Evaluating the genotoxicity of salinity stress and secondary products gene manipulation in lime, Citrus aurantifolia, plants.

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.

Influence of Effective Microorganisms on Some Biological and Biochemical Aspects of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae).

The cotton leafworm, Spodoptera littoralis (Bosid.), is a major pest in African and Asian nations that attacks a wide variety of host plants. This study was conducted to assess the effectiveness of effective microorganisms (EMs) on the biological and physiological features of S. littoralis larvae. Five concentrations (100, 200, 300, 400, and 500 ppm) of EMs were tested. Antifeedant activity, food consumption index, the efficiency of converting digested food, the efficiency of converting ingested food, relative growth rate, and approximate body tissue of the fourth larval instar of S. littoralis were determined. Moreover, carbohydrate enzyme activities (amylase, trehalose, and invertase), total protein, and total lipids of S. littoralis larvae were measured to elucidate the mode of action of the tested agent in the S. littoralis's larval stage. The EMs at 500 ppm had a substantial impact on antifeedant activity, nutritional indices, egg deposit reduction, and hatchability in S. littoralis during the five days. All concentrations interrupted S. littoralis's life cycle and developmental phases. Furthermore, all concentrations were quite useful in lengthening the developmental stages of S. littoralis. In addition, Ems affected the biochemical activities of larvae, leading to disturbances in carbohydrate, lipid, and protein levels. From this study, EMs can be used as a bioinsecticide alternative to traditional insecticides against S. littoralis and may be compatible with integrated pest management approaches.

Identification and environment-friendly biocontrol potential of five different bacteria against Aphis punicae and Aphis illinoisensis (Hemiptera: Aphididae).

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.

Evaluation of the Anti-Cancer Potential of Rosa damascena Mill. Callus Extracts against the Human Colorectal Adenocarcinoma Cell Line.

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.

Gamma Radiation: An Eco-Friendly Control Method for the Rice Weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae).

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.

Standardization of in vitro micropropagation of Winter Jasmine (Jasminum nudiflorum) using nodal explants.

Present investigation was carried out to arrive at an effective micropropagation protocol for Winter Jasmine (Jasminum nudiflorum) using nodal segments from actively growing plants as explants. Explants were collected from current season shoots during April-May just after the initiation of new flush. Combined sterilization treatment of explants with 1.0% NaOCl2 for 10 min followed by 70% ethanol for 10 s recorded highest culture survival (63.88%) and optimum culture asepsis (63.88%) followed by the treatment containing 0.1% HgCl2 for 10 min followed by 70% ethanol for 10 s with culture survival (61.11%) and culture asepsis (69.44%). Highest culture establishment (80.55%) and minimum days to bud sprouting (7.62 days) was recorded with Benzyl adenine + Kinetin (3.0 + 1.0 mgL-1) but maximum length (4.33 cm) and leaf number (7.78) of established micro shoots was recorded with Benzyl adenine + Kinetin (1.0 + 0.5 mgL-1). Maximum proliferated shoots (2.41) and an optimum proliferation percentage (77.78 %) was recorded with Benzyl adenine + Kinetin (3.0 + 0.5 mgL-1). Minimum size of proliferated shoots (2.02 cm) was recorded with Benzyl adenine + Kinetin (3.0 + 1.0 mgL-1) followed by 2.25 cm recorded with Benzyl adenine + Kinetin (3.0 + 0.5 mgL-1). Highest rooting (63.93%), primary root number/microshoot (4.74) and longest primary roots (34.67 mm) were recorded with IBA (2.0 mgL-1). IBA yielded better results than NAA in terms of higher rooting percentage and root number. However, days to root initiation were found minimum (22.00) with 2.0 mgL-1 of NAA. Highest ex vitro survival of rooted microshoots (89.67%) was recorded with IBA (2.0 mgL-1).

Assessment of variability among morphological and molecular characters in wild populations of mint [Mentha longifolia (L.) L.] germplasm.

