Response surface optimization of cellulase production from Aneurinibacillus aneurinilyticus BKT-9: An isolate of urban Himalayan freshwater.

Due to their vast industrial potential, cellulases have been regarded as the potential biocatalysts by both the academicians and the industrial research groups. In the present study, culturable bacterial strains of Himalayan Urban freshwater lake were investigated for cellulose degrading activities. Initially, a total of 140 bacterial strains were isolated and only 45 isolates were found to possess cellulose degrading property. On the basis of preliminary screening involving cellulase activity assay on CMC agar (with clear zone of hydrolysis) and biosafety assessment testing, only single isolate named as BKT-9 was selected for the cellulase production studies. Strain BKT-9 was characterized at the molecular level using rRNA gene sequencing and its sequence homology analysis revealed its identity as Aneurinibacillus aneurinilyticus. Further, various physico-chemical parameters and culture conditions were optimized using one factor approach to enhance cellulase production levels in the strain BKT-9. Subsequently, RSM based statistical optimization led to formulation of cellulase production medium, wherein the bacterial strain exhibited ~60 folds increase in enzyme activity as compared to un-optimized culture medium. Further studies are being suggested to scale up cellulase production in A. aneurinilyticus strain BKT-9 so that it can be utilized for biomass saccharification at an industrial level.

Biological Efficacy of Essential Oils and Plant Extracts of Cultivated and Wild Ecotypes of Origanum vulgare L.

Current study describes discrepancy in biological efficacy of methanolic and ethanolic extracts and essential oil procured from cultivated and wild accessions of Origanum vulgare. Simultaneously, quantification of carvacrol, thymol, caryophyllene, ocimene, and terpinen-4-ol contents was determined via GC-MS and GC in both accessions. The results revealed significantly a higher antioxidant potential by methanolic extracts displaying IC50 of 19.9 µg/ml compared to essential oil with IC50 of 10 µg/ml, and ethanolic extracts were found to be less effective even at the concentration of 3 µg/ml. However, essential oil from wild and cultivated accessions of O. vulgare exhibited significantly high antimicrobial activity against all 39 bacteria, 16 fungi, and 2 yeast species tested due to higher concentrations of carvacrol and thymol as revealed by GC analysis. Inhibition of tyrosinase activity in a C6 cell line displayed 81.0%-87.0% depigmentation potential of the methanolic extracts, while ethanolic extracts revealed a maximum of 88.54-99.02% inhibition of reactive oxygen species (ROS) in H2O2-treated cells. Hence, the study determines efficacy of essential oil against microbial pathogenesis, methanolic extracts as potent depigmentation agents, and ethanolic extracts as potent free radical scavenger.

Isolation, purification and characterization of naturally derived Crocetin beta-d-glucosyl ester from Crocus sativus L. against breast cancer and its binding chemistry with ER-alpha/HDAC2.

Saffron plant (Crocus sativus L.) is being used as a source of saffron spice and medicine to cure or prevent different types of diseases including cancers. We report the isolation, characterization of bioactive small molecule ([crocetin (ß-d-glucosyl) ester] from the leaf biowastes of saffron plant of Kashmir, India. MTTC assay and Bio-autography aided approach were used to assess anti-oxidant activity and anti-cancer properties of crocin (s) against DPPH free radical and breast cancer cell line respectively. Crocetin beta-d-glucosyl ester restrained proliferation of human breast adeno-carcinoma cell model (MCF-7) without significantly affecting normal cell line (L-6). Further studies involving molecular mechanics generalized born surface area and molecular docking showed that crocetin beta-d-glucosyl ester exhibits strong affinity for estrogen receptor alpha and histone deacetylase 2 (crucial receptors involved in breast cancer signalling) as evidenced by the negative docking score and binding free energy (BFE) values. Therefore, crocetin beta-d-glucosyl ester from Crocus sativus biowastes showed antiproliferative effect possibly by inhibiting estrogen receptor alpha and HDAC2 mediated signalling cascade.

Elucidating genetic variability and population structure in Venturia inaequalis associated with apple scab diseaseusing SSR markers.

