Cloning of CAT genes in Satsuma mandarin and their expression characteristics in response to environmental stress and arbuscular mycorrhizal fungi.

KEY MESSAGE: CitCAT1 and CitCAT2 were cloned and highly expressed in mature leaves. High temperatures up-regulated CitCAT1 expression, while low temperatures and Diversispora versiformis up-regulated CitCAT2 expression, maintaining a low oxidative damage. Catalase (CAT), a tetrameric heme-containing enzyme, removes hydrogen peroxide (H2O2) to maintain low oxidative damage in plants exposed to environmental stress. This study aimed to clone CAT genes from Citrus sinensis cv. "Oita 4" and analyze their expression patterns in response to environmental stress, exogenous abscisic acid (ABA), and arbuscular mycorrhizal fungal inoculation. Two CAT genes, CitCAT1 (NCBI accession: PP067858) and CitCAT2 (NCBI accession: PP061394) were cloned, and the open reading frames of their proteins were 1479 bp and 1539 bp, respectively, each encoding 492 and 512 amino acids predicted to be localized in the peroxisome, with CitCAT1 being a stable hydrophilic protein and CitCAT2 being an unstable hydrophilic protein. The similarity of their amino acid sequences reached 83.24%, and the two genes were distantly related. Both genes were expressed in stems, leaves, flowers, and fruits, accompanied by the highest expression in mature leaves. In addition, CitCAT1 expression was mainly up-regulated by high temperatures (37 °C), exogenous ABA, and PEG stress within a short period of time, whereas CitCAT2 expression was up-regulated by exogenous ABA and low-temperature (4 °C) stress. Low temperatures (0 °C) for 12 h just up-regulated CitCAT2 expression in Diversispora versiformis-inoculated plants, and D. versiformis inoculation up-regulated CitCAT2 expression, along with lower hydrogen peroxide and malondialdehyde levels in mycorrhizal plants at low temperatures. It is concluded that CitCAT2 has an important role in resistance to low temperatures as well as mycorrhizal enhancement of host resistance to low temperatures.

Insight into the Morphological Properties of Nano-Kaolinite (Nanoscrolls and Nanosheets) on Its Qualification as Delivery Structure of Oxaliplatin: Loading, Release, and Kinetic Studies.

Natural kaolinite underwent advanced morphological-modification processes that involved exfoliation of its layers into separated single nanosheets (KNs) and scrolled nanoparticles as nanotubes (KNTs). Synthetic nanostructures have been characterized as advanced and effective oxaliplatin-medication (OXAP) delivery systems. The morphological-transformation processes resulted in a remarkable enhancement in the loading capacity to 304.9 mg/g (KNs) and 473 mg/g (KNTs) instead of 29.6 mg/g for raw kaolinite. The loading reactions that occurred by KNs and KNTs displayed classic pseudo-first-order kinetics (R2 > 0.90) and conventional Langmuir isotherms (R2 = 0.99). KNTs exhibit a higher active site density (80.8 mg/g) in comparison to KNs (66.3 mg/g) and raw kaolinite (6.5 mg/g). Furthermore, compared to KNs and raw kaolinite, each site on the surface of KNTs may hold up to six molecules of OXAP (n = 5.8), in comparison with five molecules for KNs. This was accomplished by multi-molecular processes, including physical mechanisms considering both the Gaussian energy (<8 KJ/mol) and the loading energy (<40 KJ/mol). The release activity of OXAP from KNs and KNTs exhibits continuous and regulated profiles up to 100 h, either by KNs or KNTs, with substantially faster characteristics for KNTs. Based on the release kinetic investigations, the release processes have non-Fickian transport-release features, indicating cooperative-diffusion and erosion-release mechanisms. The synthesized structures have a significant cytotoxicity impact on HCT-116 cancer cell lines (KNs (71.4% cell viability and 143.6 g/mL IC-50); KNTs (11.3% cell viability and 114.3 g/mL IC-50). Additionally, these carriers dramatically increase OXAP's cytotoxicity (2.04% cell viability, 15.4 g/mL IC-50 (OXAP/KNs); 0.6% cell viability, 4.5 g/mL IC-50 (OXAP/KNTs)).

Polyphenols-Rich Extract of Calotropis procera Alone and in Combination with Trichoderma Culture Filtrate for Biocontrol of Cantaloupe Wilt and Root Rot Fungi.

