Metabolomics reveals arbuscular mycorrhizal fungi-mediated tolerance of walnut to soil drought.

BACKGROUND: Arbuscular mycorrhizal fungi (AMF) have a positive effect on drought tolerance of plants after establishing reciprocal resymbiosis with roots, while the underlying mechanism is not deciphered. Metabolomics can explain the mechanism of plant response to environmental stress by analyzing the changes of all small molecular weight metabolites. The purpose of this study was to use Ultra High Performance Liquid Chromatography Q Exactive Mass Spectrometer to analyze changes in root metabolites of walnut (Juglans regia) after inoculation with an arbuscular mycorrhizal fungus Diversispora spurca under well-watered (WW) and drought stress (DS). RESULTS: Sixty days of soil drought significantly inhibited root mycorrhizal colonization rate, shoot and root biomass production, and leaf water potential in walnut, while AMF inoculation significantly increased biomass production and leaf water potential, accompanied by a higher increase magnitude under DS versus under WW. A total of 3278 metabolites were identified. Under WW, AMF inoculation up-regulated 172 metabolites and down-regulated 61 metabolites, along with no changes in 1104 metabolites. However, under DS, AMF inoculation up-regulated 49 metabolites and down-regulated 116 metabolites, coupled with no changes in 1172 metabolites. Among them, juglone (a quinone found in walnuts) as the first ranked differential metabolite was up-regulated by AMF under WW but not under DS; 2,3,5-trihydroxy-5-7-dimethoxyflavanone as the first ranked differential metabolite was increased by AMF under DS but not under WW. The KEGG annotation showed a large number of metabolic pathways triggered by AMF, accompanied by different metabolic pathways under WW and DS. Among them, oxidative phosphorylation and phenylalanine metabolism and biosynthesis were triggered by AMF in response to WW and DS, where N-acetyl-L-phenylalanine was induced by AMF to increase under DS, while decreasing under WW. CONCLUSION: This study provides new insights into the metabolic mechanisms of mycorrhiza-enhanced drought tolerance in walnuts.

Arbuscular mycorrhiza induces low oxidative burst in drought-stressed walnut through activating antioxidant defense systems and heat shock transcription factor expression.

Arbuscular mycorrhizal fungi (AMF) have important roles in enhancing drought tolerance of host plants, but it is not clear whether and how AMF increase drought tolerance in walnut (Juglans regia). We hypothesized that AMF could activate antioxidant defense systems and heat shock transcription factors (Hsfs) transcription levels to alleviate oxidative damage caused by drought. The walnut variety 'Liaohe No. 1' was inoculated with Diversispora spurca and exposed to well-watered (WW, 75% of the maximum soil water capacity) and drought stress (DS, 50% of the maximum soil water capacity) for 6 weeks. Plant growth, antioxidant defense systems, and expressions of five JrHsfs in leaves were studied. Such drought treatment inhibited root mycorrhizal colonization, while plant growth performance was still improved by AMF inoculation. Mycorrhizal fungal inoculation triggered the increase in soluble protein, glutathione (GSH), ascorbic acid (ASC), and total ASC contents and ascorbic peroxidase and glutathione reductase activities, along with lower hydrogen peroxide (H2O2), superoxide anion radical (O2 •-), and malondialdehyde (MDA) levels, compared with non-inoculation under drought. Mycorrhizal plants also recorded higher peroxidase, catalase, and superoxide dismutase activities than non-mycorrhizal plants under drought. The expression of JrHsf03, JrHsf05, JrHsf20, JrHsf22, and JrHsf24 was up-regulated under WW by AMF, while the expression of JrHsf03, JrHsf22, and JrHsf24 were up-regulated only under drought by AMF. It is concluded that D. spurca induced low oxidative burst in drought-stressed walnut through activating antioxidant defense systems and part Hsfs expressions.

Mycorrhizal Effects on Growth and Expressions of Stress-Responsive Genes (aquaporins and SOSs) of Tomato under Salt Stress.

