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Abstract Background Cowpea is a high-protein legume that can be grown in many environments. Gamma radiation can modify plant metabolism and growth. An experiment was conducted to determine the effect of different gamma-ray doses on cowpea yield and growth. The results showed that gamma radiation had a significant effect on cowpea yield and growth, with the highest yields and growth rates observed at lower doses. Higher doses had a negative effect on cowpea yield and growth. The study suggests that gamma radiation can be used to improve cowpea yield and growth, but that the optimal dose must be determined for each specific cultivar and environment. Results Results specified that gamma ray significantly affected growth characters and forage yield. Gamma-ray dose of 50 Gy in M1 and M2 generations gave the highest values for all studied characteristics compared to the control and all other irradiation doses, except the crude fiber, followed by 75 and 100 Gy doses for all characters, except number of tillers plant−1 and crude fiber. Seven combinations of SRAP produced 227 loci with an average polymorphism percentage of 85%. The allele frequency of target loci ranged between 0.29 and 0.60, and the PIC was 0.41–0.50 in range, while the GTS% of SRAP combinations ranged from 36 to 63%. The genetic similarities ranged between − 4 and 30%. Ten primers produced 450 loci with a polymorphism percentage of 88%. The allele frequency of target loci ranged between 0.28 and 0.46. The PIC was 0.40–0.50 in range, and the GTS% of IRAP primers ranged from 29 to 71%. The genetic similarities ranged from 8 to 37%. Conclusions SRAP and IRAP analysis revealed more than two clusters of treated plants. In silico analysis showed that some SRAP primers could align with genes in cowpea and related genomes. The SRAP-F12R9 primer is unique to Phaseolus vulgaris, while the SRAP-F13R15 and SRAP-F13em1 primers align with genes on different chromosomes in cowpea cultivar Xiabao 2.

Salinity is a significant abiotic stress that has a profound effect on growth, the content of secondary products, and the genotoxicity of cells. Lime, Citrus aurantifolia, is a popular plant belonging to the family Rutaceae. The interest in cultivating this plant is due to the importance of its volatile oil, which is included in many pharmaceutical industries, but C. aurantifolia plants are affected by the NaCl salinity levels. In the present study, a comet assay test has been applied to evaluate the genotoxic impact of salinity at 0, 50, 100, and 200 mM of NaCl on C. aurantifolia tissue-cultured plants. Furthermore, terpene gene expression was investigated using a semi-quantitative real-time polymerase chain reaction. Results from the two analyses revealed that 200 mM of NaCl stress resulted in high levels of severe damage to the C. aurantifolia plants’ DNA tail 21.8%, tail length 6.56 µm, and tail moment 3.19 Unit. The relative highest expression of RtHK and TAT genes was 2.08, and 1.693, respectively, when plants were exposed to 200 mM of NaCl, whereas pv4CL2RT expressed 1.50 in plants subjected to 100 mM of NaCl. The accumulation of transcripts for the RTMYB was 0.951 when plants were treated with NaCl at 50 mM, and RtGPPS gene was significantly decreased to 0.446 during saline exposure at 100 mM. We conclude that the comet assay test offers an appropriate tool to detect DNA damage as well as RtHK, TAT, and pv4CL2RT genes having post-transcriptional regulation in C. aurantifolia plant cells under salinity stress. Future studies are needed to assess the application of gene expression and comet assay technologies using another set of genes that show vulnerability to different stresses on lime and other plants.

