Industrial Crops and Products 50 (2013) 112–117

Contents lists available at SciVerse ScienceDirect

Industrial Crops and Products

journal homepage: www.elsevier.com/locate/indcrop

Altitudinal variability in anthraquinone constituents from novel

cytotypes of Rumex nepalensis Spreng—a high value medicinal herb of
North Western Himalayas
Umer Farooq a,1 , Shahzad A. Pandith b,1 , Manjit Inder Singh Saggoo a,∗∗ ,
Surrinder K. Lattoo b,∗
a
Department of Botany, Punjabi university, Patiala, Punjab 147002, India
b
Plant Biotechnology, Indian Institute of Integrative Medicine (CSIR), Canal Road, Jammu Tawi 180001, India

a r t i c l e i n f o a b s t r a c t

Article history: Rumex nepalensis is a reputed medicinal herb of North Western Himalayas. It owes its pharmacological
Received 19 February 2013 significance to an active class of compounds known as anthraquinones. In the present study four differ-
Received in revised form 21 June 2013 ent intraspecific cytotypes with varying ploidy level ranging from tetraploidy (4×, 2n = 40) to octaploidy
Accepted 28 June 2013
(8×, 2n = 80) were discovered, hitherto unreported. The investigation also included the evaluation of
chemical variability in four major anthraquinone constituents of different cytotypes from five altitudi-
Keywords:
nal habitats of Himalayas using standard HPLC method. There was significant increase in the aglycone
Rumex nepalensis
components of anthraquinones with the increase in ploidy status, except for dodecaploid type (12×,
Anthraquinones
Cytotype
2n = 120) that showed a decreasing trend in their accumulation. However, the overall concentration
Meiotic analysis of anthraquinones, both glycone and aglycone constituents was found to be highest (33.95%) in the
HPLC dodecaploid type from Lidderwatt location. Polyploidy, an important driver of eukaryotic evolution is a
Polyploidy prevalent feature among angiosperm species providing dynamic genome flexibility to them. Polyploids
can undergo rapid structural and functional alterations, allowing them to adapt and persist across het-
erogeneous landscapes in the long run. It is reflected in the robust adaptability of different cytotypes of
R. nepalensis. These display appreciable chemical variability, and ecological plasticity possibly because
of the intraspecific polyploidization in response to different ecological niches of a geographical region.
The scope of present investigation also extends to identify elite cytotypes in terms of their desirable
chemoprofiles for pharmaceutical and commercial purposes.
© 2013 Elsevier B.V. All rights reserved.

1. Introduction
Rumex nepalensis Spreng (Polygonaceae) is a perennial, ascend-
ing herb distributed throughout Himalayas from Bhutan to Kashmir
and also to Turkey, Java and South Africa. It is a fairly common
plant of higher altitudes and grows between 900–4000 m on moist
as well as dry slopes, under shades, and even in plains throughout
Kashmir (India). R. nepalensis is highly reputed for various thera-
peutic purposes in Indian traditional systems of medicine. The root
of R. nepalensis is purgative and is used as a substitute for Rheum
species (Manandhar, 2002). A strong decoction is applied to dislo-
cated bones. Root paste is applied externally to relieve headache.
∗ Corresponding author. Tel.: +91 9419203465; fax: +91 191 2569019.
∗∗ Corresponding author.
E-mail addresses: msaggoo@gmail.com (M.I. Singh Saggoo), sklattoo@iiim.ac.in
(S.K. Lattoo).
1
Authors have contributed equally.
Its leaves are antiseptic and are used for the treatment of syphilic
and colic ulcers (Kirtikar and Basu, 1987). Several anthraquinones,
naphthalenes, flavonoids and other phenolic compounds have been
reported from R. nepalensis (Liang et al., 2010). These compounds
are used for the treatment of bleeding, tumour, inflammation,
pain, constipation and tinea in the Chinese folk medicine (Zhang
et al., 2008). R. nepalensis has been investigated for antihistaminic,
anticholinergic, antibradykinin, antiprostaglandin (Aggarwal et al.,
1986), purgative, antibacterial (Ghosh et al., 2003), antipyretic
(Venkatesh et al., 2003) and anti-inflammatory activities (Gautam
et al., 2010). The antibacterial, antifungal and insecticidal proper-
ties have also been reported from the extracts of this plant (Hussain
et al., 2010).
In present investigation our endeavour was to explore chemi-
cal diversity in R. nepalensis. It also included cytological analysis of
various populations from Kashmir Himalayas to determine chro-
mosome counts in corroboration with comparative chemoprofiles
of the five different cytotypes of R. nepalensis employing standard
HPLC method (Singh et al., 2005) with slight modifications. The

