Post on 19-Jan-2020
A MORPHOLIGICAL AND MOLECULAR PHYLOGENETIC ANALYSIS OF MALAYSIAN
Kerivoula (Chiroptera: Vespertilionidae)
Noor Haliza binti Hassan @ Ahmad
QL 737 C595 N818 2009
Master of Science 2009
P. KHIDMAT MAKLUMAT AKADEMIK UNIMAI
11111 IIIIINIIIIIIIIN 1000212172 A Morphological and Molecular Phylogenetic Analysis of Malaysian Kerivoula (Chiroptera:
Vespertilionidae)
NOOR HALIZA BINTI HASAN
A thesis submitted in fulfillment of the requirement for the degree of
Master of Science
Faculty of Resource Science and Technology UNIVERSITI MALAYSIA SARAWAK
2009
DECLARATION
I hereby declare that no portion of the work referred to this thesis has been submitted in support
of an application for another degree or qualification to this or any other university or institute of
higher learning.
(Noor Haliza binti Hasan @ Ahmad)
JUNE 2009
1
Acknowledgements
Bismillahhirrahmannirrahim. Alhamdulillah and grace the Allah Almighty to have given me
strength and determination, for have given me the health and faith to finally complete my work
for my master's degree. My deep appreciation goes to my supervisor, Prof. Dr. Mohd Tajuddin
Abdullah (MTA) and co-supervisor, Assoc. Prof. Dr. Edmund Sim for their guidance, advice,
constructive comments, concern and support throughout this study. This study would not have
been possible without various administrative and financial support from UNIMAS. I would like
to thank UNIMAS for granting me the Zamalah UNIMAS 2007/09 scholarship. This study was
also funded by three grants; the Ministry of Science, Technology and Innovation IRPA grant
number 09-02-09-1022-AE001 lead by MTA with co-researchers, Yuzine Esa and Awg Ahmad
Sallehin Awg Husaini; grant FRGS 06(08)6602007(25) and UNIMAS Small Grant Scheme
awarded to MTA. I would also like to thank Sarawak Forestry Corporation and Sarawak
Forestry Department for granting permission with license number 07775 under the State Wild
Life Protection Rules 1998; for research permit number NPW. 907.4.2(II)-54 and permit to enter
park number 45/2007. My thanks also go to the Institute of Biological Diversity (IBD) Krau
Pahang, Department of Wildlife and National Park (DWNP) Kuala Lumpur and Musuem of
Sabah, Kota Kinabalu, Sabah.
I wish to thank members of the Department of Zoology; Mr Besar Ketol, Mr. Isa Sait, Mr. Huzal
Irwan Husin, Mr. Wahab Mami and Mr. Mohd Jalani Mortada for their hard work and
assistances throughout this study. To Molecular Ecology Laboratory (MEL) seniors; Andy Kho
Han Guan, Jayaraj Vijaya Kumaran, Fong Pooi Har and Siti Nurlydia Sazali, I wish to thank
them for being the steady mentors for me since 2005 until the very end of this study. My thanks
It
also go especially for Mr. Faisal Anwarali Khan for sharing his samples, ideas and thoughts
throughout these years, and Ms. Ratnawati Hazali, for being a steady supporter and a big sister
whom I can always turn to for advice. I thank my MEL colleagues; Hung Tze Mau, Roberta
Chaya Tawie Tingga, Eileen Lit, Mohd Ridwan Abd Rahman, Anang Setiawan Achmadi, Sigit
Wiantoro, Muhd Ikhwan Idris, Mohd Fizl Sidq Mohd Ram i, Eric Pui Yong Meng and Nur
Salmizar Azmi for their wonderful friendship, for the great times, shared knowledge and support.
Never forgotten, to my best friends, Shamsyah Hamid, Rohanie Bohan, Ahmad Farizzulkhairi
Ahmad Sobri, Khairul Anwar Othman and to my good friends, Nur Hafizah Azizan, Ida Nivina
Pathe, Aminah Imat and Pang Sing Tyan; thanks for all the wonderful memories, friendship,
support and encouragement.
I would like to thank my beloved parents, Hasan @ Ahmad bin Anis and Jama'yah binti Kedri
for their never ending encouragement, support and prayers. To my siblings, Muhammad Najib
bin Hasan, Muhammad Fakhri bin Hasan, Muhammad Al-Hadi Akmal bin Hasan, Muhammad
Fakhrul Aiman bin Hasan and Muhammad Fakhrul Izzat bin Hasan; always have faith and
always be thankful. And to Omu, thank you for always being there and always being supportive.
