Egypt. J. Bot. Vol. 59, No.2, pp. 475 - 482 (2019)
35
Dicliptera aegyptiaca (Acanthaceae), A New Species from Egypt
Supported by Morphological Characters and rbcl-based DNA
Barcoding
Eman M. Shamso(1), Ahmed S. Fouad(2)#
The Herbarium, Botany and Microbiology Department, Faculty of Science, Cairo
University, Giza 12613, Egypt; (2)Botany and Microbiology Department, Faculty of
Science, Cairo University, Giza 12613, Egypt.
(1)
D
ICLIPTERA aegyptiaca, a new species from Red Sea Coast, Egypt, is described and
illustrated. Diagnostic and morphological characters that distinguish it from its allied
species D. paniculata and an identification key for the two species are provided. The new species
differs from D. paniculata by having an unbranched stem, a congested inflorescence with dwarf
axes 1.5– 5mm long; subsessile cymules with peduncles 0.5– 1mm long. rbcl-DNA barcoding
is presented for this new taxon for the first time. Phylogentic tree revealed barcode clusters for
the two Dicliptera species and recognized significant interspecific variation between them. D.
aegyptiaca clearly formed one clade strongly supported with a bootstrap value of 100%. Based
on characters of morphology, pollen and seeds, the new species was recognized as belonging to
the genus Dicliptera. On the other hand, DNA barcoding reflected clustering of all Dicliptera
spp. in a large clade while D. aegyptiaca formed a non sister clade showing the utility of DNA
barcoding for species identification rather than taxonomy.
Keywords: Dicliptera aegyptiaca, DNA barcoding, Egypt, Morphology, New species.
Introduction
Peristrophe are together monophyletic.
The genus Dicliptera Juss. comprises about 175
species (Daniel, 2009), distributed in tropical
and subtropical countries of Asia, Africa and
America. Within Acanthaceae, it is placed in the
Diclipterinae clade of the “Justicioid” lineage with
Hypoestes as a sister to Peristrophe (McDade et
al., 2000 a). In the absence of fruits, it is difficult
to distinguish between Peristrophe and Dicliptera
s. str. (Balkwill, 1996 and Darbyshire & Vollesen,
2007). Based on the mechanism of capsule
dehiscence, Balkwill (1996) and Balkwill et al.
(1985, 1986, 1996) maintained that the two taxa
should remain separate, where the capsule is elastic
in Dicliptera and inelastic in Peristrophe. In their
work, Darbyshire & Vollesen (2007) transferred all
recognized Peristrophe from Tropical East Africa
to Dicliptera citing inconsistencies of capsule types
and the presence of intermediate forms. The most
recent molecular data on the “Justicioid” lineage,
Kiel et al. (2017) support Darbyshire & Vollesen’s
treatment as sampled species of Dicliptera and
Dicliptera (inclusive of Peristrophe) is
characterized by angled stem, opposite leaves,
inflorescence panicle-like cymose, with 2–3
inflorescence units (cymules), umbellately
arranged, and a resupinate bilabiate corolla. Fruitcapsule clavate, 4-seeded.
#
In Egypt, the genus Dicliptera was represented
by only one species D. paniculata (Peristrophe
paniculata), a rare species known only from Gebel
Elba and Red Sea Coast of Egypt (Täckholm,
1974; El Hadidi & Fayed, 1994/ 95; Boulos, 2002,
2009; Shamso, 2010, 2013 and El-Gazzar et al.,
2015). As part of the revisionary study of the genus
Dicliptera in Egypt, Shamso (2010) recorded
Dicliptera paniculata with a wide range of variation
in many morphological characters. Most examined
specimens represented D. paniculata, while some
other specimens did not match and were probably
an undescribed taxon. Their identities were
confirmed by referring to various regional floras
Corresponding author email: ahmedsfouad@yahoo.com
DOI: 10.21608/ejbo.2019.5371.1220
Edited by: Prof. Dr. Adel El-Gazzar, Faculty of Science, El-Arish University, El-Arish, N. Sinai, Egypt.