Environmentally friendly arbuscular mycorrhizal fungi (AMF) in the soil can alleviate host damage from abiotic stresses, but the underlying mechanisms are unclear. The objective of this study was to analyze the effects of an arbuscular mycorrhizal fungus, Paraglomus occultum, on plant growth, nitrogen balance index, and expressions of salt overly sensitive genes (SOSs), plasma membrane intrinsic protein genes (PIPs), and tonoplast intrinsic protein genes (TIPs) in leaves of tomato (Solanum lycopersicum L. var. Huapiqiu) seedlings grown in 0 and 150 mM NaCl stress. NaCl stress severely inhibited plant growth, but P. occultum inoculation significantly improved plant growth. NaCl stress also suppressed the chlorophyll index, accompanied by an increase in the flavonoid index, whereas inoculation with AMF significantly promoted the chlorophyll index as well as reduced the flavonoid index under NaCl conditions, thus leading to an increase in the nitrogen balance index in inoculated plants. NaCl stress regulated the expression of SlPIP1 and SlPIP2 genes in leaves, and five SlPIPs genes were up-regulated after P. occultum colonization under NaCl stress, along with the down-regulation of only SlPIP1;2. Both NaCl stress and P. occultum inoculation induced diverse expression patterns in SlTIPs, coupled with a greater number of up-regulated TIPs in inoculated versus uninoculated plants under NaCl stress. NaCl stress up-regulated SlSOS2 expressions of mycorrhizal and non-mycorrhizal plants, while P. occultum significantly up-regulated SlSOS1 expressions by 1.13- and 0.45-fold under non-NaCl and NaCl conditions, respectively. It was concluded that P. occultum inoculation enhanced the salt tolerance of the tomato, associated with the nutrient status and stress-responsive gene (aquaporins and SOS1) expressions.

A Cross-Cultural Analysis of Plant Resources among Five Ethnic Groups in the Western Himalayan Region of Jammu and Kashmir.

Plant resources have always been valuable in human life, and many plant species are used in medicine, food, and ritual, and resource utilization is closely related to cultural diversity. Our study was conducted from June 2019 to April 2021, during which we aimed to document the local knowledge of plant resources of five ethnic groups, i.e., the Gujjar, Bakarwal, Kashmiri, Pahari, and Dogra communities of the Jammu and Kashmir (J&K) region, Western Himalayas. Through semi-structured interviews (N = 342) and group discussions (N = 38), we collected data on the ethnobotanical uses of plant resources. The data was subjected to hierarchical cluster analysis and ordination techniques (Principal Component Analysis) via, R software of version 4.0.0. Traditional uses were classified into three groups, i.e., single-, double-, and multi-use groups. The study recorded a total of 127 plant species, belonging to 113 genera and distributed among 64 botanical families. The dominant plant families were the Asteraceae, with 8% of all species, followed by Lamiaceae (6%), Polygonaceae (5%) and Ranunculaceae (4%). The recorded plant taxa were frequently used for medicine (51.4% responses), followed by food (14.9%), and fodder (9.5%). Principal component analysis (PCA) separated three groups of provisioning services depending on plant consumption preference levels. Comparative analysis showed remarkable similarities in plant uses (food, medicinal) among the Gujjar and Bakarwal ethnic groups, as both groups share a common culture. Some plants like Azadirachta indica, Brassica campestris, Ulmus wallichiana, Amaranthus blitum, and Celtis australis were also used for magico-religious purposes. We also recorded some medicinal uses that are new to the ethnobotanical literature of the J&K Himalayas, such as for Betula utilis, Sambucus wightiana, and Dolomiaea macrocephala, in our case for example local medicinal recipe, which is derived from Dolomiaea macrocephala, often known as Nashasta, used to treat weakness, back pain, and joint pain. Similarly, we also recorded new food uses for Eremurus himalaicus. Moreover, we also observed some plants for instance, Fragaria nubicola, Betula utilis and Juniperus communis have spiritual significance (i.e., amulets and scrolls) for this part of the Himalayan region. The present study provides a useful tool for resource management and can help in developing scientifically informed strategies for the conservation of plant resources.

Physiological and Biochemical Responses of Bicarbonate Supplementation on Biomass and Lipid Content of Green Algae Scenedesmus sp. BHU1 Isolated From Wastewater for Renewable Biofuel Feedstock.

