166 AKAMA K. et al. 5g 0.5g 12gKH 2 PO 4 1g, MgSO 4 7H 2 O0.5 g 1000ml5 Table 1. Derivation of the fungi examined Elaphomyces 1 E. granulatus
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1 (Bulletin of FFPRI) Vol.7 No.4 (No.409) December 2008 研究資料 (Research material) 様々な培地上における外生菌根菌の成長様式 1)* 1) 1) Growth of ectomycorrhizal fungi on various culture media Keiko AKAMA 1)* Hiroaki OKABE 1) and Takashi YAMANAKA 1) Abstract Effects of temperature and culture medium on the growth of ectomycorrhizal (EM) fungi were studied. Initially, 62 isolates of EM fungi were cultured at various temperatures from 5 to 40. Most isolates grew well at 25 C, whereas no apparent growth was observed at 5, 10 or 40 C. Next, growth of 202 isolates of EM fungi was examined at 25 C on eight different media (Hagem medium, peptone-yeast medium, potato-dextrose medium, Kawai-Ogawa medium, modified Melin-Norkrans medium, three modifications of malt extract medium) by measureing colony diameter on agar media and dry weight of mycelia in liquid culture. Growth patterns of the fungi on these media were described at generic level. Key words : ectomycorrhizal fungi, growth, temperature, culture media Brundrett ,000 Molina 1992 Taniguchi 2006 Miller 1983Taylor1971 1) 外生菌根菌 Table Received 12 June Accepted 10 September ) Department of Forest Microbiology, Forestry and Forest Products Research Institute (FFPRI) * Department of Forest Microbiology, Forestry and Forest Products Research Institute (FFPRI), 1 Matsunosato, Tsukuba, Ibaraki , Japan; hirabuki@ffpri.affrc.go.jp
2 166 AKAMA K. et al. 5g 0.5g 12gKH 2 PO 4 1g, MgSO 4 7H 2 O0.5 g 1000ml5 Table 1. Derivation of the fungi examined Elaphomyces 1 E. granulatus E. sp E. mutabilis E. sp Cenococcum 5 C. geophilum Tuber 6 T. indicum T. borchii Hygrophorus 8 H. russula H. arbustivus Lyophyllum 10 L. fumosum L. shimeji L. sykosporum L. semitale Laccaria 14 L. amethystea L. amethystea L. nigra L. bicolor L. vinaceoavellanea L. laccata Tricholoma 20 T. saponaceum T. matsutake T. ustale T. flavovirens T. sejunctum T. orirubens T. vaccinum Armillaria 27 A. albonaripes A. ponderosa Amanita 29 A. muscaria A. pantherina A. sychnopyramis A. hemibapha subsp.hemibapha A. hemibapha subsp.javanica A. longistriata A. vaginata var.vaginata A. volvata A. citrina var.citrina A. rubescens A. rubrovolvata A. virosa A. pseudoporphyria A. spissacea A. excelsa A. esuculenta A. spreta A. neovoidea
3 Growth of ectomycorrhizal fungi on various culture media 167 Table 1. Derivation of the fungi examinedcontinue 47 A. sinensis A. verna A. virgineoides A. abrupta A. hongoi A. flavipes A. gymnopus A. citrina var. grisea Cortinarius 55 C. purpurascens C. pseudopurpurascens C. bolaris C. pseudosalor C. subarmilatus Dermocybe 60 D. cinnamomea Hebeloma 61 H. crustuliniforme H. sacchariolens H. sacchariolens H. crustuliniforme f.microspermum Rhodophyllus 65 R. clypeatus Paxillus 66 P. involutus P. sp Choogomphus 68 C. tomentosus Boletinus 69 B. paluster B. cavipes Suillus 71 S. pictus S. laricinus S. bovinus S. granulatus S. granulatus S. grevillei S. luteus S. placidus S. variegatus S. spectabilis Phylloporus 81 P. bellus var.cyanescens Gyroporus 82 G. sp Gyrodon 83 G. lividus Xerocomus 84 X. subtomentosus X. chrysenteron X. nigromaculatus X. obscurebrunneus Chaliciporus 88 C. piperatus Bulletin of FFPRI Vol.7 No