©2019 National Information and Documentation Center (NIDOC)
476
EMAN M. SHAMSO AND AHMED S. FOUAD
(Clarke, 1901; Andrews, 1956; Hein, 1966; Abedin
et al., 2000; Hedrén, 2006; Ensermu Kelbessa,
2006 and Darbyshire et al., 2010, 2015) and much
other relevant literatures (Balkwill, 1996; Balkwill
& Getliffe Norris, 1989; Balkwill et al., 1985, 1986,
1988, 1996; Ensermu Kelbessa, 2003; Darbyshire
& Vollesen, 2007 and Al Hakimi et al., 2017). The
specimens were also compared with herbarium
specimens and images available in virtual herbaria
[Kew Herbarium Catalogue (http://apps.kew.org/
herbcat/navigator.do), African Plant Database
(www.tropicos.org) and JSTOR Global Plants
(https://plants.jstor.org/)]. It is suggested that these
specimens do not match with any of the described
species of Dicliptera (incl. Peristrophe) and so are
described here as a new species.
DNA barcoding provides an accurate, rapid
and time-efficient tool for species identification
utilizing short DNA sequences as internal species
tags (Hebert, 2003 and Hebert & Gregory,
2005). The CBOL plant working group (2009)
recommended employing rbcl and/or matk in
barcoding of land plants. Compared to matk, rbcl
has high amplification success rate; it is the most
characterized plastid coding region in GenBank
(Newmaster et al., 2006 and Kang et al., 2017). rbcl
was used for identification of cryptic species by
several research groups (Miwa et al., 2003; Costea
& Stefanovic, 2009; Liu et al., 2013 and Ardiyani
et al., 2017).
The aims of this study are to confirm the
occurrence of the new taxon Dicliptera aegyptiaca
using macro- and micro-morphological data
and DNA barcodes, to provide the validating
description of the new taxon and to provide a
key to the Egyptian species of Dicliptera for easy
recognition of the taxa in Egypt.
Materials and Methods
Plant materials
The present study was based on examination
of several specimens belonging to Dicliptera
paniculata kept in the major Egyptian herbaria
[Cairo University Herbarium (CAI), the Agricultural
Research Centre, Flora and Phytotaxonomy
Herbarium (CAIM), National Research Centre,
Plant Systematic Herbarium (CAIRC) and Sohag
University Herbarium (SHG)]. The new taxon is
accompanied by English description (ICBN, 2018:
article 39.2).
Egypt. J. Bot. 59 , No.2 (2019)
Pollen and seeds examination
Samples of pollen and seeds were taken from
mature anthers and capsules, respectively. For light
microscopy (LM), anthers were boiled for a few
minutes in water, macerated in a few drops of an
aqueous 10% solution of KOH on a clean slide,
then stained with Safranin (1% Safranin solution
in 50% ethanol), mounted in glycerin jelly and
observations were made with a Sterico research
microscope under (E 40, 0.65) using a 16x eye
piece, and the seeds were examined with the aid
of a dissecting microscope. For SEM studies, both
pollen and seeds were mounted onto stubs with
double sided adhesive, and then these stubs were
sputter-coated with gold (Ion-sputter JFC-1100).
After coating, they were examined using JEOL
JSM 5400LV scanning electron microscope at
15KV, at the Electron Microscopy Unit, Assiut
University. The description of pollen follows the
terminology of Hesse et al. (2009) and seed coat
terminology follows Barthlott (1981).