Fungal diseases have always been a major problem for cantaloupe crops; however, synthetic fungicides are hazardous to humans and the environment. Consequently, a feasible alternative to fungicides without side effects could be by using bio agents and naturally occurring plants with antibacterial potential. This study has achieved a novel procedure for managing wilt and root rot diseases by potentially using Trichoderma sp. culture filtrates in consortium with plant extract of Calotropis procera, Rhizoctonia solani, Fusarium oxysporum, and Pythium ultimum, which were isolated from infected cantaloupe roots with identified root rot symptoms. The antagonistic activity of four Trichoderma isolates and analysis of antibiotics and filtrate enzymes of the most active Trichoderma isolate were determined as well as phytochemical analysis of C. procera plant extract using HPLC-UV. The obtained results showed that all Trichoderma isolates considerably lowered the radial growth of P. ultimum, R. solani, and F. oxysporum in varying degrees. The scanning electron micrographs illustrate the mycoparasitic nature of Trichoderma sp. on F. oxysporum. The phytochemical analysis of C. procera indicated that phenolic contents were the major compounds found in extracts, such as vanillin (46.79%), chlorogenic acid (30.24%), gallic acid (8.06%), and daidzein (3.45%) but including only a low amount of the flavonoid compounds rutin, naringenin, and hesperetin. The Pot experiment's findings showed that cantaloupe was best protected against wilting and root rot diseases when it was treated with both Trichoderma sp. culture filtrates (10%) and C. procera extract of (15 mg/mL), both alone and in combination. This study demonstrates that the application of bio agent Trichoderma spp. filtrate with C. procera phenol extract appears useful for controlling wilting and root rot disease in cantaloupe. This innovative approach could be used as an alternative to chemical fungicide for the control of wilting and rot root diseases.

Endophytic Bacillus vallismortis and Bacillus tequilensis bacteria isolated from medicinal plants enhance phosphorus acquisition and fortify Brassica napus L. vegetative growth and metabolic content.

Phosphorus fertilization imposes critical limitations on crop productivity and soil health. The aim of the present work is to explore the potential of two phosphate solubilizing bacteria (PSB) species in phosphorus supplementation of canola (Brassica napus L.). Out of 38 bacterial isolates obtained from nine medicinal plants, two bacterial strains (20P and 28P) were proved as the most potent for the in-vitro tricalcium phosphate solubilization test. These isolates verified their activity toward different enzymes as nitrogenase and alkaline phosphatase. Also, 20P and 28P gave a high amount of indole-3-acetic acid, 34.16 µg/ml and 35.20 µg/ml, respectively, and were positive for siderophores production as they detected moderate affinity for iron chelation. Molecular identification confirmed that strain 20P was Bacillus vallismortis and strain 28P was Bacillus tequilensis. A pot experiment was conducted to study the effect of four different phosphorus concentrations (0%, 50%, 75%, and 100% P) each alone and/or in combination with B. vallismortis, B. tequilensis, or both bacterial isolates on the vegetative growth and some physiological parameters of canola. The combined treatment of 50% phosphorus + (B. vallismortis + B. tequilensis) was generally the most effective with respect to shoot height, shoot dry mass, leaf area, photosynthetic pigment fractions, total sugar content, and accumulated NPK content. In contrast, the rhizosphere pH reached the minimum value under the same treatment. These findings highlighted the potential use of PSB (B. vallismortis and B. tequilensis) along with phosphorus fertilization as a safe sustainable tactic.

Effect of Biotic Elicitors on the Growth, Antioxidant Activity and Metabolites Accumulation in In Vitro Propagated Shoots of Pueraria tuberosa.

Pueraria tuberosa contains a wide range of bioactive compounds, including polyphenols, alkaloids, and phytosterols, which make it valuable to the pharmaceutical and food industries. Elicitor compounds trigger the defense mechanisms in plants and are widely used to increase the yield of bioactive molecules in in vitro cultures. The present study was conducted to evaluate the effects of different concentrations of biotic elicitors such as yeast extract (YE), pectin (PEC), and alginate (ALG) on growth, antioxidant activity, and metabolite accumulation in in vitro propagated shoots of P. tuberosa. The elicitors applied to shoot cultures of P. tuberosa significantly increased biomass (shoot number, fresh weight, and dry weight), and metabolites such as protein, carbohydrates, chlorophyll, total phenol (TP), and total flavonoid (TF) contents, as well as antioxidant activity compared to untreated control. Biomass, TP, and TF contents, as well as antioxidant activity, were most significant in cultures treated with 100 mg/L PEC. In contrast, chlorophyll, protein, and carbohydrate increased most in cultures treated with 200 mg/L ALG. Application of 100 mg/L of PEC led to the accumulation of high amounts of isoflavonoids including puerarin (220.69 µg/g), daidzin (2935.55 µg/g), genistin (5612 µg/g), daidzein (479.81 µg/g), and biochanin-A (111.511 µg/g) as analyzed by high-performance liquid chromatography (HPLC). Total isoflavonoids content of 100 mg/L PEC treated shoots was obtained as 9359.56 µg/g, 1.68-fold higher than in vitro propagated shoots without elicitors (5573.13 µg/g) and 2.77-fold higher than shoots of the mother plant (3380.17 µg/g). The elicitor concentrations were optimized as 200 mg/L YE, 100 mg/L PEC, and 200 mg/L ALG. Overall, this study showed that the application of different biotic elicitors resulted in better growth, antioxidant activity, and accumulation of metabolites in P. tuberosa, which could lead to obtaining phytopharmaceutical advantages in the future.

