Heterobasidion insulare

Transcription

1 17(:31-39, ,4) 2,3) ph ph g/l 60 g/l 240 g/l N tyrosine cysteine N 9 B1 thiamine-hcl inositol, C ascorbic cacid pantothenic acid ( Research paper Root and Butt Rot of Pinus luchuensis Caused by and Its Nutrient Physiology Pi-Chao Yen, 1,4) Tun-Tschu Chang, 2,3) Shing-Rong Kuo Summary Department of Forestry, National Taiwan University. 1 Roosevelt Rd., Sec. 4, Taipei 106, Taiwan Division of Forest Protection, Taiwan Forestry Research Institute, 53 Nanhai Rd., Taipei 100, Taiwan. 3) Corresponding author, ttchang@serv.tfri.gov.tw 4) Part of data from MS thesis of the first author Received June 2001, Accepted October 2001.

2 32 was first observed on dead butt and root of Pinus luchuensis at Shihding, Taipei County in Later, it was commonly found on plantations of P. luchuensis in northern Taiwan. The white mycelial mat was found between the bark and wood of decayed butts and roots. Seedlings of 4 pine species, P. luchuensis, P. thunbergii, P. morrisonicola, and P. taiwanensis as well as Taiwania cryptomerioides, were inoculated with grown on pine twig fragments or sawdust medium. After 7 wk, some of the inoculated seedlings of P. luchuensis and P. thunbergii had died, and the fungus was reisolated from the diseased pines. The other 3 tree species remained healthy during the inoculation experiments. This is the first report of in Taiwan. The mycelial growth occurred in a temperature range of from 12 to 32, and the optimum temperature was 24. The optimum ph for mycelium growth of the fungus was at , and the best ph was at 5.2. Pectin, glucose, galactose, and mannitol were good carbon sources for mycelial growth. Optimum concentrations of glucose for mycelial growth were at g/l, while increasing glucose concentrations inhibited growth, and it stopped at a concentration of 240 g/l. Ammonium chloride and calcium nitrate were the best inorganic nitrogen sources for mycelial growth. Most amino acids were favorable to mycelial growth, while tyrosine and cysteine were not favorable. The optimum nitrogen normalities of ammonium chloride for mycelial growth were at N, whereas mycelial growth was in proportion to the concentration of asparatic acid in the range of N of nitrogen. Thiamine-HC1 one of the 9 vitamins tested, had the greatest enhancing effect on mycelial growth. Moreover, pantothenic acid, ascorbic acid, and inositol also enhanced growth of mycelium. Key words, Pinus luchuensis, root and butt rot, nutrient physiology. Yen PC, Chang TT, Kuo SR Root and butt rot of Pinus luchuensis caused by Heterobasidion insulare and its nutrient physiology. Taiwan J For Sci 17(: Heterobasidion H. annosum, H. auraucariae H. insulare (Buchanan 1988) H. auraucariae H. annosum H. auraucariae H. annosum (Buchanan 1988) Heterobasidion (Chang 1999) 1995 (Pinus luchuensis) H. insulare mm MA 2 malt-extract, 2 agar PDA Bacto potato-dextroseagar MA PDA

3 17(:31-39, ( Pinus morrisonicola) (P. taiwanensis) (P. thunbergii) (P. luchuensis) (Taiwania cryptomerioides) cm 15 cm ml 150 ml cm (Table parafilm 3-5 (P. taiwanensis) 1 cm 3 cm 30 min 300 ml 150 ml 3 10 g parafilm H. insulare 24 3 Table 1. Pathogenicity of on its hosts Plant species inoculated Disease incidence Exp. I Exp. II Inoculation of twigs Pinus thunbergii 50 (5/10) 18 (2/1 Pinus luchuensis 15.4 (2/13) 30.8 (4/13) Pinus morrisonicola 0 (0/1 0 (0/8) Pinus taiwanensis 0 (0/1 0 (0/1 Taiwnnia cryptomerioides 0 (0/8) 0 (0/10) Inoculation of sawdust Pinus thunbergii 22.2 (2/9) 30 (3/10) Pinus luchuensis 40 (2/5) 33.3 (4/1 Pinus morrisonicola 0 (0/10) 0 (0/1 Pinus taiwanensis 0 (0/9) 0 (0/13) Taiwania cryptomerioides 0 (0/10) 0 (0/1 In parentheses number of trees killed by the pathogen / number of trees inoculated with the pathogen. This data were recorded 1 yr after inoculation. MA ( 9 cm) 300 ml 50 ml (1 ) h 0.3 mm PDA MA 12, 16, 20, 24, 28, g KH 2 PO g Ca (NO 3 ) 2 4H 2 O 0.5 g MgSO 4 7H 2 O 30 g glucose 2 g Bacto malt extract 100 ml