111
Abstract
(Morphometric and phylogenetic analyses were done on six species of Kerivoula from Malaysia
(Morphological studies could only be done on five out of the six Kerivoula species available for
this research, namely K. papillosa, K. lenis, K. pellucida, K. hardwickii and K. minutaJ No
sample of K intermedia was available for morphological analysis. (Thirty-one
characters of the
external body, skull and dentition were taken from 47 adult individuals of Kerivoula(Three
separate analyses were done on the morphological data; (1) clustering analysis, (2) principal
component analysis (PCA) and (3) discriminant function analysis (DFA) were applied to the
data)(The findings from all the three analyses supported the groupings of the Kerivoula samples
into six different groups; namely, K minuta, K. hardwickii, K. pellucida, K. lenis and K.
papillos) Cryptic samples of K papillosa were further separated into two types; K. papillosa
type small (K papillosa type S) and K papillosa type large (K papillosa type L). Phylogenetic
analysis was done on six available Kerivoula samples utilising three mitochondrial genes; 409
basepair (bp) of cytochrome b (cyt b), 478 bp of cytochrome oxidase I (COI) and 1044 bp of
NADH dehydrogenase subunit 2 gene (ND2). This was followed by another analysis ustilising
one nuclear gene - 1054 bp of recombinant activating gene subunit 2 (Rag2). The
reconstructions of phylogenetic trees depicting the relationship of Kerivoula were retrieved using
all four inferring methods through three analyses namely, neighbor joining (NJ), maximum
parsimony (MP) and maximum likelihood (ML). All analyses consistently resulted in seven
groups, namely K minuta, K pellucida, K. hardwickii, K. lenis and K. papillosa, with K.
papillosa further separated into two different subgroups which were congruent with those of
morphological analyses, and also the addition of K intermedia samples. The same samples were
used in both analyses and out of 17 samples identified as K papillosa, five were classified as K.
iv
papillosa type L (forearm length of 44.5 mm to 49.0 mm); with one sample from the Madai Cave
in Sabah and the other four from the Niah National Park of Sarawak. Another 12 samples were
classified as K papillosa type S (forearm length of 40.0 mm to 44.5 mm) with their distribution
scattered around Sarawak and Peninsular Malaysia. The separation of these two types of K
papillosa were also supported by a notable genetic distance of >10% which was comparable to
those of biologically different species. This suggested the presence of cryptic species within the
K papillosa groups. It was also noted that K hardwickii samples were separated into the Eastern
and the Western Borneo samples with 100% support of bootstrap for the cyt b and the ND2
genes analyses. The existence of a potential phylogroup was suggested with a genetic distance of
4.6% to 6.0% in the cyt b as >5% was the value proposed by Baker and Bradley (2006) for such
definition. Separation of K minuta samples into two subgroups was also observed using at least
three analyses and the existence of a subspecies was suggested. It was concluded that the
analysis using ND2 gene gave the best tree in depicting the phylogenetic relationship of
Kerivoula. The comparison of the K papillosa type S and the K papillosa type L identified in
this study to the type specimen would justify the taxonomic revision of the cryptic species within
the genus. Population studies of K papillosa, K hardwickii and K minuta were suggested to
further verify the findings of the present study. Further analysis onto both forms of K papillosa
together with the other nine Kerivoula species occurring in Malaysia would provide better
insights into the phylogenetic relationship of genus Kerivoula. (The findings of this study were
expected to aid in the taxonomy and future management and conservation plans for this genus