A number of obstacles, including irrigated wastewater and soil contamination, arise in the growth of aromatic and medicinal plants. This study aimed to reduce the effects of contaminated soil and wastewater irrigation on rosemary ( Rosmarinus officinalis L.) plants by using biosynthesized silver nanoparticles (AgNPs) produced by the ginger ( Zingiber officinale ) plant extract. The AgNPs were characterized using Attenuated Total Reflection Fourier Transform Infrared (ATR-FTIR). The experimental design involved three distinct groups of plants: one group was irrigated with regular tap water, another group was rooted in soil contaminated by sewage-wastewater and irrigated with processed wastewater, and the final group consisted of plants grown in wastewater-contaminated soil, irrigated with processed wastewater, and sprayed with 200 mM l ^−1 AgNPs. The study also examined the impact of different treatments on gene expression and secondary metabolite levels in rosemary plants. According to HPLC investigations, nineteen phenol compounds and flavonoids were identified in a methanolic extract of rosemary that was grown in contaminated soil, irrigated with wastewater, and sprayed with AgNPs. Plants treated with wastewater and nanoparticles produced quantities of secondary compounds, including resvertol, vanillic acid, and gallic acid with 1.11, 0.15, and 0.01 mg g ^−1 respectively, which are all regarded as significant antioxidants employed in the pharmaceutical industry. Hexokinase synthase (HK), geranyl diphosphate synthase (GPPS), and linalool synthase (LS) coding genes were found to have highly expressed expressions when plants grown in contaminated soil, wastewater-irrigated plants, and nanoparticle-sprayed plants, respectively, at a 23.2- and 5.54-fold level, where the HK gene was 8.7 times more strongly expressed. Conversely, plants grown in contaminated soil and irrigated with treated wastewater showed downregulation of these genes. Conclusively, using silver nanoparticles significantly reduced the influence of wastewater pollution on secondary metabolites in rosemary plants, which was increased by the gene expression results and was completely consistent with HPLC analysis.

Abstract The release of effluents containing cadmium ions into aquatic ecosystems is hazardous to humans and marine organisms. In the current investigation, biosorption of Cd2+ ions from aqueous solutions by freely suspended and immobilized Turbinaria ornata biomasses was studied. Compared to free cells (94.34%), the maximum Cd2+ removal efficiency reached 98.65% for immobilized cells obtained via Box–Behnken design under optimized conditions comprising algal doses of 5.04 g L−1 and 4.96 g L−1, pH values of 5.06 and 6.84, and initial cadmium concentrations of 25.2 mg L−1 and 26.19 mg L−1, respectively. Langmuir, Freundlich, and Temkin isotherm models were suitably applied, providing the best suit of data for free and immobilized cells, but the Dubinin–Radushkevich model only matched the immobilized algal biomass. The maximum biosorption capacity of Cd2+ ions increased with the immobilized cells (29.6 mg g−1) compared to free cells (23.9 mg g−1). The Cd2+ biosorption data obtained for both biomasses followed pseudo-second-order and Elovich kinetic models. In addition, the biosorption process is controlled by film diffusion followed by intra-particle diffusion. Cd2+ biosorption onto the free and immobilized biomasses was spontaneous, feasible, and endothermic in nature, according to the determined thermodynamic parameters. The algal biomass was further examined via SEM/EDX and FTIR before and after Cd2+ biosorption. SEM/EDX analysis revealed Cd2+ ion binding onto the algal surface. Additionally, FTIR analysis confirmed the presence of numerous functional groups (hydroxyl, carboxyl, amine, phosphate, etc.) participating in Cd2+ biosorption. This study verified that immobilized algal biomasses constitute a cost-effective and favorable biosorbent material for heavy metal removal from ecosystems.

The rose aphid Macrosiphum rosae (L.) is now a globally spread insect species damages rose plants affecting quality and productivity. Botanical insecticides are excellent alternative to synthetic pesticides, as they have minimal environmental persistence and toxicity, and are more compatible with the biocontrol agents than synthetic pesticides. This study aimed to evaluate extracts of five plant species i.e. Citrullus colocynthis, Tagetes erecta, Rosmarinus officinalis, Thymus vulgaris, and Withania somnifera; and entomopathogenic bacteria (EPB), Xenorhabdus budapestensis against M. rosae, as individual and concomitant treatments to determine their compatibility under laboratory conditions. Results indicated that the five plants extracts and EPB applied individually had immense contact or residual toxicity against M. rosae. Methanol extract of T. erecta significantly proved to be more effective as aphicide than ethanol and acetone extracts of five tested plants. Similarly, the results also show a direct, significant relationship between the mortality rates and both EPB cell suspension concentration and exposure time when applied individually. Moreover, three days after treatment, the combination of EPB and each plant extract resulted in a significantly higher M. rosae mortality than the EPB or plant extract alone. We conclude that five plants extracts especially T. erecta had compatible capacity with EPB, thus it could be used in integrated aphid management programs.

