Thirty seven genotypes ofMorus albaL. from the trans-Himalaya with white, pink and black fruit we... more Thirty seven genotypes ofMorus albaL. from the trans-Himalaya with white, pink and black fruit were
studied tofind out variability and relationship of fruit color and sampling location with total antioxidant
capacity, total polyphenol content (TPC), totalflavonoid content (TFC) and total anthocyanin content
(TAC) in fruit. The ferric reducing antioxidant potential (FRAP) ranged from 34.2 to 46.2mmol Fe(II)/g DW.
TPC ranged from 1.8 to 8.3 mg gallic acid equivalent (GAE)/g DW. The TFC ranged from 0.4 to 0.9 mg
quercetin equivalent/g DW. Two way analysis of variance (ANOVA) revealed that fruit color has no significant relationship with TPC and TAC. However, fruit color showed inconsistent effect on antioxidant
capacity as determined by different methods. Sampling location has significant effect on antioxidant
capacity, TPC, TFC and TAC. Interaction between fruit color and sampling location showed significant
effect on antioxidant capacity and TAC. The study therefore showed that source of genotypes from
different sampling location is more important as compare to fruit color in determining antioxidant capacity, TPC, TFC and TAC inM. albafruit from the trans-Himalaya.
Journal of Systematics and Evolution, Aug 19, 2014
Genome scan was done with 439 dominant marker loci to identify outlier loci in three populations ... more Genome scan was done with 439 dominant marker loci to identify outlier loci in three populations ofMorus
albafrom the trans‐Himalayan region. Employingfinite and hierarchical island population genetic models, we
detected 30 (6.85%) loci under selection, of which 3.64% were under directional selection and 3.19% under balancing
selection. The hierarchical island model was efficient in detecting signatures of natural selection with low occurrence
of false positives. Data showed significant spatial genetic structure (SGS), especially in population from Nubra valley
with significantly high SGS. Bayesian approach revealed high genetic differentiation and inbreeding in Nubra
population. Indus and Suru valleys populations showed comparatively low SGS, genetic differentiation, and
inbreeding. The study showed thatM. albafalls in the range of mixed mating and outcrossing species. However, the
population of Nubra valley showed selfing with high SGS. Significant SGS strengthens the presence of natural
selection inM. albanatural populations.
Sequence-related amplified polymorphism markers were used to assess the genetic structure in thre... more Sequence-related amplified polymorphism markers were used to assess the genetic structure in three natural populations of Morus alba from trans-Himalaya. Multilocation sampling was conducted across 14 collection sites. The overall genetic diversity estimates were high: percentage polymorphic loci 89.66%, Nei’s gene diversity 0.2286, and Shannon’s information index 0.2175. At a regional level, partitioning of variability assessed using analysis of molecular variance (AMOVA), revealed 80% variation within and 20% among collection sites. Pattern appeared in STRUCTURE, BARRIER, and AMOVA, clearly demonstrating gene flow between the Indus and Suru populations and a geographic barrier between the Indus-Suru and Nubra populations, which effectively hinders gene flow. The results showed significant genetic differentiation, population structure, high to restricted gene flow, and high genetic diversity. The assumption that samples collected from the three valleys represent three different populations does not hold true. The fragmentation present in trans-Himalaya was more natural and less anthropogenic.
BMC Complementary and Alternative Medicine, Oct 7, 2013
Background:
Himalayan plants are widely used in traditional system of medicine both as prophylac... more Background:
Himalayan plants are widely used in traditional system of medicine both as prophylactics and
therapeutics for high altitude maladies. Our aim was to evaluate the antioxidant capacities and bioactive compounds of
methanol and n-hexane extracts of the phytococktail comprising of sea buckthorn (Hippophae rhamnoides), apricot
(Prunus armeniaca) and roseroot (Rhodiola imbricata) from trans-Himalaya.
Methods:
The 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium
salt (ABTS) and nitric oxide (NO) radical scavenging capacities and lipid peroxidation inhibition (LPI) property of the
extracts were determined. Total antioxidant power was determined by ferric reducing/antioxidant power (FRAP) assay.
Total polyphenol, flavonoid, flavonol, proanthocyanidin and carotenoid were also estimated for both extracts. We have
identified and quantified the phyto-chemotypes present in the methanol and n-hexane extracts by hyphenated gas
chromatography/mass spectrometry (GC/MS) technique.
Results:
Antioxidant capacity assays using DPPH, ABTS, NO, LPI and FRAP exhibited analogous results where the
phytococktail showed high antioxidant action. The phytococktail was also found to possess high quantity of total
polyphenol, flavonoid, flavonol and carotenoid. A significant and linear correlation was found between the antioxidant
capacities and bioactive principles. A total of 32 phyto-chemotypes were identified from these extracts by GC/MS
chemometric fingerprinting. Major phyto-chemotypes identified by GC/MS were glycosides, phenylpropanoids and
derivatives, terpenoids, alkaloids, phytosterols, fatty acids and esters, alkaloids and derivatives, organic acid esters and
aromatic ethers with positive biological and pharmacological actions.
Conclusion:
The phytococktail extracts were found to contain considerable amount of diverse bioactive compounds
with high antioxidant capacities. The presence of hydrophilic and lipophilic antioxidants in the phytococktail could
have contributed to the higher antioxidant values. Hence, the phytococktail could be used as natural source of
antioxidants to ameliorate disorders associated with oxidative stress.
The aim of this study was to evaluate thein vitro antioxidant potential of hydro-ethanolic extrac... more The aim of this study was to evaluate thein vitro antioxidant potential of hydro-ethanolic extract of a novel
phytococktail comprising of sea buckthorn, apricot, andRhodiola(SAR) from trans-Himalaya. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) activity of the extract increased in a dose-dependent manner (upto 0.1 mg/mL), and was found
to be about 38% of that of ascorbic acid at 0.1 mg/mL. The hydro-ethanolic extract of SAR also scavenged the ABTS
.+
radical generated by ABTS/potassium persulfate (PPS) system and was found to be about 62% of that of ascorbic
acid at 0.1 mg/ mL. The total antioxidant power of the extract was determined by ferric reducing antioxidant power
(FRAP) assay. Total phenolic content was found to be 1.28016×10
−3
mol gallic acid equivalent (GAE)/g extract. Total
flavonoid and flavonol contents were estimated to be 2.5970×10
−4
mol and 4.87×10
−4
mol quercetin equivalent/g
extract, respectively. The hydro-ethanolic extract of this phytococktail indicated presence of essential phytoconstituents
of polyphenols, flavonoids, flavonols, and ascorbic acid, which contributed significantly to its antioxidant capacity. The
combination of the 3 plants may well support their use in traditional medicine to combat oxidative stress and high-altitude
sickness.
