WELCOME To my Thesis Presentation
Isolation of Trichoderma spp from different locationof Bangladesh and determination of their antifungal                   ...
OBJECTIVES            OBJECTIVES1. To   isolate   and   identify       theTrichoderma spp from the soil.2. To study thei...
MATERIALS AND METHODS          MATERIALS AND METHODS COLLECTION OF SOIL SAMPLE :   Soil sample was collected from differe...
Isolation of Trichoderma spp from soil  Isolation of Trichoderma spp from soilDilution plate technique (Nafiza, 2010) for ...
Photograph: Fungal colonies growing out of PDA media inoculated withsoil samples at 10-4 dilution
A                    B                             CD                    E                             F    Photograph . P...
A                                 B  C                                 DPhotograph . Pure culture of Trichoderma isolates....
Morphological characters of Trichoderma isolates  Mycelial growth  Colony characteristics        a) Colony shape        ...
Collection of soil borne plant pathogenThe test pathogen Penicillium sp, Fusarium oxysporum, Sclerotiumrolfsii, Rhizoctoni...
A                                    B                                  Fusarium oxysporum             A                  ...
A                                B                                          Rhizoctonia solani                     A      ...
A                                 B                                 Colletotrichum gloeosporiodes                  A      ...
Determination of antifungal properties of Trichoderma isolatesAntifungal effect of different Trichoderma isolates against ...
RESULTS
Morphological characters of TrichodermaMorphological characters of Trichoderma Isolates Name                   Mycelial   ...
Colony characteristics                         Colony characteristics      Isolates                    General characteris...
Spore density of Trichoderma isolatesSL      Trichoderma isolates     Sporulation capacityNO.                             ...
Effect of temperature on the growth of Trichoderma isolates
Determination of antifungal properties of Trichoderma Determination of antifungal properties of Trichodermaisolates isolat...
3 DAI                            6 DAI         A                                 BST6         A                           ...
Table. Antifungal activity of Trichoderma isolates against Fusariumoxysporium in dual culture                             ...
3 DAI                          6 DAI         A                           BST5         A                           BST8    ...
Table. Antifungal activity of Trichoderma isolates against Sclerotium rolfsii indual culture                              ...
3 DAI                       6 DAI       AA                          B BST5       A                           B BST8Photogr...
Table. Antifungal activity of Trichoderma isolates against Rhizoctonia solani indual culture                              ...
3 DAI                           6 DAI         A                              BST4         A                              B...
Table. Antifungal activity of Trichoderma isolates against Aspergillus flavus indual culture                              ...
3 DAI                             6 DAI         A                                BST5         A                           ...
Table. Antifungal activity of Trichoderma isolates against Colletotrichum  gloeosporiodes in dual culture                 ...
3 DAI                             6 DAI          A                               BST5          A                          ...
Table. Antifungal activity of Trichoderma isolates against Phomopsis vexans in  dual culture                              ...
3 DAI                            6 DAI        A                                 BST3         A                            ...
Comparison of antifungal activity the best three isolates (ST55,ST66Comparison of antifungal activity the best three isola...
Effect of Trichoderma isolates on the growth of different pathogenic fungi
SUMMARY AND CONCLUSION            SUMMARY AND CONCLUSIONCollected isolates were categorized into groups depending ontheir ...
Isolation of Trichoderma spp from different location of bangladesh and determination of their antifungal feature by Md. Ka...
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Isolation of Trichoderma spp from different location of bangladesh and determination of their antifungal feature by Md. Kamaruzzaman

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Bangladesh Agricultural university one of the leading Agricultural university of the South Asia. This University was situated in Mymensingh, Bangladesh. Department of Plant Pathology is a royal Department of this university.
In the year of 2012, an exclusive experiment was conducted in Department of Plant Pathology.

