Symbiotically Modified Organisms: Non-Toxic Fungal Endophytes in Grasses

Supplementary Material

Symbiotically modified organisms: non-toxic fungal endophytes in grasses

Pedro E. Gundel1,2, Luis I. Pérez2, Marjo Helander3 and Kari Saikkonen1

1 MTT Agrifood Research Finland, Plant Production Research, FI-31600 Jokioinen, Finland

2 IFEVA (CONICET – Agronomy Faculty, Buenos Aires University), Av. San Martin 4453, C1417DSE, Buenos Aires, Argentina

3Section of Ecology, Department of Biology, University of Turku, FI-20014 Turku, Finland

Corresponding author: Gundel, P.E. ()

Table of contents:

1) Survey of grass-endophyte literature

2) List of papers obtained from the searching process

3)Tables reporting Q’s and Rosenthal failsafe numbers

1) Survey of grass-endophyte literature

Meta-analysis allows statisticallycombining and comparing results from independent experiments that are dealing with the same or similar research questions [S1,S2].We used Web of Science ® tosearch the literature using the following keywords: “safe”, or “novel”, or “non-toxic”, or “non-ergot alkaloid-producing”, and “endophyte”. The database was completed with articles found following backward and forward the citation routes of Web of Science results.In total, 72 articles were found focusing on plant ecology and physiology, agronomy, animal production, biochemistry and natural products (see full list of papers below). Seventeen of them were discarded because of lack of experimental data. The 55 data papers were classified into papers including data on endophyte–mediated (i) livestock performance (consumption, animal growth, and animal respiration), (ii) plant performance (aboveground biomass,belowground biomass, and tillering), (iii) mycotoxin production (four group of alkaloids: Ergots, Lolitrems,Lolines and Peramine),and (iv) ecological interactions (aboveground herbivores, belowground herbivores, and cover other plants in the community). In addition, few studies comparing fungal biomass betweenman-made (SE+) and natural (E+) infected plants were used to examine theendophyte performance (v).

From the experimental papers, we selected those comparing naturally infected (E+) plants, plants infected with selected endophytes (SE+) by man, or manipulatively endophyte-free plants (ME-) from the same grass population or cultivar, under the same treatment or in common garden. To be included in the meta-analyses, a study had to provide results as either (i) means, measure of variance, and sample sizes in numerical or graphical form, (ii) correlation coefficients, (iii) two × two contingency tables, or (iv) F-, chi-square-, or t- tests statistics and degrees of freedom (df). Data presented in figures was digitalized using GetData Graph Digitizer 2.24. Only 34 of the papers presented sufficient data for the meta-analysis. However, out of these 34 papers, there were five articles [articles 54, 55, 56, 57, 62, inlist of papers obtained from the searching process] on genetically modified (GM) endophytes which had been developed to study the effect of blocking a particular point in the pathway of ergot synthesis. Even though they provide valuable information on vertebrate and invertebrate animal responses to such endophytes, they were withdrawn from the meta-analysis since first, they are not commercialized, and second, they are a case of paratransgenesis (see Glossary). The twenty nine papers that met the criteria are indicated in references with asterisks (*).

Primary publications commonly included several separate experiments (e.g. bioassays withdifferent species, cultivars and/or parallel experiments conducted in laboratory, greenhouse or commongarden), which we considered as independent observations. Thus, we obtained 330 independent experiments onlivestock performance (73), plant performance (163), mycotoxin production (45), and ecological interactions (94). About 20% of the studies used a same control for multiple comparisons or measures. Due to the lack of true independence between the studies, a “publication level average” was used[S3].