The current work is aimed at isolating and identifying new Entomopathogenic bacterium (EPB) strains associated with Steinernema feltiae and assessing the EPB’s biocontrol potential on Aphis punicae and Aphis illinoisensis adults in the laboratory. From S. feltiae, five bacterial isolates were isolated and molecularly characterized. Lysinibacillus xylanilyticus strain TU-2, Lysinibacillus xylanilyticus strain BN-13, Serratia liquefaciens strain TU-6, Stenotrophomonas tumulicola strain T5916-2-1b, and Pseudochrobactrum saccharolyticum strain CCUG are the strains. Pathogenicity tests demonstrated that bacterial cells were more toxic against the two aphid species than bacterial cell-free supernatants. S. tumulicola strain T5916-2-1b cells and filtrate were reported to have the strongest potential to kill A. punicae and A. illinoisensis individuals within 6 h after treatment, with 100% mortality of both insects 24 and 48 h after treatment. Based on the results of the study, it looked like endogenous Steinernema-associated EPB could be used directly as a biocontrol agent for A. punicae and A. illinoisensis.

Fusarium oxysporum is one of the most harmful soil-borne pathogens that cause root rot, damping-off, and wilt disease in many plant species. Management of Fusarium oxysporum diseases is often by using many harmful and expensive chemical fungicides which have many harmful effects on the environment and human health. The current study was conducted to identify the chemical constituents of black cumin seeds’ methanolic extract and investigate the ability of the major constituents to inhibit the Fusarium oxysporum trypsin-like serine protease, which play an important role in F. oxysporun pathogenicity. The HPLC-MS analysis of black cumin seeds’ methanolic extract revealed the presence of seven major compounds: amentoflavone, Procyanidin C2, Quercetin3-O-sophoroside-7-O-rhamnoside, 5,7-Dihydroxy-3,4-dimethoxyflavone, Borapetoside A, tetrahydroxy-urs-12-en-28-O-[b-D-glucopyranosyl (1-2)-b-D-glucopyranosyl] ester, and kudzusapongenol A-hexA-pen. The results of molecular docking between these compounds and the active site of Fusarium oxysporium trypsin showed that only four compounds were able to bind to the active site of F. oxysporum trypsin. Amentoflavone, 5,7-Dihydroxy-3,4-dimethoxyflavone, and Quercetin3-O-sophoroside-7-O-rhamnoside have the highest binding energy, −6.4, −6.5, and −6.5 Kcal/mol, respectively. In addition, the results clarify that 5,7-Dihydroxy-3,4-dimethoxyflavone was the only compound to form a hydrogen bond with Asp189 (the residue responsible for substrate specificity). The results of the study strongly indicate that flavonoids of black cumin seeds’ methanolic extract could be used as effective inhibitors for the F. oxysporum trypsin-like serine protease.

Silybum marianum L. commonly known as milk thistle is a medicinally potent plant with a multitude of pharmacological applications. The present investigations demonstrated the effects of Zinc Oxide nanoparticles (ZnO NPs) on callus growth and biosynthesis of silymarin in milk thistle under various light conditions. The callus cultures developed on Murashige and Skoog (MS) basal media containing ZnO NPs (0.15 mg/L), under the dark condition maintained for two weeks, followed by transference into normal light produced the maximum callus fresh weight (2294 mg/L FW). Further, the metabolite profiling revealed that ZnO NPs significantly augmented the production of silymarin and upregulated the antioxidant system in the callus cultures. Maximum TPC (total phenolic content: 37 ± 0.20 mg/g DW), TFC (total flavonoid content: 8.9 ± 0.023), DPPH antioxidant activity (91.5 ± 1.75%), Superoxide dismutase activity (SOD: 4.1 ± 0.045 nM/min/mg FW) and the highest silymarin content (14.6 ± 0.023 mg/g DW) were recorded in the callus cultures developed on MS media supplemented with solitary ZnO NPs (0.15 mg/L). While the callus culture evolved in presence of only PGRs (2,4 D and BA: 2 mg/L, each) accumulated the lesser fresh weight (562 mg/L FW). A higher concentration of ZnO NPs (0.15 mg/L) enhanced the secondary metabolite accumulation and silymarin content in the callus of Silybum marianum. This is the first standardized protocol to be applied on the industrial level for the production of silymarin.

