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Sperata Sarwari) in the PUNJAB, PAKISTAN

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Sperata Sarwari) in the PUNJAB, PAKISTAN GENETIC ANALYSIS IN DIFFERENT RIVERINE POPULATION OF CARNIVOROUS FISH SPECIES (Sperata sarwari) IN THE PUNJAB, PAKISTAN By Asma Aziz 2013-GCUF-0502-4 Thesis submitted in partial fulfilment of the requirements for the degree of DOCTORATE OF PHILOSOPHY IN ZOOLOGY DEPARTMENT OF ZOOLOGY GOVERNMENT COLLEGE UNIVERSITY, FAISALABAD. September, 2017 i DEDICATION Dedicated to my Affectionate Parents, And My Amicable Teachers ii DECLARATION The work reported in this thesis was carried out by me under the supervision of Prof. Dr. Farhat Jabeen, Chairperson, Department of Zoology, Govt. College University, Faisalabad, Pakistan. I hereby declare that the title of thesis “Genetic analysis in different riverine population of carnivorous fish species (Sperata sarwari) in the Punjab, Pakistan” and the contents of thesis are the product of my own research and no part has been copied from any published source (except the references, standard mathematical or generic models /equations /formulas/protocols etc.). I further declare that this work has not been submitted for the award of any other degree /diploma. The University may take action if the information provided is found inaccurate at any stage. Signature of the Student/Scholar Name of Student: Asma Aziz Registration No: 2013-GCUF-0502-4 iii Plagiarism Undertaking I solemnly declared that research work presented in the thesis titled “Genetic analysis in different riverine population of carnivorous fish species (Sperata sarwari) in the Punjab, Pakistan” is solely my research work with no significant contribution from any other person. Small contribution/help wherever taken has been duly acknowledged and that complete thesis has been written by me. I understand the zero tolerance policy of the HEC and Government College University Faisalabad toward plagiarism. Therefore, I as an Author of above titled thesis declare that no portion of my thesis has been plagiarized and any material used as reference is properly referred/cited. I undertake that if I am found guilty of any formal plagiarism in the above titled thesis even after the award of PhD degree, the University reserves the rights to withdraw/revoke my PhD degree and that HEC and the University has the right to publish my name on the HEC/University website on which names of students are placed who submitted plagiarized thesis. Student/Author Signature Name: AsmaAziz iv v vi vii LIST OF CONTENTS Sr. No Title Page No. Declaration iii Plagiarism Undertaking iv Certificate by supervisory committee v Certificate by Scrutiny committee vi Certificate of Approval vii List of Contents viii List of Tables ix List of figures xiv List of abbreviations xviii Acknowledgements xix Abstract xx Chapter 1 Introduction 1 Chapter 2 Review of Literature 6 Chapter 3 Materials and Methods 26 Chapter 4 Results 39 Chapter 5 Discussion 118 Summary 132 Conclusions 137 Recommendations 138 References 139 viii LIST OF TABLES Table Title Page No. No. 1 Geographical and environmental conditions of the study sites. 27 2 Different populations of S. sarwari collected from the upstream and 30 downstream of four rivers (River Chenab, Ravi, Jhelum and Indus). 3 List of Chemicals used in the study. 32 4 List of RAPD primers used for the amplification of genome pattern in 36 RAPD markers. 5 Composition of PCR reagents for the amplification of genomic DNA 37 of S. sarwari. 6 PCR A/B used for the amplification of genome pattern in RAPD 37 markers. 7 Mean± S.E of different morphological characters of S. sarwai 45 collected from the different Rivers of the Punjab 8 Mean± S.E of different morphological characters of S. sarwai 47 collected from the Upstream of different Rivers of the Punjab. 9 Mean± S.E of different morphological characters of S. sarwai 48 collected from the downstream of different Rivers of the Punjab. 10 The Analysis of variance of different morphometric parameters of S. 51 sarwari collected from the different Rivers of the Punjab. 11 The Analysis of variance of different morphometric parameters of S. 53 sarwari collected from the upstream and downstream of different ix Rivers of the Punjab. 12 Mean± S.E of different meristic parameters of S.sarwai collected from 55 the different Rivers of the Punjab. 13 Mean ± S.E of different meristic parameters of S. sarwai collected 57 from the upstream of different Rivers of the Punjab. 14 Mean± S.E of different meristic characters of S.sarwai collected from 57 the downstream of different Rivers of the Punjab. 15 The Analysis of variance of different meristic characters of S. sarwari 58 collected from the different Rivers of the Punjab. 16 Comparison of the morphological parameters of S. sarwari with 60 respect to the upstream (US) and the downstream (DS) of the River Chenab. 17 Comparison of the morphological parameters of S. sarwari with 62 respect to the upstream (US) and the downstream (DS) of the River Jhelum. 