All analyses were carried out with the MetaWin version 2.0 statistical software[S1]. Statistical information provided as t-, F- or P- values was used to compute Hedges’ d as described by Borenstein et al.(2009)[S3].Mean effect sizesandbootstrapped confidence intervals were calculated using random effect models and with 4999 iterations, respectively [S2]. Effect was considered significant if the CI did not embrace zero. The Rosenthal fail-safe number was calculated to address the file drawer problem [S4]. This estimates the number of studies with non-significant results needed to be included into the meta-analysis tochange results from significant to non-significant.The Rosenthal fail-safe numbers for the overall estimate indicates sufficient robustness of the analyses.For example, if the Rosenthal failsafe number is higher than 5*N+10, where N is the sample size, we can assume that the statistical results is robust. However, in most of the comparisons between grass cultivars, fungal strains, etc. have to be treated as normative comparisons of individual studies because of the low number of cases.

Supplementary References

S1 Rosenberg, M.S. et al., eds (2000) MetaWin: Statistical Software for Meta–Analysis. Version 2.0., Sinauer Associates,Sunderland, MA, USA

S2 Gurevitch, J. and Hedges, L.V. (1993) Meta-analysis: combining the results of independent studies in experimental ecology. In The design and analysis of ecological experiments (Scheiner, S. and Gurevitch, J., eds), pp. 378–398. Chapman & Hall,New York.

S3 Borenstein, M. (2009) Effect sizes for continuous data. In The handbook of research synthesis and meta-analysis (3rdedn) (Cooper, H., Hedges L. and Valentine J., Eds), pp. 221–35. New York: Russell Sage Foundation

S4 Rosenberg, M.S. (2005) The file-drawer problem revisited: A general weighted method for calculating fail-safe numbers in meta-analysis. Evolution 59(2), 464–468

2) List of papers obtained from the searching process. The papers that were used in the meta-analysisare indicated with asterisk.