Soybean is a legume crop enriched with proteins and oil. It is frequently exposed to anthropogenic and natural flooding that limits its growth and yield. Current study applied gel-free proteomic techniques to unravel soybean response mechanism to flooding stress. Two-days-old soybeans were flooded for 4 days continuously and root samples were collected at days 2 to 6 for proteomic and enzymatic analyses. Age-matched untreated soybeans were collected as control. After protein extraction, purification and tryptic digestion, the peptides were analyzed on nano-liquid chromatography-mass spectrometry. A total of 539 and 472 proteins with matched peptides 2 or more were identified in control and flooded seedlings, respectively. Among these 364 proteins were commonly identified in both control and flooded soybeans. Fourty-two protein's abundances were changed 4-fold after 2-days of flooding stress as compared to starting point. The cluster analysis showed that highly increased proteins included cupin family proteins, enolase, pectin methylesterase inhibitor, glyoxalase II, alcohol dehydrogenase and aldolase. The enzyme assay of enolase and pectin methylesterase inhibitor confirmed protein abundance changes. These findings suggest that soybean adopts the less energy consuming strategies and brings biochemical and structural changes in the cell wall to effectively respond to flooding stress and for the survival.

A field experiment was conducted during the Rabi season 2017–2018 (October–March) at the University of Agriculture, Peshawar research farm to examine the influence of different nitrogen (N) and phosphorus (P) levels on two different oat varieties: Australian and Ukrainian. The treatments included control and three levels of nitrogen and phosphorus at 30, 60, and 90 kg ha−1. The treatments were arranged in randomized complete block design (RCBD) and replicated three times. The findings showed that the oat varieties were significantly different from one another in yield and yield parameters. The Australian variety recorded higher emergence (49 plants m−2), days to emergence (15 days), days to flowering (122 days), days to maturity (145 days), plant height (142.7 cm), number of leaves (6.03 leaves plant−1), number of tillers (92.2 tillers m−1), biological yield (8,179.2 kg ha−1), and grain yield (3,725.6 kg ha−1) than the Ukrainian variety. Similarly, different N and P levels, the maximum days to emergence, days to flowering, and days to maturity were recorded in a control plot. The application of 105 kg N + 90 kg P ha−1 was statistically similar to the application of 105 kg N + 60 kg P ha−1. Maximum emergence (60 plants m−2), number of leaves (7.0 leaves plant−1), plant height (118.6 cm), number of tillers m−1 (102.6), biological yield (9,687.5 kg ha−1), and grain yield (4,416.7 kg ha−1) were determined in Australian variety. Based on the findings of this study, the Australian variety performed better in terms of yield and yield components and the application of N and P fertilizers at the rate of 105 kg N + 60 kg P ha−1 produced the best results in both oat varieties.

The principal intent of the investigation was to examine the influence of joint application of methyl jasmonate (MeJA, 10 μM) and a nitric oxide–donor sodium nitroprusside (SNP, 100 μM) to wheat plants grown under cadmium (Cd as CdCl2, 100 μM) stress. Cd stress suppressed plant growth, chlorophylls (Chl), and PSII maximum efficiency (Fv/Fm), but it elevated leaf and root Cd, and contents of leaf proline, phytochelatins, malondialdehyde, and hydrogen peroxide, as well as the activity of lipoxygenase. MeJA and SNP applied jointly or singly improved the concentrations of key antioxidant biomolecules, e.g., reduced glutathione and ascorbic acid and the activities of the key oxidative defense system enzymes such as catalase, superoxide dismutase, dehydroascorbate reductase, glutathione S-transferase, and glutathione reductase. Exogenously applied MeJA and SNP jointly or singly also improved nitrogen metabolism by activating the activities of glutamine synthetase, glutamate synthase, and nitrate and nitrite reductases. Compared with individual application of MeJA or SNP, the combined application of both showed better effect in terms of improving plant growth and key metabolic processes and reducing tissue Cd content, suggesting a putative interactive role of both compounds in alleviating Cd toxicity in wheat plants.Main findingsThe main findings are that exogenous application of methyl jasmonate and nitric oxide–donor sodium nitroprusside alleviated the cadmium (Cd)–induced adverse effects on growth of wheat plants grown under Cd by modulating key physiological processes and up-regulating enzymatic antioxidants and the ascorbic acid–glutathione cycle–related enzymes.

