BRIEF NOTE Vesicular-Arbuscular Mycorrhizal Status of Spring Ephemerals in Two Ohio Forests1

BRENT G. DEMARS, Biology Department, Lakeland Community College, 7700 Clocktower Drive, Kirtland, OH 44094

ABSTRACT. Nineteen spring ephemerals (7 monocots and 12 dicots) were surveyed for vesicular-arbuscular mycorrhizae (VAM) in two Ohio forests during spring 1993 and 1994. Eleven of the nineteen species sampled displayed VAM development which was consistent between years and sites, except for con- catenata (). Patterns of VAM occurrence primarily reflected the taxonomic classification of the sampled species. In this study, all monocots sampled were mycorrhizal, while only 25% of the dicots developed VAM relationships. The occurrence of VAM in the dicots primarily reflected their as well. All Ranunculaceae species were mycorrhizal while those in the other five dicot families were not.

OHIO J. SCI. 96 (4/5): 97-99, 1996

INTRODUCTION In the field, sampling was conducted by extracting Approximately 95% of all species form mycorrhi- six individuals of each spring ephemeral species present zal associations (Trappe 1987). Of those mycorrhizal at each site per sampling event (see Table 1 for dates). associations, herbaceous species typically form the Following extraction, were placed in plastic bags vesicular-arbuscular mycorrhizal (VAM) type (Brundrett with soil intact for transport to the laboratory. Samples 1991)- The occurrence of VAM among herbaceous taxa were taken from the upland stand of each site as de- has been well documented in a variety of ecosystems scribed in Boerner and Koslowsky (1989) and DeMars including forests (Trappe 1987). However, surveys of and Boerner (1995) to avoid possible topographic varia- VAM occurrence within specific phenological groups tions. In the laboratory, roots were rinsed, separated, within forest ecosystems are scant (however, see cleared and stained using trypan blue (Phillips and Brundrett and Kendrick 1988). One phenological group Hayman 1970). Roots were preserved with a 1:1:10 FAA in forest ecosystems is the spring ephemerals. Spring (formalin:acetic acid:ethanol) solution and stored in ephemerals are species which complete their above plastic tissue cassettes under refrigeration. ground activity before canopy closure in moist, rich VAM development was examined in five randomly temperate forests throughout the world (Grubb and selected 2-3 cm root segments (including root tips) per Marks 1989). The purpose of this project was to examine sampled plant. VAM development was defined as the VAM occurrence in spring ephemerals in two Ohio presence of any internal nonseptate hyphae, vesicles, forests and to compare it with that reported in the lit- arbuscules, or chlamydospores. The percentage of root erature for taxa of other phenological groups in similar length colonized (PRLC) by mycorrhizal fungi was esti- forests. mated for each segment (Giovanetti and Mosse 1980). The mean PRLC was then computed for each plant and MATERIALS AND METHODS averaged for the entire sample (N = 6 plants per species Nineteen species of spring ephemerals from seven per sampling event). families were sampled in two Ohio forests during 1993 and 1994. One study site was the 40 ha Bohannan Pre- RESULTS AND DISCUSSION serve located in eastern Delaware County, OH (40°21'N, Eleven of the nineteen spring ephemeral species 82°56'W). It was an older growth, upland beech-maple examined for VAM development were mycorrhizal forest which has not been disturbed since approxi- (Table 1). The patterns of VAM development between mately 1900. Specific vegetation and soil information for sites and years were consistent and apparent in all the Bohannan Preserve is described in Boerner and cases. Consequently, statistical analysis was not deemed Koslowsky (1989). The other site was the Wright State Uni- necessary to describe obvious patterns among species. versity (WSU) woods located in northwestern Greene All the monocot species examined in this study were County (39°45'N, 84°03'W). This site was an older growth mycorrhizal (Table 1). The high percent root length (100+ years), oak-maple forest dominated by Acer colonized (PRLC) in the Liliaceae probably reflected saccharum Marsh., Quercus rubra L, and Quercus alba their root morphology. These species have magnolioid L. Specific vegetation and soil information is described root systems characterized by minimal root hair in DeMars and Runkle (1992) and DeMars and Boerner development (Baylis 1975). The low numbers of root (1995). hairs increase their dependence upon zygomycetous VAM resulting in mycotrophy (mycorrhizal depend- 'Manuscript received 4 April 1996 and in revised form 29 October ence) as described by Trappe (1987). Other studies 1996 (#BN96-08). have indicated similar results for some liliaceous spring 98 VAM STATUS OF SPRING EPHEMERALS VOL. 96

