12 1 97 11 30 ( ) E-Mail wcho@mail.tajen.edu.tw 80 1000 26 (Trichoderma spp.) T20 T21 24 T21 T22 T26 46 63 33 19 600 100 800 800 1000 500 200 2000 Abstract In this study, chemical pesticide, antimicroorganisms Chinese medicines and Trichoderma spp. were used to inhibition the root rot disease of angelica. In chemical pesticide test, one of four pesticide, 80% Metirum was the best pesticide chemical, when the chemical dilution to 1000-fold, the spores still were inhibitory. In antagonistic test, Trichoderma strain T20 and T21 both were inhibitory the mycelium growth of F. solani. In the antibiotic test, T21, T22 and T26 strains were produce antibiotic, the percentage of inhibitory rate were 46, 63, and 33%. The Chinese medicine test showed, the gallnut and myrobalan were the best effect of inhibitory to F. solani in 19 Chinese medicines. When gallnut water extract dilute to 600-fold and myrobalan water extract dilute to 100-fold, could inhibition the spores germination of pathogenic fungi. When the water extract of gallnut in 800-fold, the myrobalan also could inhibitory the pathogen in 800-fold. When gallnut in 1000-fold, the inhibitory rate was 100% of mycobalance in 500-fold. When mycobalance in 200-fold, the 100% 1
inhibitory rate on gallnut was 2000-fold. Keyword Plant source pesticide, root rot disease of angelica, Trichoderma, anti-microorganism Chinese medicine (Angelica spp. L.) (Apiaceae) ( 1999 2003) A. sinensis A. gigas 50 ( 2009) ITS 1 ITS 2 5S 5.8S rdna (A. acutiloba) Hong et al., 2009 ( 2008) 2008 Fusarium spp. Fusarium spp. R3 R4 R5 4 12 500 R4 7 100 100 0 100 1000 ( 1) 26 1/2PSA T4 T20 T21 T22 T25 T26 T27 7 3 15 6 2
T20 T21 24 26 T21 ( 2) 26 1/2PSA 72 T21 T22 T26 3 R4 1/2PSA 46 63 33 T22 ( 1) 19 7 10 0 100 ( 4) 600 100 1 ( 5) 1000 500 6 500 600 100 600 100 1200 200 10 8 T20 10 8 T21 10 8 T20 T21 600 33.3 600 100 41.7 ( 2) 1. 2. (PCR) Fusarium solani 3. F. solani (Martius) Snyder & Hansen. 4. 24 32 28 5. T21 T22 6. 600 3
4 100 600 2005 77 2005 62 2002 365 ( ) 2008 97 179 2009 Rhizoctonia solani 98 61 2006 Alternaria brassiccola 67 2000 72 2001 91 2006 (Phytophthora capsici) 78 2002 50
1 Inhibition effect of fungicides Conidia germination (%) at n fold dilution R3 R4 R5 Prochloraz( ) 500X 10 7 11 1000X 36 40 49 1500X 76 83 82 2000X 100 100 100 Prochlorate manganese( ) 500X 100 100 100 1000X 1500X 2000X Carbendazim( ) 500X 100 100 100 1000X 1500X 2000X Metiram ( ) 500X 0 0 0 1000X 0 0 0 1500X 41 100 60 2000X 100 100 100 Check 100 100 100 1 Germination were tested on double and Tenfold cavity slides. Three replicates were corded for each treatment. The experiment was test for twice. 5
2 R4 Table 2. The antagonistic effect of Trichoderma isolates on the linear growth of Fusarium solani R4 isolate Isolates Linear growth(mm) Linear growth of R4 1 Growth inhibition rate(%) Antagonistic ability of Trichoderma isolates coverage entwine superparasitis T1 32.3 1.2 9 T2 32.3 1.5 9 T4 30.0 0.0 16 T5 31.0 0.0 13 T6 30.7 0.6 14 T7 31.7 0.6 11 T8 35.7 0.6 0 T9 33.3 0.6 7 T10 31.3 2.1 12 T11 35.7 0.6 0 T12 35.7 0.6 0 T13 33.0 0.0 7 T14 33.0 1.0 7 T15 32.0 1.0 10 T16 35.3 0.6 1 T17 30.7 0.6 14 T18 32.3 0.6 9 T19 32.0 1.0 10 T20 