Chromosome Botany (2009) 4: 83-86

© Copyright 2009 by the International Society of Chromosome Botany

Meiotic studies in some species of Pedicularis L. from cold desert regions of

Himachal Pradesh, India (North-West Himalaya)

Manjit Inder Singh Saggoo1 and Devendra Kumar Srivastava

Department of Botany, Punjabi University, Patiala 147 002, Punjab, India

Author for correspondence msaggoo@rediffmail.com

1

Received November 11, 2009; accepted December 11, 2009

ABSTRACT.　The chromosome numbers, meiotic course with pollen fertility and pollen size measurement of five species of
Pedicularis were assessed: P. gracilis, P. hoffmeisteri, P. pectinata spp. bipinnatifida, P. porrecta and P. punctata. All the five species
studied were diploid with 2n=16. The chromosome counts were the first report for P. pectinata spp. bipinnatifida, P. porrecta and P.
punctata. Also the phenomenon of cytomixis was reported in these three species. All the chromosome counts reinforced x = 8 as the
basic chromosome number for them.

KEYWORDS: Cytomixis, Pedicularis, Pollen fertility, Pollen size.

The high altitude location in and around Lahaul-Spiti area
of Himachal Pradesh (India) constitute the cold desert
region of North-West Himalaya. The area is quite rich
floristically (Aswal and Mehrotra 1994). Pedicularis L. is
considered to be the largest genus of the Scrophulariaceae
and also one of the largest genera of Angiosperms with
about 600-800 species primarily occurring in the arctic
and alpine regions of Northern Hemisphere (Wang and Li
2005; Mill 2001). At least half of the species of Pedicularis
occur in China mainly in the south west region especially
Hengduan Mountains region that expends up to south east
part of Tibet (Jie et al. 2004; Wang 2006). Nearly 100
species of the genus found their place in alpine Himalayan
belt (Garg and Husain 2003).
The species of Pedicularis are mostly annual with
perennial rootstocks and are hemi-parasites with short
sparsely branched roots. The flower has a characteristic
personate corolla of variable length which extends distally
into trilobed lower lip and an upper lip (galea). The name
of the genus Pedicularis, commonly known as ‘lousewort’,
finds its origin from Latin word “Pediculus” means louse,
as its decoction was used against lice on domestic animals
(Kriechbaum 2001). Presently an attempt has been made
to study the meiotic course, microsporogenesis and pollen
fertility in some of species of the genus Pedicularis from
the cold desert regions of North-West Himalaya.
MATERIALS AND METHODS
Plant materials for this study consisted of the wild taxa of
Pedicularis from Lahaul-Spiti and its adjoining areas of
Himachal Pradesh. Voucher specimens were deposited at
Herbarium, Department of Botany, Punjabi University,
Patiala (PUN). To obtain pollen mother cells (PMCs)
undergoing meiosis, young floral buds of individuals were
fixed in Carnoy’s fixative (6:3:1=absolute alcohol:
chloroform: glacial acetic acid v/v) for 24h at room
temperature and then were transferred to 70% alcohol and
stored under refrigeration until use. For meiotic studies
the anthers were squashed in 1% acetocarmine. A number
of slides were carefully examined per individual from
each species to determine chromosome numbers at
different stages and abnormalities were recorded. Pollen
stainability in glycerol-acetocarmine (1:1) was used to
estimate pollen viability. Measurements of diameter of
‘viable’ (i.e. stainable) pollen grains were done using
Ocular micrometer and Nikon microscope Eclipse 80i
system was used for photography.
RESULTS AND DISCUSSION
Presently, detailed meiotic studies were made on 15 taxa
belonging to five species of Pedicularis. The data regarded
locality, herbarium accession no. (PUN), meiotic chromosome
number and ploidy level of worked out species are provided
in Table 1.
　 Meiotic course in the individuals of Pedicularis gracilis
Wall. from Chamba area uniformly showed the presence
of eight bivalents at metaphase-I/II (Fig. I. 1) and normal
distribution at A-I. Some of the proximate PMC’s at
metaphase I (M-I) (13.20%) showed the phenomenon of
cytomixis (Fig. I. 2). Transfer of chromatin material from
one PMC to other was observed. Pollen fertility was
observed to be 89.33%. The present chromosome count
2n=16 in P. gracilis was in conformity with the earlier
reports for the species as given by Mehra and Gill (1968),
Gill (1972) and Amano (1999).
　 Three populations of P. hoffmeisteri Klotzsch. were studied
for the meiotic behaviour. The study revealed the presence
of eight bivalent at M-I (Fig. I. 3) in all the individuals
studied. Plants have high pollen fertility (94.52 ~ 96.11%).
The chromosome count 2n=16 was first report from Indian
part of the Himalayas and was in agreement with the 2n=16
as previously reported from Nepal (Amano 1999).
　 Pedicularis pectinata Wall. ex Benth. spp. bipinnatifida
Pennell. was occasionally found on grassy slopes in subalpine
regions of Lahaul-Spiti and Chamba areas. The presently
studied individuals from Trilokinath (2,760 m) area of
84 SAGGOO AND SRIVASTAVA