Keywords: Kerivoula, Malaysia, morphology, molecular phylogenetic, cryptic species
V
Ana/isis morfologi dan filogenetik moleku/ Kerivoula (Chiroptera: Vespertilionidae) darf
Malaysia
Abstrak
Analisis ke alas morfomelrik dan hubungan ftlogenetik telah dijalankan ke alas enam spesies
Kerivoula yang lerdapat di Malaysia. Kajian morfologi hanya dijalankan ke alas lima daripada
enam spesies Kerivoula, iaitu K. papillosa, K. lenis, K. pellucida, K. hardwickii dan K. minula
herikutan liada sampel K. intermedia yang dimiliki semasa kajian dijalankan. Tiga puluh salu
ciri ukuran luar hadan, iengkorak dan gigi telah diambil ke alas seliap 47 individu Kerivoula
yang dewusa dan tiga analisis yang berbeza telah di jalankan ke alas semua data ukuran yang
telah diamhil; (1) "clustering analysis ", (2) analisa "principal component " dan (3) analisa
"discrimirnrnt . /unction". Ketiga-tiga analisis ini telah menyokong pembahagian . sampel
Kerivoula yang ada kepada enam spesies, iailu K. minula, K. hardwickii, K. pellucida, K. lenis
dan K. papillosa. Sampel-sampel K. papillosa yang kriptik kemudiannya leluh dihahagi kepuda
duu jenis, iailu K. papillosa jenis kecil (K. pa illosa jenis S- "small ") dun K. papillosu jenis
bestir (K. papillosa jenis L- "large'). Analisis filogenetik pula telah dijalankan ke alas enam
spesies Kerivoula den gun menggunakan liga gen mitokondria, iuitu 409 pasungan hes dari
jujukun gen sitokrom h (cyt J, 478 pasangan hes dari jujukran gen . sitokrom ok. sidu I(C'C)1) dan
1044 pasangan bes dari jujukan gen NADH "dehydrogenuse " subunit 2 (ND2). Ini diikuli
unulisis yang menggunukun 1054 pasangan bes dari jujukan gen nuklear ukiivusi rekomhinun
subunit 2 (Rag2). Pembinaan semula pokok yang menggambarkan hubungan. filogenetik uniuk
Kerivoula mcný, >gunakan keempal-empal gen terse but lelah dijalankan melalui tiKu unulisis iuilu
"neighbor-joining" (NJ), "maximum parsimony" (MP) dan "maximum likelihood" (Ml, ).
vi
Penambahan sampel dari spesies K intermedia untuk analisis filogenetik ini telah
membahagikan sampel-sampel yang ada kepada tujuh kumpulan, iaitu K. intermedia, K. minuta,
K pellucida, K. hardwickii dan K lenis, dengan K. papillosa yang kemudiannya dibahagikan
lagi kepada dua jenis, sebagaimana yang didapati dari hasil kajian morfologi. Dari 17 sampel
yang dikenalpasti sebagai K. papillosa, lima daripadanya diklasifikasikan sebagai K. papillosa
jenis L (ukuran lengan 44.5-49.0 mm), dengan satu sampel didapati dari Gua Madai di Sabah
dan empat sampel yang lain didapati dari Taman Negara Niah, Sarawak. Dua belas sampel
yang seterusnya diklasifikasikan sebagai K. papillosa jenis S (ukuran lengan 40.0-44.5 mm) dan
didapati dari pelbagai lokasi di sekitar Sarawak danjuga Semenanjung Malaysia. Pembahagian
K. papillosa kepada dua jenis ini juga disokong oleh peratusan jarak genetik >10%, yang mana
bersamaan dengan peratusan di antara spesies yang berbeza dari segi biologi. Ini mungkin
menunjukkan bahawa terdapat kewujudan spesies kriptik di dalam kumpulan K papillosa.
Kajian ini juga telah mendapati bahawa kumpulan K. hardwickii telah dibahagikan kepada
kumpulan Borneo Barat dan Borneo Timur. Ini disokong oleh nilai 'bootstrap' sebanyak 100%
di dalam analisis untuk gen `cyt b' dan juga gen ND2. Perbezaan peratusan jarak genetik di
antara kedua-dua kumpulan ini adalah sebanyak 4.6-6.0%, dan peratusan jarak genetik
sehanyak >5% telah dicadangkan oleh Baker dan Bradley (2006) untuk kemungkinan wujudnya
phylogroup' yang belum dikenalpasti di dalam kumpulan berkenaan. Pembahagian kumpulan
K minuta kepada dua subkumpulan juga telah didapati daripada liga analisis genetik dari
kajian ini. Dengan ini juga dicadangkan bahawa terdapat kewujudan subspesies bagi K. minuta.
Dengan ini, disimpulkan bahawa analisis yang menggunakan gen ND2 memberi gambaran yang
terbaik tentang hubungan filogenetik di antara spesies dalam genus Kerivoula untuk kajian ini.