0926-6690/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.indcrop.2013.06.044
 U. Farooq et al. / Industrial Crops and Products 50 (2013) 112–117
Table 1
Chromosome count and ploidy status recorded in different populations of Rumex nepalensis from five locations of Kashmir Himalayas.
S. No. Meiotic chromosome no. (‘n’) Cytotype (ploidy status)
b ◦
1 20 Tetraploid (4x = 40)
2 30b Hexaploid (6x = 60)
3 40b Octaploid (Y) (8x = 80)
4 40b Octaploid (T) (8x = 80)
5 60 Dodecaploid (12x = 120)
a
Herbarium code of the Botany Department, Punjabi University, Patiala, as per “Index Herbariorum” by Holmgren and Holmgren (1998).
b
First chromosomal report at world level.
investigation suggests positive correlation between ploidy level
and metabolite accumulation in relation to different altitudinal gra-
dients. This throws a fresh perspective to identify elite cytotypes
in terms of their desirable chemoprofiles for pharmaceutical and
commercial purposes.
2. Materials and methods
2.1. Plant materials and chemicals
The wild growing plants of R. nepalensis were collected from five
different locations of the Kashmir valley (J&K state, India). The plant
collections were identified by Dr. S. K. Srivastava, in-charge Herbar-
ium Botanical Survey of India (BSI), Northern circle, Dehradun
(India). The voucher specimens of the plants of different popu-
lations were deposited in the Herbarium, Department of Botany,
Punjabi University, Patiala, India (Table 1). Pure standards of two
major anthraquinones namely emodin and chrysophanol were pur-
chased from Sigma–Aldrich (St. Louis, USA) and their respective
glycosides, emodin glycoside and chrysophanol glycoside (Fig. 1)
were obtained from Institute of Himalayan Bioresource Technol-
ogy, Palampur India (kindly provided by A. K. Sinha). Their purities
were above 99%, as determined by HPLC analysis. All chemicals and
solvents used were have analytical and HPLC grade (E. Merck, Mum-
bai, India). Fresh ultra-pure distilled water with resistivity greater
than 18 M was used.
2.2. Meiotic analysis
For meiotic analysis the young floral buds at an ideal stage were
fixed in freshly prepared carnoy’s fixative (6 alcohol: 3 chloroform:
1 acetic acid v/v/v) for 24 h and then preserved in 70% alcohol at
4 ◦ C in the refrigerator. The cytological preparations were made
using the squash technique in 2% acetocarmine (Rana et al., 2012).
A total of 45 pollen mother cells (PMC) from temporary slides were
examined to determine the exact chromosome number at different
stages of meiosis. Photomicrographs of chromosome counts were
taken from freshly prepared slides using Nikon 80i Eclipse micro-
scope. For previous chromosome reports, in addition to the IPCN
database (http://www. tropicos.org) various indices to plant chro-
mosome numbers were consulted (Darlington and Wylie, 1995;
Fig. 1. Chemical structures of major anthraquinones and their glycosides.
113
Location of plant collection Accession number (PUN)a
 ◦ 
Zawoora (33 43 N, 75 14 E; 2000 m) 57,691
Zawoora upper reaches (34◦ 44 N, 74◦ 48 E; 2300 m) 57,676
Yusmarg (33◦ 47 N, 74◦ 39 E; 2600 m) 57,677
Thajwas (34◦ 18 N, 74◦ 48 E; 3400 m) 57,660
Lidderwatt (34◦ 04 N, 75◦ 17 E; 3500 m) 57,683
Moore, 1967–74; Kumar and Subramanian, 1986; Khatoon and Ali,
1993).
2.3. Chemical evaluation
Leaf samples of five cytotypes of R. nepalensis collected from
five different locations of the Kashmir valley were air-dried and
ground into fine powder with a mortar and pestle. The powdered
leaves (50 g of each sample) of five different plant samples were
extracted each with methanol (3 × 300 mL) using Soxchlet extrac-
tion. The extracts were filtered and solvents were removed under
reduced pressure. For arbitrary evaluation of the extracted sam-
ples, TLC with solvent compositions of 5%, 10% and 15% methanol
in dichloromethane were taken. This showed marked variation in
the Rf (retention factor) values and intensity of the visible spots,
indicating presence of various compounds in different concentra-
tions (Fig. 2). Moreover, TLC spots were non-UV active. The stock
solutions (1 mg/mL) of each of the four anthraquinones were freshly
prepared in a mixture of methanol: acetonitrile (80:20, v/v). Sim-
ilarly, the crude extracts of five cytotypes were also dissolved in
the same solvent composition (20 mg/mL) and were filter steril-
ized with 0.45 ␮m membrane filters (Millipore, Bedford, USA). The
HPLC (Shimadzu CLASS-VP V 6.14 SPI model) equipped with RP-
18e column (E-Merck, 5 ␮m, 4.6 × 250 nm), a photo-diode array
detector (SPD-M10A VP model) and a pump (LC-10AT VP model)
was used for analysis of hydroxy-anthraquinone derivatives. Ace-
tonitrile: methanol (95:5, v/v) (solvent B) and water: acetic acid
(99.9:0.1, v/v, pH 3.5) (solvent A) were used as mobile phase with
a linear gradient elution as follows: 0–15 min, 20% B; 15–25 min,
20% B; 25–30 min, 50% B; 30–40 min, 70% B; 40–45 min, 100% B;
45–50 min, 20% B; 50–55 min, 20% B; at a flow rate of 0.8 ml/min.
The detection wavelength was set at 254 nm. Injection volume of
the sample was 10 ␮l and the column temperature 30 ◦ C (Fig. 3).
3. Results and discussion
Detailed cytological analysis of various populations of R.
nepalensis revealed the existence of four different chromosomal
races with meiotic chromosome numbers n = 20, 30, 40 and 60 in
the Kashmir region (Table 1). These chromosome counts are the
first ever records for this species. The present chromosome count
of n = 60 (2n = 12x= 120) confirms the previous record (Degraeve,
114 U. Farooq et al. / Industrial Crops and Products 50 (2013) 112–117

