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Open Access
Biochemical Characterization And Protein Expression Analysis Of Common Bean Under Mineral Stress
(Sher-e-Kashmir University of Agricultural Sciences & Technology of Jammu, 2024-01-04) Farooq, Asmat; Sharma, Vikas
Common bean (Phaseolus vulgaris) is the most important legume for human consumption and majorly limited by reduced phosphorus levels in soil. Improving P use efficiency in legumes is a worldwide challenge and the present study was carried out on P stressed common bean to compare differences in the morphological, physiological, biochemical and structural parameters under in vitro conditions. The study was a multi-omic approach (such as proteomics, transcriptomics, phenomics, ionomics as well as metabolomics) to analyze the impact of mineral stress. The results indicated that mineral stress induces large disturbances inside the plant which causes profound metabolic changes (e.g., in photosynthesis, energy generation, cell wall composition, generation of secondary metabolites, free amino acids, antioxidants, osmolyte production) and leads to inhibition of plant growth and development. It was observed that there was significant variation in the morphological, physiological, biochemical as well as structural parameters of common bean shoot under P stress as compared to the plant growing in presence of P or control (P+). FTIR spectroscopy, SEM, SEM-EDX and fluorescence spectroscopy, assisted in validating the modifications in structural and metabolic composition of plant under P deficiency stress. The proteomic analysis was carried out using SDS-PAGE (gel based) as well as LC-MS (gel free) to identify the proteins that are responsible for providing adaptation to the plant under essential mineral deficiency stress. Candidate proteins identified may contribute shoot adaptation to P deficiency and may be useful for improvement of common bean. The two differentially identified shoot proteins - homocysteine methyltransferase and DDB1-CUL4 associated factor 1 like, are located in the cytoplasm and nucleus respectively. Both genes expressed 2.56 and 3.13 fold expression under P deficiency common bean shoot which validated their expression at the transcription level using qRT-PCR. To conclude, common bean is thus, an excellent model to understand the biology of mineral stress tolerance.
Open Access
Exploring Anticancer, Antioxidant And Antibacterial Potential Of Some Medicinal Plants Of Bhaderwah Region Of Jammu And Kashmir
(Sher-e-Kashmir University of Agricultural Sciences & Technology of Jammu, 2023-11-30) Thappa, Chandan; Sharma, Vikas
The present research work was carried out to evaluate in vitro anticancer, antioxidant, antibacterial efficacy of three medicinal plants namely Arisaemajacquemontii, Berberis lycium, Plectranthusrugosus, collected from village ChilliKelar, Bhaderwah, Jammu and Kashmir. The extracts of above mentioned medicinal plants were prepared in different solvents like n-hexane, chloroform, ethyl acetate, acetone, methanol and evaluated for total phenolic content, total flavonoid content, in vitro anticancer potential, antioxidant and antibacterial efficiency. The highest phenolic and flavonoid content was observed in the methanolic extract of B. lyciumroot part. The maximum antioxidant potential assayed via DPPH, FRAP, metal ion chelation, was displayed by the methanolic extract prepared from the root part of B. lycium. Similarly, methanolic extract of B. lyciumroot part displayed the lowest MIC value against all the test bacterial strains, viz., Staphylococcus aureus MTCC7443, Micrococcus luteus MTCC4821, Bacillus subtilis MTCC2389, Escherichia coli MTCC2127, Klebsiella pneumoniae MTCC7172. The root part of B. lyciumchloroform extract showed remarkable in vitro anticancer potential against six human cancer cell lines from five different tissues - Breast (MCF-7, MDA-MB-231), Colon (HCT-116), Lung (A-549), Pancreatic (MIA PaCa-2), Prostate (PC-3) viaSRB assay.The isolation for active ingredients was carried out from the chloroform extract of B. lyciumroot and the characterization was done by NMR (1H and 13C), mass spectroscopy that led to the isolation / characterization of two compounds namely Beta-sitosterol and Berberine. Further, berberine, at different molar concentrations, showed significant results against seven human cancer lines from five different tissues with IC50 values for MCF-7 (15.72±1.69 μM), MDA-MB-231(23.52±1.51 μM), HCT-116 (5.27±1.19 μM), A-549 (28.15±1.72 μM), MIA PaCa-2 (22.26±1.63 μM), PANC-1 (9.24±1.88 μM) and PC-3 (23.31±1.81 μM), where HCT-116, a human cancer cell line from colon origin, showed minimum IC50 value. Accordingly, a couple of mechanistic assays of berberine were performed on HCT-116 cell lines where apoptosis was detected using DAPI staining, reactive oxygen species and mitochondrial matrix potential assay. To conclude, berberine could serve as a lead molecule for the development of anticancer drugs and will provide great service to cancer patients especially with colon carcinoma.
