Chromosome Numbers and Genome Size Variation in Indian Species of Curcuma (Zingiberaceae)

Annals of Botany 100: 505–526, 2007
doi:10.1093/aob/mcm144, available online at www.aob.oxfordjournals.org
Chromosome Numbers and Genome Size Variation in Indian Species of Curcuma

(Zingiberaceae)
J A N A L E O N G - Š KOR NI ČKOVÁ 1 ,4, *, OTAK AR Š ÍD A 2 , VL AS TA J A RO L Í MOVÁ 3 , M A M YI L S AB U 4 ,
TO M Á Š F É R 1 , PAV E L T R Á V NÍ ČE K 1 ,3 and JA N SU DA 1,3
Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

1
Department of Botany, Charles University in Prague, Benátská 2, Prague 2, CZ-128 01, Czech Republic, 2Department of
Botany, National Museum in Prague, Zámek 1, Průhonice, CZ-252 43, Czech Republic, 3Botanical Institute, Academy of
Sciences of the Czech Republic, Zámek 1, Průhonice, CZ- 252 43, Czech Republic and 4Department of Botany,
Calicut University P.O., 673635, Kerala, India
Received: 1 February 2007 Returned for revision: 5 March 2007 Accepted: 15 May 2007 Published electronically: 8 August 2007
† Background and Aims Genome size and chromosome numbers are important cytological characters that signifi-
cantly influence various organismal traits. However, geographical representation of these data is seriously unba-
lanced, with tropical and subtropical regions being largely neglected. In the present study, an investigation was
made of chromosomal and genome size variation in the majority of Curcuma species from the Indian subcontinent,
and an assessment was made of the value of these data for taxonomic purposes.
† Methods Genome size of 161 homogeneously cultivated plant samples classified into 51 taxonomic entities was
determined by propidium iodide flow cytometry. Chromosome numbers were counted in actively growing root tips
using conventional rapid squash techniques.
† Key Results Six different chromosome counts (2n ¼ 22, 42, 63, .70, 77 and 105) were found, the last two repre-
senting new generic records. The 2C-values varied from 1.66 pg in C. vamana to 4.76 pg in C. oligantha, represent-
ing a 2.87-fold range. Three groups of taxa with significantly different homoploid genome sizes (Cx-values) and
distinct geographical distribution were identified. Five species exhibited intraspecific variation in nuclear DNA
content, reaching up to 15.1 % in cultivated C. longa. Chromosome counts and genome sizes of three Curcuma-
like species (Hitchenia caulina, Kaempferia scaposa and Paracautleya bhatii) corresponded well with typical hex-
aploid (2n ¼ 6x ¼ 42) Curcuma spp.
† Conclusions The basic chromosome number in the majority of Indian taxa (belonging to subgenus Curcuma)
is x ¼ 7; published counts correspond to 6x, 9x, 11x, 12x and 15x ploidy levels. Only a few species-specific
C-values were found, but karyological and/or flow cytometric data may support taxonomic decisions in some
species alliances with morphological similarities. Close evolutionary relationships among some cytotypes are
suggested based on the similarity in homoploid genome sizes and geographical grouping. A new species combi-
nation, Curcuma scaposa (Nimmo) Škorničk. & M. Sabu, comb. nov., is proposed.
Key words: Chromosome number, Curcuma, cytology, DNA C-value, flow cytometry, genome size, India, intraspecific
variation, polyploidy, taxonomy.
IN TROD UCT IO N Several taxonomic and biological problems have hin-

dered satisfactory systematic treatment of the genus.
The genus Curcuma L. (Zingiberaceae) contains many taxa
Original descriptions of many Curcuma species are vague
of economic, medicinal, ornamental and cultural import-
and inaccurate, and type specimens are often lacking or
ance, turmeric (C. longa L.) probably being the best
fragmentary. Proper preservation of Curcuma specimens
known. It is found throughout south and south-east Asia
is extremely difficult, exacerbating the limited amount of
with a few species extending to China, Australia and the
type material, leading to ambiguous name assignment and
South Pacific. The highest diversity is concentrated in
usage. In addition, high intra- and interpopulation variation
India and Thailand, with at least 40 species in each area,
has led to debate concerning species concepts and bound-
followed by Burma, Bangladesh, Indonesia and Vietnam.
aries. As a result, one species has often been described
Due to the lack of a comprehensive taxonomic revision,
repeatedly under different names whereas the same name
there is still little consensus on the number of species that
has been applied to different taxonomic entities.
should be recognized. Recent estimates vary from about
Some species may hybridize in the wild and the crosses
50 (Smith, 1981) to 80 (Larsen et al., 1998) and 100
may become naturalized (Škorničková and Sabu, 2005b;
species (Sirirugsa, 1996), although Škorničková et al.
Škorničková et al., 2007). Frequent cultivation of
(2004) suggested that their number will probably reach
Curcuma spp. and targeted selection of peculiar morpho-
120 in the near future in connection with detailed botanical
types have further contributed to taxonomic complexity of
exploration of India and south-east Asia.
the group. Moreover, polyploidy has played a significant
* For correspondence. Current address: The Herbarium, Singapore
role in evolution and diversification of various members
Botanic Gardens, Cluny Road 1, 259569, Singapore. of Zingiberaceae (e.g. Mukherjee, 1970; Lim, 1972a,b;
E-mail jana_skornickova@seznam.cz Poulsen, 1993; Chen and Chen, 1984; Takano, 2001;
# The Author 2007. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved.
For Permissions, please email: journals.permissions@oxfordjournals.org
506 Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma
Takano and Okada, 2002), including Curcuma (Prana et al., The infrageneric classification (subgenera Curcuma and
1978; Apavatjrut et al., 1996; Joseph et al., 1999; Ardiyani, Hitcheniopsis) generally followed the treatment of
2002; Sirisawad et al., 2003). It is well documented, in both Schumann (1904), with some modifications. In particular,
plants and animals, that an increase in ploidy level is com- C. petiolata and C. roscoeana were included in subgenus
monly associated with blurring of morphological bound- Curcuma based on the presence of two floral epigynous
aries between taxa (see Stace, 2000). glands (derived from gynopleural nectaries). This hitherto
The occurrence of different ploidy levels in Curcuma was neglected character (well developed in subgenus Curcuma
highlighted in early cytological studies (e.g. Suguira, 1931, but absent in subgenus Hitcheniopsis) seems to be pivotal
1936; Raghavan and Venkatasubban, 1943; Venkatasubban, for the updated subgeneric delimitation, better reflecting

1946; Chakravorti, 1948; Sharma and Bhattacharya, 1959), the current state of knowledge than did previous taxonomic
and 20 different somatic chromosome numbers have been concepts (J. Leong-Škorničková et al., unpubl. res.).
reported (Table 1). Considering the widely accepted basic
chromosome number x ¼ 21 (Ramachandran, 1961; Prana,
1977; Islam, 2004), this chromosomal variation roughly MATE RIA L AN D M ET HO DS
corresponds to three euploid levels (2x, 3x and 4x) plus Plant materials
several aneuploids. However, many of the records should
be treated with caution due to potential errors in chromo- In total, 161 individual plants belonging to 51 taxonomic
some counting as well as taxonomic ambiguity of the ana- entities were included in the study, 46 Curcuma species
lysed material, which is rarely documented by herbarium (31 assigned to species, 15 determined only tentatively
vouchers. Such a high basic chromosome number is likely or undetermined), one natural hybrid and four species
to be secondary, further complicating accurate inference often classified into separate but related genera: Hitchenia
of ploidy level and its evolution within the genus. caulina, Kaempferia scaposa, Paracautleya bhatii and
In addition to chromosome numbers and ploidy levels, Stahlianthus involucratus. The number of individuals per
genome size (nuclear DNA content) data also provide species varied from one to 16. Multiple samples were avail-
information useful in various fields of plant biology, includ- able for 26 species, whereas 25 taxa (12 Curcuma species,
ing systematics, evolution and conservation (Bennett and eight undetermined taxa, four species from related genera
Leitch, 2005b). In plants holoploid genome sizes (or and one hybrid) were represented by a single plant
1C-values) vary strikingly, ranging from about 0.065 to accession.
127.4 pg. Genome size variation has significant conse- Owing to difficulties surrounding systematic treatment of
quences at cellular, tissue and organismal levels and also the genus, specific names were assigned only to plant
influences phenological and ecological behaviour. Despite samples perfectly matching the species description. The
its usefulness in understanding plant evolution and diversifi- remaining samples were left unnamed in order to avoid mis-
cation, genome size variation in Curcuma is not well docu- leading information resulting from unambiguous identifi-
mented, and estimates for only a few species have been cation. All plants were collected in the wild on the Indian
published (Table 1). Bharathan et al. (1994) determined by peninsula, often at or near the locus classicus, during
flow cytometry 1C ¼ 1.30 pg in C. zanthorrhiza and Das 2000– 2004 (Table 2) and are being grown at the Calicut
et al. (1999) used cytophotometry to study genome size in University Botanical Garden, Kerala, India (11 8350 N, 70
C. amada (4C ¼ 3.120 pg), C. caesia (4C ¼ 4.234 pg) and 8450 E, 50 m a.s.l.). Geographical positions of the collection
C. longa (4C ¼ 5.100 – 5.263 pg). Curcuma longa was also localities are shown in Fig. 1. Herbarium vouchers are
investigated by Nayak et al. (2006), who observed deposited in CALI, with duplicates in MH and SING;
4C-values ranging from 4.30 to 8.84 pg in 17 varieties. incomplete sets are also kept in CAL, K and PR; vouchers
In addition, flow-cytometric nuclear DNA amounts for of C. oligantha from Sri Lanka are deposited in PDA and
16 taxa (including the above-mentioned species and one SING. In addition, a large collection of photographic docu-
undetermined sample) from Bangladesh are given in the mentation of living material (including details of flower
unpublished PhD thesis of Islam (2004). morphology) is available for each accession (see Fig. 5).
As a part of ongoing comprehensive taxonomic revision
of Curcuma in India (see Škorničková et al., 2003a,b,
Genome size estimation
2004, 2007; Škorničková and Sabu, 2005a,b,c), the
present study aimed to provide a detailed survey of chromo- Nuclear DNA C-values ( ¼ holoploid genome sizes) and
somal and genome size variation in the majority of known Cx-values ( ¼ monoploid genome sizes) were estimated
Indian species. In particular, we address whether genome using propidium iodide flow cytometry (FCM). Sample
size and chromosome numbers can be used as taxonomi- preparation generally followed the two-step procedure orig-
cally informative markers for species delimitation and inally described by Otto (1990). Plants were cultivated for
whether they can elucidate the taxonomic position of four at least 1 year under homogeneous conditions. About
species with Curcuma-like morphological traits often 1 cm2 of young and intact fresh leaf tissue and internal stan-
placed in separate genera (Hitchenia caulina, Kaempferia dard was co-chopped in a sandwich-like arrangement with a
scaposa, Paracautleya bhatii and Stahlianthus involucra- sharp razor blade in 1 mL of ice-cold Otto I buffer (0.1 M
tus). The issue of basic chromosome number and the citric acid, 0.5 % Tween 20). The nuclear suspension was
origin of the polyploid taxa are also discussed in the light filtered through a nylon mesh (42-mm pore size) and centri-
of the findings. fuged at 150g for 5 min. The supernatant was then removed
TA B L E 1. A synopsis of published chromosome counts and genome sizes in the genus Curcuma
No. of chromosomes

Genome Origin of
Species n 2n size (pg)* plant material Reference
subgen. Curcuma K.Schum.