In the present study, different microalgae were isolated from wastewater environment and evaluated for higher growth and lipid accumulation. The growth adaptability of all the isolated microalgae were tested for carbon source with supplementation of sodium bicarbonate in BG-11 N+ medium. Further based on the uptake rate of sodium bicarbonate and growth behavior, microalgal strains were selected for biofuel feedstock. During the study, growth parameters of all the isolates were screened after supplementation with various carbon sources, in which strain Scenedesmus sp. BHU1 was found highly effective among all. The efficacy of Scenedesmus sp. BHU1 strain under different sodium bicarbonate (4-20 mM) concentration, in which higher growth 1.4 times greater than control was observed at the concentration 12 mM sodium bicarbonate. In addition, total chlorophyll content (Chl-a + Chl-b), chlorophyll fluorescence (Fv/Fm, Y(II), ETR max, and NPQmax), and biomass productivity were found to be 11.514 µg/ml, 0.673, 0.675, and 31.167 µmol electrons m-2 s-1, 1.399, 59.167 mg/L/day, respectively, at the 12 mM sodium bicarbonate. However, under optimum sodium bicarbonate supplementation, 56.920% carbohydrate and 34.693% lipid content were accumulated, which showed potential of sodium bicarbonate supplementation in renewable biofuel feedstock by using Scenedesmus sp. BHU1 strain.

Root Endophytic Fungi Regulate Changes in Sugar and Medicinal Compositions of Polygonum cuspidatum.

Polygonum cuspidatum Sieb. et Zucc is an important industrial crop because it contains a large amount of medicinal secondary metabolites (such as polydatin, resveratrol, chrysophanol, and emodin). However, it is unclear whether root endophytic fungi increase the content of secondary metabolites in the plant. This study aimed to analyze the effects of Funneliformis mosseae (Fm) and Piriformospora indica (Pi) alone or in combination on plant growth, root morphology, thirteen sugars concentrations, and six secondary metabolites (physcion, chrysophanol, emodin, aloe-emodin, polydatin, and resveratrol) concentrations of P. cuspidatum. After 11 weeks of the fungal inoculation, the roots could be colonized by Fm and Pi single or in combination, along with the higher root colonization frequency of Fm > Pi > Fm + Pi in the descending order. In addition, Fm and Pi improved plant growth performance (plant height, stem diameter, leaf number, and shoot and root biomass) and root morphology (average diameter, maximum diameter, total length, area, and volume) to varying degrees, depending on fungal inoculations, in which Pi displayed a relatively better effect on plant growth. Single Fm and Pi inoculation significantly increased three disaccharides (sucrose, maltose, and trehalose) accumulation, while dual inoculum (Fm + Pi) only elevated sucrose concentrations. Most monosaccharides concentrations, such as D-arabinose, D-galactose, D-sorbitol, D-fructose, glucose, and L-rhamnose were not altered or inhibited by the endophytic fungi, except the increase in L-fucose and inositol. All fungal treatments significantly increased root chrysophanol and resveratrol concentrations, while decreased aloe-emodin concentrations. In addition, single Pi and dual Fm + Pi increased emodin concentrations, and single Fm and dual Fm + Pi elevated physcion and polydatin concentrations. It was concluded that Fm and Pi promoted the growth of P. cuspidatum, and the combination of Fm and Pi was more conducive to the production of some secondary metabolites than single inoculation.

Ethnoveterinary Practices of Medicinal Plants Among Tribes of Tribal District of North Waziristan, Khyber Pakhtunkhwa, Pakistan.

Domestic animals play a vital role in the development of human civilization. Plants are utilized as remedies for a variety of domestic animals, in addition to humans. The tribes of North Waziristan are extremely familiar with the therapeutic potential of medicinal plants as ethnoveterinary medicines. The present study was carried out during 2018-2019 to record ethnoveterinary knowledge of the local plants that are being used by the tribal communities of North Waziristan, Khyber Pakhtunkhwa, Pakistan. In all, 56 medicinal plant species belonging to 42 families were identified, which were reported to treat 45 different animal diseases. These included 32 herbs, 12 shrubs, and 12 trees. Among the plant families, Asteraceae contributed the most species (5 spp.), followed by Amaranthaceae (4 spp.), Solanaceae (4 species), and Alliaceae, Araceae, and Lamiaceae (2 spp. each). The most common ethnoveterinary applications were documented for the treatment of blood in urine, bone injury, colic, indigestion, postpartum retention, skin diseases, constipation, increased milk production, mastitis, foot, and mouth diseases.

Environmental variables drive plant species composition and distribution in the moist temperate forests of Northwestern Himalaya, Pakistan.