Chemotherapy is an aggressive form of chemical drug therapy aiming to destroy cancer cells. Adjuvant therapy may reduce hazards of chemotherapy and help in destroying these cells when obtained from natural products, such as medical plants. In this study, the potential therapeutic effect of Rosa damascena callus crude extract produced in vitamin-enhanced media is investigated on colorectal cancer cell line Caco-2. Two elicitors, i.e., L-ascorbic acid and citric acid at a concentration of 0.5 g/L were added to the callus induction medium. Callus extraction and the GC–MS analysis of methanolic crude extracts were also determined. Cytotoxicity, clonogenicity, proliferation and migration of Caco-2 colorectal cancer cells were investigated using MTT cytotoxicity, colony-forming, Ki-67 flow cytometry proliferation and Migration Scratch assays, respectively. Our results indicated that L-ascorbic acid treatment enhanced callus growth parameters and improved secondary metabolite contents. It showed the least IC50 value of 137 ug/mL compared to 237 ug/mL and 180 ug/mL in the citric acid-treated and control group. We can conclude that R. damascena callus elicited by L-ascorbic acid improved growth and secondary metabolite contents as well as having an efficient antiproliferative, anti-clonogenic and anti-migratory effect on Caco-2 cancer cells, thus, can be used as an adjuvant anti-cancer therapy.

Aphids are serious pests of a wide range of agricultural crops, including pomegranates and grapevines. In addition, due to the negative environmental impacts of chemical insecticides, these pests are developing important resistance against aphicides. Therefore, one alternative method to control aphids is the use of essential oils (EO). The present study aimed to evaluate the insecticidal activity of Citrus aurantium and C. reticulata peel EO at different concentrations and with different exposure periods to pomegranate and grapevine aphids, Aphis punicae and A. illinoisensis via the topical application method under laboratory conditions. The results reveal that C. aurantium L. EO had greater toxicity against pomegranate and grapevine aphids, with LC50 of 0.37 and 0.82 μL/mL, respectively, at 48 h after application. The highest repellence effect was estimated for C. aurantium EO, at 2.5 μL/cm2, on A. punicae, with a value of 100% after an exposure time of 3 h, in contrast to the 88% repellence estimated for A. illinoisensis. The GC-MS investigation of both essential oils identified limonene, 3-carene, pinene, and p-cymene as active substances that could be attributed to the effects observed. Overall, our results offer a potential tool to control the two aphid species and could help in the development of integrated insect management in pomegranate and grapevine fields.

The rice weevil, Sitophilus oryzae (L.), is the most destructive insect pest of stored cereals worldwide. The current study was conducted to determine the lethal, reproductive, and histological effects of gamma irradiation on S. oryzae adults. In addition, the impact on germination, chlorophyll, and proline content in wheat seedlings from treated grains was determined. Wheat grains were infested with rice weevil adults and then irradiated by gamma rays. Gamma radiation was applied at a dosage of 0.10, 0.25, 0.50, and 1.00 kGy. Mortality percentage and LD50 were recorded after 48, 72, 96, and 120 h of treatment. The dosage of 1.00 kGy caused 100% mortality after 96 h of irradiation. The required dosage of gamma radiation to kill 50% (LD50) of adults after 48 h was 1.51 kGy. All tested doses caused complete sterility to 24 h old adults. A histological alteration was noticed at a dosage of 1.00 kGy, which showed cytoplasmic vacuolization, tissues exhibiting signs of putrefaction, and necrosis of cells; furthermore, gamma irradiation affected chlorophyll a and b. The highest amounts were detected in wheat seedlings from grains irradiated at 0.10 kGy. There was a significant increase in plant proline content at the higher doses (0.50 and 1.00 kGy) compared with seedlings from nonirradiated grains. It could be concluded that gamma radiation can be used as an eco-friendly trend to control stored-product pests without any residual effects.