Adcock, R.A. et al.(1997) Sample variation and resource allocation for ergot alkaloid characterization in endophyte-Infected tall fescue. Crop Sci. 37, 31─35
Anowarul Islam, M. et al.(2011) Production and economics of grazing rye–annual ryegrass and tall fescue systems. Agron. J. 103, 558–564
*Assuero, S.G. et al.(2000) Morphological and physiological effects of water deficit and endophyte infection on contrasting tall fescue cultivars. N. Z. J. Agric. Res. 43, 49─61
*Ball, O.J.-P. et al.(2006) Importance of host plant species, Neotyphodium endophyte isolate, and alkaloids on feeding by Spodopterafrugiperda (Lepidoptera: Noctuidae) larvae. J. Econ. Entomol. 99(4), 1462–1473
Ball, O.J.-P. et al.(2011) Endophyte isolate and host grass effects on Chaetocnemapulicaria (Coleoptera: Chrysomelidae) Feeding. J. Econ. Entomol. 104(2), 665–672
Barker, D.J. et al.(2005) Contrasting toxic-endophyte contamination between endophyte-free and nontoxic-endophyte tall fescue pastures. Crop Sci. 45, 616─625
Beck, P.A. et al.(2008) Animal performance and economic comparison of novel and toxic endophyte tall fescues to cool-season annuals. J. Anim. Scien. 86, 2043─2055
*Belesky, D.P. et al.(2008) Does endophyte influence resource acquisition and allocation in defoliated tall fescue as a function of microsite conditions? J. Exp. Bot. 63, 368─377
*Bluett, S.J. et al.(2003) Perennial ryegrass endophyte effects on plasma prolactin concentration in dairy cows. N. Z. J. Agric. Res. 46(1), 9–14
Bluett, S.J. et al.(2004) Effects of natural reseeding and establishment method on contamination of a novel endophyte-infected perennial ryegrass dairy pasture with other ryegrass/endophyte associations. N. Z. J. Agric. Res. 47(3), 333–344
*Bluett, S.J. et al.(2005) Effects of a novel ryegrass endophyte on pasture production, dairy cow milk production and calf liveweight gain. Aust J Exp. Agric. 45, 11–19
*Bluett, S.J. et al.(2005) Effects of perennial ryegrass infected with either AR1 or wild endophyte on dairy production in the Waikato. N. Z. J. Agric. Res. 48(2), 197–212
Bouton, J.H. (2007) The economic benefits of forage improvement in the United States. Euphytica 154, 263─270
Bouton, J.H. et al.(2001) Selection for persistence in endophyte-free Kentucky 31 tall fescue. Crop Sci. 41, 1026─1028
Bouton, J.H. et al. 2002. Reinfection of tall fescue cultivars with non-ergot alkaloid–producing endophytes. Agron. J. 94, 567–574
*Bultman T.L. et al.(2003) Isolate-dependent impacts of fungal endophytes in a multitrophic interaction. Oikos 102(3), 491–496
*Bultman, T.L. et al.(2008) Influence of genetic variation in the fungal endophyte of a grass on an herbivore and its parasitoid.Entomol. Exp. Appl.130, 173–180.
*Burke, J.M. et al.(2006)Reproductive characteristics of endophyte-infected or novel tallfescue fedewes. Livestock Sci. 104, 103–111
Burns, J.C. and Fisher D.S. (2006) Consumption and digestion of ‘Jesup’ tall fescue hays with a novel fungal endophyte, without an endophyte, or with a wild-type endophyte. Crop Sci. 46, 216–223
Burns, J.C. et al. (2006) Grazing Influences on mass, nutritive value, and persistence of stockpiled Jesup tall fescue without and with novel and wild-type fungal endophytes. Crop Sci. 46, 1898–1912
*Coffey, K.P.et al (2007) Cow and calf performance while grazing tall fescue pastures with either the wild-type toxic endophyte or a non-toxic novel endophyte. AAES Res. Series 553, 67─69
Crush, J.R. et al.(2004) Effect of different Neotyphodium endophytes on root distribution of a perennial ryegrass (Lolium perenne L.) cultivar. N. Z. J. Agric. Res. 47(3), 345–349
*Drewnoski, M.E. et al.(2009) Performance of growing cattle grazing stockpiled Jesup tall fescue with varying. J. Anim. Sci. 87, 1034–1041
*Drewnoski, M.E. et al.(2007) Agronomic performance of stockpiled tall fescue varies with endophyte infection status. Forage Grazinglands. doi:10.1094/FG-2007-1217-03-RS.
