Symbiotic Fungi: Principles and Practice (Soil Biology)

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24 Auxin Production by Symbiotic Fungi 391 l Drying gas (nitrogen) pressure: 18 psi (pounds per square inch, equivalent to 124 kPa) at 250 C, nebulizing gas (air): 50 psi (345 kPa) l Pressure of collision gas (argon): 1.4 mTorr l Needle voltage: 4400 V l Shield voltage: 600 V l Capillary voltage: 40 V l The mass spectrometer was operated with negative Electrospray ionization in multiple reaction monitoring mode. l Mass transition for IAA: m/z 173.9/ m/z 130.0 (collision energy (CE) 9.0 eV) l Mass transition for D 5-IAA: m/z 178.8/ m/z 134.0 (CE 11.5 eV) A calibration curve of the ratio of peak areas of unlabeled standards to peak area of deuterium labeled IAA was used for quantification. Remark: Because IAA occurs in common media components, extracts of media in which no fungus had been cultured should always be analyzed as negative controls. If any IAA is found in the pure medium, its concentration has to be subtracted from the IAA concentrations in culture filtrates of the fungus. Obviously, the use of media which do not contain detectable amounts of IAA is preferable. 24.5 Examples of IAA Production by Fungi 24.5.1 Piriformospora indica Piriformospora indica is an endophytic fungus which increases plant growth in many plants of different families (Varma et al. 1999). In contrast to arbuscular mycorrhizal fungi, P. indica grows well on artificial media, and it is also able to colonize cruciferous plants including A. thaliana. In many interactions increased shoot growth was accompanied by increased root biomass. The mechanism by which this beneficial effect is achieved has long been debated. In our bioassay, A. thaliana inoculated with P. indica showed increased root branching and reduced main root length (‘‘bushy roots’’), whereas the roots of uninoculated plants reached the bottom of the Petri dish after 2 weeks (Figs. 24.2a, b). Similarly enhanced root branching was observed after inoculation of the mycorrhizal plant Lotus japonicus with P. indica (Figs. 24.2c, d). The phenotype resembled very much typical auxin effects. Therefore we compared the effect of pure externally applied auxin in different concentrations (Figs. 24.2e, f, for details see Sirrenberg et al. 2007) with the effect of P. indica culture filtrates and its ethyl acetate extracts. The pure culture filtrate as well as the extracts inhibited main root length while increasing root branching. The IAA content in the culture filtrate was determined by HPLC-MS and GC-MS to be 1.36 +/ 0.36 mM (after 4 weeks of growth in M+ medium at 23 C on a rotary shaker (Sirrenberg et al. 2007).
392 A. Sirrenberg et al. 24.5.2 Truffles Truffles are symbiotic fungi forming ectomycorrhizae with trees (e.g., oaks, hazels) and some shrubs (e.g., Cistus). Production of IAA by ectomycorrhizal fungi is wellknown, and IAA, probably with other signal molecules, might drive the ectomycorrhiza genesis (Barker and Tagu 2000). Using the bioassay described in Fig. 24.3a, we demonstrated that the exudates of two truffle species (Tuber borchii and T. melanosporum) modify the root architecture of Arabidopsis thaliana (i.e., root shortening, increased branching and root hair length: see Figs. 24.3b1–b3). IAA was quantified by HPLC-MS directly from the MS agar zone of the bioassay (Fig. 24.3a). The mycelium of T. borchii (strain ATCC 96540) produced 1.3 0.3 *10 7 M IAA after 10 days of growth in the dark (about ten times more than was found in control Petri dishes containing no fungus). After establishing a dose response of A. thaliana root length to synthetic IAA, we concluded that the IAA exudated by T. borchii fully accounts for the root shortening of A. thaliana seedlings (Fig. 24.3c). References Barker SJ, Tagu D (2000) The roles of auxins and cytokinins in mycorrhizal symbioses. J Plant Growth Regul 19:144–154 Ek M, Ljungquist PO, Stenstrom E (1983) Indole-3-acetic acid production by mycorrhizal fungi determined by gas chromatography-mass spectrometry. New Phytol 94:401–407 Gruen HE (1959) Auxins and fungi. Annu Rev Plant Physiol 10:405–440 Delbarre A, Muller P, Imhoff V, Guern J (1996) Comparison of mechanisms controlling uptake and accumulation of 2,4-dichlorophenoxy acetic acid, naphtalene-1-acetic acid, and indol-3acetic acid in suspension-cultured tobacco cells. Planta 