Mentha longifolia is an important medicinal and aromatic perennial herb that exhibits wide distribution range from sub-tropical to temperate regions. In the present study, agro-morphological traits and genetic differences in 19 different populations of M. longifolia were studied to evaluate the level and extent of its diversity. Analysis of variance (ANOVA) showed that the different phenotypic characters show considerable differences among various populations and was significant at p < 0.05. Molecular diversity analysis performed by using arbitrary amplified eleven ISSR primers generated a total of 121 amplicons that range within the size of 200-2500 base pairs (bp). Each primer on average generated 11 amplicons with percentage polymorphism being 100. The analysis of molecular variance (AMOVA) showed more (64%) among population genetic diversity and less (36%) within the populations. Greater genetic differentiation (Gst = 0.6852) among these populations occurs due to low gene flow (Nm = 0.2297) and greater habitat variability. Geographic and genetic distances were positively correlated according to Mantel's test. In order to remove any kind of biases, we used R software to perform cluster and redundancy analysis to analyse the extent of relatedness among studied populations. In terms of morphological and molecular aspects, the populations were grouped into four and five clusters respectively based on hierarchical clustering method. The results demonstrated that M. longifolia displays a great degree of morphological and genetic variation and can be utilized in breeding, genetic improvement, and gene bank conservation programmes in future.

Proteomic insight into soybean response to flooding stress reveals changes in energy metabolism and cell wall modifications.

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.

Process optimization and modeling of Cd2+ biosorption onto the free and immobilized Turbinaria ornata using Box-Behnken experimental design.

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.

Application of melatonin and PGPR alleviates thiamethoxam induced toxicity by regulating the TCA cycle in Brassica juncea L.

Thiamethoxam, a broad spectrum, neonicotinoid insecticide, is used on various crops including Brassica juncea L. to protect from intruding insects such as leaf-hoppers, aphids, thrips and white-flies. Exposure to thiamethoxam causes acute malady such as tumour development, cell apoptosis, liver damage and neurotoxicity. Melatonin is entailed in umpteen developmental processes of plants, including stress responses. The pleiotropic effects of melatonin in modulating plant growth validate it's imperative contribution as multi-regulatory substance. Exiguous information is known about the role of Pseudomonas putida in improving plant growth under thiamethoxam stress. Taking these aspects into consideration the contemporary study investigates the role of melatonin and Pseudomonas putida strain MTCC 3315 in alleviating the thiamethoxam induced toxicity in B. juncea plant. Fourier Transform Infrared Spectroscopy (FTIR) analysis uncloaked that thiamethoxam induced stress primarily affects the protein content of plant as compared to lipids, carbohydrates and cell wall components. Organic acid profiling of the treated samples carried-out by High-Performance Liquid Chromatography (HPLC), reported an upregulation in the level of organic acids, malic acid (110%), citric acid (170%), succinic acid (81%), fumaric acid (40%) and ascorbic acid (55%) in thiamethoxam treated plants compared to the investigational untreated plants. The melatonin treated seedlings grown under thiamethoxam stress, exhibit increased level of malic acid, citric acid, succinic acid, fumaric acid and ascorbic acid by 81%, 0.94%, 11%, 21% and 6% respectively. Further, thiamethoxam stressed plants inoculated with Pseudomonas putida showed stupendous up-regulation by 161% (malic acid), by 14% (citric acid), by 33% (succinic acid), by 30% (fumaric acid), by 100% (oxalic acid) respectively. Lastly, the combinatorial application of melatonin and Pseudomonas putida resulted in prodigious upsurge of malic acid by 165%, succinic acid by 69%, fumaric acid by 42% respectively in contrast to distinct melatonin and Pseudomonas putida treatments. The accumulation of organic acids ascertains the defence against thiamethoxam stress and corresponds to meet the energy generation requirement to skirmish thiamethoxam mediated abiotic stress in Brassica juncea plant.