*Drewnoski, M.E. et al.(2009) Growth and reproductive performance of beef heifers grazing endophyte-free, endophyte-infected and novel endophyte-infected tall fescue. Livestock Sci.125(2), 254–260
Easton, H.S. (2007) Grasses and Neotyphodium endophytes: co-adaptation and adaptive breeding. Euphytica 154, 295─306
*Eerens, J.P.J. et al.(1998) Influence of the endophyte (Neotyphodiumlolii) on morphology, physiology, and alkaloid synthesis of perennial ryegrass during high temperature and water stress. N. Z. J. Agric. Res. 41(2), 219–226
*Eerens, J.P.J. et al.(1998) Influence of the ryegrass endophyte (Neotyphodiumlolii) in a cool-moist environment III. Interaction with white clover. N. Z. J. Agric. Res. 41(2), 201–207
Felitti, A. et al. (2006) Transcriptome analysis of Neotyphodium and Epichloë grass endophytes. Fungal Genet. Biol. 43, 465-475
*Fisher, D.S. and Burns J.C. (2008) Testing for variation in animal preference for Jesup tall fescue hays with wild-type, novel, or no fungal endophyte. Crop Sci. 48, 2026─2032
Flores, R. et al. (2007) Effects of fescue type and sampling date on the ruminal disappearance kinetics of autumn-stockpiled tall fescue. J. Dairy Sci. 90, 2883─2896
Flores, R. et al. (2008) Effects of fescue type and sampling date on the nitrogen disappearance kinetics of autumn-stockpiled tall fescue. J. Dairy Sci.91, 1597─1606
Gunter, S.A. and Beck, P.A. (2004) Novel endophyte-infected tall fescue for growing beef cattle. J. Anim. Sci. 82, E75–E82
Hill, N.S. and Brown, E. (2000) Endophyte viability in seedling tall fescue treated with fungicides. Crop Sci. 40, 1490─1491
Hill, N.S. and Roach, P.K. (2009) Endophyte survival during seed storage: endophyte–host interactions and heritability. Crop Sci. 49, 1425–1430
Hill, N.S. et al. (2002) Strain-specific monoclonal antibodies to a nontoxic tall fescue endophyte. Crop Sci. 42, 1627─1630
Hopkins, A.A. and Alison, M.W. (2006) Stand persistence and animal performance for tall fescue endophyte combinations in the South Central USA. Agron. J. 98, 1221─1226
Hopkins, A.A. et al. (2010) Agronomic performance and lamb health among several tall fescue novel endophyte combinations in the South-Central USA. Crop Sci. 50, 1552─1561
Hopkins, A.A. et al.(2011) Registration of ‘Texoma’ MaxQ II tall fescue. J. Plant Regist. 5, 14─18
*Hunt, M.G. and Newman, J.A. (2005) Reduced herbivore resistance from a novel grass–endophyte association. J. Appl. Ecol. 42, 762–769
*Johnson, J.M. et al.(2012)Steer and pasture responses for a novel endophyte tall fescue developed for the uppertransition zone. J. Anim. Sci. 90, 2402─2409
Ju, H.-J. et al. (2006) Temperature influences on endophyte growth in tall fescue. Crop Sci. 46, 404─412
*Keathley, C.P. and Potter, D.A. (2012) Arthropod abundance in tall fescue, Loliumarundinaceum, pastures containing novel ‘safe’ endophytes.J. Appl. Entomol. 136, 576–587
Lambert, M.G. et al.(2004) Advances in pasture management for animal productivity and health. N. Z. Vet. J. 52(6), 311─319
Lowe, K.F. et al.(2008) The effect of endophyte on the performance of irrigated perennial ryegrasses in subtropical Australia. Aust. J. Agric. Res. 59,567─577
Malinowski, D.P. et al.(2004) Evidence for copper binding by extracellular root exudates of tall fescue but not perennial ryegrass infected with Neotyphodium spp. endophytes. Plant Soil 267, 1–12
*Malinowski, D.P. et al.(2005) Obligatory summer-dormant cool-season perennial grasses for semiarid environments of the Southern Great Plains. Agron. J. 97, 147–154
Malinowski, D.P. et al.(2011) Competitive ability of tall fescue against alfalfa as a function of summer dormancy, endophyte infection, and soil moisture availability. Crop Sci. 51, 1282–1290
*Matthews, A.K. et al. (2005) Intake, digestion, and N metabolism in steers fed endophyte-free, ergot alkaloid-producing endophyte-infected, or nonergot alkaloid-producing endophyte-infected fescue hay. J. Anim. Sci. 83, 1179─1185
