‘INSIGHT’ an International Journal of Science(ISSN: 2394-0360); Volume 1 2014

Abnormal Meiosis in Tetraploid (4x) Clinopodium umbrosum (M. Bieb) Koch from
Lahul Valley of Himachal Pradesh, India
*1
Devendra Kumar Srivastava and 2Manjit Inder Singh Saggoo
1
Department of Biology, D.A.V. College, Hoshiarpur-146001, Punjab (India);
2
Department of Botany, Punjabi University, Patiala-147002, Punjab, India);
(*Corresponding author: devsrivastv@gmail.com)
(Received: Oct-19, 2014; Revised: Nov-29, 2014; Accepted: Dec-07, 2014)
ABSTRACT
Present paper deals with the chromosome numbers, meiotic course and pollen viability of
Clinopodium umbrosum (M. Bieb) Koch which was assessed cytologically in six populations growing in
different localities of Lahaul valley and its adjoining areas of Himachal Pradesh (H.P.) viz. Hole
(3,500m), Trilokinath (2,760m), Keylong (3,350m), Udaipur (2,600m), Tandi (3,200m) and Zero-point
(2,600m). Except, the population growing in Trilokinath (2n=4x=20), all the other populations were
reported with normal meiotic course and microsporogenesis. It was present with chromosome stickiness
(23.25%), bridges (12.60%), laggards (17.52%), cytomixis (21.33%), diads (08.50%), triads (06.33%)
and polyads (09.50%) with or without micronuclei and unequal daughter cells in plants of Trilokinath
population. The consequences of these meiotic abnormalities were in the form of heterogenous sized
pollen grains and low (68.33%) pollen viability.
Key words: Clinopodium umbrosum, Chromosome number, Meiotic course and Population.

INTRODUCTION Bieb) Koch.] is commonly known as Birchee

The genus name Clinopodium is derived
from the Greek word klinopodion, the klino means
‘a bed’ and the podion means ‘a little foot’, which
some observers say refers to how the plants
terminal clusters resemble the knobs on the foot of
a bed frame (Quattrocchi 2000). The genus is
closely allied to Calamintha and many species are
taxonomically ill defined. About 271 species,
mainly distributed in Europe and Asia, are recorded
in literature and approximately 52.4% (142) of
them are taxonomically accepted (Srivastava 2012).
In Indian floras, total number of the species for the
genus Clinopodium is not fully described yet.
However, nearly six species described under the
name of Calamintha are distributed throughout
temperate Himalayas up to 3,600m. Most of the
plant species are used as medicinal herbs. Species
Clinopodium umbrosum (M. Bieb) Kuntze [=Syn.
Calamintha umbrosa Benth., C. umbrosa (M. Bieb)
Fisch. & C. A. Mey., Clinopodium umbrosum (M.
(Hindi) and easily available medicinal plant of
Lahaul valley. It is mainly distributed in temperate
Himalayas from Kashmir to Bhutan at an altitude of
1,200 to 3,600m. Here, it is found on moist slopes.
Species is native to India and whole plant is used
by the local people as astringent, carminative or
heart tonic. The leaves and roots possess
camphoraceous properties, also yield an essential
oil (Srivastava 2012). Locally leaf powder of C.
umbrosum is used as antiseptic, for stomach-ache
and liver disease (Srivastava 2012). The present
study aims at a compilation of our cytological
knowledge based on population of the species from
Lahaul valley and its adjoining high altitude regions
of the Himalayas.
MATERIAL AND METHODS
Materials for meiotic study were collected
during the months of June–August. For the meiotic
preparations appropriate sized of young floral buds

