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© 2011 The Japan Mendel Society Cytologia 76(3): 1–6

Meiotic Studies in 14 Species of the Nepeta L. (Lamiaceae)
From Cold Desert Regions of Lahaul-Spiti and Adjoining Areas
of Northwest-Himalaya, India
Manjit Inder Singh Saggoo, Devendra Kumar Srivastava*
and Punam Grewal
Department of Botany, Punjabi University, Patiala 147 002, Punjab, India

Received November 23, 2010, accepted January 12, 2011

Summary Chromosome numbers, meiotic course and pollen viability were given for 14 species
of genus Nepeta L. collected from different localities in Lahaul-Spiti, Kulu, Chamba, Kinnour and
Sirmour districts of Himachal Pradesh, India. Except for N. leucophylla (2n=4x=36, tetraploid), all
the other 13 species (N. campestris, N. connata, N. discolor, N. eriostachya, N. elliptica, N. govani-
ana, N. graciliflora, N. leucolaena, N. linearis, N. longibracteata, N. podostachys, N. spicata and N.
supina) were diploid (2n=2x=18). Analysis of result shows that the majority of Indian Nepeta spe-
cies are present with x=9.

Key words Chromosome number, Indian Nepeta L., Meiotis, Pollen viability.

Nepeta L. (Lamiaceae), known well as catmint or blue ice, are annual or perennial herbs dis-
tributed in temperate Europe, Asia, North Africa and the mountains of tropical Africa. The genus
is the largest genus of the tribe Nepetae, comprising about 250 species (Mabberley 1997). Nearly
35 species of the genus are found in the Western Himalayan belt of India, extending throughout the
Himalayas from Kashmir to Sikkim (Hooker 1885). The species are traditionally used as antiasth-
matic, diuretic, diaphoretic, febrifuge and sedative agents (Chopra et al. 1956). Alcoholic extract of
N. hindostana (Roth.) Haines. is reported to lower the blood cholesterol (Said 1969). Many species
have been investigated for their essential oils and medicinally important compounds (Bottini et al.
1992, Kishore and Dwivedi 1992, Baser et al. 2000, Sonboli et al. 2005). Despite this, informa-
tion on chromosome numbers in Indian species of Nepeta is incomplete. The present study aims at
a compilation of our knowledge of the cytology of the genus from Lahoul-Spiti and the adjoining
high altitude regions of the Himalayas.

Materials and methods

Plant materials for cytological studies were collected during the months of June–August in
2007–2010 from Lahaul-Spiti and adjoining high altitude areas of Himachal Pradesh. The plants
were identified by consulting different floras of the western Himalayan region (Hooker 1885,
Aswal and Mehrotra 1994) and were confirmed with authentic specimens lodged in the herbarium
of Botanical Survey of India, Dehradun, India. All specimens were deposited in the herbarium,
Department of Botany, Punjabi University, Patiala, India (PUN).
Floral buds of suitable sizes were fixed in Carnoy s fixative (6 : 3 : 1=Absolute ethanol : chloro-
form : glacial acetic acid, v/v) for 24 h and preserved in 70% ethanol at 4 C. Anthers were squashed
in 1% acetocarmine. More than 2000 pollen mother cells (PMCs), including microsporocyte in

* Corresponding author, e-mail: devsrivastv@gmail.com
2 M. I. S. Saggoo et al. Cytologia 76(3)

Table 1. Data showing locality with voucher specimen (PUN), 2n chromosome count/Ploidy and Pervious
reports with remarks of different species of Nepeta L.