DNA barcoding
Each of Dicliptera paniculata and the new
species Dicliptera aegyptiaca were represented by
two herbarium specimens. From each specimen,
about 20mg were collected and ground under
liquid nitrogen using a mortar and pestle until a
fine powder was produced. DNA was extracted
using a Qiagen DNeasy kit (Valencia, California,
USA) as outlined by manufacturer’s protocol with
few modifications for herbarium tissues. To the
AP1 buffer DDT (Melford Laboratories, UK) at
0.12mg/ml and Proteinase K(Sigma) at 0.04mg/
ml were added and shaken with ground tissue
for 60min at 65°C (de Vere et al., 2012). rbcl
amplifications were performed following CBOL
Plant Working Group (2009) with the specific
primers:
5’-ATGTCACCACAAACAGAAAC3’and 5’-TCGCATGTACCTGCAGTAGC-3’ in a
reaction mixture containing 25 µl PCR Master Mix
(Bioline), 1µl of each primer and 20-50ng genomic
DNA; the volume was completed to 50µl with
sterile distilled water. The amplification protocol
was 95°C for 2min followed by 34 cycles of 94°C
for 1min, 55°C for 30sec and 72°C for 1min, then
final extension for 7min at 72°C. Amplification
products were purified using the QIAquick PCR
Purification Kit (Qiagen, Hilden, Germany) then
sequenced with Big-dye terminator chemistry
in 3130xl Genetic Analyzer (Life Technologies,
California, USA) by following the standard
manufacturer’s protocol. Forward and reverse
sequences were assembled using Codon Code
DICLIPTERA AEGYPTIACA (ACANTHACEAE), A NEW SPECIES...
Aligner software, v. 7.1.2., the contig sequences
were deposited in the GenBank database under
accession numbers MH028051 and MG1990431.1
for Dicliptera paniculata and KU947958.1
and KU947961.1 for Dicliptera aegyptiaca.
The available online sequences for Hypoestes,
Peristrophe and Dicliptera in Gene bank were
employed to construct a phylogenetic tree and
calculate pairwise distances using Maximum
Likelihood (ML) method and Cluster W in MEGA
v. 6 (Tamura et al., 2013) based on Tamura 3
parameter model (Tamura, 1992) with gamma
distribution. Significance was assessed using 1000
bootstrap replications (Felsenstein, 1985).
Results and Discussion
Dicliptera aegyptiaca E. Shamso sp. nov.
Type: Egypt: Wadi El Faraied, Red Sea coast
23o31’0’’N; 35o19’60’’E; 12/2/1961; Täckholm et
al. 856 (holotype CAI).
Annual herb, 20– 30cm tall, unbranched,
rarely with short lateral branches above. Stem
angled, glabrous, sparsely strigose with appressed
eglandular multicellular hairs on angles and
nodes, cystolith present. Leaves rapidly deciduous
at lower nodes, blackish-green when dry. Leaf
lamina narrowly ovate to lanceolate 15– 45x 9–
15mm, attenuate at base, acuminate apex, minutely
glandular pubescent, strigose with eglandular
multicellular hairs at nerves and margins. Petiole
2– 6mm long, strigose (Fig. 1 A). Inflorescence
congested in the axils with dwarf axes 1.5–
5mm long, inflorescence of (1 –) 2 – 3 cymules
(inflorescence units), umbellately arranged,
often compound, with many umbels in each axil.
Inflorescence bracts (secondary bracts) 2, subequal,
subulate, 3– 5x 0.3– 0.5mm, sparsely hairy, hyaline
margin at lower half, apex acuminate. Cymule
subsessile, the longest peduncle ranges from
0.5– 1mm long, subtended by 2- unequal tertiary
bracts, the larger one 8– 13x 0.8– 1mm, linearlanceolate, apex sharply acuminate, the shorter
6– 9x 0.8– 1mm, linear-lanceolate, acuminate
apex, pubescent with glandular and sparsely
eglandular hairs and hyaline margin at lower half.
