Chromosome Botany (2013) 8: 41-44

© Copyright 2013 by the International Society of Chromosome Botany

Chromosome numbers in few members of Scrophularia L. from cold desert

regions of Lahaul-Spiti, H. P., India

Devendra Kumar Srivastava1 and Manjit Inder Singh Saggoo

Department of Botany, Punjabi University Patiala 147002, India

1
Author for correspondence: (devsrivastv@gmail.com)
Received January 1, 2013; accepted February 13, 2013

ABSTRACT.　Chromosome numbers, ploidy and pollen fertility of the six species of Scrophularia L. were assessed from cold
desert regions of Lahaul-Spiti: Scrophularia calycina Benth. (2n=26), S. decomposita Royle ex Benth. (2n=24), S. edgeworthii
Benth. (2n=48), S. himalensis Royle ex Benth. (2n=24), S. koelzii Penn. (2n=20), S. polyantha Royle ex Benth (2n=24+2-3B).
Meiotic chromosome count in S. decomposita was varied report while in S. koelzii Penn. it was recorded for the first time. Except
S. edgeworthii (4x) rest all the species were diploid (2x).

KEY WORDS: Chromosome numbers, Lahaul Spiti, Scrophularia L.

Scrophularia L. is one of the important member of the
Scrophulariaceae and commonly known as the figworts.
The genus is represent with about 200 polymorphic
species (Fischer 2004) and distributed in both the
hemispheres in Europe, Southeast Asia, Northern parts of
America and Mountains of Indonesia to New Zealand.
Nearly 13 species of the genus found their place in alpine
Himalayan belt of India (Dhillon 1979; Hooker 1885).
The genus owes its name to the fact that it is reputed to be
a remedy against Scrofula, a disease of glands (Qaiser et
al. 1988). Many species belonging to this genus have been
used as folk remedies science ancient times (Heather and
Henderson 1994; Galindez et al. 2002).
　 For the present genus literature shows that the earlier
workers have conducted cytological surveys mainly from
North India, Central and South India (Kusumlata 2006).
From the Cold desert regions of the Lahaul-Spiti most of
the scientific work limited up to the taxonomic
characterization and little attention has been given on the
cytology. Area is situated between 31°44′57″-32°59′57″ N
latitudes and 76°46′29″-78°41′34″ E longitude in Himachal
Pradesh. Cytologically very few reports are available
from this area in the Scrophulariaceae (Saggoo and
Srivastava 2009). The present work is needful effort to fill
the gap of cytological knowledge in members of the
Scrophulariaceae from the cold and harsh climatic
conditions of Lahaul-Spiti.
MATERIALS AND METHODS
Materials for meiotic study were collected from natural
fields during the months of June to August. For the meiotic
preparations appropriate sized floral buds growing in their
natural habitats were flxed in Carnoy’s flxative (6:3:1=
Absolute ethanol : chloroform : 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
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.
　 The plants were identifled by consulting different
floras of the western Himalayan region (Hooker 1885;
Aswal and Mehrotra 1994) and were conflrmed 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).
RESULT AND DISCUSSION
The present study includes a detailed meiotic analysis in
six species of Scrophularia. The data regarding locality,
herbarium accession no. (PUN), meiotic chromosome
number (2n), the previous reports and pollen viability of
worked out species are provided in Table 1.
S. calycina Benth.　Accession for meiotic study was
collected from Rudarnar (2,650m) population. It reveals
the presence of 13:13 chromosome distribution at anaphase
I (Fig. 1a). Meiotically plant accession was normal and was
recorded with high pollen fertility (98.50%). The present
chromosome count of 2n=26 in S. calycina Benth. is
similar to the report of Gill (1971) from the Western
Himalaya.
S. decomposita Royle ex Benth. (=S. lucida Hook. f.,
S. griffithii Benth.)　For the meiotic analysis the specimens
were collected from different localities which represents
the different populations (Table 1). Meiotic preparations
in these populations revealed similar chromosome count
of 2n=24 at metaphaseI and diakinesis (Fig. 1b-c). Nearly
7.05% of PMC’s in the population of Shali Village were
42 SRIVASTAVA AND SAGGOO