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 ‘INSIGHT’ an International Journal of Science(ISSN: 2394-0360); Volume 1
growing in their natural habitats were fixed in
Carnoy`s fixative (6:3:1=Absolute ethanol:
chloroform: glacial acetic acid, v/v) for 24 h and
preserved in 70% ethanol at 4 0C. Anthers were
squashed in 1% acetocarmine. More than 2000
pollen mother cells (PMCs), including
microsporocyte in meiosis I/II, and meiotic
products were analyzed in each population. Pollen
stainability in glycerol–acetocarmine (1:1) was
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used to calculate pollen viability. The diameter
measurement of viable pollen grains was done
using ocular micrometer and for photography a
Nikon microscope Eclipse 80i digital system was
used. The plants were identified by consulting
different floras of the western Himalayan region
(Hooker 1884; Aswal and Mehrotra 1994) and were
confirmed with authentic specimens lodged in the
herbarium of Botanical Survey of India, Dehradun,
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 ‘INSIGHT’ an International Journal of Science(ISSN: 2394-0360); Volume 1
India. All specimens were deposited in the
herbarium, Department of Botany, Punjabi
University, Patiala, India (PUN).
RESULTS
Presently detailed meiotic studies were
made on six taxa belonging to the species
Clinopodium umbrosum. The taxa were collected
from the different populations (viz. Hole 3,500m,
Trilokinath 2,760m, Keylong 3,350m, Udaipur
2,600m, Tandi 3,200m and Zero-point 2,600m) in
the high altitude localities of Lahaul-Spiti. The data
regarding localities herbarium accession number
(PUN), meiotic chromosome number (2n) pollen
viability percentage (PV%) and previous reports of
worked out taxa are provided in Table 1.
Following the cytological study on the
collected taxa from different populations revealed
almost similar chromosome count of n=10 (Fig.
1a). Meiotic course in almost five population was
normal (Table 2), where, less than 5% proximate
pollen mother cells (PMC’s) were observed with
the phenomenon of chromosome stickiness and
cytomixis. Pollen fertility was high in these
populations and observed to be above 90% (see
Table 1 & 2). Only the result of the population with
present cytological interest is described here.
Meiotic observations in the floral buds,
collected from the plant population growing in
Trilokinath area (2,760m) of Lahaul valley were
reported with chromosome count of n=10 (Fig.1b-
c). Meiotic course in this population was differing
from that of populations collected from the other
sites (Table 1 and 2). At M-I nearly 23.25% of
PMC’s were observed with chromosome stickiness
(Fig.d). PMCs with multiple bridges (12.60%) and
laggards (17.52%) were present at anaphase-I (A-I)
(Fig.1e-f). Approximately 21.33% of PMCs were
present with inter PMC’s chromatin transfer i.e.
cytomixis (Fig.1g). Microsporogenesis was obser-
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-ved with diads (8.50%), triads (6.33%) and
polyads (09.50%) along with or without
micronuclei and unequal daughter cells (Fig.1h-j).
The consequences of these meiotic abnormalities
were observed in the form of heterogenous sized
pollen grains and low (68.33%) pollen viability
(Fig.1k-l). Heterogenous sized pollen grains were
studied in three categories namely, P (small sized
pollens; falls in averaged size ranged of x; where x
≤ 27.82 x 25.83 μm), Q (normal medium sized
pollens; falls in averaged size ranged of y; where
27.82 x 25.83 μm < y ≤ 34.75 x 32.89 μm) and R
(large sized pollens; falls in averaged size ranged of
z; where z ˃ 34.75 x 32.89 μm). Pollen grains
falling in range of y (category Q) and z (category R)
were observed in all the collected populations, only
