A COMPARATIVE SYSTEMATIC STUDY OF THE GENUS Fritillaria L. (Liliaceae) IN IRAQI KURDISTAN REGION
A thesis Submitted to the Council of the Faculty of Agricultural Sciences at the University of Sulaimani in partial fulfillment of the requirements for the degree of Master of Science in
Horticulture - Plant Taxonomy
By
Lanja Hiwa Mustafa Khal B.Sc. Horticulture (2007), University of Sulaimani
Supervised by
Dr. Rupak Towfiq Abdul-Razaq Lecturer
2713 K.
2013 A. D
بسم اه الرمن الرحيم َيزِفَعِ الَلهُالَ ِذينَ آمَنُوامِنِ ُكِم وَالَ ِذينَأُوتُواالِعِِلَم ٌ ن خَبر َ ت وَالَلُهِبمَاتَِعَملُو ٍ دَرَجَا صدق اه العظيم ]11[ اجادلة
In the name of Allah, the Beneficent, the Merciful
Allah will exalt in degree those of you who believe and those who have been granted knowledge. And Allah is Well-Acquainted with what you do.
Allah told the Truth Al-Mujadala ]11[
SUMMARY The current work includes a comparative systematic study of the taxa relating to the genus Fritillaria L. family (Liliaceae). Morphological, palynological, anatomical characters as well as molecular study, Ecological and geographical distribution study and taxonomic treatments were included. The morphological study was dealed with studying the characters of bulbs, stems, leaves, flowers, inflorescence, fruit parts and seeds. The diversity in characters was discussed and it was indicated that the flowers characters are more important taxonomically than vegetative characters to isolate the species. This study indicates that pollen grains of all the taxa of the genus Fritillaria under the study are monads and referred to that the characters of pollen grains have a taxonomic importance. The present study also includes the anatomy of leaf epidermis, main vein of leaf, stomata distribution in the leaf and cross sections of each the leaf and stem. For the first time in Iraq, it was indicated that some anatomical characters have taxonomical importance in identifying the species within this genus. The present study reported a molecular method based on the sequences of nuclear ribosomal DNA internal transcribed spacer (nrDNA ITS) regions (18S rDNA). For identification of Fritillaris spp. a pair of species-specific primers was designed to authenticate Fritillaria spp. Sequence analysis tools were established to compare and differentiate Fritillaria local isolates from the other species of Fritillaria. This method provides an effective and accurate identification of Fritillaria spp. Ecological and geographical study don for all the species which grows in Iraqi Kurdistan region. I
Summary
As a result of the study one sub species were recorded to the first time in Iraq which is F. alfredae subsp. glaucoviridis. Depending on the data concluded from macro and micro-morphological characters, molecular data had also been studied due to their importance and occurrence in identifying the species, and a taxonomic key was prepared depending on macro morphological traits of species of the studied genus. The taxa which investigation in this study are Fritillaria imperialis, F. persica, F. assyriaca, F. uva-vlpis, F. crassifolia subsp. kurdica, F. crassifolia subsp. hakkarensis, F. alfreda subsp. glaucoviridis .
II
ثـــــوخـــتة لةم تويَريهةوةيةدا ليَكوَلَيهةوةيةكي ثوَليَهكسدني بةزاوزدكةزي يةمةايةنةية لةضةز زِةطةشي Fritillariaضةزبة خيَصاني Liliaceaeكة ثشي بةضتووة بةايةني زِووكازي (موَزفوَلَوَجي) ,دةنكة يةاَلَة, خةضمَةتةكاني تويَكازي و ليَكوَلَيهةوةي طةزديمةيي ضةزةزِاي مامةلَةي ثوَليَهكسدى. ليَكوَلَيهةوةي تويَكازي ثيَكًاتووة لة ليَكوَلَيهةوةي خةضمَةتي ضةلك ,قةد ,طةاَ ,طولَ ,طولَ كوَ ,بةز وة توَو .لةئةجامدا دةزكةوت ,خةضمَةتةكاني طولَ شوَزتسيو طسنكي يةية لة ثوَليَهكسدى بة بةزاوزد لةطةلَ خةضمَةتةكاني بةشة ضةوشةكاى بوَ جياكسدنةوةي جوَزةكاى .يةزوةيا ئةم ليَكوَلَيهةوةية ئاماذة دةدات بةوةى كة دةنكة يةاَلَةكاني جوَزي زِةطةشي Fritillariaكة ليَكوَلَيهةوةي لةضةز كساوة لةجوَزي يةك زِووى وة خةضمَةتةكاني دةنكة يةاَلَة طسنكي يةية لة ثوَليَهكسدندا. ئةم ليَكوَلَيهةوةية ئاماذة دةدات بة تويَكازي زِووثوَشة خانةي طةاَ ,دةمازي ناوةزِاضت لة طةاَ, باَوبوونةوةي جةميمة لةضةز زِووي طةاَ وة ئامادةكسدني ثانة بسِطة لة قةد و طةاَدا بوَ يةكةم جاز لة عراقدا, ئاماذة دةدات بةوةي كة يةنديَك لة خةضمَةتةكاني تويَكازي طسنكي ثوَليَهكسدني يةية بوَديازيكسدني جوَزةكاني ئةم زِةطةشة . لةم ليَكوَلَيهةوةيةدا ضوود لةزِيَطاي طةزديمةيي وةزطراوة لةضةز بهةماي شجرةي ناوكي بوَ ديازيكسدني شويَي ) (nr DNA ITSلة جياتي ) (18SrDNAبوَديازيكسدني جوَزةكاني زِةطةشي , Fritillariaبة بةكازييَهاني ئاميَسةكاني شيكسدنةوةي شجرةي ئامادةكساو بوَ بةزاووزدكسدى و جياكسدنةوةي Fritillariaي
ناوخوَ وة جياكسدنةوةياى لة جوَزةكاني تسي .Fritillaria ليَكوَلَيهةوةي ذيهطةي بوَ يةموو جوَزةكاني ئةم زِةطةشة ئةجام دزا .لة ئةجامي ئةم ليَكوَلَيهةوةيةدا F.alfredae subsp. glaucoviridisبوَ يةكةم جاز لة عراقدا توَماز كسا .لةم ليَكوَلَيهةوةيةدا كميمي ثوَليَهكسدى ئامادةكسا بة ثشت بةضن بة خةضمَةتةكاني زِووكازي طةوزة بوَ جوَزةكاني زِةطةشي Fritillariaلةم تويَريهةوةيةدا .ئةو جوَزانةي لةم تويَريهةوةيةدا ليَكوَلَيهةوةي لةضةز كساوة بسيي ية لة Fritillaria imperialis, F. persica, F. assyriaca, F. uva-vlpis, F. crassifolia subsp. kurdica, F. crassifolia subsp. hakkarensis, F. alfreda subsp. glaucoviridis. أ
اخـــاصـــــــــُ يتطنن البخث احالٌ دراصُ تصنًفًُ مقارنُ مزاتب اجنط Fritllariaالتابعُللعآلُ الشنبقًُ Liliaceaeمن النىاحٌ امظوزيُ و حبىب اللقاح و الصفات التشزحًُ والدراصُ اجشئًُ فطا" عن اجزاْ امعاملُ التصنًفًُ. تناولت الدراصُ امظوزيُ صفات كل من البصلُ والضاق و ااْوراق و ااْسهار و النىرَ الشهزيُ و الثنار والبذور .وتبن ان صفات الشهزَ هٌ ااْكثز اْهنًُ من الناحًُ التصنًفًُ من الصفات اخطزيُ لعشل ااْنىاع. بًنت هذه الدراصُ ان حبىب اللقاح جنًع ااْنىاع امدروصُ جنط Fritllariaهٌ وحًدَ امظوز و ان صفات حبىب اللقاح ها اْهنًُ تصنًفًُ. تشنل الدراصُ احالًُ اْيظا"النىاحٌ التشزحًُللىرقُ ( البشزَ و تىسيع الثغىر وعنل مقاطع متعزضُ للل من الضاق والىرقُ.وتبن ان بعض الصفات التشزحًُ ها اْهنًُ تصنًفًُ ي تشدًص ااْنىاع داخل اجنط قًد الدراصُ. من جوُ اْخزّ افادت هذه الدراصُ اى الطزيقُ اجشئًُ امبنًُ علِ اصاظ التضلضل النىوٍ ()DNA لتخديد ) nr DNA ITSمىقع ). (18SrDNAتشدًص ااْنىاع اجنط Fritllariaالنىع امذدوج اخاص من بزامز مت التصنًه لتصديق . Fritllaria sp.ادوات التخلًل م تاْصًضوا حلًاّ للنقارنُ والتنًًش اْنىاع اجنط Fritllariaوعشها عن بعطوا البعض .هذه الطزيقُ منح الفعالًُ والدقُ لتشدًص انىاع اجنط .Fritllaria خال الدراصُ اجزيت الدراصُ البًًُٔ جنًع انىاع اجنط قًد الدراصُ ,كنا صجلت النىيع F.alfredae subsp. glaucoviridisاْول مزَ ي العزاق .وم تثبًت مفتاح تصنًفٌ لاْنىاع قًد الدراصُ اعتنادا" علِ الصفات امظوزيُ امعتند ي التشدًص. امزاتب الي مت دراصتوا خال هذه الدراصُ هٌ Fritillaria imperialis, F. persica, F. assyriaca, F. uva-vlpis, F. crassifolia subsp. kurdica, F. crassifolia subsp. hakkarensis, F. alfreda subsp. glaucoviridis.