Immature seeds were cultured on 10 different media or germination. Maximum germination was achiev... more Immature seeds were cultured on 10 different media or germination. Maximum germination was achieved on Lindeman orchid medium (37.12%) within 17 days o culturing. Protocorms with lea primordia were cultured on BM-2 and 7 different modifications o Murashige and Skoog (MS) medium with various hormone combinations [0–3 mg/L indole butyric acid (IBA) and 0–3 mg/L kinetin (Kin)] or plantlet regeneration and mass multiplication. Maximum number o shoots (18.12 ± 0.3), highest shoot length (17.80 cm ± 2.16), maximum root number (8.25 ± 0.69), and highest root length (8.02 cm ± 1.45) were ound on MS medium with 3 mg/L IBA and 1 mg/L Kin. Plantlets with 2–3 shoots were transerred to different potting mixtures or acclimatization to field conditions and urther multiplication. One hundred percent survival was obtained in C-8 potting mixture consisting o cocopeat + vermiculite + perlite (1:1:1), which produced 75 shoots (25 plantlets) afer 1 month o transplantation in the greenhouse. Te current study describes or the first time a rapid in vitro protocorm development and mass multiplication protocol or
Dactylorhiza hatagirea
(D.Don) Soo that holds robust potential or large-scale propagation and metabolite production.
Analysis of molecular variance (AMOVA) of ISSR analysis revealed a moderate level of ... more Analysis of molecular variance (AMOVA) of ISSR analysis revealed a moderate level of genetic differentiation among populations (60% of the total variance components, P < 0.001), with gene differentiation
coefficients (Gst = 0.258, Fst = 0.135) and moderate gene flow (Nm = 1.432). Both Principal coordinate’s
analysis (PCoA) and Neighbor-Joining (NJ) cluster analysis supported the grouping of all 96 sample sizes of
nine locations into two collection groups. The present level and pattern of genetic diversity and structure
of Dactylorhiza hatagirea were assumed to result largely from its habitat fragmentation, its unique biological traits and evolutionary history. The genetic structure could be attributed to an earlier period of more
pronounced gene flow when the species had a more continuous distribution. A Mantel test revealed no significant positive correlation between genetic distances and geographic distances (R
2
= 0.0165, P = 0.290).
The aim of the study was to evaluate the antioxidant and free radical scavenging properties, tota... more The aim of the study was to evaluate the antioxidant and free radical scavenging properties, total phenolic contents and antibacterial activities of the methanol extract of
Mentha longifolia
Linn. Hudson. growing in the high altitude cold desert region of Ladakh, Jammu and Kashmir, India. The antioxidant activities of the extracts were investigated with four different methods such as 2,2’-diphenyl-1-picrylhydrazyl radical (DPPH), 2,2’-azinobis (3-ethylbenzothi-azoline-6-sulfonic acid) radical (ABTS), Reducing power assay and Ferric reducing antioxidant property (FRAP).
M . longifolia
exhibited the strongest activity as ABTS scavenger compared to DPPH and FRAP. All the assays carried out found that the plant has antioxidant potential. The total polyphenol content (TPC) and total flavonoid content of the extract was determined as 107.208 ± 34.2 mg GAE/g dry wt. and 42.47 ± 7.97 mg QE/g dry wt. respectively where 39.61% of TPC were flavonoids. A high correlation was found between the DPPH and ABTS free radical scavenging potency (R
2
= 0.869). The study has validated the medicinal potential of
Mentha longifolia
Linn. Hudson and revealed that the consumption of the plant could be beneficial as it possesses the desirable quality of antioxidant activity
Journal of Medicinal Plants Research, Mar 30, 2012
Random amplified polymorphic DNA (RAPD) analysis was used to characterize the genetic diversity a... more Random amplified polymorphic DNA (RAPD) analysis was used to characterize the genetic diversity and population genetic structure within and among nine natural populations of
Dactylorhiza hatagirea,
a critically endangered or rare terrestrial medicinal orchid in cold desert of Ladakh region. Out of the 177 bands generated from twenty random primers, 174 were polymorphic. The genetic diversity of
D. hatagirea
that was revealed by observed number of alleles (N
a
), expected number of alleles (Ne), Nei’s diversity index (H), Shannon’s diversity ind
ex (I), amplificated loci, polymorphic loci and the percentage of polymorphic loci (PPL). Pair-wise population genetic distances ranged from 0.05 to 0.23. Analysis of molecular variance (AMOVA) revealed that 57% RAPD of variability was partitioned among population.
The Principal coordinate’s analysis (PCoA) and UPGMA supported the grouping of all 96 accessions of
nine populations into four cluster groups. However, a moderate genetic differentiation among population was detected based on different measures (
Nei’s genetic diversity analysis: Gst = 0.2538;
AMOVA analysis: Fst = 0.254)
The morphometric study was conducted during 2009 to 2010. About 28 morphological characters werem... more The morphometric study was conducted during 2009 to 2010. About 28 morphological characters weremeasured under 13 natural populations of
Dactylorhiza hatagirea.
Geographic variation in morphologyreflects phenotypic responses to environmental gradients and evolutionary history of populations andspecies. At points, beside its broad geographic range (Nubra, Suru and Indus valley) characterization of
Dactylorhiza
phenotype is normally accomplished by use of morphological descriptors, hence as a firststep, phenotype collection and its morphometric analysis was assessed. However, plant height, leaflength, lowermost leaf length, length of second leaf from base and mean length from lowest bract to thetop of inflorescence are presented to account for the remarkable variation in morphological characters.Tirith population showed more values of this trait while Skurru showed less value. From this, it isconcluded that Tirith showed great morphometric variation as compared to other population.Multivariate morphometric techniques, principal component analysis (PCA), multidimensional scaling(MDS) and cluster analysis were used to determine whether these populations can be reliablymorphologically similar or dissimilar. The first two principal components encompass more than 75%variation among population. The results of PCA and MDS analysis were comparable to the clusteranalysis, which shows considerable phenotypic variation in morphological and horticultural traits thatcan be utilized in its genetic improvement. To support this study, further constructive information wereprovided on the status of the populations of
D. hatagirea
which may increase the conservation value ofthis site and resolve the suitable areas with taxonomic and nomenclatural controversies.