Pollen
Taxon Locality (PUN)* 2n Previous reports/Remarks Viability
%

N. campestris Benth. Kullu: Manali, 2,600 m (49049). 18 Present study 96.1
N. connata Royle ex Benth. Pangi: Shali, 2,600 m (54183) 18 Present study 97.0
Killar, 2,650 m (54182). 18
N. discolor Royle ex Benth. Lahaul: Gramphoo, 3,800 m (49021) 18 Present study 952
Koksar, 3,250 m (54163) 18 67.5
Spiti: Kye, 4,000 m (54172) 18 95.7
Losar, 3,960 m (54158). 18 94.6
N. eriostachya Benth. Lahaul: Trilokinath, 2,760 m (54147) 18 Saggoo 1983, Saggoo & 96.5
Darcha, 3,300 m (54148). 18 Bir 1983, Gill 1984.
N. elliptica Royle ex Benth. Sirmour: Churdhar, 2,650 m (54185). 18 Gill 1984. 97.3
N. govaniana Benth. Kullu: Manali, Gulaba, 3,600 m (54167) 18 Gill 1984, Bir & Saggoo 93.7
Manikaran,Malana,2,600m (54180). 18 1984.
N. graciliflora Benth. Lahaul: Udaipur, 2,900 m (54184). 18 Gill 1984, Saggoo & Bir 95.5
1983.
N. leucolaena Benth. ex Pangi: Shol, 3,000 m (54178) 18 Khatoon & Ali, 1993/First 86.1
Hook. f. Kinnour: Nako, 3,600 m (54179). 18 report to study area.
N. leucophylla Benth. Lahaul: Keylon, 3,677 m (54150) 36** Gill 1984, Saggoo 1983. 91.5
Chamba: Manimahesh, 3,320 m (54152). 36
N. linearis Royle ex Benth. Lahaul: Udaipur, 2,740 m (54170) 18 Gill 1984; Jee et al. 1989. 96.3
Pangi: Shali, 3,300 m (54176). 18
N. longibracteata Benth. Lahaul: Keylong, 3,677 m (49023). 18 Present study 92.0
N. podostachys Benth. Lahaul: Tandi, 3,400 m (49040). 18 Podlech & Dieterle 1969; 93.1
Budantsev et al. 1992/
First report to study area.
N. spicata Benth. Lahaul: Trilokinath, 2,700 m (54161) 18 Gill 1984, Khatoon & Ali 91.0
Spiti: Tiling , 3,400 m (54157). 18 1993.
N. supina Stev. Kullu: Manali, Rohtang Pass, 3,900 m 18 Present study 89.3
(49051).

* PUN=Accession no. Herbarium, Department of Botany, Punjabi University, Patiala (Holmgren and Keuken 1974)
** Tetraploid, rest of all taxa are diploid.

meiosis I/II, and meiotic products were analyzed in each population. Pollen stainability in glycerol–
acetocarmine (1 : 1) was used to estimate pollen viability. A measurement of the diameter of viable
pollens was done using Ocular micrometer and for photography a Nikon microscope Eclipse 80i
system was used.

Results and discussion

Presently, detailed meiotic course was analyzed in 24 populations belonging to 14 species of
Nepeta. The species were collected from different localities in Lahoul-Spiti, Kulu, Chamba, Kin-
nour and Sirmour districts of Himachal Pradesh. The data regarding locality, herbarium accession
no. (PUN), meiotic chromosome number (2n), previous reports and pollen viability of worked out
species are provided in Table 1.

N. campestris Benth.
Plants for meiotic analysis were collected from the population at Manali (PUN49049). Several
2011 Meiotic Studies in Himalayan Nepeta L. 3

Fig. 1. Meiotic chromosome count in 14 Indian species of Nepeta. a; N. campestris: A PMC showing 9II at
M-I, b; N. Connata: A PMC with 9II at M-I, c; N. discolor: A PMC with 9 : 9 distribution at A-I, d;
N. eriostachya: A PMC with 9II at M-I, e; N. elliptica: A PMC showing 9II at M-I, f; N. govaniana:
A PMC with 9 bivalents at M-I, g; N. graciliflora: A PMC with 9 : 9 distribution at A-I, h; N. leu-
colaena: A PMC with 9II at diakinesis. i-j; N. leucophylla: I; A PMC showing 18II at diakinesisj. A
PMC with 9II at M-I, k; N. linearis: A PMC showing 18I at late M-I, l; N. longibracteata: A PMC
with 9II at diakinesis, m; N. podostachys: A PMC having 9II at diakinesis., n; N. spicata: A PMC
showing 9 : 9 distribution at A-I, o. N. supina: A PMC with 9 : 9 distribution at A-I. Bar=10 μm.

PMCs showed n=9 (Fig. 1a), which is the first chromosome count for the species. Meiotic course
was normal with high pollen viability (96.1%).
4 M. I. S. Saggoo et al. Cytologia 76(3)

Fig. 2. Meiotic aberrations in N. discolor from Koksar. A; A PMC with 9II at M-I, b; A PMC showing
chromosome stickiness at M-I, c; A PMC showing Stickiness at A-I, d; Two PMCs showing cyto-
mixis, e; A group of 3 PMCs involving in chromatin transfer, f; Two PMCs with bridge (arrowed)
at A-I, g; Two PMCs showing bridge and laggard (arrowed) at A-I, h; Heterogeneous size of fertile
pollens, I; A group of 4 normal size fertile pollens. Bar=10 μm.

N. connata Royle ex Benth.
The species is rare and plants were collected from the Shali (PUN54183) and Killar
(PUN54182) villages of Pangi valley in the Chamba district. The meiotic course was normal with
n=9 (Fig. 1b) and plants showed high pollen viability (97.0%). The chromosome count in this spe-
cies is the first report for the species.