أ
Acknowledgment My infinite thanks to Allah, who made me sound and patient in completing this project. My sincere thanks and appreciations to the presidency of Sulaimani University and particular the Deanery of the Faculty of Agriculture Sciences and Horticulture Department for giving me the opportunity to complete my study. I would like to thanks all the staff of Faculty of Agriculture Sciences, my gratitude is particularly due to Assist. Prof. Dr. Aram A. Muhammad, the Faculty Dean, for his effort, encouragement and facilities which made this task to be performed. I would also like to present my thanks to the Head of Horticulture Department, Dr. Hama Rasool S. Rasool, for supporting and providing the facilities of the study. Special words of thanks and appreciations are due to Dr. Rupak T. Abdul-Razaq, for her kind supervision, scientific support and guidance in completing this work. I would like to express my sincere thanks and gratitude to respectable Dr. Dlnia A. Mohammed and Dr. Shwan M. Salh, for their untiring assistances during my time working with this research. Grateful recognition also to Prof. Dr. Athiya N. Salman, Assist. Prof. Dr. Jawher Fattah Saeed, Assist. Prof. Dr. Nawroz A. R. Tahir, Dr. Ibrahim M. Noori, MR. Ali Mala Galalaey, Miss. Ashtyxwaz A. Shareef, Mr. Hoshman O. Majeed, Mr. Faraidoon K. Ahmad, Mr. Faraidoon M. H. Faraj, Mr. Zeravan A. Sadiq and Mr. Abdul-Aziz S. Abdul-Muhseen. Many thanks are due to my collages and friends especially {Kochar, Hawar, Mrs. Zhalla, Miss. Ruxosh, Miss. Sonia, Mrs. Hazha, Mrs. Kana, Awara, Parwez and Amina}. To all my family, you are my constant source of inspiration, thank you for understanding and not resenting me for the decision I have made which have taken me away from you. I think of you all every day and love you with all my heart. Today and always, thank you for everything. BEST REGARDS Your's sincerely
Lanja H. Mustafa Khal
Appendix (1): List species of the genus Fritillaria that mentioned by HandelMazzetti (1910) in Iraqi Kurdistan region. No.
Species
District
1.
Fritillaria imperialis
Su
2.
F. persica
3.
F. assyariaca
A
4.
F. kurdica
A
5.
F. crassifolia
Su
Hult
Appendix (2): List species of the genus Fritillaria that mentioned by Guest (1933) in Iraq. No.
Species
District
1.
Fritillaria imperialis
Common on mountain slopes in Kurdistan
2.
F. persica
3.
F. assyariaca
Amadian
4.
F. crassifolia
Sulaimanian
Rowanduz
Appendix (3): List species of the genus Fritillaria that mentioned by Zohary (1946) in Iraq. No.
Species
District
1.
Fritillaria imperialis
Su
2.
F. persica
A
3.
F. assyariaca
A
4.
F. kurdica
A
5.
F. crassifolia
Su
113
Appendix (4): List species of the genus Fritillaria that mentioned by Al-Rawi (1964) in Iraq. No.
Species
District
1. 2. 3. 4.
Fritillaria imperialis F. persica F. assyariaca F. kurdica
A, R, SU R, Su, Si A, Su Su
5.
F. crassifolia
Su
6.
F. olivieri
A
Appendix (5): List species of the genus Fritillaria that mentioned by Ridda and Daoud (1982) in Iraq. No.
Species
District
1.
Fritillaria imperialis
MSU, MRO, MAM
2.
F. persica
3.
F. assyariaca
MSU, MRO, MAM
4.
F. uva-vulpis
MSU, MRO, MAM
5.
F. crassifolia
MSU,MRO, MAM
MSU, MRO, FKI, MJS
Appendix (6): Abbreviations of Iraqi Herbaria names which mentioned in this study (cited from Holmgren and Keuken, 1964). No. Abbreviations 1.
SUH
2.
DUH
3.
BAG
Herbaria names Erbil, Iraq: College of Science, University of Salahaddin. Duhok, Iraq: College of Agriculture, University of Duhok. Baghdad, Iraq: The National Herbarium, College of Science. 114
115
LIST OF CONTENTS Title
Series Summary
Page No. I
List of contents
III
List of figures
III
List of tables
III
List of diagrams
X
CHAPTER ONE: INTRODUCTION AND LITERATUR E REVIEW
1.1
General introduction
1
1.2
Literature review
4
1.3
Taxonomic status of the genus Fritillaria
1.4
Taxonomic status of the genus Fritillaria in Iraq Origin of the name Fritillaria and its colloquial
8
names
8
1.6
Common and medicinal uses
9
1.7
Fritillaria side effects
11
1.5
5
CHAPTER TWO: MORPHOLOGICAL STUDY 2.1
Introduction
12
2.2
Materials and methods
13
2.3
Results and discussion
14
2.3.1
Habit and duration
2.3.2
Bulbs
2.3.3
Stems
2.3.4
Leaves
18
2.3.5
Flowers
18
14 14 14
III
Series 2.3.6
Title
Page No.
Androecium
26
2.3.6.1
Filament
26
2.3.6.2
Anther
2.3.7
26
Gynoecium
29
2.3.7.1
Stigma
29
2.3.7.2
Style
29
2.3.7.3
Ovary
3.8
Fruit
29
2.3.9
Seed
30
29
CHAPTER THREE: PALYNOLOGICAL STUDY 3.1
Introduction
34
3.2
Materials and methods
35
3.3
Results and discussion
36
CHAPTER FOUR: ANATOMICAL STUDY 4.1
Introduction
41
4.2
Materials and methods
43
4.2.1
Epidermis and stomata
4.2.2 4.2.2.1
Slide preparation Killing and fixation
4.2.2.2
Washing and dehydration
4.2.2.3
Clearing and infiltration
4.2.2.4
Embedding and mounting
43 43 43 43 44 44
IV
Series Title 4.2.2.5 Removing of wax and staining 4.3
Result and discussion
4.3.1
Stem anatomy
4.3.2
Leaf anatomy
Page No. 45 45 45 48
4.3.2.1 Epidermis
48
4.3.2.2 Mesophyll tissue
51
CHAPTER FIVE: MOLECULAR STUDY 5.1
Introduction
53
5.2
Materials and methods
55
5.2.1
Plant material
5.2.2
Apparatus and equipment
5.2.3
Chemical compounds
55 55 56
5.2.4
Molecular marker methods 5.2.4.1 DNA extraction from Fritillaria spp.
56
5.2.4.2 Protocol for DNA extraction
56
5.2.4.3
56
Amplification of Fritillaria spp. 18S- ribosomal RNA 57
intergenic region by PCR 5.2.4.4 Visualization of PCR products
58
5.2.4.5 Fritillaria spp. 18Sr gene sequencing and sequence 58
alignment 5.3
Results and discussions
58
V
Series
Title
Page No.
CHAPTER SIX: ECOLOGICAL AND GEOGRAPHICAL STUDY
6.1
Introduction
64
6.2
Materials and methods
66
6.3
Results and discussion
67
6.3.1
Ecological notes
6.3.2
Geographical distribution
6.3.3
Phenology
67 73 76
CHAPTER SEVEN: GENERAL DISCUSSION
79
CHAPTER EIGHT: TAXONOMICAL TREATMENTS AND KEY Keys
86
Taxonomical treatment
87
CONCLUSIONS AND RECOMMENDATIONS
99
REFFERENCES
VI
100
LIST OF FIGURES Fig. No.
Figure title
Page No.
Variation in the shapes and dimensions of bulb of the (1)
16
genus Fritillaria taxa. Variation in the shapes and dimensions of leaf of the
(2) (3)
19
genus Fritillaria taxa. Inflorescence of all investigated species.
22
Variation in the shapes and dimensions of fruit of the (4)
32
genus Fritillaria taxa. Variation in the shapes and dimensions of seed of the
(5)
33
genus Fritillaria taxa. Variations in the shapes and dimensions of pollen
(6)
grain of Polar view of the genus Fritillaria taxa.
39
Variations in the shapes and dimensions of pollen (7) (8) (9)
grain of equatorial view of the genus Fritillaria taxa. Cross secition of stem of all investigated taxa. Epidermis cell of all investigated taxa.
40 47 49
Type of stomata of all investigated taxa in adaxial (10)
surface.
49
(11)
Cross secition of leaf of all investigated taxa.
52
Gel electrophoresis of the amplified PCR product of the 18S rDNA gene.Lane 1: 100 bp DNA ladder, (12)
lane 2, 3, 4, 5, 6, 7: 600-700 bp 18S rDNA of Fritillaria sp.