*Mersch, S.M. and Cahoon, A.B. (2012) Biomass and tiller growth responses to competition between Ky31 and MaxQFestucaarundinacea cultivars and response of Ky31 to exogenously applied liquid preparation of Neotyphodium coenophialum underglasshouse conditions. Grass Forage Sci. 67, 299–304
Nelson, C.J. and Burns, J.C. (2006) Fifty years of grassland science leading to change. Crop Sci. 46, 2204─2217
*Nihsen, M.E. et al.(2004) Growth rate and physiology of steers grazing tall fescue inoculated with novel endophytes. J. Anim. Sci. 82, 878–883
Oram, R. and Lodge, G. (2003) Trends in temperate Australian grass breeding and selection. Aust. J. Agric. Res. 54, 211─241
Panaccione, D.G. et al.(2001) Elimination of ergovaline from a grass–Neotyphodium endophyte symbiosis by genetic modification of the endophyte. PNAS 98(22), 12820–12825
Panaccione, D.G. et al.(2003) Biochemical outcome of blocking the ergot alkaloid pathway of a grass endophyte. J. Agric. Food Chem. 51(22), 6429–6437
Panaccione, D.G. et al.(2006) Effects of ergot alkaloids on food preference and satiety in rabbits, as assessed with gene-knockout endophytes in perennial ryegrass (Lolium perenne). J. Agric. Food. Chem. 54, 4582–4587
Panaccione, D.G. et al.(2006) Ergot alkaloids are not essential for endophytic fungus-associated population suppression of the lesion nematode, Pratylenchusscribneri, on perennial ryegrass. Nematology 8(4), 583–590
Parish, J.A. et al.(2003) Use of nonergot alkaloid-producing endophytes for alleviating tall fescue toxicosis in sheep. J. Anim. Sci. 81, 1316–1322
Parish, J.A. et al.(2003) Use of nonergot alkaloid-producing endophytes for alleviating tall fescue toxicosis in stocker cattle. J. Anim. Sci. 81, 2856–2868
*Pennell, C.G.L. et al.(2005) Occurrence and impact of pasture mealybug (Balanococcuspoae) and root aphid (Aploneuralentisci) on ryegrass (Lolium spp.) with and without infection by Neotyphodium fungal endophytes. N. Z. J. Agric. Res. 48, 329–337
Pennell, C.G.L. et al.(2010) Development of a bird-deterrent fungal endophyte in turf tall fescue. N. Z. J. Agric. Res. 53(2), 145─150
Potter, D.A. et al.(2008) Contribution of ergot alkaloids to suppression of a grass-feeding caterpillar assessed with gene knockout endophytes in perennial ryegrass. Entomol. Exp. Appl. 126, 138–147
*Rasmussen S. et al. (2007) High nitrogen supply and carbohydrate content reduce fungal endophyte and alkaloid concentration in Lolium perenne. New Phytol. 173, 787–797
*Realini, C.E. et al.(2005) Effect of endophyte type on carcass traits, meat quality, and fatty acid composition of beef cattle grazing tall fescue. J. Anim. Sci. 83, 430─439
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*Rudgers, R.A. et al.(2010) Managing plant symbiosis: fungal endophyte genotype alters plant community composition. J. Appl. Ecol. 47, 468–477
van Zijll de Jong, E. et al.(2008) Detection and characterisation of novel fungal endophyte genotypic variation in cultivars of perennial ryegrass (Lolium perenne L.). Aust. J. Agric. Res. 59, 214─221
*Vibart, R.E. et al.(2008) Persistence and botanical composition of Jesuptall fescue with varying endophyte status after five years of stockpiling and intensive winter grazing. Forage and Grazinglands doi:10.1094/FG-2008-0421-01-RS.
*Watson, R.H. et al.(2004) Productivity of cow-calf pairs grazing tall fescue pastures infected with either the wild-type endophyte or a nonergot alkaloid-producing endophyte strain, AR542. J. Anim. Sci. 82, 3388–3393
Williams, W.M. et al. (2007) Future options and targets for pasture plant breeding in New Zealand. N. Z. J. Agric. Res. 50(2), 223─248
Woodfield, D.R. and Easton, H.S. (2004) Advances in pasture plant breeding for animal productivity and health. N. Z. Vet. J. 52(6), 300─310
Zhang, Y. et al. (2006) Recent Molecular and genomic studies on stress tolerance of forage and turf grasses. Crop Sci. 46, 497─511