Lahaul-Spiti shows perfectly normal meiosis with 2n=16.
Plants collected from Gaurikund (3,920 m) area of Chamba
district showed 2n=16 (Fig. I. 4). Both of these populations
displayed the phenomenon of cytomixis (Fig. I. 5) in 9.10
and 11.02% of PMC’s respectively, resulting into reduced
pollen viability in both cases (Table 2). Plant individuals
from Gaurikund area was reported with average pollen
size of 25.50x24.63µm which had variable pollen size of
25.60x24.73 and 24.40x23.67µm. The present count of
2n=16 for ssp. bipinnatifida was first report for the taxa.
　 Study of meiotic course in P. porrecta Wall. ex Benth
showed the presence of 2n=16 (Fig. I. 6) and was quite
normal in the population of Rohtang (3,900 m) area. While
the individuals from its population in Trilokinath (2,650
m) area showed the phenomenon of cytomixis (Fig. I. 7) in
which 17.07% of PMC’s were involved and effect was
observed in the form of low pollen fertility (89.73%).
Variable size of pollens was also reported from the
population of Trilokinath area (Table 2). This was the first
report of chromosome number (2n=16) in the species.

Table 1. Locality and altitude, voucher specimen, meiotic chromosome number and ploidy level of wild plants of Pedicularis
Locality and altitude Voucher specimen Chromosome
Taxon Ploidy
(m) (PUN)＊ number (2n)
Pedicularis gracilis Wall. Chamba: Manimahesh, 16 2x
20978
Sundrasi (3,370 m)
P. hoffmeisteri Klotzsch. Lahaul-Spiti: 16 2x
51000, 51001
Koksar (3,500 m)
Manali: Gulaba (3,000 m) 49145, 49146 16 2x
Chamba : Manimahesh, 16 2x
Dunali (2,680 m) 50999
P. pectinata Wall. ex Benth. Lahaul-Spiti: 16 2x
50996
spp. bipinnatifida Pennell. Trilokinath (2,760 m)
Chamba: Manimahesh, 16 2x
Gaurikund (3,920 m) 50992
P. porrecta Wall. ex Benth. Manali: Rohtang (3,900 m) 49149, 49150 16 2x
Lahaul-Spiti: 16 2x
Trilokinath (2,650 m) 50995
P. punctata Decne Lahaul-Spiti: 16 2x
Koksar (3,500 m) 49151, 49152
Chamba: Manimahesh, 16 2x
Gaurikund (3,920 m) 50979
Sundrasi (3,370 m) 51003 16 2x
＊
PUN = Herbarium Code of Botany Department, Punjabi University, Patiala ( Holmgren and Keuken 1974).