By assessing plant species composition and distribution in biodiversity hotspots influenced by environmental gradients, we greatly advance our understanding of the local plant community and how environmental factors are affecting these communities. This is a proxy for determining how climate change influences plant communities in mountainous regions ("space-for-time" substitution). We evaluated plant species composition and distribution, and how and which environmental variables drive the plant communities in moist temperate zone of Manoor valley of Northwestern Himalaya, Pakistan. During four consecutive years (2015-2018), we sampled 30 sampling sites, measuring 21 environmental variables, and recording all plant species present in an altitudinal variable range of 1932-3168 m.a.s.l. We used different multivariate analyses to identify potential plant communities, and to evaluate the relative importance of each environmental variable in the species composition and distribution. Finally, we also evaluated diversity patterns, by comparing diversity indices and beta diversity processes. We found that (i) the moist temperate zone in this region can be divided in four different major plant communities; (ii) each plant community has a specific set of environmental drivers; (iii) there is a significant variation in plant species composition between communities, in which six species contributed most to the plant composition dissimilarity; (iv) there is a significant difference of the four diversity indices between communities; and (v) community structure is twice more influenced by the spatial turnover of species than by the species loss. Overall, we showed that altitudinal gradients offer an important range of different environmental variables, highlighting the existence of micro-climates that drive the structure and composition of plant species in each micro-region. Each plant community along the altitudinal gradient is influenced by a set of environmental variables, which lead to the presence of indicator species in each micro-region.

Isolation and Characterization of Endophytes Bacterial Strains of Momordica charantia L. and Their Possible Approach in Stress Management.

In the present study, eight endophytic bacterial strains, namely Bacillus licheniformis R1, Bacillus sp. R2, Agrobacterium tumefaciens R6, uncultured bacterium R11, Bacillus subtilis RS3, Bacillus subtilis RS6, uncultured bacterium RS8 and Lysinibacillus fusiformis RS9, were isolated from the root of Momordica charantia L. All the strains, except R6 exhibited positive for IAA production, siderophore production, and phosphate solubilization during plant growth-promoting traits analysis. Strains invariably utilized glucose and sucrose as a carbon source during substrate utilization, while yeast extract, ammonium sulphate, ammonium chloride, glycine, glutamine, and isoleucine as nitrogen sources. In addition, Spectinomycin was found as the most effective during antibiotic sensitivity TEST, followed by Chloramphenicol, Erythromycin, Rifampicin and Kanamycin, while Polymixin B was found least effective, while strains R1, R6, and RS8 were sensitive to all the antibiotics. Strains R1 and RS6 were able to withstand tolerance up to 10% of NaCl. The strains showing resistance against broad-spectrum antibiotics, especially chloramphenicol, can be used in hospital waste management. In addition, strains with a tolerance of 10 % of NaCl can improve plant growth in the saline affected area.

Cloning of a CHS gene of Poncirus trifoliata and its expression in response to soil water deficit and arbuscular mycorrhizal fungi.

Flavonoids are secondary metabolites widely found in plants with antioxidants, of which chalcone synthase (CHS) is a key enzyme required in flavonoid synthesis pathways. The objective of this study was to clone a CHS gene from trifoliate orange (Poncirus trifoliata) and analyze its biological information and partial functions. A PtCHS gene (NCBI accession: MZ350874) was cloned from the genome-wide of trifoliate orange, which has 1156 bp in length, encoding 391 amino acids, with a predicted protein relative molecular mass of 42640.19, a theoretical isoelectric point of 6.28, and a lipid coefficient of 89.82. The protein is stable, hydrophilic, and high sequence conservation (92.49% sequence homology with CHS gene of other species). PtCHS was highly expressed in stems, leaves and flowers, but very low expression in roots and seeds. Soil water deficit could up-regulate expressions of PtCHS in leaves. An arbuscular mycorrhizal fungus, Funneliformis mosseae, significantly increased plant biomass production, CHS activity, expressions of PtCHS, and total flavonoid content in leaves and roots, independent of soil water status. Total flavonoids were significantly positively correlated with PtCHS expression in leaves only and also positively with root mycorrhizal colonization. Such results provide insight into the important functions of PtCHS in trifoliate orange.

Species Distribution Pattern and Their Contribution in Plant Community Assembly in Response to Ecological Gradients of the Ecotonal Zone in the Himalayan Region.