18 Comparison of the morphological parameters of S. sarwari with 64 respect to the upstream (US) and the downstream (DS) of the River Ravi. 19 Comparison of the morphological parameters of S. sarwari with 66 respect to the upstream (US) and the downstream (DS) of the River Indus. 20 Quantification of isolated DNA with different methods from muscle 72 tissue of S. sarwari collected from different rivers of the Punjab, Pakistan. 21 Optimization and standardization of RAPD (SS) markers for the 76 x amplification of S. sarwari genome. 22 Description of amplicons in genomic DNA with polymorphism, gene 79 pool frequency and range size of 10 primers on different population of S. sarwari collected from different rivers of the Punjab, Pakistan. 23 Description of genetic indices; observed alleles (na), effected alleles 86 (ne), Nei’ diversity (h) and Shannon index (I) in the upstream population of S. sarwari of the River Chenab. 24 Description of genetic indices; observed alleles (na), effected alleles 87 (ne), Nei’s diversity (h) and Shannon index (I) of the downstream population of S. sarwari of the River Chenab. 25 Description of genetic indices; observed alleles (na), effected alleles 89 (ne), Nei’s diversity (h) and Shannon index (I) of the upstream population of S. sarwari of the River Jhelum. 26 Description of genetic indices; observed alleles (na), effected alleles 90 (ne), Nei’s diversity (h) and Shannon index (I) of the downstream population of S. sarwari of the River Jhelum. 27 Description of genetic indices; observed alleles (na), effected alleles 92 (ne), Nei’s (h) diversity and Shannon index (I) of the upstream population of S. sarwari of the River Ravi. 28 Description of genetic indices; observed alleles (na), effected alleles 93 (ne), Nei’s (h) diversity and Shannon index (I) of the downstream population of S. sarwari of the River Ravi. 29 Description of genetic indices; observed alleles (na), effected alleles 95 (ne), Nei’s diversity (h) and Shannon index (I) of the upstream population of S. sarwari of the River Indus. 30 Description of genetic indices, observed alleles (na), effected alleles 96 xi (ne), Nei’s diversity (h) and Shannon index (I) of the downstream population of S. sarwari of the River Indus. 31 Description of genetic indices; observed alleles (na), effected alleles 97 (ne), Nei’s diversity (h) and Shannon Index (I) of the upstream (US) and the downstream (DS) population of S. sarwari from four different rivers of the Punjab, Pakistan. 32 Estimation of Genetic variations (Hetrozygosity among population= 99 Ht; Diversity within population= Hs, Genetic variatons among populations= Gst and Genetic flow= Nm) among and between the eight populations of S. Sarwari collected from different upstream and downstream of the four rivers of the Punjab, Pakistan. 33 Genetic distance (d) and genetic similarity (S) among eight 101 populations of S. sarwari collected from the upstream (US) and the downstream (DS) of the River Chenab (C), Jhelum (J), Ravi (R) and Indus (I) based on the construction of Phylogenetic relationships by using POPGENE 32. 34 Description of genetic indices, observed alleles (na), effected alleles 105 (ne), Nei’s diversity (h) and Shannon index (I) of the population of S. sarwari of the River Chenab. 35 Description of genetic indices; observed (na), effected alleles (ne), 107 Nei’s diversity (h) and Shannon index (I) of the River Jhelum population of S. sarwari. 36 Description of genetic indices; observed alleles (na), effected alleles 108 (ne), Nei’s (h) diversity and Shannon index (I) of S. sarwari population of the River Ravi. 37 Description of genetic indices, observed (na), effected alleles (ne), 110 Nei’s and Shannon diversity (I) of the Indus population of S. sarwari. xii 38 Comparison of genetic indices, in population of S. sarwari collected 111 from the four different rivers of the Punjab. 39 Description of genetic variations (Hetrozygosity among population= 113 Ht; Diversity within population= Hs, Genetic variations among populations= Gst and Genetic flow= Nm) among and between the four populations of S. sarwari collected from four rivers (River Chenab, Jhelum, Ravi and Indus) of the Punjab, Pakistan. 40 Overall genetic distance (d) and genetic similarities (S) among 115 population of S. sarwari collected from four different rivers (River Chenab, Jhelum, Ravi and Indus) based on the construction of Phylogenetic relationships. xiii LIST OF FIGURES Fig. No. Title Page No 1 Map of Study Area 28 2 Morphology of S. sarwari (A&B) 29 3 The specimen of S. sarwari collected from the upstream 41 of Head Tarimue (River Chenab) showing light and dark color in adipose fin with black margin and black spot. 4 The specimen of S. sarwari collected from the 41 downstream of Head Tarimue (River Chenab) showing dark color in adipose fin with black margin and black spot. 5 The specimen of S. sarwari collected from the upstream 42 of Head Rasul (River Jhelum) showing brown and light gray color in adipose fin with dark gray margin and black spot.
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