Entomopathogenic bacterium, Xenorhabdus has a mutualistic relationship with entomopathogenic nematode of the genus Steinernema and produces several bio-agent compounds with antimicrobial and nematicidal activities. Root-knot nematodes are considered one of the most important pests facing the cultivation of grapevine worldwide. A micro-plot field trial was conducted in naturally infested soil with Meloidogyne incognita to evaluate the potential of two strains of entomopathogenic bacteria namely Xenorhabdus budapestensis DSM 16342 (EMA) and X. szentirmaii DSM 16338 (EMC) applied separately or integrated with neem cake and/or furadan at half of recommended dose on nematode development and growth improvement of Taify grapevine. Data of nematode populations, number of galls and egg-masses, eggs/g root, plant lengths and weights and number of leaves were recorded four months after application. Results appeared significant differences between treatments and control. The triple application was more effective than dual and single applications in reducing nematode infestation and improving plant growth. Combined application of EMC or EMA with furadan or neem cake increased the efficacy (64.6-68.6%) and improved plant fresh weight (27.4-69.5%). Conclusively, utilization of such bacterial filtrates with either neem cake and/or nematicide could gain a successful approach in integrated nematode management programs.

Many phytochemicals can affect the growth and development of plants and insects which can be used as biological control agents. In this study, different concentrations of crude, hexane, chloroform, butanol, and aqueous extracts of Euphorbia nivulia Buch.-Ham., an endemic plant of the Cholistan desert in South Punjab of Pakistan, were analysed for their chemical constituents. Their various concentrations were also tested for their phytotoxic and insecticidal potential against duckweed, Lemna minor L., and the dusky cotton bug, Oxycarenus hyalinipennis Costa. various polyphenols, i.e., quercetin, gallic acid, caffeic acid, syringic acid, coumaric acid, ferulic acid, and cinnamic acid were detected in different concentrations with different solvents during the phytochemical screening of E. nivulia. In the phytotoxicity test, except for 100 μg/mL of the butanol extract gave 4.5% growth regulation, no phytotoxic lethality could be found at 10 and 100 μg/mL of all the extracts. The highest concentration, 1000 μg/mL, of the chloroform, crude, and butanol extracts showed 100, 63.1, and 27.1% of growth inhibition in duckweed, respectively. In the insecticidal bioassay, the highest O. hyalinipennis mortalities (87 and 75%) were recorded at 15% concentration of the chloroform and butanol extracts of E. nivulia. In contrast, the lower concentrations of the E. nivulia extracts caused the lower mortalities. Altogether, these findings revealed that E. nivulia chloroform extracts showed significant phytotoxicity while all the extracts showed insecticidal potential. This potential can be, further, refined to be developed for bio-control agents.

Abiotic stress, especially a lack of water, can significantly reduce crop yields. In this study, we evaluated the physiological and biochemical effects of potassium sulfate (K2SO4) fertilizer and varied irrigation regimes on the economically significant oilseed crop, Brassica juncea L, under open field conditions. Two cultivars (RH-725 and RH-749) of B. juncea were used in a randomized complete block design experiment with three replicates. Irrigation regimes consisted of a control (double irrigation: once at the 50% flowering and another at 50% fruiting stages), early irrigation (at 50% flowering only), late irrigation (at 50% fruiting only) and stress (no irrigation). The K2SO4 applications were: control (K0, no fertilization); K1, 10 kg ha-1; and K2, 20 kg ha-1. We measured growth via fresh and dry plant weight, plant height, root length, and leaf area. All the growth parameters were higher in RH-749. The physiological attributes, including the membrane stability index and relative water content, were higher at the 50% flowering stage in RH-749. The amount of antioxidant enzymes (catalase (CAT), guaiacol peroxidase (POX), ascorbate peroxidase (APX), and superoxide dismutase (SOD)) was enhanced when both plants were fertilized during water stress. All of these enzymes had higher activity in RH-749. The total chlorophyll content and photosynthesis rate were considerably higher in RH-749, which leaked fewer electrolytes and maintained a less destructive osmotic potential under limited water conditions. The results indicated that it is water-stress tolerant when given a high concentration of K2SO4, which alleviated the adverse effects of water stress on growth and physiology.

