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1
1 Introduction
1.1 Prunus avium – Wild cherry
1.1.1 Introduction of Prunus avium for its use in forestry
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1 INTRODUCTION
2
1.1.2 Propagation of Prunus avium by in vitro culture
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Fig. 1 P. avium plants during the propagation process (A) Microshoots on rooting medium, genotype Neptun; (B) Two rooted and one unrooted microshoot, genotype Achilleus (C); P.
avium genotype Neptun plants four weeks and (D) eight weeks after transfer to soil; (E) 25 year old offspring of a selected plus tree (genotype Fama) of the silvaSELECT® clone collection.
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Fig. 2 Principle steps in propagation of Prunus avium microcuttings by in vitro culture.
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1 INTRODUCTION
4
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1.2 Microbial Endophytes
1.2.1 Definition and general description
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1.2 Microbial Endophytes
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1.2.2 Bacterial endophytes
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1 INTRODUCTION
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1.2.3 Impact of bacterial endophytes on plants
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1 INTRODUCTION
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1.2.4 Role of endophytes in in vitro culture
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Fig. 3 Visual observation of endophytic bacterial growth in different plant tissue culture materials. (A) Tissue culture of Solanum sp. with bacteria growing out of the basal part of the microshoot (B) Endophytes emerging from the submerged parts of Helleborus microshoots (C) bacteria of Phalaenopsis tissue culture growing on the surface of the medium (D) Leaves of Prunus avium genotype Neptun on bacterial culture medium with white colonies of isolate N-I-2 (Rhodopseudomonas palustris) (E) Leaves of Prunus avium genotype Demeter on bacterial culture medium with yellow colonies of isolate D-I-1 (Microbacterium testaceum).
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“It seems there is an equilibrium of endophytes and plants that under certain circumstances may be unbalanced to the detriment of one of the partners”
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1 INTRODUCTION
10
1.2.5 Detection of bacterial endophytes
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1.2 Microbial Endophytes
11
Fig. 4 Workflow for the detection of endophytes by culture-dependent and culture-independent method as used in this thesis.
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1 INTRODUCTION
12
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1.3 Cooperation project: “New possibilities of characterization, regulation and utilization of bacterial endophytes in in vitro propagation systems of wild cherry (Prunus avium L.)”
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1.4 Thesis objectives
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1.4 Thesis objectives
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1 INTRODUCTION
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2 Publications and Manuscripts
2.1 Improved in vitro rooting of Prunus avium microshoots by a dark treatment and an auxin pulse.
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2 PUBLICATIONS AND MANUSCRIPTS
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Title
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Authors
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Abstract
7KH LQIOXHQFH RI D IRXU GD\V GDUN WUHDWPHQW DQG D K ,%$ SXOVH RQ LQ YLWUR URRW IRUPDWLRQ RI Prunus avium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Keywords
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1. Introduction
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2. Materials and Methods
2.1 Plant material
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2.3 Rooting experiment
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2.5 Statistical analysis
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3. Results and Discussion
3.1 Importance of in vitro rooting on acclimatization success
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Fig. 1 Effect of in vitro rooting on the acclimatization success. (A) Plantlets four weeks after transfer to the greenhouse, genotype Neptun. (B) Acclimatization rate seven weeks after transfer to the greenhouse. Different letters indicate significant differences between the groups within one genotype by Tukey’s range test (P < 0.05); Bars = mean of four repetitions; Error bar = SD; n = 4 repetitions, number of total individuals are given in the bars.
3.2 Induction of roots by an IBA pulse
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Fig. 2 Comparison of the percentage of rooted plants produced using the standard protocol (control), a four days dark treatment (dark) or an 48 h IBA pulse (IBA). The 3 repetitions are depicted as red dots (A), green triangle (B) and blue squares (C), each dot represents one vessel (except for B, representing the whole experiment). Boxplot with additional information on the mean (dashed line). Asterisks indicate significant differences between the treatments and the corresponding control by Dunnett’s test (*, ** and *** indicate P ≤ 0.05, P ≤ 0.01 and P ≤ 0.001, respectively); n = 20-21.
Inductionof roots by dark treatment
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Fig. 2 Examples of rooted microshoots of Prunus avium genotypes Neptun, Apollo and Demeter after four weeks on rooting medium. (A) – (C) cultivated with standard protocol; (D) – (F) cultivated on standard medium with 4 days dark treatment; (G) – (I) cultivated with a 48 h IBA pulse before transfer on hormone free rooting medium. Bars represent 2 cm.