VII
58
Fig. No.
Figure title
Page No.
Alignment of 18S rDNA of Fritillaria isolate No.1 with Fritillaria imperialis 18S rRNA gene, partial (13)
sequence; internal transcribed spacer 1, 5.8S r RNA
59
gene (AY616725.1), identity=389/407 (96%). Alignment of 18S rDNA of Fritillaria isolate No. 2 (14)
with Fritillaria persica (AY616736), identity=
60
402/422 (96%). (15)
Alignment of 18S rDNA sequence of Fritillaria isolate No. 3 with Fritillaria raddeana
61
(AY616739.1), identity= 90%. (16)
Distribution of 100 Blast Hits on the Query Sequence
62
(17)
Physiographic sketch map of Iraq
69
Map of Northern Iraq Kurdistan region showing the (18) (19)
distribution of the genus Fritillaria
70
Map of Northern Iraq Kurdistan region showing the distribution of the genus Fritillaria
71
Photographic picture of species the genus Fritillaria (20)
72
in natural ecology.
(21)
Number of species which distributed in each district
75
(22)
Flowering period for different species of Fritillaria.
77
(23)
Fruiting period for different species of Fritillaria.
78
VIII
LIST OF TABLES Table No.
Table title
Page No.
Characters of bulb, stem and blade for the taxa (1)
belonging to the genus under the study.
15
Characters of peduncle, nectar and tepal for the taxa (2)
belonging to the genus under the study.
21
Characters of ovary, style, filaments and anthers for the (3)
taxa belonging to the genus under the study, measured
27
by (mm). Characters of fruit and seed for the taxa belonging to (4)
the genus under the study, measured by (mm).
31
Quantitative and qualitative characters of pollen grains (5)
for genus Fritillaria measured by (μm).
37
(6)
List of apparatus used in the study.
55
(7)
List of chemicals used in the study.
56
Distribution of genus Fritillaria of Iraqi Kurdistan (8)
(9)
Region districts, altitudes and types of soil. Distribution and altitude of Fritillaria in Iraqi districts.
IX
68
75
LIST OF DIAGRAMS Diagram No.
Diagram title
Page No.
Variations in the shapes and dimensions of bulbs of (1)
the genus Fritillaria taxa.
17
Variations in the shapes and dimensions of leaves of (2)
the genus Fritillaria taxa.
20
Variations in the shapes and dimensions of tepals of (3)
the genus Fritillaria taxa.
24
Variations in the shapes and dimensions of androecia (4)
and gynoecia of the genus Fritillaria taxa.
28
A- Epidermis cell of all investigated taxa. (5)
B- Type of stomata of all investigated taxa in adaxial surface.
X
50
Morphological Study Table (1): Characters of bulb, stem and blade for the taxa belonging to the genus under the study.
Bulb dimensions (mm) No.
Taxa Length
Diameter
Stem length (cm)
Leaf dimensions (cm) Length
Width
1.
Fritillaria imperialis L.
42.88 - 63.21 (50.03)*
53.30 - 67.83 (60.45)*
60.0 – 126.0 (91.13)*
16.4-17.6 (17.1)*
6.0-8.2 (7.5)*
2.
F. persica L.
42.77 - 64.85 (56.86)*
36.06 - 55.87 (44.56)*
30.0 -95.0 (63.54)*
10.2-15.0 (14.95)*
1.5-3.0 (2.45)*
3.
F. assyriaca Bak.
17.34 - 18.19 (17.64)*
20.94 - 25.6 (23.12)*
10.0 – 29.0 (20.51)*
4.0-9.0 (7.87)*
0.5-1.1 (0.84)*
4.
F. uva-vlpis Rix.
16.2 - 18.31 (17.11)*
11.6 -13.6 (12.40)*
5.0 – 24.0 (15.52)*
3.0-10.0 (7.5)*
0.6-1.2 (0.9)*
14.1 -15.2 (14.35)*
6.9 - 9.2 (7.05)*
3.0 – 9.0 (6.31)*
3.2-7.8 (5.00)*
0.6-1.4 (1.11)*
13.15 - 14.20 (13.55)*
5.65 - 9.17 (6.91)*
7.0 – 17.0 (11.68)*
4.8-10.0 (6.83)*
0.9-2.0 (1.62)*
20.2-22.2 (21.33)*
25.0-35.0 (29.32)*
10.0-35.0 (14.54)*
4.0-10.0 (7.0)*
0.6-3.2 (1.89)*
5. 6. 7.
F. crassifolia subsp. kurdica Boiss. & Noe. F. crassifolia subsp. hakkarensis Rix. F. alfreda subsp. glaucoviridis Rix.
(*) The numbers in the brackets indicate the averages.
15
Morphological Study Table (2): Characters of peduncle, nectar and tepal for the taxa belonging to the genus under the study.
Pedicle (mm) NO.
1. 2.
Nectar (mm)
(Taxa)
Length
Width
Length
Color
Fritillaria imperialis
30.0-33.0 (32.5)*
2.0-4.0(3.21)*
70.0-85.0(79.5)*
White
F. persica
19.0-24.0 (21.6)*
0.09-0.15(0.11)*
1.8-3.7(2.8)*
Black
3.
F. assyriaca
24.0-27.0 (25.39)*
0.9-1.5(1.3)*
2.0-4.0(3.2)*
Black
4.
F. uva-vulpis
22.0-28.0 (25.4)*
1.2-1.9(1.45)*
1.5-3.6(2.4)*
Black
5.
F. crassifolia subsp. kurdica
25.0-27.0 (26.1)*
1.1-1.7(1.53)*
8.0-10.0 (9.1)*
Black
6.
F. crassifolia subsp. hakkarensis
7.
F. alfreda subsp. glaucoviridis
23.0- 25.0 (24.22)*
24.0-26.0 (25.21)*
1-1.5(1.35)*
1.0-1.6 (1.2)*
9.0-11.0 (9.8)*
4.2-6.0 (5.4)*
Green usually with a black spot at the base
Tepal (cm) Length
Width
Outer
5.4-6.5(5.8)*
Outer
1.4-1.8 (1.6)*
Inner Outer Inner
5.4-6.5(5.8)* 2.0-2.4(2.1)* 2.0-2.4(2.1)*
Inner Outer Inner
2.4-2.8 (2.7)* 0.6-0.8 (0.7)* 0.8-1.1(0.9)*
Outer
1.9-2.2(2.0)*
Outer
0.3-0.5 (0.42)*
Inner
1.9-2.2(2.05)*
Inner
0.5-0.7 (0.6)*
Outer
0.4-0.5(0.44)*
Outer
0.6-0.8(0.7)*
Inner
0.5-1.0(0.87)*
Inner
0.6-0.8(0.7)*
Outer
1.6-2.2(1.8)*
Outer
0.5-0.8(0.65)*
Inner
1.6-2.2(1.8)*
Inner
0.8-1.3(1.05)*
Outer
1.5-1.7(1.5)*
Outer
0.4-0.6(0.5)*
Shape Broadly campanulate
1.5-1.7(1.5)*
Inner
0.6-0.7 (0.64)*
Outer
0.6-0.8(0.6)*
Outer
2.1-3.0(2.65)*
Inner
0.7-0.9(0.81)*
Inner
2.2-3.1(2.7)*
Green
(*) The numbers in the brackets indicate the averages.
21
Red, yellow and brownish orange
Ellipitic to obovate
Dark plum-purple and Yellow
Inner linearlanceolate outer Ellipitic to obovate
Greenish or purplish-brown color with green stripes outside, greenish or yellow inside
Inner obovate outer Ovate-lanceolate
Purplish-grey edged yellow outside and deep yellow inside
Obtuse
Greenish or yellow ground with green strips
Acute Inner
Color
Obtuse
Greenish-yellow stripes tessellated inside
Outside glaucous green, inside yellowish-green
Morphological Study Table (3): Characters of ovary, style, filaments and anthers for the taxa belonging to the genus under the study, measured by (mm).
Ovary
Filament
(Taxa)
No.
Anther
Style Length Length
Width
Length
Width
Length
Width
16.0-18.00 (17.2)*
3.0-4.0 (3.6)*
31.0-54.0 (46.5)*
47.0-64.0 (56.5)*
3.0-5.0 (4.1)*
6.0-11.0(8.9)*
3.0-4.0 (3.4)*
6.3-7.5 (6.2)*
2.0-3.0 (2.4)*
17.0-19.0 (18.3)*
6.0-9.8 (8.5)*
1.0-2.0 (1.6)*
2.0-3.0 (2.6)*
1.0-2.0 (1.6)*
1.
Fritillaria imperialis
2.
F. persica
3.
F. assyriaca
11.0-13.0 (12.3)*
3.0-4.0 (3.4)*
6.0-9.0 (8.5)*
6.0-10.0 (8.3)*
1.0-2.0 (1.6)*
4.0-7.0 (5.9)*
2.0-4.0 (3.2)*
4.
F. uva-vulpis
10.0-13.0 (11.4)*
2.0-4.0 (3.2)*
6.0-8.0 (7.2)*
6.0-9.0 (7.8)*
1.0-2.0 (1.7)*
4.0-7.0 (5.9)*
2.0-4.0 (3.22)*
5.