Table 2. Meiotic abnormalities, pollen fertility and pollen size in Pedicularis species
Taxon Locality Abnormal Cytomixis Pollen Polen Size Rf.
PMCs with Fertility (µm)
L (%) B (%) Total No. of Meiotic (%)
PMC’s PMC’s stages
(%) involved
Pedicularis gracilis Sundrasi - 1.00 13.2 2-3 M-I 89.33 25.50x24.77 -
P. hoffmeisteri Koksar - - - - - 95.04 24.60x23.69 -
Gulaba - - - - - 96.11 24.47x23.72 -
Dunali - - - - - 94.52 24.60x23.50 -
P. pectinata Trilokinath 0.10 - 9.1 2-3 M-I 91.03 25.60x24.73 -
spp. bipinnatifida Gaurikund - - 11.02 2-3 P-I/M-I 89.27 25.50x25.50 0.10
24.40x23.67 0.90
P. porrecta Rohtang - - - - - 97.83 24.40x24.40 -
Trilokinath 1.00 - 17.07 2-3 M-I 89.73 25.80x25.80 0.11
24.50x24.50 0.89
P. punctata Koksar 0.70 - 3.04 2 M-I 94.21 24.40x24.40 -
Gaurikund 14.47 3.03 31.04 2-5 P-I/M-I 76.01 26.20x26.20 0.29
24.80x24.80 0.71
Sundrasi 11.33 1.01 19.37 2-3 M-I 79.80 26.10x26.10 0.19
25.10x25.10 0.81
PMC’s = pollen mother cells; L=Laggards; B = Bridges; P-I = Prophase-I; M-I = Metaphase-I; A-I = Anaphase-I; Rf .= Relative frequency
(Observed number of different size pollens/total number of fertile pollens).
 MEIOSIS IN PEDICULARIS FROM COLD DESERT REGIONS OF HIMACHAL PRADESH, INDIA 85

　 Meiotic study in P. punctata Decne. shows the presence
of eight bivalents at M-I (Fig. I. 8) in all the studied plants
belonging to different populations. The individuals from
Koksar (3,500 m) population show normal meiosis with
high (94.21%) pollen fertility. Meiotic course was
abnormal in other two populations. The phenomenon of
cytomixis involving inter PMC transfer of chromatin
material (Fig. I. 9 and 10) was observed in 31.04% and
19.37% of the observed PMCs in Gaurikund (3,920 m) and
Sundrasi (3,370 m) populations of the species respectively.
In these plants 2-5 PMCs at different stages of meiosis were
involved in cytomixis. The other abnormalities detected
were laggards (Fig. I. 11), bridges (Fig. I. 12), stickiness of
chromosomes, extra chromatin mass (Fig. I. 13), formation