N. discolor Royle ex Benth.
Plants for meiotic study were collected from 4 populations from Lahoul-Spiti, i.e. Gramphoo
(PUN49021), Koksar (PUN54163), Key (PUN54172) and Losar (PUN54158). All the plants are
diploid with chromosome number 2n=2x=18 (Fig. 1c), which is also the first report for the spe-
cies. Meiotic course in these populations was normal except in the Koksar population (Fig. 2a),
where chromosome stickiness (Fig. 2b, c) and cytomixis were observed with inter-PMCs chromatin
transfer in about 30% PMCs (Fig. 2d, e). Bridges and laggards (17.0% of PMCs) were present at
anaphase-I (A-I) (Fig. 2f, g). The consequences of these meiotic abnormalities were observed in the
form of low pollen viability (67.5%) and heterogeneous size of pollen grains ranging from 30.43 to
37.40 μm (Fig. 2h, i).

N. leucolaena Benth. ex Hook. f.
The chromosome number n=9 (Fig. 1h) in the present species from the populations at Shol
2011 Meiotic Studies in Himalayan Nepeta L. 5

(PUN54178) and Nako (PUN54179) was in agreement with the earlier reports of 2n=18 in Pakistan
(Khatoon and Ali 1993). The plants showed normal meiotic courses with 86.1% apparently viable
pollens. The chromosome number is the first chromosome count in Indian material.

N. longibracteata Benth.
Individuals of the species were collected from the Keylong population (PUN49023) and
showed chromosome number of n=9 (Fig. 1l), the first record for the species. About 92% of pollens
were viable.

N. podostachys Benth.
PMCs of the individuals from Tandi (PUN49040) were in conformity with the earlier reports
of 2n=18 (Fig. 1m) for the present species (Podlech and Dieterle 1969, Budantsev et al. 1992). Nor-
mal meiotic course and pollen fertility (93.1%) were observed. The 2n=18 in this species is the first
chromosome count in India.

N. supina Stev.
Plant materials were collected from Rohtang Pass (PUN49051). PMCs showed n=x=9 (Fig.
1o) and this number is the first report for the species. Meiosis proceeded normally resulting in
89.3% pollen viability.
In rest of the species, viz. N. eriostachya, N. elliptica, N. govaniana, N. graciliflora, N. leuco-
phylla, N. linearis and N. spicata, the chromosome numbers observed confirmed previous reports
(Table 1).
Perusal of literature shows that chromosome number information for 77 species of Nepeta L.
is available and that the genus had basic chromosome numbers, x=7, 8, 9, and 13. Among these
base numbers, the majority are based on x=9 (57 species) followed by x=8 (11 species). The chro-
mosome numbers varies from 2n=14 to 2n=54 with polyploidy cytotypes reported in 15 species,
mostly tetraploids. One species, N. beltranii Pau., is at hexaploid level with 2n=54 (Ubera 1983).
Intra-specific polyploidy with 2n=18, 36 is reported in N. hindostana Haines (Saggoo 1983), N.
mollis Benth. (Saggoo and Bir 1983, Gill 1984) and N. subsessilis Maxim. (Aryavand 1975, Nakata
et al. 2001), while in N. camphorata Boiss. it is based on x=8 (2n=16: Baden 1983, 2n=32: Gill
1981). Taking into consideration the available cytological data, it has been observed aneuploidy is
operative both at diploid and polyploidy levels. Five species, namely N. cataria L. (2n=34, 36), N.
distans Royle. ex Benth. (2n=18, 26), N. grandiflora M. Bieb. (2n=34, 36), N. nepetella L. (2n=34,
36) and N. transcaucasica Grossh. (2n=16, 18), show intra-specific aneuploidy without effecting
ploidy level.
All the presently studied species except N. leucophylla Benth. (4x) are diploid based on x=9.
Cytologically these species have normal meiotic behavior with high pollen viability. Low pollen
viability and heterogenous size of pollens in one of the population of N. discolor Royle ex Benth.
from the Koksar population (PUN54163) reflects the role of meiotic abnormalities especially lag-
gards, bridges stickiness of chromosomes and cytomixis as suggested by Singhal et al. (2008) and
Fatemeh et al. (2010). However, the influence of environmental factors (Jones and Newell 1948,
Kurtz and Liverman 1958), nutrition (Bell 1959), ploidy (Gould 1957, Kapadia and Gould 1964)
and geographical variation (Cain and Cain 1944) on pollen size and fertility in flowering plants
should not be underestimated.

Acknowledgments

The authors are thankful to University Grant Commission, New Delhi for providing financial
assistance under the DRS SAP III and ASIST programmes. Thanks are also due to the Head of the
6 M. I. S. Saggoo et al. Cytologia 76(3)

Department of Botany, Punjabi University, Patiala for providing the necessary laboratory facilities.

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