Apple scab caused by Venturia inaequalis Cooke (Wint.) is one the important diseases of trade and industrial significance in apple. In present study variability studies in pathogen isolates were studied, which is one of the most important factors for devising management studies of scab disease in apple. Genetic diversity of 30 Venturia inaequalis isolates from 12 districts of two geographical distinct regions of Jammu and Kashmir was calculated based on the allele frequencies of 28 SSR markers and the internal transcribed spacer (ITS) region of the ribosomal DNA. The ITS based characterized sequences were submitted to NCBI GenBank and accession numbers were sanctioned. Dendrogram showed that all the accessions formed 2 main clusters with various degree of sub clustering within the clusters. Analysis based on SSR study reveals that the heterozygosity ranged from 0.0 and 0.5, with an average value of 0.39. The expected heterozygosis or gene diversity (He) ranged from 0.0 to 0.50 with an average of 0.40. The Fst value ranges from 0 to 0.6 with an average of 0.194. Diversity within each population (HS) values ranging from 0.26 to 0.33. Average differentiation among populations (GST) was 0.11 and populations were isolated by significant distance (r 2 = 0.50, P < 0.01). From the AMOVA analysis, 25% of variation was observed among population, 9% among individuals and 66% within individuals observed in the population. Structure analysis grouped isolates into two populations. Principle coordinate analysis explained variation of 36.6% in population 1, 14.30% in population 2 and 13.10% in population 3(Admixture) with 64.07% as overall cumulative percentage of variation. This indicates that extensive short-distance gene flow occurs in Kashmir region that dispersal over longer distances also appears to occur frequently enough to prevent differentiation due to genetic drift. Also it is evident that Jammu and Kashmir most likely has V. inaequalis subpopulations linked to diverse climatic conditions of the Jammu region compared to the mountainous inland Kashmir region. The results of present study would help to understand the genetic diversity of V. inaequalis from Jammu and Kashmir that would lead in the development of more effective management strategies and development of new resistant cultivars through marker-assisted selection.

Variability in Catechin and Rutin Contents and Their Antioxidant Potential in Diverse Apple Genotypes.

Catechins and rutin are among the main metabolites found in apple fruit. Sixty apple genotypes, harvested in 2016 and 2017, were analyzed for their phenolic content and antioxidant activity. The HPLC analysis showed that the catechin concentration ranged from 109.98 to 5290.47 µg/g, and the rutin concentration ranged from 12.136 to 483.89 µg/g of apple fruit. The level of DPPH activity ranged from 9.04% to 77.57%, and almost half of the 15 genotypes showed below 30⁻40% DPPH activity. The apple genotypes 'Lal Ambri', 'Green Sleeves', and 'Mallus floribunda' showed the highest DPPH activity of between 70% and 80%, while 'Schlomit', 'Luxtons Fortune', 'Mayaan', 'Ananas Retrine', and 'Chaubatia ambrose' showed the lowest ferric reducing antioxidant power (FRAP) activity (0.02⁻0.09%). Statistical analysis showed a correlation between DPPH activity and catechin content (r = 0.7348) and rutin content (r = 0.1442). Regarding antioxidant activity, fractionated samples of apple genotypes revealed significant activity comparable to that of ascorbic acid. There was also a consistent trend for FRAP activity among all apple genotypes and a significant positive correlation between FRAP activity and rutin content (r = 0.244). Thus, this study reveals a significant variation in antioxidant potential among apple genotypes. This data could be useful for the development of new apple varieties with added phytochemicals by conventional and modern breeders.

Roles of potential plant hormones and transcription factors in controlling leaf senescence and drought tolerance.