Journal of Medicinal Plants Research, Jan 23, 2012
Rhodiola imbricata
Edgew. (Crassulaceae), well acknowledged medicinal plant is widely distribut... more Rhodiola imbricata
Edgew. (Crassulaceae), well acknowledged medicinal plant is widely distributed intrans-Himalayan region of India. It has multiple uses in cuisine, forage, health care and ornamentalworth. In the present investigation, 70 wildly grown
Rhodiola
genotypes, collected from three differentsampling sites in trans-Himalayan region of Ladakh were analyzed using 40 DNA-based markers (20random amplified polymorphic DNAs (RAPDs) and 20 inter simple sequence repeats (ISSRs). RAPDanalysis yielded 134 fragments, of which 130 were polymorphic, with an average of 6.5 polymorphicfragments per primer. Of the 20 ISSR primer screened, only 8 were amplified in the presentinvestigation. These amplified primers produced 59 bands, of which 57 were polymorphic with anaverage of 7.12 polymorphic fragments per primer. RAPD markers were found more efficient withregards to polymorphism detection, as they detected 98.28% as compared to 97.55% for ISSR markers.Clustering of genotypes within groups was remained more or less same when RAPD, ISSR and RAPD +ISSR derived dendrograms were compared. The results of principal coordinates analysis (PCA) analysiswere corresponding to the dendrogram analysis. These analyses allowed us to identify the groupscorresponding to the three
Rhodiola
collection sites. Analysis of molecular variance (AMOVA) showedthat genetic variation within the populations was found maximum than among populations in all thethree cases. It may because of high level of differentiation within populations due to geographical andgenetic isolation of populations in a harsh mountainous environment. With reference to themanagement of
Rhodiola
, the high genetic separation of population indicates the requisite ofconserving the extreme possible number of populations from diverse parts of trans-Himalayas.
Thirty seven genotypes ofMorus albaL. from the trans-Himalaya with white, pink and black fruit we... more Thirty seven genotypes ofMorus albaL. from the trans-Himalaya with white, pink and black fruit were
studied tofind out variability and relationship of fruit color and sampling location with total antioxidant
capacity, total polyphenol content (TPC), totalflavonoid content (TFC) and total anthocyanin content
(TAC) in fruit. The ferric reducing antioxidant potential (FRAP) ranged from 34.2 to 46.2mmol Fe(II)/g DW.
TPC ranged from 1.8 to 8.3 mg gallic acid equivalent (GAE)/g DW. The TFC ranged from 0.4 to 0.9 mg
quercetin equivalent/g DW. Two way analysis of variance (ANOVA) revealed that fruit color has no significant relationship with TPC and TAC. However, fruit color showed inconsistent effect on antioxidant
capacity as determined by different methods. Sampling location has significant effect on antioxidant
capacity, TPC, TFC and TAC. Interaction between fruit color and sampling location showed significant
effect on antioxidant capacity and TAC. The study therefore showed that source of genotypes from
different sampling location is more important as compare to fruit color in determining antioxidant capacity, TPC, TFC and TAC inM. albafruit from the trans-Himalaya.
Journal of Systematics and Evolution, Aug 19, 2014
Genome scan was done with 439 dominant marker loci to identify outlier loci in three populations ... more Genome scan was done with 439 dominant marker loci to identify outlier loci in three populations ofMorus
albafrom the trans‐Himalayan region. Employingfinite and hierarchical island population genetic models, we
detected 30 (6.85%) loci under selection, of which 3.64% were under directional selection and 3.19% under balancing
selection. The hierarchical island model was efficient in detecting signatures of natural selection with low occurrence
of false positives. Data showed significant spatial genetic structure (SGS), especially in population from Nubra valley
with significantly high SGS. Bayesian approach revealed high genetic differentiation and inbreeding in Nubra
population. Indus and Suru valleys populations showed comparatively low SGS, genetic differentiation, and
inbreeding. The study showed thatM. albafalls in the range of mixed mating and outcrossing species. However, the
population of Nubra valley showed selfing with high SGS. Significant SGS strengthens the presence of natural
selection inM. albanatural populations.
Sequence-related amplified polymorphism markers were used to assess the genetic structure in thre... more Sequence-related amplified polymorphism markers were used to assess the genetic structure in three natural populations of Morus alba from trans-Himalaya. Multilocation sampling was conducted across 14 collection sites. The overall genetic diversity estimates were high: percentage polymorphic loci 89.66%, Nei’s gene diversity 0.2286, and Shannon’s information index 0.2175. At a regional level, partitioning of variability assessed using analysis of molecular variance (AMOVA), revealed 80% variation within and 20% among collection sites. Pattern appeared in STRUCTURE, BARRIER, and AMOVA, clearly demonstrating gene flow between the Indus and Suru populations and a geographic barrier between the Indus-Suru and Nubra populations, which effectively hinders gene flow. The results showed significant genetic differentiation, population structure, high to restricted gene flow, and high genetic diversity. The assumption that samples collected from the three valleys represent three different populations does not hold true. The fragmentation present in trans-Himalaya was more natural and less anthropogenic.
BMC Complementary and Alternative Medicine, Oct 7, 2013
Background:
Himalayan plants are widely used in traditional system of medicine both as prophylac... more Background:
Himalayan plants are widely used in traditional system of medicine both as prophylactics and
therapeutics for high altitude maladies. Our aim was to evaluate the antioxidant capacities and bioactive compounds of
methanol and n-hexane extracts of the phytococktail comprising of sea buckthorn (Hippophae rhamnoides), apricot
(Prunus armeniaca) and roseroot (Rhodiola imbricata) from trans-Himalaya.
Methods:
The 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium
salt (ABTS) and nitric oxide (NO) radical scavenging capacities and lipid peroxidation inhibition (LPI) property of the
extracts were determined. Total antioxidant power was determined by ferric reducing/antioxidant power (FRAP) assay.
Total polyphenol, flavonoid, flavonol, proanthocyanidin and carotenoid were also estimated for both extracts. We have
identified and quantified the phyto-chemotypes present in the methanol and n-hexane extracts by hyphenated gas
chromatography/mass spectrometry (GC/MS) technique.