The Title was that Experiment Was: "Isolation of Trichoderma spp from different location of Bangladesh and determination of their antifungal Feature"

Abstract:
The research work was carried out to identify antifungal Trichoderma isolates from the rhizosphere to control soil borne plant pathogenic fungi. Ten Trichoderma isolates were isolated from the soil samples collected from different locations of Bangladesh. Their morphological characters and antifungal feature was studied. Colony diameter varied from 30.50 mm to 90.00 mm. The isolates were categorized into different groups based on their colony properties. On the basis of colony color, isolates were grouped into five such as dark green, green, light green, yellowish green and whitish green. Maximum isolates were green and light green. On the basis of growth habit and colony consistency the isolates were categorized into three groups, in which most species had fast growth and were compact in appearance. Different selected temperature such as: 20o, 25 o, 30o and 35 o C were maintained in incubators to observe the growth of different Trichoderma isolates. It was found that Trichoderma grew well at 25-30ºC. Antifungal activities of all Trichoderma isolates were tested against seven fungal mycoflora viz. Penicillium sp, Fusarium oxysporum, Sclerotium rolfsii, Rhizoctonia solani, Aspergillus flavus, Colletotrichum gloeosporiodes and Phomopsis vexans by duel culture method. All isolates of Trichoderma showed wide variation in growth inhibition against these pathogens. Among all of the Trichoderma isolates, ST5 was strongly inhibited the growth of all the pathogenic fungi.

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Isolation of Trichoderma spp from different location of bangladesh and determination of their antifungal feature by Md. Kamaruzzaman

  1. 1. WELCOME To my Thesis Presentation
  2. 2. Isolation of Trichoderma spp from different locationof Bangladesh and determination of their antifungal Feature MD. KAMARUZZAMAN ID No. : 11 Ag. P. Path JJ-07M Reg. No. : 33141 Session: 2006-2007 Phone: +8801722449614 Supervisor Co-SupervisorProf. Dr. M. Delwar Hossain Prof. Dr. Ismail HossainDepartment of Plant Pathology Department of Plant Pathology
  3. 3. OBJECTIVES OBJECTIVES1. To isolate and identify theTrichoderma spp from the soil.2. To study their wide range ofantifungal activity in invitro assay.
  4. 4. MATERIALS AND METHODS MATERIALS AND METHODS COLLECTION OF SOIL SAMPLE : Soil sample was collected from different location of Bangladesh such as Jamalpur, Sherpur, Netrokona, Tangail, Gazipur, Faridpur, Rangpur and BAU campus. Fig. Composite soil samples of different Agricultural fields
  5. 5. Isolation of Trichoderma spp from soil Isolation of Trichoderma spp from soilDilution plate technique (Nafiza, 2010) for isolation of Trichoderma sppwas followed: Photograph: Preparation of dilution series (Soil dilution)
  6. 6. Photograph: Fungal colonies growing out of PDA media inoculated withsoil samples at 10-4 dilution
  7. 7. A B CD E F Photograph . Pure culture of Trichoderma isolates. A: ST1 D: ST4 B: ST2 E: ST5 C: ST3 F: ST6
  8. 8. A B C DPhotograph . Pure culture of Trichoderma isolates. A: ST7 D: ST9 B: ST8 E: ST10
  9. 9. Morphological characters of Trichoderma isolates  Mycelial growth  Colony characteristics a) Colony shape b) Colony color c) Colony consistency d) Sporulation CapacityEffect of temperature on the growth of Trichoderma isolates Trichoderma isolates allowed growing at different selected temperature such as: 20o, 25o, 30o and 35oC on PDA media.