Fig. 2. TLC profiles of methanolic extracts of different cytotypes of R. nepalensis in 5% (a), 10% (b) and 15% (c) methanol in dichloromethane showing various compounds,
found to be UV inactive. Abbreviations: 1 = Tetraploid; 2 = Hexaploid; 3 = Dodecaploid; 4 = Octaploid (Y) and 5 = Octaploid (T).

Fig. 3. HPLC chromatograms of standard major anthraquinones (a) and methanolic extracts of leaves of tetraploid (b), hexaploid (c), octaploid Y (d), octaploid T (e) and
dodecaploid (f) cytotypes of R. nepalensis. Abbreviations: 1 = emodin; 2 = chrysophanol; 3 = emodin glycoside; 4 = chrysophanol glycoside.
 U. Farooq et al. / Industrial Crops and Products 50 (2013) 112–117 115

Fig. 4. Meiotic chromosome numbers in five populations from different altitudinal ranges in R. nepalensis; A PMC at metaphase- I showing twenty bivalents (n = 20; Zawoora,
2000 m in altitude) (a); A PMC showing thirty bivalents (n = 30; Zawoora, 2300 m) (b); A PMC at metaphase- I showing forty bivalents (n = 40, Yusmarg, 2600 m) (c); A PMC
at metaphase-I showing 40 bivalents (n = 40, Thajwas, 3400 m) (d); A PMC at diakinesis showing sixty bivalents (n = 60, Lidderwatt (3500 m) (e). Bar = 10 ␮m.