Each cymule composed of 2– (– 3) flowers with 2–
bracteoles each, maturing sequentially. Bracteoles
as bracts, 5– 6x 0.5mm (Fig. 1 B). Calyx whitish
green, sparsely hairy outside, tube 0.5– 1mm
long, lobes lanceolate, 3– 4x 0.5mm attenuated to
sharp acuminate apex and ciliate margin. Corolla
bilabiate, resupinate 8– 8.5mm long, densely
477
eglandular hairy outside, glabrous inside, tube 3–
3.5mm long, lower lip tri-fid 4– 5x 2.5mm, upper lip
retuse to acute 5– 5.5x 3mm (Fig. 1 C). Stamens 2,
epipetalous, filaments 3– 5mm long, sparsely hairy,
anther-thecae superposed, 0.5x 0.3mm. Pollen
grains prolate, medium–sized, with mean polar
axis ranges from 32.1– 35.8µm, mean equatorial
diameter ranges from 20– 21.9µm, circulartriangular in polar view, heterocolpate (tricolporate
with paired pseudocolpi parallel to each colporus),
both colpori and pseudocolpi of about equal length
(30.7– 34.6µm), colpal membrane extremely fine
reticulate, endoapertures raised circular; exine
sculpture foveolate to micro-reticulate (Fig. 2).
Ovary oblong 1.5– 2x 0.5mm, surrounded by a
cupular disc at base, attenuate at apex, eglandular
hairy; style filiform, 6– 7mm long, glabrous,
stigma bi-fid. Capsule 4-seeded, elliptic, 6– 7x
2mm, densely retrorse eglandular multicellular
hairs, with a short rostrum (sterile portion) 1.5–
2mm long. Seeds discoid, 2x 2– 2.2mm, notched
at central white hilum, seed surface variously
convoluted, indistinct reticulation, covered with
minute papillae and tubercles. Tubercles dense
around the edges of the seed, with sharp pointed
hooks in one or two rows (Fig. 3).
Distribution: Confined to the southern coastal
plain of the Red Sea of Egypt.
Specimes examined: Wadi El Faraied, Red
Sea coast 23o31’0’’N; 35o19’60’’E; 12/2/1961;
Täckholm et al. 856 (holotype CAI) – Gebel
Hamata, Red Sea coast, 7/2/1961; Täckholm et al.
351(CAI).
Habitat: Very rare in sandy and silty moist soils.
Etymology: the specific epithet aegyptiaca
refers to Egypt where it was first discovered.
Recognition: Dicliptera aegyptiaca closely
resembles D. paniculata but differs in its stem
unbranched, rarely with short lateral branches
above (vs basely and laterally branched),
Inflorescence congested at the nodes with dwarf
axes 1.5– 5mm long, composed of (1–) 2 – 3
cymules, cymule subsessile (vs inflorescence lax
panicle-like, opposite at each nodes, with long axes
(20–) 30– 50mm long, composed of 3– 4 cymules,
cymule pedunculate). Pollen dimention 32.1–
35.8x 20– 21.9µm, faveolate to micro-reticulate
sculpture (vs pollen dimention 40x 25– 30µm,
reticulate sculpture). (Fig. 1 and 2).
Egypt. J. Bot. 59 , No.2 (2019)
478
EMAN M. SHAMSO AND AHMED S. FOUAD
Fig. 1. Dicliptera aegyptiaca E. Shamso (Täckholm et al. 856); A: Image of specimen showing habit, B: congested
inflorescence and cymule, C: Rusupinate corolla.
Fig. 2. SEM micrographs of pollen grains of: Dicliptera aegyptiaca; A: Whole grain, equatorial view, B: Exine
sculpture.
Egypt. J. Bot. 59 , No.2 (2019)
DICLIPTERA AEGYPTIACA (ACANTHACEAE), A NEW SPECIES...