4 168 AKAMA K. et al. Table 1. Derivation of the fungi examinedcontinue Pulveroboletus 89 P. ravenelii Aureoboletus 90 A. thibetanus Xanthoconium 91 X. affine Boletus 92 B. fraternus B. pulverulentus B. reticulatus B. ornatipes B. vilaceofuscus B. calopus B. luridus B. pseudocalopus B. erythropus B. granulopunctatus B. griseus Tylopilus 103 T. chromapes T. neofelleus T. nigropurpureus T. valens T. ferrugineus T. eximuis T. virens T. nigerrimus Leccinum 111 L. scabrum L. extremiorientale L. versipelle L. hortonii L. holopus L. intusrubens L. insigne Strobiomyces 118 S. strobilaceus S. cofusus S. seminudus S. seminudus Austroboletus 122 A. subvirens A. gracilis A. gracilis Boletellus 125 B. emodesis B. elatus B. russellii B. longicollis B. chrysenteroides Heimiella 130 H. japonica Russula 131 R. aurata R. sororia R. vesca R. cyanoxantha
5 Growth of ectomycorrhizal fungi on various culture media 169 Table 1. Derivation of the fungi examinedcontinue 135 R. mariae R. compacta R. emetica R. rubescens R. castanopsidis R. atropurpurea R. violeipes R. delica R. senecis R. nigricans R. foetens R. rosacea Lactarius 147 L. akahatsu L. chrysorrheus L. hatsudake L. hygrophoroides L. porninsis L. laeticolorus L. gerardii L. acris L. quietus L. indigo L. decipiens L. pterosporus L. controversus L. subzonarius L. mammosus L. ochrogalactus L. tabidus L. vellereus L. subplinthogalus L. glaucescens L. plumbeus Ramaria 168 R. botrytis Gomphus 169 G. fujisanensis Boletopsis 170 B. sp Albatrellus 171 A. confluens A. caeruleoporus Hymenogaster 173 H. arenarius Octavianina 174 O. asterosperma Rhizopogon 175 R. rubescens R. nigrescens R. superiorensis R. luteolus Hydnangium 179 H. carneum H. carneum Zelleromyces 181 Z. gikeyae Bulletin of FFPRI Vol.7 No
6 170 AKAMA K. et al. Table 1. Derivation of the fungi examinedcontinue Astraeus 182 A. hygrometricus A. hygrometricus A. hygrometricus var.koreanus Scleroderma 185 S. cepa S. areolatum S. bovista S. columnare S. reae S. reae S. citrinum S. citrinum S. verrucosum S. albidum S. macrorrhizom S. dictyosporum S. dictyosporum Pisolithus 198 P. tinctorius f.tinctorius P. tinctorius f.turgidus P. tinctorius f.pisocarpius Alpova 201 A. sp A. sp ) 生育温度 ml ml 90mm 40ml 4mm 1 NO Scheffe 3) 培地 8 Hagem Johnson, ( Yeast ) Difco PD 1976KO Melin-Norkrans Marx, 1969 MMN Johnson 1960 Ma/2 Abuzinadah 1986 MMa AK Table
7 Growth of ectomycorrhizal fungi on various culture media 171 Table 2 (g) Composition of the culture media used in the present study (Valus are grams in a liter of medium) Hagem Yeast PD KO MMN Ma/2 MMa AK (Difco) (Difco) (Difco) (Difco) 24 (1% ) 0.5(ml) 1.2(ml) ml 90mm ml 50ml Bulletin of FFPRI Vol.7 No
8 172 AKAMA K. et al. 1) 培養温度の影響 62 Table Table 3 Range of temperatures favorable for the growth of ectomycorrhizal fungi Cenococcum geophilum Suillus variegatus Armillaria albonaripes Alpova sp ) 培地の種類の影響 Table
9 Growth of ectomycorrhizal fungi on various culture media 173 Table 4. Growth of isolates of ectomycorrhizal fungi on eight different culture media. Values are means caluculaetd from three replicates. * mm Hagem Yeast PD KO MMN Ma/2 MMa AK mg DW mm mg DW mm mg DW Elaphomyces sp Elaphomyces sp Cenococcum geophilum Tuber borchii Armilaria albonaripes mm mg DW mm mg DW mm mg DW mm mg DW mm mg DW Bulletin of FFPRI Vol.7 No
10 174 AKAMA K. et al. Table 4. Growth of isolates of ectomycorrhizal fungi on eight different culture media. Values are means caluculaetd from three replicates.continue * mm Hagem Yeast PD KO MMN Ma/2 MMa AK mg DW mm mg DW mm mg DW mm mg DW mm mg DW mm mg DW mm mg DW mm mg DW
11 Growth of ectomycorrhizal fungi on various culture media 175 Table 4. Growth of isolates of ectomycorrhizal fungi on eight different culture media. Values are means caluculaetd from three replicates.continue * mm Hagem Yeast PD KO MMN Ma/2 MMa AK mg DW mm mg DW mm mg DW Suillus variegatus Gyroporus sp mm mg DW mm mg DW mm mg DW mm mg DW mm mg DW Bulletin of FFPRI Vol.7 No
12 176 AKAMA K. et al. Table 4. Growth of isolates of ectomycorrhizal fungi on eight different culture media. Values are means caluculaetd from three replicates.continue * mm Hagem Yeast PD KO MMN Ma/2 MMa AK mg DW mm mg DW mm mg DW Leccinum insigne mm mg DW mm mg DW mm mg DW mm mg DW mm mg DW
