DEPARTMENT OF BIOTECHNOLOGY COLLEGE OF AGRICULTURE JUNAGADH AGRICULTURAL UNIVERSITY JUNAGADH
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Date
2013-07
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jau,junagadh
Abstract
Fruits of black jamun have remarkable antidiabetic properties. Phytochemicals found in fruit parts (pulp, seed, seed coat, kernel) are varied with differencial degree of antidiabetic activity. So, present experiment is planned with objective to determine phytochemicals, antioxidant and antidiabetic activities from the extract of various solvent fractions (Petroleum ether, Ethyl acetate, Methanol, Water) derived from fruit parts. Total six black jamun landraces is categorized on the basis of fruit size and other morphological characters. Large fruit (BJLR 1) contained higher amount of moisture, total fatty matter, ash content, ascorbic acid, anthocynine, starch content, free amino acid. While, smaller fruits (BJLR 6) have higher amount of crude fibre, total protein and total phenol in their fruit pulp and seed. Quality parameters were observed higher in pulp compare to seed except crude fibre, total protein and total phenol which were recorded higher in seed.
The extraction of fruit parts (pulp, seed, seed coat, kernel) was carried out in different solvents. Methanol and water extracts showed more extractive yield as compared to petroleum ether and ethyl acetate extracts. The qualitative assay of phytochemicals like alkaloids, amino acid, anthraquinon, flavanoids, glycosides, phenol, saponins, steroids, tannins, triterpenoids, antioxidant and antidiabetic activities were carried out from different solvent fraction of fruit parts. Most of the phytochemicals were highly present in methanolic fraction including antioxidant and antidiabetic activities.
Among the landraces, the seeds of BJLR 6 were exhibited highest (95.4% inhibition, IC50 4.2 μg.ml-1) antioxidant activity. Comparatively, seed contained higher antioxidant activity particularly because of its kernel fraction. The highest antidiabetic activity (98.2% inhibition of PPA) was recorded in smallest fruits (BJLR 6), particularly metabolic fraction of seed parts followed by BJLR 5 (93.8%). Quantification of major phytochemicals like phenol, flavanoids, glycosides, alkaloids
and terpenoids were carried out from methanoloic fraction of fruit parts of six landraces. The seeds of all landraces contained higher amount of phenol, flavanoids, alkaloids and glycosides while pulp tissues exhibited high terpenoids. The seeds of BJLR 6 (smaller fruits) landraces elevated antidiabetic activity because of high of phenol, flavanoids, glycosides and alkaloids than the other landraces.
Total 7 phenolics (gallic acid, catechin, chlorogenic acid, caffeic acid, ferullic acid, ellagic acid, quarcetine) were identified from the methanolic extract of fruit parts of black jamun landraces using HPLC. The seed hold high amount of all seven phenolics followed by kernel, in which seed of BJLR 6 (best landraces with respect to antidiabetic activity) have high amount of gallic acid, chlorogenic acid, caffeic acid, ferullic acid, ellagic acid. The seed and kernel of BJLR 6 contained quarcetine which may responsible for antidiabetic activity. Total 12 compounds were identified from the pulp and seed tissue (methanolic fraction) of BJLR 6 landraces in GC-MS. However, total 22 compounds were identified from the seed coat, and 10 compounds from the kernel. Major compound were found to be N'-isopropylureidoacetic acid (37.63%) followed by Cyclopropylcarbinol (30.71 %) in pulp; 2, butynoic acid (36.40 %) followed by Spiro [4.4]nonane-1,6-dione (13.67 %) in seed; Bicyclo ethylamine (51.1%) followed by Hydroxyurea (15.9 %) in seed coat; Pyrazole 5- carboxylic acid (36.88 %), Furan carboxaldehyde (34.31 %) in kernel of BJLR 6 landraces.
The landraces is also subjected to DNA profiling with aim to identify landraces which possess high antidiabetic properties. Out of 21, eighteen ISSR primers amplified total 144 bands with average 8 bands per primer. Out of total 144 bands, 50 were monomorphic and 94 were polymorphic with average 5.2 bands per primer. Cluster analysis clearly outgrouped large fruit size landraces BJLR 1, which have low antidiabetic activity, from other five landraces and shared 42 % similarity. However, BJLR 6 (smallest fruit size) landrace having highest antidiabetic activity in their fruit parts distinguished from other four landraces (moderate fruit size) and shared 53 % similarity. Total 32 unique bands were recorded by 18 UBC primers to discriminate 6 landraces of black jamun. Maximum 12 unique bands were found by 6 ISSR primers for identification BJLR 6 landraces, which have high antidiabetic activity in their fruit parts (pulp, seed, seed coat and kernel). Primer UBC 834 were found to be more informative as it provided 4 unique bands (275, 498, 595, 711 bp) for identification of most diverse BJLR 6 landraces and one unique (1451 bp) for discrimination of BJLR 1 which was poor in antidiabetic activity.
Key words: Black Jamun, Fruit parts, Antidiabetic activity, Antioxidant activity, Phytochemcals, HPLC, GC-MS, DNA profiling.
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biotechnology