C. aeruginosa Roxb. 63 Indonesia Prana (1977, 1978)
Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma

63 Thailand Apavatjrut et al. (1996)
63 India Joseph et al. (1999)
63 Thailand Paisooksantivatana and Thepsen (2001)
63 Indonesia Ardiyani (2002)
28– 35 63 Thailand Sirisawad et al. (2003)
63, 84 3.203– 5.302 Bangladesh Islam (2004)
C. amada Roxb 42 India Raghavan and Venkat. (1943)
42 India Chakravorti (1948)
42 India Raghavan and Arora (1958)
42 India Sharma and Bhattacharya (1959)
42 India Ramachandran (1961, 1969)
40 4.234/4C India Das et al. (1999)
42 2.132 Bangladesh Islam (2004)
C. amarissima Roscoe 63 3.289 Bangladesh Islam (2004)
C. angustifolia Roxb. 42 India Chakravorti (1948)
42 India Sharma and Bhattacharya (1959)
42 2.121– 2.141 Bangladesh Islam (2004)
C. aromatica Salisb. 42 India Raghavan and Venkat (1943)
42 India Chakravorti (1948)
63, 86 India Ramachandran (1961, 1969)
84 India Nambiar et al. (1982)
63 China Chen and Chen (1984)
42 India Sarkar (1990)
C. attenuata Wall. ex Baker 42 84 Thailand Apavatjrut et al. (1996)
42 84 Thailand Sirisawad et al. (2003)
C. australasica Hook.f 2.153– 1.181 Bangladesh Islam (2004)
C. aurantiaca Zijp 42 Indonesia Prana (1977, 1978)
21 Pen. Malaysia Beltran and Kam (1984)
C. brog Valeton 63, 64, 70 Indonesia Prana (1977)
63, 64 Indonesia Prana (1978)
C. caesia Roxb. 22 3.12/4C India Das et al. (1999)
63 India Joseph et al. (1999)
C. colorata Valeton 62, 63 Indonesia Prana (1977, 1978)
C. comosa Craib 42 India Joseph et al. (1999)
C. cf. comosa Roxb. 63 Thailand Paisooksantivatana and Thepsen (2001)
C. decipiens Dalzell 21 42 India Ramachandran (1961, 1969)
C. elata Roxb. 63 Thailand Apavatjrut et al. (1996)
C. haritha Mangaly & M. Sabu 42 India Joseph et al. (1999)
507
Continued
TA B L E 1. Continued
508
No. of chromosomes

Genome Origin of
Species n 2n size (pg)* plant material Reference
C. heyneana Valeton & Zijp 63 Indonesia Prana (1977, 1978)

63 Indonesia Ardyiani (2002)
C. kwangsiensis S.G.Lee & C.F.Liang 42 84 China Chen et al. (1988)

C. latifolia Roscoe 63 3.435 Bangladesh Islam (2004)
C. longa L. 64 Unknown Sugiura (1931, 1936)
62 India Raghavan and Venkat (1943)
62, 63, 64 India Chakravorti (1948)
32 Unknown Sato (1948)
62, 93 India Sharma and Bhattacharya (1959)
63 Indonesia Prana (1977, 1978)
48 5.1 –5.26/4C India Das et al. (1999)
48 4.30–8.84/4C India Nayak et al. (2006)
C. malabarica Velay., Mural. & Amalraj 42 India Joseph et al. (1999)
C. mangga Valeton & Zijp 42 Indonesia Prana (1977, 1978)
63 Indonesia Ardyiani (2002)
C. neilgherrensis Wight 42 India Chakravorti (1948)
C. aff. oligantha Trimen 42 Thailand Eksomtramage et al. (2002)
40 Thailand Saensouk and Chantaranothai (2003)
C. petiolata Roxb. 64 India Venkatasubban (1946)
42 Indonesia Prana (1977, 1978)
C. cf. petiolata Roxb. 42 Thailand Paisooksantivatana and Thepsen (2001)
C. phaeocaulis Valeton 62, 63, 64 Indonesia Prana (1977, 1978)
C. purpurascens Blume 63 Indonesia Prana (1977, 1978)
C. raktakanta Mangaly & M.Sabu 63 India Joseph et al. (1999)
C. roscoeana Wall. 21 42 Thailand Apavatjrut et al. (1996)
42 Thailand Eksomtramage et al. (1996a,b)
C. rubescens Roxb. 28– 35 63 Thailand Sirisawad et al. (2003)
42 Bangladesh Islam (2004)
C. sessilis Gage 84 Thailand Saensouk et al. (1998)
46, 92 Thailand Saensouk and Chantaranothai (2003)
C. soloensis Valeton 63 Indonesia Prana (1977, 1978)
C. viridiflora Roxb. 42 2.164– 2.173 Bangladesh Islam (2004)
C. wenyujin Y.H.Chen & C.Ling 63 China Chen and Chen (1984)
C. zanthorrhiza Roxb. 63 Indonesia Prana (1977, 1978)
63 China Chen and Chen (1984)
1.30/1C Origin unknown Bharathan et al. (1994)
C. zedoaria (Christm.) Roscoe 64 India Venkatasubban (1946)
63, 64 India Chakravorti (1948)

66 India Sharma (1970)
63, 64, 66 Indonesia Prana (1977, 1978)
66 India Chatterjee et al. (1989)

subgen. Hitcheniopsis (Baker) K.Schum.
C. alismatifolia Gagnep. 16 32 Thailand Apavatjrut et al. (1996)
32 Thailand Saensouk et al. (1998)
C. gracillima Gagnep. 24 Thailand Saensouk et al. (1998)
C. cf. gracillima Gagnep. 40 Thailand Paisooksantivatana and Thepsen (2001)
C. harmandii Gagnep. 10 20 Thailand Eksomtramage et al. (1996a,b)
C. parviflora Wall. 14, 17, 18, 28 28, 34, 36 Thailand Apavatjrut et al. (1996)
32 Thailand Weerapakdee and Krasaechai (1997)
30 Thailand Saensouk et al. (1998)
32 Thailand Eksomtramage et al. (2002)
30 Thaialnd Saensouk and Chantaranothai (2003)
12, 14, 17, 18, 28 24, 28, 34, 36, 56 Thailand Sirisawad et al. (2003)
C. rhabdota Sirirugsa & M.F.Newman 24 Thailand Eksomtramage et al. (2002)
C. thorelii Gagnep. 36 Thailand Eksomtramage and Boontum (1995)
17 34 Thailand Apavajrut et al. (1996)
38? Thailand Ardiyani (2002)
* Genome sizes expressed in 2C-values unless indicated otherwise.
509
510
TA B L E 2. 2C nuclear DNA content with standard error, mean value for a species, intraspecific variation, 1C-value expressed in DNA picograms and megabase pairs

(1 pg ¼ 978 Mbp), somatic chromosome number (2n), ploidy level, homoploid genome size (Cx-value, determined as 2C DNA amount/ploidy level) and its mean for a
species, internal standard used, locality, species distribution pattern, and a field accession number for 161 Indian plants belonging to 51 taxa of Curcuma and related
genera.
Ploidy
level (x)

Mean 1C- 1C- or DNA Mean
2C-value 2C-value Intraspecific value value ploidy 1Cx-value Cx-value Internal Distribution
Group/species Field no. (pg)+s.e. (pg)* variation (%) (pg) (Mbp) 2n † level (pg) (pg)* standard‡ Locality pattern§
DIPLOID (x ¼ 11)
C. vamana M. Sabu & 84156 1.663+ 1.66u 0.83 813 22! 2 0.83 0.83b L Kerala, Trichur SW
Mangaly 0.008 Dt.
HEXAPLOIDS–GROUP I
C. amada Roxb. 71421 1.882+ 1.86st 3.6 0.94 920 6 0.31 0.31lmnopqrs G W. Bengal, NE & E
0.002 Kolkata
71472 1.854+ 0.93 907 42 6 0.31 L W. Bengal,
0.007 Darjeeling Dt.
73440 1.816+ 0.91 888 6 0.30 L Bihar, Bhagalpur
0.002 Dt.
73482 1.873+ 0.94 916 6 0.31 G W. Bengal,
0.004 Kolkata
73484 1.875+ 0.94 917 6 0.31 G W. Bengal,
0.006 Kolkata
st lmnopqrs
C. aromatica Salisb. 73423 1.900+ 1.86 3.5 0.95 929 42 6 0.32 0.31 L Sri Lanka, Kegalle SW & SL
s.l.– sp. 1 0.008 Dt.
84109 1.846+ 0.92 903 42 6 0.31 L Kerala, Kollam
0.006 Dt.
84114 1.880+ 0.94 919 6 0.31 G Kerala, Kollam
0.006 Dt.
84123 1.876+ 0.94 917 6 0.31 G Kerala, Wynad Dt.
0.001
84123-II 1.881+ 0.94 920 6 0.31 G Kerala, Wynad Dt.
0.004
84170 1.865+ 0.93 912 6 0.31 G Kerala, Idukki Dt.
0.006
0.001
84183A 1.836+ 0.92 898 6 0.31 G Kerala, Idukki Dt.
0.009
84183B 1.838+ 0.92 899 6 0.31 G Kerala, Idukki Dt.
0.002
C. mangga Valeton & Zijp 84101 1.807+ 1.83t 3.2 0.90 884 6 0.30 0.31mnopqrs G Kerala, Ernakulam SW
0.001 Dt.
84115 1.862+ 0.93 911 6 0.31 L Kerala, Kollam
0.008 Dt.
84149 1.851+ 0.93 905 6 0.31 G Kerala, Trichur
0.002 Dt.
84150 1.804+ 0.90 882 42 6 0.30 G Kerala, Trichur
0.002 Dt.
C. montana Roxb. 71484 1.816+ 1.79tu 6.1 0.91 888 42! 6 0.30 0.30rs L Jharkhand, Ranchi E&C
0.006 Dt.
73419 1.854+ 0.93 907 42! 6 0.31 L Jharkhand,
0.008 Paschim

Singhbum Dt.
73425 1.823+ 0.91 891 6 0.30 G Orissa, Koraput
0.004 Dt.
73430 1.786+ 0.89 873 6 0.30 L Orissa, Koraput
0.008 Dt.
73433 1.806+ 0.90 883 6 0.30 G Jharkhand,

0.004 Paschim
Singhbum Dt.
73433-II 1.778+ 0.89 869 6 0.30 G Jharkhand,
0.002 Paschim
Singhbum Dt.
73437 1.774+ 0.89 867 6 0.30 G Jharkhand,
0.001 Paschim
Singhbum Dt
73456 1.763+ 0.88 862 6 0.29 G Jharkhand, Ranchi
0.004 Dt.
73471 1.767+ 0.88 864 6 0.29 G Chhattisgarh,
0.004 Jagdalpur Dt.
73473 1.782+ 0.89 871 6 0.30 G Chhattisgarh,
0.004 Jagdalpur Dt.
73474 1.774+ 0.89 867 6 0.30 G Chhattisgarh,
0.008 Jagdalpur Dt.
73479 1.748+ 0.87 855 6 0.29 G Chhattisgarh,
0.002 Bilaspur Dt.
st klmnopqr
C. prakasha S. Tripathi 71441 1.877+ 1.87 4.5 0.94 918 ca 6 0.31 0.31 L Meghalaya, NE
0.003 42! Ribhoi Dt.
71442 1.847+ 0.92 903 6 0.31 L Meghalaya,
0.005 Ribhoi Dt.
71450 1.918+ 0.96 938 6 0.32 G Meghalaya,
0.009 S. Garo Hills Dt.
71462 1.893+ 0.95 926 42! 6 0.32 G Meghalaya,
0.002 E. Garo Hills Dt.
71463 1.835+ 0.92 897 6 0.31 G Meghalaya,
73406 1.872+ 0.94 915 6 0.31 G Meghalaya,
0.001 E. Khasi Hills Dt.
s k
C. roscoeana Wall. 73309 1.962+ 1.96 0.98 959 42 6 0.33 0.33 G Andaman ANI
0.005 Isls. M. Andaman
C. rubescens Roxb. 71454 1.889+ 1.87st 2.5 0.94 924 6 0.31 0.31lmnopqrs L Meghalaya, NE
71457 1.843+ 0.92 901 ca 42 6 0.31 L Meghalaya,
C. rubrobracteata Škorničk., 86241 1.836+ 1.84st 0.92 898 42! 6 0.31 0.31mnopqrs L Mizoram, NE
M. Sabu & Prasanthk. 0.008 Lawngtlai Dt.
st lmnopqrs
C. sp. ‘sulphurea’ 86231 1.853+ 1.85 0.93 906 42! 6 0.31 0.31 G Mizoram, Lunglei NE
0.002 Dt.
C. sp. ‘repens’ 71456 1.834+ 1.83st 0.92 897 42! 6 0.31 0.31mnopqrs L Meghalaya, NE
Continued
511
512
Ploidy

level (x)
†
Group/species Field no. (pg)+s.e. (pg)* variation (%) (pg) (Mbp) 2n level (pg) (pg)* standard‡ Locality pattern§
HEXAPLOIDS–GROUP II