Open Access
Biofortification Of Fruit Juices Using Probiotics And Its Effect On Biochemical, Physico-Chemical, Functional And Sensory Characteristics
(Sher-e-Kashmir University of Agricultural Sciences & Technology of Jammu, 2023-11-30) Roy, Proloy Sankar Dev; Sharma, Vikas
The present research work was carried out on biofortification of fruit juices using probiotics and its effect on biochemical, physico-chemical, functional and sensory characteristics.The fruit juices ofkaronda (Carissa carandas), phalsa (Grewiaasiatica), jamun (Syzygiumcumini), were used as a matrix for biofortification using probiotics. The results demonstratedan increasing trend in the phenolic and flavonoid profile. Thehighest total phenolic (771.01±1.37 mg GAE/ml) and flavonoid (394.09±2.66 mg QE/ml) contentwas shown by phalsa juice fermented by Saccharomyces cerevisaeas compared to control. The juice of karonda, fermented by S.cerevisae. displayed the highest free radical scavenging activity viaDPPH assay, whereasjamun juice,fermented byLactobacillusacidophilusshowed highest reducing power activity andin case of FRAP assay,phalsa juice fermented by S.cerevisae exhibited enhanced FRAP value.The highest α-amylase activity(84.20±1.87)was seen in karonda juice, fermented by S. cerevisaewhile the highest α-glucosidase inhibitory activity(84.45±1.28) was recorded by phalsa juice,fermentedby S. cerevisae. Moreover, highest anti-hypertensive activity was observed in karonda juice, fermented byS. cerevisae (89.88±0.79%).Effect of fermentation on various physico-chemical parameters viz., pH, total acidity, total soluble solids, total soluble sugar, reducing sugar, was determined andthe phalsa juice fermented by Lactobacilluscasei demonstrated the lowest pH value.The most significant rise in total acidity was observed in L. caseifermented phalsa juice. L. acidophilus fermentation in phalsa juice led to maximum increase in total soluble solids. The quantification of reducing sugar was conducted post-fermentation with probiotics and the study revealed that the level of reducing sugar was decreased that may be due the probiotics, feed on the sugars present in the juices. The phalsa juice displayed augmentation in its mineral composition.The potassium (K) content showed significant increase, whereas the increase was less prominent for zinc (Zn), as compared to non-fermented juice. Employing the hedonic scale to assess sensory attributes, most volunteers indicated a preference for both the color and odor of the juices.Furthermore, in vitro gastrointestinal digestion was also performed and the results displayed significant increasein TPC and TFC content of the three juices. To conclude, fermentation based biofortification of fruit juices can be used as an alternative natural mechanism for improvement of phytochemicals/ bioactive ingredients in phalsa, jamun and karonda.
Open Access
Bioaugmentation of Mushrooms for Production of Bioactive Proteins
(Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, 2023-02) Zahoor, Arjumand; Gupta, Moni
The present investigation was conducted to produce bioactive mushroom protein hydrolysates (MPHs) from three different locally grown mushrooms (Pleurotus florida, Agaricus bisporus and Calocybe indica) with a simple and cost-saving procedure of fermentation (microbial proteolysis) using Lactobacillus acidophilus and Lactobacillus brewis. Crude protein hydrolysates exhibited ACE inhibition (IC50 0.01 mg/ml) and α-amylase inhibitory (IC50 3.17 mg/ml) activities and showed foaming capacity (24.43%), oil absorption (148.33%), whitening index (344.02), water retention (15.02%), foaming stability (19.25%), emulsifying activity (127.31%), water absorption (177.93%) properties and browning index (24.44). External gelation method displayed high encapsulation efficiency (63%) and enhanced functional characteristics of mushroom protein hydrolysates. The external gelation beads with 7% alginate were harder and had a maximum force of around 2000-3500g when analysed using texture analyser. Encapsulation enhanced thermal properties of mushroom protein hydrolysates as enthalpy increased (-3.97 and -255.53J/g) upon encapsulation. FTIR analysis confirmed an effective ionic complexation between sodium alginate and peptides present in the hydrolysates. Encapsulation significantly reduced the simulated gastric and intestinal digestibility of protein hydrolysate and increased its bioavailability. The study indicated that fermentation based mushroom protein hydrolysates had remarkable bioactive and functional properties and external gelation method enhanced the functional properties and bioavailability. MPH can be utilized as protein supplement loaded with bioactive peptides of low commercial value in food industry.