Wheat Grains as a Sustainable Source of Protein for Health.

Protein deficiency is recognized among the major global health issues with an underestimation of its importance. Genetic biofortification is a cost-effective and sustainable strategy to overcome global protein malnutrition. This study was designed to focus on protein-dense grains of wheat (Triticum aestivum L.) and identify the genes governing grain protein content (GPC) that improve end-use quality and in turn human health. Genome-wide association was applied using the 90k iSELECT Infinium and 35k Affymetrix arrays with GPC quantified by using a proteomic-based technique in 369 wheat genotypes over three field-year trials. The results showed significant natural variation among bread wheat genotypes that led to detecting 54 significant quantitative trait nucleotides (QTNs) surpassing the false discovery rate (FDR) threshold. These QTNs showed contrasting effects on GPC ranging from -0.50 to +0.54% that can be used for protein content improvement. Further bioinformatics analyses reported that these QTNs are genomically linked with 35 candidate genes showing high expression during grain development. The putative candidate genes have functions in the binding, remobilization, or transport of protein. For instance, the promising QTN AX-94727470 on chromosome 6B increases GPC by +0.47% and is physically located inside the gene TraesCS6B02G384500 annotated as Trehalose 6-phosphate phosphatase (T6P), which can be employed to improve grain protein quality. Our findings are valuable for the enhancement of protein content and end-use quality in one of the major daily food resources that ultimately improve human nutrition.

Pre and postharvest characteristics of Dahlia pinnata var. pinnata, cav. As affected by SiO2 and CaCO3 nanoparticles under two different planting dates.

Agriculture faces many challenges because of climate changes. The nutrients present in nano-sized form improve plant productivity, especially when used at the appropriate planting time. Field experiments were conducted as a factorial experiment for evaluating two planting dates (20th September and 20th October), foliar application with nanoparticles (NPs) including silica nanoparticles (SiO2-NPs) at 1.5 and 3 mM, calcium carbonate nanoparticles (CaCO3-NPs) at 5 and 10 mM and distilled water (control) on pre- and post-harvest characteristics of Dahlia pinnata var. pinnata Cav. The results indicate that the interactions during the late planting time (20th October) and exogenous applications of SiO2-NPs at 1.5 mM or CaCO3-NPs at 10 mM have improved plant growth including plant height, stem diameter, fresh and dry weights of plant, leaf area, inflorescence diameter, inflorescence stalk length, branches number, tuber numbers, inflorescences number on the plant, and the vase life. At the same time, insignificant differences appeared in the interaction during the planting dates and SiO2 or CaCO3 -NPs concentrations on inflorescence stalk diameter, total soluble solids, membrane stability index, maximum increase in fresh weight (FW), and Si and Ca contents. In addition, all exogenous applications of NPs at the late planting time promoted the plant growth characteristics like lignin %, cellulose %, inflorescence water content, change in FW, and total water uptake. Moreover, the controls through the two planting dates recorded the maximum change in water uptake and water loss values. In short, it can be recommended to use SiO2-NPs at 1.5 mM or CaCO3-NPs at 10 mM as a foliar application at the late planting time (20th October) for obtaining the optimum quantitative and qualitative parameters of D. pinnata.

Enhancing Sugarcane Yield and Sugar Quality through Optimal Application of Polymer-Coated Single Super Phosphate and Irrigation Management.