the population of Trilokinath area was observed
with pollens falling in all the ranges of x, y and z
i.e. categories P, Q and R. The relative frequency %
(Rf-value) of medium (category Q) sized (T; Table
2) fertile pollen grain was high in almost all the
populations while it was lowest (Rf %=55.98) in
Trilokinath population (PUN54963) and highest (Rf
%=97.84) in the population (PUN54945) of Hole
area of Chamba district (Table 2).
DISCUSSION
The present chromosome count of n=10
confirms the earlier record of 2n=20 by Mehra and
Gill (1968) and by Gill (1971 and 1984) from
different parts of India (Table 1).
Chromosomally, 27 species (including 6
subspecies) of the genus are known. Various
members exhibit a wide range of chromosome
numbers as 2n=10, 16, 18, 20, 22, 24, 30, 32, 36,
38, 40, 42, 48 and 72, which indicate its polybasic
nature with base numbers of x=5, 8, 9, 10, 11, 12,
15 and 19 (Table-3). Presence of x=5 (2n=10) in
the genus reported by Cherian and Kuriachan
(1990). Gill (1984) and Singh (1995), however,
suggested the x=10 as the primary and the probable
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 ‘INSIGHT’ an International Journal of Science(ISSN: 2394-0360); Volume 1
ancestral base number. Intra-specific polyploidy is
exhibited by three species based on x=10. With
discovery of x=5 the base number x=10 seem to be
secondary in origin.
It is observed that, in India, the genus exists
with base numbers x=5, 8, 9 and x=10. Species C.
umbrosum is reported with three base numbers x=5,
8 and 9. Whereas species C. vulgare L. is present
with 4x and 8x based on x=5. Based on this the
presently investigated taxa [Clinopodium
umbrosum (M. Bieb) Kuntze; 2n=20] was existed at
tetraploid (4x) level.
Cytologically all the individuals of the
population have normal meiotic behavior with high
(>90%) pollen viability. Presently, the irregularities
and disturbances in meiosis contributed to the
degenerative processes of pollen development in
tetraploid (4x) population of Trilokinath area and
resulted in low (<70%) viability. The low pollen
viability in this population reflects the role of
meiotic abnormalities especially laggards, bridges
and stickiness of chromosomes as suggested in
other species by Saggoo and Srivastava (2009),
Srivastava (2012), Kumar et al. (2013) and Kumar
and Dwivedi (2013). Further, presence of
heterogenous size pollen grains and high frequency
of large sized pollen grains in the population may
be partially due to meiotic irregularities and
partially due to cytomixis which are responsible for
unequal chromatin distribution in these pollens.
Similar observation and concept of cytomixis was
made in other flowering plants by different workers
(Saggoo and Srivastava 2009, Kalinka et al. 2010,
Saggoo et al. 2011, Srivastava 2012, Kravets 2013).
Now it is well known that cytomixis is a natural
and normally genetically controlled phenomenon.
But 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
2014
and Cain 1944) on pollen size and fertility in
flowering plants could not be underestimated.
REFERENCES
Aswal BS and Mehrotra BN (1994). Flora of
Lahaul-Spiti (A Cold Desert in the North-
West Himalaya). Bishen Singh Mahender
Pal Singh, India. pp. 466-522.
Bell CR (1959). Mineral nutrition and flower to
flower pollen size variation. Am. J. Bot. 46:
621–624.
Bir SS and Saggoo MIS (1985). Cytological
studies on members of family Labiatae
from Kodaikanal and adjoining areas
(South India). Proc. Indian Acad. Sci.
(Plants) 94:619-626.
Cain SA and Cain LG (1944). Size frequency
studies of Pinus palustrius pollen. Ecology
25: 229–232.
Cherian M and Kuriachan PI (1990). A new
chromosome count in Calamintha umbrosa
Benth. (Labiatae) and its bearing on the
evolution of the genus. Sci. & Cult. 56:
133–134.