4 34 1 N NaOH HCl ph ph Jensen's medium (Tuite 1969) 0.8 g KH 2 PO g MgSO 4 7H 2 O 1.5 g NH 4 NO ml 1 ppm thiamine-hc1 40 g 1000 ml Jensen's medium 1 N NaOH HCl ph mannitol, myo-inositol sorbitol 6 glucose, galactose, mannose, arabinose, xylose fructose 5 saccharose, maltose, lactose, cellulobiose trehalose 4 dextrin, soluble starch, pectin cellulose powder 1000 ml g 500 ml 150 ml L-form alanine, arginine, asparagines, asparatic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine valine asparatic acid N 0.8 g KH 2 PO g MgSO 4 7H 2 O 40 g glucose 2.66 g asparatic acid 1000mL 9 B1 thiamine-hcl B2 riboflavin B6 pyridoxine-hcl pantothenic acid H biotin inositol B12 vitamine B12 nicotinic acid C ascorbic acid ppm 3 40 g/l N NH 4 NO 3 NaNO 3 KNO 3 Ca(NO 3 ) 2 4H 2 O NH 4 Cl NaNO 2 NH 4 Cl N (Fig N 20

5 17(:31-39, (Fig. 1 (downy) (farinaceous) (cottony) (pellicular) (Fig. 3) 9 7 2cm mm 2-4 (Fig. 4) 8.5 m 8-20 m m (dextrinoid) m 3.0 m m lactase m peroxidase tyrosinase PDA MA mm 5 Fig. 1. White mycelial mat between the bark and wood of decayed butts and roots. Fig. 3. Colony of on PDA (left) and MA (right) media. Fig. 2. Basidioma of. Fig. 4. Conidiophores and conidia of (Spiniger sp.) produced on PDA and MA media.

6 36 5 (Table PDA MA (Fig. 5) ph ph 6.0 ph 1.86 (Fig. 6) (pectin) (dextrin) (glucose) (galactose) (mannitol) (Table Fig. 5. Growth rate of on PDA and MA media at indicated temperatures. (Fig. 7) g/l 40 g/l 40 g/l 240 g/l (Table 3) 0.02 N 40 g/l Fig. 6. Growth rate of at indicated ph values. 1 N (Fig. 8)

7 17(:31-39, Table 2. Effects of carbon sources on the growth of Carbon source Mycelium dry weight (mg) Alcoholic sugar Mannitol 31.5 b Myo-inositol 8.1 e Sorbitol 24.6 c Monosaccharide Glucose 30.8 b Galactose 29.5 b Mannose 30.9 b Arabinose 10.1 e Xylose 20.6 c Fructose 13.8 d Oligosaccharide Saccharose 29.4 b Maltose 12.3 d Lactose 8.6 e Cellulobiose 8.8 e Trehalose 13.7 d Polysaccharide Dextrin 29.1 b Starch soluble 13.2 d Pectin 66.3 a Cellulose powder 9.1 e After growing in liquid media at 24 for 14 d. Means followed by the same letter do not significantly differ at P = 0.05 according to Duncan`s multiple range test. (tyrosine) acid) (Table 4) (cysteine) (asparatic 40 g/l N (Fig. 9) Fig. 7. Growth rate of at indicated concentrations of glucose. Table 3. Effects of inorganic nitrogen sources on the growth of Inorganic nitrogen Mycelium dry weight (mg) source (0.375 N) NH 4 NO c NH 4 Cl 61.4 a Na NO b K NO b Ca (NO 3 ) a Na NO 2 0e Control 3) 11.2 d After growing in liquid media at 24 for 14 d. Means followed by the same letter do not significantly differ at P = 0.05 according to Duncan`s multiple range test. 3) No nitrogen source was used as the control. Fig. 8. Growth rate of at indicated concentrations of NH 4 NO 3. Y = 35.0X R 2 = 0.94 Fig. 9. Growth rate of at indicated concentrations of asparatic acid.

