8
The 7th Inter. Conf.”Plant & Microbial Biotech. & their Role in the Development of the Society”pp.103 -110 (2017)
Cytological Studies on Some Egyptian species of Anthemideae
(Asteraceae)
A.M. Atta, M.H. Abd El-Twab*, R.K. Helmey*# and H. Dahy*
Genetics Department, Faculty of Agriculture and *Botany & Microbiology
Department, Faculty of Science, Minia University, Minia, Egypt.
T
HE MEIOTIC behavior and morphology of pollen grains of four species of Asteraceae
(Glebionis coronaria, Anacyclus monanthos, Matricaria recutita and cotula barbata)
were studied. Glebionis coronaria, Anacyclus monanthos and Matricaria recutita showed nine
chromosome bivalents while those of Cotula barbata showed eight chromosome bivalents.
The chromosomes were meiotically identiied as two pairs in three taxa (Glebionis coronaria,
Anacyclus monanthos and Matricaria recutita) while it was one pair in Cotulabarbata.
The pollen shape and morphology were recognized by measuring polar and equatorial axis,
Morphological characteristics of the pollen grains of the four studied species were considerably
different.
Keywords: Cytological studies, Egyptian species of Anthemideae, Meiotic, Pollen morphology.
Introduction
Chromosome numbers and karyotype analysis of
somatic chromosomes have been described for
several species of Asteraceae in Egypt. Detailed
karyotype features of 23 species of subfamily
Asteroideae were described by Badr et al. (1997)
and for 24 species from the tribes Chicorieae
(Lactuceae) and Cardueae (Cynareae) by Badr
et al. (2009). Comparative karyological analysis
was also described for selected species such as
Artemisia judaica and Artemisia monosperma
(Badr et al., 2012). Additional counts were added
on Cotula barbata, Achillea fragrantissima,
Achillea santolina, Anacyclus monanthos,
Artemisia arborescens, Glebionis coronaria,
Cotula cinerea and Matricaria recutita (Abd El
Tawab et al., 2008; 2012 and Badr et al., 2014).
Diploid and polyploid somatic chromosome
numbers based on x=5, 7. 8, 9 and 11 have been
reported and karyotype symmetry indicators
i.e. the values of the mean arm ratio, TF%, A1
and A2 indices, clearly demonstrated that the
karyotype is generally symmetric with small
variations between the chromosomes in length
and centromere position (Badr et al., 1997 and
2009). However, none of the above-mentioned
studies have addressed meiotic chromosomes.
According to the observations of numerous
authors, palynological studies provide an
excellent set of characters that complement
traditional morphological studies, and contribute
to the understanding of the taxonomy, phylogeny
and ecology of the Asteraceae. The complexity
of the Asteraceae pollen often includes the
characteristics of the aperture system, an issue
widely discussed by several authors because
of the different techniques of observation used
and the heterogeneity of this taxonomic group
(Montes & Murray, 2105).
The main goal of this work is to investigate
the behavior of meiotic chromosomes and pollen
grain morphology of four species of Asteracaea
collected from different localities of Egypt.
Materials and Methods
Plant material
Plants of four species from the tribe
Anthemideae, family Asteraceae were collected at
lowering and fruiting stages from plants growing
in their natural habitats and identiied using the
manual Students Flora of Egypt (Täckholm,
1974). Plant species used in this cytological study
are Glebionis cronaria (L.), Matricaria recutita
(L.), Anacyclus monanthos (L.) Thell., and Cotula
barbata DC.(Table 1 and Fig. 1).
#
Corssponding auther email: rashahelmey@yahoo.com
DOI: 10.21608/ejbo.2017.837.1058
©2017 National Information and Documentation Centre (NIDOC)
104
A.M. ATTA et al.
TABLE 1. Plants from different taxa at tribe Anthemideae from family Asteraceae and their localities in Egypt
Species
Location
Glebionis coronaria
Minia University and Cairo-Alexandria desert Road
Matricaria recutita
Minia University
Anacyclus monanthos
Borg El-Arab
Cotula barbata
Minia University-Botanical garden of Agriculture faculty
A
C
B
D
Fig. 1. Photographs of plants of the studied taxa. A. Glebionis coronaria, B. Matricaria recutita, C. Anacyclus
monanthos and D. Cotula barbata. Bar equals 4 microns.