Gill LS (1971). Cytology of West-Himalayan
Labiatae: Tribe Satureieneae. Bull. Bot.
Soc. Bengal, 24:203-207.
Gill LS (1984). The incidence of polyploidy in the
West-Himalayan Labiatae. Rev. Cytol. Biol.
Veget. Bot. 7: 5-16.
Gould FW (1957). Pollen size as related to
polyploidy and speciation in the
Andropogon saccharoides - A. barbinoidis
complex. Brittonia, 9: 71–75.
47
 ‘INSIGHT’ an International Journal of Science(ISSN: 2394-0360); Volume 1
Holmgren PK and Keuken W (1974). Index
Herbariorum Part-I. Reg. pp. 92.
Hooker JD (1884). Flora of British India-Reeve, L.
and CO., Ltd., Kent., Vol. IV.
Jones MD and Newell LC (1948). Size, variability
and identification of grass pollen. J. Am.
Soc. Agron. 40: 136–138.
Kalinka A, Achrem M, Rogalska SM (2010).
Cytomixis-like chromosomes/chromatin
elimination from pollen mother cells
(PMCs) in wheat-rye allopolyploids. The
Nucleus, 53(3):69-83.
Kapadia ZJ and Gould FW (1964). Biosystematics
studies in the Bouteloua curtipendula
complex III. Pollen size as related to
chromosome numbers. Am. J. Bot. 51: 166–
172.
Khatoon S (1991). Polyploidy in the flora of
Pakistan – an analytical study. Ph. D.
thesis, University of Karachi, Pakistan.
Khatoon S and Ali SI (1993). Chromosome Atlas of
the Angiosperms of Pakistan. Department
of Botany, University of Karachi, Karachi.
Kravets EA (2013). Cytomixis and its role in the
regulation of plant fertility. Russian J.
Devlopmental Biology. 44(3): 113-128.
Krishnappa DG and Basavaraj I (1982). In IOPB
chromosome number reports LXXV. Taxon
31: 361–362.
Kumar G and Dwivedi K (2013). Cytogenetical
evidences of abnormal meiosis and 2n
pollen formation via colchicines in
microsporogenesis of Brassica campestris
L. Int. J. Rres. Plant Sci. 3(2):18-24.
2014
Kumar P, Rana PK, Himshikha, Singhal VK and
Gupta RC (2013). Cytogeography and
phenomenon of cytomixis in Silene
vulgaris from cold regions of Northwest
Himalayas (India). Plant Syst. Evol. DOI
10.1007/s00606-013-0922-7.
Kurtz EB and Liverman JL (1958). Some effects of
temperature on pollen characters. Bull.
Torrey Bot. Club, 85:136–138.
Mehra PN and Gill LS (1968). In IOPB
chromosome number reports XVI. Taxon.
17:199-204.
Pal S (1971). Cytotaxonomy of Labiatae. Proc.
48th Indian Sci. Congr., 3:297.
Quattrocchi U (2000). CRC World Dictionary of
Plant Names. Vol.- I, CRC Press: Boca
Raton; New York; Washington,DC; USA.
London, UK. pp.91.
Saggoo MIS (1983). Cytomorphological studies on
plants of economic importance of
Bicarpellatae from India. Thesis. Punjabi
University Patiala, India. p.1-259.
Saggoo MIS and Bir SS (1981). In IOPB
chromosome number reports LXXI. Taxon
30: 515.
Saggoo MIS and Bir SS (1983). Chromosome
number of certain Acanthaceae and
Labiatae VI. SOCGI Plant Chromosome
number report. I. J. Cytol. Genet. 18: 56-
63.
Saggoo MIS and Bir SS (1986). Meiotic studies on
some East Himalayan members of family
Labiatae. J. Indian Bot. Soc. 65: 304-309.
48
 ‘INSIGHT’ an International Journal of Science(ISSN: 2394-0360); Volume 1 2014

Saggoo MIS and Srivastava DK (2009). Meiotic

studies in some species of Pedicularis L.
from cold desert regions of Himachal
Pradesh, India (N.W. Himalaya). Chomo.
Bot. 4:83-86.

Saggoo MIS, Srivastava DK and Grewal P (2011).

Meiotic studies in 14 species of the Nepeta
L. (Lamiaceae) from cold desert regions of
Lahaul-Spiti and adjoining areas of
Northwest-Himalaya, India. Cytologia.
76(3): 231-236.

Singh TP (1995). Alteration in the basic

chromosome numbers as a means of
speciation in Labiatae. Feeders Repr.
106(1-2): 39-47.

Srivastava DK (2012). Cyto-morphological

diversity in species of Labiatae and
Scrophulariaceae from Lahaul-Spiti and
adjoining areas. Thesis. Dept. of Bot.
Punjabi University Patiala.

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