C. angustifolia Roxb. 73449 2.148+ 2.15r 2.3 1.07 1050 ca 42 6 0.36 0.36j G Uttaranchal, Dehra N&E
0.003 Dun Dt.
73452 2.158+ 1.08 1055 42 6 0.36 G Uttaranchal, Tehri
0.007 Gharwal Dt.
73452-II 2.162+ 1.08 1057 6 0.36 G Uttaranchal, Tehri
0.007 Gharwal Dt.
73454 2.145+ 1.07 1049 6 0.36 G Uttaranchal, Tehri
0.001 Gharwal Dt.
73465 2.114+ 1.06 1034 6 0.35 G Jharkhand,
0.004 Sahibganj Dt.
73480 2.153+ 1.08 1053 6 0.36 G Chhattisgarh,
0.006 Bilaspur Dt.
C. aurantiaca Zijp 73455 2.223+ 2.20qr 3.1 1.11 1087 6 0.37 0.37hij G Kerala, Kozhikode SW
0.008 Dt.
84108 2.156+ 1.08 1054 42 6 0.36 G Kerala, Kollam
0.007 Dt.
77019 2.206+ 1.10 1079 42 6 0.37 G Kerala, Kozhikode
0.004 Dt.
op efg
C. cannanorensis R. Ansari, 84144 2.330+ 2.33 0.0 1.17 1139 42! 6 0.39 0.39 G Kerala, Kannur SW
V. J. Nair & N. C. Nair 0.007 Dt.
84164 2.331+ 1.17 1140 42! 6 0.39 B Karnataka, Udupi
0.005 Dt.
op e
C. decipiens Dalzell 73445 2.363+ 2.35 2.8 1.18 1156 6 0.39 0.39 B Maharashtra, W
0.010 Sindudurg Dt.
84179 2.305+ 1.15 1127 6 0.38 G Maharashtra,
0.003 Sindudurg Dt.
84179 2.370+ 1.19 1159 42 6 0.40 B Maharashtra,
0.007 Sindudurg Dt.
C. inodora Blatt. 73403 2.290+ 2.29pq 1.15 1120 42! 6 0.38 0.38efg G Maharashtra, W
0.006 Thane Dt.
op ef
C. karnatakensis Amalraj, 84163 2.340+ 2.34 1.17 1144 42! 6 0.39 0.39 B Karnataka, Uttar SW
Velay. & Mural 0.012 Kannad Dt.
C. kudagensis Velay., 84152 2.287+ 2.29pq 1.14 1118 42! 6 0.38 0.38efgh B Karnataka, SW
V. S. Pillai & Amalraj 0.007 Kodagu Dt.
pq efg
C. neilgherrensis Wt. 73490 2.251+ 2.29 3.8 1.13 1101 6 0.38 0.38 G Tamil Nadu, S & SW
0.011 Nilgiris Dt.
84157 2.336+ 1.17 1142 6 0.39 G Kerala, Wyanad
0.001 Dt.
84174 2.297+ 1.15 1123 42 6 0.38 B Tamil Nadu,
0.005 Nilgiris Dt.
84181 2.273+ 1.14 1111 6 0.38 G Kerala, Wyanad
0.001 Dt.
pqr fghi
C. pseudomontana J. Graham 73401 2.225+ 2.25 2.4 1.11 1088 6 0.37 0.38 G Maharashtra, Pune W
0.005 Dt.
73402 2.279+ 1.14 1114 42! 6 0.38 B Maharashtra, Pune
0.010 Dt.
C. reclinata Roxb. 73477 2.313+ 2.29pq 3.9 1.16 1131 42! 6 0.39 0.38efgh G Chhattisgarh, C
0.002 Bilaspur Dt.
73467 2.226+ 1.11 1089 6 0.37 G Madhya Pradesh,
0.008 Hoshangabad Dt.

73469 2.292+ 1.15 1121 6 0.38 G Madhya Pradesh,
0.003 Satna Dt.
73469 2.309+ 1.15 1129 6 0.38 G Madhya Pradesh,
0.007 Satna Dt.
HEXAPLOIDS–GROUP III
C. coriacea Mangaly & 73447 2.603+ 2.60lm 1.30 1273 42! 6 0.43 0.43c B Kerala, Idukki Dt. SW

M. Sabu 0.006
mn d
C. mutabilis Škorničk., 84145 2.492+ 2.49 1.25 1219 42! 6 0.42 0.42 B Kerala, SW
M. Sabu & Prasanthk. 0.004 Malappuram Dt.
C. sp. ‘aff. prakasha’ 71443 2.446+ 2.45no 1.22 1196 42! 6 0.41 0.41d B Meghalaya, NE
0.005 Ribhoi Dt.
mn d
C. sp. 73417 2.501+ 2.50 1.25 1223 42! 6 0.42 0.42 L Assam, NE
0.001 Bongaigaon Dt.
NONAPLOIDS
C. aeruginosa Roxb. 71431 2.825+ 2.86efg 3.2 1.41 1381 63 9 0.31 0.32klmn L Assam, S
84119 2.808+ 1.40 1373 9 0.31 L Kerala, Ernakulam
0.012 Dt.
84130 2.898+ 1.45 1417 9 0.32 G Kerala, Kozhikode
0.003 Dt.
86102 2.876+ 1.44 1406 9 0.32 G Kerala, Kottayam
0.005 Dt.
86354 2.889+ 1.44 1413 9 0.32 G Andaman Isls.,
0.005 S. Andaman
efghi klmnopq
C. aromatica Salisb. 71460 2.863+ 2.83 1.7 1.43 1400 9 0.32 0.31 L Meghalaya, NE
s.l.– sp. 2 0.014 E. Garo Hills Dt.
73410 2.844+ 1.42 1391 9 0.32 G Meghalaya,
71445 2.815+ 1.41 1377 9 0.31 L Meghalaya,
71447 2.822+ 1.41 1380 63 9 0.31 L Meghalaya,
71453 2.820+ 1.41 1379 9 0.31 L Meghalaya,
C. aromatica Salisb. 71486 2.694+ 2.68jkl 0.9 1.35 1317 9 0.30 0.30rs G Jharkhand, E
s.l.– sp. 3 0.007 Devghar Dt.
71488 2.686+ 1.34 1313 9 0.30 G Jharkhand, Dumka
0.002 Dt.
0.007 Dt.
0.012 Dt.
C. caesia Roxb. 71418 2.825+ 2.82efghi 4.9 1.41 1381 9 0.31 0.31klmnopq G Unknown, NE & E
0.011 cultivated at
CUBG
71439 2.823+ 1.41 1380 ca 63 9 0.31 G Meghalaya,
0.016 Ribhoi Dt.
71439A 2.830+ 1.42 1384 9 0.31 L Meghalaya,
0.011 Ribhoi Dt.
513
Continued
514
Ploidy

level (x)
†
Group/species Field no. (pg)+s.e. (pg)* variation (%) (pg) (Mbp) 2n level (pg) (pg)* standard‡ Locality pattern§
71439B 2.782+ 1.39 1360 9 0.31 L Meghalaya,

0.007 Ribhoi Dt.
71451 2.893+ 1.45 1415 9 0.32 G Meghalaya,
71459 2.892+ 1.45 1414 9 0.32 L Meghalaya,
71469 2.759+ 1.38 1349 9 0.31 L W. Bengal,
71490 2.825+ 1.41 1381 9 0.31 G Jharkhand, Pakur
0.004 Dt.
86222 2.788+ 1.39 1363 9 0.31 G Mizoram, Lunglei
0.011 Dt.
efgh klmnop
C. codonantha Škorničk., 73319 2.838+ 2.84 1.42 1388 63! 9 0.32 0.32 G Andaman Isls., ANI
M. Sabu & Prasanthk. 0.010 N. Andaman
C. elata complex
C. elata Roxb. 86321 2.853+ 2.85efg 1.43 1395 9 0.32 0.32klmno L Andaman Isls., ANI
0.014 S. Andaman
C. latifolia Roscoe 73321 2.801+ 2.80efghij 1.40 1370 9 0.31 0.31lmnopqrs G Andaman Isls., ANI
0.005 M. Andaman
e kl
C. sp. ‘elata-latifolia’ 71419 2.853+ 2.91 3.1 1.43 1395 9 0.32 0.32 L Uttaranchal, Dehra N, E & NE
0.010 Dun Dt.
71423 2.941+ 1.47 1438 63 9 0.33 L W. Bengal,
0.012 Jalpaiguri Dt.
71440 2.905+ 1.45 1421 9 0.32 G Meghalaya,
0.006 Ribhoi Dt.
71448 2.889+ 1.44 1413 9 0.32 L Meghalaya,
71461 2.884+ 1.44 1410 9 0.32 G Meghalaya,
71471 2.925+ 1.46 1430 9 0.33 L W. Bengal,
71477 2.940+ 1.47 1438 9 0.33 G W. Bengal,
71483 2.930+ 1.47 1433 9 0.33 G Uttaranchal, Dehra
0.008 Dun Dt.
73412 2.942+ 1.47 1439 9 0.33 G W. Bengal,
0.005 Jalpaiguri Dt.
C. ferruginea Roxb. 71479 2.805+ 2.80efghijk 2.5 1.40 1372 9 0.31 0.31lmnopqrs G W. Bengal, E & ANI
0.005 S. 24-Prghanas Dt.
73320 2.814+ 1.41 1376 9 0.31 G Andaman Isls.,
0.001 N. Andaman
73320-II 2.818+ 1.41 1378 9 0.31 G Andaman Isls.,
0.008 N. Andaman
86334B 2.749+ 1.37 1344 9 0.31 G Andaman Isls.,
0.006 M. Andaman
C. leucorhiza Roxb. 71489 2.714+ 2.70jkl 1.0 1.36 1327 9 0.30 0.30qrs L Jharkhand, Dumka E
0.013 Dt.
71493 2.688+ 1.34 1314 9 0.30 G Jharkhand, Pakur
0.005 Dt.

