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Brochure for Book Chapter, We are happy to inform you that we are going to publish an edited book with ISBN, under the proposed title of "Possibilities of Organic Farming in India” from ASTRAL, INTERNATIONAL PVT. LTD., NEW DELHI, ISBN number will be allotted soon. The, book contains the latest information on organic farming. The book is writing in a very, simple language, which is easy to understand. This book will prove to be a milestone for, the undergraduate and postgraduate students of Agriculture, teachers, and it is also of, great use to the progressive farmers who are intended to take up organic farming in, agriculture field. This book is a significant step on the part of academicians/researchers, because it recognizes the potentials of ideas and strategies to combat the developing, challenges in technology, economics, agriculture practices, business/economic, weeds,, diseases and insect pest management in organic farming. So, we invite contributors, working in different fields to publish his/her scholastic and original article in our edited, book., Important Guidelines for the Authors, ➢ The word limit of full chapters with abstract of 150-200 words and 4-5 keywords, is maximum 5000 words and 08 pages., ➢ Full paper must be written in 'British English', typed using 'Times New Roman', (Font Size 12) and in 'MS-Word', APA 6th Format must be followed for, references. Submit the softcopy of manuscript electronically as an attachment in, the, following, email, ([email protected], or, [email protected]), ➢ It should be free from plagiarism., ➢ Tables, Charts or Graphs (if any) must be put in the main body of the Article, itself. The Editorial Board will review the whole article/chapter. They are the sole, authority for giving consent of publication of the article., ➢ All views and opinions expressed in the book are the sole responsibility of the, author concerned. Neither the editor nor publisher can in anyway, be held, responsible for them., Related Areas:, 1. Importance and scope of organic farming., 2. Production technologies of field crops., 3. Production technologies of horticultural crops., 4. Preparation and uses of organic inputs (Compost, Vermi-compost, Bio-fertilizers,, FYM, Green manuring etc.) for organic farming., 5. Crop residues management of different crops.

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6. Nutrients management in organic farming (Field and Horticultural crops)., 7. Weed management in organic farming (Field and Horticultural crops)., 8. Diseases management in organic farming (Field and Horticultural crops)., 9. Insect pest management in organic farming (Field and Horticultural crops)., 10. Secondary Agriculture (Value Addition) in organic products., 11. Marketing and distributions of organic products., 12. Organic foods and certification., 13. Economics of organic crops., 14. Integrated organic farming system., *Articles/chapters not only constraint to the above topics but also the other topics, directly related to organic farming and its consequences will be willingly accepted., Review charge of the book chapter and publication fee accordingly rupees 1500/(one thousand five hundreds) only including author copy price. Co-author copy, charges rupees 1200/- (one thousand two hundred) will be extra., Last date of submission of book chapters and payment receipt 20-11-2021, Intimation of acceptance 30-11-2021, , Account details:, Account holder name: Suman Kumari, Account no. 6564000100038566, IFSC: PUNB0656400, Bank: Panjab National Bank, Bulandshahr, , Branch: Awas Vikas-I, DM Road,, , *Fifth character of IFSC is Zero (0), All communication should be maintained to the following E- mails, [email protected],, inquiries., Editors-, , [email protected], , Feel, , free, , for, , further