Perhandingan di antara K. papillosa jenis L dan K. papillosa jenis S terhadap -type specimen "
vii
K. papillosa mungkin akan dapat memberi penjelasan ke Was status sampel ini di dalam genus
Kerivoula. Kajian lanjutan ke atas populasi genetik untuk spesies K. papillosa, K. hardwickii
dan K. minuta adalah dicadangkan bagi memberi gambaran yang lebih jelas terhadap hasil
yang didapati dari kajian ini. Kajian selanjutnya ke atas kedua-dua jenis sampel K papillosa
bersama sembilan lagi spesies Kerivoula yang terdapat di Malaysia mungkin akan memberi
gambaran yang lebih lengkap ke Was hubungan filogenetik di antara spesies di dalam genus
Kerivoula di Malaysia. Hasil kajian ini diharapkan dapat membantu didalam pengurusan dan
pelan-pelan pemeliharaan genus ini di masa hadapan.
Kata kunci: Kerivoula, Malaysia, morfologi, filogenetik molekul, spesies kriptik
Vlll
Pusai Ktuuwii, . v. as,;. tuat ruc. adenilc UNIVERSITI MALAYSIA SARAWAIC
Table of Contents
Declaration
Acknowledgement
Abstract
Abstrak
Table of content
List of Figures
List of Tables
Abbreviations
CHAPTER ONE General Introduction
1.1 Chiroptera (Bats)
1.1.1 Classifications of Chiroptera
(i) Megachiroptera
(ii) Microchiroptera
1.1.2 Taxonomic Studies of Chiroptera
1.1.3 New Classifications of Bats
1.2 Study Taxa: The Wooly Bats (Kerivoula sp. )
1.2.1 New Species in Kerivoula
1.2.2 Distribution and Conservation Status
1
11
iv
V1
ix
xviii
xxiii
xxvi
1
3
4
5
7
9
10
12
ix
1.3 Morphology Analysis
1.3.1 Cluster Analysis
1.3.2 Principal Component Analysis
1.3.3 Discriminant Function Analysis
1.3.4 Other Statistical Analysis
1.4 Molecular Approaches
1.4.1 Mitochondrial DNA
(i) Cytochrome b
(ii) Cytochrome Oxidase I
(iii) NADH Dehydrogenase gene subunit 2
1.4.2 Nuclear Gene
(i) Recombinant Activating Gene 2
1.4.3 Inferring Methods
15
16
18
19
21
23
24
24
25
26
26
27
1.5 Rationale 28
1.6 Aims of Study
1.6.1 Morphological Analysis
1.6.2 Molecular Phylogenetic Analysis
1.6.3 Morphological and Phylogenetic Analysis
32
32
32
32
X
1.7 Outline of thesis
CHAPTER TWO Material and Methodologies
2.1 Specimen Collections
2.1.1 Collecting Methods and Sites of Study
2.1.2 Specimen Identification
2.1.3 Specimen Preservation
2.1.4 Museum Voucher Samples
2.2 Morphological Study Materials
2.2.1 Skull Extraction and Measurements
2.3 Morphological Analysis
2.3.1 Cluster Analysis
2.3.2 Basic Statistical Test
2.3.3 Principal Component Analysis (PCA)
2.3.4 Discriminant Function Analysis (DFA)
2.4 Molecular Study
2.4.1 Sources of DNA
2.4.2 Laboratory Materials
33
34
34
35
35
39
41
41
42
43
43
44
X1
2.5 Molecular Methodologies
2.5.1 DNA Extraction
2.5.2 DNA Visualization
2.5.3 Amplification using Polymerase Chain Reaction
(a) Mitochondrial Gene
(b) Nuclear Gene
2.5.4 Purification and Sequencing
2.6 Molecular Analysis
2.6.1 Distance-based Method
2.6.2 Character-based Methods
CHAPTER THREE Morphology Analysis Results
3.1 Morphological Data and Cluster Analysis
3.2 Statistical Tests
3.2.1 Multiple Regression
3.2.2 Normality Test and Data Transformation
3.2.3 Homogeneity of Variance Test
3.2.4 Multicolinearity Test
3.3 Principal Component Analysis
45
46
47
48
49
50
51
51
52
53
59
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60
60
61
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3.4 Discriminant Function Analysis
CHAPTER FOUR Molecular Analysis Results
4.1 Mitochondrial Gene Analysis
4.1.1 Cytochrome b
4.1.2 NADH Dehydrogenase gene subunit 2
4.1.3 Cytochrome Oxidase I
4.2 Nuclear Gene Analysis
4.2.1 Recombinant Activating Gene subunit 2
CHAPTER FIVE Discussion
5.1 Morphological Analysis of Kerivoula
5.2 Phylogenetic Relationship of Kerivoula
65
69
81
90
99
107
5.2.1 Mitochondrial Gene Analysis 114
5.2.2 Nuclear Gene Analysis in Comparison with the mtDNA Gene 118
Analysis
X111
5.3 Comparison of Morphology and Molecular Results 119
CHAPTER SIX General Conclusions and Recommendations 123
REFERENCES 126
List of Publications 143
APPENDIX A Samples used for morphometrics analysis. 144
APPENDIX B Multiple regression for testing sex dimorphism for each 152
species.
