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Open Access
In vitro establishment of Polygonatum verticillatum (L.) All. and augmentation of its phytochemical profile using elicitors
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2017-11) Tiwari, Tanuja; Chaturvedi, Preeti
lants as a valuable and safe natural source of medicine have drawn much scientific and commercial attention in past few decades. Polygonatum verticillatum (L.) All., an Asparagaceae member, is an important and highly valued ‘Astavarga’ medicinal herb of North West Himalaya. Regeneration of the plant through seeds is a slow process in nature owing to epicotyl dormancy. Therefore, in the present study, an effort was made to break seed dormancy, increase seed germination potential and micropropagate P. verticillatum using epicotyl and sprouted buds. Among all storage conditions and PGRs, cold moist stratification of seeds (4±10C for 4 weeks) together with GA3 (100 mg/L) pretreatment followed by germination on GA3 (50 to 100 mg/L) supplemented MS medium was found most effective to improve seed germination. Maximum per cent germination (78±3.74) was achieved through elicitation using 50µM sodium nitroprusside (SNP). Maximum per cent bud induction (90±6.1) with maximum number of buds (7.0±0.45) and shoot length (7.10±3.18cm) was achieved from epicotyl explants inoculated in MS+TDZ(1.0mg/L)+NAA(0.5mg/L)+GA3(2.0mg/L). Microshoots inoculated in MS+KN (1.0mg/L)+IAA(0.5mg/L) and MS+IBA(1.0 mg/L) respectively showed maximum per cent microrhizome induction (95±5.0) and root induction (85±6.12). Sprouted bud explants inoculated in MS+TDZ (2.0mg/L)+NAA(1.0mg/L)+GA3(2.0mg/L) showed maximum per cent shoot induction(85±10.0). However, maximum per cent rooting (70±5.0) was achieved in MS+IBA (1.0 mg/L) and MS+NAA (1.0 mg/L)+KN(0.5mg/L). Plants raised through epicotyls and sprouted buds showed 80% and 56% survival rate respectively in garden soil. An effort was also made to investigate the effect of various chemical elicitors on phytochemical profiling of the plant. Maximum total chlorophyll content (1.67±0.05 mg g-1 fresh wt.), total phenolic content (14.71±0.47mg GAE g-1), total flavonoid content (14.66±0.17 mg QE g-1) and %DPPH radical scavenging activity (60.23±0.77) were achieved in SA (300 µM), JA (300 µL/L), SA (300 µM) and SNP (50µM) elicited plants respectively. Protein content (2.40±0.06 mg g-1 fresh wt.), peroxidase activity (1.33±0.03 unit min-1mg-1protein) and catalase activity (1.32±0.05 unit min-1mg-1protein) were augmented maximum in underground parts of JA (300 µL/L) and SNP (50 µM and 100 µM) elicited plants respectively, whereas SOD activity (4.79±0.13 unit min-1mg-1protein) was reported maximum in aerial parts of SA (300 µM) elicited plants. GC-MS analysis of differently elicited plants divulged remarkable (%) increase in medicinally important compounds. Diosgenin (anticancerous, antidiabetic), 9,12Octadecadienoic acid (z,z) (anti-inflammatory and antiarthritic), Gamma-sitosterol (antidiabetic) and Squalene (anticancerous, anti-tumor) increased to maximum viz., 68.11%, 89.67%, 78.87% and 81.82% respectively in SA treated plants. Guanosine (antiprotozoal), Palmitic acid (antioxidant), Vitamin E (antiinflammatory), 1,2-Benzenedicarboxylic acid (anti-inflammatory) increased by 53.25%, 51.68%, 83.04%, 66.50% respectively in SNP elicited plants. 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyranone (anticancerous), Deoxy-d-mannoic lactone (antibacterial) increased by 23.56% and 66.67% respectively in JA elicited plants. The present investigation gave a complete reproducible protocol for in vitro establishment of P. verticillatum. Furthermore, elicitation study on the plant helped in augmentation and authentication of neutraceutical status of the plant.