F. crassifolia subsp. kurdica
10.0-13.0 (11.7)*
2.0-3.0 (2.5)*
4.9-8.2 (6.7)*
5.0-8.0 (7.2)*
1.0-2.0 (1.4)*
4.0-7.0(5.4)*
2.0-3.0 (2.4)*
6.
F. crassifolia subsp. hakkarensis
5.0-7.0 (6.4)*
2.0-5.0 (3.4)*
5.0-8.0 (6.3)*
5.0-7.0 (6.4)*
0.9-1.0 (0.85)*
3.0-4.5(3.8)*
2.0-4.0 (3.1)*
7.
F. alfredae subsp. glaucoviridis
5.0-7.2 (6.4)*
2.1-5.1(3.4)*
5.0-10.0(8.5)*
8-10(9.5)*
1.2-2.2(1.66)*
4.0-7.2(6.1)*
2.0-4.0(3.2)*
(*) The numbers in the brackets indicate the averages.
27
Morphological Study Table (4): Characters of fruit and seed for the taxa belonging to the genus under the study, measured by (mm).
Fruit No.
Seed
(Taxa) Length
width
Shape
Length
width
Shape
1.
Fritillaria imperialis
25.0-48.0 (37.5)*
17.0-38.0 (28.5)*
Oblong to cylinder
7.0-10.0(8.7)*
5.0-8.0(6.8)*
Ovateoblong
2.
F. persica
14.0-45.0 (29.8)*
7.3-8.4 (7.9)*
Obconical
5.0-6.0 (5.6)*
3.5-5.0 (4.1)*
Ovate
3.
F. assyriaca
25.0-42.0 (34.5)*
16.0-28.0 (24.0)*
Oblong-ellpisod
6.0-8.0 (7.1)*
3.0-5.0 (4.4)*
Ovate
4.
F. uva-vulpis
25.0-35.0 (31.0)*
7.0-14.1 (11.65)*
Oblong-ellpisod
4.0-6.0 (5.1)*
3.4-5.0 (4.3)*
Ovate
5.
F. crassifolia subsp. kurdica
25.0-30.0 (26.5)*
6.0-13.7 (10.75)*
Cylindrical
5.0-6.0(5.6)*
3.2-5.0 (4.0)*
Ovate
6.
F. crassifolia subsp. hakkarensis
27.0-29.0 (28.2)*
6.6-13.9 (11.35)*
Cylindrical
4.5-6.0 (5.4)*
3.0-6.0 (4.7)*
Ovate
7.
F. alfredae subsp. glaucoviridis
26.0-30.0 (27.2)*
6.5-13.5 (11.33) *
Cylindrical
4.2-6.0 (5.3)*
3.0-5.8 (4.6)*
Ovate
(*) The number in the brackets indicate the average.
31
Palenological Study Table (5): Quantitative and qualitative characters of pollen grains for genus Fritillaria measured by (μm).
No.
Taxa
Polar view (P)
Equaterial view (E)
Exine thickness
Shape Polar view (P)
Shape Equator view (E)
1.
Fritillaria imperialis
53-58 (54.5)*
54-59.8 (57.3)*
1.8
Spherical to semi-spherical
Ovoid to sub-ovoid
2.
F. persica
38-46 (42.4)*
38.6-47 (43.5)*
1.7
Spherical to semi-spherical
Ovoid to sub-ovoid
3.
F. assyariaca
39-47 (43.22)*
40-49 (46.4)*
1.8
Spherical to semi-spherical
Ellipsoid
4.
F. uva-vulpis
42-46 (44.7)*
44-49 (45.4)*
1.8
Spherical to semi-spherical
Ellipsoid
5.
F. crassifolia subsp. kurdika
40-49 (44.9)*
41-49.8 (44.9)*
1.9
Spherical to semi-spherical
Ovoid to sub-ovoid
6.
F. crassifolia subsp. hakkarensis
39-48 (44.7)*
40-49.4 (45.6)*
1.8
Spherical to semi-spherical
Ovoid to sub-ovoid
7.
F. alfreda subsp. glaucoviridis
39-49 (46)*
40-49 (45.7)*
1.8
Spherical to semi-spherical
Ovoid to sub-ovoid
(*) The numbers in the brackets indicate the averages.
37
Ecology & Geographical Distribution Table (8): Distribution of genus Fritillaria of Iraqi Kurdistan region districts, altitudes and types of soil.
No.
(Taxa)
1.
Fritillaria imperialis
2.
F. persica
3.
MJS
MAM
MRO
X
FAR
X
X X
FKI
Type of Soil
088-2220
Rocky clay soil
088-1038
Limestone and clay soil
X
X
F. assyariaca
X
X
X
088-1008
Soil Pocket
4.
F. uva-valpis
X
X
X
028-2188
Damp Soil Pocket
5.
F. crassifolia subsp. kurdica
X
X
1188-2188
Rocky clay soil
X
1088-3008
Lime Stone Soil Cliffs
X
1600-1800
Lime Stone Soil Cliffs
7.
F. crassifolia subsp. hakkarensis F. alfredae subsp. glaucoviridis
X
(*) The numbers in the brackets indicate the averages.
68
X
Altitude(m)
X
6.
X
MSU
Ecology & Geographical Distribution
Months Jan. Feb.
Mar.
Apr.
May.
Species Fritillaria imperialis F. persica F. assyriaca F. uva-vulpis F. crassifolia subsp. kurdiaa F. crassifolia subsp. hakkarensis F. alfredae subsp. glaucoviridis Figure (22): Flowering period for different species of Fritillaria.
77
Jun.
Jul. Aug.
Sep. Oct. Nov.
Dec.
Ecology & Geographical Distribution
Months Jan.
Feb.
Mar.
Apr.
May.
Species Fritillaria imperialis F. persica F. assyriaca F. uva-vulpis F. crassifolia subsp. kurdiaa F. crassifolia subsp. hakkarensis F. alfredae subsp. glaucoviridis Figure (23): Fruiting period for different species of Fritillaria.
78
Jun.
Jul.
Aug.
Sep.
Oct.
Nov.
Dec.
CHAPTER ONE
INTRODUCTION AND LITERATURE REVIEW 1.1 General introduction: Plants are the main sources of food for the human and forage for the animals, in addition to beautifying the nature by their colours and smiles, which make the human to understand the plants and identify the useful one from the harmful and nominate them according to their shapes, tastes or their other distinguished characters which are considered as the beginning of Folk Taxonomy (Al-Ubaidy, 2005). This requirement was related with the human desire to sort the surrounding materials, nominating them and explaining their diversity; such triles are known by Plant Taxonomy (Systematic) (Pandy, 2005 and Potter et al., 2007), after that this science became more accurate in using the invisible characters of the plant and the researchers began moving to the other sciences such as cytology, anatomy, ecology, phytogeography and other biological sciences and even non-biological sciences such as geology and chemistry, (Nasir-Alla, 2007). Taxonomy is regarded as a principle for the other sciences and depending on them as has advantages for many researches and in different fields, and principles come from depending on the information getting from other related scientific fields so it could not be isolated from the other sciences (AbdulMohsin, 2012). Moreover, the new technologies and instruments such as Scanning Electron Microscope (SEM) and Transmission Electron Microscope (TEM) have attributed in developing the taxonomic studies (Abdul-Razaq, 2008 and Mohamad, 2010). Through the importance of taxonomic studies that help in identifying the species numbers of the wild and cultivated plants, especially the economic ones which are a part of the natural wealth of our country, so the 1
CHAPTER TWO
MORPHOLOGICAL STUDY 2.1 Introduction: The study of phenotypic traits of plants are considered as the base foundation to describe the groups of plants and its diagnosis, despite the emergence of recent studies and the development of scientific methods that gave a broad prospect in taxonomic studies, these retained qualities are the lead in making the taxonomist be able to recognize, which is the centerpiece of any advanced taxonomic study and it cannot be dispensed, particularly in the field when the initial diagnosis of the sample and in the development of key taxonomic hierarchy to isolate each other. Through identifying minute phenotypic qualities that became necessary in the field of classification, especially after the discovery of a scanning electron microscope (SEM) and techniques that have helped to solve a lot of problems of taxonomic for being illustrating wider manifestations of the plant, especially when studying the outer appearance and patterning surface of pollen, which gives this type of study the nature of totalitarianism and accuracy in the field of advanced taxonomic studies and then the seed can easily be distinguished the species of it be identified (Diez, 1983 and Abdul-Razaq, 2008). The current study examined phenotypic characteristics of all the vegetative and reproductive organs in detail and according to what is available from scientific means possible to take advantage of them in this aspect. The study was based on what was stated in terms of each of (Lawrence, 1951; Stearn, 1973; Radford et al., 1974 ; Heywood, 1967 ; Al-Musawi, 1987 ; Al-Katib ; 1988 ; Weberling, 1989 ; Christensen , 1992 ; Harlow and Harlow, 1996 ; Aldasoro et al., 1998 and Pandey, 2005).