479
Fig. 3. SEM micrographs of seeds of Dicliptera aegyptiaca; A: Whole seed, B: Testa sculpture.
clade. DNA sequences therefore appear to help
recognize groups of individual specimens and are
useful for species recognition. The paraphyly of
Dicliptera observed in this investigation was also
recognized in other Acanthaceae genera including
Acanthus using ITS sequences (McDade et al.,
2000 b) and Justicia using ITS as well as plastid
trnL-K, trnS-G and rps16 (Deng et al., 2016).
rbcl-based
DNA barcoding
provided
additional support for Dicliptera aegyptiaca that
appeared in the phylogentic tree (Fig. 4), the
tree revealed considerable interspecific variation
between Dicliptera aegyptiaca and D. paniculata.
The new species D. aegyptiaca clearly formed
one clade strongly supported with a bootstrap
value of 100% while D. paniculata along with
other Dicliptera spp. formed a non-sister large
8
KR736771.1 Hypoestes sp.
KX783896.1 Hypoestes phyllostachya
2
KR737526.1 Hypoestes sp.
18
10
KX783894.1 Hypoestes phyllostachya
KX783895.1 Hypoestes phyllostachya
95
10
64
KR736995.1 Hypoestes sp.
L12593.1 Hypoestes forsskaolii
KF496829.1 Hypoestes floribunda
AB586153.1 Hypoestes sp.
88
79
69
AB586152.1 Hypoestes sp.
52
KX397773.1 Dicliptera sexangularis
32
KX783902.1 Dicliptera suberecta
JQ933303.1 Dicliptera obanensis
20
51
AM234782.1 Dicliptera cernua
KF425772.1 Dicliptera paniculata
MH028051 Dicliptera paniculata
64
21
MG199043.1 Dicliptera paniculata
KU947961.1 Dicliptera aegyptiaca
100
KU947958.1 Dicliptera aegyptiaca
KC825342.1 Nicotiana tabacum
Fig. 4. Phylogenetic tree using Maximum Likelihood method based on Tamura 3 parameter model for rbcl
sequences of Hypoestes, Dicliptera and Peristrophe spp. Bootstrap values based on 1000 replications are
listed as percentages at branching points.
Egypt. J. Bot. 59 , No.2 (2019)
480
EMAN M. SHAMSO AND AHMED S. FOUAD
Key to the Egyptian species of Dicliptera
1- Stem unbranched, rarely with short lateral
branches above. Inflorescence congested at the
nodes with short axes, 1.5– 5mm long. Cymule
subsessile, peduncles 0.5– 1mm long, the larger
cymule-bract 8– 13x 0.8 – 1mm, linear-lanceolate,
apex sharply acuminate……..……....D. aegyptiaca
- Stem basely and laterally branched.
Inflorescence in lax panicle- like, with axes (20–)
30– 50mm long. Cymule pedunculate, peduncles
10– 30mm long, the larger cymule-bract 12–
15x 1mm, oblong -linear, apex sharply acute…
..............D. paniculata
Conclusion
Results of the present investigation reflect a
harmony between morphological features and
DNA barcoding in delimiting D. aegyptiaca as
a new species. On the other hand, appearance
of D. aegyptiaca as an outgroup for a large clade
containing taxa of Dicliptera and Hypoestes supports
validity of DNA barcoding in species identification
rather than taxonomy. After the introduction of this
new species to science, the number of Dicliptera
species in Egypt is raised to two.
Acknowledgements: The authors thank Prof Dr.
Kevin Balkwill, University of the Witwatersrand,
Faculty of Science, Johannesburg, South Africa,
for his critical reading, valuable comments and
valuable suggestions on the manuscript. Special
thanks also go to prof. Dr. Hassna Hossni, Cairo
University, Egypt, for revising the manuscript.
References
Abedin, S., Chaudhary, S., Al-yahya, M.A., Mossa, J.S.
and Al Said, M.S. (2000) Acanthaceae, In: "Flora of
the Kingdom of Saudi Arabia" Chaudhary, S. (Ed.),
illustrated, Volume II(3). Ministry of Agriculture
and Water National Herbarium.