  10. 10. Collection of soil borne plant pathogenThe test pathogen Penicillium sp, Fusarium oxysporum, Sclerotiumrolfsii, Rhizoctonia solani, Aspergillus flavus, Colletotrichum sp andPhomopsis vexans were collected from IPM Laboratory,Bangladesh Agricultural University, Mymensingh; Plant DiseaseClinic, Department of Plant Pathology, Bangladesh AgriculturalUniversity, Mymensingh; Bangladesh Rice Research Institute,Gazipur. A B Penicillium sp Photograph: Different structures of isolated fungi, A) Pure culture, B) Different structures
  11. 11. A B Fusarium oxysporum A B Sclerotium rolfsiiPhotograph: Different structures of isolated fungi, A) Pure culture, B) Different structures
  12. 12. A B Rhizoctonia solani A B Aspergillus flavusPhotograph: Different structures of isolated fungi, A) Pure culture, B) Different structures
  13. 13. A B Colletotrichum gloeosporiodes A B Phomopsis vexansPhotograph: Different structures of isolated fungi, A) Pure culture, B) Different structures
  14. 14. Determination of antifungal properties of Trichoderma isolatesAntifungal effect of different Trichoderma isolates against Penicilliumsp, Fusarium oxysporum, Sclerotium rolfsii, Rhizoctonia solani,Aspergillus flavus, Colletotrichum gloeosporiodes and Phomopsisvexans were studied in dual culture method by Nafiza Rahman (2010).Percent inhibition of average radial growth was calculated following theformula of Edington et al., (1971).L = [(C – T)/C] x 100Where,L = inhibition of radial mycelial growth;C = radial growth measurement of the pathogen in control;T = radial growth of the pathogen in the presence of TrichodermaAnalysis of dataAnalysis of dataData collected during experimental period were tabulated andanalyzed through a standard computer package statistical procedureMSTAT-C (Gomez and Gomez, 1984).
  15. 15. RESULTS
  16. 16. Morphological characters of TrichodermaMorphological characters of Trichoderma Isolates Name Mycelial Mycelial diameter at 3 diameter at 6 DAI (mm) DAI (mm) ST1 86.62 d 90.00 a ST2 87.69 b 89.38 b ST3 62.58 e 86.69 c ST4 30.50 h 89.89 a ST5 89.67 a 90.00 a ST6 89.54 a 89.83 ab ST7 36.56 g 78.50 d ST8 89.39 a 90.00 a ST9 86.30 c 89.96 a ST10 44.00 f 85.86 c LSD 0.616 0.479
  17. 17. Colony characteristics Colony characteristics Isolates General characteristicsSL. Trichoderma Shape Color Growth ColonyNO. isolates habit consistency1. ST1 Regular Green Fast Compact2. ST2 Regular Whitish green Fast Loose3. ST3 Regular Dark green Medium Very Compact4. ST4 Regular Light green Slow Compact5. ST5 Regular Yellowish green Fast Compact6. ST6 Regular Whitish green Fast Compact7. ST7 Regular Whitish Green Slow Very Compact8. ST8 Regular Light green Fast Very Compact9. ST9 Regular Green Fast Loose10. ST10 Regular Dark green Medium Very Compact
  18. 18. Spore density of Trichoderma isolatesSL Trichoderma isolates Sporulation capacityNO. (Number of spore/ml) 1. ST1 0.84×1010 2. ST2 0.91×1010 3. ST3 0.84×1010 4. ST4 0.59×1010 5. ST5 1.10×1010 6. ST6 1.11×1010 7. ST7 0.93×1010 8. ST8 0.98×1010 9. ST9 0.74×1010 10. ST10 1.01×1010
  19. 19. Effect of temperature on the growth of Trichoderma isolates
