On the inheritance and mechanism of baculovirus resistance of the codling moth, Cydia pomonella (L.) [Elektronische Ressource] / Sabine Asser-Kaiser geb. Asser

Kelkenbe

Sujets

• Biologie

Informations

• Publié par: johannes_gutenberg-universitat_mainz
• Publié le: 01 janvier 2009
• Nombre de lectures: 43
• Langue: English
• Poids de l'ouvrage: 1 Mo

Extrait

“On the inheritance and mechanism of baculovirus resistance of
the codling moth, Cydia pomonella (L.)”





Dissertation
zur Erlangung des Grades
“Doktor der Naturwissenschaften”



am Fachbereich Biologie
der Johannes Gutenberg-Universität
in Mainz





Sabine Asser-Kaiser
geb. Asser
geb. am 03.08.1980 in Karlsruhe



Mainz, 2009


Dekan:
1. Berichterstatter:
2.
Tag der mündlichen Prüfung: 29.04.2010 CONTENTS
CONTENTS
Contents .............................................................................................................................I
List of abbreviations .......................................................................................................VI
Summary.........................................................................................................................IX
1 General introduction.................................................................................................1
1.1 Cydia pomonella as a pest in apple prodution .................................................. 1
1.1.1 Life cycle of codling moth........................................................................ 1
1.1.2 Control of codling moth............................................................................ 2
1.2 Baculoviruses....................................................................................................5
1.2.1 Introduction...............................................................................................5
1.2.2 Structure and Classification...................................................................... 5
1.2.3 Genome and gene expression ................................................................... 7
1.2.4 Cydia pomonella Granulovirus (CpGV) as a member of the genus
Betabaculovirus ........................................................................................................ 8
1.2.5 Pathology..................................................................................................8
1.2.6 Symptoms of CpGV infection ................................................................ 10
1.3 Resistance.......................................................................................................11
1.3.1 Definition of resistance........................................................................... 11
1.3.2 Resistance to baculoviruses .................................................................... 12
1.3.3 Occurrence of CpGV resistant codling moth populations in Europe ..... 14
1.4 Aims of this thesis........................................................................................... 15
2 Resistance of codling moth against Cydia pomonella granulovirus (CpGV) is
inherited by a single, dominant gene located on the Z-chromosome ............................. 18
ICONTENTS
2.1 Introduction.....................................................................................................18
2.2 Material and methods......................................................................................19
2.2.1 Virus........................................................................................................19
2.2.2 Test insects..............................................................................................19
2.2.3 Single pair crosses .................................................................................. 20
2.2.4 Homogenisation of CpR ......................................................................... 20
2.2.5 Bioassays................................................................................................21
2.2.6 Determination of the sex of individuals.................................................. 22
2.3 Results and Discussion...................................................................................23
3 Sex linkage of CpGV resistance in a heterogeneous field strain of the codling moth
Cydia pomonella (L.)...................................................................................................... 32
3.1 Introduction.....................................................................................................32
3.2 Materials and methods....................................................................................34
3.2.1 Insects34
3.2.2 Virus........................................................................................................34
3.2.3 Bioassays................................................................................................34
3.2.4 Single pair crosses .................................................................................. 35
3.3 Results.............................................................................................................36
3.3.1 Single pair crosses within CpR............................................................... 36
3.3.2 Hybrid crosses between CpS and CpR ................................................... 38
3.3.3 Backcrosses of the F1 generation to CpR............................................... 39
3.4 Discussion.......................................................................................................41
4 Resistance overcoming character of the Iranian isolate CpGV-I12 and testing on
cross resistance to Cry1Ab ............................................................................................. 47
IICONTENTS
4.1 Introduction.....................................................................................................47
4.2 Material and methods......................................................................................49
4.2.1 Test Insects.............................................................................................49
4.2.2 Virus........................................................................................................49
4.2.3 Bioassays with CpGV............................................................................. 49
4.2.4 ith Cry1Ab protein .............................................................. 50
4.2.5 Statistical analysis...................................................................................51
4.3 Results.............................................................................................................51
4.3.1 Bioassays with CpGV-M and CpGV–I12 .............................................. 51
4.3.2 Instar-specific mortality of CpS and CpR............................................... 52
4.3.3 Bioassays with Bacillus thuringiensis Cry1Ab protein .......................... 54
4.4 Discussion.......................................................................................................54
5 Baculovirus resistance in codling moth (Cydia pomonella L.) caused by early
block of virus replication ................................................................................................ 57
5.1 Introduction.....................................................................................................57
5.2 Materials and methods....................................................................................60
5.2.1 Insects60
5.2.2 Virus........................................................................................................61
5.2.3 Bioassays with fluorescent brightener 28 ............................................... 61
5.2.4 Intra-haemocoelar infection....................................................................62
5.2.5 Tissue specific virus replication ............................................................. 62
5.2.6 DNA isolation and quantitative PCR...................................................... 63
5.2.7 Haemolymph transfusion........................................................................63
5.3 Results.............................................................................................................63
IIICONTENTS
5.3.1 Bioassays with fluorescent brightener 28 ............................................... 63
5.3.2 Intra-haemocoelar infection....................................................................64
5.3.3 Replication of CpGV in different tissues................................................ 65
5.3.4 Haemolymph transfusion........................................................................67
5.4 Discussion.......................................................................................................68
6 Investigations on protein-protein interactions between PE38 of CpGV-I12 and host
proteins of a CpGV resistant codling moth strain........................................................... 72
6.1 Introduction.....................................................................................................72
6.1.1 Background and aim of the study ........................................................... 72
6.1.2 Yeast two-hybrid system ........................................................................ 73
6.2 Material and methods......................................................................................76
6.2.1 Test insects..............................................................................................76
6.2.2 Virus...............................................................