Fig. I. Meiotic chromosomes in pollen mother cells (PMC’s) in Pedicularis. 1. PMC’s with 8:8 chromosome distribution
at metaphase II (MII) in P. gracilis. 2. PMC’s with cytomixis in P. gracilis. 3. A PMC with 8II at metaphase I (MI) in P.
hoffmeisteri. 4. A PMC with 8II at diakinesis in P. pectinata spp. bipinnatifida. 5. PMC’s involving in cytomixis in P.
pectinata spp. bipinnatifida. 6. A PMC with 8II at MI in P. pectinata spp. bipinnatifida. 7. A PMC showing partial transfer
of chromatin in P. pectinata spp. bipinnatifida. 8. A PMC with 8II at MI in P. punctata. 9. Group of three PMCs showing
cytomixis in P. punctata. 10. PMCs with double strand chromatin transfer in P. punctata. 11. A PMC with laggard at AI;
12. PMC with bridge at telophase II (TII) in P. punctata. 13. PMC at TII showing extra chromatin material in P. punctata.
14. Diyad in P. punctata. 15. Polyad in P. punctata. 16. Variable size fertile pollen in P. punctata. Bar=10µm.
86 SAGGOO AND SRIVASTAVA
of diads (Fig. I. 14), polyads (Fig. I. 15), etc. Cytomixis
and other abnormalities results into hypoploid, hyperploid
PMCs and consequently heterogenous size pollen grains
(Table 2). The pollen fertility was also low i.e. 76.01%
and 79.80% in the accessions of both the populations. The
present chromosome number count of n=8 is the first
report for the species.
　 Perusal of literature shows that genus Pedicularis is
based on x=6, 7 and 8. All the five presently investigated
species of the genus are diploids based on x=8. Three
species, namely P. pectinata Wall. ex Benth. spp.
bipinnatifida Pennell, P. porrecta Wall. ex Benth and P.
punctata Decne. have been investigated cytologically for
the first time. Some of the other Indian species of the
genus are based on x=6 and x=7. Verma and Dhillon
(1967) reported 2n=2x=12 for P. bifida (Buch.-Ham.)
Penn. (=P. carnosa Wall.) from India. Mehra and
Vasudevan (1972) reported x=7 as a base number for P.
bicornuta Klotz. The central Himalayan species of the
genus i.e. P. longiflora ssp. tubiformis and P. anserantha
var. elevatogaleata are also based on x=7 (Amano 1999,
Nakata et al. 2001). Intra-specific polyploidy based on x=8
has been reported in P. hoffmeisteri Klotzsch (2x, 4x) and
P. pauciflora Pennel (2x, 4x) from Nepal region (Amano
1999). Three other species, namely, P. brachyodonta
Schloss. and Vuk. (Strid and Franzen 1981), P. sceptrum-
carolinum L. (Krogulevich 1976) and P. villosa Ledeb. ex
Spreng. (Petrovsky and Zhukova 1981, Zhukova 1982) are
tetraploids based on the most common basic number x=8.
　 The phenomenon of cytomixis which was first recorded
in Crocus sativas L. by Kornicke (1901) and since then it
has been reported in wide range of flowering plants (Levan
1941, Sarvella 1958, Saggoo and Bir 1983, Heslop-Harrison
1966, Bione et al. 2000, Ghaffari 2006, Singhal and Kumar
2008). In the presently studied species the phenomenon of
cytomixis is seems to be natural. There is clear transfer of
chromatin material as well as chromosomes from one cell to
other causing serious problems in chromosomal segregations.
Unequal distribution of chromosomes may be responsible
for heterogeneous size pollen grains as reported earlier by
Singhal and Kumar (2008). The cytological aberration
observed in few populations may also be attributed to the
phenomenon of cytomixis. However, a good number of the
individuals among a population studied from the cold and
dry regions of Lahaul-Spiti and adjoining areas depict
regular meiotic course, normal microsporogenesis with
high pollen fertility.
ACKNOWLEDGMENTS. The authors are grateful to the
