research article morphobiochemical variability and selection...

Research ArticleMorphobiochemical Variability and Selection Strategiesfor the Germplasm of Dactylorhiza hatagirea (D Don) SooAn Endangered Medicinal Orchid

R S Chauhan12 M C Nautiyal1 R K Vashistha1 and P Prasad1

1 High Altitude Plant Physiology Research Center HNB Garhwal University Srinagar Uttarakhand 246 174 India2 Veer Chandra Singh Garhwali (VCSG) College of Horticulture UUHF Bharsar Campus Pauri Uttarakhand 246 123 India

Correspondence should be addressed to R S Chauhan rchauhanuagmailcom

Received 6 August 2013 Revised 19 November 2013 Accepted 25 November 2013 Published 5 January 2014

Academic Editor Curtis C Daehler

Copyright copy 2014 R S Chauhan et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Dactylorhiza hatagirea (D Don) Soo (Orchidaceae) is an important endangered medicinal herb distributed in subalpine toalpine regions of the Himalayas Its tubers are important constituents of many medicines and health tonics Overexploitation formedicinal uses has decreased availability in natural habitats and this species has been enlisted as endangered making conservationand cultivation studies necessary Variability studies may serve as an important tool for effective conservation and for a cropimprovement program Therefore natural populations of D hatagirea were analyzed for variability on the basis of morphologicalbiochemical and isoenzyme patternsThe studied populations were grouped into two clusters Existing variability among differentpopulations opens up new areas for conservation and perspectives for a genetic improvement program for D hatagirea

1 Introduction

The genus Dactylorhiza (Orchidaceae) is represented byapproximately 75 species distributed in most parts of North-ern temperate zone Dactylorhiza hatagirea (D Don) Soo isa perennial orchid native to the Himalayan region [1] Thisspecies is distributed in India Pakistan Afghanistan NepalTibet and Bhutan In India it is distributed in Jammu andKashmir [2] Uttarakhand [3ndash5] and Himachal Pradesh [6]Tubers of this species are supposed as a potent medicine inLeucorrhea in Traditional System of Medicine (TSM) richsource of mucilage and are used to make Salep as foodand medicine [7] useful as expectorant and astringent [8]Recently herbal health beverages have also been developedfrom this species [9] The species has been categorized asendangered [10] due to overexploitation of tubers formedici-nal value habitat degradations and other biotic interferencesin its distribution ranges The market demand of this speciesis increasing while supply is gradually decreasing [11] that callfor conservation as well as cultivation

Effective conservationmanagement and recovery of rareand endangered species can be achieved through variability

analysis Variations in morphological appearance among themembers of a species become important to the biologistsolely because they indicate the presence or absence ofphysiological attributes that are of ecological significanceStatisticalmethods including principle components or clusteranalysis can be used as useful tools for screening of suchpopulations [12] Adequate information is available on thedistribution pattern [4 5] medicinal properties [7ndash9] andpropagation methods [13] of D hatagirea but there is alack of knowledge about morphological biochemical andgenetic variability In view of above present paper is focusedon detail investigation on morpho-biochemical variabilityamong different populations ofD hatagirea Such studies willbe helpful for effective conservationmanagement and geneticimprovement of D hatagirea

2 Materials and Methods

Field survey was conducted in several parts of UttarakhandIndia known for occurrence of D hatageria and ninedifferent populations were selected for detail investigations

Hindawi Publishing CorporationJournal of BotanyVolume 2014 Article ID 869167 5 pageshttpdxdoiorg1011552014869167

2 Journal of Botany

31∘30

998400

31∘00

998400

30∘00

998400

30∘30

998400

29∘30

998400

29

10 0 10 20

∘00

998400

31∘30

998400

31∘00

998400

30∘00

998400

30∘30

998400

29∘30

998400

29∘00

998400

78∘00

99840078

∘30

99840079

∘00

99840079

∘30

99840080

∘00

99840080

∘30

99840081

∘00

998400

78∘00

99840078

∘30

99840079

∘00

99840079

∘30

99840080

∘00

99840080

∘30

99840081

∘00

998400

(km)

1

23

4

5

67

8 9

Figure 1 Map showing locations of selected sites ofD hatagirea (1 DH1 2 DH2 3- DH3 4 DH4 5 DH5 6 DH6 7 DH7 8 DH8 9 DH9)