  20. 20. Determination of antifungal properties of Trichoderma Determination of antifungal properties of Trichodermaisolates isolates Table. Antifungal activity of Trichoderma isolates against Penicillium sp in dual culture Penicillium sp 3 DAI 6 DAI Trichoderma Mycelial growth % Mycelial growth % isolates (mm) inhibition (mm) inhibition ST1 19.30 de 25.29 2.10 h 94.78 ST2 20.23 cd 20.44 7.26 e 81.96 ST3 20.40 c 19.78 8.16 d 79.73 ST4 22.16 b 12.84 6.36 f 84.20 ST5 18.23 f 28.31 4.43 g 89.00 ST6 10.40 h 59.10 0.46 i 98.84 ST7 20.30 cd 20.18 15.10 b 62.53 ST8 17.10 g 32.76 0.16 i 99.58 ST9 19.16 ef 24.63 10.23 c 74.60 ST10 21.20 bc 16.64 15.20 b 62.28
  21. 21. 3 DAI 6 DAI A BST6 A BST8 Photograph. Inhibition of mycelial growth of Penicillium sp by Trichoderma Isolates. A: Mycelial growth at 3DAI B: Mycelial growth at 6DAI
  22. 22. Table. Antifungal activity of Trichoderma isolates against Fusariumoxysporium in dual culture Fusarium oxysporium 3 DAI 6 DAITrichoderma Mycelial growth % Mycelial growth % isolates (mm) inhibition (mm) inhibition ST1 21.30 d 21.49 19.26 c 75.89 ST2 24.43 b 9.95 20.10 b 74.85 ST3 20.20 e 25.55 12.16 e 84.77 ST4 16.20 i 40.29 10.23 f 87.19 ST5 19.26 f 28.99 0.33 h 99.58 ST6 18.33 h 32.43 10.20 f 87.23 ST7 21.06 d 22.35 18.26 d 77.14 ST8 18.43 gh 32.06 9.23 g 88.44 ST9 19.20 fg 29.23 19.63 bc 75.43 ST10 23.10 c 14.86 20.13 b 74.81
  23. 23. 3 DAI 6 DAI A BST5 A BST8 Photograph. Inhibition of mycelial growth of Fusarium oxysporium by Trichoderma Isolates. A: Mycelial growth at 3DAI B: Mycelial growth at 6DAI
  24. 24. Table. Antifungal activity of Trichoderma isolates against Sclerotium rolfsii indual culture Sclerotium rolfsii 3 DAI 6 DAITrichoderma Mycelial growth % Mycelial growth % isolates (mm) inhibition (mm) inhibition ST1 26.36 f 63.97 20.26 f 77.52 ST2 30.26 d 58.65 26.20 e 70.94 ST3 26.06 f 64.38 4.23 i 95.30 ST4 29.13 e 60.20 19.33 g 78.55 ST5 21.16 h 71.08 0.26 j 99.70 ST6 33.33 c 54.46 30.13 c 83.12 ST7 33.23 c 54.59 36.50 b 59.61 ST8 23.10 g 68.44 15.50 h 95.33 ST9 33.16 c 54.68 15.20 h 83.14 ST10 34.26 b 53.18 28.13 d 89.85 Control 73.20 a 90.16 a LSD 0.60 0.82
  25. 25. 3 DAI 6 DAI AA B BST5 A B BST8Photograph. Inhibition of mycelial growth of Sclerotium rolfsii by Trichoderma Isolates. A: Mycelial growth at 3DAI B: Mycelial growth at 6DAI
  26. 26. Table. Antifungal activity of Trichoderma isolates against Rhizoctonia solani indual culture Rhizoctonia solani 3 DAI 6 DAITrichoderma Mycelial growth % Mycelial growth % isolates (mm) inhibition (mm) inhibition ST1 26.90 c 70.08 4.10 e 95.44 ST2 29.20 c 67.53 0.36 g 99.59 ST3 31.23 bc 65.27 14.30 d 84.11 ST4 29.40 c 67.30 0.23 g 99.74 ST5 27.16 c 69.79 2.10 f 97.66 ST6 26.16 g 70.90 0.13 g 99.85 ST7 36.20 b 59.74 22.23 b 75.29 ST8 16.13 d 82.00 0.40 g 99.55 ST9 32.56 bc 63.78 18.23 c 79.74 ST10 32.10 bc 64.30 0.13 g 99.85 Control 89.93 a 90.00 a LSD 6.44 0.75
  27. 27. 3 DAI 6 DAI A BST4 A BST6 Photograph. Inhibition of mycelial growth of Rhizoctonia solani by Trichoderma Isolates. A: Mycelial growth at 3DAI B: Mycelial growth at 6DAI