Virachola livia (Lepidoptera: Lycaenidae) and Ectomyelois ceratoniae (Lepidoptera: Pyralidae) are the key pests of pomegranates in Saudi Arabia that are managed mainly using broad-spectrum pesticides. Interactions between the entomopathogenic nematodes (EPNs) Steinernematids, and Heterorhabditids, and their entomopathogenic bacterial symbionts (EPBs) have long been considered monoxenic 2-partner associations responsible for killing insects and, therefore, are widely used in insect pest biocontrol. However, there are limited reports identifying such organisms in Taif, Saudi Arabia. The current study aimed to identify the EPNs and their associated bacteria isolated from Taif, Saudi Arabia, and evaluate their biocontrol potential on third instar larvae of V. livia and E. ceratoniae under laboratory conditions. A total of 35 EPN isolates belonging to Steinernema (20) and Heterorhabditis (15) were recovered from 320 soil samples. Twenty-six isolates of symbiotic or associated bacteria were isolated from EPNs and molecularly identified as Xenorhabdus (6 isolates), Photorhabdus (4 isolates), Pseudomonas (7), or Stenotrophomonas (9). A pathogenicity assay revealed that Steinernema spp. were more virulent than Heterorhabditis spp. against the two pomegranate insects, with LC50 values of 18.5 and 13.6 infective juveniles (IJs)/larva of V. livia for Steinernema spp. and 52 and 32.4 IJs/larva of V. livia for Heterorhabditis spp. at 48 and 72 h post-treatment, respectively. Moreover, LC50 values of 9 and 6.6 IJs/larva (Steinernema spp.) and 34.4 and 26.6 IJs/larva (Heterorhabditis spp.) were recorded for E. ceratoniae larvae at 48 and 72 h post-treatment. In addition, the EPB Stenotrophomonas maltophilia CQ1, isolated from Steinernema spp., surpassed Pseudomonas mosselii SJ10, associated with Heterorhabditis spp., in their ability to kill V. livia or E. ceratoniae larvae within 6 h post-application, resulting in 100% mortality in both insects after 24 and 48 h of exposure. We conclude that either application of EPNs’ IJs or their associated EPBs could serve as potential biocontrol agents for V. livia and E. ceratoniae.

Ectomyelois ceratoniae (Lepidoptera: Pyralidae) is the primary pest of pomegranates in Saudi Arabia and is mostly controlled using broad-spectrum pesticides. Providing environmentally sound choices to limit reliance on chemical management is a major challenge in the control of E. ceratoniae and, as a consequence, in the protection of pomegranate crops from its invasion. Entomopathogenic bacteria (EPB) symbiotically associated with entomopathogenic nematodes (EPNs) are well-known biocontrol agents of soil-dwelling or aerial pests. The bacterium symbiont (EPB) is the real insect-killing biocontrol agent, while the nematode (EPN) serves as a vector. We wondered whether the EPB vector, which is extremely vulnerable to adverse environmental conditions, like drought, high temperatures, and repellent soil microorganisms, could be omitted. We intended to evaluate the biocontrol potential of directly applied EPB cells and cell-free culture media (CFCM) on the larval instar E. ceratoniae. Xenorhabdus budapestensis DSM 16342 (EMA), X. szentirmaii DSM 16338 (EMC), and Photorhabdus luminescens ssp. laumondi (TT01) strains were used. After three days of exposure, the cells of EMA, EMC, and TT01 strains resulted in 100%, 88%, and 79.3% larval mortality rates, respectively. The applied EMA CFCM resulted in 53.7% larval mortality, indicating the presences of (at least) one extremely strong component produced by EMA. We concluded that the direct application of either the EPB cells or the CFCM must be a prospective alternative biocontrol of E. ceratoniae, especially to protect the important fruit (pomegranate, Punica granatum) cultivars. Especially, newly identified local EPB isolates could be applied as bio-pesticides for integrated management practices or organic pomegranate production.

The large genetic evolution due to the sexual reproduction-mediated gene assortments and propensities has made Venturia inaequalis (causing apple scab) unique with respect to its management strategies. The resistance in apple germplasm against the scab, being controlled for by more than fifteen genes, has limited gene alteration-based investigations. Therefore, a biological approach of bacterial endophyte community dynamics was envisioned across the apple germplasm in context to the fungistatic behavior against V. inaequalis. A total of 155 colonies of bacterial endophytes were isolated from various plant parts of the apple, comprising 19 varieties, and after screening for antifungal behavior followed by morphological, ARDRA, and sequence analysis, a total of 71 isolates were selected for this study. The alpha diversity indices were seen to fluctuate greatly among the isolation samples in context to microflora with antifungal behavior. As all the isolates were screened for the presence of various metabolites and some relevant genes that directly or indirectly influence the fungistatic behavior of the isolated microflora, a huge variation among the isolated microflora was observed. The outstanding isolates showing highest percentage growth inhibition of V. inaequalis were exploited to raise a bio-formulation, which was tested against the scab prevalence in eight apple varieties under controlled growth conditions. The formulation at all the concentrations caused considerable reductions in both the disease severity and disease incidence in all the tested apple varieties. Red Delicious being most important cultivar of the northwestern Himalayas was further investigated for its biochemical behavior in formulation and the investigation revealed different levels of enzyme production, chlorophyll, and sugars against the non-inoculated control.