Table 1 General characteristic of selected natural habitats of D hatagirea

Study sites Code Distribution range (msl) Habitat Latitude LongitudeDayara (Uttarkashi) DH1 3000ndash3400 Open meadow 30∘501015840N 78∘331015840EDronagiri (Chamoli) DH2 3200ndash3500 Open meadow 30∘301015840N 79∘521015840EKedarnath (Rudraprayag) DH3 2900ndash3500 Open meadow 30∘441015840N 79∘031015840EKunwari Pass (Chamoli) DH4 3000ndash3400 Open meadow 30∘071015840N 79∘581015840EMadhyameshwar (Rudraprayag) DH5 3500ndash3800 Open meadow 30∘391015840N 79∘141015840EPanwalikantha (Tehri) DH6 3300ndash3500 Open meadow 30∘341015840N 78∘521015840ERudranath (Chamoli) DH7 3000ndash3200 Open meadow 30∘111015840N 79∘281015840ETungnath (Rudraprayag) DH8 3200ndash3600 Open meadow 30∘141015840N 79∘221015840EThe Valley of Flowers (Chamoli) DH9 3000ndash4200 Open meadow 30∘101015840N 79∘571015840E

A detailed description of selected study sites is presented(Figure 1 Table 1) Considering endangered status of thespecies in Uttarakhand [10] experiments were designed insuch a way that minimum number of samples may provideoptimum information Ten mature plants from each studysite were sampled randomly for morphological details (plantheight number of flowerplant inflorescence length andeconomic yield) during themonth of SeptemberThese plantswere dug out for the estimation of economic yield (tubers)The tubers were separated washed with running water anddried at 40∘C temperature until constant weight One set offresh tubers was immediately crushed in liquid nitrogen toseize enzymatic activities and then stored at (minus80∘C) in a deepfreezer until analysis

These samples were used to estimate variations in solu-ble sugars protein content and isoenzymes Soluble sugarcontent was estimated using the Anthrone method [14] andsoluble protein content was determined using the Bradfordmethod [15] The absorbance of the reaction mixture wasread in a spectrophotometer (Beckman DU-640) Isoenzymevariation was analyzed on 10 polyacrylamide slab gels in

a discontinuous gel electrophoretic system at a constantcurrent of 20mA [16] Esterase (EC 3111) and peroxidase(EC 3411) isoenzymes were detected in gels using themethods of Bhadula and Sawhney [17] and Welter [18]respectively among different populations Collected datawere subjected to analysis of variance (ANOVA) and onlysignificant variations were used for multivariate analysis ofMahalanobis distance [19]The analysis was performed usingindostat statistical package (httpwwwwindostatorg) andthe cluster formation was confirmed by the Tocher method[20]

3 Results

The germplasm collected from different populations showedconsiderable variation in plant height number of flowerslength of inflorescence economic yield and biochemicalparameters (Table 2) The variability range was between 132and 543 cm in plant height 149 and 294 in number offlowers per plant 39 and 117 cm in inflorescence length and

Journal of Botany 3

Table 2 Mean growth performance of D hatagirea at different natural populations

Study sites Economic yield(gplant)

Plant height(cm)

Number offlowersplants

Length ofInflorescence (cm)

Soluble sugar(mgg)

Starch(mgg)

Solubleprotein (mgg)

DH1 05 450 220 108 206 456 106DH2 06 355 223 100 340 548 268DH3 07 138 149 39 345 571 115DH4 05 132 245 65 356 560 104DH5 06 372 265 95 280 591 143DH6 05 543 266 96 335 570 118DH7 05 513 294 117 407 681 104DH8 04 410 234 107 217 532 156DH9 04 290 220 57 230 743 269Mean 05 356 235 86 302 584 154SDlowast 01 146 41 26 71 84 68Range 04ndash07 132ndash543 149ndash294 39ndash117 206ndash407 456ndash743 104ndash269CD 5lowastlowast 018 999 572 473 958 855 487lowastSD standard deviation lowastlowastCD critical difference