Open Access
Studies on Polymer Based Nanostructures for Controlled Release of Essential Oils, Their Characterization and Bioactive Properties
(Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, 2023-02) Majeed, Aasiya; Guleria, Sanjay
The present studyentitled, “Studies on polymer based nanostructures for controlled release of essential oils, their characterization and bioactive properties” was carried out with the objective to determine chemical composition of essential oils and estimate synergistic antimicrobial and antioxidant activity of essential oil combinations of four plants namely;Thymus serphyllum, Lavendula angustifolia, Rosemarinus officianalis and Pelargonium graveolens by GC-MS analysis; evaluation of their antioxidant, antimicrobial activity alone and/or in combination with each other or synthetic antibiotics (chloramphenicol ampicillin, erythromycin and amoxicillin); synthesis of different nanostructures; nano-encapsulation of essential oils in nanostructures and determination of their encapsulation efficiency, loading capacity, release kinetics; characterization of nanostructures using SEM, TEM and FTIR; In vitro evaluation of antimicrobial and antioxidant activity of nano-encapsulated essential oils. The major compounds identified in the EOs of Thymus serphyllum, Lavendulaangustifolia, Rosemarinus officinalis,and Pelargonium graveolens were o- cymene (50.49%), linalyl acetate (40.54%), camphor (36.54%) and citronellol (26.22%) respectively. Antioxidant studies indicated that the essential oil of Thymus serphyllum showed the highest DPPH radical scavenging activity (IC50 value of 74 ±3.15 µg/mL), reducing power (IC50 value of 831.33 ± 10.6 µg/mL) and FRAP activity (IC50 value of 106.77±1.75mM Fe2+eq./100mg). Thymus serphyllum EO also showed significant antimicrobial activity with MIC and MBC values ranging between 86 to 130 µg/mL and 95 to 165µg/mL against the test bacterial strains. Essential oil combinations were also evaluated for synergistic antioxidant and antimicrobial activities. In addition, essential oils in combination with antibiotics were also tested for synergistic antimicrobial activity. Furthermore, antimicrobial synergistic interactions were confirmed by time-kill kinetics studies. Chitosan-phenolic acid nanogels, chitosan-tripolyphosphate nanoparticles, and chitosan-alginate nanocapsules were also synthesized and encapsulated with test essential oils. The encapsulation efficiency ranged from 46.5% to 88.2%, 60.01% to 73%, and 68.33% to 78.58% while loading capacity varied from 16.48% to 23.76% and 18.53% to 24.20% for nanogels, nanoparticles, and nanocapsules respectively for test essential oils.Morphological analysis of nanostructures was carried out using SEM/TEM analysis and revealedthe spherical shape of nanogels with 100 nm size, oval shape in the case of nanocapsules with an average size of ≤ 1µm while spherical to the oval shape for chitosan nanoparticles with an average size of 40.66 nm. Similarly, nanostructures were also characterized by using FTIR, XRD, and DLS analysis.
