Trakia Journal of Sciences, Vol. 7 , No.

1 , pp 12-16, 2009
Copyright © 2009 Trakia University
Available online at:
http://www.uni-sz.bg
ISSN 1313-7050 (print)
ISSN 1313-3551 (online)
Original Contribution
POLLEN FERTILITY (VIABILITY) STATUS IN ASTERACEAE SPECIES
OF PAKISTAN
S. J. Qureshi1*, M. A. Khan2, M. Arshad3, A. Rashid4, M. Ahmad5
1
Pakistan Islamia Institute AlAin, Abu Dhabi, United Arab Emirates;
2
Department of Plant Sciences, Faculty of Biological Sciences, Quaid-i-Azam, University
Islamabad, Pakistan;
3
Department of Botany, PMAS Arid Agriculture University Rawalpindi, Pakistan;
4
Department of Environmental Sciences, PMAS Arid Agriculture University, Rawalpindi,
Pakistan;
5
Physics Division, PINSTECH, P.O. Nilore Islamabad, Pakistan
SUMMARY
Pakistan is facing widespread destruction and degradation of its flora. This study was conducted to
determine the pollen fertility status in forty five species of economic and environmentally important
species, which reproduced freely and thus most suitable for rehabilitation in Pakistan. Pollen fertility
status in these species varied as low as 42.3 % in Scorzonera ammophila to as high as 100 % in Crepis
flexuosa, C. multicaulis, Hieracium virosum, Lactuca altaica, L. lessertiana, L. orientalis, and L.
saligna. It was interesting to note that species within same genus showed wide range of pollen
viability. However, most of the species studied had high pollen viability especially in those species,
which grew in hilly areas. It was observed that pollen fertility was low in small fragmented
populations. This showed the effect of population size on reproduction. Larger size populations
showed high percentage of pollen fertility.

Key words: Pollen fertility, Viability, Asteraceae, Pakistan.

INTRODUCTION clearing of forests for plantations and

settlements has resulted in the loss of many
Pakistan is located in south Asia at the north
species.
of the Tropic of Cancer and lies between 230
The Asteraceae family consists of
to 370 north latitude and 610 to 760 east
approximately 25,000 species (3) included in
longitudes (1) It has an area of 796095 km2
over 1100 genera. These species frequently
1

covering about 0.6 % of the total area of the

present herbaceous habits, although
world. The flora is extremely diverse
arboreous and voluble herbaceous habits also
comprising more or less 6000 species (2)
occur (4). The Asteraceae (Compositae,
which include cultivated or naturalized taxa
alternate name) with its approximately 1,620
also and are found in a variety of habitats
genera and more than 23,600 species is the
from seashore and deserts to high
largest family of flowering plants (5). The
mountainous areas. About 372 species of
family is distributed worldwide except for
plants are endemic, mostly found in the
Antarctica but is especially diverse in the
northern and western mountainous regions of
tropical and subtropical regions of North
Pakistan. Various environmental factors of
America, the Andes, eastern Brazil, southern
Pakistan are being introduced here as every
Africa, the Mediterranean region, central
one can affect the distribution, growth,
Asia, and south-western China.
development and life cycles which lead to the
Pollen fertility is generally high in both
establishment of climax communities of
diploids and tetraploids. For example in the
plants. Pakistan also encounters widespread
diploid Tradescantia occidentalis, 94 % of
destruction and degradation of its flora. The
pollen was fertile, while 89 % pollen fertility
was observed in tetraploids (6). High fertility
* Correspondence to: Sohail Jamil Qureshi, of tetraploids pollen was also observed in
Pakistan Islamia Institute AlAin, Abu Dhabi,
Rhoeo discolor (Sw.) Stear, a close relative
United Arab Emirates, P.O.Box # 15778; Mobile:
00971506930626; E-Mail: of Tradescantia, where tetraploids produced
[email protected]
12 Trakia Journal of Sciences, Vol. 7, No. 1, 2009
 QURESHI S., et al.