50 100 150 200

1

8

5

6

2

9

3

4

7

Cluster 1

Cluster 2

Figure 2 Dendrogram of D hatagira by Tocher method (1 DH1 2DH2 3 DH3 4 DH4 5 DH5 6 DH6 7 DH7 and 8 DH8 9 DH9)

04 and 07 g per plant in economic yield Biochemical param-eters also showed great variability among studied popula-tions The variability range was between 206 and 407mggin soluble sugar 456 and 743mgg in starch and 104 and269mgg in soluble protein content Such variations couldbe due to edaphic microclimate environmental factors andso forth In view of considerable amount of variability 1198632values were compared for all possible pairs of populationsBased on 1198632 values these populations could be grouped intotwo clusters (Figure 2)The higher contribution of charactersregarding variability was shown by soluble protein (7222)plant height (1389) and starch content (833) whereasother quantitative characters that is number of flowersand soluble sugar showed similar share of 278 Meangrowth performance of both clusters is presented separatelyin Table 3 Cluster one contains maximum 6 populationsthat is DH1 DH8 DH5 DH6 DH2 and DH9 whereascluster two contains 3 population DH3 DH4 and DH7 Theaverage intracluster distance ranged from 7 to 8 whereas theintercluster distance ranged from 8 to 13 (Table 4)

There were significant differences among the parame-ters measured indicating variability among the populations

(Figure 3(a)) Total number intensity and position of bandsvaried from population to population DH8 and DH4 pop-ulations emerged as a distinct cluster by showing specificbanding pattern of esterase isoenzyme as compared to therest of the populations These two populations showed morecompact and dark stained band The peroxidase pattern inDH3 and DH7 populations emerged as a distinct clusture byshowing specific banding pattern Intensity of bands was verydark in DH3 followed by DH7 populations whereas bandsof light intensity were resolved in DH1 and DH2 population(Figure 3(b))

4 Discussion

Germplasm of D hatageria collected from different geo-graphical locations of the Garhwal Himalaya was groupedinto two clusters based on their similarities and variationsThey varied considerably in the morphology biochemicalparameters and isoenzyme pattern Over the long term theability of a population to respond adaptively to environmentalchanges depends on the level of genetic variability or diversityit contains [21] Variations in plant height were related to thesoil and climatic factors [22] Variation in the morphologicalcharacters is possible due to variation in topography ele-vation soil rainfall and other climatic conditions peculiarto Himalaya [23] In addition available soil nutrient levelalso plays an important role in determining morphologicalvariations in plants [24 25] and climatic factors can beconsidered as indicators to alter assimilate investment pattern[26]

Biochemical variations suggested polymorphic natureof the species and it is argued that they can be used asan indicators to the impact of microclimatic conditionson plant life [25] and therefore reflects an adaptive fea-ture of the species [27] Morphological variability in plantsand differences in protein and sugar levels were importantparameters to study natural populations of rare plants these

4 Journal of Botany

Table 3 Cluster mean of seven characters of D hatagirea

Clusters Number oflocations

Economic yield(gplant)

Plant height(cm)

Number offlowersplant

Inflorescencelength (cm)

Soluble sugar(mgg)

Starch(mgg)


1 6 05 4032 2381 927 268 5731 17662 3 055 2609 2291 737 3694 604 1078

Table 4 Intra- and intercluster divergence (1198632) among threeclusters of D hatagirea

Clusters 1 21 752 1340

851

DH1 DH4 DH6 DH5 DH9 DH10 DH8 DH7 DH3 DH2

(a)


(b)

Figure 3Heterozygosity in isoenzyme banding pattern (a) Esteraseand (b) peroxidase among ten different populations of D hatagirea

parameters also reflect a variation due to environmentalconditions Average economic yield in DH2 population ispositively correlated with the number of flowers solublesugar starch and soluble protein content Such informationon morphometric primary metabolites and isoenzyme pat-tern suggests that secondary metabolites may also vary inthese populations Increase in primary metabolites showedsignificant increase in active constituents in adantoid ferns[28] Isoenzyme studies also supported variability amongstudied populations Germplasm diversity based on standardmorphological marker has proved to be inadequate because

of wide spectrum of phenotypic variation and their inter-action with environment [29] whereas esterase isoenzymeis an excellent marker in variability studies which indicatesintrapopulation variations [30]