12
CHAPTER THREE
PALYNOLOGICAL STUDY 3.1 Introduction: The knowledge of pollen morphology can be used as an instrument of multiple scientific researches in Systematic Botany, Paleobotany, Paleoecology, pollen analysis, Aeropalynology, Criminology, Allergy, Stratigraphic correlation of oil-bearing rocks and coal field, drugs in the field of medico palynology and improvement of honey in the field of mellittopalynology and copropalynology, (Akbar and Mir, 2003 and Muhammed, 2010). Pollen morphology provides significant evidences to support the separation of the taxa at different levels of hierarchy. The palynology information has contributed to solve many complex problems facing the investigators in their taxonomic studies. Al-Maadhidy (2003) and Abdul-Razaq (2008) found that the study of pollen is equivalent in its importance as the morphological characters of vegetative and reproductive plant organs. Sharma and Sharma (1972) reported that pollen taxonomy is one of the important factors in identifying and linking the evolutionary and natural relationships between different genera and families. The classification of Lindley (1830) was the first who’s successfully used the pollen studying in isolating the genera of family Orchidaceae. The researches of Fritszche (1832) and Mohl (1835) contributed in substantial coverage of the information in the field of morphology of the pollen. The taxonomic value of the pollen is appeared are numerous and important characters, such as the size of pollen grain, apertures or pores, furrows, their numbers in one grain and the type of ornamentation on the surface of grain (Diez, 1983 and Al-Mashhadani, 2000).
34
CHAPTER FOUR
ANATOMICAL STUDY 4.1 Introduction: There is a remarkable revolution in the investigation of vascular plant anatomy and its use in classification in the last 60 years, the anatomical characters are as valuable as morphological ones and must not be neglected. In taxonomy, every anatomical aspect of plant has been studied without insisting on some and ignoring others, therefore, the quantity of information accumulated are enormous (Stace, 1984). The characteristics of anatomy are of a great importance in modern taxonomic studies and support in many cases, and phenotypic characteristics were interested to many taxonomists, such studies are expanded for the purpose of finding more of the qualities that will help them in isolating species and genera and even families. This is what led to the adoption of the possibility of anatomical traits that comes in the field of diagnostic taxonomic studies, also have applications in the study of phylogenetic relationships (Radford et al. , 1974). The taxonomist’s dependence was increased to use such characters to solve many complex taxonomic problems on the levels of major and minor taxonomic categories, especially after the evolution that happened in the half of last century till now in the techniques of vascular plant anatomy (Metcalfe and Chalk, 1950). Furthermore Davis and Heywood (1973) referred to the importance in identification and study of phylogenetic trends and relationship grades among the taxonomic categories. According to Stace (1984), the characteristics of anatomical value are in parallel to the qualities of phenotypic or less or more of them, to identify 41
CHAPTER FIVE
Molecular Study 5.1 Introduction: Bulb of Fritillaria cirrhosa is an important traditional Chinese herbal medicine. According to the (Chinese Pharmacopoeia, 1995), it is commonly used as an antitussive and expectorant. Many young bulbs from species of Fritillaria are similar to those of F. cirrhosa, but they are different in price and quality. Therefore, there are many young bulbs from species of Fritillaria that could fake those of F. cirrhosa on the commercial market. The coding region of 5S-rRNA is highly conserved in higher eukaryotes. The 5S-rRNA spacer region sequences of F. thunbergii, F. pallidiflora, F. ussuriensi, F. delavayi, F. cirrhosa, F. anhuiensis and F. puqiensis were cloned by PCR with a pair of primers located within the conserved coding region. Based on sequences analyses of the 5S-rRNA spacer region from the 7 species, a specific sequence was found in F. cirrhosa. A pair of specific primers was designed for differentiating the bulbs of F. cirrhosa from each other by PCR (Li et al., 2003). Genetic diversity is critical for adaptation to environmental changes and for the long-term survival of the species. Knowledge of genetic diversity within and among populations has practical importance for conservation and management policies (Hamrick and Godt, 1996 and Fritsch and Rieseberg, 1996). The preservation of genetic diversity within the species is a major target of conservation, because loss of genetic variation is thought to reduce the ability of populations to adapt to environmental change for survival (Hogbin and Peakall, 1999; Honjo et al. , 2004 and Yamagishi et al. , 2010). Therefore, population genetic studies of F. imperialis are essential for providing information needed for conservation of this species. PCR-based DNA fingerprinting techniques such as random amplified polymorphic DNA analysis (RAPD), inter simple sequence 53
CHAPTER SIX
ECOLOGY AND GEOGRAPHICAL DISTRIBUTION 6.1 Introduction: The ecological factors and differences in the geographical distribution have evident effects on the morphological and even anatomical characteristics of the plant and how genes action with environmental factors acts in giving the phenotype characteristics of the individual plant. Environmental factors have played an important role in determining the prevalence and distribution of different species, whether at the level of the world or at the level of different countries (Al-Hili and Al-Ani, 1989 and Al-Mashhadani, 1992). Radford et al. (1974) has confirmed that the researcher has to observe how the morphological characters that are associated with environmental factors to understand the prevalence and population variations depending on soil factors, relative humidity, altitude above sea level and light intensity which affect the plant morphology in many cases, where the altitude and the type of soil and rocks give the evidence of the existence of a certain species (Al-Mashhadani, 1992 and Aldasoro et al. , 2005). We noted that one species shows the variations in the various non-qualitative characters differing in growth locations. The environmental studies, geography and the distribution of species play an important role in estimating and identifying the taxonomic value, which refers to the existence of relationships among the species and also supporting the evolutionary evidences which explain the origin and distribution of species, (Davis and Heywood, 1973; Al-Maadhidy, 2003 and Aldasoro et al. ,2005). Environment is the base of plant taxonomy in understanding and distribution of taxa and composition of flora and understanding the evolutionary relationships of the taxa and variations in population variations and evolutionary adaptations (Al-Mashhadani, 1992). 64
CHAPTER SEVEN
GENERAL DISCUSSION General Discussion: The current study has referred to numeral sides including the study of morphological, palynological, anatomical and molecular characters for the species of the genus Fritillaria grown in Iraqi Kurdistan, in addition to study of environment and distribution of the species in different districts of the country, as well as their taxonomic treatments in detail. Different keys were set for separating the species of genus Fritillaria such as, key of (Zohary, 1946) who mentioned 5 species of this genus within the family Liliaceae that are F. imperialis, F. persica, F. assyariaca, F. kurdica and F. crassifolia in all Iraqi districts. While (Townsend and Geust, 1985) divided the genus in (Flora of Iraq) into 6 taxa that are F.imperialis, F. persica, F. assyariaca, F. uva-vulpis, F. crassifolia subsp. kurdica and F. crassifolia subsp. hakkarensis. Flora of Europe by (Tutin et al. , 1980) separated 23 species of the genus Fritillaria including F. persica. Davis (1984) in (Flora of Turkey) divided this genus into 31 species including F. imperialis, F. persica, F. uvavulpis, F. crassifolia subsp. kurdica, F. crassifolia subsp. hakkarensis and F. assyariaca. Morever, Ridda, T. J. and Dawd, W. H. (1982) divided the genus Fritillaria in to 5 species that are F.imperialis, F. persica, F. assyariaca, F. uvavulpis and F. crassifolia. Parsa (1950) in the key of (Flora Iranica) separated 19 species belong to the genus Fritillaria including the species F. crassifolia, F. kurdica and F. imperialis. Sharma (1998) also separated 2 species of the genus under study that are F.imperialis and F. roylei in the key of (Flora of Upper Liddar Valleys of Kashmir Himalaya). The current study has confirmed that the plants of all the species of the genus 79
80
CHAPTER EIGHT
TAXONOMICAL TREATMENTS Key to the Taxa of the genus Fritillaria: 1. Flowers umblellate, drooping below adense verticillate leaves; tepals more than 5 cm long; perigone red, orange or yellow ……………… 1- F. imperialis 1. Flower solitary to racemose, tepals much shorter, perigone purple or yellow………………………………………………………………….….2 2. Inflorescence a many-flowered ebracteate raceme ……...….. 2- F. persica 2. Inflorescence 2-3 flowered solitary………………3 3. Leaves usually 5-6 per stem, glaucous, linear, stem often papilose at ground level …………………………………. …………………3- F. assyariaca 3. Leaves usually 4 per stem, green, lanceolate, flat; stem smooth ………….4 4. Tepals outer and inner tessellated on a greenish or yellow ground with green strips, inner segments obtuse
…........ 4- F. crassifolia subsp .kurdica
4. Tepal on the outer surface greenish, brownish or much reddish tessellated with broad greenish-yellow stripes tessellated inside, inner segments acute……5 5. Nectar lenghth is (10.0) mm as average with green usually with a black spot at the base colour, filament is papillate…….5- F. crassifolia subsp. hakkarensis 5. Nectar lenghth is (2.5) mm as average with black colour, filaments is glabrous……………………………………………………………………….6 6. Style is filiform and filaments is filliform …………………..6- F. uva-vulpis 6. Style is densely papillose and filaments are stout, papillate bove...................7F. alfredae subsp. glaucoviridis
86
87
CONCLUSIONS AND RECOMMENDATIONS A. Conclusions: A botanical taxonomy of genus Fritillaria has been presented in this thesis. Six species were seen in Iraqi Kurdistan region districts which were Fritillaria imperialis, F. persica, F. assyariaca, F. uva-vulpis, F. crassifolia subsp. kurdica and F.
crassifolia subsp. hakkarensis. Iraqi National
herbarium has already identified four species in the present study which are Fritillaria imperialis, F. persica, F. assyariaca and F. uva-vulpis. The identification of the F. crassifolia subsp. kurdica and F. crassifolia subsp. hakkarensis has been done depending on the information and details of Flora of Iraq, Iran, Turkey and Europe. As a result of the study one sub species were recorded to the first time in Iraq which is F. alfredae subsp. glaucoviridis.