twice as much fertile pollen as diploids (7). carmine staining technique in thirty two
Due to the effect of temperature on species of economic and environmental
chromosome pairing, pollen fertility in some importance. This investigation helps in
Tradescantia species may be higher in identifying coastal species, which are
periods of warm weather than after a series of reproducing freely and are therefore most
cold, wet days (6). suitable for coastal reforestation and
The frequent removal of species rehabilitation.
together with the lack of understanding of
consequences and advantages of replanting MATERIALS AND METHODS
led to serious coastal degradation and to the
Different species of six genera belonging to
endangerment of a high proportion of species.
Asteraceae (which were in flower during the
Therefore there is urgent need for
time of this study) were tested for pollen
conservation measures to rehabilitate the land
fertility status by using the aceto-carmine
and restore plant communities. Besides
staining method. The aceto-carmine method
human activity, other factors have also
of determining viability in different species
contributed to the degradation of Pakistan
of plants is comparable to other methods such
flora. Periodic natural disasters such as
as pollen germination method on artificial
hurricanes and storms, although infrequent,
medium (18) and Flow Cytometry Reactivity
caused enormous devastation and destruction
(19).
to plants and animals (8-11). In October,
Flower buds, flowers or inflorescences
1972 Hurricane Bebe swept across the
of these species were collected from Pakistan
Rotuma Island, its strong winds and
and where possible their pollen was analysed
accompanying unusually high waves caused
for viability within one of collection or kept
extensive damage to homes and vegetation
in the fridge at 4o C for analysis the next day.
(12).
The method followed that closely used by
Pollen, which is a carrier of male
Marutani (16). Using dissecting forceps,
gametes, includes three domains that are
scalpel and a needle, anthers of various
different in their chemical composition,
species were opened to allow extraction and
morphological structure and their
subsequent transfer of pollen dust on to a
physiological and biological significance
microscopic glass slide in a drop of aceto-
(13). The three domains of pollen grain
carmine stain. Mature anthers were crushed
include exine, intine and nucleus. The
and pollen grains mixed thoroughly with the
complex exine structures of pollen are
aceto-carmine stain. Cover slips were gently
storage sites for carbohydrates, glyco-
placed on to different slides for each species.
proteins, lipids, terpenoids and phenolics
The slides were than observed under a light
(14). The pollen nucleus is rich in chromatin
microscope. For each plant species flowers
material and viable pollen stains pink to deep
were collected from at least three different
red with aceto-carmine, while srerile (mostly
plants. For each plant, three slides were
shrivelled) pollen does not take any stain and
prepared. For each slide ten randomly
thus remains almost white and transparent
selected fields were observed under the 10 X
(15, 16). A viable or fertile pollen is one
objective (100 X magnification).
which, after landing on stigma of the same
To determine pollen fertility, darkly
plant or other plants of the same variety or
stained pollen grains were recorded as fertile
species, under normal conditions would start
and viable, and unstained or very lightly
growing a pollen tube and finally discharge
stained ones were considered as sterile or
its male gametes in the embryo sac effecting
non-viable. Pollen fertility was calculated by
fertilization.
dividing the number of viable pollen grains
According to Rigamoto and Tyagi,
by the total number of grains counted in the
(17) pollen fertility, which can be determined
field of view and averaging them for all
using pollen viability tests in-vitro is very
plants in that species. Pollen viability was
important in fruit and seed production in
expressed as percentage pollen fertility in
flowering plants. Therefore, pollen fertility
each plant species. Standard error was
knowledge for any plant species is essential
calculated from original data to compare for
for plant breeders and commercial growers.
significant differences among different plant
They determine pollen fertility status using
species. Collections were made from
pollen viability tests in-vitro by aceto-
different sites, given in the map (Figure A

Trakia Journal of Sciences, Vol. 7, No. 1, 2009 13

 QURESHI S., et al.
RESULTS DISCUSSION
Pollen fertility results obtained using the
On Table 1, the pollen fertility (viability)
aceto-carmine method are presented on Table
status of most species tested was high. Several
1. Average pollen viability ranged from a low
factors contributed to the low number of
42.3+2.3 for Scorzonera ammophila to 100 %
species included in this pollen fertility count.
for Crepis flexuosa, C. multicaulis, Hieracium
Theoretically it was possible to analyse all
virosum, Lactuca altaica, L. lessertiana, L.
plant species. However, due to several factors
orientalis and L. saligna. Out of the 45
it was not possible during the span of this
species analysed for their pollen viability,
study. Firstly, even though some species were
only one species (2.2 %), namely Scorzonera
in flower, they were usually too tall for easy
ammophila showed a pollen viability
flower collection. Secondly, most of the
percentage lower than 50 %. Fifteen species
species were in the fruiting season at the time
(33.3 %) showed pollen viability of 50-80 %.
of this survey. Thirdly, the flowers of species
The remaining twenty nine species (64.4 %)
were typically small having microscopic
showed a pollen viability percentage of 80-
anthers. This made the extraction, crushing
100 %.
and counting of viable pollen grains a slow
and tedious process. However, most of the
species were analysed for their pollen
viability.
In view of the impoverished flora of Pakistan,
and the urgent need to reforest and replenish
it, knowledge of the pollen fertility status of
Pakistan plant species would be very useful.
Pollen fertility is a significant determinant of
whether in a population there will be enough
regeneration through sexual reproduction to
ensure the survival of that species. Pollen
fertility is also evidently reduced in small
fragmented populations (20). Other workers
have investigated the effect of population size
on reproduction of some angiosperms and
found that reproduction was strongly reduced
Figure A: Location of sampling sites from in small population where plants produced
Pakistan. fewer seeds per fruit per plant (21-23). They
Species within a single genus showed concluded that insufficient quantity and
different pollen viability. For example in the quality of pollen was the likely explanation
Lactuca genus, L. altaica has an average for the reduced fecundity in small
pollen viability of 100 %, + 0.0 with a range populations. Fischer and Mathies (24)
of 100-100 % while the least average value of observed that fecundity was four times higher
pollen viability in L. dissecta is 52.6 %. The in large populations of the rare Gentianella
seven species of different genera, namely germanica than in small populations and that
Crepis flexuosa, C. multicaulis, Hieracium fitness was reduced in plants from small
virosum, Lactuca altaica, L. lessertiana, L. populations. Menges (25) and Agren (26) also
orientalis, and L. saligna showed 100 %, +0.0 observed low pollen fertility and reduced seed
pollen viability. However, a larger difference germination rates in small fragmented
was observed in different species of Lactuca populations.
genus. On the other hand, the seven species in Seed formation or regeneration
pairs have same average pollen viability i.e. information is not available; thus the main
69.3 % (Lactuca serriola L. f. integrifolia and emphasis of this study was the concern that
Sonchus arvensis), 72 % (Hieracium plants with low pollen fertility might become
umbellatum and Lactuca brevirostris), 78 % scarcer and could become endangered. Pollen
(Scorzonera laciniata and Sonchus asper), fertility status of the plant species from
78.3 % (Scorzonera virgata and Sonchus Pakistan is required further investigation to
palustris), 80.3 % (Sonchus oleraceus and include all the species. This will help to
Taraxacum aff var. glabrescence), 93.3 % identify all the species with low pollen
(Taraxacum eriopodum and Taraxacum aff. fertility, which would need replenishing.
var. typica), 94.6 % (Hieracium vulgatum and
Tragopogon dubius).
14 Trakia Journal of Sciences, Vol. 7, No. 1, 2009
 QURESHI S., et al.