TABLE 1 ephemerals with magnolioid root systems; Brundrett and Kendrick (1988) observed 83% PRLC for Allium Mean percent root length colonized (PRLC) by vesicular-arbuscular tricoccum, 67% PRLC for Erythronium americanum, mycorrhizal fungi in spring ephemerals (N = 6 individuals per and 84% PRLC for Trillium erectum and Trillium gran- species per sampling event) in the Bohannan Woods Preserve (BW) diflorum in Ontario, while DeMars and Boerner (1995) and the Wright State University Woods (WSU). reported 60.2% PRLC for Trillium flexipes in Ohio. The distribution of VAM development in the dicots Spring Ephemeral Mean examined in this study reflected their taxonomic Species Family Site Date PRLC classification also. All Ranunculaceae species were mycorrhizal while only one sample from the other dicot Monocots families were (Table 1). Trappe (1987) indicated that the Allium tricoccum Ait. Liliaceae BW 4-22-93 56 4-25-94 64 order is highly mycotrophic as are most WSU 4-23-93 53 other terrestrial orders of the subclass Magnoliidea. The 4-19-94 57 exception to this is the Papaverales in which 6l% of all Erythronium albidum Nutt. WSU 4-23-93 63 species examined were scored as nonmycorrhizal 4-19-94 61 (Trappe 1987). In the present study both Erythronium americanum BW 4-22-93 59 canadensis and D. cucullaria (Fumariaceae/Papaverales) Ker Gawler 4-25-94 54 were nonmycorrhizal, lending support to the weakly Ornithogalum umbellatum L. # WSU 4-23-93 66 mycotrophic status of the order. Brundrett and Kendrick 4-19-94 52 (1988) also observed similar patterns for species they Trillium flexipes Raf. WSU 5-16-93 59 sampled in both these orders. 5-09-94 63 The orders (subclass Caryophyllidae) Trillium grandiflorum BW 5-23-93 50 and Capperales (subclass Dillieniidae) have also been (Michx.) Salisb. 5-19-94 61 deemed weakly mycotrophic by Trappe (1987). In the Trillium sessile L. BW 5-23-93 72 present study virginica (Portulaceae/ 5-19-94 59 Caryophyllales) and Cardamine spp. (Brassicaceae/ WSU 5-16-93 63 Capperales) were nonmycorrhizal with the exception of 5-09-94 67 Cardamine concatenata in the 1993 WSU sample. The Dicots nonmycorrhizal status of these species probably re- bulbosa (Michx.) Apiaceae BW 4-03-93 0 Nutt. 4-05-94 0 flected their more ruderal life histories and the pro- duction of mustard oil glucosides, glucosinolates, and Cardamine concatenata Brassicaceae BW 4-22-93 0 other defensive chemicals (Trappe 1987). However, it (Michx.) O. Schwarz 4-25-94 0 WSU 5-16-93 15 has not been definitively demonstrated that such phyto- 5-09-94 0 chemicals prevent the establishment of VAM (Schreiner Cardamine dissecta BW 4-22-93 0 and Koide 1993). (Leavenw.) Al-Shehbaz 4-25-94 0 The observation of VAM development in the 1993 Cardamine douglassii Britton BW 4-22-93 0 WSU sample of C. concatenata probably indicates an 4-25-94 0 "apparent" tendency to form VAM during its senescence WSU 4-23-93 0 period when root integrity becomes compromised Cardamine rhomboidea WSU 4-19-94 0 (DeMars and Boerner 1995). The formation of VAM in (Pers.) DC. the nonmycotrophic Brassicaceae has been reported by Fumariaceae BW 4-22-93 0 numerous authors (Harley and Harley 1987, Medve 1983, (Goldie) Walp. 4-25-94 0 Newman and Redell 1987, DeMars and Boerner 1994), Dicentra cucullaria BW 4-22-93 0 but its significance has often been questioned (Hirrel et (L.) Bernh. 