73441 2.692+ 1.35 1316 63! 9 0.30 L Bihar, Bhagalpur
0.015 Dt.
C. longa L. 71420 2.621+ 2.71ijkl 15.1 1.31 1282 9 0.29 0.30pqrs L W. Bengal, S, C, E &
0.011 Kolkata NE
71422 2.580+ 1.29 1262 63 9 0.29 L W. Bengal,
0.007 Kolkata

71433 2.817+ 1.41 1378 63 9 0.31 L Assam,
71436 2.603+ 1.30 1273 9 0.29 L Assam, Dispur Dt.
0.012
71473 2.603+ 1.30 1273 9 0.29 L W. Bengal,
71480 2.653+ 1.33 1297 9 0.29 G W. Bengal,
0.004 S. 24-Parghanas
Dt.
71487 2.969+ 1.48 1452 9 0.33 G Jharkhand,
0.005 Devgar, Dt.
73303 2.751+ 1.38 1345 9 0.31 G Andaman Isls.,
0.004 N. Andaman
73411 2.715+ 1.36 1328 9 0.30 G Meghalaya,
0.002 Ribhoi Dt.
73478 2.766+ 1.38 1353 9 0.31 G Chhattisgarh,
0.001 Bilaspur Dt.
84127 2.677+ 1.34 1309 9 0.30 G Kerala, Wyanad
0.002 Dt.
84154 2.665+ 1.33 1303 9 0.30 G Kerala, Palghat
0.004 Dt.
84160 2.656+ 1.33 1299 9 0.30 G Kerala, Wyanad
0.007 Dt.
86221 2.841+ 1.42 1389 9 0.32 G Mizoram, Lunglei
0.006 Dt.
86221-II 2.846+ 1.42 1392 9 0.32 G Mizoram, Lunglei
0.006 Dt.
86221-III 2.829+ 1.41 1383 9 0.31 G Mizoram, Lunglei
0.004 Dt.
ef klm
C. zanthorrhiza Roxb. 73302 2.896+ 2.88 2.9 1.45 1416 9 0.32 0.32 G Andaman Isls., S & ANI
0.003 N. Andaman
84107 2.901+ 1.45 1419 9 0.32 G Kerala, Kollam
0.007 Dt.
0.003
84182 2.820+ 1.41 1379 63 9 0.31 L Kerala, Idukki Dt.
0.015
84182A 2.839+ 1.42 1388 9 0.32 G Kerala, Idukki Dt.
0.003
C. sp. ‘fucata’ 71430 2.805+ 2.80efghij 1.40 1372 63! 9 0.31 0.31lmnopqrs L Assam, NE
fghijk mnopqrs
C. sp. ‘man-and’ 86306 2.804+ 2.76 3.1 1.40 1371 63! 9 0.31 0.31 L Andaman Isls., ANI
0.006 S. Andaman
515
Continued
516

Ploidy
level (x)
Group/species Field no. (pg)+s.e. (pg)* variation (%) (pg) (Mbp) 2n † level (pg) (pg)* standard‡ Locality pattern§

86313 2.721+ 1.36 1331 9 0.30 G Andaman Isls.,
0.009 S. Andaman
efghijk lmnopqrs
C. sp. ‘picta’ 71452 2.803+ 2.81 3.7 1.40 1371 9 0.31 0.31 L Meghalaya, NE & SL
71464 2.782+ 1.39 1360 9 0.31 L Meghalaya,
71465 2.753+ 1.38 1346 9 0.31 L Assam, Kokrajar
0.009 Dt.
73422 2.854+ 1.43 1396 9 0.32 G Sri Lanka, Kegalle
0.004 Dt.
hijkl opqrs
C. sp. ‘roxburgh’ 71434 2.717+ 2.72 1.36 1329 ca 9 0.30 0.30 L Assam, NE
0.012 63! Bongaigaon Dt.
C. sp. ‘tikhur’ 73476 2.735+ 2.74ghijk 1.37 1337 9 0.30 0.30nopqrs G Chhattisgarh, C
0.001 Bilaspur Dt.
kl s
C. sp. ‘aff. zanthorrhiza’ 73420 2.672+ 2.67 0.5 1.34 1307 9 0.30 0.30 G Jharkhand, E
0.005 Paschim
Singhbum Dt
73420A 2.663+ 1.33 1302 9 0.30 G Jharkhand,
0.013 Paschim
Singhbum Dt
73420B 2.676+ 1.34 1309 9 0.30 G Jharkhand,
0.011 Paschim
Singhbum Dt
11-PLOID
C. oligantha Trimen 73325 4.755+ 4.76a 2.38 2325 77!! 11 0.43 0.43c B Sri Lanka, Badulla SL
0.026 Dt.
cf. 12-PLOID
C. sp. ‘ranchi’ 71485 3.713+ 3.71c 1.86 1816 . ca 12 0.31 0.31lmnopqrs G Jharkhand, Ranchi E
0.005 70! Dt.
15-PLOIDS
C. raktakanta Mangaly & 73414 4.614+ 4.57b 5.1 2.31 2256 15 0.31 0.30nopqrs G Assam, SW & NE
M. Sabu 0.010 Bongaigaon Dt.
71432 4.413+ 2.21 2158 105!! 15 0.29 L Assam,
84120 4.637+ 2.32 2267 15 0.31 L Kerala, Ernakulam
0.012 Dt.
84120 4.637+ 2.32 2267 105!! 15 0.31 G Kerala, Ernakulam
0.012 Dt.
84148 4.638+ 2.32 2268 15 0.31 G Kerala, Trichur
0.003 Dt.
HYBRID
C. angustifoliamontana 73480B 1.903+ 1.90st 0.95 931 6 0.32 0.32klmno G Chhattisgarh, C
0.005 Bilaspur Dt.
Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma 517
and nuclei were gently resuspended in 100 mL of fresh Otto I

buffer. After incubation (15 min at room temperature), 1 mL
SW
NE
W
W
of Otto II buffer (0.4 M Na2HPO4.12H2O) supplemented
with propidium iodide (at a final concentration 50 mL mL –
1
), RNase IIA (50 mL mL – 1) and 2-mercaptoethanol (2 mL
Karnataka, Udupi
mL – 1) was added. The samples were incubated for 30 min
B, Bellis perennis L. (2C ¼ 3.96 pg); G, Glycine mas ‘Polanka’ (2C ¼ 2.5 pg, primary reference standard); L, Solanum lycopersicum ‘Stupnické polnı́ tyčkové rané’ (2C ¼ 2.21 pg).
S. Garo Hills Dt.
Goa, N. Goa Dt.
at room temperature, after which fluorescence intensity of

Maharashtra,
Meghalaya,
Chromosome numbers determined in the present work, ! new species count, !! new count for a genus. Chromosome numbers taken from the literature are given in parentheses.
5000 particles was recorded on a Partec Cyflow instrument
Satara Dt.
(Partec GmbH, Münster, Germany) equipped with a 532-nm

solid-state laser (Cobolt Samba 100 mW, Cobolt, Sweden).
Dt.
Each plant was re-analysed at least three times on different

days and only histograms with peaks of approximately the
same height were accepted. If between-day variation (max./
G
G
B
min. value) exceeded 2 %, the outlying value was discarded

and the sample re-measured. Glycine max ‘Polanka’ (2C ¼
2.50 pg; Doležel et al., 1994) was selected as a primary
internal reference standard. Solanum lycopersicum
0.37ghi
0.39efg
1.56 a
0.36ij
L. ‘Stupické polnı́ tyčkové rané’ (2C ¼ 2.11 pg) and Bellis

perennis L. (2C ¼ 3.96 pg) were used as secondary reference
standards for Curcuma samples with low and high genome
0.37
0.39
0.36
1.56
ANI, Andaman Islands; C, Central India; E, Eastern India; N, Northern India; S, Southern India; SL, Sri Lanka; W, Western India.
sizes, respectively, in order to minimize standard-to-sample

peak ratio and thus avoid potential non-linearity of FCM
measurements. Genome sizes of secondary reference stan-
dards were calibrated against the primary one, based on nine
6
replications on different days. The total number of flow cyto-

metric measurements for Curcuma was 678.
(22)
42!
42!
42!
Chromosome counts
Chromosome numbers were counted in actively growing
1098
1141
1067
1523
root tips of the cultivated plants. Samples were pretreated

with a saturated solution of p-dichlorbenzene (3 h, room
1.12
1.17
1.09
1.56
temperature), fixed in a 3:1 mixture of ethanol and acetic

acid (4 h, 4 8C), macerated in 1:1 hydrochloric acid/ethanol
(30 s, room temperature) and immediately squashed in a
drop of lactopropionic orceine. The number of chromosomes
* Letters indicate group of taxa that are not significantly different at a ¼ 0.05.
was determined in 5 – 10 complete well-spread mitotic plates

using a Carl-Zeiss Jena NU microscope equipped with an
Olympus Camedia C-2000 Z camera.
2.24pqr
2.18qr
2.33op
3.11d
Statistical analysis
Statistical analyses were performed in the SAS 8.1 stat-
istical package (SAS Institute, Cary, NC, USA). Between-
2.245+
2.333+
2.181+
3.114+
0.011
0.007
0.003
0.003
species differences in genome size were tested using

GLM (general linear models) because of unbalanced data
design, and Tukey’s procedure was applied to compare
84178
77029
73446
71449
mean values. The Spearman-rank correlation coefficient

(CORR procedure) was used to test whether mean
genome size of the taxa was related to the geographical
location of the populations.
Škorničk. & M. Sabu, comb.
( ¼ C. caulina J. Graham)
(King ex Baker) Craib ex

Stahlianthus involucratus
( ¼ C. scaposa (Nimmo)
R. M. Sm. ( ¼ C. bhatii
(R.M.Sm.) Škorničk. &
R E S U LT S
RELATED GENERA
Kaempferia scaposa
Paracautleya bhatii
(J. Graham) Baker
Hitchenia caulina
Chromosome counts and ploidy levels

(Nimmo) Benth.
Chromosome numbers together with inferred ploidy levels

M. Sabu)
for the majority of the studied taxa (84 %) are given in

Table 2. In total, 50 plants (i.e. nearly one-third of all acces-
Loes.
nov.)
†
‡
§
sions) were analysed karyologically. Forty-one taxa


F I G . 1. Geographical origin of the plant samples analysed. (A) Species from genome group I (1Cx ¼ 0.30–0.33 pg) and x ¼ 7; (B) species from genome group
II (1Cx ¼ 0.36–0.39 pg) and x ¼ 7; (C) species from genome group III (1Cx ¼ 0.41–0.43 pg) and x ¼ 7; (D) species with 1Cx.0.83 pg and x ¼ 11 (circle,
Curcuma vamana; diamond, Stahlianthus involucratus). Symbol explanation (unless otherwise indicated): closed circles, hexaploids; open circles, nonaploids;
open diamonds, Curcuma-like species often placed into separate genera; open squares, high polyploids designated by a corresponding ploidy level.
(including undetermined samples) yielded definite chromo- of genome size estimates represent novel records; previous
some numbers, a preliminary count was obtained for one C-values were available for only ten taxa.
tentatively determined specimen (C. sp. ‘ranchi’ with Flow cytometric analyses yielded high-resolution histo-
2n.70), and a single chromosome record referring to grams (Fig. 3). Coefficients of variation (CVs) of G0/G1
Stahlianthus involucratus was taken from the literature (as peaks ranged from 0.89 to 5.93 % (mean 2.60) for
Kaempferia involucrata; Bisson et al., 1968). Six different Curcuma samples and from 1.13 to 6.54 % (mean 2.98)
chromosome numbers were identified (i.e. 2n ¼ 22, 42, 63, for the reference standard. An arbitrary threshold of 3.0 %
.70, 77 and 105). The majority of these are multiples of was not exceeded in 72 and 62 % of Curcuma and internal
x ¼ 7, which may be regarded as a genuine basic chromo-
some number. Consequently, plants with 42, 63, 77 and 105
somatic chromosomes correspond to hexaploids, nona-
ploids, 11-ploids and 15-ploids, respectively.
New chromosome numbers were found in two species,
C. oligantha (2n ¼ 77) and C. raktakanta (2n ¼ 105).
Both also represent new generic records and the latter is
the highest chromosome number so far determined in
Zingiberaceae. Micrographs documenting metaphase
chromosomes in C. vamana (2n ¼ 22) and C. raktakanta
(2n ¼ 105) are shown in Fig. 2.
F I G . 2. Chromosome complements of (A) C. vamana (species with the
smallest number of chromosomes) and (B) C. raktakanta (species with
Genome size variation the highest number of chromosomes) showing 22 and 105 somatic chromo-
somes, respectively. Scale bars ¼ 10 mm. In (A), two micrographs were
Table 2 summarizes the results for 161 samples belong- taken at different focal planes and computer-merged in order to achieve
ing to 51 taxa of Curcuma and related genera. The majority sufficient image sharpness.