Isolation, Identification, and Biocontrol Potential of Entomopathogenic Nematodes and Associated Bacteria against Virachola livia (Lepidoptera: Lycaenidae) and Ectomyelois ceratoniae (Lepidoptera: Pyralidae).

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.

Identification, characterization and optimization of phosphate solubilizing rhizobacteria (PSRB) from rice rhizosphere.

Two billion people worldwide take rice (Oryza sativa L.) as a staple food. Phosphorus (P) and Nitrogen (N) are the major requirements of rice; although these are available in limited concentrations within rice growing regions. Among different types of Plant growth-promoting rhizobacteria (PGPR), Phosphate solubilizing rhizobacteria (PSRB) constitute an important class. These are known for plant growth promotion by enhancing P and N uptake. PSRB are nowadays used as biofertilizers to restore the soil health. Under the present investigation identification, characterization and optimization of phosphate solubilizing activity of these microbes at different pH, temperature and salt concentrations was carried out. Thirty-seven isolates were recovered from different regions of rice rhizosphere on Pikovskaya (PVK) agar among which 15 isolates were recovered from R.S. Pura, 12 isolates from Bishnah and 10 isolates were recovered from Akhnoor sector of Jammu, India. A prominent halo zone of clearance was developed around the colonies of 12 different isolates, indicating phosphate solubilization activity. Four distinct isolates were amplified, cloned and sequenced for taxonomic identification using 16S primers. The results indicated that PS 1, PS 2, PS 3, PS 4 were related to Pseudomonas aeruginosa, Bacillus subtilis strain 1, B. subtilis strain 2, B. subtilis strain 3, respectively. These strains when grown at a wide range of ecological factors showed maximum growth at pH between 6.8 and 8.8, temperature between 28 °C and 37 °C and salinity between 1% and 2%. Screening for phosphate solubilization activity revealed that the halo zone diameter formed by these isolates extended from 2.1 to 3.2 mm. The phosphate solubilizing efficiency (SE) ranged from 35.4 to 50.9 with highest value of 50.9 by PS4 and maximum P solubilization of 10.22 µg/ml was recorded by PS4 at 7th day. Phosphate solubilization activity of these identified PSRB strains can be utilized and explored in the rice growing belts of Jammu region which are deficient in phosphorus. MIC value for zinc sulphate heptahydrate in 12 isolates varied from 1 mg/ml to 6 mg/ml. Phosphate solubilization activity and MIC of these identified PSRB strains can be utilized and explored in the rice growing belts of Jammu region which are deficient in phosphorus.

Phosphate solubilizing bacteria enhanced growth, oil yield, antioxidant properties and biodiesel quality of Kasumbha.

Biodiesel is considered as a potential alternative energy source, but problem exists with the quantity and quality of feedstock used for it. To improve the feedstock quality of biodiesel, a field experiment was conducted under natural conditions. Cultivar Thori of kasumbha was used in the experiment. Commercialized biofertilizers were applied at the rate of 20 kg per acre and chemical fertilizer (diammonium phosphate) was applied as half dose (15 kg/ha). Results indicated that number of leaf plant-1, leaf area, number of seeds capitulum-1 was significantly increased by biofertilizer treatment alone (BF) and combine treatment of biofertilizer and chemical fertilizer (BFCF). Agronomic traits such as plant height, no. of branches of a plant, no. of capitulum/plant was improved significantly by BF treatment over the control. Maximum 1000 seed weight (41%) and seed yield (23%) were recorded in half dose of chemical fertilizers treatment (CFH). Seed oil content and seed phenolics were significantly improved by BF and CF treatments while maximum biodiesel yield was recorded by BF treatment. Maximum oleic acid was recorded by BF treatment while other fatty acids being maximum in control except linoleic acid in BFCF treatment. Results for specific gravity were non-significant while acid value and free fatty acid contents were substantially reduced by BF treatment as compared to other treatments. Maximum value of iodine number was recorded in BFCF treatment while tocopherol contents were improved by BF treatment. It is inferred that biofertilizer treatment alone perform better as compared to other treatments and 50% chemical fertilizer can be replaced using biofertilizer which is a good approach for sustainable environmental-friendly agriculture.