Open Access
Diversity of the family caryophyllaceae Juss. in Uttarakhand
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2016-01) Satish Chandra; Rawat, D.S.
Open Access
Assessment of morphological diversity and biochemical parameters in some selected genotypes of Trigonella foenum-graecum L.
(G.B. Pant University of Agriculture and Technology, Pantnagar (Uttarakhand), 2017-01) Nisha; Rao, P.B.
Open Access
Isolation, characterization and growth promoting attributes of plant growth promoting bacteria (PGPB) on field crops under irrigated and drought conditions
(G.B. Pant University of Agriculture and Technology, Pantnagar (Uttarakhand), 2017-01) Dinesh Chandra; Sharma, A.K.
Plant growth promoting bacteria (PGPB) containing 1-aminocyclopropane-1-carboxylate (ACC) deaminase has been reported to play an important role in alleviation of drought stress. In present study, eight ACC deaminase producing tolerant bacteria were isolated from a rainfed agriculture field of Kumaun region of Central Himalaya, Uttarakhand, India and evaluated for their efficiency in improving the growth of finger millet (Eleusine coracana (L.) Gaertn.), rice (Oryza sativa L.) and wheat (Triticum aestivum L.) under irrigated and drought conditions. These bacteria withstand a substrate metric potential of -1.0 MPa (30% PEG 8000) and therefore are considered to be drought-tolerant. The selected bacterial isolates were identified as Ochrobactrum sp., Pseudomonas sp., and Variovorax sp. by fatty acid methyl ester (FAME) analysis and 16S rRNA gene sequencing. The presence of acdS, the structural gene for ACC deaminase in selected bacterial strains was confirmed by the polymerase chain reaction (PCR). Glass house experiment showed that enhanced level of nutrients and antioxidant properties with the consortium of different bacterial strains. The consortium of Ochrobactrum anthropi DPC9 + Pseudomonas palleroniana DPB13 + Pseudomonas fluorescens DPB15 + Pseudomonas palleroniana DPB16 significantly increased the 1000 grain weight (9.3%, 28.9%), number of grains/panicle or spike (31.3%, 32.4%), grain yield (39.6%, 42.7%), and straw yield (21.1%, 26.9%) of rice and wheat, respectively as compared to their respective non-inoculated plants under organic field conditions. Variovorax paradoxus RAA3 treated plants maximally increase the total tillers/plant (40.0%, 31.7%), number of grains/spike (30.3%, 28.2%), spike length (26.4%, 25.8%), 1000 grain weight (17.4%, 22.0%), grain yield (28.2%, 47.4%), straw yield (11.2%, 26.0%) and harvest index (10.6%, 15.3%) under irrigated and rainfed conditions, respectively as compared to non-inoculated plant. Quantitative RT-PCR (qRT-PCR) based expression analysis of drought responsive genes (helicases and aquaporin) was also performed in response to V. paradoxus RAA3 inoculation in two varieties of wheat (HD 2967 and PBW 660) under irrigated and rainfed conditions The relative expression of aquaporin (TaTIP1;1) is significantly higher (7.26-fold) in the wheat variety PBW 660(drought tolerant), as compared to drought sensitive HD 2967 where expression level of aquaporin is down-regulated. The transcript level of WDH45 is down-regulated in both the varieties under irrigated conditions. In rainfed conditions, the relative mRNA level of wheat DNA helicase (WDH45) is significantly higher in HD 2967 (1.39-fold) than the PBW 660, whereas the expression pattern of aquaporin in PBW 660 is higher (2.70-fold) than the variety HD 2967 under rainfed conditions. This finding may imply PGPB having ACC deaminase activity could turn out to be effective inoculants for improving the growth of plants. Such promising strains could possibly be used under field conditions in rainfed environment where agriculture is exclusively dependent on rainfall.
Open Access
Studies on invasive alien species in Pantnagar
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2016-01) Rastogi, Jyotsna; Rawat, D.S.
Open Access
Nutraceutical potential and biochemical estimation of selected mushroom species
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2016-01) Gaur, Tanvi; Rao, P.B.