Plant leaves offer an exclusive windowpane to uncover the changes in organs, tissues, and cells as they advance towards the process of senescence and death. Drought-induced leaf senescence is an intricate process with remarkably coordinated phases of onset, progression, and completion implicated in an extensive reprogramming of gene expression. Advancing leaf senescence remobilizes nutrients to younger leaves thereby contributing to plant fitness. However, numerous mysteries remain unraveled concerning leaf senescence. We are not still able to correlate leaf senescence and drought stress to endogenous and exogenous environments. Furthermore, we need to decipher how molecular mechanisms of the leaf senescence and levels of drought tolerance are advanced and how is the involvement of SAGs in drought tolerance and plant fitness. This review provides the perspicacity indispensable for facilitating our coordinated point of view pertaining to leaf senescence together with inferences on progression of whole plant aging. The main segments discussed in the review include coordination between hormonal signaling, leaf senescence, drought tolerance, and crosstalk between hormones in leaf senescence regulation.

Variation in elemental composition as influenced by chlorpyrifos application in mung bean (Vigna radiata L.).

Chlorpyrifos (O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate), is an organophosphate insecticide effective against a broad spectrum of insect pests of economically important crops. The present study investigated the effects of chlorpyrifos application on sulfate assimilation and macro elemental composition in different plant parts at different phenological stages. Field experiments were conducted in the month of April 2008-2009. The individual plot size was 6 m2 (4 m × 1.5 m) having 4 rows with a row-to-row distance of 15 inches and plant to plant distance of 10 inches. The number of plants per m2 was 15. Seedlings were collected at 5 (preflowering), 10 (flowering) and 20 (postflowering) DAT (day after treatment) to analyze the effect of chlorpyrifos on APR activity and elemental composition. At harvest stage, seed from individual treatments were analyzed for sulfur containing amino acids like methionine and cysteine content. Twenty-day-old seedlings of Vigna radiata L. were subjected to chlorpyrifos at different concentrations ranging from 0 to 1.5 mM through foliar spray in the field condition. A significant increase (50% in cysteine content and 50-92% in methionine content) in sulfur containing amino acids at a higher dose rate of 1.5 mM was recorded in seeds, however the increased activity of adenosine 5-phosphosulfate reductase (APR), the key enzyme in sulfate assimilation was recorded in all the three parts of the plant (leaf, stem, root.). Transiently lower nitrogen, sulfur and carbon content in 0.6 and 1.5 mM chlorpyrifos application in V. radiata L. supports the inhibition of metabolic processes. However, reverse trend was exhibited at 0.3 mM for same parameters. These results suggest the stimulatory effects on sulfate assimilation in V. radiata L. while as inhibitory effects were prevalent on elemental composition.

Effect of environmental variables on phytonutrients of Origanum vulgare L. in the sub-humid region of the northwestern Himalayas.

Ecological and soil physiochemical parameters impact the crop quality and development. In spite of the huge commercial prospective, the phytonutrient and chemometric profiles of Himalayan oregano (Origanum vulgare L.) have not been evaluated, and their relationships with ecological parameters are still lacking. The objective of this research study was to evaluate the disparity in the phytonutrient profiles of different ecotypes of O. vulgare in wild and cultivated populations and determine whether such variation was related to the diverse climatic and edaphic conditions prevailing in the northwestern Himalayas. Micrometeorological, atomic absorption spectroscopy for micro-elemental analysis was determined for soil. HPLC was used to determine the disparity in phytonutrient (quercetin, betacarotene, ascorbic acid, and catechin) and phytochemical (arbutin) levels. Cultivated populations had lower phytonutrient levels than wild populations. The habitat exhibiting pH values ranging from 6 to 7 elevated organic carbon (2.42%), nitrogen (97.41 kg ha-1), and manganese (10-12 µg g-1) and zinc contents (0.39-0.50%) show luxirant growth of Origanum vulgarel. The phytonutrient (quercetin, betacarotene, ascorbic acid, arbutin, and catechin) levels had a direct relationship with UV-B flux (r2 = 0.82) and potassium (r2 = 0.97). Wild accessions predominantly contained catechin and ascorbic acid, with maximum values of 163.8 and 46.88 µg g-1, respectively, while the cultivated accessions had the highest level of arbutin (53.42 µg g-1). Maximum variation was observed in quercetin (114.61%) followed by ß-carotene (87.53%). Cultivated accessions had less quercetin (0.04-1.25 µg g-1) than wild accessions (1.25-2.87 µg g-1). Wild accessions had higher phytonutrient values for catechin, ß-carotene, and ascorbic acid while cultivated accessions had maximum values for arbutin. The correlation of environmental variables with phytonutrient levels paves the way for metabolomic-guided enhancement of agricultural practices for better herb quality.

