Genetic marker analysis in potato for extreme resistance (Ry_1tns_1tnt_1tno) to PVY and for chip quality after long term storage at 4 C̊ [Elektronische Ressource] / Ye-Su Song

Msnc

Informations

• Publié par: technische_universitat_munchen
• Publié le: 01 janvier 2004
• Nombre de lectures: 23
• Langue: Deutsch
• Poids de l'ouvrage: 3 Mo

Extrait

Lehrstuhl für Pflanzenbau und Pflanzenzüchtung
der Technischen Universität München



Genetic marker analysis in potato
for extreme resistance (Ry ) to PVY sto
and for chip quality after long term storage at 4°C


Ye-Su Song


Vollständiger Abdruck der von der Fakultät Wissenschaftszentrum
Weihenstephan für Ernährung, Landnutzung und Umwelt
der Technischen Universität München zur Erlangung des akademischen
Grades eines

Doktors der Agrarwissenschaften

genehmigten Dissertation.



Vorsitzender: Univ.-Prof. Dr. W. H. Schnitzler
Prüfer der Dissertation: 1. Univ.-Prof. Dr. G. Wenzel
2. Univ.-Prof. Dr. G. Forkmann



Die Dissertation wurde am 26. 07. 2004 bei der Technischen Universität
München eingereicht und durch die Fakultät Wissenschaftszentrum
Weihenstephan für Ernährung, Landnutzung und Umwelt am 18. 10. 2004
angenommen.
































Contents I

Contents

Contents.............................................................................................................................................................. I
Abbreviations................................................................................................................................................. V

1 Introduction....................................................................................................................................... 1
1.1 Taxonomy and importance of the potato crop........................................... 1
1.2 Potato genetics....................................................................................................................... 2
1.3 Molecular marker development and mapping in potatoes.............. 5
1.4 Extreme resistance to PVY (Ry ) ........................................................................ 9 sto
1.5 Chip quality after long term storage at 4°C................................................... 12
1.6 Aims of this dissertation................................................................................................ 18

2 Materials and methods....................................................................................................... 20
2.1 Plant material........................................................................................................................... 20
2.1.1 Extreme resistance to PVY (Ry ).............................................................. 20 sto
2.1.1.1 Basic material.......................................................................................... 20
2.1.1.2 Population for marker development........................................ 20
2.1.1.3 Cultivars for marker selection...................................................... 20
2.1.1.4 Cultivars for marker proofing....................................................... 21
2.1.2 Chip color quality after long term cold storage.................................. 21
2.1.2.1 Basic material.......................................................................................... 21
2.1.2.2 Dihaploid population........................................................................... 21
2.1.2.3 Tetraploid population......................................................................... 22

2.2 General molecular marker analysis methods............................................. 22
2.2.1 Genomic DNA isolation....................................................................................... 22
TM
2.2.2 Amplified Fragment Length Polymorphism (AFLP )

analysis............................................................................................................................ 23
2.2.2.1 General information............................................................................ 23
2.2.2.2 Restriction-ligation reactions........................................................ 23
2.2.2.3 Pre-amplification and selective amplification................... 24
2.2.2.4 Gel electrophoresis and imaging.............................................. 26
Contents II

2.2.3 Simple Sequenced Repeat (SSR) analysis.......................................... 27
2.2.4 Restriction Fragment Length Polymorphism (RFLP)

analysis............................................................................................................................ 29
2.2.4.1 Probes.......................................................................................................... 29
2.2.4.2 Restriction enzyme reaction......................................................... 29
2.2.4.3 Gel analysis and blotting................................................................. 29
2.2.4.4 Probe DNA isolation........................................................................... 30
2.2.4.5 Probe DNA labeling............................................................................ 31
2.2.4.6 Hybridization............................................................................................ 32

2.3 Potato virus Y resistance test................................................................................... 32
2.3.1 Preliminary examinations................................................................................... 32
2.3.2 Infection tests.............................................................................................................. 33
2.3.2.1 Primary infection.................................................................................... 33
2.3.2.2 Analysis of secondary infections............................................... 33
2.3.3 Selection of resistant and susceptible lines......................................... 33

2.4 Ry marker development and mapping......................................................... 33 sto
2.4.1 Ry marker application..................................................................................... 33 adg
2.4.2 Bulked Segregant Analysis............................................................................... 34
2.4.3 Mapping of Ry ....................................................................................................... 34 sto
2.4.3.1 Polymorphism analysis..................................................................... 34
2.4.3.2 Linkage grouping.................................................................................. 35
2.4.4 Cloning of Ry markers..................................................................................... 35 sto
2.4.5 Development of a PCR marker based selection system............ 36
2.4.6 Pedigree analysis.................................................................................................... 36

2.5 Chip color quality after long term storage at 4°C.................................... 37
2.5.1 Evaluation of chip quality and related trials.......................................... 37
2.5.1.1 Field trials and storage..................................................................... 37
2.5.1.2 Determination of chip quality........................................................ 37
2.5.1.3 Measurement of reducing sugar content............................ 38
2.5.1.4 Determination of starch content................................................. 39

2.6 Construction of a genetic map for good chip color quality
after long term storage at 4°C................................................................................… 40
Contents III

2.6.1 Molecular marker analysis................................................................................ 40
2.6.1.1 Genetic analysis.................................................................................... 40
2.6.1.2 Segregation analysis.......................................................................... 41
2.6.1.3 Linkage group analysis..................................................................... 41

2.7 QTL-Analysis for good chip color quality after long term
storage at 4°C.......................................................................……............................................ 42
2.7.1 Data analysis............................................................................................................... 42
2.7.1.1 Normality test........................................................................................... 42
2.7.1.2 Correlation analysis............................................................................ 42
2.7.1.3 Simple marker regression analysis......................................... 42
2.7.1.4 Analys