Chemical analysis, biological activity determination and DNA profiling of different accessions of Curcuma longa L. collected from Uttarakhand
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Date
2014-08
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G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand)
Abstract
Polyphenolics, known as nutritional secondary metabolities, plays an important role to protect the tissues by reducing the oxidative stress and enhance the nutritional value of food material. The total curcumin content, essential elements, phytochemical screening and antioxidant activity of 50 samples of Curcuma longa L. rhizome from various niches of Uttrakhand (Kumaun and Gharwal region) were analyzed. The highest curcumin content was found in the sample collected form Pithoragarh (37.93±0.12mg/g) and lowest content was in Patwadanger (9.29±0.51mg/g) collection. The Fe, Cu, Zn and Mn contents was found in the range of 20.89 to 3462.58, 1.11 to 198.03, 3.54 to 211.75 and 5.56 to 1108.80 mg/kg respectively. These contents were maximum in sample collected from Dholia pata (Bageshwar), Nanakmatta (Udhamsingh Nagar), Pantnagar (Udhamsingh Nagar) and Champawat and minimum in the sample collected from Kathgodam (Nainital), Harsil (Uttarkashi, Garhwal), Bhimtal (Nainital) and Dharchula (Pithoragarh). The P, Ca, Mg, K and N content was found in the range of 0.04 to 0.99, 0.19 to 1.42, 0.09 to 0.85, 0.57 to 2.99 and 9.52 to 35.28% respectively. The P and Ca contents were maximum in the sample collected from Devprayag (Chamoli), Khatima (Udhamsingh nagar), Mg and K in Chamoli and N in Roorkee (Haridwar). This content was found minimum in sample collected from Barswar (Pauri Garhwal), Pithoragarh, Ramnagar (Udhamsingh Nagar) Kanda (Bageshwar) and Dwarahat (Almora). The amount of total phenols varied from 6.71±1.29 to 51.49±1.41mg/g. The maximum phenolic content was found in Pithoragarh collection and minimum in Khatima (Udhamsingh nagar) collection. The total flavonoid contents varied from 6.34±1.81 to 28.96±2.47 mg/g with maximum flavonoid in the sample collected from Kapkot (Bageshwar) and minimum in Kanda (Bageshwar) collection. Orthodihydroxy phenol (ODP) varied from 0.69±0.10 to 8.11±0.11 mg catechol equivalent/g. The maximum ODP was found in the sample collected from Chamoli and minimum in Chakrata (Dehradun). The content of ascorbic acid varied from 0.122 ± 0.008 to 2.61 ± 0.0071mg/g. The total sugar, reducing and non-reducing content in all the fifty collections varied from 56.25±0.77 -- 13.75±0.72, 11.2±0.39 -- 1.2±0.10 and 49.67±0.84 -- 8.58±1.24 mg/g respectively. The Munsiyari (Pithoraghar) collection showed maximum amount of total sugar and non-reducing sugar while Nanakmatta (Udhamsingh Nagar), Deharadun and Pantnagar (Udhamsingh Nagar) showed the minimum amount. The total Protein content in different samples ranged from 4.87±0.08 to 24.52±0.11 mg/g with maximum in Roorkee (Haridwar) collection and minimum in Chakrata (Dehradun) collection. The other samples showed the amounts in between of maximum and minimum and all the samples exhibited in vitro – antioxidant activity in a dose dependent manner. The total antioxidant capacity and FRAP was found in the range of 27.44±2.5 to 94.78±3.4 mg/g and 6.99±.39 to 54.84±.53 μmole FeSO4 equvalient/g respectivily. The methanolic extracts also showed DPPH radical scavenging activity, Hydroxy radical scavenging activity, superoxide radical scavenging activity and nitric oxide radical scavenging activity in dose dependent manner but significantly less compared to the standards (EDTA, ascorbic acid and qurecitin). The IC50 values of these activities were varied from 55.93±0.25 to 275.41±0.53, 39.57±.42 to 77.68±.63, 43.76±0.53 to77.12±0.59 and 39.02±0.46 to 76.15±0.39μg/ml respectively. Out of fifty samples two samples selected from Ukhimath and Haldwani, showed good anti-inflammatory, antinociceptive and antipyretic activities in dose dependent manner. None of the extract exhibited toxicity at 400, 600 and 800 mg/kg concentration. The RAPD technique was performed to detect genetic diversity in the twenty turmeric sample. Ten RAPD primers yielded 84 alleles, averaging 8.4 alleles per locus varying from seven to ten. Polymorphism index contents values ranging from 0.24 to 0.77 with the mean of 0.43. The observed heterozygosity (Ho) and gene diversity (Nei’s) for individual loci varied from 0.158 to 0.672 and 0.154 to 0.564, respectively. The Shannon’s informative index (I) of loci varied from 0.137 to 0.846 with the mean of 0.425 per locus. Fixation index (Fis), was also calculated, which ranged from 0.125 to 0.600 with an average of 0.377. A genetic relationship among accessions were analyzed by cluster analysis using unweighted pair group method with arithmetic mean (UPGMA), the average-linkage method with the similarity matrix as input data and classified all twenty turmeric accessions into two major groups comprising six clusters. From result, the samples collected from Lohaghat and Patanpatni have the same parents and sample collected from Harsil and Munsiyari and from kanda and Champawat are closely related accessions. The wide geographical and climatic distribution is indicative of the fact that there exists a tremendous genetic diversity in Curcuma longa which needs to be identified and catalogued for scientific and breeding programmes for their commercial usages.
Description
Thesis-PhD
Keywords
chemical analysis, biological activity, determination, DNA, accessory chromosomes, turmeric, Curcuma longa, Uttarakhand, metabolites