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Demo Chapter, PHYTOCHEMICAL CHARACTERIZATION AND ANTIOXIDANT, PROFILING OF MOSSES FROM PITHORAGARH, KUMAUN, HIMALAYA, A…….. R……1, N…..2 and P……. C………1,*, 1, , Department of Biological Sciences, G.B. Pant University of Agriculture & Technology,, Pantnagar (Uttarakhand) India, 2, , Department of Botany, Government P.G. College, Sector-1 Panchkula (Haryana), *E-mail: ………[email protected], , Abstract, Bryophytes, a group of smaller and apparently simpler stature plants are the oldest known plants, on land. The three major groups of bryophytes namely mosses, liverworts, and hornworts, comprise the earliest lineages of land plants derived from green algal ancestors. Bryophytes,, despite their miniature size, occupy a unique position in an ecosystem. Widespread regeneration, capacity and rich reserve of phytochemicals are the major reasons behind their successful, colonization and cosmopolitan distribution. Present study is an attempt to characterize the, phytoconstituents of three mosses collected from Pithoragarh (1925-1960m asl) viz., Leucodon, secundus (Harv.) Mitt., Plagiomnium integrum (Bosch & Sande Lac.) T.J. Kop. and Thuidium, cymbifolium (Dozy & Molk.) Dozy & Molk. P. integrum and T. cymbifolium inhabit the forest, floor while L. secundus is epiphytic in nature. Biochemical characterization was studied in terms, of total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activity via 2, 2–, Diphenyl Picryl Hydrazyl (DPPH) free radical scavenging activity and total antioxidant assay, (TAA). DPPH radical scavenging activity and TFC was found highest (79.82±0.65%;, 94.025±0.94 mg QE g-1) in L. secundus. Maximum TAA (88.57±0.70) and TPC (38.50±0.69 mg, GAE g-1) was reported in T. cymbifolium while minimum in L. Secundus (12.55±0.07; 6.23±0.17, mg GAE g-1)., Key words: Bryophytes, mosses, total flavonoid content, total phenolic content, DPPH activity,, antioxidant activity

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integrum, E80°12’52.8’’, (Bosch & Sande Lac.), T.J. Kop., Thuidium cymbifolium 1925, N29°32’11.7’’, Terrestrial, 3.2, Dozy & Molk.) Dozy, E80°12’52.8’’, & Molk, The plant samples were identified by consulting the available taxonomic literature viz., Chopra,, 1975 and Gangulee, 1969. Voucher specimens of the mosses were deposited in the herbarium of, G.B. Pant University of Agriculture & Technology, Pantnagar., 2.2 Preparation of Plant Extract, The plant thalli with rhizoids were extensively washed with water and then spread on the paper, sheet and dried in the open air at room temperature. The dried plant material was pulverized in, an electric grinder and stored in polythene bags, then dipped in absolute methanol in the ratio, 1:10 (1g in 10 ml) and shaken in an orbital shaker for 5 days at 120 rpm at 35°C. The extract was, filtered and centrifuged at 15000 rpm at 4°C. The supernatant was dried at room temperature to, obtain the dried crude extracts. The stocks were kept in the refrigerator for further, experimentation. The yield of the sample (Table 1) was calculated as percentage, according to, the formula mentioned below:, Yield (%) = Total crude extract (g) / dried powder (g) X 100, 2.3. Biochemical Characterization, •, , Phytochemical Screening, , Preliminary qualitative phytochemical analysis was done to identify the phytoconstituents, present in the methanol extract of the three mosses following the standard methods, (Table 2)., •, , 2, 2– Diphenyl Picryl Hydrazyl (DPPH) Free Radical Scavenging Activity:, Antioxidant activity of methanol extracts was evaluated in terms of 2, 2’-diphenyl-1-, , picrylhydrazyl (DPPH) radical scavenging ability following Blois (1958). Various concentrations, of extracts (20, 40, 60, 80, 100µg ml-1) were mixed with 1ml of freshly prepared DPPH soln., (0.1mM of DPPH in methanol). The mixture was incubated at room temperature for 30 min. and, the absorbance was measured at 517 nm in a UV-Vis spectrophotometer (Thermo Scientific).