Edible mushrooms are the good source of naturally occurring biological compounds. In the present study, efforts are made to find out the nutraceutical potential of six edible mushroom species/strains viz., Agaricus bisporus, Calocybe indica, Lentinula edodes, Macrocybe giganteum MA1, Macrocybe giganteum MA2 and Pleurotus sajorcaju. All the selected species/strains possess phytochemicals viz., carbohydrates, protein, fats, phenols, flavonoids and tannins. The yield (g/10 g) with methanol in mycelia (M),freeze dried (F.D.) and oven dried (O.D.) fruit bodies varies from 0.35 ± 0.03 (P. Sajorcaju M) to 1.78 ± 0.03 (A. bisporus F.D.); and TAA (μg/mg AAE) - 12.0 ± 2.61 (L. edodes M) to 153.6 ± 2.08 (A. bisporus F.D.). The IC50 (μg/mL) ranged from 95.85 ±1.1706 (A. bisporus F.D.) to 261.7 ± 1.37 (C. indica M) DPPH; and 68.12 ± 0.58 (M. giganteum MA2 F.D.) to 114.9 ± 2.11(L. edodes M) superoxide anion scavenging activities; and EC50 (μg/mL) 95.05 ± 0.00 (M. giganteum MA2 F.D.) to 337.0 ± 1.30 (C. indica M) FCA. The reducing power activity (absorbance) varies between 0.565 ± 0.001 (P. sajor-caju F.D.) - 0.171 ± 0.001 (M. giganteum MA2 M); TPC (mg/g GAE) 3.3 ± 0.30 (M. giganteum MA1 M) to 18.0 ± 0.36 (M. giganteum MA2 F.D.); TFC (mg/g QE) 0.13 ± 0.04 (L. edodes M) to 2.03 ± 0.04 (A. bisporus F.D.); and odihydroxy phenol (mg/g CE) 0.51 ± 0.02 (A. bisporus M) to 1.10 ± 0.04 (M. Giganteum MA2 F.D.). The SOD, POD and CAT were maximum in C. indica (32.58 ± 0.01), L. edodes (6.87 ± 0.33) and C. indica (9.49 ± 0.11), respectively. The total and reducing sugars (mg/g) were maximum in C. indica (63.3 ± 1.37) and M. giganteum, MA2 (11.0± 0.85) in F.D. fruit bodies. Presence of maximum protein (36.8 ± 0.65 mg/100 g), minerals, in terms of both number and quantity; and minimum antinutrients (mg/g dw) viz., tannins (0.41 ± 0.03) and phytic acid (0.11 ± 0.01) in A. bisporus compared to others. Comparatively higher production (mm) of amylase, lipase and protease was in L. edodes (34.1 ± 1.89), P. sajor-caju (41.3 ± 1.52) and C. indica (90.0 ± 1.00), respectively. The total number of protein bands significantly decreased in both F.D. and O.D. fruit bodies compared to mycelia. The antibacterial activity was comparatively higher in C. indica against B. cereus; and M. giganteum MA2 against both E. coli and S. typhimurium. Both quercetin and rutin were absent whereas, gallic, p-hydroxy benzoic, ferulic and cinnamic acids were present in all species/strains except ferulic acid in both L. edodes and P. sajor-caju. Comparatively higher amount of total phenolics and number of bioactive compounds was in C. indica. Thus, the presence of antioxidants and nutritive components along with biologically active compounds in all the species/strains validates their use as food and pharmacology. A. bisporus, followed by M. giganteum MA2 and C. indica could be a potential source of nutraceuticals compared to other species/strains.
Open Access
Antioxidant profiling and activity in some important medicinal plant species of Uttarakhand
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2014-08) Kashyap, Shakuli; Rao, P.B.