The ecotonal zones support populations that are acclimated to changing, fluctuating, and unstable conditions, and as a result, these populations are better equipped to adjust to expected change. In this context, a hypothesis was tested that there must be vegetation dominated by unique indicator plant species under the influence of ecological gradients in the ecotonal zone of Manoor Valley (northwestern Himalaya), Pakistan. Keeping the aforementioned hypothesis in mind, detailed field studies were conducted during different seasons in 2015-18. Line transect sampling and phytosociological characteristics (density, frequency, cover, and their relative values and Importance Value) were implemented as ecological methods. This investigation documented 97 plant species recorded from seven sampling sites. The community distribution modelling revealed that the ecological variables separate the seven sampling sites into two major plant communities (Indigofera-Parrotiopsis-Bistorta and Ziziphus-Leptopus-Quercus) recognized by TWINSPAN. The IBP communities showed a positive and significant correlation with altitude (1789.6-1896.3 m), sandy soil texture with a slightly acidic pH (6.4-6.5), and higher phosphorous (9-13 mg kg-1). In contrast with this, the ZLQ community was recognized on the southern slope under the strong influence of high electrical conductivity (2.82-5.4 dsm-1), organic matter (1.08-1.25%), calcium carbonate (5.8-7.6 mg kg-1), potassium (202-220 mg kg-1), and temperature (28.8-31.8 °C). Hence, both communities were found on opposite axes with clear differences based on the ecological gradients. NMDS clustered different species with similar habitats and different stands with common species, showing that plant species and stands were in a linear combination with ecological gradients. The IPB community has the maximum number of plant species (87 species), Shannon value (H' = 4), Simpson value (0.98), and Pielou's evenness value (0.96). Thus, the multivariate approaches revealed unique vegetation with sharp boundaries between communities which might be due to abrupt environmental changes.

Nigella sativa callus treated with sodium azide exhibit augmented antioxidant activity and DNA damage inhibition.

Nigella sativa L. (NS) is an herbaceous plant, possessing phytochemicals of therapeutic importance. Thymoquinone is one of the active phytochemicals of NS that confers noteworthy antioxidant properties. Sodium azide, an agent of abiotic stress, can modulates antioxidant system in plants. In the present investigation, sodium azide (0, 5 µM, 10 µM, 20 µM, 50 µM, 100 µM and 200 µM) doses administered to the in vitro NS callus cultures for production/modification of secondary metabolites with augmented activity. 200 µM sodium azide treated NS callus exhibited maximum peroxidase activity (1.286 ± 0.101 nanokatal mg-1 protein) and polyphenol oxidase activity (1.590 ± 0.110 nanokatal mg-1 protein), while 100 µM sodium azide treated NS callus for optimum catalase activity (1.250 ± 0.105 nanokatal mg-1 protein). Further, 200 µM sodium azide treated NS callus obtained significantly the highest phenolics (3.666 ± 0.475 mg g-1 callus fresh weight), 20 µM sodium azide treated NS callus, the highest flavonoids (1.308 ± 0.082 mg g-1 callus fresh weight) and 100 µM sodium azide treated NS callus, the highest carotenes (1.273 ± 0.066 mg g-1 callus fresh weight). However, NS callus exhibited a decrease in thymoquinone yield/content vis-à-vis possible emergence of its analog with 5.3 min retention time and an increase in antioxidant property. Treatment with 200 µM sodium azide registered significantly the lowest percent yield of callus extract (4.6 ± 0.36 mg g-1 callus fresh weight) and thymoquinone yield (16.65 ± 2.52 µg g-1 callus fresh weight) and content (0.36 ± 0.07 mg g-1 callus dry weight) and the highest antioxidant activity (3.873 ± 0.402%), signifying a negative correlation of the former with the latter. DNA damage inhibition (24.3 ± 1.7%) was recorded significantly maximum at 200 µM sodium azide treatment. Sodium azide treated callus also recorded emergence of a new peak at 5.3 min retention time (possibly an analog of thymoquinone with augmented antioxidant activity) whose area exhibits significantly negative correlation with callus extract yield and thymoquinone yield/content and positive correlation with antioxidant activity and in vitro DNA damage inhibition. Thus, sodium azide treatment to NS callus confers possible production of secondary metabolites or thymoquinone analog (s) responsible for elevated antioxidant property and inhibition to DNA damage. The formation of potent antioxidants through sodium azide treatment to NS could be worthy for nutraceutical and pharmaceutical industries.

The Effectiveness of Protected Areas in Conserving Globally Threatened Western Tragopan Tragopan melanocephalus.