Open Access
Biological activities of proteins and peptides from edible mushrooms
(Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, 2022-11) Nissa, Tabasum Un; Gupta, Moni
In the present study, firstly in silico approach was used to enzymatically hydrolyse the mushroom species for identification of peptide sequences in five edible mushrooms with antidiabetic (AntiDMPpred) and ACE (Antihypertensive peptide predictor) properties. WL (Trp-Leu) from Pleurotus ostreatus was found to be the most potent ACE inhibitory peptide with IC50 10.00 (μmol/L) and LGIADATPEQIALLLEDAATLLRPR peptide sequence from Agaricus bisporus showed higest antidiabetic probability of 0.82. Out of the five different mushrooms studied, the two best common edible mushrooms, Pleurotus ostreatus (fresh and dry) and Agaricus bisporus(fresh and dry) were chosen for further study. For optimization of degree of hydrolysis, the crude protein was extracted from the over matured mushroom which was further concentrated by 10% TCA and hydrolyzed using different concentration of papain (15,20,25,30 and 35 %), pH (6, 6.5 and 7) and time (6, 12, 18 and 24 hour) at constant temperature of 50 °C. Highest degree of hydrolysis 33.54 % was found in the sample of Agaricus bisporus (dry protein concentrate) at pH 6.5, enzyme concentration 30% and time 18 hrs and their concentrates and hydrolysates were studied for functional and biological properties like water activity which ranged from 0.63-0.31, emulsifying activity (104.66-145.83%), emulsifying stability (100.66-144.80%), fat absorption capacity (133.05-190.56%), colour (whitening and browning index) water absorption capacity (79.66-544%), total soluble proteins (16.24-290.35 mg/g), alpha amylase inhibition (IC50 3.38-14.33 mg/ml), alpha glucosidase inhibition (IC50 3.00-10.79 mg/ml) and ACE activities (IC50 0.19-9.64 mg/ml). Current investigation showed the soluble protein content in Pleurotus ostreatus and Agaricus bisporus ranged from of 16.24 to 277.88 and 20.04 to 290.35 mg/g and crude protein (19.62-50.10% and 22.38-55.53%) respectively. In-vitro gastrointestinal digestion (GID) study was done to check its effect on modulating the bio accessibility of the bioactive proteins and peptides. Highest degree of hydrolysis (52.80%) was observed in Agaricus bisporus dry protein concentrate at gastric phase. These hydrolyzed samples were profiled by running the SDS-PAGE with 12% resolving and 5% stacking gel;which showed banding pattern of 6.5 to 3.5 kDa. These bioactive peptide sequences and molecular weight of these compounds were quantified through MALDI-TOF-MS that showed the range of molecular weight in between 1262.25 to 1920.51 Da. The study showed that mature mushrooms can aid in developing value added nutraceutical products with beneficial bioactive proteins and peptides.
Open Access
Exploring antioxidant and antibacterial efficacy of medicinal plants from Kandi belt of Jammu region
(Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, 2023-01) Sharma, Saiyana; Sharma, Vikas
The present investigation was carried out to evaluate the antioxidant and antibacterial potential of some medicinal plants from Jammu region along with phytochemical composition analysis.Eleven (11) medicinal plants namely Adhatoda vasica, Artocarpus heterophyllus, Azadirachta indica, Calotropis procera, Cordia dichtoma, Emblica officinalis, Murraya koenigii, Syzygium cumini, Tinospora cordifolia, Terminalia chebula and Zanthoxylum armatumwere collectedfrom the Kandi belt of Jammu region andmethanolic extracts of the above-mentioned plants were prepared. The phytochemical analysis was carried out by determining the total phenolic and flavonoid content whereas the antioxidant activity was determined via three different assays like DPPH radical scavenging,ferric reducing antioxidant power and metal ion chelation assay.Furthermore, the antibacterial efficacy was tested by agar well diffusion method against five bacterial test strains viz., Bacillus subtilis MTCC2389, Micrococcus luteus MTCC4821, Staphylococcus aureus MTCC7443, Klebsiella pneumoniae MTCC7172, Escherichia coli MTCC2127 with minimum inhibitory concentration by broth dilution method. The results demonstrated that all the selected medicinal plants possess phenolic and flavonoid content, with Tinospora cordifolia commonly known as giloye showed highest phenolic and flavonoid content with values 350.19 ± 0.62mg GAE/g dw and 205.52 ± 0.64 mg CE/g dwrespectively. Similarly, all the medicinal plants under study displayed antioxidant potential, but T. cordifolia showed remarkable antioxidant potential with IC50 value 22.67 ± 0.57 µg/ml for DPPH assay, 3764.45 ± 1.46 μM Fe2+eq./g for FRAP assay and 69.39 ± 0.72 % for metal ion chelation assay. As per antibacterial studies are concerned, again T. cordifolia showed the maximum activity with least MIC against all the five bacterial strainsused in the current study. To conclude, the most active plant of our research work i.e.,T. cordifolia forms a good basis for its further phytochemical and pharmacological evaluation.