The judicious use of crop input is of prime importance for achieving a considerable output with a low-cost input. A two-year field experimentation was executed to assess the effect of varying polymer-coated single super phosphate (SSP) regimes on the yield and quality of sugarcane under differential water regimes. A two-factor study was executed under a randomized complete block design with a split-plot arrangement. The CPF-249 sugarcane variety was planted during the 2019-2020 period and the 2020-2021 period. The experiment consisted of four levels of polymer-coated SSP, i.e., control, 90, 110, and 130 kg ha-1, and three water regimes, which consisted of a number of irrigations, i.e., 18 irrigations, 15 irrigations, and 12 irrigations. Moreover, the water regimes were kept in the main plot, whereas the polymer-coated supplement was allocated in a subplot and replicated thrice. The data on the yield components and sugar-related traits were recorded during both years of study, and the treatment means were differentiated using an LSD test at a 95% confidence interval. Summating the findings of this study, a significant variation was revealed under the subject levels of both factors. Statistically, a 110 kg ha-1 polymer-coated SSP dose, along with 18 irrigations, declared the highest millable canes, stripped cane yield, and unstripped cane yield, followed by the 130 kg ha-1 treatment. Additionally, the highest pol% and cane sugar recovery % were recorded under 12 irrigations along with 130 kg ha-1 during both years. Similarly, the °Brix value was also significantly affected by 12 irrigations when 110 kg ha-1 of polymer-coated SSP was used. The unstripped cane yield had a strong positive correlation with the stripped cane yield, millable canes, and the number of internodes. Moreover, the commercial cane sugar % resulted in a strong positive correlation with the pol%, whereas the cane sugar recovery % revealed a strong positive correlation with the pol% and commercial cane sugar %.

Optimizing Sugarcane Growth, Yield, and Quality in Different Ecological Zones and Irrigation Sources Amidst Environmental Stressors.

The imbalanced use of fertilizers and irrigation water, particularly supplied from groundwater, has adversely affected crop yield and harvest quality in sugarcane (Saccharum officinarum L.). In this experiment, we evaluated the impact of potassium (K) and micronutrients [viz. Zinc (Zn), Iron (Fe), and Boron (B)] application and irrigation water from two sources, viz. canal, and tube well water on sugarcane growth, yield, and cane quality under field trails. Water samples from Mardan (canal water) and Rahim Yar Khan (tube well water) were analyzed for chemical and nutritional attributes. The results revealed that tube well water's electrical conductivity (EC) was three-fold that of canal water. Based on the EC and total dissolved salts (TDS), 83.33% of the samples were suitable for irrigation, while the sodium adsorption ratio (SAR) indicated only a 4.76% fit and a 35.71% marginal fit compared with canal water. Furthermore, the application of K along with B, Fe, and Zn had led to a significant increase in cane height (12.8%, 9.8%, and 10.6%), cane girth (15.8%, 15.6%, and 11.6%), cane yield (13.7%, 12.3%, and 11.5%), brix contents (14%, 12.2%, and 13%), polarity (15.4%, 1.4%, and 14%), and sugar recovery (7.3%, 5.9%, and 6%) in the tube well irrigation system. For the canal water system, B, Fe, and Zn increased cane height by 15.3%, 13.42%, and 11.6%, cane girth by 13.9%, 9.9%, and 6.5%, cane yield by 42.9%, 43.5%, and 42%, brix content by 10.9%, 7.7%, and 8%, polarity by 33.4%, 28%, and 30%, and sugar recovery by 4.0%, 3.9%, and 2.0%, respectively, compared with sole NPK application. In conclusion, the utilization of tube well water in combination with canal water has shown better results in terms of yield and quality compared with the sole application of canal water. In addition, the combined application of K and B significantly improved sugarcane yields compared with Zn and Fe, even with marginally suitable irrigation water.

Unlocking the potential of glyphosate-resistant bacterial strains in biodegradation and maize growth.

Glyphosate [N-(phosphonomethyl)-glycine] is a non-selective herbicide with a broad spectrum activity that is commonly used to control perennial vegetation in agricultural fields. The widespread utilization of glyphosate in agriculture leads to soil, water, and food crop contamination, resulting in human and environmental health consequences. Therefore, it is imperative to devise techniques for enhancing the degradation of glyphosate in soil. Rhizobacteria play a crucial role in degrading organic contaminants. Limited work has been done on exploring the capabilities of indigenously existing glyphosate-degrading rhizobacteria in Pakistani soils. This research attempts to discover whether native bacteria have the glyphosate-degrading ability for a sustainable solution to glyphosate contamination. Therefore, this study explored the potential of 11 native strains isolated from the soil with repeated glyphosate application history and showed resistance against glyphosate at higher concentrations (200 mg kg-1). Five out of eleven strains outperformed in glyphosate degradation and plant growth promotion. High-pressure liquid chromatography showed that, on average, these five strains degraded 98% glyphosate. In addition, these strains promote maize seed germination index and shoot and root fresh biomass up to 73 and 91%, respectively. Furthermore, inoculation gave an average increase of acid phosphatase (57.97%), alkaline phosphatase (1.76-fold), and dehydrogenase activity (1.75-fold) in glyphosate-contaminated soil. The findings indicated the importance of using indigenous rhizobacteria to degrade glyphosate. Therefore, by maintaining soil health, indigenous soil biodiversity can work effectively for the bioremediation of contaminated soils and sustainable crop production in a world facing food security.