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References
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2.2 Endophytic bacteria in plant tissue culture: differences between easy- and difficult-to-propagate Prunus avium genotypes.
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© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com Tree Physiology 34, 524–533
doi:10.1093/treephys/tpu027
Endophytic bacteria in plant tissue culture: differences between HDV\DQGGLIÀFXOWWRSURSDJDWHPrunus aviumJHQRW\SHV
Mona Quambusch1,3, Anna Maria Pirttilä2, Mysore V. Tejesvi2, Traud Winkelmann1 and Melanie Bartsch1
1Woody Plant and Propagation Physiology Section, Institute for Horticultural Production Systems, Leibniz Universitaet Hannover, Herrenhaeuser Strasse 2, 30419 Hannover, Germany; 2Department of Biology, University of Oulu, Oulu, Finland; 3Corresponding author (quambusch@baum.uni-hannover.de)
Received October 22, 2013; accepted March 7, 2014; published online May 7, 2014; handling Editor Ron Sederoff
The endophytic bacterial communities of six Prunus avium L. genotypes differing in their growth patterns during in vitro SURSDJDWLRQZHUHLGHQWLÀHGE\FXOWXUHGHSHQGHQWDQGFXOWXUHLQGHSHQGHQWPHWKRGV)LYHPRUSKRORJLFDOO\GLVWLQFWLVRODWHV IURPWLVVXHFXOWXUHPDWHULDOZHUHLGHQWLÀHGE\6U'1$VHTXHQFHDQDO\VLV7RGHWHFWDQGDQDO\]HWKHXQFXOWLYDEOHIUDFWLRQ RIHQGRSK\WLFEDFWHULDDFORQHOLEUDU\ZDVHVWDEOLVKHGIURPWKHDPSOLÀHG6U'1$RIWRWDOSODQWH[WUDFW%DFWHULDOGLYHUVLW\
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KeywordsDPSOLÀHGULERVRPDOU'1$UHVWULFWLRQDQDO\VLVEDFWHULDOHQGRSK\WHVLQYLWURFXOWXUHSODQWJURZWKSURPRWLQJ EDFWHULD6U'1$VHTXHQFLQJ
Introduction
Wild cherry (Prunus avium L.) timber is a valuable hardwood, which is used for the production of veneers and solid wood furniture.
The timber is highly valued in Europe due to the reddish color and ÀUPQHVVRIWKHZRRGFRPSDUDEOHWRWURSLFDOWUHHVOLNHPDKRJDQ\
(Kobliha 2002). Economically most desired are fast-growing trees with straight stems (Janßen et al. 2010). To optimize growth char- acteristics, single plants showing a desired phenotype have been VHOHFWHGDQGDUHGLVWULEXWHGXQGHUWKHWUDGHPDUNVLOYD6(/(&7®. 9HJHWDWLYH SURSDJDWLRQ RI WKHVH FHUWLÀHG JHQRW\SHV YLD LQ YLWUR propagation enables high multiplication rates and stable clonal plant material (0HLHU'LQNHOHWDO). To ensure genetic diver- sity in forestry, a set of 31 genotypes was registered and is being propagated in vitro for tree nurseries (Janßen et al. 2010).
Under commercial conditions, severe losses are observed during in vitro rooting and acclimatization of the P. avium geno- types. Propagation success is strongly dependent on the geno- W\SHDQGVKRZVKLJKÁXFWXDWLRQRYHUWKH\HDUV7KHVHSUREOHPV could not be overcome by manipulation of culture media or RWKHU JURZWK FRQGLWLRQV & 6FKQHLGHU SHUVRQDO FRPPXQLFD- tion), and one possible explanation for the variation could be the presence of differing endophytic bacterial populations.
Endophytes are bacteria or fungi that, during part of their life, can live inside plant tissue without eliciting symptoms of disease (Petrini 1991). Endophytic bacteria are frequently observed in plant in vitro cultures, both in commercial labora- WRULHVDQGLQVFLHQWLÀFVWXGLHVLeifert et al. 1991), and often affect the in vitro propagation of trees (Ulrich et al. 2008a).
They were previously treated mainly as contaminants that
Research paper
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