Egypt. J. Bot. (2017)
CYTOLOGICAL STUDIES ON SOME EGYPTIAN SPECIES ...
105
(there are two chiasmata or more located along
with chromosome or rod (pairing between the two
homologous chromosomes is at one chromosome
end and only one terminal chiasma is observed).
PMCs of species of G. coronaria showed nine
bivalents at diakinesis (Fig.2A). Mean number of
rod and ring bivalents at diakinesis in species of
G. coronaria were 2.795 and 6.205, respectively.
The chiasma frequency per cell and per bivalent
were 15.26 and 1.695, respectively as shown in
Table 2. PMCs of species M. recutita showed nine
bivalents at diakinesis (Fig. 2B).Mean number of
rod and ring bivalents in species M. recutita were
3.3 and 5.7, respectively. The chiasma frequency
per cell and per bivalent were 14.69 and 1.63
respectively (Table 2). Similarly, PMCs of plants
of species A. monanthos showed nine bivalents
and the nucleolus is associated with two bivalents
at diakinesis (Fig. 2C). Mean number of rod
and ring bivalents in A. monanthos were 3.231
and 5.769, respectively. The chiasma frequency
per cell and per bivalent were 14.77 and 1.641,
respectively (Table 2). PMCs of plants of species
C. barbata showed eight bivalents at diakinesis
(Fig. 2D). The nucleolus was seen associated
with one bivalent. Mean number of rod and ring
bivalents in species C. barbata were 7.231 and
0.7692, respectivelly and the chiasma frequency
per cell and per bivalent were 10.85 and 1.356,
respectively (Table 2).
Meiotic preparation
Young lowering buds (45 days aged plants)
were collected and immediately ixed (3 ethyl
alcohol: 1 acetic acid) for 24 h at 4ºC. The
meiotic behavior of chromosomes in pollen
mother cells (chromosome pairing) was examined
in at least (50 cells / species). Aceto-carmine
stained microscopic slides were prepared from
mature anthers and viable pollen grains (with
the stained protoplast) were counted. The
number of bivalents, the ring and rod units, and
chiasma frequency per cell and per bivalents
were recorded. Some PMCs were photographed
using Olympus C-4040 Camera. Pollen grain
morphology was also examined in large lowering
buds and photographed. Measurements of pollen
grains were taken using SIS image analysis.
Chiasma frequency per bivalent, ring frequency,
rod frequency and LSD were calculated using
MSTAT program.
Results and Discussion
Pollen mother cells (PMCs) of plants of
species G. coronaria, A. monanthos and M.
recutita showed nine bivalents while those of
species C. barbata showed eight bivalents at
diakinesis and metaphase I (Fig.2). According to
chiasma number, position and pairing pattern, the
chromosome bivalents were classiied either ring
A
B
C
D
Fig. 2. Stages of meiosis (Diakinesis) showing ring and rod bivalents. A. Glebionis coronaria., B. Matricaria recutita, C. Anacyclus
monanthos and D. Cotula barbata. Bar equals 4 microns.
Egypt. J. Bot. (2017)
106
A.M. ATTA et al.
TABLE 2. Mean number of ring and rod bivalents and chiasma frequencies of the four species of Asteraceae.