3) Tables reporting Qb’s and Rosenthal failsafe numbers for host, cultivar and strain of all the comparisons and response variables

Table S1a.Qb’s values for the comparison between SE+vs E+
Comparison/Category / Response variable / Factor / df / Qb / Prob(Chi-Square)
SE+vs E+
Animal performance / Consumption / Symbiosis
Cultivar
Strain
Tester
Animal respiration / Symbiosis / 1 / 3.000 / 0.465
Cultivar / 1 / 0.445 / 0.505
Strain / 1 / 1.023 / 0.312
Tester
Animal growth / Symbiosis / 1 / 1.041 / 0.307
Cultivar / 3 / 2.463 / 0.482
Strain / 1 / 0.445 / 0.505
Tester / 1 / 1.023 / 0.312
Plant performance / Aboveground biomass / Symbiosis / 1 / 0.320 / 0.571
Cultivar / 3 / 3.542 / 0.315
Strain / 3 / 3.602 / 0.308
Belowground biomass / Symbiosis
Cultivar
Strain
Tillering / Symbiosis
Cultivar
Strain
Mycotoxin production / Alkaloids / Main / 2 / 1.604 / 0.448
Peramine / Symbiosis
Cultivar
Strain
Lolines / Symbiosis
Cultivar
Strain
Ergots / Symbiosis
Cultivar
Strain
Lolitrems / Symbiosis
Cultivar
Strain
Ecological interactions / Aboveground herbivores / Symbiosis
Cultivar / 1 / 21.854 / <0.001
Strain
Belowground herbivores / Symbiosis / 1 / <0.001 / 0.982
Cultivar / 1 / <0.001 / 0.982
Strain / 3 / 12.142 / 0.006
Cover of other plants species in the community / Symbiosis / 1 / 4.028 / 0.045
Cultivar / 2 / 9.844 / 0.007
Strain / 2 / 4.053 / 0.132
Endophyte performance / Fungus biomass / Endophyte frequency / Symbiosis / 1 / 12.657 / <0.001
Cultivar
Strain / 2 / 26.514 / <0.001