B. Recommendations: 1. Study on elongation of flowering age and period. 2. Using their flowers as priming flowers. 3. Studying the genus in all areas of Kurdistan in a tem in the future, where there are more opportunities to obtain additional information about different characteristics of the including taxa and in more habitats. 4. Study chemotaxonomy.
99
REFFERENCES
Abd-Mohsen, A. S. (2012). A Comparative Systematic Study Of the Genus Allium L. (Alliaceae) in Iraq. College of Education Ibn Al-Haitham, University of Baghdad. Abdul-Razaq, R. T. (2008). A Comparative Systematic Study of Taxa of Subfamily
Pomoideae
(Rosaceae)
in
Iraqi
Kurdistan.
Ph.D.
Dissertation. College of Agriculture, Horticulture, University of Sulaimani. Afroz, S. and Hassan, M. A. (2008). Systematics studies in the family Liliaceae from Bangladish. J. Plant Taxon. 15(2): 115pp. Agostini, G., Echeverrigaray, S., Souza-Chies, T. T., 2008. Genetic relationships among South American species of Cunila D. Royen ex L. based on ISSR. Plant Syst. Evol. 274, 135–141. Akbar A. M. and Mir A. K.. (2003). Pollen Morphology of
Achillea
(Compositae-Anthemoideae) species from Pakistan Pak. J. Weed. Sci. Res. 9 (3 & 4): 253-258, 2003. Al-Ani, B. O. and Q. N. Salih (1988). Principles of Plant Anatomy. High Education Printer. University of Salahadeen. College of Sciences. Erbil. Aldasoro, J. J. and Carlos Aedo (1998). The genus Sorbus (Maloideae, Rosaceae ) in Europe and in north Africa: Morphological analysis and systematics, Botany 23(2).p. 159-212. Aldasoro, J. J., Aedo, C. and Navvaro, C. (2005). Phylogenetic and Phytogeographical relationships in Maloideae (Rosaceae) based on morphological and anatomical characters, Real jardibotanico, consejo, superior, Madrid, Spain. 50: 1.3-32. Al-Hilli, M. R. and H. A. Al-Ani (1989). Plant Ecology. Ministry of High Education and Science Research. Baghdad University. Bait Al-Hikma for publishing, Translating and Distribution. 011
Al-Katib, Y. M. (1988). Taxonomy of Seed Plants. Univ. of Baghdad . (In Arabic). 589pp. Al-Khazraji, T. O. and Falah M. A. (1989). Practical in Plant Anatomy and Microscopic Preparation. Ministry of High Education and Scientific Research University of Salahadeen. P: 321. Al-Maadhidy, A. M. (2003). Classification study for comparing type of species Prunus L. (Rosaceae) in Iraq; Ph.D. Dissertation, College of Agriculture- Musil University. Al-Mashhadani, A. N. (1992). Acomparative systematic Study of Onosma L, (Boraginaceae) in Iraq. ph. D. desertation, College of scinces. Baghdad University. (in Arabic).In Abdul-Razaq , R.T.(2008) A comparative
systematic
study
taxa
of
subfamily
pomoideae
(Rosaceae) in Iraqi Kurdistan. ph. D. desertation. College of Agriculture, Horticulture, University of Sulaimanyaia. Al-Mashhadani, A. N.; and A. H. Al-Musawi. (2000). Morphological chareacterazation of pollen of type of species of Onosma L. in Iraq, Department of Biology, college of Education- Ibin haitham, Baghdad university, jornal of ibn –haitham of Abstract science .vol.13. No.1 Al-Mashhadani, U. N. S. , A. H. Al-Musawi and A. M. Al-Zubaidi (2001). Comparative Anatomical Study of Eight species of the genus Salvia L. (Family Labiatae) in Iraqi. Biology, Department. Ebin Haitham Magazin for the Basic and Practical Sciences. Chapter 15, No. 2. Al-Mayah, A. A. (1983). Taxonomy of Terminalia (Combretaceae), Ph.D. Thesis, Univ. of Leicester. Al-Musawi, A. H. (1987). Plant Taxonomy. University of Baghdad (in Arabic) 379 pp. Al-Rawi, A. (1964). Wild plant of Iraq. Ministry of Agriculture and Irrigation, 3d Edition, Baghdad-Iraq. Pp 18.
010
Al-Rawi, A. (1964). Wild Plants of Iraq with their distribution, Tech . Bull. 14, Dir Gene of Agr. Res. Proj. Ministry of Agriculture, Government Press, P.131. Al-Rawi, A. (1966). Poisonous Plants of Iraq. Ministry of Agriculture and Irrigation, 3d Edition, Baghdad-Iraq. Pp 133 . Al-Rawi, A. (1987). Flora of Kuait. Vol. 2. Alden Press, Oxford: 277-284pp. Al-Rawi, A. and Chakravarty, H. L.
(1964) Medicinal Plants of Iraq.
Ministry of Agriculture and Irrigation, 3d Edition, Baghdad-Iraq. P. 46. Al-Sahhar, Q. F. (1991). Introduction of Plant Classification 2nd Edition. AlDar-Al-Arabia Publisher and Distributer. College of Agriculture University of Cairo. Al-Sawah, D. A. M. (1992). The genus Achillea L. in Iraq. Biosystematic aspects. Ph. D. Thesis, University of Baghdad. (in Arabic, umpublished). Al-Ubaidi, A. K. K. (2005).Systematic study ofthe species of genus Trigonella L. in Iraq. Thesis, Ms.c, Education college, Diala University. P: 198. Archak, S., Gaikwad, A.B., Gautam, D., Rao, E.V.V.B., Swamy, K.R.M., Karihaloo, J.L., 2003. Comparative assessment of DNA fingerprinting techniques (RAPD, ISSR and AFLP) for genetic analysis of cashew (Anacardium occidentale L.) accessions of India. Genome 46, 362– 369. Barkuda, Y.. (2006). The seeds residues. Damascus University Publishes. College of Sciences. P: 191. Bensky, D and Gamble A. (1993). Chinese Herbal Medicine Materia Medica, Eastland Press, Seattle, WA. Budak, H., Shearman, R.C., Parmaksiz, I., Dweikat, I., (2004). Comparative analysis of seeded and vegetative biotype buffalo grasses based on phylogenetic relationship using ISSRs, SSRs, RAPDs, and SRAPs. Theor. Appl. Genet. 109, 280–288. 011
Cao, X. W.; Chen, S. B.; Li, J.; Xiao, P. G. and Chen, S. L. (2008). Steroidal alkaloids from the bulbs of Fritillaria delavayi Franch (Liliaceae). Biochem. Syst. Ecol. 36, 665–668. Chakravarty, H. L. (1976). Plant wealth of Iraq. Vol. 1. Bot. Dir. Agr. Baghdad. 506 Pp. Chong-Zhi Wang1, 2; Ping Li1; Jia-Yi Ding1; Guo-Qian Jin1and ChunSu Yuan2. (2005). Identification of Fritillaria pallidiflora Using Diagnostic PCR and PCR-RFLP Based on Nuclear Ribosomal DNA Internal Transcribed Spacer Sequences. China Pharmaceutical University. Christensen, K. I. (1992). Revision of Crataegus Sect. crataegus and notrosect. Crataeguineae (Rosaceae - Maloideae) in the old world. Systematic Botany monographs 35: 1-199. Davis, P. H. (1984). Flora of Turkey and the East Agean Islands Edinburgh, Univ. Press. 8: 284-303. Davis, P.H. andV. H. Heywood (1973). Principles of Angiosperm taxonomy. Robert,E. Krieger Publishing Company, Huntington, New York. 558 pp. Diez, M. J. (1983). Pollen morphology of genus Anchusa L. (Boraginaceae) and its Taxonomic interest. Pollen Spore 25(3-4)367-382 Dong, Z. and Yu S., (1990). Modern Study and Application of Materia Medica, China Ocean Press, Hong Kong. Ebrahimie, E.; Habashi, A. A.;Mohammadie-Dehcheshmeh, M.; Ghannadha, M.; Ghareyazie, B. and Yazdi-Amadi, B. (2006). Direct shoot regeneration from mature embryo as a rapid and genotype independent pathway in tissue culture of heterogeneous diverse sets of cumin (Cuminum cyminum L.) genotypes. In Vitro Cell Dev. Biol. Plant 42, 455–460. Erdtman G (1969). Handbook of Palynology: Morphology – TaxonomyEcology, An Introduction to the Study of Pollen Grains and Spores. Copenhagen: Munksgaard. 013