Al Hakimi, A., Maideen, H., Saeed, A., Faridah, Q. and
Latiff, A. (2017) Pollen and seed morphology of
Justicieae (Ruellioideae, Acanthaceae) of Yemen.
Flora, 233, 31-50.
Andrews, F.W. (1956) "The Flowering Plants of the
Sudan" (Compositae- Graminae) Volume 3. T.
Buncle & Co., Ltd., Arbroath, Scotland, pp. 163191.
Ardiyani, M., Newman, M.F. and Poulsen, A.D. (2017)
A new species of Zingiber (Zingiberaceae) east of
Wallace’s Line. Gardens’ Bulletin Singapore, 69,
189-199.
Balkwill, K. (1996) A synopsis of Peristrophe
(Acanthaceae) in southern Africa. Bothalia, 26, 8393.
Balkwill, K. and Getliffe Norris, F. (1989) Taxonomic
studies in the Acanthaceae. Peristrophe decorticansA new species. South African Journal of Botany, 55,
254-258.
Balkwill, K., Cadman, M.J. and Getliffe Norris, F.
(1985) Taxonomic studies in the Acanthaceae. New
species of Peristrophe from the Limpopo Valley.
South African Journal of Botany, 51, 485-488.
Balkwill, K., Getliffe Norris, F. and Schoonraad, E.
(1986) Taxonomic studies in the Acanthaceae: Testa
microsculpturing in southern African species of
Peristrophe. South African Journal of Botany, 52,
513-520.
Balkwill, K., Getliffe Norris, F. and Balkwill, M.J.
(1988) Taxonomic studies in the Acanthaceae:
The Peristrophe grandibracteata complex. South
African Journal of Botany, 54, 47-54.
Balkwill, K., Getliffe Norris, F. and Balkwill, M.J.
(1996) Systematic studies in the Acanthaceae;
Dicliptera in southern Africa. Kew Bulletin, 51,
1-61.
Barthlott, W. (1981) Epidermal and seed surface
characters of plants: Systematic applicability and
some evolutionary aspects. Nordic Journal of
Botany, 1, 345-355.
Boulos, L. (2002) "Flora of Egypt" (VerbenaceaeCompositae) Volume 3, pp. 103-104. Al Hadara
Publishing, Cairo.
Boulos, L. (2009) "Flora of Egypt" checklist revised
annotated edition, pp. 214-215. Al Hadara
Publishing, Cairo.
CBOL Plant Working Group (2009) A DNA barcode for
land plants. Proceedings of the National Academy
of Sciences of the United States of America, 106,
12794-12797.
Clarke, C.B. (1901) Acanthaceae, In: "Flora Capensis",
Egypt. J. Bot. 59 , No.2 (2019)
DICLIPTERA AEGYPTIACA (ACANTHACEAE), A NEW SPECIES...
Thiselton-Dyer, W.T. (Ed.), pp. 1-92. Volume 5(1).
Reeves & Sons Pty, Ltd, London.
Costea, M. and Stefanović, S. (2009) Molecular
phylogeny of Cuscuta californica complex
(Convolvulaceae) and a new species from New
Mexico and Trans-Pecos. Systematic Botany, 34,
570-579.
Daniel, T.F. (2009) Synopsis of Dicliptera (Acanthaceae)
in the Nueva Galicia region of western Mexico with
a new species, D. novogaliciana. Proceedings of the
California Academy of Sciences, 60, 17-18.
Darbyshire, I. and Vollesen, K. (2007) The transfer of
the genus Peristrophe to Dicliptera (Acanthaceae),
with a new species described from eastern Africa.
Kew Bulletin, 62, 119-128.
Darbyshire, I., Vollesen, K. and Ensermu Kelbessa
(2010) Acanthaceae. In: "Flora of Tropical East
Africa", Beentje, H.J. (Ed.), part 2. Richmond:
Royal Botanic Gardens, Kew.