198:532–541 Sirrenberg A, Göbel C, Grond S, Czempinski N, Ratzinger A, Karlovsky P, Santos P, Feussner I, Pawlowski K (2007) Piriformospora indica affects plant growth by auxin production. Physiol Plant 131:581–589 Spaepen S, Vanderleyden J, Remans R (2007) Indole-3-acetic acid in microbial and microorganism-plant signaling. FEMS Microbiol Rev 31:425–448 Stasinopoulos TC, Hangarter RP (1990) Preventing photochemistry in culture media by long-pass light filters alters growth of cultured tissues. Plant Physiol 93:1365–1369 Taiz L, Zeiger E (2006) Plant physiology. Sinauer Associates, Sunderland, MA Yang Y, Zimmer UZ, Taylor CG, Schachtmann DP, Nielsen E (2006) High-affinity auxin transport by the AUX1 influx carrier protein. Curr Biol 16:1123–1127 Varma A, Verma S, Sudha, Sahay N, Bütehorn B, Franken P (1999) Piriformospora indica, a cultivable plant-growth-promoting root endophyte. Appl Environ Microbiol 65:2741–2744
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Soil Biology Volume 18 Series Edito
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Ajit Varma l Amit C. Kharkwal Edito
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Foreword So old, so new... More tha
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Foreword vii Little AE, Robinson CJ
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x Preface work in this challenging
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xii Contents 9 Role of Root Exudate
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Contributors L.K. Abbott School of
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Contributors xvii Falko Feldmann Ju
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Contributors xix K. Nara Asian Natu
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Contributors xxi Marc St-Arnaud Ins
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2 A. Das and A. Varma Fig. 1.1 Type
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4 A. Das and A. Varma the scientifi
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6 A. Das and A. Varma 1.3.2 Symbiot
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8 A. Das and A. Varma all the root
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10 A. Das and A. Varma 1. Such poly
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12 A. Das and A. Varma 1.3.3.9 Nitr
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14 A. Das and A. Varma about 1-2 mm
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16 A. Das and A. Varma fixed nitrog
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18 A. Das and A. Varma Mycorrhizae
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20 A. Das and A. Varma Fig. 1.6 Dia
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22 A. Das and A. Varma a b Root hai
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24 A. Das and A. Varma intracellula
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26 A. Das and A. Varma Becker A, Ni
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28 A. Das and A. Varma Trappe JM, B
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30 A. Jumpponen were once active in
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32 A. Jumpponen GA, USA) to avoid d
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34 A. Jumpponen 2.2.3.6 Cloning, Se
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36 A. Jumpponen Table 2.1 Fungi det
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38 A. Jumpponen of the community st
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40 A. Jumpponen Girvan MS, Bullimor
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42 I.A.F. Djuuna et al. 1991). Crop
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44 I.A.F. Djuuna et al. 3.2.2 Indir
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46 I.A.F. Djuuna et al. Mycorrhiza
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48 I.A.F. Djuuna et al. Boomsma CR,
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50 I.A.F. Djuuna et al. Saito M (19
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52 M. Giovannetti et al. evidenced
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54 M. Giovannetti et al. a c e Ster
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56 M. Giovannetti et al. 4.2.4 Rema
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58 M. Giovannetti et al. a b c Fig.