Species
Glebionis
coronaria
Matricaria
recutita
Anacyclus
monanthos
Cotula barbata
LSD 0.05
Number of
counted cells
Chiasma
frequency per
cell
Chiasma
frequency per
bivalent
Ring
Rod
50
15.26
1.695
6.205
2.795
50
14.69
1.632
5.692
3.308
50
14.77
1.641
5.769
3.231
50
10.85
2.98
1.356
0.7692
0.43
7.231
0.42
The pollen grains of four representative species
belonging to Asteraceae were shown in Fig.3. The
pollen shape and morphology were recognized by
measuring polar and equatorial axis (Table 3). All
pollen grains of Glebionis are oblate-spheroidal
and the pollen walls are provided with spines
(Fig. 3A.). The polar axes 22.23- 29.95µm and the
equatorial axes 24.65- 31.79 µm.The pollen grains
of M. recutita are operculate and tricolporate and
the pollen wall is provided with spines with the
polar axes 15.12- 20.88 µm and the equatorial
axes 16.18- 22.76 µm (Fig. 3B). All pollen grains
of Anacyclus are oblate-spheroidal with the polar
axes 11.8- 14.88 µm and the equatorial axes 12.515.31 µm. The pollen wall is smooth (Fig. 3C).
All pollen grains of C. babata are sub-oblate with
the polar axes 15.9- 23.03 µm and the equatorial
axes 18.53- 25.4 µm, they are operculate and
tricolporate and the pollen wall is provided with
spines (Fig. 3D).
A
B
C
D
Fig. 3. Pollen microphotography of A. Glebionis coronaria., B. Matricaria recutita, C. Anacyclus monanthos and D. Cotula barbata.
Bar equals 4 microns.
Egypt. J. Bot. (2017)
Echinate
Tricolporate
19.47
23.03
18.53
25.4
21.97
0.88
Suboblate
Smooth
Tricolporate
Oblatespheroidal
13.34
14.88
12.5
15.31
13.91
0.92
Echinate
Tricolporate
Oblatespheroidal
18
11.8
15.9
18
18
16
Matricaria
recutita
Anacyclus
monanthos
Cotula barbata
15.12
20.88
16.18
22.76
19.47
0.92
Echinate
Tricolporate
Oblatespheroidal
26.09
18
Glebionis
coronaria
22.23
29.95
24.65
31.79
28.22
0.92
Ornamentation
Mean
Max
Min
Mean
Max
Min
Equatorial axes (E) (µm)
Polar axes (P) (µm)
Chromosome
No. (2n)
Taxa
TABLE 3. Pollen measurements in (µm) of four species belonging to Asteraceae.
P/E
Pollen
shape
Aperture
type
CYTOLOGICAL STUDIES ON SOME EGYPTIAN SPECIES ...
107
Morphological characteristics of the pollen
grains of the four studied species were considerably
different. These data are in accordance with those
reported by Bose et al.(2012). These differences
are used to differentiate between genera and even
species. For example, in Matercaria species,
seven major pollen types have been recognized
on the basis of type and number of aperture shape,
aperture type and ornamentation (Ceter et al.,
2013).
Cytological data presented herein are in agreement
with several works (Abd El-Twab et al, 2008;
Humphries, 1980; Powell et al, 1974 and Schweizer
& Ehrendorfer, 1976). The observed regular meiosis
suggested that intra- and inter-genomic reorganization
process occur quickly and eficiently in these
taxa. The higher number of the ring bivalents at
diakinesis and metaphase I may be attributed to
median centomeric positions on the chromosomes
of these taxa (López et al., 2013). As a rule,
metacentric and submetacentric chromosomes
formed ring bivalents, while acrocentrics and
subtelocentrics paired in rod units during irst
meiotic prophase and metaphase I. The karyotype
structures, composition and distribution of the
heterochromatin in Glebionis coronaria and
Glebionis segetum using Giemsa and luorescent
banding techniques were also analyzed by Paciolla
et al. (2010). They concluded that G. coronaria
possesses the most symmetrical karyotype,
comprising mainly metacentric chromosomes.
The formation of rod bivalents in spite of rings
in the existence of metacentric and submetcentric
chromosomes observed at diakinesis and
metaphase I in the present materials may due to
different pairing conditions, degree of homology
and recombination systems.
Chiasma frequency per chromosome depends
on the length and homology of chromatin content.
