultivars. Heatmap revealed higher accumulation of heavy metals (Fe, Zn, Mn, Cu, Pb, Cr, Co) in spinach cultivars than lettuce cultivars creating elevated health risk index (HRI) and hazard index (HI) values for adults and children. Highest HI was recorded for Lahori palak (adults, 1.42; children, 2.58) and lowest for iceberg (adults, 0.04; children, 0.07). The NPK supplementation improved mineral composition of leafy vegetables within safer human health limits in control treatments. However, in municipal wastewater treatments, NPK fertilization decreased heavy metal uptake and phyto-accumulation in S2 (Lahori palak) than remaining vegetable cultivars leading to reduced health risk values. Because of higher heavy metal phyto-accumulation and health risks, cultivation of spinach cultivars must be discouraged in agro-ecologies receiving municipal wastes, whereas lettuce cultivars should be promoted.

The present study aimed to reveal the impact of the stay-green trait in bread wheat under terminal heat stress. Field experiments (early and late sowing; for two consecutive years) were conducted to investigate the influence of terminal heat stress on the morpho-physiological traits in different stay-green types i.e., non-stay-green, moderately non-stay-green, moderately stay-green, and stay-green. In addition, the greenhouse experiment was performed to dissect the stay-green trait in functional stay-green, non-functional stay-green, and non-stay-green genotypes. The results of the field experiments confirmed that genotypes exhibiting the stay-green trait have a significantly high chlorophyll content, normalized difference vegetative index, grain yield, biological yield, kernel weight, and low canopy temperature under control and heat stress conditions. In the greenhouse experiment, functional stay-green and non-functional stay-green genotypes showed a high chlorophyll content and photochemical efficiency, whereas biological yield and grain yield showed a significant relation with the functional stay-green genotype under control and terminal heat stress treatments. The sequencing and expression analysis of chlorophyllide a oxygenase (CaO), light-harvesting complex (Cab), stay-green (SGR), and red chlorophyll catabolite reductase (RCCR) in functional stay-green, non-functional stay-green, and non-stay-green genotypes revealed variations in the exons of CaO and RCCR; and significant difference in the regulation of CaO and Cab at 7 days after anthesis under terminal heat stress. This study confirms that genotypes displaying the stay-green trait can aid wheat breeders to cope with increasing temperature in the impending decades.

Design and implementation of wastewater treatment is inevitable due to toxic effects of wastewater irrigation on crops, soil and human health. Current investigation is the pioneer attempt on full-scale hybrid constructed wetland system (HCWS) built for municipal wastewater treatment from Pakistan. HCWS was comprised of vertical sub-surface flow constructed wetland (VSSF-CW) and five phyto-treatment ponds connected in series. Higher environmental risk was associated with untreated municipal wastewater usage in irrigation as estimated through discharge of metals to recipient soils. Treatment efficiency percentages recorded for HCWS reclaimed water quality parameters were, i.e., EC (56.68), TDS (56.86), alkalinity (39.67), chloride (39.68), sulfate (46.73), Na (28.80), Mn (65.24), Cr (78.07), Ni (81.02), BOD (68.74), total hardness (19.56), Fe (70.09), phosphate (55.40), Pb (80.48), COD (63.64), Mg (17.24), K (60.05), Co (100), Cu (67.73), Zn (59.97), Cd (100), and Ca (21.47) respectively. Wastewater treatment in HCWS was due to aquatic plants [Phragmites australis Cav. Trin. ex Steud., Canna indica L. Typha latifolia L., and Hydrocotyle umbellata L.], microbial activities and substrate based wetland processes. The HCWS treated water was well under irrigation standards and recommended for safer crop production in water scarce regions.

Nitrogen (N) deficiency and heat stress (HS) are major abiotic stresses that affect the quantity and quality of wheat grains. This study was conducted to examine wheat varietal response to RSI and RNDVI at the anthesis stage and their relationship to yield and yield-related traits under variable N supply and terminal heat stress. Twelve wheat varieties were evaluated in 2016–2017 and 2017–2018 at the National Agricultural Research Centre (NARC), Islamabad, Pakistan. The experiment was divided into three sets, i.e., N120 (120 kg N/ha), N60 (60 kg N/ha) and N0 (0 kg N/ha), based on the nitrogen fertilizer application. The physiological and yield-related parameters were recorded. Mean grain yield for all twelve varieties, averaged from two years of data, ranged between 1655.0 and 3890.1 kg/ha. Maximum RSI (0.99), RNDVI (1.03) and GY (3890.9 kg/ha) were recorded for FSD-08, while AARI-11 showed minimum RSI (0.50), RNDVI (0.56) and GY (1396.40 kg/ha). In the present study, mean CTD was lower, at N0 (3.57 °C), followed by N60 (5.07 °C) and N120 (5.47 °C) on average for the two years of data. The strong positive correlation of RSI and RNDVI with grain yield at R² = 0.73 and R² = 0.49 suggest that these parameters can be used as efficient and precise selection criteria for identifying nitrogen-use-efficient wheat varieties under terminal heat-stress conditions. This work will help the researchers to identify and develop nitrogen-use-efficient and thermos-tolerant wheat cultivars by minimizing the negative impacts of heat stress at the anthesis stage.