Traditional herbal medicines form an important part of the healthcare system of India. Ayurveda is the oldest medical system in the world, provides potential leads to investigate active and therapeutically useful compounds from medicinal plant species. Considering the growing interest to ascertain the potential of medicinal plant species as important sources of natural antioxidants, the present study was conducted on 54 different plant parts sampled from 43 different selected important medicinal plant species growing in MMPG/MRDC, G.B.P.U.A. & T., Pantnagar or generally obtained in Uttarakhand state. Antioxidant profiling and activity in aqua-methanol (30:70, v/v) extracts in different plant species were performed by using three different assays namely, 2, 2-diphenyl-1-1picrylhydrazyl (DPPH) free radical scavenging, ferrous ion (Fe2+) chelating ability (FCA) and ferric reducing/antioxidant power (FRAP); non enzymatic viz., total phenolic and flavonoid content; and enzymatic antioxidants viz., superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) were analyzed in whole aerial parts (8); flowers (7); Fruits (4); seeds (2); roots (2); bark (4); and leaves (28). Comparatively higher values were found in M. pudica, O. gratissimum, T. bellirica, A. precatorius, R. serpentina, T. arjuna and Q. indica in DPPH (%); A. paniculata, C. procera, T. bellirica, S. cumini, R. serpentina, B. variegata, H. rosa-sinensis in FCA (%); E. alba, O. gratissimum, T. chebula, S. cumini, R. serpentina, T. arjuna and E. officinalis in FRAP (VCE or TE μg/mg extract); P. niruri, O. gratissimum, T. bellirica, A. precatorius, R. serpentina, T. Arjuna and B. variegata in TPC (μg GAE/mg extract); A. annua, A. annua, T. chebula, S. cumini, R. serpentina, B. variegata and B. ceiba in TFC (μg QE/mg extract); M. pudica, O. gratissimum, T. chebula, S. cumini, R. serpentina, T. arjuna and P. roxburghii in SOD (U/mg FW); B. monnieri, C. procera, T. chebula, S. cumini, A. racemosus, T. arjuna and E. officinalis in CAT (U/mg FW); and A. paniculata, O. gratissimum, T. chebula, S. cumini, R. serpentina, T. arjuna and I. coccinea in POD (U/mg FW) than other species. Thus, the above results provide scientific basis for the use of these medicinal plant species as folk remedies, promising source of natural antioxidants for the prevention and treatment of different diseases caused by oxidative stress.
Open Access
Characterization of foliar endophytic fungi from Centella asiatica (L.) Urban as a promising source of bioactive metabolites
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2015-08) Shubhpriya; Chaturvedi, Preeti
Endophytic fungi are source of novel compounds and have a great capacity to produce diverse class of plant secondary metabolites. Bioprospecting endophytes have led to exciting possibilities to explore and utilize their potential. In the present study, 13 foliar endophytic fungi were isolated from young and mature leaves of Centella asiatica (L.) Urban in summer, rainy and winter seasons. Out of 13 foliar endophytic fungi, 8 fungal isolates were morphologically identified as Colletotrichum gloeosporioides, Colletotrichum sp., Fusarium sp., Curvularia sp., Nigrospora sp., Alternaria sp., Aspergillus sp. and Fusarium equiseti and were studied further. Among all the fungal isolates, the isolation frequency of C. gloeosporioides was highest i.e. 24.14%. The qualitative screening of phytochemical constituents of ethyl acetate extract of fungal isolates revealed the presence of phenols, flavonoids, tannins, steroids, saponins, terpenoids, proteins, alkaloids, fats and carbohydrates. The different fungal isolates exhibited varied antioxidant potential amongst which C.gloeosporioides exhibited highest antioxidant activity. The maximum phenolic and flavonoid content was also found in C. gloeosporioides. A positive correlation was found between the total phenolic content (r=0.98) and total flavonoid content (r=0.86) with DPPH scavenging ability. In antimicrobial assay, all the fungal extracts showed 100% inhibition against Bacillus subtilis, 75% against Escherichia coli and 62% against Pseudomonas aeruginosa and Staphylococcus aureus. The endophytic fungal isolates were also screened for extracellular enzymes production where 62 % of