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BHT was used as a positive control. DPPH radical scavenging activity was calculated by the, following equation:, % DPPH radical scavenging activity = [1-(As/Ac)] × 100, (Where, As is the absorbance of the sample and Ac is the absorbance of the control at 517nm)., The experiment was conducted in triplicates., , • Total Antioxidant Activity (TAA), Phospho-molybdenum method was used for the evaluation of the total antioxidant activity. The assay, is based on the reduction of Mo (VI) to Mo (V) by the extract and subsequent formation of a, green phosphate/Mo (V) complex at acidic pH (Prieto et al., 1999). Different concentrations (20,, 40, 60, 80 and 100 µg/ml) of the extract was taken in the test tube containing 3ml of reagent soln., (0.6 M sulphuric acid, 28 mM sodium phosphate and 4 mM ammonium molybdate) and, incubated at 95 °C for 90 min. The absorbance of each sample, was measured at 695 nm by, spectrophotometer, after cooling at room temperature, and compared against blank and expressed, as ascorbic acid equivalents., , • Total Phenolic Content (TPC), The total phenolic content was determined by the method given by Singleton and Rossi (1965)., 0.5ml of the plant extract of different concentrations was transferred into a test tube and mixed, with 4.5 ml of dist. water followed by addition of 0.2ml of FCR (Folin Ciocalteau Reagent)., After 3 min, 7% of sodium carbonate was added and finally, the volume was made up to 5ml, with the dist. water. The mixture was allowed to stand for 1 hour and the absorbance of the, sample was taken at 765 nm. A standard curve was established using various concentrations of, gallic acid (1mg/ml) in 80% methanol and results were expressed as gallic acid equivalents, (GAE µg)/ mg extract., •, , Total Flavonoid Content (TFC), , Estimation of the total flavonoids in the plant extracts was carried out using the method of, Ordonez et al. (2006). Extract samples were evaluated at a final concentration of 100µg ml-1., Total flavonoid content was calculated as quercetin equivalents (mg/ml) using the following, equation based on the calibration curve: y = 0.020 x, where x was the absorbance and y was the

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quercetin equivalent (mg/ml). Quercetin (1 mg/ml) stock was prepared in methanol and its, different concentrations were used to obtain the calibration curve. TFC values were expressed as, µg quercetin equivalent per mg extract)., , • Statistical analysis, All the experiments were carried out using five replicates each in different independent, experiments. Results were expressed as mean ± standard error (SE) The data were statistically, analysed by two-way ANOVA using SPSS software (version 17.0)., 3. Result and Discussion, •, , Phytochemical Screening, , The results of preliminary phytochemical screening of methanol extract of the mosses are given, in Table 2., Table 2. Qualitative phytochemical screening in the three moss species, Phytochemicals, L. secundus, Carbohydrates (Molisch test), +, Proteins (Xanthoproteic test), +, Fats (Stain test), Glycosides (Keller- Killani test), Flavonoids (Sulphuric acid test), +, Terpenoids (Salkowski test), +, Alkaloids (Mayer’s reagent test,, Wagner’s reagent test ), Tannins (FeCl3 test), Phenols (Lead acetate test), +, Saponins (Foam test), +, Quinones (HCl test), (+) indicates presence, (-) indicates absence, , P. integrum, +, +, +, +, +, +, -, , T. cymbifolium, +, +, +, +, +, +, -, , In the present study, methanol was used as the solvent based on the previous literature available, from the lab (Negi and Chaturvedi, 2016; Negi et al., 2018). Methanol extracts of all the three, mosses showed presence of carbohydrates, proteins, flavonoids, terpenoids, phenols as well as, saponins. The methanolic extracts of the mosses, however, tested negative for fats, glycosides,

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alkaloids, tannins and quinines (Table 2). This may be because either these compounds are, absent in the mosses or the solvent used was not able to solubilise these phytochemicals., •, , 2, 2– Diphenyl Picryl Hydrazyl (DPPH) Free Radical Scavenging Activity:, , All the tested extracts showed dose dependent DPPH radical scavenging activity (Fig. 1). DPPH, radical scavenging activity was highest in L. Secundus (79.82± 0.65) followed by T., cymbifolium(68.58± 1.07). P. integrum showed minimum DPPH radical scavenging activity, (56.63± 0.28)., , • Total Antioxidant Activity (TAA), Total antioxidant activity (TAA) was however, significantly higher in T. cymbifolium (88.57±, 0.70) than the other two mosses (Fig.2). Both antioxidant activity (in terms of DPPH radical, scavenging activity) and TAA is plant specific depending on the individual plants., , • Total Phenolic Content (TPC), Total phenolic content (TPC) was evaluated with the help of gallic acid calibration curve, obtained colorimetrically and expressed as µg gallic acid equivalents (GAE)/mg extract., Significantly high TPC was reported in T. cymbifolium (38.50 ± 0.69) compared to the other, extracts (Fig. 3)., •, , Total Flavonoid Content (TFC), , Contrary to the results of TPC, total flavonoid content (TFC) was significantly higher in L., secundus (94.02± 0.94) and P. integrum (84.25± 1.04) and comparatively low TFC was found in, T. cymbifolium (28.2 ± 0.29). McCleary et al. (1960) reported antibiotic activity of P. cuspidatum, and several other bryophytes. The phytochemical studies on Thuidium tamariscellum showed the, presence of significant level of terpenoids in the moss. High antioxidant property shown by the, plant is reported to be mainly due to the presence of considerable amount of terpenoids, (Mohandas and Kumarswamy, 2018)., In the present study also, the total flavonoid content was highest in epiphyte, L. secundus and, least in T. cymbifolium (Fig. 4). This was correlated with DPPH radical scavenging power of the, respective extracts. Collection data given in Table 1 clearly showed that L. secundus is epiphytic