Results:
Antioxidant capacity assays using DPPH, ABTS, NO, LPI and FRAP exhibited analogous results where the
phytococktail showed high antioxidant action. The phytococktail was also found to possess high quantity of total
polyphenol, flavonoid, flavonol and carotenoid. A significant and linear correlation was found between the antioxidant
capacities and bioactive principles. A total of 32 phyto-chemotypes were identified from these extracts by GC/MS
chemometric fingerprinting. Major phyto-chemotypes identified by GC/MS were glycosides, phenylpropanoids and
derivatives, terpenoids, alkaloids, phytosterols, fatty acids and esters, alkaloids and derivatives, organic acid esters and
aromatic ethers with positive biological and pharmacological actions.
Conclusion:
The phytococktail extracts were found to contain considerable amount of diverse bioactive compounds
with high antioxidant capacities. The presence of hydrophilic and lipophilic antioxidants in the phytococktail could
have contributed to the higher antioxidant values. Hence, the phytococktail could be used as natural source of
antioxidants to ameliorate disorders associated with oxidative stress.
The aim of this study was to evaluate thein vitro antioxidant potential of hydro-ethanolic extrac... more The aim of this study was to evaluate thein vitro antioxidant potential of hydro-ethanolic extract of a novel
phytococktail comprising of sea buckthorn, apricot, andRhodiola(SAR) from trans-Himalaya. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) activity of the extract increased in a dose-dependent manner (upto 0.1 mg/mL), and was found
to be about 38% of that of ascorbic acid at 0.1 mg/mL. The hydro-ethanolic extract of SAR also scavenged the ABTS
.+
radical generated by ABTS/potassium persulfate (PPS) system and was found to be about 62% of that of ascorbic
acid at 0.1 mg/ mL. The total antioxidant power of the extract was determined by ferric reducing antioxidant power
(FRAP) assay. Total phenolic content was found to be 1.28016×10
−3
mol gallic acid equivalent (GAE)/g extract. Total
flavonoid and flavonol contents were estimated to be 2.5970×10
−4
mol and 4.87×10
−4
mol quercetin equivalent/g
extract, respectively. The hydro-ethanolic extract of this phytococktail indicated presence of essential phytoconstituents
of polyphenols, flavonoids, flavonols, and ascorbic acid, which contributed significantly to its antioxidant capacity. The
combination of the 3 plants may well support their use in traditional medicine to combat oxidative stress and high-altitude
sickness.
Immature seeds were cultured on 10 different media or germination. Maximum germination was achiev... more Immature seeds were cultured on 10 different media or germination. Maximum germination was achieved on Lindeman orchid medium (37.12%) within 17 days o culturing. Protocorms with lea primordia were cultured on BM-2 and 7 different modifications o Murashige and Skoog (MS) medium with various hormone combinations [0–3 mg/L indole butyric acid (IBA) and 0–3 mg/L kinetin (Kin)] or plantlet regeneration and mass multiplication. Maximum number o shoots (18.12 ± 0.3), highest shoot length (17.80 cm ± 2.16), maximum root number (8.25 ± 0.69), and highest root length (8.02 cm ± 1.45) were ound on MS medium with 3 mg/L IBA and 1 mg/L Kin. Plantlets with 2–3 shoots were transerred to different potting mixtures or acclimatization to field conditions and urther multiplication. One hundred percent survival was obtained in C-8 potting mixture consisting o cocopeat + vermiculite + perlite (1:1:1), which produced 75 shoots (25 plantlets) afer 1 month o transplantation in the greenhouse. Te current study describes or the first time a rapid in vitro protocorm development and mass multiplication protocol or
Dactylorhiza hatagirea
(D.Don) Soo that holds robust potential or large-scale propagation and metabolite production.
Analysis of molecular variance (AMOVA) of ISSR analysis revealed a moderate level of ... more Analysis of molecular variance (AMOVA) of ISSR analysis revealed a moderate level of genetic differentiation among populations (60% of the total variance components, P < 0.001), with gene differentiation
coefficients (Gst = 0.258, Fst = 0.135) and moderate gene flow (Nm = 1.432). Both Principal coordinate’s
analysis (PCoA) and Neighbor-Joining (NJ) cluster analysis supported the grouping of all 96 sample sizes of
nine locations into two collection groups. The present level and pattern of genetic diversity and structure
of Dactylorhiza hatagirea were assumed to result largely from its habitat fragmentation, its unique biological traits and evolutionary history. The genetic structure could be attributed to an earlier period of more
pronounced gene flow when the species had a more continuous distribution. A Mantel test revealed no significant positive correlation between genetic distances and geographic distances (R
2
= 0.0165, P = 0.290).
The aim of the study was to evaluate the antioxidant and free radical scavenging properties, tota... more The aim of the study was to evaluate the antioxidant and free radical scavenging properties, total phenolic contents and antibacterial activities of the methanol extract of
Mentha longifolia
Linn. Hudson. growing in the high altitude cold desert region of Ladakh, Jammu and Kashmir, India. The antioxidant activities of the extracts were investigated with four different methods such as 2,2’-diphenyl-1-picrylhydrazyl radical (DPPH), 2,2’-azinobis (3-ethylbenzothi-azoline-6-sulfonic acid) radical (ABTS), Reducing power assay and Ferric reducing antioxidant property (FRAP).
M . longifolia
exhibited the strongest activity as ABTS scavenger compared to DPPH and FRAP. All the assays carried out found that the plant has antioxidant potential. The total polyphenol content (TPC) and total flavonoid content of the extract was determined as 107.208 ± 34.2 mg GAE/g dry wt. and 42.47 ± 7.97 mg QE/g dry wt. respectively where 39.61% of TPC were flavonoids. A high correlation was found between the DPPH and ABTS free radical scavenging potency (R
2
= 0.869). The study has validated the medicinal potential of
Mentha longifolia
Linn. Hudson and revealed that the consumption of the plant could be beneficial as it possesses the desirable quality of antioxidant activity
Journal of Medicinal Plants Research, Mar 30, 2012
Random amplified polymorphic DNA (RAPD) analysis was used to characterize the genetic diversity a... more Random amplified polymorphic DNA (RAPD) analysis was used to characterize the genetic diversity and population genetic structure within and among nine natural populations of
Dactylorhiza hatagirea,
a critically endangered or rare terrestrial medicinal orchid in cold desert of Ladakh region. Out of the 177 bands generated from twenty random primers, 174 were polymorphic. The genetic diversity of
D. hatagirea
that was revealed by observed number of alleles (N
a
), expected number of alleles (Ne), Nei’s diversity index (H), Shannon’s diversity ind
ex (I), amplificated loci, polymorphic loci and the percentage of polymorphic loci (PPL). Pair-wise population genetic distances ranged from 0.05 to 0.23. Analysis of molecular variance (AMOVA) revealed that 57% RAPD of variability was partitioned among population.