aSignificant comparisons are in bold

Table S1b.Qb’s values for the comparison between SE+vs ME–
Comparison/Category / Response variable / Factor / df / Qb / Prob(Chi-Square)
SE+vsME–
Animal performance / Consumption / Symbiosis
Cultivar
Strain
Tester
Animal respiration / Symbiosis
Cultivar / 1 / 0.111 / 0.738
Strain / 1 / 0.111 / 0.738
Tester
Animal growth / Symbiosis / 1 / 2.727 / 0.098
Cultivar / 2 / 4.148 / 0.126
Strain / 1 / 2.677 / 0.102
Tester
Plant performance / Aboveground biomass / Symbiosis / 1 / 0.176 / 0.717
Cultivar / 3 / 1.928 / 0.587
Strain / 3 / 1.347 / 0.769
Belowground biomass / Symbiosis
Cultivar
Strain
Tillering / Symbiosis
Cultivar
Strain
Ecological interactions / Aboveground herbivores / Symbiosis
Cultivar / 1 / 46.729 / <0.001
Strain / 1 / 0.171 / 0.679
Belowground herbivores / Symbiosis / 1 / 0.778 / 0.377
Cultivar / 1 / 0.778 / 0.377
Strain / 3 / 7.761 / 0.051
Cover of other plants species in the community / Symbiosis / 1 / 4.833 / 0.028
Cultivar / 2 / 15.164 / <0.001
Strain / 2 / 6.653 / 0.036

aSignificant comparisons are in bold

Table S1c.Qb’s values for the comparison between ME–vs E+
Comparison/Category / Response variable / Factor / df / Qb / Prob(Chi-Square)
ME–vs E+
Animal performance / Consumption / Symbiosis
Cultivar
Strain
Tester
Animal respiration / Symbiosis
Cultivar / 1 / 11.980 / <0.001
Strain / 1 / 11.980 / <0.001
Tester
Animal growth / Symbiosis / 1 / 1.465 / 0.226
Cultivar / 2 / 2.324 / 0.313
Strain / 1 / 0.857 / 0.354
Tester
Plant performance / Aboveground biomass / Symbiosis / 1 / 7.610 / 0.005
Cultivar / 3 / 7.026 / 0.071
Strain / 3 / 0.176 / 0.674
Belowground biomass / Symbiosis
Cultivar
Strain
Tillering / Symbiosis
Cultivar
Strain
Ecological interactions / Aboveground herbivores / Symbiosis
Cultivar / 1 / 61.385 / <0.001
Strain
Belowground herbivores / Symbiosis / 1 / 1.186 / 0.276
Cultivar / 1 / 1.186 / 0.276
Strain / 3 / 4.353 / 0.226
Cover of other plants species in the community / Symbiosis / 1 / 0.149 / 0.699
Cultivar / 2 / 0.521 / 0.771
Strain / 2 / 3.144 / 0.208