Erdtman, G. (1952). Pollen Morphology and Plant Taxonomy, Angiosperms. An introduction to palynology. Almqvist and Wiksell, Stockholm. 530pp. Erdtman, G. (1971). Pollen morphology and plant taxonomy, Angiosperms. European. Volume 2 Rosaceae to Umbelliferae, Cambridge University Press. Vol 2: 469 pp.U.K. Franchi, G. G. and Pacini, E. (2000). Types of pollen dispersal units in Monocots: Wilson, K. L. &Morrison,D. A.(ed.). Monocots-systematics & Evolution. National library of Australia, Sydney: 728 pp. Fritsch, P.W. and Rieseberg, L.H., (1996). The use of random amplified polymorphic DNA (RAPD) in conservation genetics. In: Smith, T.,Wayne, B. (Eds.), Molecular Genetic Approaches in Conservation. Oxford Univ Press, New York, pp. 54–73. Fritzsche, J. (1832). Beitrage Zur kenntnissdes Pollens. Berlin Stettin und Elbing. cited by(Erdtman. G.,1945). Gao, S. L., Zhu, D. N., Cai, Z. H., Jiang, Y. and Zu, D. R. (1999). Organ culture of a precious Chinese medicinal plant- Fritillaria unibracteata. Plant Cell. Tiss. Org. 59, 197–201. Gerrard, J. (1597). The Herball, or Generall Historie of Plantes , 123 p. Gledhill, D. (2008). The names of plants. Cambridge University press, New York: 43p. Guest, E. (1933). Notes on plant and plant products with their colloquial names in Iraq. The Government Press. 27, Baghdad, 111 pp. Guest, E. (1966). Flora of Iraq. Ministry of Agriculture. Republic of Iraq. Vol. 1: 213 pp. Hamrick, J. L. and Godt, M. J.W. (1996). Conservation genetics of endemic plant species. In: Avise, J.C., Hamrick, J.L. (Eds.), Conservation Genetics: Case Histories from Nature. Chapman and Hall, New York, USA, pp. 281–304. 014
Handel-Mazzetti, H. V. (1910). Die veg Lationsverh-hultnisse von Mesopotmies Kurdistan. P. 33. Harlow,W.M., and E .S .Harrar. (1996). Textbook of Dendrology, 8th ed.Mc Graw Hill,Co.New York, 520 pp . Heywood,V. H . (1967). Plant taxonomy. 2nd. ed. Flowering plants of the world. Oxford Univ.Press353 pp. U.K. Hickman (1933). Checklist of vascular plants of san Diego count 4th edition . Hogbin, P.M. and Peakall, R. (1999). Evaluation of the contribution of genetic research to the management of the endangered plant Zieria prostrata. Conserv. Biol. 13, 514–522. Holmgren, P. K. and W. Keuken. (1964). Index Herbarioum. Part, 1. 5th edt. Oothoek, Schttema and Hdkema, Netherlandes. 397pp. Honjo, M.; Ueno, S.; Tsumura, Y.; Washitani, I. and Ohsawa, R. (2004). Phylogeographic study based on intraspecific sequence variation of chloroplast DNA for the conservation of genetic diversity in the Japanese endangered species Primula sieboldii. Biol. Conserv. 120, 211–220. Hooker, J. D. (1894). Flora of British India. Colonial & Indian government, London: 300pp. Hutchinson, J. (1959). The families of flowering plants. Vol. 2. Clarendon press, Oxford: 642-646p. Janssen, P.; Coopman, R.; Huys, G.; Swings, J.; Bleeker, M. and Vos, P.; Zabeau, M. and Kersters, K.1. (996). Evaluation of the DNA fingerprinting method AFLP as a new tool in bacterial taxonomy. Microbiology 142, 1881–1893. Johnson, A. M. (1977). The taxonomy of the flowering plants. Strauss & cramer Gmbh, Germany: 666-667p. Kaneko, K.; Katsuhara, T.; Kitamura, Y.; Nishizawa, M.; Chen, Y.P. and Hsu, H.Y. (1988). New steroidal alkaloids from the Chinese herb drug ‘Bei-mu’. Chem. Pharm. Bull. 35, 4611–4705. 014
Karl W. K. Tsim. (1993). Fritillaria species identification. Unaited states patent, No. 10 Kesari, V.; Sathyanarayana, V.M.;
Parida, A. and Rangan, L. (2010).
Molecular marker-based characterization in candidate plus trees of Pongamia pinnata, a potential biodiesel legume. AoB Plants, 1–12. plq017. Kitajima, J.; Noda, N.; Ida, Y.; Miyahara, K. and Kawasaki, T., 1981. Steroidal alkaloids of fresh bulbs of Fritillaria thunbergii and of crude drug ‘bai-mo’ prepared therefrom. Heterocycles 15, 791–796. Kosenko, V. N. (1991). Pollen Morphology of genus Fritillaria (Liliaceae) Bot anicheskiy Zhurnal (Leningrad) 76: 1201-1210. Kumar, M. ; Mishra, G.P.; Singh, R.; Kumar, J.; Naik, P.K. and Singh, S.B., (2009). Correspondence of ISSR and RAPD markers for comparative analysis of genetic diversity among different apricot genotypes from cold arid deserts of trans-Himalayas. Physiol. Mol. Biol. Plants 15, 225–236. Laurence, Hill. (2012). Fritillaria a list of published names 1-17p. Lawrence, H. M.
(1951). Taxonomy of vascular plants. The Macmillan
Company. New York.838 pp. Li YF; Li YX; Lin J; Xu Y; Yan F; Tang L and Chen F.( 2003). Identification of bulb from Fritillaria cirrhosa by PCR with specific primers. PubMed - indexed for MEDLINE . Li, X.W.; Dai, Y. and Chen, S. L. (2009). Growth and physiological characteristics of Fritillaria cirrhosa in response to high irradiance and shade in age-related growth phases. Environ. Exp. Bot., 77–83. Lindley, J.
(1830). The genera and species of Orchidaceous plants. London.
Cited by (Wodehouse. R. P., 1935). Linnaeus (1853). Flora of China, chen Sing-chi, 24 : 127-133. Linnaeus (1853). Flora of North of America, Vol. 26 Page 15, 58, 164, 173. 015
Ma, X.; Zhang, X. Q.; Zhou, Y. H.; Bai, S. Q. and Liu, W. ( 2008). Assessing genetic diversity of Elymus sibiricus (Poaceae: Triticeae) populations from Qinghai-Tibet Plateau by ISSR markers. Biochem. Syst. Ecol. 36, 514–522. Mathew, B. (1987) Flowering Bulbs for the Garden. The Royall Botanic Gardens, Kew in association with COLLINGRIDGE. The Hamlyn Publishing Group. P. 76-79 Metacalfe, C. R. and Chalk, L. (1950). Anatomy of the Dicotyledons Leaves, Stem and Wood.in relation to Taxonomy with notes on Economic uses Oxford Clarendon press. vol. 2. 1500pp. Milbourne, D.; Meyer, R.; Bradshaw, J. E.; Baird, E.; Bonar, N.; Provan, J.; Powell, W. and Waugh, R., 1997. Comparison of PCR-based marker systems for the analysis of genetic relationships in cultivated potato. Mol. Breed. 3, 127–136. Mohamad, Sh. M. (2010). A Comparative Systematic Study Of Genus Vicia L. (Family: Papilionaceae) In Iraqi Kurdistan. ph. D. desertation. College of Agriculture, Field crop, University of Sulaimanyaia. Mohammadi-Dehcheshmeh, M.; Khalighi, A.; Naderi, A.; Sardari, M. and Ebrahimie, E. (2008). Petal: a reliable explant for direct bulblet regeneration of endangered wild populations of Fritillaria imperialis L. Acta Physiol. Plant 30, 395–399. Mohl, H. (1835). Surla structure et les formes des grains de pollen. Ann. Sci. Nat. 2, 3: 148-180, 304-346. Muschler, R. (1912). A manual Flora of Egypt. Verlag Von J. Cramer, New York: 205p. Nabelek, F. (1929). Iner Turcico-persicum, part Iv. Dela facule des Science de inniversite Masaryk radigees: 30p. Naik, P. K.; Alam, M. F.; Singh, H.; Goyal, V.; Parida, S.; Kalia, S. and Mohapatra, T. (2010). Assessment of genetic diversity through RAPD, ISSR and AFLP markers in Podophyllum hexandrum: a medicinal 016