F I G . 4. 2C-value distribution (DNA pg means) for 51 taxa investigated.
Symbol explanation: squares, diploids (2n ¼ 2x ¼ 22); diamonds, hexa-
ploids (2n ¼ 6x ¼ 42); triangles, nonaploids (2n ¼ 9x ¼ 63); circles,
higher polyploids designated by a corresponding ploidy level. Closed
symbols, Curcuma species; open symbols, closely related taxa often
placed into separate genera.
Nonetheless, there are some species alliances with phenoty-

pic similarities in which nuclear DNA amounts may pro-
vide a clue for accurate determination, C. cannanorensis
(2C ¼ 2.33 pg) – C. oligantha (2C ¼ 4.76 pg) being a repre-
sentative example (Fig. 5). In addition, genome size reflected
morphological variation in the C. aromatica complex, in
which three taxonomic entities with distinct DNA amounts
F I G . 3. (A) Representative flow cytometric histogram documenting
genome size determination in Curcuma oligantha (accession number
are currently recognized (Table 2). The genome size of a
73325) using Bellis perennis as internal reference standard. (B) Flow cyto- putative hybrid between C. angustifolia and C. montana
metric evidence for genome size variation in Curcuma longa [simul- was close to the mean of those of the parental species.
taneous analysis of accessions 71436 (2C ¼ 2.60 pg) and 86221-II Hexaploid Curcuma species showed marked variation in
(2C ¼ 2.85 pg)]. Plant nuclei were isolated, stained with propidium homoploid genome sizes, amounting to 45%. This variation
iodide and measured simultaneously.
was discontinuous and three groups of taxa with signifi-
cantly different Cx-values (P,0.0001, n ¼ 24) could be
standard runs, respectively. Between-day fluctuation in distinguished (Table 3). These groups corresponded well
FCM measurements due to instrument instability or non- to the geographical origin of the samples (Fig. 1). Higher
identical sample preparation was negligible, with standard polyploids mirrored this pattern and split into a genome
error of the mean ranging from 0.02 to 0.55 % of the esti- group I (1Cx ¼ 0.30– 0.32 pg) and a genome group III
mated 2C-value. Reliability of determined DNA amounts (1Cx.0.40 pg) cluster. To gain closer insights into the
was repeatedly confirmed in simultaneous FCM analyses, genome size variation, relationships between Cx-values
which gave two distinct peaks even in Curcuma samples and geographical locations of all samples with x ¼ 7 were
with only small differences in genome size (see Fig. 3B). examined. A significant negative correlation was found
Mean 2C-values varied from 1.66 pg in diploid C. vamana for both latitude (Spearman r ¼ – 0.50, P ¼ 0.0004, n ¼
(2n ¼ 2x ¼ 22) to 4.76 pg in C. oligantha (2n ¼ 11x ¼ 77), a 49) and longitude (r ¼ – 0.63, P,0.0001, n ¼ 49).
2.87-fold range (Fig. 4). Homoploid genome sizes ranged As diploids were found to possess a different basic
from 0.30 pg in several species to 1.56 pg in Stahlianthus chromosome number than polyploids (i.e. 11 vs. 7), poten-
involucratus, a 5.25-fold range. Most of the species with tial changes in homoploid genome size with respect to
more than one accession showed low intraspecific genome ploidy level were difficult to assess. Nevertheless, average
size variation (3.4 % on average). However, differences DNA content per chromosome in two diploid species
between maximum and minimum C-values that exceeded (C. vamana and Stahlianthus involucratus) exceeded the
4 % were observed in five species (C. prakasha, C. caesia, corresponding values in polyploid taxa.
C. raktakanta, C. montana and C. longa – arranged by With the exception of Stahlianthus involucratus, genome
increasing percentage variation) reaching 15.1 % in sizes of other species often placed in separate genera
C. longa (Fig. 3B). (Hitchenia caulina, Kaempferia scaposa, and Paracautleya
The value of FCM data for taxonomic purposes (species bhatii; Fig. 5) fitted well into the range of C-values of hex-
delineation) seems to be rather limited owing to the small aploid curcumas (they also shared the same number of
number of species-specific genome sizes (see Table 2). chromosomes).

F I G . 5. Phenotypic diversity of Curcuma species included in the study. (A) Curcuma vamana (accession number 84156); (B) C. cannanorensis (no.
84144); (C) C. raktakanta (no. 84120). (D) C. roscoeana (no. 73309); (E) C. oligantha (no. 73325); (F) C. ( ¼ Paracautleya) bhatii (no. 73446);
(G) C. ( ¼ Hitchenia) caulina (no. 84178); (H) C. kudagensis (no. 84152); (I) C. ( ¼ Kaempferia) scaposa (no. 77029). Individual plates not to scale.
DISCUSSION counts were each encountered in one taxon only (2n ¼

22 in C. vamana, 2n.70 in an undetermined sample
Comparison of chromosome numbers with
from eastern India, 2n ¼ 77 in C. oligantha and 2n ¼
previous investigations
105 in C. raktakanta; Figs 2 and 5). The last two
Our karyological analyses revealed six different chromo- records represent new chromosome numbers for the
some counts, 2n ¼ 22, 42, 63, .70, 77 and 105. The genus Curcuma, and 2n ¼ 105 is the highest somatic
somatic numbers 42 and 63 predominated and the other count recorded so far for the family Zingiberaceae.
TA B L E 3. Summary of chromosome counts and genome size were unsuccessful, either using direct karyological counting
estimates for species of Curcuma and related genera or using indirect FCM measurements. Finally, there are
determined in the present study apparent discrepancies in previous chromosome numbers
for C. malabarica (2n ¼ 42) and C. raktakanta (2n ¼ 63;
Ploidy level No. of No. of both published by Joseph et al., 1999). The counts here
or DNA chromosomes taxa/no. of 2C-value 1Cx-value
ploidy (2n) individuals range (pg) range (pg)
based on material from the type localities showed uniformly
2n ¼ 105. These taxa should be treated as one species
Curcuma (C. raktakanta) as also indicated by morphology.
Considering the discrepancies, it is likely that genuine

2x 22 1/1 1.66 0.83
6x, group I 42 10/42 1.79–1.96 0.30–0.33
C. malabarica and C. raktakanta were not involved in the
6x, group II 42 10/27 2.15–2.35 0.36–0.39 study of Joseph et al. (1999).
6x, group III 42 4/4 2.45–2.60 0.41–0.43 In addition to euploid chromosome numbers, several
9x 63 18/75 2.67–2.91 0.30–0.32 apparently aneuploid counts (i.e. 62, 64, 66, etc.) have
11x 77 1/1 4.76 0.43 been reported (see Table 1). Prana (1977) claimed that
cf. 12x .70 1/1 3.71 (0.31)
15x 105 1/5 4.57 0.30 this phenomenon may be associated with abnormal
mitosis occasionally encountered in Indonesian polyploid
Related genera
2x 22 1/1 3.11 1.56
curcumas. Daughter cells with unequal number of chromo-
6x 42 3/3 2.18–2.33 0.36–0.39 somes may be formed in this way and lead to incidence
of aneuploidy and/or chromosomal chimaeras, especially
in plants with predominantly vegetative reproduction.
However, no aneuploids were found in the present study.
No intraspecific variation in the chromosome number was Although we do not a priori reject such an option (e.g.
detected. Nevertheless, it should be noted that chromo- intraspecific variation in genome size observed in some
some counts were only analysed in more than one individ- Curcuma spp. may potentially refer to chromosomal hetero-
ual for eight taxa only. geneity), several records of aneuploidy are suspicious and
A targeted search for karyological data in Curcuma should be interpreted with caution until reliably confirmed.
showed that chromosome numbers have so far been pub- Karyological investigation of Curcuma spp. is rather chal-
lished for 40 identified species and four taxa of unclear lenging and errors may easily be introduced due to both
identity (see Table 1), 17 of which were also included in relatively high number and small size (mostly 0.5– 2 mm)
our study. Generally, there was agreement between the of chromosomes (Ramachandran, 1961; Apavatjrut, 1996;
data sets, although some discrepancies occurred. In particu- Joseph et al., 1999; Sirisawad et al., 2003). It should also
lar, several different chromosome counts for the same be noted that majority of doubtful counts were published
species have been reported in some cases, but this was several decades ago and have not been recorded since.
not confirmed in our study. Moreover, previous numbers
showed evidence of aneuploidy, a phenomenon not encoun-
Basic chromosome number and ploidy level variation
tered by us. Such incongruities may be explained either by
existence of karyological variation not sampled in the Since the pioneering karyological surveys, there has been
present study, species misidentification or a broader continuous dispute concerning the basic chromosome
species concept adopted in earlier studies; lack of herbar- number in Curcuma. Investigating 25 Zingiberaceae
ium vouchers, however, precludes verification of the taxo- species, including three curcumas, Raghavan and
nomic identity of previously studied plant material in Venkatasubban (1943) were the first to report x ¼ 21.
many cases. Sato (1948) suggested x ¼ 8 in his early work on
For example, two different numbers had previously been C. longa, but later claimed that two basic numbers (x ¼ 7
published for C. rubescens (2n ¼ 42 and 2n ¼ 63; Islam, and x ¼ 8) occur in the genus (Sato, 1960). Further
2004), but only the former was confirmed in our study. support for x ¼ 21 appeared in the study of
We believe that the plants with 63 chromosomes belonged Ramachandran (1961), who observed regular bivalent for-
to another species because C. rubescens was originally mation during meiosis in C. decipiens (2n ¼ 42) and a
described to produce both lateral and central inflorescences high frequency of trivalents in C. longa (under the name
by Roxburgh (1810), a feature common in hexaploids but C. domestica) with 2n ¼ 63. In line with Venkatasubban
unknown in any nonaploid species. A parallel situation (1946), Ramachandran considered x ¼ 21 a secondary
appears to exist in C. mangga. Again, we found only 42 number possibly derived from a combination of x ¼ 9 and
chromosomes in this species, but found 2n ¼ 63 in an unde- x ¼ 12, which are common in several genera of
termined, though closely related, specimen (accession no. Zingiberaceae. Yet another basic chromosome number,
86306). We have some doubts about the degree of chromo- x ¼ 16, has been proposed and adopted by some researchers
somal variation in C. aromatica ( published 2n ¼ 42, 63, 84 (e.g. Sharma and Bhattacharya, 1959). However, x ¼ 21
and 86). Current taxonomic investigations revealed that this became the most commonly accepted basic chromosome
is a complex group composed of several species that possess number in Curcuma (Ramachandran, 1961; Prana, 1977;
either 42 or 63 chromosomes according to the present results. Prana et al., 1978; Ardiyani, 2002; Islam, 2004).
Despite representative sampling, attempts to find other It should be noted that the lower alternative, x ¼ 7, is not
chromosome numbers within the C. aromatica aggregate in conflict with the large majority of published somatic
counts. Actually, this value fits better with the range of histories, as also supported by their distinct distribution pat-
chromosome numbers currently known (see our prime terns (see below).
count 2n ¼ 77). We therefore believe that x ¼ 7 should be Intraspecific genome size variation was low in most taxa
considered a primary basic chromosome number, at least in which multiple individuals were analysed. However,
for the majority of Indian Curcuma species (belonging to values above 4 % (i.e. beyond potential instrumental fluctu-
the subgenus Curcuma). Grant (1982) regarded x ¼ 7 and ation) were encountered in five species. When C. longa with
10 as the most common basic chromosome numbers in about 1.15-fold divergence in C-value is excluded, the
monocots so the present findings are in concordance with extent of intraspecific variation was comparable in all cyto-
a large body of evidence. Following this calculation, types: hexaploids, 0.0 – 6.1 %; nonaploids, 0.5 – 4.9 %; and a