Darbyshire, I., Vollesen, K. and Ensermu Kelbessa
(2015) Acanthaceae. In: "Flora Zambesiaca",
Timberlake, J.R. and Martins, E.S. (Eds.), Volume
8(6). Richmond: Royal Botanc Gardens, Kew.
Deng, Y., Gao, C., Xia, N. and Peng, H. (2016)
Wuacanthus (Acanthaceae), a new Chinese endemic
genus segregated from Justicia (Acanthaceae).
Plant Diversity, 38, 312-321.
de Vere, N., Rich, T.C.G., Ford, C.R., Trinder, S.A.,
Long, C., Moore, C.W., Satterthwaite, D., Davies,
H., Allainguillaume, J., Ronca, S., Tatarinova, T.,
Garbett, H., Walker, K. and Wilkinson, M.J. (2012)
DNA Barcoding the native flowering plants and
conifers of Wales. PLoS ONE, 7, e37945.
El-Gazzar, A., Khafagi, A.A., El-Husseini, N. and
Mostafa, N.A. (2015) Computer-generated keys to
the flora of Egypt. 7. The Acanthaceae s.l. Annals of
Agriculture Science, 60(2), 257-277.
El Hadidi, M.N. and Fayed, A.A. (1994/95) Materials
for excursion Flora of Egypt. Taeckholmia, 15, 147.
Ensermu Kelbessa (2003) Two new species of
Acanthaceae from NE tropical Africa and Arabia.
Kew Bulletin, 58, 703-712.
Ensermu Kelbessa (2006) Acanthaceae, In: "Flora
481
of Ethiopia & Eritrea. Gentianaceae to
Cyclocheilaceae", I. Hedberg, Ensermu Kelbessa, S.
Edwards, Sebsebe Demissew and E. Persson (Eds.),
pp. 345-495. Vol. 5. The National Herbarium, Addis
Ababa University, Ethiopia and The Department of
Systematic Botany, Uppsala, Sweden.
Felsenstein, J. (1985) Confidence limits on phylogenies:
An approach using the bootstrap. Evolution, 39,
783-791.
Hebert, P.D.N. (2003) Biological identifications through
DNA barcodes. Proceedings of the Royal Society of
London B Biological Sciences, 270, 313-321.
Hebert, P.D.N. and Gregory, T.R. (2005) The promise of
DNA barcoding for taxonomy. Systematic Biology,
54, 852-859.
Hedrén, M. (2006) Acanthaceae, In: "Flora of Somalia"
M. Thulin (Ed.). Volume 3. Royal Botanic Gardens,
Kew.
Heine, H. (1966) Acanthaceae. In: "Flore du Gabon".
pp. 3-250. Volume 13, Paris: Museum National
d’Histoire Naturelle.
Hesse, A., Halbritter, H., Zetter, R., Weber, M., Buchner,
R., Andrea, F. and Ulrich, S. (2009) "Pollen
Terminology". An illustrated Handbook. Springer
Wein, New York.
Kang, Y., Deng, Z., Zang, R. and Lon, W. (2017) DNA
barcoding analysis and phylogenetic relationships
of tree species in tropical cloud forests. Scientific
Reports, 7, 12564.
Kiel, C.A., Daniel, T.F., Darbyshire, I. and McDade,
L.A. (2017) Unraveling relatioships in the
morphologically diverse and taxonomically
challenging “justicioid” lineage (Acanthaceae:
Justicieae). Taxon, 66, 645-674.
Liu, Y.J., Newmaster, S.G., Wu, X.J., Liu, Y., Ragupathy,
S., Motley, T. and Long, C.L. (2013) Pinellia
hunanensis (Araceae), a new species supported
by morphometric analysis and DNA barcoding.
Phytotaxa, 130, 1-13.
McDade, L.A., Daniel, T.F., Masta, S.E. and Riley, K.M.