The Principal coordinate’s analysis (PCoA) and UPGMA supported the grouping of all 96 accessions of
nine populations into four cluster groups. However, a moderate genetic differentiation among population was detected based on different measures (
Nei’s genetic diversity analysis: Gst = 0.2538;
AMOVA analysis: Fst = 0.254)
The morphometric study was conducted during 2009 to 2010. About 28 morphological characters werem... more The morphometric study was conducted during 2009 to 2010. About 28 morphological characters weremeasured under 13 natural populations of
Dactylorhiza hatagirea.
Geographic variation in morphologyreflects phenotypic responses to environmental gradients and evolutionary history of populations andspecies. At points, beside its broad geographic range (Nubra, Suru and Indus valley) characterization of
Dactylorhiza
phenotype is normally accomplished by use of morphological descriptors, hence as a firststep, phenotype collection and its morphometric analysis was assessed. However, plant height, leaflength, lowermost leaf length, length of second leaf from base and mean length from lowest bract to thetop of inflorescence are presented to account for the remarkable variation in morphological characters.Tirith population showed more values of this trait while Skurru showed less value. From this, it isconcluded that Tirith showed great morphometric variation as compared to other population.Multivariate morphometric techniques, principal component analysis (PCA), multidimensional scaling(MDS) and cluster analysis were used to determine whether these populations can be reliablymorphologically similar or dissimilar. The first two principal components encompass more than 75%variation among population. The results of PCA and MDS analysis were comparable to the clusteranalysis, which shows considerable phenotypic variation in morphological and horticultural traits thatcan be utilized in its genetic improvement. To support this study, further constructive information wereprovided on the status of the populations of
D. hatagirea
which may increase the conservation value ofthis site and resolve the suitable areas with taxonomic and nomenclatural controversies.
Journal of Medicinal Plants Research, Jan 23, 2012
Rhodiola imbricata
Edgew. (Crassulaceae), well acknowledged medicinal plant is widely distribut... more Rhodiola imbricata
Edgew. (Crassulaceae), well acknowledged medicinal plant is widely distributed intrans-Himalayan region of India. It has multiple uses in cuisine, forage, health care and ornamentalworth. In the present investigation, 70 wildly grown
Rhodiola
genotypes, collected from three differentsampling sites in trans-Himalayan region of Ladakh were analyzed using 40 DNA-based markers (20random amplified polymorphic DNAs (RAPDs) and 20 inter simple sequence repeats (ISSRs). RAPDanalysis yielded 134 fragments, of which 130 were polymorphic, with an average of 6.5 polymorphicfragments per primer. Of the 20 ISSR primer screened, only 8 were amplified in the presentinvestigation. These amplified primers produced 59 bands, of which 57 were polymorphic with anaverage of 7.12 polymorphic fragments per primer. RAPD markers were found more efficient withregards to polymorphism detection, as they detected 98.28% as compared to 97.55% for ISSR markers.Clustering of genotypes within groups was remained more or less same when RAPD, ISSR and RAPD +ISSR derived dendrograms were compared. The results of principal coordinates analysis (PCA) analysiswere corresponding to the dendrogram analysis. These analyses allowed us to identify the groupscorresponding to the three
Rhodiola
collection sites. Analysis of molecular variance (AMOVA) showedthat genetic variation within the populations was found maximum than among populations in all thethree cases. It may because of high level of differentiation within populations due to geographical andgenetic isolation of populations in a harsh mountainous environment. With reference to themanagement of
Rhodiola
, the high genetic separation of population indicates the requisite ofconserving the extreme possible number of populations from diverse parts of trans-Himalayas.
Uploads
studied tofind out variability and relationship of fruit color and sampling location with total antioxidant
capacity, total polyphenol content (TPC), totalflavonoid content (TFC) and total anthocyanin content
(TAC) in fruit. The ferric reducing antioxidant potential (FRAP) ranged from 34.2 to 46.2mmol Fe(II)/g DW.
TPC ranged from 1.8 to 8.3 mg gallic acid equivalent (GAE)/g DW. The TFC ranged from 0.4 to 0.9 mg
quercetin equivalent/g DW. Two way analysis of variance (ANOVA) revealed that fruit color has no significant relationship with TPC and TAC. However, fruit color showed inconsistent effect on antioxidant
capacity as determined by different methods. Sampling location has significant effect on antioxidant
capacity, TPC, TFC and TAC. Interaction between fruit color and sampling location showed significant
effect on antioxidant capacity and TAC. The study therefore showed that source of genotypes from
different sampling location is more important as compare to fruit color in determining antioxidant capacity, TPC, TFC and TAC inM. albafruit from the trans-Himalaya.
albafrom the trans‐Himalayan region. Employingfinite and hierarchical island population genetic models, we
detected 30 (6.85%) loci under selection, of which 3.64% were under directional selection and 3.19% under balancing
selection. The hierarchical island model was efficient in detecting signatures of natural selection with low occurrence
of false positives. Data showed significant spatial genetic structure (SGS), especially in population from Nubra valley
with significantly high SGS. Bayesian approach revealed high genetic differentiation and inbreeding in Nubra
population. Indus and Suru valleys populations showed comparatively low SGS, genetic differentiation, and
inbreeding. The study showed thatM. albafalls in the range of mixed mating and outcrossing species. However, the
population of Nubra valley showed selfing with high SGS. Significant SGS strengthens the presence of natural
selection inM. albanatural populations.
Himalayan plants are widely used in traditional system of medicine both as prophylactics and
therapeutics for high altitude maladies. Our aim was to evaluate the antioxidant capacities and bioactive compounds of
methanol and n-hexane extracts of the phytococktail comprising of sea buckthorn (Hippophae rhamnoides), apricot
(Prunus armeniaca) and roseroot (Rhodiola imbricata) from trans-Himalaya.