4-25-94 0 al. 1978, Tester et al. 1987, Koide and Schreiner 1992) as Floerkia proserpinacoides Limnanthaceae BW 4-22-93 0 most authors suggest that it is nonfunctional due to its Willd 4-25-94 0 occurrence in senescent roots. Claytonia virginica L. Portulaceae BW 4-22-93 0 In the present study, Erigenia bulbosa (Apiaceae/ 4-25-94 0 Apiales) was nonmycorrhizal in all samples even though WSU 4-23-93 0 its family has shown a high degree of mycotrophy 5-16-93 0 4-19-94 0 (Trappe 1987). The nonmycorrhizal status in this species 5-09-94 0 may also reflect chemical defenses; however, cold Anemone virginiana L. Rannunculaceae BW 4-22-93 24 temperatures may have prevented VAM development. E. 4-25-94 34 bulbosa is the earliest spring ephemeral to become WSU 5-16-93 49 active. Its above ground activity is limited to March and 5-09-94 38 April in the study forests, a time when soil temperatures Anemonella thalictroides WSU 5-16-93 63 are low. Temperature has been shown to play a major (L.) Spach. 5-09-94 58 role in VAM development. Hayman (1983), Chilvers and Rannunculus ficaria L. # WSU 5-16-94 23 Daft (1982), and Anderson et al. (1987) have shown limited VAM development at temperatures as high as # indicates a non-native species. 10° C, and Smith and Bowen (1979) have reported lack OHIO JOURNAL OF SCIENCE B. G. DKMARS 99 of VAM initiation at slightly lower temperatures. phenology of plants in a sugar maple forest. Can. J. Bot. 66: In Floerkia proserpinacoides (Limnanthaceae/ 1153-1173. Chilvers, M. T. and M. J. F. Daft 1982 Effects of low temperature on Geraniales), the lack of VAM development may have development of the vesicular-arbuscular mycorrhizal association been a result of chemical defense compounds similar between Glomus caledonium and Allium cepa. Trans. Br. Mycol. to those found in members of the Brassicaceae. Gleason Soc. 79: 153-157. and Cronquist (199D reported that Floerkia sp. produces DeMars, B. G. andj. R. Runkle 1992 Groundlayer vegetation ordina- tion and site-factor analysis of the Wright State University woods mustard oils also. (Greene County, Ohio). Ohio J. Sci. 92: 98-106. Overall, the levels of PRLC in species of the myco- and R. E. J. Boerner 1994 Vesicular-arbuscular mycorrhizal fungi trophic taxa (Trappe 1987) corresponded well with colonization in Capsella bursa-pastoris (Brassicaceae). Am. Midi. Nat. 132: 377-380. those reported for other mycotrophic, forest spring and R. E. J. Boerner 1995 Mycorrhizal dynamics of three wood- ephemerals (Brundrett and Kendrick 1988, DeMars and land herbs of contrasting phenology along topographic gradients. Boerner 1995) and for summer-green herbs (Mayr and Am. J. Bot. 82: 1426-1431. Godoy 1989) such as Geranium robertianum L. (Boerner Giovanetti, M. and B. Mosse 1980 An evaluation of techniques for measuring vesicular-arbuscular mycorrhizal infection in roots. 1990), Geranium maculatum L. and Polygonatum New Phytol. 84: 489-500. pubescens (Walter) Elliot (Boerner 1986), and Smilacina Gleason, H. A. and A. Cronquist 1991 Manual of vascular plants of racemosa (L.) Desf. (Brundrett and Kendrick 1988, northeastern United States and adjacent Canada. New York Botanical DeMars and Boerner 1995). In conclusion, the my- Gardens, Bronx, NY. 910 pp. corrhizal status of spring ephemerals in the two sites in Grubb, P. J. and P. L. Marks 1989 Spring of eastern North American and European deciduous forests. Plants Today 2: 89-96. this study reflected their taxonomy. This is not surprising Harley, J. L. and E. L. Harley 1987 A check-list of mycorrhizas in the as mycotrophy (VAM dependence) originated from British flora. New Phytol. (Suppl.) 105: 1-102. revolutionary plant-fungal interactions in certain taxa Hayman, D. S. 1983 The physiology of vesicular-arbuscular endo- and from interactions which would have resulted in mycorrhizal symbiosis. Can. J. Bot. 6l: 944-963- Hirrel, M. C, H. Mehravaran, andj. W. Gerdemann 1978 Vesicular- preclusion of such fungi in other taxa (Trappe 1987). arbuscular mycorrhizae in the Chenopodiaceae and Cruciferae: Do they occur? Can. 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