13 Growth of ectomycorrhizal fungi on various culture media 177 Table 4. Growth of isolates of ectomycorrhizal fungi on eight different culture media. Values are means caluculaetd from three replicates.continue * mm Hagem Yeast PD KO MMN Ma/2 MMa AK mg DW mm mg DW mm mg DW Lactarius decipiens L. mammosus L. plumbeus Boletopsis sp mm mg DW mm mg DW mm mg DW mm mg DW mm mg DW Bulletin of FFPRI Vol.7 No
14 178 AKAMA K. et al. Table 4. Growth of isolates of ectomycorrhizal fungi on eight different culture media. Values are means caluculaetd from three replicates.continue * mm Hagem Yeast PD KO MMN Ma/2 MMa AK mg DW mm mg DW mm mg DW Scleroderma columnare S. macrorrhizom Alpova sp Alpova sp mm mg DW mm mg DW mm mg DW mm mg DW mm mg DW
15 Growth of ectomycorrhizal fungi on various culture media LaccariaNo.1419 PD Hagem PD AK 2TricholomaNo.2026 Hagem PD AK 3AmanitaNo.2954 PD Yeast PD AK (Photo 1a) 5BoletusNo Yeast PD Ma/2 6TylopilusNo PD AK 7LeccinumNo Yeast PD Ma/2 PD AK 8RussulaNo LactariusNo PD PD 10SclerodermaNo Yeast PD AK Hagem MMN Photo 1. Growth of isolates of ectomycorrhizal fungi on agar media. AK AK MMa PD a, Amanita rubrovolvata on AK medium; b, Laccaria vinaceoavellanea on AK medium; c, Paxillus involutus on MMa medium; d, Suillus granulatus on PD medium. Fig 1 PD Hagem MMN 4SuillusNo.7180 Yeast PD Ma/2 AK (Photo 1d) Hagem KO MMN Bulletin of FFPRI Vol.7 No
16 180 AKAMA K. et al. 重量成長 (mg) Hagem 重量成長 (mg) Yeast 重量成長 (mg) PD 重量成長 (mg) KO 伸長成長 (mm) 伸長成長 (mm) 伸長成長 (mm) 伸長成長 (mm) 90 MMN 90 Ma/2 90 MMa 90 AK 重量成長 (mg) 重量成長 (mg) 重量成長 (mg) 重量成長 (mg) 伸長成長 (mm) 伸長成長 (mm) 伸長成長 (mm) 伸長成長 (mm) Fig.1. Relationship between extension growth and weight growth of ectomycorrhizal fungi Miller 1983 Brunderett 1996 Abuzinadah, R. A. and Read, D. J. (1986) The role of proteins in the nitrogen nutrition of ectomycorrhizal plants.. Utilization of peptides and proteins by ectomycorrhizal fungi, New Phytologist, 103, Brundrett, M., Bougher, N., Dell, B., Grove, T. and Malajczuk, N. (1996) Working with Mycorrhizas in Forestry and Agriculture, Australian Centre for International Agricultural Research, Canberra, 374p. Hacskaylo, E., Palmer, J. G. and Vozzo, J. A. (1965) Effect of temperature on growth and respiration of ectotrophic mycorrhizal fungi, Mycologia, 57, (1976) 17, Jonson, L. F., Curl, E. A., Bond, J. H. and Fribourg, H. A. (1960) Methods for studying soil microflora-plant disease relationships, Burgess Publishing Co., Minneapolis, 178p. Mark, D. H. (1969) The influence of ectotrophic mycorrhizal fungi on the resistance of pine roots to pathogenic infections.. Antagonism of mycorrhizal fungi to root pathogenic fungi and soil bacteria, Phytopathology, 59, Miller, O. K. Jr., Miller, S. L. and Palmer, J. G. (1983) Description and identification of selected mycorrhizal fungi in pure culture, Mycototaxon, 18, Molina, R., Massicotte, H. B. and Trappe, J. M. (1992) Specificity phenomena in mycorrhizal symbioses: Community-ecological consequences and practical implications. In Allen, M.F. (ed.) Mycorrhizal Functioning, Chapman & Hall, New York, ) Taniguchi, T., Kanazaki, N., Tamai, S., Yamanaka, N. and Futai, K. (2006) Does ectomycorrhizal fungal
17 Growth of ectomycorrhizal fungi on various culture media 181 community structure vary along a Japanese black pine (Pinus thunbergii) to black locust (Robinia pseudoavacia) gradient?, New Phytologist, 173, Taylor, J. B. (1971) A selective medium for the isolation of basidiomycetes from diseased roots, mycorrhizas, and soil, Trans. Br. mycol. Soc. 56, R D 902p. Bulletin of FFPRI Vol.7 No
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