species investigated here correspond to 6x, 9x, 11x and 15-ploid, 5.1%. Intraspecific variation in nuclear DNA
15x cytotypes. In addition, dodecaploid (2n ¼ 84) plants content has been controversial since the early studies on
from subgenus Curcuma were reported by earlier research- genome size. This debate has been fuelled by numerous
ers (Table 1). early reports of intraspecific variation, many of which
It appears that there are some Curcuma species in which were dismissed in subsequent investigations using the best
somatic chromosome numbers (2n ¼ 20, 22, 24, 32, 34, 36 practice methodology (Greilhuber, 2005). Several sources
and 38) do not fit into the series based on x ¼ 7. However, of artefactual variation have been identified: (1) instrumen-
they all belong to the subgenus Hitcheniopsis, which tal or methodological errors; (2) disturbing effects of sec-
encompasses mainly Thai species such as C. alismatifolia, ondary metabolites with potential seasonal fluctuation
C. gracillima, C. harmandii, C. parviflora, C. rhabdota (e.g. Walker et al., 2006); (3) differences in measurements
and C. thorelii (see Table 1), and which differs markedly among different laboratories (Doležel et al., 1998); and (4)
in morphological traits from the nominate subgenus. From taxonomic heterogeneity of the material under investigation
the present species set, only C. vamana (2n ¼ 22) belongs (Murray, 2005). As a result, the concept of genome size
to subgenus Hitcheniopsis. We are convinced that this stability has gained broader support. Examples have none-
species is only distantly related to the other investigated theless been accumulated in recent years of some genome
taxa (subgenus Curcuma), and its basic chromosome size variation in FCM assays despite meticulous method-
number is most plausibly x ¼ 11, as also documented for ology (Obermayer and Greilhuber, 2005; Šmarda and
several other genera of Zingiberaceae (e.g. Kaempferia, Bureš, 2006). Several Curcuma species are known to
Zingiber and Renealmia). contain, particularly in rhizomes, phenolic secondary
It is plausible that both subgenera Curcuma and metabolites such as curcumins (Lubis, 1968; Prana,
Hitcheniopsis represent independent evolutionary units 1977). Although phenolics may bias genome size estimates
with distinct basic chromosome numbers. In addition, x ¼ (Greilhuber, 1998; Walker et al., 2006), we believe that our
7 may be considered a subgenus-specific cytological FCM measurements are accurate for several reasons. Low
marker for the former. coefficients of variation were achieved, which are not com-
patible with the presence of interfering metabolites. Nuclear
suspensions were clear, lacking any coatings of debris that
might indicate a negative effect of metabolites (Loureiro
Inter- and intraspecific variation in genome size
et al., 2006). Repeated measurements of the same sample
The taxa involved in the present study differed 2.87-fold yielded nearly identical genome size values (average differ-
in their holoploid genome sizes (2C ¼ 1.66– 4.76 pg). This ence 0.8 %) even if particular FCM analyses were performed
range is slightly below the mean for other plant genera in different years. Between-plant differences remained stable
(3.28-fold based on data from the Plant DNA C-values data- also in studies using DAPI (an AT-selective fluorochrome
base; Bennett and Leitch, 2005a). There was no clear gap in less sensitive to secondary metabolites than the intercalating
nuclear DNA C-values between hexaploid and nonaploid propidium iodide). Also, a narrow species concept was
plants and a major discontinuity in these cytotypes actually adopted to guarantee taxonomic homogeneity of the plant
occurred between two groups of hexaploids (Fig. 4). These material. Moreover, co-processed samples with different
results indicate that DNA content alone is often insufficient genome sizes always gave two distinct peaks, which is the
to distinguish between 6x and 9x cytotypes, and FCM most convincing evidence for genuine differences in DNA
measurements should always be accompanied by chromo- content (Greilhuber, 2005).
some counts. The C-values of diploid and high polyploid The reasons for intraspecific genome size variation in
taxa were more distinct, but again FCM measurements Curcuma remain unknown. Aneuploidy or presence of
alone may not be an accurate indicator of ploidy level, B-chromosomes may lead to heterogeneity in nuclear
as illustrated by 11-ploid C. oligantha and 15-ploid DNA amount, but this explanation seems rather unlikely
C. raktakanta with reverse total amounts of nuclear DNA as only euploid numbers were revealed in our study and,
(Fig. 4). more importantly, two accessions of C. longa with more
By contrast, homoploid genome sizes formed three well- than 9 % genome size variation possessed the same
defined clusters with significantly different values, namely number of chromosomes (see Table 2). Plausibly, intraspe-
a genome group I (1Cx ¼ 0.30– 0.33 pg; 29 taxa), a cific variation may be related to a long-term cultivation and
genome group II (1Cx ¼ 0.36– 0.39; 13 taxa) and a targeted selection of desirable genotypes in several
genome group III (1Cx ¼ 0.41– 0.43 pg; five taxa). We Curcuma species, C. longa in particular. India is respon-
assume that these groups may have different evolutionary sible for around 90 % of turmeric production worldwide,
and this species has been widely used in India since Vedic from the methodological point of view. Unfortunately,
times. Perhaps the variation may have adaptive value, as genome size estimates were not linked to a particular her-
previously documented in another crop, Zea mays barium voucher, which lessens their value. We had the
(Rayburn and Auger, 1990). Murray (2005) argued that opportunity to examine several well-prepared herbarium
intraspecific genome size variation may indicate incipient specimens prepared from Islam’s living material and
speciation; the blurred species boundaries in several encountered some misidentifications (e.g. for C. aeruginosa
Curcuma alliances may support such a hypothesis. A and C. zedoaria).
third explanation takes into account heterochromatic poly- In comparison with Islam’s work (Islam, 2004), our esti-
morphism. In vegetatively propagating lines, strains with mates for ten species in common were on average 14 %

smaller and larger telomeric or centromeric heterochroma- lower, whereas we determined genome size to be about
tin regions may occur (J. Greilhuber, University of 11 % higher in C. zantorrhiza than that reported by
Vienna, Austria, pers. comm.) leading to greater genome Bharathan et al. (1994). Plausibly, different reference stan-
sizes. dards could be responsible for such divergences (i.e.
Raphanus sativus – Islam, 2004; chicken red blood cells
– Bharathan, 1994).
Comparison of genome sizes in Curcuma with those
in other members of Zingiberaceae and other monocots
Species distribution and origin of high polyploids
More than three-quarters of the taxa (39 out of 51) in the
with respect to homoploid genome size
present study possessed very small genomes defined as
1C1.40 pg (Leitch et al., 1998), whereas the remaining Excluding the distantly related C. vamana, species
12 taxa had small genomes (i.e. 1C ¼ 1.41– 3.50 pg). Low with the lowest number of chromosomes (2n ¼ 6x ¼ 42)
nuclear DNA content in Curcuma spp. is in line with previous clustered in three groups with significantly different
records for other members of Zingiberaceae and related Cx-values (Table 3), which also showed a distinct geo-
families. The Plant DNA C-values database (Bennett and graphical pattern (Fig. 1). Group I (1Cx ¼ 0.30 – 0.33
Leitch, 2005a) includes measurements for three members pg) contained ten taxa, the majority of which occurred
of Zingiberaceae with 1C-values varying from 1.30 pg in in north-east India, group II (1Cx ¼ 0.36 – 0.39 pg) con-
Curcuma zantorrhiza (misspelled as C. xanthorrhiza) to tained ten taxa with a centre of distribution in the
6.03 pg in Zingiber officinale. Very small genomes Western Ghats (west and south-west India) and Central
were also revealed in all but one (Lowiaceae with 1C ¼ India and group III (1Cx ¼ 0.41 – 0.43 pg) contained
3.55 pg) families from the order Zingiberales, including four taxa occurring either in south-west or north-east
Marantaceae (14 species, 1C ¼ 0.33– 0.68 pg), Musaceae India. A link between genome size and geographical
(six species, 1C ¼ 0.58 – 0.61 pg), Heliconiaceae (1C ¼ location was also confirmed by correlation analysis,
0.45 pg), Strelitziaceae (1C ¼ 0.58 pg), Cannaceae (1C ¼ which revealed highly significant negative relationships
0.72 pg) and Costaceae (1C ¼ 1.00 pg). between Cx-values and both latitude and longitude of
Although only one Curcuma record is included in the the localities.
Plant DNA C-values database, more genome size esti- Homoploid genome sizes of all but one (C. oligantha)
mates have been published (Table 1). Das et al. (1999) high-polyploid taxa matched perfectly the Cx-values of
used Feulgen densitometry to analyse three Curcuma hexaploids from group I. This allows us to hypothesize
spp.: C. caesia (2n ¼ 22, 4C ¼ 3.120 pg), C. amada that these hexaploids have played a key role in the evol-
(2n ¼ 40, 4C ¼ 4.234 pg) and C. longa (2n ¼ 48, 4C ¼ ution of polyploidy in Indian Curcuma spp. Nonaploid
5.100 – 5.263 pg). Curcuma longa was also investigated cytotypes probably originated by a fusion of reduced
by Nayak et al. (2006) who reported 4C-values from and unreduced gametes of hexaploids, either within or
4.30 to 8.84 pg (i.e. 2.06-fold variation) in 17 cultivars. between species, giving rise to auto- or allopolyploids.
However, both these studies come from the same labora- Within Zingiberaceae, regular production of gametes
tory and we believe the results should be treated with with the somatic number of chromosomes has been
caution. First, chromosome counts have not been con- reported in the genus Globba (Takano and Okada,
firmed by other researchers. Moreover, the authors used 2002), which shows similar ploidy composition to
hot hydrolysis, which is strongly discouraged owing to Curcuma. Both nonaploid (unreduced gamete) and hexa-
the very short optimum treatment that is difficult to ploid (reduced and unreduced gamete) plants were prob-
control (Greilhuber, 2005). No fixative solution was speci- ably involved in the genesis of rare dodecaploid and
fied, and if a standard acetic acid/alcohol fixation was 15-ploid taxa. The vast majority of nonaploid species
applied to plant samples rich in phenolics, stoichiometry undergo asexual propagation via rhizome branching and
of the Feulgen staining was likely to be distorted (the produce a high percentage of aborted pollen grains
so-called ‘self-tanning’ effect; Greilhuber, 1986, 1988). (Prana, 1977; Nasir Uddin, 2000). It is plausible,
Feulgen methodology is also not recommended for conven- however, that even a small fraction of viable pollen may
tional chromosome counting in Curcuma as it does not eventually lead to the formation of higher polyploids.
allow good spreading of the root tips issue on slides The origin of 11-ploid C. oligantha cannot be explained
(Islam, 2004) and gives rather pale staining. By contrast, with certainty given the current stage of knowledge.
the FCM data published for Curcuma spp. collected in Homoploid genome size (1Cx ¼ 0.43 pg) of this species
Bangladesh by Islam (2004) seem to be accurate, at least fits in the range of a genome group III.
Genome size and chromosome numbers as supportive Hitchenia caulina, Kaempferia scaposa and Paracautleya
markers for delimitation of Curcuma species bhatii, reveal a similar situation.
One aim of the present study was to assess whether
Chromosome numbers and/or ploidy levels have long
ploidy and genome size data provide additional information
been utilized as an efficient taxonomic marker, helping
useful for taxonomic decision-making. Genome sizes of the
to delimit boundaries between various taxonomic cat-
three controversial species (Hitchenia caulina: 2C ¼ 2.25
egories or reveal cryptic taxa (Stace, 2000). In addition,
pg, Kaempferia scaposa: 2C ¼ 2.33 pg and Paracautleya
the last decade has seen significant progress in the appli-
bhatii: 2C ¼ 2.18 pg) were found to match hexaploid
cation of FCM to exploit differences in nuclear DNA
Curcuma taxa from the genome group II. In addition, they

