Regeneration of a phosphate polluted field site [Elektronische Ressource] : implications of arbuscular mycorrhiza / von Verena Blanke

Regeneration of a
phosphate-polluted field site:
Implications of arbuscular mycorrhiza





Dissertation


zur Erlangung des akademischen Grades
doctor rerum naturalium (Dr. rer. nat.)








vorgelegt dem Rat der Biologisch-Pharmazeutischen Fakultät
der Friedrich-Schiller-Universität Jena



von Diplom-Biologin Verena Blanke
geboren am 11. August 1975 in Detmold




Jena, 02. April 2007







































Gutachter:
1.: Prof. Dr. François Buscot (Leipzig/ Halle)
2.: Prof. Dr. Stefan Halle (Jena)
3.: Prof. Dr. Edith Allen (Riverside, USA)
Tag der Doktorprüfung: 25. Juni 2007
Tag der öffentlichen Verteidigung: 13. Juli 2007







„Se. Excellenz der Herr Minister für Landwirthschaft, Domänen und
Forsten hat, um die Zucht der Trüffel im Königreiche Preussen nach
Möglichkeit zu fördern, mich beauftragt zunächst durch wissenschaftliche
Untersuchungen über die Bedingungen des Vorkommens und der
Entwickelung dieser Pilze der Angelegenheit planmässig näher zu treten.
[…] Es betrifft die Thatsache, dass gewisse Baumarten […] ganz
regelmässig sich im Boden nicht selbständig ernähren, sondern überall in
ihrem gesammten Wurzelsystem mit einem Pilzmycelium in Symbiose
stehen, welches ihnen Ammendienste leistet und die ganze Ernährung des
Baumes aus dem Boden übernimmt. So überraschend dieser Satz klingen
mag, so ist er durch die Ausdehnung meiner Untersuchungen schon jetzt
als festbegründet zu betrachten. […] Der ganze Körper ist also weder
Baumwurzel noch Pilz allein, sondern ähnlich wie der Thallus der Flechten,
eine Vereinigung zweier verschiedener Wesen zu einem einheitlichen
morphologischen Organ, welches vielleicht passend als Pilzwurzel,
Mycorhiza bezeichnet werden kann.“


