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2394 J. Med. Plants Res. DISCUSSION Genetic diversity Principal coordinates Figure 3. Two dimensional plot of Principal component analysis of nine population of Dactylorhiza using RAPD analysis. According to Hamrick and Godt (1989), there is strong relationship between geographic range and genetic diversity. There is evidence that a low level of genetic diversity is a common feature of endangered endemic plant species (Gitzendanner and Soltis, 2000). D. hatagirea is a critically endangered, self compatible and terrestrial orchid. Being terrestrial orchid, populations are small and spatially isolated. According to population genetics predict that population of such species, affected by a number of evolutionary factors including mating system, gene flow, founder effect, random genetic drift and seed dispersal (Hamrick and Godt, 1989). We compared D. hatagirea and this hypothesis with related genus Gymnadenia which is also an endangered and endemic species (Mi Yoon Chung, 2009) and our result of RAPD analysis shows that low level of genetic diversity within populations and remarkable genetic differentiation among populations in D. hatagirea. In our field survey, every year large amount of population of D. hatagirea was destroyed because of increasing mating opportunity and habitat fragmentation. Thus, habitat fragmentation under human disturbance is an important factor leading to low genetic diversity within population of D. hatagirea. So the low rate of natural recruitment observed today, together with increased habitat fragmentation and isolation of population, is seriously contributing to the low level of genetic diversity. Genetic structure The overall degree of genetic differentiation of D. hatagirea, as estimated by Gst (0.2587) is slightly higher than out crossing plants (0.23). According to an AMOVA analysis, a significantly genetically differentiation among populations was estimated (Fst = 0.259), where most genetic variation existed among populations. The genetic structure of plant populations reflects the interaction of various factors, including long–term evolutionary history of the species, genetic drift, mating system and gene flow (Hogbin and Peakall, 1999). If populations are small and isolated from one another, the genetic drift could be capable of influencing the genetic structure and increasing differentiation among populations (Ellstrand and Elam, 1993). The gene flow (Nm, 1.4703) of D. hatagirea per generation was also slightly higher than outcrossed animal pollinated species (Nm = 1.154) could be due two geographic barrier (Nubra and Shyok river) and different environmental conditions. In this study, we investigated variability among populations. Based on RAPD analysis, we revealed low genetic variation within the population and moderate genetic differentiation among the population. Therefore, every population deserves specific conservation attention, as habitat destruction and management for conservation will have greater effects on population richness. REFERENCES Avise JC, Hamrick JL (1996). Conservation genetics: Case histories from nature. New York: Chapman and Hall. Chaurasia OP, Ahmed Z, Ballabh B (2007). In: Ethnobotany and Plants of Trans-Himalaya. Satish Serial Publishing House Delhi, p. 544. Doyle JJ, Doyle JL (1987). A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull., 19: 11-15. Ellstrand NC, Elam DR (1993). Population genetic consequences of small population size: implications for plant conservation. Ann. Rev. Ecol. Syst., 24: 217e-242.