  28. 28. Table. Antifungal activity of Trichoderma isolates against Aspergillus flavus indual culture Aspergillus flavus 3 DAI 6 DAITrichoderma Mycelial growth % Mycelial growth % isolates (mm) inhibition (mm) inhibition ST1 23.10 de 42.53 12.23 d 80.01 ST2 24.26 cd 39.63 6.30 g 89.70 ST3 20.13 ef 49.91 9.26 e 84.85 ST4 26.23 bc 34.74 5.30 h 91.33 ST5 24.56 cd 38.88 7.20 f 88.23 ST6 19.16 f 52.23 0.00 i 100 ST7 28.30 b 29.60 22.30 c 63.56 ST8 23.20 d 42.28 0.00 i 100 ST9 28.23 d 29.76 23.46 b 61.65 ST10 24.26 cd 39.63 23.10 b 62.25 Control 40.20 a 61.20 a LSD 2.97 0.74
  29. 29. 3 DAI 6 DAI A BST5 A BST8 Photograph. Inhibition of mycelial growth of Aspergillus flavus by Trichoderma Isolates. A: Mycelial growth at 3DAI B: Mycelial growth at 6DAI
  30. 30. Table. Antifungal activity of Trichoderma isolates against Colletotrichum gloeosporiodes in dual culture Colletotrichum gloeosporiodes 3 DAI 6 DAITrichoderma Mycelial % Mycelial growth % isolates growth inhibition (mm) inhibition (mm) ST1 26.10 b 50.06 33.10 b 55.92 ST2 20.46 d 60.84 12.16 h 83.79 ST3 18.36 e 64.85 13.36 g 82.20 ST4 19.43 d 62.81 23.46 c 68.75 ST5 18.26 e 65.05 6.10 i 91.87 ST6 19.53 d 62.62 4.13 j 91.49 ST7 17.36 e 66.77 15.56 f 79.27 ST8 23.10 c 55.80 18.30 e 75.63 ST9 23.60 c 54.84 19.26 d 74.34 ST10 27.13 b 48.08 12.26 h 83.66
  31. 31. 3 DAI 6 DAI A BST5 A BST8 Photograph. Inhibition of mycelial growth of Colletotrichum gloeosporiodes by Trichoderma Isolates. A: Mycelial growth at 3DAI B: Mycelial growth at 6DAI
  32. 32. Table. Antifungal activity of Trichoderma isolates against Phomopsis vexans in dual culture Phomopsis vexans 3 DAI 6 DAITrichoderma Mycelial % Mycelial growth % isolates growth Inhibition (mm) inhibition (mm) ST1 11.16 f 82.3 4.33 f 94.23 ST2 14.10 c 77.71 8.23 d 89.06 ST3 12.26 e 80.01 0.00 g 100 ST4 10.26 g 83.77 20.20 c 73.16 ST5 10.13 g 83.98 0.50 g 99.33 ST6 9.10 h 85.61 0.13 g 99.82 ST7 18.20 b 71.23 21.33 b 71.65 ST8 13.16 d 79.18 9.23 d 87.73 ST9 13.033 d 79.40 4.16 f 94.46 ST10 11.33 f 82.08 6.33 e 91.58
  33. 33. 3 DAI 6 DAI A BST3 A BST6 Photograph. Inhibition of mycelial growth of Phomopsis vexans by Trichoderma Isolates. A: Mycelial growth at 3DAI B: Mycelial growth at 6DAI
  34. 34. Comparison of antifungal activity the best three isolates (ST55,ST66Comparison of antifungal activity the best three isolates (ST , STand ST88)and ST ) Pathogens culture Mycelial growth (mm) at 6 DAI ST5 ST6 ST8 Penicillium sp 4.43 0.46 0.16 Fusarium oxysporium 0.33 10.20 9.23 Sclerotium rolfsii 0.26 15.22 4.20 Rhizoctonia solani 2.10 0.13 0.40 Aspergillus flavus 7.20 0.00 0.00 Colletotrichum sp 6.10 4.13 18.30 Phomopsis vexans 0.50 0.13 9.23 Table. Antifungal effect of selected three Trichoderma isolates
  35. 35. Effect of Trichoderma isolates on the growth of different pathogenic fungi
  36. 36. SUMMARY AND CONCLUSION SUMMARY AND CONCLUSIONCollected isolates were categorized into groups depending ontheir cultural, morphological and molecular characterizationslike colony color, mycelial growth and pattern.The isolates were classified into 5 categories depending on color,which were dark green, green, light green, yellowish green andwhitish green color.It was observed that Trichoderma grow well at 25-30oC.Antifungal activity of Trichoderma isolates against sevenpathogenic fungi was studied by duel culture method.In this experiment, we found that ST5 strongly inhibit the growthof all tested pathogenic fungi.

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