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in nature while T. cymbifolium and P. integrum were terrestrial. Epiphytic habitat of L. secundus, exposed it to comparatively unusual growth conditions. To cope with these conditions, there was, more accumulation of flavonoids and other compounds that gave survival benefit to the plant., Highest total flavonoid content of epiphytic bryophytes was also reported by Wang et al. (2017)., It could be due to the reason that epiphytic species were more exposed in the air and experienced, complex and varied environments. This required them to synthesize flavonoids and other, protective compounds. These compounds thus increased their antioxidant capacity providing, them biochemical defence in the exposed habitat. Hence, not only the genetic makeup of the, species but also its ecology play a major role in directing the production and accumulation of, different phytochemicals as part of survival strategy., , DPPH radical scavenging activity (%), , L. secundus, , P. integrum, , T. cymbifolium, , 90, 80, , b, , 70, 60, , c, , c, , c, , c, , c, , b, , b, , b, , b, , a, , a, , a, , a, , a, , 20, , 40, , 60, , 80, , 10, , 50, 40, 30, 20, 10, 0, Concentrations (µg/ml), , Fig. 1. DPPH free radical scavenging activity (%) in methanol extract of different, bryophytes ( expressed as mean of five different conc. of DPPH values with S.E.)

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Total antioxidant activity (µg AAE/mg), , L. secundus, , P. integrum, , T. cymbifolium, , 100, , b, , 90, c, , 80, 70, c, , 60, 50, , b, , 40, 30, , b, , 20, 10, , a, , a, , a, , a, , a, , a, , b, , b, , a, , a, , 0, , 20, , 40, , 60, , 80, , 100, , Concentrations (µg/ml), , Fig. 2. Total antioxidant activity in methanol extract of different bryophytes (expressed, as mean of five different conc. of TAA values with S.E.), 45, b, , Total phenolic content (µg GAE/mg), , 40, 35, 30, 25, 20, 15, 10, a, , a, , 5, 0, L. secundus, , P. integrum, Bryophyte extracts, , T. cymbifolium

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Fig. 3. Total phenolic content in methanol extract of different bryophytes (expressed as, mean of five different conc. of TAA values with S.E.), , Total flavonoid content (µg QE/mg), , 100, , c, , 90, , b, , 80, 70, 60, 50, , 40, a, , 30, 20, 10, 0, L. secundus, , P. integrum, Bryophyte extracts, , T. cymbifolium, , Fig. 4. Total flavonoid content in methanol extract of different bryophytes (expressed as, mean of five different conc. of TFC values with S.E.), , 4. Conclusion, Needless to say, bryophytes lack any specific food value. However, it is their rich chemical, constitution which makes them an interesting group in the plant kingdom. It is this biochemical, makeup which is responsible for their desiccation and disease tolerant nature equipping them, with unique survival mechanisms. Mosses are a rich storehouse of bioactive compounds viz.,, phenolics and flavonoids. These phytoconstituents are responsible for the antioxidant properties,, either synergistically or individually. Further, the type and concentration of phytoconstituents, and their resulting biological activities are plant and habitat-specific. Surely, these humble plants, provide a rich reservoir of chemicals that have immense medicinal and economic values., Acknowledgements