Protected areas are a critical tool to conserve biodiversity in the face of the global crisis of species extinction. Here, we present the first ever management effectiveness assessment of Pakistan's Protected Areas (PAs). We link these assessments to the delivery of conservation outcomes focusing on the threatened Western Tragopan (Tragopan melanocephalus) endemic to Pakistan and India. We used two approaches, first mapping the spatial distribution of potential habitat coverage using machine learning ensemble models and second, an assessment of the management effectiveness of protected areas. Our results show that only Machiara National Park scored just above 40% (indicating relatively weak management), 22 of the PAs fell within the 25-50% quantile (indicating weak management), and 3 scored below 25% (indicating poor management). PAs within the species distributional range covered 92,387 ha which is only 2% of the total potential habitat of the Tragopan. Scoring of Planning element was insufficient both in term of the site and species. Likewise, inputs (e.g., research and monitoring program, staff numbers, staff training, current budget, security of budget, and management after process) were also inadequate. Finally, we recommend the establishment of more protected areas within the species potential habitat and inclusion of species-specific plans in Pakistan's PAs management.

Improvements in HOMA indices and pancreatic endocrinal tissues in type 2-diabetic rats by DPP-4 inhibition and antioxidant potential of an ethanol fruit extract of Withania coagulans.

CONTEXT: Withania coagulans (Stocks) Dunal fruits are used in the therapeutics of several ailments due to possessing of potent phytoconstituents which is also used traditionally for curing the diabetes. OBJECTIVE: The present study was assessing the amelioration potential of the phytochemicals of an ethanol fruit extract of W. coagulans (Stocks) Dunal in the HOMA (Homeostatic model assessment) indices and pancreatic endocrinal tissues by inhibition of DPP-4 and antioxidants activities. MATERIAL AND METHODS: The identification of phytoconstituents of the test extract was performed by LCMS. Further, assessments of in-vitro, in-vivo and in-silico were achieved by following standard methods. In-vivo studies were conducted on type-2 diabetic rats. RESULTS: The chosen extract inhibited DPP-4 activity by 63.2% in an in vitro assay as well as significantly inhibit serum DPP-4 levels. Accordingly, the administration of the ethanol fruit extract resulted in a significant (P ≤ 0.001) alterations in the lipid profile, antioxidant levels, and HOMA indices. Moreover, pancreatic endocrinal tissues (islet of Langerhans) appeared to have the restoration of normal histoarchitecture as evidenced by increased cellular mass. Molecular docking (Protein-ligands) of identified phytoconstituents with DPP-4 (target enzyme) shown incredibly low binding energy (Kcal/mol) as required for ideal interactions. ADMET analysis of the pharmacokinetics of the identified phytoconstituents indicated an ideal profile as per Lipinski laws. CONCLUSION: It can be concluded that the phytoconstituents of an ethanol fruit extract of W. coagulans have the potential to inhibit DPP-4 which result in improved glucose homeostasis and restoration of pancreatic endocrinal tissues in type-2 diabetic rats.

Nanoparticle-based amelioration of drought stress and cadmium toxicity in rice via triggering the stress responsive genetic mechanisms and nutrient acquisition.

Cadmium and drought are the most destructive of the abiotic stresses with negative consequences in terms of impaired metabolism, restricted nutrient use efficiency and disruptive photosynthesis of plants. The present study investigated the mitigation strategy of both aforementioned stresses by the application of iron oxide (IONPs) and hydrogel nanoparticles (HGNPs) simultaneously probably for the first time. IONPs were biofabricated by using a locally identified Bacillus strain RNT1, while HGNPs were produced chemically followed by the confirmation and characterization of both NPs through nanomaterials characterization techniques. Results of FTIR and XRD showed the capping of NPs by different functional groups together with their crystalline structure, respectively. SEM and TEM analysis showed the spherical shape along with the particle size ranging from 18 to 94 nm of both NPs, while EDS analysis confirmed the elemental purity of NPs. The results revealed that IONPs-treated rice plants increased biomass, antioxidant enzyme contents, photosynthesis efficiency, nutrient acquisition together with the decrease in reactive oxygen species and acropetal Cd translocation under normal and drought stress conditions as compared with control plants. Furthermore, the expression of the Cd transporter genes, OsHMA2, OsHMA3 and OsLCT1 were curtailed in NPs-treated rice plants under normal and drought stress conditions. The overall significance of the study lies in devising the NPs-based solutions of increasing heavy metal pollution and water availability challenges being faced the farmers around the world.