Table 1. Data on locality with altitude, accession number (PUN), meiotic chromosome count (2n), ploidy level, pollen
fertility (%) and previous reports on cytologically investigated species of Scrophularia L.
*
Taxa/ Locality PUN 2n PF (%) Ploidy Previous Reports (2n) Remarks
S. calycina Benth.
/Khirganga: Rudarnar (2,650m) 55015 26 98.50 2x 26: Gill 1971. Previous report of 2n= 26 confirm.
S. decomposita Royle ex Benth.
/Chamba: Manimahesh, Sundrasi (3,380m) 54942 24 95.50 2x 26: Vasudevan 1975. Varied report
/Spiti: Key (3,750m) 54948 24 99.01 2x
/Spiti: Kibber (3,800m) 54985 24 91.25 2x
/Chamba: Pangi Valley, Shali (2,700m) 54986 24 69.33 2x
/Lahaul: Keylong, (3,250m) 54989 24 100 2x
/Spiti: Kaza (3600m) 55044 24 97.50 2x
/Lahaul: Keylong, Shasur (3,000m) 54990 24 100 2x
/Spiti: Pin Valley, Mudh (3,300m) 54992 24 100 2x
S. edgeworthii Benth.
/Parvati valley: Malana (2,650m) 54988 48 98.50 4x 48: Mehra and Vasudevan 1972. Previous report of 2n= 48 confirm.
/Lahaul: Udaipur, Jhalma (2,700m) 54991 48 97.66 4x
S. himalensis Royle ex Benth.
/Lahaul: Udaipur, Jahalma (2,650m) 55017 24 100 2x 20: Mehra and Gill 1968. Previous report of 2n= 24 confirm.
/Chamba: Manimahesh (2,600m) 54943 24 97.21 2x 24: Malik et al. 2011
48+2-4B: Dhillon 1979.
48: Bala and Gupta 2011.
S. koelzii Penn.
/Chamba: Pangi valley, Hillaur (2,650m) 54987 20 83.10 2x First report
S. polyantha Royle ex Benth.
/Parvati valley: Malana (3,300m) 54953 24+2-3B 74.20 2x 24+2B: Dhillon 1979. Previous report of 2n= 24 confirm.
*
PUN: Herbarium Code of Botany Department, Punjabi University, Patiala (Holmgren and Keuken 1974).

Table 2. Cytological information on the worldwide and India basis in Scrophularia L.

Genera in Species 2n counts Number of Species Base No/s.
Total Cytologically (distribution in species) 2x Polyloidy Aneuploidy
counted (%) Total ISP* Level
(%) (x)
** **
World 200 87 18 (2), 20 (1), 24 (13), 26 (23), 28 (1), 22 65 2(10), 2x, 12 7 , 8 , 9,
(43.50) 30 (2), 32 (1), 36 (3), 38 (1), 40 (8), (74.71) 2(13), 4x, 10, 12, 13,
42 (1), 43 (1), 44 (1), 46 (1), 48 (2), 1(15) 6x, 15, 19, 29
50 (1), 52 (3), 56 (3), 58 (27), 60 (3), 8x,
64 (1), 68 (1), 74 (1), 78 (1), 80 (2),
84 (3), 90 (2), 94 (1), 96 (1)
India 19 12 20 (1), 24 (4), 26 (3), 32 (1), 7 4 1(12) 2x, 2 8**, 10, 12, 13
(63.16) 40 (1), 48 (3), (33.33) 4x
* **
ISP= Intraspecific polyploids; Diploid (2n=2x) count is not yet reported. Common base numbers (X) are in bold while the most frequent one are underlined.

recorded with the inter-bivalent connections (Fig. 1d).
These connections were noticed to be present at the
terminal ends of bivalents. The phenomenon of cytomixis
was reported in 29.97% of the PMCs (Fig. 1e-g). Four to
five PMCs were involved in the phenomenon of cytomixis
at a time but transfer between two PMCs was most
common. No other meiotic anomaly was recorded in this
accession. In rest of the populations PMCs were observed
with normal meiosis and high pollen fertility (above 90%).
　 The chromosome counts of 2n=24 in the present
species were in conflict with the previous report of 2n=26
by Vasudevan (1975) from Kashmir Himalaya. However,
the chromosome numbers of 2n=24 and 26 are reported in
S. lucida L. which is some time confused with the
S. decomposita Royle ex Benth. (Shaw 1962; Varrama and
Hursalmi 1967; Grau 1982 and Snogerup 1955).
S. edgeworthii Benth.　Materials for meiotic study are
represented with two accessions, which were collected
from the two different populations growing in Malana
(2,650m) and Jhalma (2,700m) areas of Manikaran and
Udaipur, respectively. Both the populations were observed
with meiotic chromosome number of n=24 at diakinesis
(Fig. 1h-i). Nearly 14.03% PMCs in Jahalma population
were recorded with the phenomenon of inter-bivalent
connections at M-I (Fig. 1j-k). Meiotic course in other
population was meiotically normal. Both the populations
were recorded with high pollen fertility (above 95%).
　 The present chromosome count in both the accessions
are in agreement with the previous records of 2n=48 by
Mehra and Vasudevan (1972) from Kashmir, Himalaya.
 CHROMOSOME NUMBERS OF SOME SCROPHULARIA IN INDIA 43