Methods:
The 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium
salt (ABTS) and nitric oxide (NO) radical scavenging capacities and lipid peroxidation inhibition (LPI) property of the
extracts were determined. Total antioxidant power was determined by ferric reducing/antioxidant power (FRAP) assay.
Total polyphenol, flavonoid, flavonol, proanthocyanidin and carotenoid were also estimated for both extracts. We have
identified and quantified the phyto-chemotypes present in the methanol and n-hexane extracts by hyphenated gas
chromatography/mass spectrometry (GC/MS) technique.
Results:
Antioxidant capacity assays using DPPH, ABTS, NO, LPI and FRAP exhibited analogous results where the
phytococktail showed high antioxidant action. The phytococktail was also found to possess high quantity of total
polyphenol, flavonoid, flavonol and carotenoid. A significant and linear correlation was found between the antioxidant
capacities and bioactive principles. A total of 32 phyto-chemotypes were identified from these extracts by GC/MS
chemometric fingerprinting. Major phyto-chemotypes identified by GC/MS were glycosides, phenylpropanoids and
derivatives, terpenoids, alkaloids, phytosterols, fatty acids and esters, alkaloids and derivatives, organic acid esters and
aromatic ethers with positive biological and pharmacological actions.
Conclusion:
The phytococktail extracts were found to contain considerable amount of diverse bioactive compounds
with high antioxidant capacities. The presence of hydrophilic and lipophilic antioxidants in the phytococktail could
have contributed to the higher antioxidant values. Hence, the phytococktail could be used as natural source of
antioxidants to ameliorate disorders associated with oxidative stress.
phytococktail comprising of sea buckthorn, apricot, andRhodiola(SAR) from trans-Himalaya. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) activity of the extract increased in a dose-dependent manner (upto 0.1 mg/mL), and was found
to be about 38% of that of ascorbic acid at 0.1 mg/mL. The hydro-ethanolic extract of SAR also scavenged the ABTS
.+
radical generated by ABTS/potassium persulfate (PPS) system and was found to be about 62% of that of ascorbic
acid at 0.1 mg/ mL. The total antioxidant power of the extract was determined by ferric reducing antioxidant power
(FRAP) assay. Total phenolic content was found to be 1.28016×10
−3
mol gallic acid equivalent (GAE)/g extract. Total
flavonoid and flavonol contents were estimated to be 2.5970×10
−4
mol and 4.87×10
−4
mol quercetin equivalent/g
extract, respectively. The hydro-ethanolic extract of this phytococktail indicated presence of essential phytoconstituents
of polyphenols, flavonoids, flavonols, and ascorbic acid, which contributed significantly to its antioxidant capacity. The
combination of the 3 plants may well support their use in traditional medicine to combat oxidative stress and high-altitude
sickness.
Dactylorhiza hatagirea
(D.Don) Soo that holds robust potential or large-scale propagation and metabolite production.
coefficients (Gst = 0.258, Fst = 0.135) and moderate gene flow (Nm = 1.432). Both Principal coordinate’s
analysis (PCoA) and Neighbor-Joining (NJ) cluster analysis supported the grouping of all 96 sample sizes of
nine locations into two collection groups. The present level and pattern of genetic diversity and structure
of Dactylorhiza hatagirea were assumed to result largely from its habitat fragmentation, its unique biological traits and evolutionary history. The genetic structure could be attributed to an earlier period of more
pronounced gene flow when the species had a more continuous distribution. A Mantel test revealed no significant positive correlation between genetic distances and geographic distances (R
2
= 0.0165, P = 0.290).
Mentha longifolia
Linn. Hudson. growing in the high altitude cold desert region of Ladakh, Jammu and Kashmir, India. The antioxidant activities of the extracts were investigated with four different methods such as 2,2’-diphenyl-1-picrylhydrazyl radical (DPPH), 2,2’-azinobis (3-ethylbenzothi-azoline-6-sulfonic acid) radical (ABTS), Reducing power assay and Ferric reducing antioxidant property (FRAP).
M . longifolia
exhibited the strongest activity as ABTS scavenger compared to DPPH and FRAP. All the assays carried out found that the plant has antioxidant potential. The total polyphenol content (TPC) and total flavonoid content of the extract was determined as 107.208 ± 34.2 mg GAE/g dry wt. and 42.47 ± 7.97 mg QE/g dry wt. respectively where 39.61% of TPC were flavonoids. A high correlation was found between the DPPH and ABTS free radical scavenging potency (R
2
= 0.869). The study has validated the medicinal potential of
Mentha longifolia
Linn. Hudson and revealed that the consumption of the plant could be beneficial as it possesses the desirable quality of antioxidant activity
Dactylorhiza hatagirea,
a critically endangered or rare terrestrial medicinal orchid in cold desert of Ladakh region. Out of the 177 bands generated from twenty random primers, 174 were polymorphic. The genetic diversity of
D. hatagirea
that was revealed by observed number of alleles (N
a
), expected number of alleles (Ne), Nei’s diversity index (H), Shannon’s diversity ind
ex (I), amplificated loci, polymorphic loci and the percentage of polymorphic loci (PPL). Pair-wise population genetic distances ranged from 0.05 to 0.23. Analysis of molecular variance (AMOVA) revealed that 57% RAPD of variability was partitioned among population.
The Principal coordinate’s analysis (PCoA) and UPGMA supported the grouping of all 96 accessions of
nine populations into four cluster groups. However, a moderate genetic differentiation among population was detected based on different measures (
Nei’s genetic diversity analysis: Gst = 0.2538;
AMOVA analysis: Fst = 0.254)
Dactylorhiza hatagirea.
Geographic variation in morphologyreflects phenotypic responses to environmental gradients and evolutionary history of populations andspecies. At points, beside its broad geographic range (Nubra, Suru and Indus valley) characterization of
Dactylorhiza
phenotype is normally accomplished by use of morphological descriptors, hence as a firststep, phenotype collection and its morphometric analysis was assessed. However, plant height, leaflength, lowermost leaf length, length of second leaf from base and mean length from lowest bract to thetop of inflorescence are presented to account for the remarkable variation in morphological characters.Tirith population showed more values of this trait while Skurru showed less value. From this, it isconcluded that Tirith showed great morphometric variation as compared to other population.Multivariate morphometric techniques, principal component analysis (PCA), multidimensional scaling(MDS) and cluster analysis were used to determine whether these populations can be reliablymorphologically similar or dissimilar. The first two principal components encompass more than 75%variation among population. The results of PCA and MDS analysis were comparable to the clusteranalysis, which shows considerable phenotypic variation in morphological and horticultural traits thatcan be utilized in its genetic improvement. To support this study, further constructive information wereprovided on the status of the populations of
D. hatagirea
which may increase the conservation value ofthis site and resolve the suitable areas with taxonomic and nomenclatural controversies.