Classification and Characterization of the Manoor Valley's (Lesser Himalaya) Vegetation from the Subtropical-Temperate Ecotonal Forests to the Alpine Pastures along Ecological Variables.

Plant species are distributed in different types of habitats, forming different communities driven by different sets of environmental variables. Here, we assessed potential plant communities along an altitudinal gradient and their associations with different environmental drivers in the unexplored Manoor Valley (Lesser Himalaya), Pakistan. We have implemented various ecological techniques and evaluated phytosociological attributes in three randomly selected 50 m-transects within each stand (a total of 133) during different seasons for four years (2015-2018). This phytosociological exploration reported 354 plant species representing 93 different families. The results revealed that the Therophytic life form class dominated the flora, whereas Nanophyll dominated the leaf size spectra. There were a total of twelve plant communities identified, ranging from the lowest elevations to the alpine meadows and cold deserts. The maximum number of species were found in Cedrus-Pinus-Parrotiopsis community (197 species), in the middle altitudinal ranges (2292-3168 m). Our results showed that at high altitudes, species richness was reduced, whereas an increase in soil nutrients was linked to progression in vegetation indicators. We also found different clusters of species with similar habitats. Our study clearly shows how altitudinal variables can cluster different plant communities according to different microclimates. Studies such as ours are paramount to better understanding how environmental factors influence ecological and evolutionary aspects.

In Vivo Studies of Inoculated Plants and In Vitro Studies Utilizing Methanolic Extracts of Endophytic Streptomyces sp. Strain DBT34 Obtained from Mirabilis jalapa L. Exhibit ROS-Scavenging and Other Bioactive Properties.

Reactive oxygen species (ROS) and other free radicals cause oxidative damage in cells under biotic and abiotic stress. Endophytic microorganisms reside in the internal tissues of plants and contribute to the mitigation of such stresses by the production of antioxidant enzymes and compounds. We hypothesized that the endophytic actinobacterium Streptomyces sp. strain DBT34, which was previously demonstrated to have plant growth-promoting (PGP) and antimicrobial properties, may also have a role in protecting plants against several stresses through the production of antioxidants. The present study was designed to characterize catalase and superoxide dismutase (SOD), two enzymes involved in the detoxification of ROS, in methanolic extracts derived from six endophytic actinobacterial isolates obtained from the traditional medicinal plant Mirabilis jalapa. The results of a preliminary screen indicated that Streptomyces sp. strain DBT34 was the best overall strain and was therefore used in subsequent detailed analyses. A methanolic extract of DBT34 exhibited significant antioxidant potential in 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assays. The cytotoxicity of DBT34 against liver hepatocellular cells (HepG2) was also determined. Results indicated that methanolic extract of Streptomyces sp. strain DBT34 exhibited significant catalase and SOD-like activity with 158.21 U resulting in a 55.15% reduction in ROS. The IC50 values of a crude methanolic extract of strain DBT34 on DPPH radical scavenging and ABTS radical cation decolorization were 41.5 µg/mL and 47.8 µg/mL, respectively. Volatile compounds (VOC) were also detected in the methanolic extract of Streptomyces sp. strain DBT34 using GC-MS analysis to correlate their presence with bioactive potential. Treatments of rats with DBT34 extract and sitagliptin resulted in a significant (p ≤ 0.001) reduction in total cholesterol, LDL-cholesterol, and VLDL-cholesterol, relative to the vehicle control and a standard diabetic medicine. The pancreatic histoarchitecture of vehicle control rats exhibited a compact volume of isolated clusters of Langerhans cells surrounded by acinies with proper vaculation. An in-vivo study of Streptomyces sp. strain DBT34 on chickpea seedlings revealed an enhancement in its antioxidant potential as denoted by lower IC50 values for DPPH and ABTS radical scavenging activity under greenhouse conditions in relative comparison to control plants. Results of the study indicate that strain DBT34 provides a defense mechanism to its host through the production of antioxidant therapeutic agents that mitigate ROS in hosts subjected to biotic and abiotic stresses.

Biological soil crust (BSC) is an effective biofertilizer on Vigna mungo (L.).