Drought-tolerant Pseudomonas sp. showed differential expression of stress-responsive genes and induced drought tolerance in Arabidopsis thaliana.

The growth and persistence of rhizobacteria in soils are highly impacted by moisture stress. In this study, we report the first transcript analysis of four Pseudomonas strains (PS1, PS2, PS3, and PS4) isolated from the root-soil interface of rice and maize associated with different moisture levels during water deprivation. Filtered Pseudomonas sp. cells incubated at low (RH10%) and high (RH85%) relative humidity showed decreased survival of all Pseudomonas sp. at RH10% when compared with RH85%. RT-PCR showed differential expression of treS (trehalose synthase), rpoS (sigma factor), mucA (alginate regulatory gene), and fliM (flagellar motor switch protein gene) in response to exposure to RH10%. However, molecular fingerprinting and nutrient assimilation profile of Pseudomonas strains demonstrated genetic and physiological variation between the four strains irrespective of water regime and host. In vitro testing of these strains showed ACC deaminase activity and gibberellic acid, abscisic acid, indole acetic acid, and exopolysaccharide production. We determined that 50 µl of 1.2 × 103 CFU ml-1 of these Pseudomonas strains was enough to protect Arabidopsis plants against drought stress in a pot experiment. Inoculated plants increased their root colonization ability and biomass; however, PS2 showed higher survival (95%), relative water content (59%), chlorophyll (30%), glycine betaine (38%), proline (23%), and reduced MDA (43%) in shoots than irrigated control under induced water deprivation. It can be concluded that all Pseudomonas strains were effective in mitigating drought stress, however, PS2 appears to impart more resistance to drought than the other strains by upregulating key defense mechanisms.

Effects of benzo(a)pyrene on meiobenthic assemblage and biochemical biomarkers in an Oncholaimus campylocercoides (Nematoda) microcosm.

A microcosm experiment was carried out to determine how benzo(a)pyrene (BaP) may affect marine meiofauna community, with a main emphasis on nematode structure and functional traits. Three increasing concentrations of BaP (i.e. 100, 200 and 300 ng/l, respectively) were used for 30 days. The results revealed a gradual decrease in the abundance of all meiobenthic groups (i.e. nematodes, copepods, amphipods, polychaetes and oligochaetes), except for isopods. Starting at concentrations of 200 and 300 ng/l BaP, respectively, significant changes were observed at community level. At taxonomic level, the nematode communities were dominated at the start of the experiment and also after being exposed or not to BaP by Odontophora villoti, explicable through its high ecologic ubiquity and the presence of well-developed chemosensory organs (i.e. amphids), which potentially increased the avoidance reaction following exposure to this hydrocarbon. Moreover, changes in the activity of several biochemical biomarkers (i.e. catalase 'CAT', gluthatione S-transferase 'GST', and ethoxyresorufin-O-deethylase 'EROD') were observed in the nematode species Oncholaimus campylocercoides, paralleled by significant decreases in CAT activity for non-gravid females compared to controls at concentrations of 25 ng/l BaP and associated with significant increase in GST and EROD activities for both types of individuals.

Pseudomonas spp. Mediate defense response in sugarcane through differential exudation of root phenolics.