APPENDIX Cl P-values of Kolmogorov-Smirnov goodness-of-fit test for 153
all characters in K papillosa type S.
APPENDIX C2 P-values of Kolmogorov-Smirnov goodness-of-fit test for 154
all characters in K papillosa type L.
APPENDIX C3 P-values of Kolmogorov-Smirnov goodness-of-fit test for 155
all characters in K. pellucida.
xiv
APPENDIX C4 P-values of Kolmogorov-Smirnov goodness-of-fit test for 156
all characters in K hardwickii.
APPENDIX C5 P-values of Kolmogorov-Smirnov goodness-of-fit test for 157
all characters in K minuta.
APPENDIX D Box's M test results for standardised characters. 158
APPENDIX E Variables in the analysis and not in the analysis for 159
normally distributed character and all characters.
APPENDIX F1 Scree plot describing the eigenvalue for each component 165
in the analysis.
APPENDIX F2 Total variance explained by each factor. Extracted factor 166
used in analysis are being bold.
APPENDIX F3 Communalities tables for the extracted component in the 167
analysis.
APPENDIX F4 KMO and Bartlett's test. 168
xv
APPENDIX F5 Component score coeffiecient matrix for all 31 variables 168
used in the analysis.
APPENDIX F6 Descriptive statistics for the variables used in the analysis. 169
APPENDIX G1 Classification results for all characters in analysis. 170
APPENDIX G2 Statistical test used at each step of the stepwise analysis 172
(Wilk's lambda) for all characters.
APPENDIX G3 Wilk's Lambda test of functions and eigenvalues for all 173
functions.
APPENDIX H The aligned sequences of the 409 bp of partial cyt b 174
sequences for each species used in Chapter Four.
APPENDIX I The aligned sequences of the complete ND2 sequences for 180
each species used in Chapter Four.
APPENDIX J The aligned sequences of the partial COI sequences for 191
each species used in Chapter Four.
xvi
APPENDIX K The aligned sequences of the partial Rag2 sequences for 196
each species used in Chapter Four.
APPENDIX L Comparison of skulls of K papillosa type L, K papillosa 204
type S, K lenis, K pellucida and K hardwickii.
xvii
List of Figures
Figure 2.1 Sampling sites for Kerivoula samples used in present study, namely as
the following; 1-Kubah National Park, Swk; 2-Batang Ai National Park,
Swk; 3-Bau, Swk; 4-Bintulu, Swk; 4-Niah National Park, Swk; 5-
Loagan Bunut National Park, Swk; 6- Lambir Hills National Park, Swk;
7-Similajau National Park, Swk; 8-Gua Madai, Sbh; 9-Taman Negara
Pahang, PM; 10-Kelantan, PM; 11-Perak, PM; 12-Johor, PM and 13-
Terengganu, PM. *Swk=Sarawak; Sbh=Sabah and PM=Peninsular
Malaysia.
Figure 2.2 Thirty-one characters used for measurements and analysis in Kerivoula.
Drawing is not to scale.
Figure 3.1 Figure above showed the dendogram for cluster analysis result using
UPGMA analysis and average Euclidean distance method. K papillosa
S1=Sarawak (SWK) locality; K papillosa S2=Peninsular Malaysia
(PM) locality; K pellucida 1=SWK locality; K pellucida 2=PM
locality; K minuta 1=SWK locality; K minuta 2=PM locality;
FA=forearm length; mm=millimeter.
Figure 3.2 Regression factor plot for species grouping for Factor 1 versus Factor 2.
XVlil
Figure 3.3 Regression factor plot for species grouping for Factor 1 versus Factor 3.