herb from the Northwestern Himalayan region. Physiol. Mol. Biol. Plants 16 (2), 135–148. Nair, P. K. K. (1971). Pollen Morphology of Angiosperms. Barnes and Nobile, New York: 63p. Nasir-Alla, I. K. (2007). Comparative systematic study of the Wild species of genus Phlomis L. and Sideritis L. (Family Labiatae). Dissertation. University of Baghdad. Ebin-Haitham Education college. Ozler H. and Pehlivan. S. (2007). Comparison of pollen morphological structires of some taxa belonging to Asparagus L. and Fritillaria L. (Liliaceae) from Turkey. Bangladesh J. Bot. 36(2): 111-120. Pandey, B. P. (2005). A textbook of Botany angiosperms. S. Chand and Company, New Delhi: 358-361pp. Pandey, B. P. (2005). Taxonomy of Angiosperms. Department of Botany .J.B. College Baraut Convener, Board of studies and research degree committee C.C.SUniversity, Meerut. 600 pp. Parsa, A. (1950). Flora de Iran. Ministere de L education: Museum D histoire Naturelle de Teheran: 187-199p. Phillips, R. and M. Rix (1981). BULBS. Over 1000 Full-colour photographs of Bulbs in Flower. The Pan Garden Plants. Series. Pan Books Ltd. Cavaye Place, London SW10 qps. Pınar N. M. and Dönmez O. E. (2000). Pollen morphology of Turkish Iris L. (Iridaceae) with reference to evolutionary trends at the infrageneric level. Israel J Plant Sci 48: 129-141. Post, G. E. (1933). Flora of Syria, Palestine & Sinai. Vol. 2. American press, Beirut: 617p. Potter, D; Eriksson, T. R. C.; Evans, S. O. H.; J. E. E. Smedmark; D. R.; Morgan, M.; Kerr, K. R.; Robertson, M. and Arsenault, T. A. (2007). Phylogeny and Classification of Rosaceae . Pl. Syst. Evol.266: 5-43. Printed in Netherlands. 017
Powell, W.; Morgante, M.; Andre, C.; Hanafey, M.; Vogel, J.; Tingey, S. and Rafalski, A. (1996). The comparison of RFLP, RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis. Mol. Breed. 2, 225– 238. Radford, A. E.; W. C. Dikson; J. R. Massy and C. R. Bell. (1974). Vascular plants systematics. Harber & Row. New York. 891 pp. Ravash, R.; Shiran, B.; Alavi, A. A.; Bayat, F.; Rajaee, S. and Zervakis, G. I. (2010). Genetic variability and phylogenetic relationships of Pleurotus eryngii speciescomplex isolates from Iran based on RAPD and ITS sequences. Mycol. Progr. 9, 181–194. Rechinger, K. H. (1964). Flora of Lownland Iraq. Weinheim Verlag von. J. Cramer New York Hafner Co. 764pp. Rendle, A. B. (1953). The classification of flowering plants. Vol. 1. University press, Cambredge: 412 pp. Ridda, T. J. and Daoud, W. H. (1983). Geographical distribution of Wild vascular plants of Iraq. National herbarium of Iraq, p 106. Rix, E. M. (2001). Fritillaria: A Revised Classification. Edinburgh: The Fritillaria Group of the Alpine Garden Society, United Kingdom. Sass, J. E. (1958). Botanical micro technique. 3rd. Edition the lowa stat University press.228 p. In Abdul-Razaq , R.T.(2008) A comparative systematic study taxa of subfamily pomoideae (Rosaceae) in Iraqi Kurdistan. ph. D. desertation. College of agriculture, horticulture, university of sulaimanyaia. Sehgal, D.; Rajpal, V. R.; Raina, S. N.; Sasanuma, T. and Sasakuma, T. (2009). Assaying polymorphism at DNA level for genetic diversity diagnostics of the safflower (Carthamus tinctorius L.) world germplasm resources. Genetica 135, 457–470. Sharma, A .K., and A.Sharma (1972) .Chromosome techniques. Theory and Practice 2nd.ed.Butterwoths.London.575 pp. 018
Sharma, B. M. (1998). Flora of Upper Liddar Valleys of Kashmir Himalaya. Vol. 2, p. 204. Shiran, B.; Amirbakhtiar, N.; Kiani, S.; Mohammadi, S.; Tabatabaei, B. E. and Mordi, H. (2007). Molecular characterization and genetic relationship among Iranian almond cultivars assessed by RAPD and SSR markers. Sci. Hortic. 111, 280–292. Simpson, M. G. (2006). Plant Systematics. Elsevier Academic press, Amstedam: 180-590 pp. Singh, G. (2004). Plant systematics: an integrated approach. Science Publisher, India: 351pp. Solerder, H. (1908). Systematic anatomy of the dicotyledons. Clarendon Press, Oxford. Vol. 1. 636-642. Stace, C. A. (1984). Plant Taxonomy and Biosystematic Great Britain at the Pitman Press . Bath . 267 pp. Stearn, W. (1973). Botanical Latin. David & Charles. London . 566 pp. Stuessy, T. F. (1990). Plant Taxonomy. The systematic evolution of comparative data Columdia University Press New York. Ta``ckholm, V. and Drar, M. (1954). Flora of Egypt, Vol. 3. Cario University Press, Cairo: 644p. Takhtajan, A. (2009). Flowering Plants. Springer Sciense-Business Media, Russia: 871 pp. Teksen, M.; Z. AYTAC, and Nur, M. P. (2010). Pollen morphology of the genus Fritillaria L. (Liliaceae) in Turkey. Aksaray University, Faculty of Arts and Science, Department of Biology, 397-416pp. Tohme, G. and H. Tohme (2007). Illustrated Flora of Lebanon. 200 wild flowers. National council for Scientific Research. CNRS Publication. 2007. P. 397-398. Townsend, C. C. and Evan Guest. (1985). Flora of Iraq Vol .8, Ministry of Agriculture, Baghdad, 76-83 pp. 001
Tutin, T.G.; D. M. Moore; V. H. Heywood ;N. A. Burges; D.H. Valentine; S.M Walters, and D.A. Webb` ( 1980) . Flora Europaea . Camberidge, Univ. Press , London and New York , Vol. 5.p.31-35. Verma, P. C.; Chakrabarty, D.; Jena, S. N., Mishra, D. K.; Singh, P. K.; Sawant, S. V. and Tuli, R. (2009). The extent of genetic diversity among Vanilla species: comparative results for RAPD and ISSR. Ind. Crops Prod. 29, 581–589. Vos, P.; Hogers, R.; Bleeker, M.; Reijans, M.; van de lee, T.; Hornes, M., Frijters, A.; Pot, J.; Peleman, J.; Kuiper, M. and Zabeau, M.(1995). AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res. 23, 4407–4414. Wang, C. Z.; P. Li; J. Y. Ding; G. Q . Jin and C. S. Yuan. (2005). Identification of Fritillaria pallidiflora using diagnostic PCR and PCR-RFLP based on nuclear ribosomal DNA internal transcribed spacer sequences. China Pharmaceutical University. Wang, S. J.; Yu, J. L.; Gao, W.Y.; Pang, J. P.; Yu, J. G. and Xiao, P. G. (2007). Comparison of starches separated from three different F. cirrhosa. J. Food Eng. 80, 417–422. Weberling, F. (1989). Morphology of flowers and inflorescences. Department of Biology in Ulm .405 P. Weising, K.; Nybom, H.; Wolf, K. and Meyer, W. (1995).DNA Fingerprinting in Plants and Fungi. CRC Press, Boca Raton, Fla., 322 pp. Wendelbo, P. (1985). Allium L. In: Towensend, C.C. and Guest, E.(eds.), Flora of Iraq. Vol. 8, Ministry of Agriculture Republic of Iraq, Baghdad. 440 pp. Williams, J. G. K.; Kubelik, A. R.; Livak, K. J.; Rafalski, J. A. and Tingey, S.V. (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18, 6531–6535. Willlis, J. C. (1955). A dictionary of flowering plants & ferns, ed., 6. Cambridge, University Press: 384p. 000
Wodehouse, R. P. (1935). Pollen grains, their structure identification and significance in science and medicine. Hafner Publishing Company. New York & London. 574 pp. Yamagishi, M.; Nishioka, M. and Kondo, T. (2010). Phenetic diversity in the Fritillaria camschatcensis population growen on the Sapporo campus of Hokkaido University. Landsc. Ecol. Eng. 6, 75–79. Yang Shou-Zhong (translator), (1998). The Divine Farmer's Materia Medica, Blue Poppy Press, Boulder, CO. Yu, H. H.; Yang, Z. L.; Sun, B. and Liu, R. N. (2011). Genetic diversity and relationship of endangered plant Magnolia officinalis (Magnoliaceae) assessed with ISSR polymorphisms. Biochem. Syst. Ecol. 39, 71–78. Yu, R. P.; Z. F. Cheng and X. Q. Gong. (2007). Determination and comparison of trace elements in four sorts of Bulbus Fritillariae by microwave digestion-atomic absorption spectrometry. Spectrosc. Spect. Anal. 27, 2591–2594. Zietkiewicz, E.; Rafalski, A. and Labuda, D. (1994). Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20, 176–183. Zohary, M. (1946). The Flora of Iraq and its Phytogeographycal subdivision. Iraq. Dept. Agr. Bull. 3-Baghdad. 201pp.
001
