Symbiotic Fungi: Principles and Practice (Soil Biology)

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17 15 N Enrichment Methods to Quantify Two-Way 287 inorganic chemicals or organic compounds (up to 99.90 atom 15 N, %) is applied to plant tissues, growth media or soils, and the events of N metabolism or N translocation are followed over short-term periods (days to weeks). The 15 N labeling method involves a large enrichment of 15 N over background, making measurement of isotopic effects easy because the difference between the isotopic compositions of the source and the plant or the soil is large. In the 15 N natural abundance or d 15 N method, plant tissues are analyzed to determine 15 N at naturally occurring levels (d 15 N values, %). In the d 15 N method, there is a small enrichment of 15 N over background, making measurement of isotopic effects difficult. The difference between isotopic compositions of the source and the plant or soil is small, but does reflect naturally occurring 15 N variations over long-term periods (years to decades) (Hobbie and Hobbie 2006). However, variations of d 15 N values are often small ( 10%) among AM or EM plant species that have unique physiological processes in different habitats (Handley and Scrimgeour 1997; Högberg 1997; Hobbie and Hobbie 2006). As a consequence, it is hard to demonstrate plantto-plant N transfer through CMNs by d 15 N variations of mychorrhizal plants, at least at face value, although they may demonstrate the magnitude of 15 N fractionations during N translocation in long-term natural conditions. 17.3 Experimental Design for Investigating Two-Way Nitrogen Transfer Between Plants Here we explore the 15 N enrichment method to determine two-way N transfer between plants through mychorrhizal connections and to measure the net benefit to the partners. With either the N2-fixing or the non-N2-fixing plant as the N-donor, four reciprocal pairings are created to study two-way plant-to-plant N transfer, and to distinguish N transfer between the soil and the mychorrhizal pathway (Table 17.1). First, as a control, the non-nodulated/non-mychorrhizal pair monitors N transfer in the absence of both hyphal connections and N2 fixation. Second, the nodulated/non-mychorrhizal pair tests effects of N2 fixation on N transfer in the absence of hyphal connections. Third, the non-nodulated/mychorrhizal pair determines effects of hyphal connections on N transfer in the absence of N 2-fixation. Fourth, the nodulated/mychorrhizal pair examines effects of both mychorrhizal and N2 fixation on N transfer. To minimize mass flow and diffusion through soil pathways, two perforated Perspex plates (2.5 mm thick) are inserted in the middle of a 5 l plastic box (300 12 150 cm) to divide the box into two chambers with a 5-mm air gap (Fig. 17.1). On one side of the two plates, sheets of 25–37 mm nylon mesh are inserted on the inner side of the plates to allow only hyphal connections. To further minimize water movement across the air gap between the two chambers, high water holding capacity crystals [0.5% (w/w), RainSaver, Hortex Australia Pty. Ltd., NSW, Australia] are incorporated in the potting mix. External 15 N may be added directly to the N-donor side.
288 X.H. He et al. Table 17.1 Experimental design for two-way N transfer between N2-fixing and non-N2-fixing plants grown in two-chambered pots. Enriched 15 N compounds are supplied to the growth medium or the leaves of the N-donor plant. Time period of 15 N labeling (days to months) depends on physiological activity of plants species and experimental purposes Plant pair name Plant status Treatment status N-donor N-receiver Myc Nod Non-nodulated/non-mycorrhizal pairs N2-fixing Non-N2-fixing Non-N2-fixing N2-fixing Nodulated/non-mycorrhizal pairs N2-fixing Non-N2-fixing + Non-N2-fixing N2-fixing + Non-nodulated/mycorrhizal pairs N2-fixing Non-N2-fixing + Non-N2-fixing N2-fixing + Nodulated/mycorrhizal pairs N2-fixing Non-N2-fixing + + Non-N2-fixing N2-fixing + + Myc Mycorrhizal status; Nod Nodulation status 17.3.1 Quantification of Nitrogen Transfer Between Plants Nitrogen transfer has been quantified in three ways: percentage of Ntransfer (% Ntransfer, (17.1–17.3)); amount of N transferred (mg plant 1 , (17.4); and percentage of NDFT (% NDFT, N in the N-receiver derived from transfer, (17.5) and (17.6)) (Ledgard et al. 1985; Giller et al. 1991; Johansen and Jensen 1996). %Ntransfer ¼ 15 Ncontentreceiver 100=ð 15 Ncontentreceiverþ 15 NcontentdonorÞ ð17:1Þ where 15 Ncontentplant ¼ atom% 15 N excessplant total Nplant=atom% 15 N excesslabeled N and atom% 15 N excessplant ¼ atom% 15 Nplant after labeling ð17:2Þ atom% 15 Nplant background ð17:3Þ Ntransfer ¼ %Ntransfer total Ndonor=ð100 % NtransferÞ ð17:4Þ % NDFT ¼ Ntransfer 100=total Nreceiver ð17:5Þ In addition, % NDFT values can also be calculated as follows (Tomm et al. 1994): %NDFT ¼ðatom % 15 N excessreceiver=atom % 15 N excessdonorÞ 100 ð17:6Þ Equations (17.5) and (17.6) are equal if the 0.3663% 15 N of atmospheric N2 is subtracted from the measured atom % 15 N in (17.5).
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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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78 A. Gobert and C. Plassard any ti
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80 A. Gobert and C. Plassard Net fl
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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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230 K. Vogel-Mikusˇ et al. Fig. 14
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234 K. Vogel-Mikusˇ et al. STIM de
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340 M. Vestberg and A.C. Cassells M
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342 M. Vestberg and A.C. Cassells a
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348 M. Vestberg and A.C. Cassells M
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350 M. Vestberg and A.C. Cassells 2
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352 M. Vestberg and A.C. Cassells 2
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354 M. Vestberg and A.C. Cassells C
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356 M. Vestberg and A.C. Cassells G
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358 M. Vestberg and A.C. Cassells P
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360 M. Vestberg and A.C. Cassells V
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362 A. Baldi et al. the capabilitie
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364 A. Baldi et al. 22.2.3 Initiati
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366 A. Baldi et al. 2. Place inocul
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368 A. Baldi et al. 22.5 Analysis 2
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370 A. Baldi et al. 22.5.4 Phenylal
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372 A. Baldi et al. Nicholas JB, Jo
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374 A. Baldi et al. Among these, fu
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378 A. Baldi et al. 23.4 Analysis F
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380 A. Baldi et al. Chattopadhyay S
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382 A. Sirrenberg et al. physical c
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384 A. Sirrenberg et al. Fig. 24.1
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390 A. Sirrenberg et al. in Fig. 24
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392 A. Sirrenberg et al. 24.5.2 Tru
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394 K. Haselwandter and G. Winkelma
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404 E. Mohammadi Goltapeh et al. Fi
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408 E. Mohammadi Goltapeh et al. 26
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410 E. Mohammadi Goltapeh et al. co
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412 E. Mohammadi Goltapeh et al. Tw
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414 E. Mohammadi Goltapeh et al. Co
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416 E. Mohammadi Goltapeh et al. qu
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418 E. Mohammadi Goltapeh et al. 26
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420 E. Mohammadi Goltapeh et al. Ch
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Index A A. bisporus var. burnettii,
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Index 425 Dark septate root endophy
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Index 427 Leghemoglobin, 11 Lentinu
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Index 429 Proteomics, 244 Protoplas
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Symbiotic Fungi: Principles and Practice (Soil Biology)

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