Figure 3.4 Canonical variate analysis for all characters using pooled covariance
matrix.
Figure 3.5 Canonical variate analysis for all characters using separate covariance
matrix.
Figure 4.1 Plot of transition and transversion against divergence using Kimura
(1980) distance method onto the third codon position shows no
saturation occurrence in cyt b gene region.
Figure 4.2 Phylogenetic relationships of six species of Kerivoula under study based
on 409 bp partial cyt b mtDNA gene sequences. The phylogeny is a
single tree recovered using NJ analysis. Values on the branches
represent NJ bootstrap estimates, based on 1000 replicates. Only
bootstrap values >50% are shown.
Figure 4.3 Unweighted and rooted MP tree based on nucleotide data set of 409 bp
partial cyt b mtDNA gene (tree length=440; CI=0.4295; RI=0.7794).
Values on the branches represent MP bootstrap estimates, based on
1000 replicates. Only bootstrap values >50% are shown.
xix
Figure 4.4 Rooted ML tree (-Ln likelihood= 2652.61175) generated based on
nucleotide data set of 409 bp partial cyt b mtDNA gene. Values on the
branches represent ML bootstrap estimates, based on 100 replicates.
Only bootstrap values >50% are shown.
Figure 4.5 Plot of transition and transversion against divergence using Kimura
(1980) distance method onto the third codon position shows no
saturation occurrence of the ND2 gene region.
Figure 4.6 Phylogenetic relationships of six species of Kerivoula under study based
on 1044 bp complete ND2 mtDNA gene sequences. The phylogeny is a
single tree recovered using NJ analysis. Values on the branches
represent NJ bootstrap estimates, based on 1000 replicates. Only
bootstrap values >50% are shown.
Figure 4.7 Unweighted and rooted MP tree based on nucleotide data set of 1044 bp
complete ND2 mtDNA gene (tree length=1256; CI=0.5645;
RI=0.7760). Values on the branches represent MP bootstrap estimates,
based on 1000 replicates. Only bootstrap values >50% are shown.
xx
Figure 4.8 Rooted ML tree (-Ln likelihood= 7256.95353) generated based on
nucleotide data set of 409 bp partial cyt b mtDNA gene. Values on the
branches represent ML bootstrap estimates, based on 100 replicates.
Only bootstrap values >50% are shown.
Figure 4.9 Plot of transition and transversion against divergence using Kimura
(1980) distance method onto the third codon position shows no
saturation occurrence of COI gene region.
Figure 4.10 Phylogenetic relationships of six species of Kerivoula under study based
on 478 bp partial COI mtDNA gene sequences. The phylogeny is a
single tree recovered using NJ analysis. Values on the branches
represent NJ bootstrap estimates, based on 1000 replicates. Only
bootstrap values >50% are shown.
Figure 4.11 Unweighted and rooted MP tree based on nucleotide data set of 478 bp
partial COI mtDNA gene (tree length=339; CI=0.6077; RI=0.8222).
Values on the branches represent MP bootstrap estimates, based on
1000 replicates. Only bootstrap values >50% are shown.
Figure 4.12 Rooted ML tree (-Ln likelihood= 2250.21816) generated based on
nucleotide data set of 478 bp partial COI mtDNA gene. Values on the
branches represent ML bootstrap estimates, based on 100 replicates.
xxi
Only bootstrap values >50% are shown.
Figure 4.13 Plot of transition and transversion against divergence using Kimura
(1980) distance method shows no saturation occurrence in the Rag2
gene region.
Figure 4.14 Phylogenetic relationships of six species of Kerivoula under study based
on 1054 bp partial Rag2 nuclear gene sequences. The phylogeny is a
single tree recovered using NJ analysis. Values on the branches
represent NJ bootstrap estimates, based on 1000 replicates. Only
bootstrap values >50% are shown.
Figure 4.15 Unweighted and rooted MP tree based on nucleotide data set of 1054 bp
partial Rag2 nuclear gene (tree length=118; CI=0.7542; RI=0.8564).
Values on the branches represent MP bootstrap estimates, based on
1000 replicates. Only bootstrap values >50% are shown.
Figure 4.16 Rooted ML tree (-Ln likelihood= 2228.30394) generated based on
nucleotide data set of 1054 bp partial Rag2 nuclear gene. Values on the
branches represent ML bootstrap estimates, based on 100 replicates.
Only bootstrap values >50% are shown.
XXII