Biological soil crusts (BSC) (Cyanobacteria) play an important role in the soil nitrogen fixation and soil stabilization. However, limited researches were carried out about the diversity and distribution of Bio crust in sacred forests. The study aims to identify the distribution of cyanobacteria in biological soil crusts from different sacred groves of Ariyalur and Pudukottai, Tamil Nadu. We identified following microbes of Microcoleus, Scytonema, Anabaena and Nostoc in biological soil crust. A surface experiment was conducted for the efficacy of biological soil crusts on crops seedling growths. The efficacy was assessed by measuring the root & shoot length, dry & fresh weight of the plant, total chlorophyll, protein & carbohydrate content, with reducing sugar. The plant growth was higher in biocrust inoculated pot than the control. In corresponding nitrogenase activity was determined by the acetylene reduction assay, and phytohormones documentation was executed by the high-performance liquid chromatography. The results indicating that the biological soil crusts are the promising factors influencing the plant growth by plant growth-stimulating auxins and nitrogenase activity.

Arbuscular mycorrhizal fungi modulates dynamics tolerance expression to mitigate drought stress in Ephedra foliata Boiss.

Arbuscular mycorrhizal fungi (AMF) are one of the most important drivers of soil ecosystem dynamics. AMF have the potential to improve plant growth and development by modulating key hormonal pathways, which result in decreasing the adverse impact of abiotic stress, such as drought. Pot experiments were conducted in this study to investigate the ability of AMF to ameliorate the adverse impact of drought in Ephedra foliate. Non-inoculated AMF E. foliate (Ef) plants, exhibited reduced growth in response to drought stress with a concomitant lowering of chlorophyll pigments, relative to non-stressed and AMF inoculated plant. AMF inoculated E. foliate showed improved nitrogen metabolism by positively regulating nitrate and nitrite reductase activity which results in greater ammonium availability for the synthesis of amino acids. Inoculation with AMF also increased antioxidant enzyme activity, ascorbic acid contents, and reduction in glutathione level. This resulted in significant amelioration of oxidative damage to plant membranes by restricting the excess generation of reactive oxygen species (ROS), such as hydrogen peroxide. Greater content of proline, glucose, and total soluble protein in AMF-inoculated plants provided further benefit to E. foliate plants and their ability to withstand drought stress, and also evident by a greater level of sucrose phosphate synthase activity. AMF significantly enhanced the uptake of essential nutrients like K, Mg, and Ca. Importantly, higher concentrations of plant hormones, including indole acetic acid (IAA), indole butyric acid (IBA), gibberellic acid (GA), and abscisic acid (ABA), were maintained in AMF-inoculated Ef plants. AMF inoculation also boosted phosphorous metabolism by increasing alkaline and acid phosphatase enzyme activity. In summary, AMF-inoculation of Ef plants significantly reduced the deleterious effect of drought stress by up-regulating the antioxidant defense system, synthesis of osmolytes, and maintaining phytohormone levels.

Arbuscular mycorrhizal fungi and biochar improves drought tolerance in chickpea.

Arbuscular mycorrhizal fungi (AMF) inoculation and biochar amendment has been reported to improve growth of several crop plants however their role in stress amelioration individually as well as in combination has not been worked out. This experiment was conducted to evaluate the application of AMF and biochar on the performance of chickpea under drought stress. The treatments included the individual as well as combined treatment of AMF and biochar to drought stressed and normal chickpea plants. Plants inoculation improved growth in terms of shoot and root length, leaf area and number of branches which was observed to show a steep decline due to drought stress. Drought declined the AMF colonization potential though biochar amendment ameliorated the negative effects of drought significantly by improving the spore population, number of mycelium, vesicle and arbuscules and the percentage of colonization as well. Increased chlorophyll synthesis in biochar and AMF treated plants was obvious, which lead to significant enhancement in the net photosynthetic efficiency. Drought stress also declined the relative water content (RWC) and membrane stability index (MSI), while treatment of biochar and AMF either individually or in combination mitigated the deleterious effects to considerable extent and caused a significant enhancement in RWC and MSI under normal conditions. Amendments with biochar and AMF inoculation increased the nitrogen fixation attributes including the number and weight of nodules, leghemoglobin content and activity of nitrate reductase enzyme leading to greater uptake and assimilation of nitrogen in them when compared to drought stressed plants. Drought stressed chickpea plants exhibited considerable reduction in uptake of nitrogen and phosphorous which was ameliorated by biochar and AMF treatments. It could be suggested that increase in growth and physiological attributes in chickpea due to biochar amendments and AMF inoculation under drought stress were plausibly due to their involvement in nitrogen and phosphorous uptake, chlorophyll synthesis and photosynthesis.