Pseudomonas spp., a ubiquitous biocontrol agent, protects the plants from phytopathogens by suppressing them directly by reinforcing the plant's intrinsic defense mechanism. Root exudated phenolics play an important role in establishing the rhizobacteria population and cross the host boundaries in beneficial plant-microbe interaction. In this study, Pseudomonas spp. HU-8 & HU-9 antagonized the sugarcane red rot pathogen (C. falcatum) and showed a positive chemotactic response against different concentrations (10-30 µM) of synthetic phenolic acids like p-coumaric, vanillic, and 3,4 di-hydroxybenzoic acid. In a pot experiment, they effectively colonized the sugarcane rhizosphere and mediated defense response in sugarcane plants challenged with red rot pathogen C. falcatum by regulating the exudation of root phenolics under hydroponic conditions. They significantly induced the activity of the antioxidant enzymes CAT (1.24-1.64 fold), PO (0.78-1.61 fold), PAL (0.77-0.97 fold), and PPO (3.67-3.73 fold) over untreated plants in sugarcane. They also induced the total phenolic contents (TPC) in sugarcane in the presence (6.56-10.29 mg/g GAE) and absence (2.89-4.16 mg/g GAE) of the pathogen quantified through the Folin-Ciocalteu (FC) method. However, their effect was lower than that of the pathogen (4.34-8 mg/g GAE). The Pseudomonas spp. significantly colonized the sugarcane rhizosphere by maintaining a cell population of (1.0E + 07-1.3E + 08 CFU/mL). A significant positive Pearson's correlation was observed between the root exudated total phenolic contents, antioxidant enzymatic activities, and rhizospheric population of inoculated bacteria. The 16S rRNA and rpoD gene analysis showed sequence conservation (C: 0.707), average number of nucleotide differences (k: 199.816), nucleotide diversity, (Pi): 0.09819), average number of informative nucleotide sites per site (Psi: 0.01275), GC content (0.57), and polymorphic sites (n = 656). These diverse Pseudomonas spp. could be an ideal bio-inoculants for a broad range of hosts especially graminaceous crops.

Bacterial Endophyte Community Dynamics in Apple (Malus domestica Borkh.) Germplasm and Their Evaluation for Scab Management Strategies.

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.

Effect of potassium fertilizer on the growth, physiological parameters, and water status of Brassica juncea cultivars under different irrigation regimes.

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.

Methyl Jasmonate and Sodium Nitroprusside Jointly Alleviate Cadmium Toxicity in Wheat (Triticum aestivum L.) Plants by Modifying Nitrogen Metabolism, Cadmium Detoxification, and AsA-GSH Cycle.

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 (F v /F m ), 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 FINDINGS: The 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.

Multi-biomarker approach in Mytilus galloprovincialis and Ruditapes decussatus as a predictor of pelago-benthic responses after exposure to Benzo[a]Pyrene.

This study evaluated the biomarker responses indicative of exposure to Benzo[a] Pyrene (B[a]P) in Mytilus galloprovincialis and Ruditapes decussatus. A significant increase of the total oxyradical scavenging capacity (TOSC) was observed after seven days of exposure to two concentrations of B[a]P (100 and 300 µg.L-1), in the digestive gland with the lowest concentration tested. The TOSC in the gills increased notably only after the exposure to 300 µg.L-1 of B[a]P. Interestingly, the superoxide dismutase (SOD) and catalase (CAT) activities in gills and digestive gland on one hand and glutathione S-transferase (GST) in gills in the other, were positively correlated with the concentration of B[a]P with a significant induction noticed at the highest concentration. In contrast, a significant increase of the GST activity was observed in the digestive gland following the exposure of bivalves to 100 µg.L-1. In pelagic (M. galloprovincialis) or benthic (R. decussatus) bivalves, the AChE activity decreased discernibly in digestive glands and gills with the increase of B[a]P concentrations as evidence of neurotoxic effects. In clams, the exposure to B[a]P was followed by a significant increase of Malondialdehyde level (MDA) in gills and digestive gland, this does not occur in gills of Mytilus galloprovincialis at the concentration of 100 µg.L-1. Overall, the results found seems to indicate that the mussel was more suitable as a predictor tool of toxicity of B[a]P.