The data revealed that the lowest value of chiasma
frequency per chromosomes was in plants of C.
barbata. This is not due to low haploid number
(n=8) but to chromosome length. The frequency
of chiasma differences were extensively analyzed
in several taxa belonging to Asteraceae (Ghaffari
& Djavadp, 1998; Gupta et al., 2013; Kumari
& Sidhu, 2012 and Sheidai et al, 2006). They
concluded that the cytotaxonimic features verify
the afinity of the related taxa of Asteraceae. The
principal taxonomic problems within the tribe
Anthemideae are almost entirely relationships
between genera, but also circumscription of
Egypt. J. Bot. (2017)
108
A.M. ATTA et al.
genera, especially within subtribal groups such
as the Artemiisinae, Chrysantheminae and
Tanacetinae (Bremer & Humphries, 1993).
Although the Anthemideae tribe has been reviewed
extensively (Bremer & Humphries, 1993), small
karyotype changes appear already within species
and gradually reduce chromosome pairing afinities
in species hybrids. Gross translocations and loss
of centromeres have led to species divergence
and descending dis-ploidy (2 n = 16 ~ t2). Some
species of Anacyclus were karyologically examined
by Humphries (1980) and indicated that evolution
in genus Anacyclus has been accompanied by
chromosomal re-patterning. The understanding
of the relationships of the annual of these taxa is
complicated by sympatry and the occurrence of
natural hybrids.
Chromosomes with nucleolar organizer were
meiotically identiied as two pairs in the three taxa
G. coronaria, A. monanthos and M. recutita while it
was one pair in C. barbata. Chromosome numbers,
morphology and nucleolus organizer region (NOR)
locations are useful cytological characters for
taxonomic and evolutionary studies (Barth et al.,
2013). Recently, molecular cytogenetic localization
of rDNA genes was achieved in different species
belonging to Asteraceae (Bernardes et al., 2013
and Abd El-Twab et al., 2015). Thus, the presence
of repetitive DNA clusters (rDNA sites and
luorochrome bands) suggested the great importance
of such DNA elements during the karyotype
evolution. The karyological markers developed
allowed karyotypic differentiation between both
species, uncovering peculiarities in the number and
position of rDNA sites, which suggests chromosome
rearrangements, such as translocations (Bernardes
et al., 2013). Genetic variation within and between
plant populations is essential for establishment the
genetic diversity database of plant genetic resources
(Badr et al., 2012; 2014 and 2016).
Conclusion
Cytological coniguration at irst meiotic
division and pollen morphology were described in
four species of Asteraceae (Glebionis coronaria,
Anacyclus monanthos, Matricaria recutita and
cotula barbata). The number, type, position and
distribution of chismata in the bivalent were
described. In addition the shape and characteristics
of pollen in the examined species are described in
detail.
Egypt. J. Bot. (2017)
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دراسات خلوية على بعض اأنواع المصرية من نباتات العائلة المركبة
عبد التواب محمد عطا ،مجدي حسين عبد التواب* ،رشا كمال حلمي* و هدير ضاحي
قسم الوراثة -كلية الزراعة و*قسم النبات و الميكروبيولوجي – كلية العلوم – جامعة المنيا -المنيا -مصر.
*
تم دراسة كل من سلوك الكروموسومات أثناء اانقسام الميوزي و الشكل الظاهري لحبوب اللقاح وذلك في أربعة انواع من
نباتات العائلة المركبة وهي (جليبيونس كوروناريا ،أناسيكلس مونانثوس ،ماتريكاريا ركيوتيتا و كوتيوا بارباتا ) .و لقد
أشارت الدراسات السيتولوجية إلى ظهور 9وحدات ثنائية كروموسومية فى كل الخايا اأمية لحبوب اللقاح فى نباتات اأنواع
)جليبيونس كوروناريا ،أناسيكلس مونانثوس و ماتريكاريا ركيوتيتا) ،بينما تم رصد 8وحدات ثنائية فقط فى الميوزى اأول فى
نباتات كوتيوا بارباتا.
ولقد أظهرت حبوب اللقاح تباين فى كثافة الكروماتين والشكل الخارجى فى النباتات تحت الدراسة وذلك عند تعيين قياسات
لكل من المحور القطبى و ااستوائى لحبوب اللقاح.
)Egypt. J. Bot. (2017
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