Excoffier L, Smouse PE, Quattro JM (1992). Analyses of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data. Genetics, 131: 479-491. Gitzendanner MA, Soltis PS (2000). Patterns of genetic variation in rare and endangered widespread plant congeners. Am. J. Bot., 87: 783- 792. Hamrick JL, Godt MJW (1989). Allozyme diversity in plants. In: Brown, A.H.D., Clegg, M.T., Kahler, A.L., Weir, B.S. (Eds.), Plant Population. Hogbin PM, Peakall R (1999). Evaluation of the contribution of genetic research to the management of the endangered plant Zieria prostrata. Con. Biol., 13: 514-522. Holsinger KE, Gottlieb LD (1991). Conservation of rare and endangered plants: principles and prospects. In: Falk DA, Holsinger RKE eds. Genetic and conservation of rare plants. New York: Oxford University Press, pp. 195-208. Lynch M, Milligan BG (1994). Analysis of population genetic structure within RAPD markers. Mol. Ecol., 3: 91-99. Mi YC (2009). Low level of genetic variation within populations of the four rare orchids Gymnadenia cucullata, Gymnadenia camptschatica, Amitostigma gracile, and Pogonia minor in South Korea: indication of genetic drift and implications for conservation. Plant Syst. Evol., 281: 65-76. Milligan BG, Leebens-Mack J, Strand AE (1994). Conservation genetics: beyond the maintenance of marker diversity. Mol. Ecol., 12: 844–855. Nei M (1973). Analysis of gene diversity in subdivided populations. PNAS, 70: 3321-3323. Nei M (1978). Estimation of average heterozygosity and genetic distance from a small number of individual. Genetis, 89: 583-590. Warghat et al. 2395 Peakall R, Smouse PE (2001). GenAlEx V5 Genetic Analysis in Excel. Population genetic software for teaching and research. Australian National University, Canberra, Australia, (http://www.anu.edu.au/BoZo/ GenAlEx/). Prevost A, Wilkinson MJ (1999). A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor. Appl. Genet., 98: 107-112. Rohlf FJ (1992). NTSYS-PC: Numerical taxonomy and multivariate analysis system version 2.02. State University of New York (Stony Brook, New York). Williams JGK, Kubelik AR, Livak KL, Rafalski JA, Tingey SV (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl. Acids Res., 18: 6531-6535. Wright S (1951). The genetical structure of populations. Ann. Eugen., 15: 323-354. Yeh FC, Yang RC, Boyle T (1999). POPGENE. Microsoft Windowsbased freeware for population genetic analysis. Release 1.31. Edmonton: University of Alberta. Zhao WG, Zhang JQ, Wangi YH, Chen TT, Yin Y, Huang YP, Pan Y, Yang Y (2006). Analysis of genetic diversity in wild populations of mulberry from western part of Northeast China determined by ISSR markers. J. Genet. Mol. Biol., 7: 196-203.
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Journal of Medicinal Plants Researc
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Editors Prof. Akah Peter Achunike E
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Electronic submission of manuscript
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Fees and Charges: Authors are requi
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Table of Contents: Volume 6 Number
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Figure 1. Chuanxiong Rhizoma (http:
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Table 1. Identification of main pea
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Table 3. Pharmacokinetic parameters
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active component study, many invest
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and 1185 kg ha -1 in the first site
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Table 1. Recommendation for use fer
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Sadanandan AK, Hamza S (1996c). Stu
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and it has been traced to South Ame
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Table 3. Results of phytochemical a
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(a) (b) (c) Figure 1. (a) Normal ce
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emedies in the south of Brazil. J.
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known fatty acids and terpenoids we
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Table 1. Renal function tests of ra
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Al-Radahe et al. 2271 Figure 3.nnnn
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namely disruption of the vascular e
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Zayachkivska OS, Konturek SJ, Drozd
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β-carotene were purchased from Sig
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Figure 1. Effect of ethanol concent
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Table 3. Climatic and soil factors
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content of polysaccharides. Phospha
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Serge et al. 2285 Table 1. Relative
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Percentage inhibition % of inhibiti
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Table 1. Result of phytochemical sc
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Table 3. Result of thin layer chrom
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Table 2. MIC of a compound 1 from c
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Zirihi et al. 2301 Figure 2. Termin
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Figure 5. T. catappa Linn. (Combret
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Figure 8. A. fumigatus: Aspect of c
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Figure 12. Sensitivity of A. fumiga
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Noormi et al. 2311 Table 1. Callus
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Table 2. Contd. NoC=No callus. 4.0
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Figure 2. Contd. at 2.0 mg/L yellow
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Table 1. Biochemical parameters in
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Figure 3. Bcl-xL reactions in inter
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ACKNOWLEDGMENTS The authors would l
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Figure 1. The first page of the boo
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Figure 3. The most widely included
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Figure 4. Lithographic print “St.
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Table 1. List of plants included in
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Table 1. Contd. Nedelcheva 2333 Cin
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14 7 1 15 1 35 17 16 Imported plant
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Academy of Science Publishing House
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et al., 1996; van Wyk and Gericke,
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2466 J. Med. Plants Res. AOAC (2000
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2486 J. Med. Plants Res. components
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2490 J. Med. Plants Res. Inhibition
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UPCOMING CONFERENCES 15th European
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