1975). The basic chromosome numbers for the genus Rumex are A well marked increasing trend in the concentration of all four
reported as 7, 8, 9, and 10 (Darlington and Wylie, 1995). Conform- compounds from tetraploid to octaploid variants is in conformity
ing to x = 10, the present investigated taxa are tetraploid (Fig. 4a), with previous reports wherein it has been shown that increase in
hexaploid (Fig. 4b), octaploid (Fig. 4c and d) and dodecaploid secondary metabolite content is positively correlated with ploidy
(Fig. 4e) respectively. Different ploidy levels and wide range of level and altitude (Zidorn, 2010; Lavania et al., 2012). Compara-
ecological plasticity explains the wide distribution of this group tive chemoprofile of two octaploids collected from two different
in North Western Himalayas. The occurrence of intraspecific poly- locations designated as octaploid (T) and octaploid (Y) respectively
ploids in R. nepalensis is indicative of the fact that its genome (Table 1), showed an appreciable difference in the concentration
is still in flux and constant evolution, possibly to enhance the of anthraquinones (Fig. 5). This can be ascribed to the effect of
adaptive and survival value of the species under varied ecological altitudinal gradients on the secondary metabolite production. Sim-
niches of Himalayan region (Lattoo et al., 2006a). The new cyto- ilar pattern of metabolite accumulation has also been reported in
types/polyploids in all probability have high chance of survival at Lupinus argenteus (Carey and Wink, 1994) and Ginkgo biloba (Kaur
different altitudinal gradients. et al., 2012). It is well documented in literature that metabolite
Plants produce an enormous diversity of low molecular weight accumulation is a function of interaction between genetic consti-
compounds with a wide range of activity. However this number tution and environmental variables (Lattoo et al., 2006b; Kooke
gets further increased by multiple decorations of the common
skeleton by acylation, methylation, hydroxylation or conjugation
reactions used for various metabolic activities in the plant. One
of these most widespread modifications is glycosylation which
is related to specific plant functions like xenobiotic detoxifica-
tion, regulation of hormone homeostasis, and biosynthesis and
storage of secondary compounds (Gachon et al., 2005). The over-
all concentration of glycosides was found to be higher than that
of their respective aglycone forms at all ploidy levels. The dode-
caploid type from Lidderwatt (34◦ 04 N, 75◦ 17 E; 3500 m in altitude)
showed the maximum accumulation of emodin and chrysophanol
glycosides than their corresponding aglycone moieties. This shuf-
fle in the direction of glycosylation, like many of the plant natural
products may be towards enhancing their solubility and stabil-
ity for facilitating their easy storage and accumulation in plant
cells. Among the four major anthraquinones, emodin glycoside
(38.071 mg/g ± 0.907) was found to be in highest concentration
Fig. 5. Graphic representation of the concentrations of key anthraquinones in five
followed by emodin (5.631 mg/g ± 0.235), chrysophanol glyco-
different cytotypes of R. nepalensis. All values obtained were means of triplicate with
side (4.524 mg/g ± 0.268) and chrysophanol (4.262 mg/g ± 0.175). standard errors.
116 U. Farooq et al. / Industrial Crops and Products 50 (2013) 112–117

Table 2
Relative percentage of four key anthraquinone constituents in the leaves of five different cytotypes of Rumex nepalensis.

Ploidy Level Tetraploid Hexaploid Octaploid (Y) Octaploid (T) Dodecaploid

Relative percentage (%)a 2.73 8.49 28.98 25.85 33.95

a
Concentration of all four anthraquinones in one cytotype/total concentration in all cytotypes × 100.