5 Conclusions

This study revealed significant differences among the param-eters measured indicating variability among these popula-tions Variability existing among different populations opensa new area for conservation and genetic improvement of Dhatagirea These variations may be used as markers for cropimprovement program DH2 (Dronagiri) population showedbetter performance among the studied populations whichcan be used as elite germplasm source for multiplication ofDhatagirea On the basis of such multifaceted information onspecific species future conservation strategies and cultivationof wild medicinal species can be initiated [31]

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors are thankful to Dr H Purohit (SRF) HAPPRCSrinagar for helping in this work Financial support fromDepartment of Biotechnology Government of India NewDelhi is gratefully acknowledged

References

[1] H K Badola and S Aitken ldquoTheHimalayas of India a treasuryof medicinal plants under siegerdquo Biodiversity vol 4 pp 3ndash132003

[2] U Dhar and P Kachroo Alpine Flora of Kashmir HimalayaScientific Publishers Jodhpur India 1983

[3] P K Hajra and B Badoni PlantWealth of NandaDevi BiosphereReserve Botanical Survey of India Calcutta India 1995

[4] A Bhatt S K Joshi and S Gairola ldquoDactylorhiza hatagirea (DDon) Soomdasha west Himalayan orchid in perilrdquo Current Sciencevol 89 no 4 pp 610ndash612 2005

[5] R S Chauhan and M C Nautiyal ldquoA note on Dactylorhizahatagirea (D Don) Soordquo The McAllen International OrchidSociety Journal vol 9 no 2 pp 8ndash12 2008

[6] B S Aswal and B N Mehrotra Flora of Lahaul-Spiti BishenSingh and Mahendra Pal Singh Dehradun India 1994

[7] CSIRTheWealth of India vol 10 of Publication and InformationDirectorate Council of Scientific and Industrial Research NewDelhi India 1976

Journal of Botany 5

[8] K R Kirtikar and B D Basu Indian Medicinal Plants BishenSingh Mahendra Pal Singh Dehradun India 1989

[9] B Ballabh O P Chaurasia and Z Ahmed ldquoHerbal productsfrom high altitude plants of LadakhHimalayardquoCurrent Sciencevol 92 no 12 pp 1664ndash1665 2007

[10] AnonConservation Assessment andManagement Prioritizationfor the Medicinal Plants of Jammu and Kashmir HimachalPradesh and Uttarakhand FRLHT Bangalore India 2003

[11] D K Ved and G S Goraya Demand and Supply of MedicinalPlants in India Bishan SinghMahendra Pal Singh Dehradun ampFRLTH Bangalore India 2008

[12] H R Karimi Z Zamani A Ebadi andM R Fatahi ldquoMorpho-logical diversity of Pistacia species in Iranrdquo Genetic Resourcesand Crop Evolution vol 56 no 4 pp 561ndash571 2009

[13] D Giri and S Tamta ldquoCombined effect of PGRs and soil facili-tate early flowering of an endangered alpine orchidDactylorhizahatagirea at lower elevationrdquo Current Science vol 99 no 1 pp21ndash23 2010

[14] R M McCready J Guggolz V Silviera and H S OwensldquoDetermination of starch and amylose in vegetables applicationto peasrdquoAnalytical Chemistry vol 22 no 9 pp 1156ndash1158 1950

[15] M M Bradford ldquoA rapid and sensitive method for the quanti-tation of microgram quantities of protein utilizing the principleof protein dye bindingrdquoAnalytical Biochemistry vol 72 no 1-2pp 248ndash254 1976

[16] B J Davis ldquoDisc electrophoresis II Method and applicationto human serum proteinsrdquo Annals of the New York Academy ofSciences vol 121 pp 404ndash427 1964

[17] S K Bhadula and V K Sawhney ldquoEsterase activity andisozymes during the ontogeny of stamens of male fertile Lycop-ersicon esculentummill a male sterile stamenless-2 mutant andthe low temperature-revertedmutantrdquo Plant Science vol 52 no3 pp 187ndash194 1987

[18] L R Welter ldquoIsoenzyme analysis of cultured plant cellsrdquo inPlant Tissue Culture Methods L R Welter and F ConstabelEds pp 105ndash111 National Research Council of Canada Saska-toon Canada 1982