A. B. Frank, 1885

Contents


List of tables .……………………………………………………………………………...…..………........ 1
List of figures …………………………………………………………………………………………......... 2
1 Introduction ……………………………………………………………………………………………... 4
1.1 Mycorrhizal symbioses …………………………………………………………………………... 4
1.2 Arbuscular mycorrhiza (AM) ………………………………………………………………..…… 5
1.2.1 Morphology ………………………………………………………………………………... 5
1.2.2 Fungal phylogeny and species identification …………………………………………... 8
1.2.3 Ecology and function ……………………………………………………………………... 8
1.2.4 AM in plant communities …………………………………………………………….…… 10
1.3 The Steudnitz field site …………………………………………………………………………... 11
1.4 Aims and structure of this study ………………………………………………………………… 11
2 The power of regeneration: Lessons from the Steudnitz field site …………… 14
2.1 Introduction: Regeneration and restoration potentials of systems ………………………….. 14
2.2 Characteristics and history of the study site …………………………………………………... 15
2.3 Projects and their outcomes …………………………………………………………………….. 16
2.3.1 Abiotic filters ………………………………………………. 16
2.3.2 Biotic filters ………………………………………………… 17
2.4 Conclusions: Current state concerning regeneration of the field site ………………………. 19
3 Arbuscular mycorrhiza in phosphate-polluted soil: Diversity of fungi ………………………. 20
3.1 Introduction: Which arbuscular mycorrhizal fungi (AMF) are present in Steudnitz? ……… 20
3.2 Materials and methods …………………………………………………………………………... 21
3.2.1 Root and spore sampling ………………………………………………………………… 21
3.2.2 DNA extraction and nested PCR ………………………………………………………... 21
3.2.3 Cloning, sequencing and sequence analyses …………………………. 22
3.3 Results …………………………………………………………………………………………….. 24
3.4 Discussion: Consequences for species identity and diversity ………………. 24
3.4.1 Identified AMF species …………………………………………………………………… 24
3.4.2 Relation of diversity to ecological context ……………… 25
3.5 Conclusions: Current state concerning AMF diversity ……………………………………….. 26
4 AM in phosphate-polluted soil:
Nitrogen supply affects root colonization of Artemisia vulgaris …………………… 27
4.1 Introduction: Why are AMF present in Steudnitz? ……………………………………………. 27
4.2 Materials and methods …………………………………………………………………………... 28
4.2.1 Experiment 1: Nitrogen gradient ………………………………………………………… 28
4.2.1.1 Soil parameters ……………………. 28
4.2.1.2 Plant material ………………………………………………………………………. 28
4.2.1.3 Plant element concentrations …….. 29
4.2.1.4 Root colonization by AMF ………………………………………………………… 29
4.2.2 Experiment 2: Nitrogen fertilization …………………………………………………...... 29
4.2.2.1 Experimental design ………………………………. 29
4.2.2.2 Plant material, element concentrations and root colonization by AMF ……… 30
4.2.2.3 Above-ground biomass …………………………………………… 30
4.2.3 Statistics …………………………………………………… 30
4.3 Results …………………………………………………………….. 31
4.3.1 Experiment 1: Nitrogen gradient ………… 31
4.3.1.1 Soil parameters ……………………………………………………………………. 31
4.3.1.2 Plant element concentrations …….. 31
4.3.1.3 Root colonization by AMF ………………………………………………………… 31
4.3.1.4 Correlations between root colonization and plant element concentrations …. 33
4.3.2 Experiment 2: Nitrogen fertilization …..…………………………………………………. 34
4.3.2.1 Plant element concentrations …………………………………………………….. 34
4.3.2.2 Root colonization by AMF ………………………… 34
4.3.2.3 Correlations between root colonization and plant element concentrations …. 37
4.3.2.4 Above-ground biomass …………………………………………………………… 37
4.4 Discussion: Implications of N levels and N:P imbalances …………………………………… 38
4.4.1 Experiment 1: Nitrogen gradient ………………………………………………………… 38
4.4.2 Experiment 2: Nitrogen fertilization ……………………………………………………... 41
4.5 Conclusions: Current state concerning the impact of nitrogen on AM ……………………... 41
5 AM in phosphate-polluted soil:
Interrelations between root colonization and nitrogen in four plant species ……. 42
5.1 Introduction: Which role do AMF play in Steudnitz? ….……………………………………… 42
5.2 Materials and methods …………………………………………………………………………... 43
5.2.1 Experiment 3: Fertilizer & fungicide, experimental design …………………………… 43
5.2.2 Plant material ……………………………………………………………………………… 43
5.2.3 Soil parameters, plant element concentrations, and root colonization by AMF ……. 43
5.2.4 Above-ground biomass …………………………………………………………………... 44
5.2.5 Statistics …………………………………………………………………………………… 45
5.3 Results …………………………………………………………….. 45
5.3.1 Soil Parameters …………… 45
5.3.2 Root colonization by AMF ……………………………………………………………….. 45
5.3.3 Plant element concentrations ……………………………. 47
5.3.4 Correlations between root colonization and soil parameters or plant element
concentrations …………………………………………………. 50
5.3.5 Above-ground biomass …………………………………………………………………... 52
5.4 Discussion: Consequences of reduced mycorrhization intensity …………………………… 53
5.4.1 Interrelations between mycorrhizal colonization, soil and plant N & P ……………… 53
5.4.2 Treatment effects on other elements …………………………………… 55
5.4.3 Nitrogen effects on species’ biomass ……………………………………... 56
5.4.4 Possible implications of mycorrhizal dependency for differences in
plant species’ responses ……………………………………………………… 56
5.4.5 Potential involvement of mycorrhizal networks …57
5.5 Conclusions: Current state concerning the impact of AM on plant elements ……………... 58
6 General discussion ..…………………………………………………………………………………... 59
7 General conclusions …….…………………………………………………………………………….. 65
8 References ………………………………………………. 66
Summary ………………………………79
Zusammenfassung ………………………………………………………………………………………. 81
Appendix ……………………………………. 83
Kooperationen – cooperations
Danksagung
Ehrenwörtliche Erklärung
Curriculum Vitae
Publications and Conference contributions

List of tables
List of tables


Table 1: Different features of arbuscular mycorrhizas (AM) and Ectomycorrhizas (EM) …….….. 5
Table 2: Soil data for the three field plots of experiment 1 …………………………………………. 31
Table 3: Plant element concentrations and percentage root colonization of Artemisia vulgaris
by arbuscular mycorrhizal fungi for the three field plots of experiment 1 ……………….. 32
Table 4: Results of Spearman rank correlations between mycorrhizal root colonization of
Artemisia vulgaris and plant N or N:P across plots of experiment 1 …………………….. 33
Table 5: Plant element concentrations of Artemisia vulgaris for nitrogen-fertilized plots (+N)
and control plots (-N) of experiment 2 ……………………………………………………… 35
Table 6: Percentage root colonization of Artemisia vulgaris by arbuscular mycorrhizal fungi
and above-ground biomass dry weight of A. vulgaris for nitrogen-f