Interactive effect of 24-epibrassinolide and silicon alleviates cadmium stress via the modulation of antioxidant defense and glyoxalase systems and macronutrient content in Pisum sativum L. seedlings.

BACKGROUND: This study assessed the effects of 24-epibrassinolide (EBL, 10-7M) and silicon (2 mM) on the alleviation of cadmium (Cd, 150 mg L-1) toxicity in Pisum sativum L. seedlings via the modulation of growth, antioxidant defense, glyoxalase system, and nutrient uptake. RESULTS: Shoot and root lengths declined by 46.43% and 52.78%, respectively, following Cd stress. Shoot and root dry weights also declined with Cd toxicity. Biochemical and physiological aspects exhibit significant decline including total chlorophyll (33.09%), carotenoid (51.51%), photosynthetic efficiency (32.60%), photochemical quenching (19.04%), leaf relative water content (40.18%), and gas exchange parameters (80.65%). However, EBL or Si supplementation alone or in combination modulates the previously mentioned parameters. Cadmium stress increased proline and glycine betaine (GB) contents by 4.37 and 2.41-fold, respectively. Exposure of plants to Cd stress increased the accumulation of H2O2, malondialdehyde content, electrolyte leakage, and methylglyoxal, which declined significantly with EBL and Si supplementation, both individually and in combination. Similarly, Cd stress adversely affected enzymatic and non-enzymatic antioxidants, but EBL and/or Si supplementation maintained antioxidant levels. Glyoxalase I (GlyI) accumulated after Cd stress and increased further with the application of EBL and Si. However, GlyII content declined after Cd stress but increased with supplementation of EBL and Si. Cadmium accumulation occurred in the following order: roots > shoots>leaves. Supplementation with EBL and Si, individually and in combination reduced Cd accumulation and enhanced the uptake of macronutrients and micronutrients in shoots and roots, which declined with Cd toxicity. CONCLUSION: The application of 24-EBL and Si, individually and in combination, alleviated the adverse effects of Cd by improving growth, biochemical parameters, nutrient uptake, osmolyte accumulation, and the anti-oxidative defense and glyoxalase systems in Pisum sativum seedlings.

Manipulation of Plant Growth Regulators on Phytochemical Constituents and DNA Protection Potential of the Medicinal Plant Arnebia benthamii.

Arnebia benthamii of the family Boraginaceae is a critically endangered nonendemic plant of the Kashmir Himalayas and is used to treat a number of human diseases. The current study was based on developing an in vitro micropropagation protocol vis-à-vis induction of various secondary metabolites under in vitro conditions for the possible biological activity. A tissue culture protocol was developed for A. benthamii for the first time in the Himalayan region using varied combinations and proper media formulations, including various adjuvants: Murashige and Skoog (MS) media, growth hormones, sugars, agar, and so forth. The influence of different media combinations was estimated, and the MS + thidiazuron (TDZ) + indole 3-acetic acid (IAA) combination favors a higher regeneration potential. The higher amounts of chemical constituents were also recorded on the same treatment. The in vitro plant samples also showed a noteworthy effect of scavenging of hydroxyl radicals vis-à-vis protection from oxidative DNA damage. The in vitro raised plants are good candidates for the development of antioxidant molecules.

Bioefficacy potential of different genotypes of walnut Juglans regia L.