Edgew. (Crassulaceae), well acknowledged medicinal plant is widely distributed intrans-Himalayan region of India. It has multiple uses in cuisine, forage, health care and ornamentalworth. In the present investigation, 70 wildly grown
Rhodiola
genotypes, collected from three differentsampling sites in trans-Himalayan region of Ladakh were analyzed using 40 DNA-based markers (20random amplified polymorphic DNAs (RAPDs) and 20 inter simple sequence repeats (ISSRs). RAPDanalysis yielded 134 fragments, of which 130 were polymorphic, with an average of 6.5 polymorphicfragments per primer. Of the 20 ISSR primer screened, only 8 were amplified in the presentinvestigation. These amplified primers produced 59 bands, of which 57 were polymorphic with anaverage of 7.12 polymorphic fragments per primer. RAPD markers were found more efficient withregards to polymorphism detection, as they detected 98.28% as compared to 97.55% for ISSR markers.Clustering of genotypes within groups was remained more or less same when RAPD, ISSR and RAPD +ISSR derived dendrograms were compared. The results of principal coordinates analysis (PCA) analysiswere corresponding to the dendrogram analysis. These analyses allowed us to identify the groupscorresponding to the three
Rhodiola
collection sites. Analysis of molecular variance (AMOVA) showedthat genetic variation within the populations was found maximum than among populations in all thethree cases. It may because of high level of differentiation within populations due to geographical andgenetic isolation of populations in a harsh mountainous environment. With reference to themanagement of
Rhodiola
, the high genetic separation of population indicates the requisite ofconserving the extreme possible number of populations from diverse parts of trans-Himalayas.
studied tofind out variability and relationship of fruit color and sampling location with total antioxidant
capacity, total polyphenol content (TPC), totalflavonoid content (TFC) and total anthocyanin content
(TAC) in fruit. The ferric reducing antioxidant potential (FRAP) ranged from 34.2 to 46.2mmol Fe(II)/g DW.
TPC ranged from 1.8 to 8.3 mg gallic acid equivalent (GAE)/g DW. The TFC ranged from 0.4 to 0.9 mg
quercetin equivalent/g DW. Two way analysis of variance (ANOVA) revealed that fruit color has no significant relationship with TPC and TAC. However, fruit color showed inconsistent effect on antioxidant
capacity as determined by different methods. Sampling location has significant effect on antioxidant
capacity, TPC, TFC and TAC. Interaction between fruit color and sampling location showed significant
effect on antioxidant capacity and TAC. The study therefore showed that source of genotypes from
different sampling location is more important as compare to fruit color in determining antioxidant capacity, TPC, TFC and TAC inM. albafruit from the trans-Himalaya.
albafrom the trans‐Himalayan region. Employingfinite and hierarchical island population genetic models, we
detected 30 (6.85%) loci under selection, of which 3.64% were under directional selection and 3.19% under balancing
selection. The hierarchical island model was efficient in detecting signatures of natural selection with low occurrence
of false positives. Data showed significant spatial genetic structure (SGS), especially in population from Nubra valley
with significantly high SGS. Bayesian approach revealed high genetic differentiation and inbreeding in Nubra
population. Indus and Suru valleys populations showed comparatively low SGS, genetic differentiation, and
inbreeding. The study showed thatM. albafalls in the range of mixed mating and outcrossing species. However, the
population of Nubra valley showed selfing with high SGS. Significant SGS strengthens the presence of natural
selection inM. albanatural populations.
Himalayan plants are widely used in traditional system of medicine both as prophylactics and
therapeutics for high altitude maladies. Our aim was to evaluate the antioxidant capacities and bioactive compounds of
methanol and n-hexane extracts of the phytococktail comprising of sea buckthorn (Hippophae rhamnoides), apricot
(Prunus armeniaca) and roseroot (Rhodiola imbricata) from trans-Himalaya.
Methods:
The 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium
salt (ABTS) and nitric oxide (NO) radical scavenging capacities and lipid peroxidation inhibition (LPI) property of the
extracts were determined. Total antioxidant power was determined by ferric reducing/antioxidant power (FRAP) assay.
Total polyphenol, flavonoid, flavonol, proanthocyanidin and carotenoid were also estimated for both extracts. We have
identified and quantified the phyto-chemotypes present in the methanol and n-hexane extracts by hyphenated gas
chromatography/mass spectrometry (GC/MS) technique.
Results:
Antioxidant capacity assays using DPPH, ABTS, NO, LPI and FRAP exhibited analogous results where the
phytococktail showed high antioxidant action. The phytococktail was also found to possess high quantity of total
polyphenol, flavonoid, flavonol and carotenoid. A significant and linear correlation was found between the antioxidant
capacities and bioactive principles. A total of 32 phyto-chemotypes were identified from these extracts by GC/MS
chemometric fingerprinting. Major phyto-chemotypes identified by GC/MS were glycosides, phenylpropanoids and
derivatives, terpenoids, alkaloids, phytosterols, fatty acids and esters, alkaloids and derivatives, organic acid esters and
aromatic ethers with positive biological and pharmacological actions.
Conclusion:
The phytococktail extracts were found to contain considerable amount of diverse bioactive compounds
with high antioxidant capacities. The presence of hydrophilic and lipophilic antioxidants in the phytococktail could
have contributed to the higher antioxidant values. Hence, the phytococktail could be used as natural source of
antioxidants to ameliorate disorders associated with oxidative stress.
phytococktail comprising of sea buckthorn, apricot, andRhodiola(SAR) from trans-Himalaya. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) activity of the extract increased in a dose-dependent manner (upto 0.1 mg/mL), and was found
to be about 38% of that of ascorbic acid at 0.1 mg/mL. The hydro-ethanolic extract of SAR also scavenged the ABTS
.+
radical generated by ABTS/potassium persulfate (PPS) system and was found to be about 62% of that of ascorbic
acid at 0.1 mg/ mL. The total antioxidant power of the extract was determined by ferric reducing antioxidant power
(FRAP) assay. Total phenolic content was found to be 1.28016×10
−3
mol gallic acid equivalent (GAE)/g extract. Total
flavonoid and flavonol contents were estimated to be 2.5970×10
−4
mol and 4.87×10
−4
mol quercetin equivalent/g
extract, respectively. The hydro-ethanolic extract of this phytococktail indicated presence of essential phytoconstituents
of polyphenols, flavonoids, flavonols, and ascorbic acid, which contributed significantly to its antioxidant capacity. The
combination of the 3 plants may well support their use in traditional medicine to combat oxidative stress and high-altitude
sickness.