endophytic fungi showed positive results for lipase, 87% for amylase, 37% for both laccase and protease and none for cellulase. Further, the endophytic fungal extracts were screened for the presence of asiaticoside by LC-MS analysis. The asiaticoside producing fungal endophyte was identified as C. gloeosporioides by ITS ¼ sequencing. C. gloeosporioides was found to produce a maximum of 62.29 μg/100mL asiaticoside in PDB medium at 15th day of incubation. In the present study it was found that production of asiaticoside by C. gloeosporioides constantly decreased from first subculture generation to seventh subculture generation. C. asiatica plants were raised in vitro for cocultivation with C. gloeosporioides. Improved growth parameters were observed in cocultivated plants compared with control plants. The cocultivated plants have higher amount of phenolics, flavonoids, DPPH free radical scavenging activity and plant defense enzymes (SOD, POX, CAT, PAL and TAL) as compared to control. The asiaticoside content was also enhanced in cocultivated plants (4.08mg/g) compared to control plants (2.63 mg/g) suggesting that C. gloeosporioides helped in value addition of cocultivated C. asiatica in vitro plants. The GC-MS analysis of methanolic extract of cocultivalted and control plants showed that cocultivated plants have higher amount of medicinally important compounds, viz. _-Farnesene, Neophytadine, _-Caryophyllene, Stigmasterol and squalene. GC-MS analysis of ethyl acetate extract of C. gloeosporioides also revealed the presence of 55 bioactive metabolites having immense antioxidant and antimicrobial activity.
Open Access
Phytotoxic efficacy and phenolic acid profiles of certain wheat and rice varieties on six weed species of Terai region, Uttarakhand
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2013-01) Rajput, Prachi; Rao, P.B.
Weeds are persistent problem for farmers due to economic losses in terms of crop yield, production costs and crop quality, which reached up to 31-61% in wheat and 36-75% in rice. It can be controlled effectively by the application of herbicides but they pose serious health and environmental threats. Allelopathy is a novel approach for minimizing the herbicide usage and to develop sustainable agriculture and also ecofreindly. The present investigation was designed to evaluate the phytotoxic potential of 15 varieties - DBW 14, JOB 668, K 9006, KAUZ/STAR, MACS 6272, PBW 550, PBW 588, RAJ 4014, RAJ 4026, RAJ 4048, RAJ 4077, UP 2382, UP 2526, UP 2565 and WH 1003 of wheat (Triticum aestivum L.) and 10 varieties - Basmati 370, Govind, Pant 11, Pant 16, Pant Sugandh 17, Pusa Sugandh 4, Pusa RH 10, Super Basmati, Tarori Basmati and VL Dhan 85 of rice (Oryza sativa L.) against six weed species - Avena ludoviciana Durieu., Celosia argentea L., Echinochloa crus-galli (L.) P. Beauv., Ischaemum rugosum Salisb., Melilotus indica L. and Phalaris minor Retz. (collected from Crop Research Centre, C.R.C.) under laboratory and green house pot experiments. The laboratory results revealed that the varieties MACS 6272, PBW 550, RAJ 4014 and RAJ 4077 of wheat and Basmati 370, Govind, PD 11 and VL Dhan 85 of rice were most inhibitory on germination, seedling growth (both length and dry weight) in different weed species. In pot experiments, both MACS 6272 and PBW 550 on E. crusgalli whereas only MACS 6272 on P. minor; and Govind on M. indica and VL Dhan 85 on both M. indica and P. minor were more suppressive in different morphological (germination, height and dry weight), physiological (chlorophyll a, b and total, carotenoid) and biochemical (proline, total protein, MDA and SOD) parameters. The reduction in different parameters in the present investigation is due to the presence of allelochemicals/secondary metabolites, chiefly phenolics. In the present study, 10 different phenolic acids were detected, identified and quantified in 15 wheat and 10 rice varieties by HPLC. The maximum phenolics (six) was identified in MACS 6272 followed by PBW 550, RAJ 4014 and RAJ 4077 (five, each) and minimum in both RAJ 4048 and WH 1003 (two each); and in rice, Govind and VL Dhan 85 (seven, each) followed by Basmati 370 (six) and lowest in both PD 16 and TB (three, each).

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