aSignificant comparisons are in bold

Table S2a. Rosenthal failsafe numbers for the comparison between SE+ and E+
Comparison/Category / Response variable / Factor / Rosenthal failsafe / N / 5*N+10
SE+ vs E+
Animal performance / Consumption / Main / 160.3 / 18 / 100
Symbiosis / 75.6 / 10 / 60
Cultivar / 75.6 / 10 / 60
Strain / 75.6 / 10 / 60
Tester / 75.6 / 10 / 60
Animal respiration / Main / 0 / 8 / 50
Symbiosis / 0 / 8 / 50
Cultivar / 0 / 7 / 45
Strain / 0.5 / 8 / 50
Tester / 0 / 8 / 50
Animal growth / Main / 189.9 / 39 / 205
Symbiosis / 300.5 / 38 / 200
Cultivar / 304.3 / 37 / 195
Strain / 295.1 / 37 / 195
Tester / 298.8 / 38 / 200
Plant performance / Aboveground biomass / Main / 0 / 32 / 170
Symbiosis / 0 / 32 / 170
Cultivar / 0 / 30 / 160
Strain / 0 / 32 / 170
Belowground biomass / Main / 0 / 5 / 35
Symbiosis / 0 / 5 / 35
Cultivar / 0 / 5 / 35
Strain / 0 / 5 / 35
Tillering / Main / 0 / 3 / 25
Symbiosis / 0 / 3 / 25
Cultivar / 0 / 3 / 25
Strain / 0 / 3 / 25
Mycotoxin production / Alkaloids / Main / 49.9 / 14 / 80
Peramine / Symbiosis / 7.4 / 5 / 35
Cultivar / 7.4 / 5 / 35
Strain / 7.4 / 5 / 35
Lolines / Symbiosis / 1.9 / 6 / 40
Cultivar / 1.9 / 6 / 40
Strain / 1.9 / 6 / 40
Ergots / Symbiosis / 0 / 3 / 25
Cultivar / 0 / 3 / 25
Strain / 0 / 3 / 25
Lolitrems / Symbiosis
Cultivar
Strain
Ecological interactions / Aboveground herbivores / Main / 0 / 18 / 100
Symbiosis / 0 / 18 / 100
Cultivar / 0 / 17 / 95
Strain / 0 / 18 / 100
Belowground herbivores / Main / 0 / 11 / 65
Symbiosis / 0 / 11 / 65
Cultivar / 0 / 11 / 65
Strain / 0 / 11 / 65
Cover of other plants species in the community / Main / 52.2 / 28 / 150
Symbiosis / 59.7 / 28 / 150
Cultivar / 58.1 / 27 / 145
Strain / 58 / 28 / 150
Endophyte performance / Fungus biomass / Endophyte frequency / Main / 0 / 7 / 45
Symbiosis / 0 / 7 / 45
Cultivar / 0 / 6 / 40
Strain / 0 / 7 / 45
aUnbiased are indicated in bold
Table S2b. Rosenthal failsafe numbers for the comparison between SE+ and ME–
Comparison/Category / Response variable / Factor / Rosenthal failsafe / N / 5*N+10
SE+ vs ME–
Animal performance / Consumption / Main / 38.2 / 17 / 95
Symbiosis / 1.5 / 9 / 55
Cultivar / 1.5 / 9 / 55
Strain / 1.5 / 9 / 55
Tester / 1.5 / 9 / 55
Animal respiration / Main / 12.5 / 4 / 30
Symbiosis / 12.5 / 4 / 30
Cultivar / 9.8 / 4 / 30
Strain / 9.8 / 4 / 30
Tester / 12.5 / 4 / 30
Animal growth / Main / 0 / 12 / 70
Symbiosis / 0 / 11 / 65
Cultivar / 3.1 / 10 / 60
Strain / 2.9 / 9 / 55
Tester / 0 / 11 / 65
Plant performance / Aboveground biomass / Main / 194.2 / 31 / 165
Symbiosis / 187.0 / 31 / 165
Cultivar / 182.8 / 31 / 165
Strain / 200.6 / 30 / 160
Belowground biomass / Main / 11.7 / 5 / 35
Symbiosis / 11.7 / 5 / 35
Cultivar / 11.7 / 5 / 35
Strain / 11.7 / 5 / 35
Tillering / Main / 5.9 / 5 / 35
Symbiosis / 5.9 / 5 / 35
Cultivar / 5.9 / 5 / 35
Strain / 5.9 / 5 / 35
Ecological interactions / Aboveground herbivores / Main / 17.9 / 32 / 170
Symbiosis / 17.9 / 32 / 170
Cultivar / 65.9 / 31 / 165
Strain / 20.5 / 30 / 160
Belowground herbivores / Main / 0.4 / 11 / 65
Symbiosis / 0 / 11 / 65
Cultivar / 0 / 11 / 65
Strain / 0 / 11 / 65
Cover of other plants species in the community / Main / 106.1 / 28 / 150
Symbiosis / 109 / 28 / 150
Cultivar / 127.6 / 27 / 145
Strain / 112.1 / 28 / 150
aUnbiased are indicated in bold
Table S2c. Rosenthal failsafe numbers for the comparison between ME–vs E+
Comparison/Category / Response variable / Factor / Rosenthal failsafe / N / 5*N+10
ME– vs E+
Animal performance / Consumption / Main / 219.1 / 17 / 95
Symbiosis / 68.1 / 9 / 55
Cultivar / 68.1 / 9 / 55
Strain / 68.1 / 9 / 55
Tester / 68.1 / 9 / 55
Animal respiration / Main / 0.5 / 4 / 30
Symbiosis / 0.5 / 4 / 30
Cultivar / 16.9 / 4 / 30
Strain / 16.9 / 4 / 30
Tester / 0.5 / 4 / 30
Animal growth / Main / 0 / 11 / 65
Symbiosis / 0 / 10 / 60
Cultivar / 0 / 9 / 55
Strain / 0 / 9 / 55
Tester / 0 / 10 / 60
Plant performance / Aboveground biomass / Main / 113.4 / 27 / 145
Symbiosis / 133.8 / 27 / 145
Cultivar / 123.4 / 27 / 145
Strain / 128.6 / 27 / 145
Belowground biomass / Main / 9.9 / 7 / 45
Symbiosis / 9.9 / 7 / 45
Cultivar / 9.9 / 7 / 45
Strain / 9.9 / 7 / 45
Tillering / Main / 0 / 2 / 20
Symbiosis / 0 / 2 / 20
Cultivar / 0 / 2 / 20
Strain / 0 / 2 / 20
Ecological interactions / Aboveground herbivores / Main / 15.7 / 19 / 105
Symbiosis / 15.7 / 18 / 100
Cultivar / 33.5 / 18 / 100
Strain / 15.7 / 18 / 100
Belowground herbivores / Main / 3.1 / 11 / 65
Symbiosis / 2.2 / 11 / 65
Cultivar / 2.2 / 11 / 65
Strain / 1.7 / 11 / 65
Cover of other plants species in the community / Main / 0 / 28 / 150
Symbiosis / 0 / 28 / 150
Cultivar / 0 / 27 / 145
Strain / 0 / 28 / 150
aUnbiased are indicated in bold