Dactylorhiza hatagirea
(D.Don) Soo that holds robust potential or large-scale propagation and metabolite production.
coefficients (Gst = 0.258, Fst = 0.135) and moderate gene flow (Nm = 1.432). Both Principal coordinate’s
analysis (PCoA) and Neighbor-Joining (NJ) cluster analysis supported the grouping of all 96 sample sizes of
nine locations into two collection groups. The present level and pattern of genetic diversity and structure
of Dactylorhiza hatagirea were assumed to result largely from its habitat fragmentation, its unique biological traits and evolutionary history. The genetic structure could be attributed to an earlier period of more
pronounced gene flow when the species had a more continuous distribution. A Mantel test revealed no significant positive correlation between genetic distances and geographic distances (R
2
= 0.0165, P = 0.290).
Mentha longifolia
Linn. Hudson. growing in the high altitude cold desert region of Ladakh, Jammu and Kashmir, India. The antioxidant activities of the extracts were investigated with four different methods such as 2,2’-diphenyl-1-picrylhydrazyl radical (DPPH), 2,2’-azinobis (3-ethylbenzothi-azoline-6-sulfonic acid) radical (ABTS), Reducing power assay and Ferric reducing antioxidant property (FRAP).
M . longifolia
exhibited the strongest activity as ABTS scavenger compared to DPPH and FRAP. All the assays carried out found that the plant has antioxidant potential. The total polyphenol content (TPC) and total flavonoid content of the extract was determined as 107.208 ± 34.2 mg GAE/g dry wt. and 42.47 ± 7.97 mg QE/g dry wt. respectively where 39.61% of TPC were flavonoids. A high correlation was found between the DPPH and ABTS free radical scavenging potency (R
2
= 0.869). The study has validated the medicinal potential of
Mentha longifolia
Linn. Hudson and revealed that the consumption of the plant could be beneficial as it possesses the desirable quality of antioxidant activity
Dactylorhiza hatagirea,
a critically endangered or rare terrestrial medicinal orchid in cold desert of Ladakh region. Out of the 177 bands generated from twenty random primers, 174 were polymorphic. The genetic diversity of
D. hatagirea
that was revealed by observed number of alleles (N
a
), expected number of alleles (Ne), Nei’s diversity index (H), Shannon’s diversity ind
ex (I), amplificated loci, polymorphic loci and the percentage of polymorphic loci (PPL). Pair-wise population genetic distances ranged from 0.05 to 0.23. Analysis of molecular variance (AMOVA) revealed that 57% RAPD of variability was partitioned among population.
The Principal coordinate’s analysis (PCoA) and UPGMA supported the grouping of all 96 accessions of
nine populations into four cluster groups. However, a moderate genetic differentiation among population was detected based on different measures (
Nei’s genetic diversity analysis: Gst = 0.2538;
AMOVA analysis: Fst = 0.254)
Dactylorhiza hatagirea.
Geographic variation in morphologyreflects phenotypic responses to environmental gradients and evolutionary history of populations andspecies. At points, beside its broad geographic range (Nubra, Suru and Indus valley) characterization of
Dactylorhiza
phenotype is normally accomplished by use of morphological descriptors, hence as a firststep, phenotype collection and its morphometric analysis was assessed. However, plant height, leaflength, lowermost leaf length, length of second leaf from base and mean length from lowest bract to thetop of inflorescence are presented to account for the remarkable variation in morphological characters.Tirith population showed more values of this trait while Skurru showed less value. From this, it isconcluded that Tirith showed great morphometric variation as compared to other population.Multivariate morphometric techniques, principal component analysis (PCA), multidimensional scaling(MDS) and cluster analysis were used to determine whether these populations can be reliablymorphologically similar or dissimilar. The first two principal components encompass more than 75%variation among population. The results of PCA and MDS analysis were comparable to the clusteranalysis, which shows considerable phenotypic variation in morphological and horticultural traits thatcan be utilized in its genetic improvement. To support this study, further constructive information wereprovided on the status of the populations of
D. hatagirea
which may increase the conservation value ofthis site and resolve the suitable areas with taxonomic and nomenclatural controversies.
Edgew. (Crassulaceae), well acknowledged medicinal plant is widely distributed intrans-Himalayan region of India. It has multiple uses in cuisine, forage, health care and ornamentalworth. In the present investigation, 70 wildly grown
Rhodiola
genotypes, collected from three differentsampling sites in trans-Himalayan region of Ladakh were analyzed using 40 DNA-based markers (20random amplified polymorphic DNAs (RAPDs) and 20 inter simple sequence repeats (ISSRs). RAPDanalysis yielded 134 fragments, of which 130 were polymorphic, with an average of 6.5 polymorphicfragments per primer. Of the 20 ISSR primer screened, only 8 were amplified in the presentinvestigation. These amplified primers produced 59 bands, of which 57 were polymorphic with anaverage of 7.12 polymorphic fragments per primer. RAPD markers were found more efficient withregards to polymorphism detection, as they detected 98.28% as compared to 97.55% for ISSR markers.Clustering of genotypes within groups was remained more or less same when RAPD, ISSR and RAPD +ISSR derived dendrograms were compared. The results of principal coordinates analysis (PCA) analysiswere corresponding to the dendrogram analysis. These analyses allowed us to identify the groupscorresponding to the three
Rhodiola
collection sites. Analysis of molecular variance (AMOVA) showedthat genetic variation within the populations was found maximum than among populations in all thethree cases. It may because of high level of differentiation within populations due to geographical andgenetic isolation of populations in a harsh mountainous environment. With reference to themanagement of
Rhodiola
, the high genetic separation of population indicates the requisite ofconserving the extreme possible number of populations from diverse parts of trans-Himalayas.