Table 1: Pollen fertility (viability) percentage in 45 Asteraceae plant species of Pakistan

Scientific Names Pollen Fertility Percentage
Range Mean + S.E.
Crepis flexuosa (Ledebour) C. B. Clarke 100-100 100+0.0
C. multicaulis Ledebour 100-100 100+0.0
C. sancta (L.) Babcock 74-80 77.3+2.0
C. stocksiana Aitch. & Hemsl 58-67 63.3+2.5
C. thomsoni Babcock 76-88 82.6+3.5
Hieracium bichlorophyllum (Druce & Zahn) 63-79 73+4.1
Pugsley
H. diaphanoides Lindeb 90-95 92.6+2.3
H. umbellatum L. 68-75 72+1.7
H. virosum Pallas 100-100 100+0.0
H. vulgatum Fries 91-97 94.6+2.5
Lactuca altaica Fischer & C. A. Meyer 100-100 100+0.0
L. brevirostris Champ 67-75 72+2.0
L. clarkei Hooker 53-61 57.3+2.2
L. dissecta D. Don 48-56 52.6+2.4
L. dolichophylla Kitam. 47-59 54.3+3.2
L. graciliflora DC. 79-84 81+1.2
L. lessertiana C.B. Clarke 100-100 100+0.0
L. orientalis Boiss 100-100 100+0.0
L. remotiflora DC. 89-97 93.6+2.8
L. saligna L. 100-100 100+0.0
L. sativa L. 68-72 70+1.0
L. serriola L. 62-74 69+2.9
L. serriola L. f. integrifolia (Gray) S. D. Prince & 66-72 69.3+1.9
R. N. Carter
L. tatarica C.A. Mey 77-86 82+2.1
L. undulata Ledeb 61-68 65+1.7
L. virosa L. 72-84 79.3+4.0
Scorzonera ammophila Bunge 38-47 42.3+2.3
S. hondae Kitam 53-61 57.6+2.5
S. laciniata L. 71-83 78+3.0
S. picridioides Boiss 78-89 84+2.6
S. virgata DC. 75-81 78.3+2.0
Sonchus arvensis L. 66-72 69.3+2.0
S. asper (L.) Hill 74-81 78+1.7
S. maritimus L. 88-95 92+1.7
S. oleraceus L. 75-84 80.3+2.5
S. palustris L. 76-80 78.3+1.6
S. uliginosus M. Bieb. 58-65 62+1.7
Taraxacum eriopodum (D. Don) DC. 87-99 93.3+3.2
T. aff var. glabrescence Web. 77-84 80.3+2.1
T. glaucescens Wigg 68-87 76.3+4.7
T. laevigatum (Willd.) DC 74-79 76.6+2.2
T. aff. var. parvula Wigg and Boiss 81-97 90+3.8
T. aff. var. typica Hk 89-97 93.3+2.4
Tragopogon dubius Scop 88-99 94.6+3.4
T. gracilis D. Don 76-90 83.6+3.8

ACKNOWLEDGEMENT
We are grateful to Chairman, Department of and palynological lab. He also made
Plant Sciences, Faculty of Biological constructive comments on an early version of
Sciences Quaid-i-Azam University the manuscript.
Islamabad, for his assistance in the herbarium

Trakia Journal of Sciences, Vol. 7, No. 1, 2009 15

 QURESHI S., et al.
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