The purpose of investigation was to assess the phytochemical and nutraceutical of walnut in leaf extracts through diverse quantitative and qualitative phytochemical tests followed by array of assays. The screening of 50 elite walnut genotypes which exhibited wide range of discrepancy in terms of phytochemicals as well as their anti-oxidant potential was done. Walnut genotypes displayed maximum divergence in quercetin content (2.86-5.78 mg/100 g) as represented by cluster analysis. The phenolic rich genotypes exhibiting total phenols (37.61-46.47 mg/g GAE) having higher DPPH potential (IP of 32.82-73.50) where as genotypes that accumulate flavonoids/flavanols (5.52-28.48 mg/g QE and 4.11-21.76 mg/g QE showed immense FRAP activity (418.92-1067.94 µM Fe2+/g FW). There was positive correlation between the phenolics content and anti-oxidant potential. The results showed oil content of 50.1-85.08% and kernel percentage 25.21-81.92% of all walnut genotypes. To evaluate the anti-proliferative potential of walnut genotypes, Trypan blue exclusion test, MTT assay and Griess assay was used. Each assay was repeated with different positive controls against a panel of human cancer cell lines viz THP-1, U2OS, IMR-32 and HBL-100 and then compared with the walnut extracts for their efficiency in anti-proliferative activity. The SPS 1 walnut extract at concentration of 500 µg/ml exhibited 10% cell viability and with 1000 µg/ml walnut extract there was consequent decline towards (6.25%) viability. The results indicated that walnut leaf constitutes an excellent source of effective natural antioxidants and chemo-preventive agents that can act as anti cancer agents.

Microclimatic variation in UV perception and related disparity in tropane and quinolizidine alkaloid composition of Atropa acuminata, Lupinus polyphyllus and Hyoscyamus niger.

The aim of current research was to evaluate the physiological adjustment in three medicinal herbs viz., Atropa acuminata, Lupinus polyphyllus and Hyoscyamus niger to the winter period characterised by intense UV flux in Kashmir valley across the North Western Himalaya. Quinolizidine (QA) and tropane alkaloid (TA) concentrations were analysed in these herbs thriving at two different altitudes via GC-MS and correlated by PCA analysis. This study investigated the hypothesis that UV reflectance and absorbance at low temperatures are directly related to disparity in alkaloid accumulation. Among QAs in L. polyphyllus, ammodendrine and lupanine accumulated at higher concentration and exhibited significant variation of 186.36% and 95.91% in ammodendrine and lupanine respectively in both sites. Tetrahydrohombifoline displayed non-significant variation of about 9.60% irrespective of sites. Among tropane alkaloid (TA), hyoscyamine was recorded as the most abundant constituent irrespective of the plant and site while apotropine accumulated in lesser quantity in A. acuminata than H. niger. However, apotropine demonstrated significant variation of 175% among both sites. The final concentration of quinolizidine (QA) and tropane alkaloid (TA) reflects the interplay between reflectance and absorbance of UV radiation response field. These findings suggest that spectral response of UV light contributes directly to alkaloid biosynthesis.

Selective Effect of Pesticides on Plant--A Review.

This review represents systematic and integrated picture of pesticide exposure to plant and its effect on growth and metabolism. Decades ago, agrochemicals were introduced aiming at enhancing crop yields and protecting crops from pests. Due to adaptation and resistance developed by pests to chemicals, every year higher amounts and new chemical compounds are used to protect crops, causing undesired side effects and raising the costs of food production. Biological chemical free agriculture is gaining also more and more support but it is still not able to respond to the need for producing massive amounts of food. The use of agrochemicals, including pesticides, remains a common practice especially in tropical regions and South countries. Cheap compounds, such as DDT, HCH, and Lindane, that are environmentally persistent, are today banned from agriculture use in developed countries, but remain popular in developing countries. As a consequence, persistent residues of these chemicals contaminate food and disperse in the environment. Therefore, the thrust of this paper was to review the application of pesticides effect early from germination to growth of the plant, leading to alteration in biochemical, physiological and different enzymatic and non-enzymatic antioxidants which ultimately affect the yield and resulted in residues in plant, vegetables, and fruits.

Differential response of terpenes and anthraquinones derivatives in Rumex dentatus and Lavandula officinalis to harsh winters across north-western Himalaya.