[19] P C Mahalanobis ldquoOn the generalized distance in statisticsrdquoProceedings of the National Institute of Science vol 2 pp 49ndash551936

[20] C R Rao Advanced Statistical Methods in Biomedical ResearchJohn Wiley amp Sons New York NY USA 1952

[21] F J Ayala and J A Kiger Modern Genetics BenjaminCum-mings Menlo Park Calif USA 2nd edition 1984

[22] N Krishnan A Jeyachandran and N Nagendran ldquoEffect ofseasonal and altitudinal variations on growth performance ofAcalypha indica Linn in Alagar Hill (Eastern Ghats) SouthIndiardquo Tropical Ecology vol 41 no 1 pp 41ndash45 2000

[23] R D Gaur D S Rawat and L R Dangwal ldquoA contributionto the flora of Kunwari Pass-Dalisera alpine zone in GarhwalHimalayardquo Journal of Economic and Taxonomic Botany vol 19pp 9ndash26 1995

[24] M Pigliucci P Diiorio and C D Schlichting ldquoPhenotypicplasticity of growth trajectories in two species of Lobelia inresponse to nutrient availabilityrdquo Journal of Ecology vol 85 no3 pp 265ndash276 1997

[25] C P Kuniyal S K Bhadula and P Prasad ldquoMorphologicaland biochemical variations among the natural populationsof Aconitum atrox (Bruhl) Muk (Ranunculaceae)rdquo Journal ofPlant Biology vol 29 no 1 pp 91ndash96 2002

[26] C Korner and S P Menendez-Riedl ldquoThe significance ofdevelopmental aspects in plant growth analysisrdquo in Causes andConsequences of Variation in Growth Rate and Productivityof Higher Plants H Lambers Ed pp 141ndash157 AcademicPublishers The Hague The Netherlands 1989

[27] S K Bhadula A P Thapliyal and A N Purohit ldquoSeed proteinesterase and acid phosphatase in Rhododendron species fromdifferent altitudes in Garhwal Himalayardquo Indian Journal ofExperimental Biology vol 19 pp 199ndash200 1981

[28] P Guha K Gupta and R Mukhopadhyay ldquoImpact of seasonson some biochemical parameters in three adantoid fernsrdquoIndian Journal of Plant Physiology vol 11 pp 152ndash159 2006

[29] L Mannetji ldquoConsiderations on the taxonomy of the genusStylosanthesrdquo in The Biology and Agronomy of StylosanthesH M Stace and L A Edye Eds Academic Press SydneyAustralia 1984

[30] S K Bhadula A Singh H Lata C P Kuniyal and A NPurohit ldquoGenetic resources of Podophyllum hexandrum Roylean endangered medicinal species from Garhwal HimalayaIndiardquo Plant Genetic Resources Newsletter vol 106 pp 26ndash291996

[31] S Airi R S Rawal U Dhar and A N Purohit ldquoAssessmentof availability and habitat preference of Jatamansimdasha criticallyendangeredmedicinal plant of west HimalayardquoCurrent Sciencevol 79 no 10 pp 1467ndash1471 2000

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology

2 Journal of Botany

31∘30

998400

31∘00

998400

30∘00

998400

30∘30

998400

29∘30

998400

29

10 0 10 20

∘00

998400

31∘30

998400

31∘00

998400

30∘00

998400

30∘30

998400

29∘30

998400

29∘00

998400

78∘00

99840078

∘30

99840079

∘00

99840079

∘30

99840080

∘00

99840080

∘30

99840081

∘00

998400

78∘00

99840078

∘30

99840079

∘00

99840079

∘30

99840080

∘00

99840080

∘30

99840081

∘00

998400

(km)

1

23

4

5

67

8 9

Figure 1 Map showing locations of selected sites ofD hatagirea (1 DH1 2 DH2 3- DH3 4 DH4 5 DH5 6 DH6 7 DH7 8 DH8 9 DH9)