(2000 a) Phylogenetic relationships within the tribe
Justicieae (Acanthaceae): evidence from molecular
sequences, morphology and Cytology. Annals of the
Missouri Botanical Garden, 87, 435-458.
Egypt. J. Bot. 59 , No.2 (2019)
482
EMAN M. SHAMSO AND AHMED S. FOUAD
McDade, L.A., Masta, S.E., Moody, M.L. and Waters,
E. (2000b) Phylogenetic relationships among
Acanthaceae: evidence from two genomes.
Systematic Botany, 25, 106-121.
Miwa, H., Suhara, J., Kitagawa, N. and Murakami,
N. (2003) Biosystematic study of Japanese
Conocephalum japonicum (Hepaticae) based on rbcl
sequence and allozyme data. Acta Phytotaxonomica
et Geobotanica, 54, 37-48.
Newmaster, S.G., Fazekas, A.J.J. and Ragupathy, S.
(2006) DNA barcoding in land plants: Evaluation
of rbcl in a multigene tiered approach. Canadian
Journal of Botany, 84, 335-341.
Shamso, E.M. (2013) A Palynological study of
Acanthaceae in Egypt and its systematic implication.
Egyptian Journal of Botany, 53, 257-272.
Täckholm, V. (1974) "Students’ Flora of Egypt". 2nd ed,
Cairo University.
Tamura, K. (1992) Estimation of the number of
nucleotide substitutions when there are strong
transition-transversion and G+C-content biases.
Molecular Biology and Evolution, 9, 678-687.
Tamura, K., Stecher, G., Peterson, D., Filipski, A. and
Kumar, S. (2013) MEGA6: Molecular evolutionary
genetics analysis version 6.0. Molecular Biology
and Evolution, 30, 2725-2729.
Shamso, E.M. (2010) A Synoptic revision of
Acanthaceae in Egypt, with a new record from
Gebel Elba. Taeckholmia, 30, 127-144.
(Received 4/10/2018;
accepted 12/3 /2019)
نوع جديد من مصر مدعوما ً بالصفات،) (العائلة األكانثيةDicliptera aegyptiaca
rbcl المورفولوجية و تقنية الباركود لألحماض النووية باستخدام جين
أحمد فؤاد،(1)إيمان شمسو
، مصر- (المعشبة – قسم النبات والميكروبيولوجي – كلية العلوم – جامعة القاهرة – الجيزة1)
.والميكروبيولوجى – كلية العلوم – جامعة القاهرة – الجيزة – مصر
(2)
قسم النبات
(2)
كنوع جديد بمنطقة ساحل البحر األحمر بمصر وقد تناولDicliptera aegyptiaca أسفرت الدراسة عن وجود
Dicliptera paniculata البحث دراسة الصفات المورفولوجية والصفات التمييزية بينه و بين النوع القريب له
.rbcl DNA مع عمل مفتاح تعريفي لهما و تم عمل باركود للنوع الجديد ألول مرة باستخدام الشفرة الوراثية للجين
مللي طول5-1,5 بنبات ذو ساق غير متفرع و نورة ذات محور متقزمDicliptera aegyptiaca يتميز النوع.
أظهرت الشجرة التطورية للشفرة الوراثية لهذين النوعين. مللي طول1-0,5 و نويرة محدودة ذات شمراخ قصير
كفرع منفرد مدعوما بتكررات تأكيدDicliptera aegyptiaca وجود فروق بينهما وهذا اتضح من ظهور
وقد أظهرت الدراسة المورفولوجية للنبات و لحبوب اللقاح و البذور إنتماء النبات الجديد لجنس.100% بقيمة
بينما اظهرت الشجرة التطورية النبات الجديد فى فرع آخر غير المخصص لباقى أنواع الجنس مماDicliptera
.يظهر فائدة تقنية الباركود فى تعريف األنواع أكثر من إيجاد العالقات التصنيفية بينها
Egypt. J. Bot. 59 , No.2 (2019)