Herbs adapted to diverse climates exhibit distinct variability to fluctuating temperatures and demonstrate various metabolic and physiological adaptations to harsh environments. In this research, Rumex dentatus L. and Lavandula officinalis L. were collected before snowfall in September-November to evaluate variability in major phytoconstituents to diverse seasonal regime. LC-MS was used for simultaneous determination of eight anthraquinone derivatives in R. dentatus, i.e. emodin, physcion, chrysophanol, physcion glucoside, endocrocin, emodin glucoside, chrysophanol glucoside and chromone derivatives and monoterpenes in L. officinalis i.e. (Z)-ß-ocimene, (E)-ß-ocimene, terpene alcohol, terpin-4-ol, acetate ester-linalyl acetate and bicyclic sesquiterpene (E)-caryophyllene. The correlation analysis confirmed significant variation in anthraquinone glucoside and terpene content within Rumex and Lavender, respectively, and altitude was established as the determinant factor in secondary metabolism of both herbs. The study concludes the propagation of herbs in bioclimatic belts which favour accumulation of major constituents and validate their greater pharmacological activity.

Anti-oxidant modulation in response to gamma radiation induced oxidative stress in developing seedlings of Psoralea corylifolia L.

The seeds of Psoralea corylifolia L., an important medicinal herb in Indian and Chinese Pharmacopeia were exposed to gamma rays (2.5, 5, 10, 15 and 20 kGy) from Co(60) source at dose rate of 1.65 kGy h(-1). Enzymatic and non-enzymatic anti-oxidant responses were verified according to the developmental stages and gamma dose applied. Plants grown from seeds exposed to higher gamma doses exhibit higher activity of the antioxidants such as [Ascorbate peroxidase (APX, 1.11.1.1), superoxide dismutase (SOD, 1.15.1.1), glutathione reductase (GR, 1.6.4.2) and MDA content till flowering and declined thereafter. In contrast, CAT (1.11.1.6) activity declined in dose and age dependent manner. The correlation of gamma dose applied and oxidative stress was inferred from the increased enzymes activities and depression in total glutathione pool in seedlings developed from irradiated seeds. Nevertheless, the maintenance of high anti-oxidant capacity, psoralen accumulation seems to be an important strategy during acclimation of P. corylifolia to gamma radiation stress. Pronounced accumulation of psoralen following 15 and 20 kGy at post-flowering stage where oxidative stress is triggered modulates lipid peroxidation and proline accumulation. Further, in psoralen producing plants an increase in psoralen content can be used as a biomarker which specifies plant is under stress.

Enhancement in furanocoumarin content and phenylalanine ammonia lyase activity in developing seedlings of Psoralea corylifolia L. in response to gamma irradiation of seeds.

Gamma irradiation of seeds is known to be an important factor in stimulating biochemical and physiological processes. The aim of the present study was to investigate phenylpropanoids and associated enzymes responsible for the production of active metabolites. Furanocoumarin content was estimated in seeds of Psoralea corylifolia L. during two successive generations (G(1) and G(2)) where as phenylalanine ammonia lyase (PAL) activity was measured in leaves at different developmental stages of P. corylifolia L. raised from seeds irradiated with variable doses of gamma rays. Maximum accumulation of psoralen and isopsoralen was observed at 15 and 20 kGy doses during G(1) and G(2) generations, respectively. Psoralen proved to be the dominating metabolite in terms of its concentration, while isopsoralen was accumulated at relatively lower concentrations in successive generations. PAL activity was induced maximally following 15 and 20 kGy in G(1) plants and was preceded by psoralen and isopsoralen accumulation which peaked at the same dose rates in both generations. These effects were transmitted and prevalent in the next generation, that is, G(2) (indirectly irradiated). These long-term changes in plant metabolomics demonstrate genomic instability induced by gamma irradiation. However, no detrimental effects were seen at any irradiation dose in seeds. Furanocoumarin concentrations were also enhanced at 15 and 20 kGy. The present study further points out the persistence of changes in the biosynthesis of coumarin derivatives in the next generation. However, accumulation of these metabolites does not lead to any lethal effects.