content, which can in some cases be used as a supportive
all had the hexaploid number of chromosomes (2n ¼ 42)
marker for distinguishing between closely related taxa at
and similar geographical distribution in the Western
both heteroploid and homoploid levels (e.g. Mahelka
Ghats, a biodiversity hotspot for Curcuma. These lines of
et al., 2005).
evidence together with a lack of clear-cut morphological
As Curcuma is a taxonomically challenging group, the
traits support inclusion of the taxa in question in a
usefulness of genome size and chromosome counts as sup-
broadly defined Curcuma (see Škorničková and Sabu,
portive markers for taxon determination was assessed.
2005a, for the latest circumscription of the genus).
Although the low C-value variation (2.87-fold) and the
Although valid name combinations in Curcuma already
small number of species-specific DNA amounts reduce
exist for both Hitchenia caulina (originally described
their utility, some alliances with morphological similarities
as C. caulina) and Paracautleya bhatii (C. bhatii, in
have been identified in which cytogenetic data may nonethe-
Škorničková and Sabu, 2005a), Kaempferia scaposa has
less support taxonomic decisions. For example, C. oligantha
never been included in Curcuma, perhaps due to the
(described from Sri Lanka) and C. cannanorensis (described
absence of pouches formed by the bracts and its pure
from Kerala, western coast of South India) have traditionally
white flowers with extremely long floral tubes (Fig. 5).
be merged as one taxon due to their overall phenotypic
However, the presence of pouches is neither a unique nor
resemblance (Bhat, 1987; Mangaly and Sabu, 1993; Sabu,
a universal feature for the genus Curcuma (Kress et al.,
2006). However, the present examination of living plants
2002; Škorničková and Sabu, 2005a) and the distinct
in natural conditions revealed constant, though minor, mor-
flower morphology and colour may represent an adaptation
phological divergences (see Fig. 5), along with differences
to night pollination, as also observed in another ginger
in ecological requirements. Distinctiveness of both species
species with nocturnal anthesis, Leptosolena haenkei
was further corroborated by distinct chromosome numbers
(Funakoshi et al., 2005). Transfer of Kaempferia scaposa
(2n ¼ 77 in the former, 2n ¼ 42 in the latter). Similarly, it
to the genus Curcuma requires a new name combination,
was found that C. aromatica is a species complex comprising
which is proposed below.
three distinct morphotypes with non-overlapping genome
By contrast, Stahlianthus involucratus possesses unique
sizes, including hexaploids (2C ¼ 1.86 pg) from south
holoploid and homoploid genome sizes (1C ¼ 1Cx ¼ 1.56
India, nonaploids (2C ¼ 2.83 pg) from north-east India and
pg), dissimilar to any other species involved in the
nonaploids (2C ¼ 2.68 pg) from eastern India. Taxon
current study. The diploid number of chromosomes (2n ¼
descriptions and clarification of the nomenclatural issues
22) was shared only with south Indian C. vamana, but
are in progress. Curcuma reclinata (2C ¼ 2.29 pg),
this species has a different Cx-value and overall mor-
C. decipiens (2C ¼ 2.35 pg) and C. inodora (2C ¼ 2.29
phology. We therefore keep this taxon in a separate genus
pg) are difficult to define morphologically and have more
at this point, but targeted investigation of Thai and south-
or less overlapping distributions. Cytogenetic data do not
east Asian members of the genera Curcuma and
provide reliable data for species-level determination and we
Kaempferia is necessary to arrive at a final decision con-
believe that they should be merged as one taxon (i.e.
cerning the validity of this genus.
C. reclinata Roxb.).
Curcuma scaposa (Nimmo) Škorničk. & M.Sabu,
comb. nov.
Taxonomic position of Curcuma-like species placed Basionym Hedychium scaposum Nimmo, in Graham,
into separate genera Cat. Pl. Bombay 205 (1839);;Monolophus scaposus
(Nimmo) Dalzell, Hooker’s J. Bot. Kew Gard. Misc. 2:
The taxonomic position of some species with a 143 (1850);;Kaempferia scaposa (Nimmo) Benth. Gen.
Curcuma-like morphology has long been discussed. Pl. 3: 642 (1883).
Traditionally, they have been placed into separate genera Type: [India, Maharashtra], Western Ghats, Lonavlie,
such as Hitchenia, Kaempferia, Paracautleya and September 1878, G. King s.n. (Neotype: BM!, designated
Stahlianthus. However, a phylogenetic analysis of the here. Isoneotype: K!).
tribe Zingibereae based on internally transcribed spacers
and trnL – trnF gene sequences (Ngamriabsakul et al.,
2004) did not support their independent generic status as
ACK N OW L E D G E M E N T S
these taxa were nested within the Curcuma complex. Our
molecular analyses (T. Fér et al., Charles University, A long-term stay of J.L.-Š. at the Calicut University,
Prague, Czech Republic, unpubl. res.) based on trnL –trnF Kerala, India, was supported by fellowships from the
gene sequences of a number of Indian taxa, including Indian Council of Cultural Relationships and the Ministry
of Education, Youth and Sport of the Czech Republic. We of the 2nd Symposium on the Family Zingiberaceae. Guangzhou:
thank the authorities of the Ministry of Environment and Zhongshan University Press Press, 107–111.
Eksomtramage L, Sirirugsa P, Jivanit P, Maknoi C. 2002. Chromosome
Forests, Government of India, and the Botanical Survey counts of some zingiberaceous species from Thailand.
of India for granting collecting permits and permits to Songklanakarin Journal of Science and Technology 24: 311– 319.
access Indian herbaria. We are also grateful to Johann Funakoshi H, Kress JW, Škorničková J, Liu A, Inoue K. 2005. Return
Greilhuber (Vienna) for critical comments on intraspecific from the lost: rediscovery of the presumed extinct Leptosolena
(Zingiberaceae) in the Philippines and its phylogenetic placement in
genome size variation, and Ilia Leitch (Kew) for providing gingers. Acta Phtyotaxonomica et Geobotanica 56: 41–53.
some literature. We thank the handling editor Michael Fay Grant V. 1982. Periodicities in the chromosome numbers of the angios-
(Kew) and two anonymous reviewers for useful remarks on perms. Botanical Gazette 143: 379–389.

the manuscript. This project was supported by the Grant Greilhuber J. 1986. Severely distorted Feulgen DNA amounts in Pinus
Agency of the Academy of Sciences of the Czech Republic (Coniferophytina) after nonadditive fixations as a result of meriste-
matic self-tanning with vacuole contents. Canadian Journal of
( project no. B6407401), the Department of Science and Genetics and Cytology 28: 409– 415.
Technology, Government of India (Order No. SP/SO/ Greilhuber J. 1988. ‘Self-tanning’ – a new and important source of stoi-
A-20/99 dt. 9.11.2001), the European Commission’s chiometric error in cytophotometric determination of nuclear DNA
Research Infrastructure Action via the SYNTHESYS content in plants. Plant Systematics and Evolution 158: 87–96.
Project (GB-TAF-606), the Ministry of Education, Youth Greilhuber J. 1998. Intraspecific variation in genome size: a critical reas-
sessment. Annals of Botany 82 (Suppl. A): 27– 35.
and Sport of the Czech Republic (MSM 0021620828) and Greilhuber J. 2005. Intraspecific variation in genome size in angiosperms:
the Academy of Sciences of the Czech Republic identifying its existence. Annals of Botany 95: 91–98.
(AV0Z60050516). Islam MA. 2004. Genetic diversity of the genus Curcuma in Bangladesh
and further biotechnological approaches for in vitro regeneration
and long-term conservation of C. longa germplasm. PhD thesis,
University of Hannover, Germany.
Joseph R, Joseph T, Joseph J. 1999. Karyomorphological studies in the
L IT E RAT URE C IT E D genus Curcuma Linn. Cytologia 64: 313–317.
Apavatjrut P, Sirisawad T, Sirirugsa P, Voraurai P, Suwanthada C. Kress WJ, Prince LM, Williams KJ. 2002. The phylogeny and a new
1996. Studies on chromosome number of seventeen Thai Curcuma classification of the gingers (Zingiberaceae): evidence from molecular
species. Proceedings of 2nd National Conference on Flower and data. American Journal of Botany 89: 1682– 1696.
Ornamental Plant 2: 86– 99. Larsen K, Lock JM, Maas H, Maas PJM. 1998. Zingiberaceae. In:
Ardiyani M. 2002. Systematic study of Curcuma L.: turmeric and its allies. Kubitzki K., ed. The families and genera of vascular plants, vol. 4.
PhD thesis, University of Edinburgh, Scotland. Berlin, Springer-Verlag, 474–495.
Beltran IC, Kam YK. 1984. Cytotaxonomic studies in the Zingiberaceae. Leitch IJ, Chase MW, Bennett MD. 1998. Phylogenetic analysis of DNA
Notes from the Royal Botanic Garden Edinburgh 41: 541–559. C-values provides evidence for a small ancestral genome size in flow-
Bennett MD, Leitch IJ. 2005a. Plant DNA C-values Database (Release ering plants. Annals of Botany 82 (Suppl. A): 85– 94.
4.0, Oct. 2005), http://www.kew.org/genomesize/homepage (accessed Lim SN. 1972a. Cytogenetics and taxonomy of the genus Globba L.
5 January, 2007). (Zingiberaceae) in Malaya. II. Cytogenetics. Notes Royal Botanic
Bennett MD, Leitch IJ. 2005b. Nuclear DNA amounts in angiosperms: Gardens Edinburgh 31: 271– 284.
progress, problems, and prospects. Annals of Botany 95: 45–90. Lim SN. 1972b. Cytogenetics and taxonomy of the genus Globba L.
Bharathan G, Lambert G, Galbraith DW. 1994. Nuclear DNA content (Zingiberaceae) in Malaya. IV. Distribution in relation to polyploidy.
of monocotyledons and related taxa. American Journal of Botany Gardens’ Bulletin Singapore 26: 115– 126.
81: 381 –386. Loureiro J, Rodriguez E, Doležel J, Santos C. 2006. Flow cytometric
Bhat KG. 1987. Curcuma oligantha Trimen (Zingiberaceae) – a new and microscopic analysis of the effect of tannic acid on plant nuclei
record for India. Indian Journal of Forestry 10: 66– 68. and estimation of DNA content. Annals of Botany 98: 515–527.
Bisson S, Guillemet S, Hamel J-L. 1968. Contribution a l’étude caryo- Lubis I. 1968. Phenolic compounds of Curcuma. Annales Bogoriensis 4:
taxonomique des Scitaminées. Memoires du Museum National 219– 225.
d’Histoire Naturelle. Série B. Botanique 18: 59–145. Mahelka V, Suda J, Jarolı́mová V, Trávnı́ček P, Krahulec F. 2005.
Chakravorti AK. 1948. Multiplication of chromosome numbers in Genome size discriminates between closely related taxa Elytrigia
relation to speciation in Zingiberaceae. Science and Culture 14: repens and E. intermedia (Poaceae: Triticeae) and their hybrid.
137– 140. Folia Geobotanica 40: 367– 384.
Chen ZY, Chen SJ. 1984. A report on chromosome numbers of Chinese Mangaly JK, Sabu M. 1993. A taxonomic revision of the South Indian
Zingiberaceae. Guihaia 4: 13– 18. species of Curcuma Linn. (Zingiberaceae). Rheedea 3: 139–171.
Chen ZY, Chen SJ, Huang XX, Huang SF. 1988. A report on chromo- Mukherjee I. 1970. Chromosome studies of some species of Hedychium.
some numbers on Chinese Zingiberaceae (5). Guihaia 8: 143– 147. Botanical Magazine Tokyo 83: 237–241.
Das AB, Rai S, Das P. 1999. Karyotype analysis and cytophotometric esti- Murray BG. 2005. When does intraspecific C-value variation become tax-
mation of nuclear DNA content in some members of the onomically significant? Annals of Botany 95: 119– 125.
Zingiberaceae. Cytobios 97: 23– 33. Nambiar MC, Pillai PKT, Sharma YN. 1982. Seedling propagation in
Doležel J, Doleželová M, Novák FJ. 1994. Flow cytometric estimation of turmeric (Curcuma aromatica Salisb.). Journal of Plantation Crops
nuclear DNA amount in diploid bananas (Musa acuminata and 10: 81– 85.
M. balbisiana). Biologia Plantarum 36: 351–357. Nasir Uddin S. 2000. The origin of turmeric: the domestication of an
Doležel J, Greilhuber J, Lucretti S, Meister A, Lysák MA, Nardi L, important ginger spice. MSc thesis, University of Edinburgh,
Obermayer R. 1998. Plant genome size estimation by flow cytometry: Scotland.
inter-laboratory comparison. Annals of Botany 82 (Suppl. A): 17–26. Nayak S, Naik PK, Acharya LK, Pattnaik AK. 2006. Detection and
Eksomtramage L, Boontum K. 1995. Chromosome counts of evaluation of genetic variation in 17 promising cultivars of turmeric
Zingiberaceae. Songklanakarin Journal of Science and Technology (Curcuma longa L.) using 4C nuclear DNA content and RAPD
17: 291 –297. markers. Cytologia 71: 49–55.
Eksomtramage L., Sirirugsa P, Mayakul S. 1996a. Chromosome Ngamriabsakul C, Newman MF, Cronk QCB. 2004. The phylogeny of
numbers of some Thai Zingiberaceae. Songklanakarin Journal of tribe Zingibereae (Zingiberaceae) based on ITS (nrDNA) and trnL-F
Science and Technology 18: 153–159. (cpDNA) sequences. Edinburgh Journal of Botany 57: 39–61.
Eksomtramage L, Sirirugsa P, Mayakul S. 1996b. Chromosome counts of Obermayer R, Greilhuber J. 2005. Does genome size in Dasypyrum vil-
Thai Zingiberaceae. In: Wu TL, Wu QG, Chen ZY, eds. Proceedings losum vary with fruit colour? Heredity 95: 91–95.
Otto F. 1990. DAPI staining of fixed cells for high-resolution flow cytome- Sirirugsa P. 1996. The genus Curcuma of Thailand. In: Wu TL, Wu QG,
try of nuclear DNA. In: Crissman HA, Darzynkiewicz Z, eds. Chen ZY, eds. Proceedings of the 2nd Symposium on the family
Methods in cell biology, vol. 33. New York: Academic Press, Zingiberaceae. Guangzhou: Zhongshan University Press, 39– 46.
105–110. Sirisawad T, Sirirugsa P, Suwanthada C, Apavtjrut P. 2003.
Paisooksantivatana Y, Thepsen O. 2001. Phenetic relationship of some Investigation of chromosome numbers in 20 taxa of Curcuma. In:
Thai Curcuma species (Zingiberaceae) based on morphological, paly- Chantaranothai P, Larsen K, Sirirugsa P, Simpson D, eds.
nological and cytological evidence. Thai Journal of Agricultural Proceedings of the 3rd Symposium on the family Zingiberaceae.
Science 34: 47– 57. Khon Kaen: Applied Taxonomic Research Centre, Khon Kaen
Poulsen AD. 1993. Two new species of Boesenbergia (Zingiberaceae) University, 54–62.
from Borneo. Nordic Journal of Botany 13: 289 –294. Škorničková J, Sabu M. 2005a. The recircumscription of Curcuma L. to
Prana MS. 1977. Studies on some Indonesian Curcuma species. PhD include the genus Paracautleya R.M.Sm. Gardens’ Bulletin