Open Access
Bromatological potential of faba bean seeds for application in food industry
(Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, 2022-11) Sharma, Sumit; Gupta, Moni
The present study was undertaken with the objective of evaluating the proximate and nutrient composition along with the antinutritional properties of faba bean seeds on dry weight basis. Thirty fababean (Vicia faba L.) germplasms were analyzed and the results revealed the presence of highest crude fibre in the germplasmEC-117741 (5.73±0.10%). The germplasmEC-243529 revealed the presence of highest percentage of crude protein (38.90±0.18 %) and EC-117842 had the highest fat content of 5.90±0.25 %. Analysis of total ash content showed highest percentage in germplasm EC-117739 with 6.45±0.27 % while highest energy value of 1598.78±6.90 (kJ/100g) was observed in the germplasm EC-117842. The germplasm EC-243529 was found to have 6.21 %total nitrogen which was the highest for any germplasm under study and EC-329668 showed the highest carbohydrate percentage (61.29 %). The highest protein denaturation activity was seen in germplasm RJ (39.58 %) followed by EC-267625(31.25%). Moreover it was seen that as the concentration of extract was increased the inhibition rate of egg albumun denaturation also increased. The highest linoleic acid was seen in germplasm RB (47.15%) The highest oleic acid, palmitic acid, stearic acid& linolenic acid was seen in germplasmsEC-628948 (32.1%), EC-1072(17.2%), KA (3.55%)&RE (5.1%)respectively. Anti nutritional factors viz. phytate contentwas lowest ingermplasmRSPaccounting to 4.50±0.13(mg/g) and minimum trypsin inhibitory activity in germplasmKA 0.74±0.02(mg/g). Among the exotic germplasms, the highest energy value was observed in EC-117842 and in RE germplasm 1578.12±5.02 (kJ/100g) among local germplasms. The study revealed that both exotic and local germplasms are excellent sources of nutritional components. All the germplasms showed a varying trend in their nutritional as well as anti nutritional properties due to collection from different site and their respective genetic makeup. The study will help in faba bean improvement for pulse breeding programme and also for utilization in food industry.
Open Access
Biochemical and molecular analysis of anthocyanins in common bean (Phaseolus vulgaris L.)
(Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, 2022-11) Singh, Amrinder; Gupta, Moni
The present investigation wascarried out to study anthocyanin profile of common bean germplasm and to check the differential expressed anthocyanin responsive genes in seeds grown under temperate and sub-tropical climate. 48common bean germplasm selectedwere diverse in seed colori.e black, dark red, red, brown, painted lady, frenchyellow and creamish white. Gene derived markers(SSR) markers were used to study anthocyanin diversity.Higher total anthocyanin, phenol, flavonoid content and DPPH activity were observed in seed coat than whole seed in both locations.SDSPAGEofseed proteinsexpoundednewpolypeptiderangingfrom166.9 to 6.84 KDa in RHRSS, Bhaderwah location whereas seed protein polypeptides from 207.09 to 13.41 KDa of SKUAST Jammu location as in the order, Delphinidin-3,5-di-O-glucoside> Results of UPLC-MS profile of anthocyanin dark red coloredgermplasm>Delphinidin-3-β-D-glucoside > Pelargonidin > Malvidin-3-O-glucoside > Peonidin-3-O-glucoside >Cyanidin-3-O-rhamnoside grown in two different location.To explicate the anthocyanin biosynthetic gene (ABG) pathway in Phaseolus vulgaris L. comparative genomic analysis between Arabidopsisthaliana, Glycine max L. and Phaseolus vulgaris L. was conducted on a genome level. 74 genes in Phaseolus vulgaris L. as orthologs of 41 anthocyanin biosynthetic genes in A. thaliana and Glycine max L. A total of 370 genic SSR primers and 74 wereidentified genes in Phaseolus vulgaris L. 16 primers were synthesized and 6 were found to be polymorphic among populations. A total of 43 alleles ranging from 2(BM 152) to 4(BM 157) were amplified with an average of 3.07 alleles per SSR, PIC value varied from 0.64 (BMb742) to 0.87 (BM159) with an average of 0.71.2D protein gel electrophoresis from seeds of BR 104 and BR 39 reported a total of 31 spots were differentially expressed {18 spots in BR 104(Bhaderwah) and 14 in BR 104(SKUAST-Jammu)} and 16 spots were differentially expressed{9 spots in BR 39 (Bhaderwah) and 7 in BR 39(SKUAST-Jammu)}.Thus, the study have generated enough useful data giving the insight in to identify high anthocyanin contents genotypeand the differential expressive anthocyanin responsive genes product of common bean seeds grown under temperate and sub-tropical climate which can be exploited for common bean breeding programme.

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