Assessment and use of sea buckthorn and apple diversity in northern Pakistan Muhammad Arslan Nawaz Dissertation submitted for the academic degree Doktor der Agrarwissenschaften (Dr. agr.) Dissertation presented to the Faculty of Organic Agricultural Sciences Organic Plant Production and Agroecosystems Research in the Tropics and Subtropics (OPATS) University of Kassel 1st supervisor: Prof. Dr. Andreas Bürkert, University of Kassel 2nd supervisor: Prof. Dr. Asif Ali Khan, Muhammad Nawaz Shareef University of Agriculture, Multan Co-supervisor: Dr. Martin Wiehle, University of Kassel Date of submission: 03.12.2020 Date of defence: 15.04.2021 Witzenhausen, Germany Table of Content Acknowledgments .................................................................................................................. I Summary ............................................................................................................................. III Zusammenfassung .............................................................................................................. VI Chapter 1 - General introduction and research objectives ..................................................... 1 1.1 Perennial plant genetic resources – their importance and management ...................... 2 1.2 PGR in Pakistan and its mountainous regions ............................................................. 4 1.3 PGR in Gilgit and Baltistan .......................................................................................... 5 1.3.1 Topography, climate, and vegetation ..................................................................... 5 1.3.2 Neglected research region and threats to local PGR ............................................. 7 1.4 State of the art of assessing agrobiodiversity ............................................................... 9 1.5 Investigated local plant species ................................................................................. 11 1.5.1 Sea buckthorn (Hippophae rhamnoides L.) ......................................................... 11 1.5.2 Apple (Malus × domestica Borkh.) ....................................................................... 12 1.6 Research hypotheses and objectives ........................................................................ 14 1.7 Outline of the thesis ................................................................................................... 15 1.8 References ................................................................................................................ 17 Chapter 2 – Morphological and genetic diversity of sea buckthorn in the Karakoram Mountains of northern Pakistan ........................................................................................... 23 2.1 Abstract ..................................................................................................................... 24 2.2 Introduction ................................................................................................................ 25 2.3 Materials and Methods .............................................................................................. 28 2.3.1 Plant Material and Sampling Area ....................................................................... 28 2.3.2 Dendrometric Traits ............................................................................................. 30 2.3.3 Leaf and Fruit Morphometry Traits ...................................................................... 30 2.3.4 Leaf and Fruit Colours ......................................................................................... 30 2.3.5 DNA Isolation and EST-SSR Analysis ................................................................. 32 2.3.6 Data Analysis ...................................................................................................... 33 2.4 Results ...................................................................................................................... 35 2.4.1 Dendrometric Traits ............................................................................................. 35 2.4.2 Leaf and Fruit Morphometry Traits ...................................................................... 38 2.4.3 Leaf and Fruit Colour .......................................................................................... 38 2.4.4 Genetic Diversity and Differentiation of Markers .................................................. 39 2.4.5 Genetic Diversity and Differentiation of Populations ............................................ 42 2.5 Discussion ................................................................................................................. 46 2.5.1 Morphological Variation ....................................................................................... 46 2.5.2 Genetic Diversity and Differentiation ................................................................... 48 2.6 Conclusions ............................................................................................................... 50 2.7 Appendix ................................................................................................................... 52 2.8 References ................................................................................................................ 53 Chapter 3 – Superfruit in the niche — Underutilized sea buckthorn in Gilgit-Baltistan, Pakistan .............................................................................................................................. 60 3.1 Abstract ..................................................................................................................... 61 3.2 Introduction ................................................................................................................ 62 3.3 Materials and Methods .............................................................................................. 66 3.3.1 Study Area .......................................................................................................... 66 3.3.2 Survey on Sea Buckthorn Management, Harvest, and Post-Harvest Handling Practices ...................................................................................................................... 67 3.3.3 On-Farm Drying and Laboratory Experiment ....................................................... 69 3.3.4 Statistical Analysis .............................................................................................. 71 3.4 Results ...................................................................................................................... 72 3.4.1 Survey on Sea Buckthorn Management, Harvest, and Post-Harvest Handling Practices ...................................................................................................................... 72 3.4.1.1 Status and Subsistence through Collection ................................................... 72 3.4.1.2 Conditions and Methods of Harvest to Post-Harvest Handling ...................... 73 3.4.1.3 Collectors´ Problems .................................................................................... 74 3.4.1.4 Collectors’ Perception about Trainings.......................................................... 76 3.4.1.5 Preferred Plant and Fruit Traits by Collectors ............................................... 76 3.4.1.6 Sustainable Supply Chain Constructs ........................................................... 77 3.4.2 On-Farm Drying and Laboratory Experiment ....................................................... 78 3.4.2.1 Days Required for Drying and Moisture Content ........................................... 78 3.4.2.2 Concentration of Ascorbic Acid ..................................................................... 79 3.5 Discussion ................................................................................................................. 80 3.5.1 Status of Collection, Post-Harvest Handling, and Management ........................... 80 3.5.2 Quality Loss Due to Sun Drying of Sea Buckthorn Berries .................................. 82 3.5.3 Sea Buckthorn Supply Chain............................................................................... 83 3.6 Conclusions ............................................................................................................... 85 3.7 Appendix ................................................................................................................... 87 3.8 References ................................................................................................................ 89 Chapter 4 - Pheno-genetic studies of apple varieties in northern Pakistan: A hidden pool of diversity? ............................................................................................................................. 97 4.1 Abstract ..................................................................................................................... 98 4.2 Introduction ................................................................................................................ 99 4.3 Materials and Methods ............................................................................................ 102 4.3.1 Study site, plant material, and owner interview .................................................. 102 4.3.2 Dendrometry, management, and use ................................................................ 104 4.3.3 Fruit phenotypic description and phenotype .....................................................