8 38 Table 4. Effects of amino acids on the growth of Amino acid Mycelium dry weight (mg) Asparatic acid 73.5 a Arginine 71.6 a Glutamine 60.9 a Asparagine 59.6 a Glutamic acid 64.2 a Lysine 68.3 a Valine 51.6 b Phenylalanine 40.2 cd Alanine 61.2 a Glycine 52.2 ab Serine 55.1 ab Proline 50.8 ab Methionine 47.7 bc Leucine 40.9 cd Isoleucine 48.1 bc Threonine 46.8 bc Histidine 47.5 bc Tryptophan 35.2 cd Tyrosine 8.3 e Cysteine 5.8 E Control 3) 60.2 a After growing in liquid media at 24 for 14 d. Means followed by the same letter do not significantly differ at P = 0.05 according to Duncan`s multiple range test. 3) NH 4 Cl was added as the nitrogen source. Table 5. Effects of growth factors on the growth of Growth factor Concentration (ppm) Thiamine-HCl 68.5 c 90.4 b a Riboflavin 70.3 a 72.8 a 60.3 b Pyridoxine-HCl 50.8 b 60.3 a 38.8 c Pantothenic acid 53.8 b 48.5 b a Biotin 49.2 b 46.1 b 61.7 a Inositol 71.4 b 99.2 a 73.6 b Vitamin B b 51.5 b 54.2 a Nicotinic acid 54.6 b 68.6 a 49.1 c Ascorbic acid 63.8 b 68.9 c a Mycelial dry weight (mg) was measured after growing in liquid media at 24 for 14 d. Means followed by the same letter for the same growth factor do not significantly differ at P = 0.05 according to Duncan`s multiple range test. thiamine-hcl inositol ascorbic acid pantothenic acid thiamine-hcl ascorbic acid pantothenic acid 50 ppm inositol 10 ppm (Table 5) H. insulare H. insulare 1995 ph (Highley and Kirk 1979) (Highley and Kirk 1979) ph (Highley and Kirk 1979)

9 17(:31-39, (Levi et al. 1968) (Highley and Kirk 1979) tyrosine cysteine 0.2N (Chang and Chen 1984) thiamine-hcl thiamine-hcl (Highley and Kirk 1979) thiamine- HCl inositol ascorbic acid pantothenic acid 40 g glucose 2.66 g asparatic acid 0.8 g KH 2 PO g MgSO 4 7H 2 O 1 ppm thiamine-hcl 1000 ml 1 N HCl 1 N NaOH ph 5.20 Buchanan PK A new species of Heterobasidion (Polyporaceae) from Australasia. Mycotaxon 32: Chang TT (ed.) List of the fungi in Taiwan. Council of Agriculture, Republic of China. 289 p. Chang TT, Chen T Study on nutrient physiology of three subgenus Ganoderma spp. in Taiwan. Phytopathol Entomol (National Taiwan Univ) 11: [in Chinese with English summary]. Highly TL, Kirk TK Mechanisms of wood decay and unique features of heart rot. Phytopathology 69: Levi MP, Merrill W, Cowling EB Role of nitrogen in wood deteriation VI. mycelial fractions and model nitrogon compound as substrances for growth of Polyporus sicolor and other wood-destroying and wood-inhibiting fungi. Phytopathology 58: Tuite J Plant pathological methods : fungi and bacteria. Lafayette, Indiana, USA. 39 p.