University Grants Commission, New Delhi for providing financial
assistance under the DRS SAP I and II and ASIST programmes.
Thanks are also due to Head, Department of Botany for necessary
laboratory facilities.
LITERATURE CITED
Amano, M. 1999. Cytotaxonomical study of Pedicularis
(Spermatophyta; Scrophulariaceae) in Annapurna Himal.
Nepal. Natural History Research. 5: 73-78.
Aswal, B. S. and Mehrotra, B. N. 1994. Flora of Lahaul-Spiti
( A Cold Desert in the North-West Himalaya). Bishen
Singh Mahender Pal Singh, India.
Bione, N. C. P., Pagliarini, M. S. and de Toledo, J. F. F. 2000.
Meiotic behaviour of several Brazilian soyabean varieties.
Genet. Mol. Biol. 23: 623-631.
Garg, A. and Husain, T. 2003. Strategies adopted by the alpine
genus Pedicularis L. (Scrophulariaceae) to overcome
environmental stress. Curr. Sci. 85: 10-12.
Ghaffari, G. M. 2006. Occurrence of diploid and polyploidy
microspores in Sorghum bicolor (Poaceae) is the result of
cytomixis. Afr. J. Biotechnol. 5: 1450-1453.
Gill, L. S. 1972. Chromosome numbers in West-Himalayan
bicarpellate species. II. Bull. Torrey Bot. Club. 99: 36-38.
Heslop-Harrison, J. 1966. Cytoplasmic connections between
angiosperm meiocytes; Ann. Bot. 30: 221-234.
Holmgren, P. K. and Keuken, W. (ed.) 1974. Index Herbariorum
Part-I. 1974 Reg. Veg. 92.
Jie, C., Hong, W., Zhijian, G., Mill, R. R. and Dezhu, L.
2004. Karyotypes of thirteen species of Pedicularis
(Orobanchaceae) from the Hengduan Mountain Region,
N. W. Yunnan, China. Caryologia. 57: 337-347.
Kriechbaum, K. 2001. Tibetan Medicinal Plant. CRC,
Medpharm Scientific Publ, pp. 85-97.
Kornicke, M. 1901. Uber ortsveranderung von Zellkarnern S B
Niederhein Ges Nature-U Heilkunde bonn A. pp.14-25.
Krogulevich, R.E. 1976. Chromosomne numbers of plant
species from the Tunkinsky Alpes (East Sayan). News
Sib. Dept. Acad. Sci. USSR, ser. Viol. 15: 46-52.
Levan, A. 1941. Syncyte formation in the pollen mother cells
of haploid Phleum pretense. Hereditas 27: 243-253.
Mehra, P.N. and Gill, L.S. 1968. In IOBP chromosome reports
XVI. Taxon 17: 199-204.
Mehra, P. N. and Vasudevan, K. N. 1972. In IOPB chromosome
number reports XXXVI. Taxon 21: 333-346.
Mill, R.R. 2001. Notes relating to the flora of Bhutan: XLIII.
Scrophulariaceae (Pedicularis). Edinburgh J.Bot. 58: 57-98.
Nakata, M., Takahashi, K. and Katoh, H. 2001. Cytological
studies on 31 alpine plants collected in Murodou-daira, mts.
Tateyama, central Japan. Bull. Bot. Gard. Toyama. 6: 5-20.
Petrovsky, V. V. and Zhukova, P. G. 1981. Chromosome
numbers and taxonomy of some plant species of Wrangel
Island. Bot. Zhurn. 66: 380-387.
Saggoo, M.I.S. and Bir, S.S. 1983. Cytomixis in certain
members of Acanthaceae and Labiatae from India. J.
Cytology and Genet. 18: 92-99.
Sarvella, P. 1958. Cytomixis and loss of chromosome in meiotic
and somatic cells of Gossypium. Cytologia. 23: 14-24.
Singhal, V. K. and Kumar, P. 2008. Impact of cytomixis on
meiosis, pollen viability and pollen size in wild population
of Himalayan poppy (Meconopsis aculeate Royle.).
Indian Academy of Science, J. Biosci. 33: 371-380.
Strid, A. and Franzen, 1981. In chromosome number reports
LXXII. Taxon. 30: 829-842.
Verma, S.C. and Dhillon, S.S. 1967. In IOPB chromosome
number reports XI. Taxon. 16: 215-222.
Wang, H. 2006. Pedicularis L. in Flora Yunnanica. Wu, Z. Y.,
Beijing Science Press. pp. 468-611.
Wang, H. and D. Z. Li. 2005. Pollination biology of four
Pedicularis species (Scrophulariaceae) in northwestern
Yunnan, China. Ann. Missouri Bot. Gard. 92:127-138.
Zhukova, P. G.1982. Chromosome numbers of some plant
species of north-eastern Asia. Botaniceskjij Žurnal SSSR.
67: 360-365