and Keurentjes, 2012). It is an efficient adaptive strategy employed
by the plants to cope up with the effect of many biotic and abi-
otic stresses to perpetuate in diverse environments. The variability
of major phyto-constituents within the same species at different
altitudes and temperature ranges present a significant relation-
ship between the quality and quantity of active principles. Changes
in the chemical constituents of Achillea millefolium has also been
recorded from different Himalayan habitats due to the interaction
of plant populations to diverse habitats within a geographic area as
primary selection factors give rise to major or minor differences
in the content of chemical constituents (Agnihotri et al., 2005).
In many cases, it may also influence the therapeutic potential of
medicinal plants (Zarinkamar et al., 2011).
The relative percentage of different chemical constituents is a
useful criterion to evaluate the total content of compounds present
in a given entity. In terms of relative percentage, anthraquinones
were found to be maximum (33.95%) in the dodecaploid type and
minimum (2.73%) in the tetraploid one, providing a clue towards
the difference in pharmacological significance of these two cyto-
types (Table 2).
Morphological variations in all the populations of five different
cytotypes were evaluated. The tetraploid plants which measured
120–150 cm in size were tallest than the rest of cytotypes. There
was profuse secondary branching in octaploids with higher con-
centration of secondary metabolites. Microscopic characters widely
used for the identification of ploidy levels viz pollen grain diameter
and stomatal length were also analyzed. The average stomatal size
was more in dodecaploid type. Pollen grains in the tetraploid type
were smaller (21.29 × 20.26 ␮m) as compared to other cytotypes
with the largest pollen grains seen in dodecaploid type measur-
ing 29.31 × 28.72 ␮m (Table 3). There seems a positive correlation
between the pollen and stomatal size with the level of ploidy. These
observations were in congruence with earlier reports (Inceer and
Ayaz, 2010; Pan, 1994). Polyploidization is an important process in
the evolution of most of the eukaryotic species. It often causes large
scale genomic reorganizations and is accompanied by a wide vari-
ety of phenotypic alterations in morphology, niche preference and
fitness characteristics (Riddle et al., 2006). The evolutionary suc-
cess of polyploids has often been attributed to the consequences
of having multiple genomes, with individuals of higher ploidy lev-
els considered to be more adaptable to differing conditions due to
genetic advantages that facilitate their establishment and persis-
tence (Comai, 2005). Change in environmental factors could change
the active principle present in the plants (Southwell and Bourke,
2001). In this relation, differences in longitude, latitude, humidity,
soil and temperature have pronounced impact on the production of
secondary metabolites. The metabolite profile of a plant defines its
ecological role and is therefore an important factor in understand-
ing the influence of environmental variables on the synthesis and
accumulation of secondary compounds.
The present investigation thus identifies existence of four novel
chromosomal races with chromosome numbers 2n = 40, 60, 80
and 120, wherein ploidy level range from tetraploidy to dode-
caploidy. These polyploid races are hitherto unreported in R.
nepalensis except for 2n = 120. Chemoprofiling based on four key
anthraquinones of different cytotypes from various altitudinal
gradients presented an appreciable variability in chrysophanol,
emodin and their glycoside contents. The increasing pattern of
metabolite accumulation broadly seems to be in conformity with
the increasing altitude and ploidy status. The species presents
robust adaptability and ecological plasticity possibly because of
the cytological variation. This study has a prospect to explore
desirable populations with higher content of chemical constituents
for commercial and pharmacological utilization. Many studies
considering multiple factors can only provide a thorough under-
standing of the pattern, establishment, survival and consequences
of higher ploidy levels across lineages under varying environmental
pressures.

Table 3
Morphological comparison of different cytotypes in Rumex nepalensis.