27.0 1.0 24 T21 26.3 0.6 26 T22 29.7 0.6 17 T23 31.7 0.6 11 T24 31.0 1.0 13 T25 29.7 0.6 17 T26 29.0 1.0 19 T27 30.3 0.6 15 Check 35.7 0.6 0 Mycelial discs of R4 (6mm in diameter) were inoculated on the center of the 1/2PSA plates and the mycelial discs (6mm in diameter) of Trichoderma isolates were inoculated on the two margins of the plates. Three plates were used for each isolate and all plates were incubated at 28. The linear growths of R4 were measured 3 days after inocubation. Growth inhibition rate = 100% ((Trichoderma isolate/ck) 100%). R4 colony was overed/entwined/superparasitism by Trichoderma isolates 3 days after incubation. R4 colony was not be covered/entwined/superparasitism by Trichoderma isolates 3 days after incubation. Three replicates were corded for each treatment. The experiment was test for twice. m 6
Mycelial linear growth (mm) Trichoderma 1 isolates Fig 1. The anti fungi mycelial growth effect of Trichoderma isolates to Fusarium solani R4 isolate. 7
3 10 R4 Table 3. Inhibition effect of 10 fold dilution aqueous extracts of nineteen Chinese to germination of Fusarium solani Scientific name Germination (%) 1 1. Myrobalan ( ) 0 2. Clove ( ) 0 3. Sehisandra chinensis ( ) 0 4. Cinnamon ( ) 0 5. Knotweed ( ) 0 6. Burnet bloodwort ( ) 0 7. Baical skullcap ( ) 0 8. Galla rhois ( ) 0 9. Anemarrhena ( ) 100 10. Rhei Rhizoma ( ) 100 11. Angelica ( ) 100 12. Coptis ( ) 100 13. Chinese senega ( ) 100 14. Foetid cassia ( ) 100 15. Chinese dittany ( ) 100 16. Amur cork tree ( ) 100 17. Wolfberry ( ) 100 18. Fleeceflower ( ) 100 19. Notoginseng ( ) 100 20.Water (Check) 100 Three replicates were used for each herb and 100 spores were counted for each replicate. Percentage of germination was based on the average of two tests. 8
4 Table 4. Inhibition effect of different dilution rate of eight antifungi Chinese herbs aqueous extracts to conidia germination of Fusarium solani Source of extract Conidia germination (%) at n fold dilution 1 50 100 200 400 600 700 Galla rhois ( ) 0 0 0 0 0 100 Myrobalan ( ) 0 1 100 Clove ( ) 0 100 Baical skullcap ( ) 0 100 Sehisandra chinensis ( ) 6 100 Knotweed ( ) 48 100 Cinnamon ( ) 97 100 Burnet bloodwort ( ) 94 100 Water (control) 100 1 Germination were tested on double and Tenfold cavity slides. Three replicates were corded for each treatment. The experiment was test for twice. 9
R4 Table 5. Aqueous extracts of Chinese herbs of compound from first selection on germination of conidia of Fusarium solani at various dilutions Chinese herbs of compound Dilution rates of Myrobalan aqueous extracts 1 Germination were tested on double and Tenfold cavity slides. Three replicates were corded for each treatment. The experiment was test for twice. Conidia germination (%) Dilution rates of Galla rhois aqueous extracts 600 800 1000 1200 1400 1600 1800 2000 Check 100 0 0 0 0 0 0 0 0 0 200 0 0 0 0 0 0 0 0 100 300 0 0 0 36 100 100 100 100 100 400 0 0 0 100 100 500 0 0 0 100 100 600 0 0 100 100 700 0 0 100 100 800 0 0 100 100 900 0 43 100 100 1000 0 83 100 100 Check 0 100 100 100 100 100 100 100 100 10
2 Fig 2. Control of Angelica root rot by antagonistic microorganisms and Chinese herbal medicines. 11