thesis, University of Birmingham, England. Singapore 57: 37–46.
Prana MS, Sastrapradja S, Hawkes JG, Lubis I. 1978. A cytological Škorničková J, Sabu M. 2005b. Curcuma roscoeana Wall.
study of some Indonesian Curcuma species. Journal of Root Crops (Zingiberaceae) in India. Gardens’ Bulletin Singapore 57: 187–198.
4: 31–35. Škorničková J, Sabu M. 2005c. The identity and distribution of Curcuma
Raghavan RS, Arora CM. 1958. Chromosome numbers in Indian medic- zanthorrhiza Roxb. (Zingiberaceae) Gardens’ Bulletin Singapore 57:
inal plants. II. Proceedings of Indian Academy of Sciences Series B 199–210.
47: 352– 358. Škorničková J, Sabu M, Prasanthkumar MG. 2003a. A new species of
Raghavan TS, Venkatasubban KR. 1943. Cytological studies in the Curcuma L. (Zingiberaceae) from Mizoram. Gardens’ Bulletin
family Zingiberaceae with special reference to chromosome number Singapore 55: 89–95.
and cyto-taxonomy. Proceedings of Indian Academy of Sciences Škorničková J, Sabu M, Prasanthkumar MG. 2003b. Curcuma codo-
Series B 17: 118– 132. nantha (Zingiberaceae) – a new species from the Andaman Islands,
Ramachandran K. 1961. Chromosome numbers in the genus Curcuma India. Gardens’ Bulletin Singapore 55: 219– 228.
Linn. Current Science 30: 194– 196. Škorničková J, Sabu M, Prasanthkumar MG. 2004. Curcuma mutabilis
Ramachandran K. 1969. Chromosome numbers in Zingiberaceae. (Zingiberaceae): a new species from South India. Gardens’ Bulletin
Cytologia 34: 213– 221.
Singapore 56: 43–54.
Rayburn AL, Auger JA. 1990. Nuclear DNA-content variation in the
Škorničková J, Rehse T, Sabu M. 2007. Other economically important
ancient indigenous races of Mexican maize. Acta Botanica
Curcuma species. In: Ravindran PN, Babu KN, Sivaraman K., eds.
Neerlandica 39: 197–202.
Turmeric: the genus Curcuma. Boca Raton, FL: CRC Press, 451– 467.
Roxburgh W. 1810. Descriptions of several of the monandrous plants of
Šmarda P, Bureš P. 2006. Intraspecific DNA content variability in
India. Asiatick Researches 11: 318– 362.
Festuca pallens on different geographical scales and ploidy levels.
Sabu M. 2006. Zingiberaceae and Costaceae of South India. Feroke:
Indian Association of Angiosperm Taxonomy. Annals of Botany 98: 665– 678.
Saensouk S, Chantaranothai P. 2003. The family Zingiberaceae in Phu Smith RM. 1981. Synoptic keys to the genera of Zingiberaceae pro parte.
Phan National Park. In: Chantaranothai P, Larsen K, Sirirugsa P, Edinburgh: Royal Botanic Garden Edinburgh.
Simpson D, eds. Proceedings of the 3rd Symposium on the family Stace CA. 2000. Cytology and cytogenetics as a fundamental taxonomic
Zingiberaceae. Khon Kaen: Applied Taxonomic Research Centre, resource for the 20th and 21st centuries. Taxon 49: 451– 477.
Khon Kaen University, 16–25. Suguira T. 1931. A list of chromosome numbers in angiospermous plants.
Saensouk S, Sumonthip B, Luangpirom A. 1998. Chromosome numbers Botanical Magazine Tokyo 45: 353–355.
of some Zingiberaceae in Phu Phan National Park. In: 24th Congress Suguira T. 1936. Studies on the chromosome numbers in higher plants,
on Science and Technology of Thailand, 19–21 October 1998. . Cited with special reference to cytokenesis, I. Cytologia 7: 544– 595.
in: Saensouk S, Chantaranothai P. 2003. Takano A. 2001. Cytological analyses of 19 taxa in Globba
Sarkar AK. 1990. Cytological investigation of certain members of (Zingiberaceae). Acta Phytotaxonomica et Geobotanica 52: 65– 74.
Zingiberaceae Lindl. to ascertain their taxonomic affinities. Takano A, Okada H. 2002. Multiple occurrence of triploid formation in
Proceedings of Indian Science Congress 77: 150. Globba (Zingiberaceae) from molecular evidence. Plant Systematics
Sato D. 1948. Karyotype and systematics of Zingiberales. Japanese and Evolution 230: 143– 159.
Journal of Genetics 23: 44–45. Venkatasubban KR. 1946. A preliminary survey of chromosome numbers
Sato D. 1960. The karyotype analysis in Zingiberales with special refer- in Scitamineae of Bentham & Hooker. Proceedings of Indian
ence to the protokaryotype and stable karyotype. Scientific Papers Academy of Sciences Series B 23: 281–300.
of the College of General Education, University of Tokyo 10: Walker DJ, Monino I, Correal E. 2006. Genome size in Bituminaria
225–243. bituminosa (L.) C. H. Stirton (Fabaceae) populations: separation of
Schumann K. 1904. Zingiberaceae. In: Engler HGA, ed. Das “true” differences from environmental effects on DNA determination.
Pflanzenreich 46: 1 –458. Environmental and Experimental Botany 55: 258– 265.
Sharma AK, Bhattacharya NK. 1959. Cytology of several members of Weerapakdee W, Krasaechai A. 1997. Collection and development
Zingiberaceae. La Cellule 59: 297–346. studies of certain Curcuma spp. Journal of Agriculture 13: 127– 136.

Chromosome Numbers and Genome Size Variation in Indian Species of Curcuma (Zingiberaceae)

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Chromosome Numbers and Genome Size Variation in Indian Species of Curcuma (Zingiberaceae)

Uploaded by

Copyright:

Available Formats

Annals of Botany 100: 505–526, 2007

doi:10.1093/aob/mcm144, available online at www.aob.oxfordjournals.org

Chromosome Numbers and Genome Size Variation in Indian Species of Curcuma

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

IN TROD UCT IO N Several taxonomic and biological problems have hin-

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

subgen. Curcuma K.Schum.

Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

C. heyneana Valeton & Zijp 63 Indonesia Prana (1977, 1978)

Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma

* Genome sizes expressed in 2C-values unless indicated otherwise.

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

71439B 2.782+ 1.39 1360 9 0.31 L Meghalaya,

Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Leong-Škorničková et al. — Chromosome Numbers and Genome Size in Curcuma

and nuclei were gently resuspended in 100 mL of fresh Otto I

at room temperature, after which fluorescence intensity of

(Partec GmbH, Münster, Germany) equipped with a 532-nm

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Each plant was re-analysed at least three times on different

min. value) exceeded 2 %, the outlying value was discarded

L. ‘Stupické polnı́ tyčkové rané’ (2C ¼ 2.11 pg) and Bellis

sizes, respectively, in order to minimize standard-to-sample

replications on different days. The total number of flow cyto-

root tips of the cultivated plants. Samples were pretreated

temperature), fixed in a 3:1 mixture of ethanol and acetic

was determined in 5 – 10 complete well-spread mitotic plates

species differences in genome size were tested using

mean values. The Spearman-rank correlation coefficient

(King ex Baker) Craib ex

Chromosome counts and ploidy levels

Chromosome numbers together with inferred ploidy levels

for the majority of the studied taxa (84 %) are given in

sions) were analysed karyologically. Forty-one taxa

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Nonetheless, there are some species alliances with phenoty-

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

DISCUSSION counts were each encountered in one taxon only (2n ¼

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

Downloaded from https://academic.oup.com/aob/article/100/3/505/165957 by Banasthali Vidyapith user on 10 September 2021

You might also like