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60 M. Giovannetti et al. to 4.1 mm
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62 M. Giovannetti et al. The detect
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64 M. Giovannetti et al. Jones MD,
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66 A. Gobert and C. Plassard NO3 fl
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68 A. Gobert and C. Plassard After
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70 A. Gobert and C. Plassard with k
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72 A. Gobert and C. Plassard 10 9 2
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74 A. Gobert and C. Plassard Fig. 5
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76 A. Gobert and C. Plassard connec
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82 A. Gobert and C. Plassard a Rati
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84 A. Gobert and C. Plassard - upta
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86 A. Gobert and C. Plassard Table
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88 A. Gobert and C. Plassard Newman
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90 I.M. van Aarle as mycorrhizal hy
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92 I.M. van Aarle a wash buffer. Th
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94 I.M. van Aarle l Usually 20-50 m
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96 I.M. van Aarle Fig. 6.1 Extramat
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98 I.M. van Aarle A disadvantage of
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Chapter 7 In Vitro Compartmented Sy
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7 In Vitro Compartmented Systems to
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Chapter 8 Use of the Autofluorescen
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8 Use of the Autofluorescence Prope
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Chapter 9 Role of Root Exudates and
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9 Role of Root Exudates and Rhizosp
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Chapter 10 Assessing the Mycorrhiza
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10 Assessing the Mycorrhizal Divers
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178 D. Krüger et al. 8. Wash the p
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180 D. Krüger et al. Table 10.1 Ty
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182 D. Krüger et al. appear are in
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184 D. Krüger et al. tool such as
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186 D. Krüger et al. Ishikawa J, T
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188 D. Krüger et al. Sammeth M, Ro
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190 Z.M. Solaiman hyphae have been
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192 Z.M. Solaiman 11.2.2 Isolation
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194 Z.M. Solaiman 11.3 Conclusions
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Chapter 12 Interaction with Soil Mi
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12 Interaction with Soil Microorgan
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Chapter 13 Isolation, Cultivation a
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13 Isolation, Cultivation and In Pl
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228 K. Vogel-Mikusˇ et al. only be
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234 K. Vogel-Mikusˇ et al. STIM de
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236 K. Vogel-Mikusˇ et al. transfe
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Table 14.1 Element concentrations w
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240 K. Vogel-Mikusˇ et al. Fig. 14
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242 K. Vogel-Mikusˇ et al. Frey B,
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244 E. Dumas-Gaudot et al. TEMED N,
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246 E. Dumas-Gaudot et al. system i
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248 E. Dumas-Gaudot et al. Bromophe
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254 E. Dumas-Gaudot et al. by Bradf
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256 E. Dumas-Gaudot et al. 3. Take
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264 E. Dumas-Gaudot et al. Followin
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266 E. Dumas-Gaudot et al. Once hom
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268 E. Dumas-Gaudot et al. to both
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270 E. Dumas-Gaudot et al. were the
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272 E. Dumas-Gaudot et al. Dumas-Ga
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274 E. Dumas-Gaudot et al. Valot B,
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276 P.A. Olsson Fig. 16.1 Schematic
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278 P.A. Olsson Furthermore, C tran
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280 P.A. Olsson in AM fungi and a h
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282 P.A. Olsson (Graham et al. 1995
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284 P.A. Olsson Nakano A, Takahashi
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286 X.H. He et al. In many cases, N
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288 X.H. He et al. Table 17.1 Exper
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290 X.H. He et al. 17.3.1.1 Comment
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Chapter 18 Analyses of Ecophysiolog
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18 Analyses of Ecophysiological Tra
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308 I. Brito et al. fungi, little i
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310 I. Brito et al. 60% of moisture
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312 I. Brito et al. Thompson 1990;
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314 I. Brito et al. 19.8 Crop Rotat
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316 I. Brito et al. References Abbo
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318 I. Brito et al. Smith SE, Read
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320 F. Feldmann et al. inoculum of
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322 F. Feldmann et al. Table 20.1 B
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324 F. Feldmann et al. transferred
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326 F. Feldmann et al. 20.3.3 The A
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328 F. Feldmann et al. Inoculum is
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330 F. Feldmann et al. Fig. 20.5 Pr
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332 F. Feldmann et al. value for th
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334 F. Feldmann et al. of abiotic a
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336 F. Feldmann et al. Lackie SM, B
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338 M. Vestberg and A.C. Cassells b
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Page 437 and 438: Index A A. bisporus var. burnettii,
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Page 441 and 442: Index 427 Leghemoglobin, 11 Lentinu
Page 443 and 444: Index 429 Proteomics, 244 Protoplas
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