Table 1 General characteristic of selected natural habitats of D hatagirea

Study sites Code Distribution range (msl) Habitat Latitude LongitudeDayara (Uttarkashi) DH1 3000ndash3400 Open meadow 30∘501015840N 78∘331015840EDronagiri (Chamoli) DH2 3200ndash3500 Open meadow 30∘301015840N 79∘521015840EKedarnath (Rudraprayag) DH3 2900ndash3500 Open meadow 30∘441015840N 79∘031015840EKunwari Pass (Chamoli) DH4 3000ndash3400 Open meadow 30∘071015840N 79∘581015840EMadhyameshwar (Rudraprayag) DH5 3500ndash3800 Open meadow 30∘391015840N 79∘141015840EPanwalikantha (Tehri) DH6 3300ndash3500 Open meadow 30∘341015840N 78∘521015840ERudranath (Chamoli) DH7 3000ndash3200 Open meadow 30∘111015840N 79∘281015840ETungnath (Rudraprayag) DH8 3200ndash3600 Open meadow 30∘141015840N 79∘221015840EThe Valley of Flowers (Chamoli) DH9 3000ndash4200 Open meadow 30∘101015840N 79∘571015840E

A detailed description of selected study sites is presented(Figure 1 Table 1) Considering endangered status of thespecies in Uttarakhand [10] experiments were designed insuch a way that minimum number of samples may provideoptimum information Ten mature plants from each studysite were sampled randomly for morphological details (plantheight number of flowerplant inflorescence length andeconomic yield) during themonth of SeptemberThese plantswere dug out for the estimation of economic yield (tubers)The tubers were separated washed with running water anddried at 40∘C temperature until constant weight One set offresh tubers was immediately crushed in liquid nitrogen toseize enzymatic activities and then stored at (minus80∘C) in a deepfreezer until analysis

These samples were used to estimate variations in solu-ble sugars protein content and isoenzymes Soluble sugarcontent was estimated using the Anthrone method [14] andsoluble protein content was determined using the Bradfordmethod [15] The absorbance of the reaction mixture wasread in a spectrophotometer (Beckman DU-640) Isoenzymevariation was analyzed on 10 polyacrylamide slab gels in

a discontinuous gel electrophoretic system at a constantcurrent of 20mA [16] Esterase (EC 3111) and peroxidase(EC 3411) isoenzymes were detected in gels using themethods of Bhadula and Sawhney [17] and Welter [18]respectively among different populations Collected datawere subjected to analysis of variance (ANOVA) and onlysignificant variations were used for multivariate analysis ofMahalanobis distance [19]The analysis was performed usingindostat statistical package (httpwwwwindostatorg) andthe cluster formation was confirmed by the Tocher method[20]

3 Results

The germplasm collected from different populations showedconsiderable variation in plant height number of flowerslength of inflorescence economic yield and biochemicalparameters (Table 2) The variability range was between 132and 543 cm in plant height 149 and 294 in number offlowers per plant 39 and 117 cm in inflorescence length and

Journal of Botany 3



Plant height(cm)



Soluble sugar(mgg)

Starch(mgg)



50 100 150 200

1

8

5

6

2

9

3

4

7

Cluster 1

Cluster 2





4 Discussion



4 Journal of Botany




Plant height(cm)



Soluble sugar(mgg)

Starch(mgg)


1 6 05 4032 2381 927 268 5731 17662 3 055 2609 2291 737 3694 604 1078


Clusters 1 21 752 1340

851


(a)


(b)




5 Conclusions




Acknowledgments


References








Journal of Botany 5
































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Journal of Botany 3



Plant height(cm)



Soluble sugar(mgg)

Starch(mgg)



50 100 150 200

1

8

5

6

2

9

3

4

7

Cluster 1

Cluster 2





4 Discussion



4 Journal of Botany




Plant height(cm)



Soluble sugar(mgg)

Starch(mgg)


1 6 05 4032 2381 927 268 5731 17662 3 055 2609 2291 737 3694 604 1078


Clusters 1 21 752 1340

851


(a)


(b)




5 Conclusions




Acknowledgments


References








Journal of Botany 5
































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

4 Journal of Botany




Plant height(cm)



Soluble sugar(mgg)

Starch(mgg)


1 6 05 4032 2381 927 268 5731 17662 3 055 2609 2291 737 3694 604 1078


Clusters 1 21 752 1340

851


(a)


(b)




5 Conclusions




Acknowledgments


References








Journal of Botany 5
































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Journal of Botany 5
































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

research article morphobiochemical variability and selection...

Documents