This study was conducted on tomato (Solanum lycopersicum cv. K-21) to investigate the bioprotective nature of Pseudomonas fluorescens and its interactive effects with Meloidogyne incognita in terms of growth biomarkers, changes in biochemical attributes and modulation in antioxidant enzymes of the tomato plant. In this study, we grew tomato plants with M. incognita and P. fluorescens in separate pots, simultaneously and sequentially (15 days prior or post) after 15 days of seed sowing. The sequential inoculation of Mi15→Pf maximally increased the root-knot index and decreased the nematode population. It was also noted that inoculation suppressed the plant growth biomarkers in comparison to control. However, maximum suppression in nematode reproduction and increment in growth and physiological attributes were observed when P. fluorescens was applied 15 days prior to the nematode (Pf15→Mi) as compared to control. All the treatments showed an increase in antioxidant enzymes. Expression of phenol content and defensive enzymes such as peroxidase (POX) and superoxide dismutase (SOD) increased, in contrast to a significant reduction in malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents when compared with the untreated inoculated plants. However, the highest levels of POX and SOD, and a lowest of phenol, MDA and H2O2 were displayed in the treatment Pf15→Mi, followed by Mi+Pf and Mi15→Pf.

The presence of inorganic pollutants such as Cadmium(II) and Chromium(VI) could destroy our environment and ecosystem. To overcome this problem, much attention was directed to microbial technology, whereas some microorganisms could resist the toxic effects and decrease pollutants concentration while the microbial viability is sustained. Therefore, we built up a complementary strategy to study the biofilm formation of isolated strains under the stress of heavy metals. As target resistive organisms, Rhizobium-MAP7 and Rhodotorula ALT72 were identified. However, Pontoea agglumerans strains were exploited as the susceptible organism to the heavy metal exposure. Among the methods of sensing and analysis, bioelectrochemical measurements showed the most effective tools to study the susceptibility and resistivity to the heavy metals. The tested Rhizobium strain showed higher ability of removal of heavy metals and more resistive to metals ions since its cell viability was not strongly inhibited by the toxic metal ions over various concentrations. On the other hand, electrochemically active biofilm exhibited higher bioelectrochemical signals in presence of heavy metals ions. So by using the two strains, especially Rhizobium-MAP7, the detection and removal of heavy metals Cr(VI) and Cd(II) is highly supported and recommended.

Resistance of bacteria to multiple antibiotics is a significant health problem; hence, to continually respond to this challenge, different antibacterial agents must be constantly discovered. In this work, fluoroaryl-2,2′-bichalcophene derivatives were chemically synthesized and their biological activities were evaluated against Staphylococcus aureus (S. aureus). The impact of the investigated bichalcophene derivatives was studied on the ultrastructural level via scanning electron microscopy (SEM), molecular level via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method and on the biofilm inhibition via the electrochemical biosensors. Arylbichalcophenes’ antibacterial activity against S. aureus was affected by the presence and location of fluorine atoms. The fluorobithiophene derivative MA-1156 displayed the best minimum inhibitory concentration (MIC) value of 16 µM among the tested fluoroarylbichalcophenes. Over a period of seven days, S. aureus did not develop any resistance against the tested fluoroarylbichalcophenes at higher concentrations. The impact of fluoroarylbichalcophenes was strong on S. aureus protein pattern showing high degrees of polymorphism. SEM micrographs of S. aureus cells treated with fluoroarylbichalcophenes displayed smaller cell-sizes, fewer numbers, arranged in a linear form and some of them were damaged when compared to the untreated cells. The bioelectrochemical measurements demonstrated the strong sensitivity of S. aureus cells to the tested fluoroarylbichalcophenes and an antibiofilm agent. Eventually, these fluoroarylbichalcophene compounds especially the MA-1156 could be recommended as effective antibacterial agents.