S. No. Characters Tetraploid Hexaploid Octaploid cytotype Octaploid cytotype Dodecaploid

cytotype 2n = 40 cytotype 2n = 60 (Y) 2n = 80 (T) 2n = 80 cytotype 2n = 120

1 Plant height (cm) 120–150 100–130 80–100 85–103 40–80

2 Branches/plant 7–10 4–7 10–15 9–13 6–9
3 Leaves/branch 12–15 3–7 4–7 5–8 4–6
4 Ochrea length 0.5–1 1–1.5 1–2 1–2 1–2
(mm)
5 Petiole length 2–2.5 1–1.5 1.5–2 1–2 1–5
(cm)
6 Leaf size (cm) L/B 2.5–4/1.5–2 3–5/2–3 5–6/3–3.5 4–6/3.5–4 7–10/4.5–6
7 Leaf surface Smooth Smooth Hairy Hairy Profusely hairy
8 Inflorescence 1–2 1.15–2.5 2–3 2–4 4–5
length (cm)
9 Average stomatal 19.68 × 8.78 20.01 × 9.08 23.01 × 10.91 23.13 × 11.02 25.98 × 12.68
size upper/lower
(␮m)
21.95 × 8.50 22.03 × 8.75 22.79 × 9.35 21.88 × 9.24 25.67 × 12.73
10 Pollen size (␮m) 21.29 × 20.26 22.56 × 21.75 23.74 × 22.23 22.98 × 22.06 29.31 × 28.72
11 Habit (Herb) Tallest Tall Medium Medium Small sized
12 Habitat Grows on moist Found in close Occurs on moist Occurs on moist Occurs in higher
rich soils area of water slopes slopes altitudes
streams
 U. Farooq et al. / Industrial Crops and Products 50 (2013) 112–117
Acknowledgments
The authors are grateful to the University grants commission
(UGC), New Delhi, for providing financial assistance under the DRS
SAP III programme and to the Department of Science and Technol-
ogy, New Delhi, for funding FIST programmes. One of the authors,
S.A.P is highly grateful to UGC for junior research fellowship. The
authors also thank Dr. S. K. Srivastava, Director Botanical Survey
of India (BSI), Northern circle (Dehradun) for his help in identi-
fication of plants. Thanks are also due to Dr Ram Vishwakarma,
Director IIIM Jammu, for providing necessary facilities for chemical
analysis. This manuscript represents institutional communication
number IIIM/1508/2012.
References
Aggarwal, P.K., Kumar, L., Garg, S.K., Mathur, V.S., 1986. Effect of Rumex nepalensis
extracts on histamine, acetylcholine, carbachol, bradykinin, and PGs evoked skin
reactions in rabbits. Annals of Allergy 56, 177–182.
Agnihotri, V.K., Lattoo, S.K., Thappa, R.K., Kaul, P., Qazi, G.N., Dhar, A.K., Saraf,
A., Kapahi, B.K., Saxena, R.K., Agarwal, S.G., 2005. Chemical variability in the
essential oil components of Achillea millefolium agg. from different Himalayan
habitats (India). Planta Medica 71, 280–283.
Carey, D.B., Wink, M., 1994. Elevational variation of quinolizidine alkaloid contents
in a lupine (Lupinus argenteus) of the Rocky Mountains. Journal of Chemical
Ecology 20, 849–857.
Comai, L., 2005. Advantages and disadvantages of polyploidy. Nature Reviews Genet-
ics 6, 836–846.
Darlington, C.D., Wylie, A.P., 1995. Chromosome Atlas of Flowering Plants. George
Allen and Unwin Ltd, London.
Degraeve, N., 1975. Contribution a l’etude cytotaxonomique des Rumex–I. Le genere
Rumex L. sensu stricto. Caryologia 28, 187–201.
Gachon, C.M.M., Meurinne, M.L., Saindrenan, P., 2005. Plant secondary metabolism
glycosyltransferases: the emerging functional analysis. Trends in Plant Science
10 (11), 542–549.
Gautam, R., Karkhile, K.V., Bhutani, K.K., Jachak, S.M., 2010. Anti-inflammatory,
cyclooxygenase (COX)-2, COX-1 inhibitory and free radical scavenging effects
of Rumex nepalensis. Planta Medica 76, 1564–1569.
Ghosh, L., Gayen, J.R., Sinha, S., Pal, S., Pal, M., Saha, B.P., 2003. Antibacterial efficacy
of Rumex nepalensis Spreng roots. Phytotherapy Research 17, 558–559.
Holmgren and Holmgren, 1998, http://sweetgum.nybg.org/ih/
Hussain, F., Ahmad, B., Hameed, I., Dastagir, G., Sanaullah, P., Azam, S., 2010. Antibac-
terial, antifungal and insecticidal activities of some selected medicinal plants of
Polygonaceae. African Journal of Biotechnology 9, 5032–5036.
Inceer, H., Ayaz, S.H., 2010. Chromosome numbers in Tripleurospermum Sch. Bip.