S. himalensis Royle ex Benth.　The species is endemic Himalaya.

to Western Himalayas. Plant specimens were collected
from the population growing at Jahalma (2,650m) and
Manimahesh (2,600m) areas. The accession representing
these populations were observed with meiotic chromosome
number n=12 at diakinesis (Fig. 1l). Chromosome stickiness
was observed in a few PMCs (11.13%) at metaphase I in
the accession collected from Manimahesh area (Fig. 1m).
Meiotic course in both the population was normal and
high pollen fertility (above 100%) was recorded.
　 The present chromosome count in this species 2n=24
was in conformity with the recent report by Malik et al.
(2011) from Kashmir Himalaya. However, it is at variance
with earlier chromosome counts of 2n=20 (Mehra and
Gill 1968) and 2n=48+2-4B (Dhillon 1979) from western
S. koelzii Penn.　Accession of S. koelzii Penn. was
collected from the plants growing its populations at
Hillaur area (2,650m) of Pangi Valley. Meiotic preparations
in this accession revealed the presence of very small sized
chromosomes (2n=20) at metaphase I (Fig. 1n). The
accession was meiotically normal and about 83.10%
apparently fertile pollens were recorded.
　 The chromosome count 2n=20 for S. koelzii Penn. is
the first chromosome count for the species.
S. polyantha Royle ex Benth.　For the meiotic study the
plants were collected from Malana (3,300m) area of
Parvati Valley. Several PMCs revealed the presence of

Fig. 1. Meiotic chromosomes in pollen mother cells (PMC’s) in six species of Scrophularia in Lahaul-Spiti, H. P., India.
a. S. calycina: A PMC with 13:13 chromosome distribution at anaphase I. b-g. S. decomposita. b. A PMC showing 12
bivalent chromosomes at metaphase I. c. PMCs showing 12 bivalents at diakinesis. d. PMCs with inter-bivalent
connections at metaphase I. e. PMC’s involving in cytomixis at metaphase I. f-g. A group of 3 and more than 3 PMC’s
involving in cytomixis. h-k. S. edgeworthii. h-i. PMC’s with 24II at diakinesis. j-k. PMC’s showing inter-bivalent
connections at metaphase I. l-m. S. himalensis. l. A PMC showing 12II at diakinesis. m. PMC’s with chromosome
stickiness at metaphase I. n. S. koelzii : A PMC showing 20II at metaphase I. o-p. S. polyantha, PMCs with 12II +2B
chromosome at metaphase I.
44 SRIVASTAVA AND SAGGOO
12 bivalents (2n=24) at metaphase I and most of them
(17-32%) were recorded with 2B chromosomes at
metaphase I (Fig. 1o-p). Meiotically accession was normal
and showed 74.20% pollen fertility.
　 The chromosome number of 2n=24+2B in the present
species was in agreement with the previous reports of
2n=24+2B by Dhillon (1979) from Western Himalaya.
　 So far, 87 species (43.50%) of Scrophularia are known
for their chromosome counts and they exhibits a wide
range of chromosome numbers as 2n=18, 20, 24, 26, 28,
30, 32, 36, 38, 40, 42, 43, 44, 46, 48, 50, 52, 56, 58, 60,
64, 68, 78, 80, 84, 90, 94 and 96. The chromosome
numbers of 2n=24, 26 and 58 are frequent among the
above mentioned 2n counts. Three base numbers such as
x=9, 10 and 13 have been proposed by the various workers
(cf. Darlington and Wylie 1955; Larsen 1960; Löve and
Löve 1961). Carlbom (1968) gave the opinion of x=7 and
x=8 as the primary base numbers, but true diploids based
on these numbers are not available in the genus. Vaarama
and Hiirsalmi (1967) have added x=12 to this series as
another base numbers. The present data support the
polybasic nature with x=9, 10, 11, 12 and x=13. The
frequency of chromosome numbers based on x=13 and
x=29 is fairly higher than is x=12. Polyploidy in the genus
is mainly based on x=10 and x=13. Six species show
intra-specific polyploidy at different level of ploidy i.e. S.
grayana Maxim. ex Kom. (2n=20, 40; x=10), S. himalensis
Royle ex Benth. (2n=24, 48; x=12), S. maximowiczii
Gorschk. (2n=40, 80; x=10), S. scoparia Penn. (2n=60,
90; x=15), S. alata Gilib. and S. umbrosa L. (2n=26, 52;
x=13.). Aneuploidy is reported in 12 species. In India
nearly 12 species are cytologically available, apart from
the presently studies species other are S. dentata Royle
(2n=24), S. obtuse Edgew. (2n=48), S. scabiosaefolia
Benth. (2n=40), S. scopolii Hoppe (2n=26), S. variegata
Bieb. (2n=24) and S. variegata spp. depauperata (Boiss.)
Lall (2n=32). One species namely, S. himalensis is
reported with intra-specific polyploidy (2n=20, 24, 40,
48) coupled with aneuploidy while present report (2n=24)
in S. decomposita revealed its aneuploid nature (2n=26,
Vasudevan 1975).
　 From the above it is evident that, out of the six species
studied, one species is tetraploid (4x) based on x=12
(S. edgeworthii), while other five species are diploid (2x)
which are based on three different base numbers i.e. x=10
(S. koelzii Penn.), x=12 (S. decomposita Royle ex Benth.,
S. himalensis Royle ex Benth. and S. polyantha Royle ex Benth.)
and x=13 (S. calycina Benth.).
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