(Asteraceae) and closely related genera: relationships between ploidy level and
stomatal length. Plant Systematics and Evolution 285, 149–157.
Kaur, P., Chaudharya, A., Singh, R.D., Gopichand, P.R., Singh, B., 2012. Spatial and
temporal variation of secondary metabolite profiles in Ginkgo biloba leaves.
Chemistry and Biodiversity 9 (2), 409–417.
117
Khatoon, S., Ali, S.I., 1993. Chromosome Atlas of the Angiosperms of Pakistan. BCC &
T press, University of Karachi, Karachi.
Kirtikar, K.R., Basu, B.D., 1987. Indian Medicinal Plants, vol. 3. International Book
Distributors, India, pp. 2113–2114.
Kooke, R., Keurentjes, J.J.B., 2012. Multi-dimensional regulation of metabolic
networks shaping plant development and performance. Journal of Experimental
Botany 63 (9), 3353–3365.
Kumar, V., Subramanian, B., 1986. Chromosome Atlas of Flowering Plants of the
Indian Subcontinent, vol. 1. Dicotyledon, BSI, Calcutta.
Lattoo, S.K., Khan, S., Bamotra, S., Dhar, A.K., 2006a. Cytomixis impairs meio-
sis and influences reproductive success in Chlorophytum comosum (Thunb)
Jacq.—an additional strategy and possible implications. Journal of Bioscience 31,
629–637.
Lattoo, S.K., Dhar, R.S., Dhar, A.K., Sharma, P.R., Agarwal, S.G., 2006b. Dynamics of
essential oil biosynthesis in relation to inflorescence and glandular ontogeny in
Salvia sclarea. Flavour and Fragrance Journal 21 (5), 817–821.
Lavania, U.C., Srivastava, S., Lavania, S., Basu, S., Misra, N.K., Mukai, Y., 2012.
Autopolyploidy differentially influences body size in plants, but facilitates
enhanced accumulation of secondary metabolites, causing increased cytosine
methylation. Plant Journal 71, 539–549.
Liang, H.X., Dai, H.Q., Fu, H.A., Dong, X.P., Adebayo, A.H., Zhang, L.X., Cheng, Y.X.,
2010. Bioactive compounds from Rumex plants. Phytochemistry Letters 3,
181–184.
Manandhar, N.P., 2002. Plants and People of Nepal. Timber Press, Oregon, ISBN 0-
8192-527-6.
Moore, R.J., 1967–74. Index to plant chromosome numbers. Regnum Vegetabile
90,91,96.
Pan, J.T., 1994. Phylogeny classification and geographic distribution of Rodgersia
Gray. Acta Phytotaxonomica Sinica 92, 316–327.
Rana, S., Dhar, N., Bhat, W.W., Razdan, S., Khan, S., Dhar, R.S., Dutt, P., Lattoo,
S.K., 2012. A 12-deoxywithastramonolide-rich somaclonal variant in Witha-
nia somnifera (L.) Dunal—molecular cytogenetic analysis and significance as a
chemotypic resource. In Vitro Cellular and Developmental Biology—Plant 48 (5),
546–554.
Riddle, N.C., Kato, A., Birchler, J.A., 2006. Genetic variation for the response to ploidy
change in Zea mays L. Theoretical and Applied Genetics 114, 101–111.
Singh, N.P., Gupta, A.P., Sinha, A.K., Ahuja, P.S., 2005. High-performance thin
layer chromatography method for quantitative determination of four major
anthraquinone derivatives in Rheum emodi. Journal of Chromatography A 1077,
202–206.
Southwell, I.A., Bourke, A.C., 2001. Seasonal variation in hypericin content of Hyper-
icum perforatum L. (St. John’s wort). Photochemistry 56, 437–441.
Venkatesh, S., Reddy, B.M., Reddy, R.D., Ramesh, M., 2003. Antipyretic activity of
Rumex nepalensis roots. Nigerian Journal of Natural Products and Medicine 7,
53–54.
Zarinkamar, F., Tajik, S., Soleimanpour, S., 2011. Effects of altitude on anatomy and
concentration of crocin, picrocrocin and safranal in Crocus sativus L. Australian
Journal of Crop Science 5, 831–838.
Zhang, G.Q., Zhao, H.P., Wang, Z.Y., Cheng, J.R., Tang, X.M., 2008. Recent advances
in the study of chemical constituents and bioactivity of Rumex L. World Science
Technology (Modern Trading in China Medicine) 10, 86–93.
Zidorn, C., 2010. Altitudinal variation of secondary metabolites in flowering
heads of the Asteraceae: trends and causes. Phytochemistry Review 9,
197–203.