Activity of laccase  and the fruiting body formation of  Pleurotus spp . Presented By Mr. Suparat Laohapitakvorn Mentor Dr...
<ul><li>Most mushroom which are cultivated in sawdust do not yield desirable amount of produce. This become major problem ...
Introduction <ul><li>Oyster Mushroom -  Pleurotus ostreatus  </li></ul>Bhutanese oyster mushroom  -  Pleurotus  sajor-caju
<ul><ul><li>Edible mushrooms are always grown in sawdust with  </li></ul></ul><ul><ul><li>supplements . </li></ul></ul>
<ul><li>Be able to decompose lignocellulose  </li></ul><ul><li>Enzymes involved in  lignocellulolytic  degradation are  ce...
Objective <ul><li>To study relationship between laccase activities and fruiting body production </li></ul>
Mushroom culture <ul><li>stock culture  in   PDA </li></ul><ul><li>In  Potato Dextrose Agar (PDA)  </li></ul><ul><li>Asept...
Measure mycelial growth in PDA <ul><li>Grow in PDA each  5  replication </li></ul><ul><li>Incubate at room temperature for...
Laccase assay in PDA <ul><li>3  replication of each mushroom culture on PDA for 4 days at room temperature. </li></ul><ul>...
Cultivation in PDB <ul><li>Potato Dextrose Broth (PDB) </li></ul><ul><li>10  pieces of inoculums/flask, 4replications/stra...
 
Culture in sawdust bag <ul><li>Fully grow mushroom culture in sorghum seed (2 weeks) </li></ul><ul><li>Inoculate to sawdus...
 
 
Result : 1 <ul><li>Measurement of mycelial growth in PDA   </li></ul><ul><li>Pleurotus sajor-caju  (No. 1-3) showed  dense...
1
Result : 2 + mean dark +++  all area of colony 4 +++  center of colony 3 +  all area of colony 2 ++++  center of colony 1 ...
 
Result : 3 <ul><li>The activity of No. 1, 2 and 3 increased daily in day 4, 6 and 8 </li></ul><ul><li>The activity of No.4...
 
Laccase acting in sawdust bag <ul><li>specific activity =   laccase activity (Unit) </li></ul><ul><ul><ul><li>protein cont...
 
Mushroom Production <ul><li>The data of weight of each strain and amount of fruiting body was record for 2 months </li></u...
Pleurotus sajor-caju  Number  1 Pleurotus ostreatus  Number  4
Production Mushroom 27.87 96.12 1.52 1672 5767 91 60 4 20.25 121.64 3.09 1134 6812 173 56 3 22.71 124.58 3.15 1340 7350 18...
Conclusions <ul><li>Correlation was found between laccase in  PDB ( 8 th  day mycelia )  and total fruiting body productio...
Flush / bag Total weight (g)/bag In PDB
 
Discussions <ul><li>Laccase activity (in PDB) and sawdust correlates with number of flushing bag, total weight and total a...
Acknowledgement <ul><li>Junior Science Talent Project </li></ul><ul><li>Dr. Charida Pukahuta </li></ul><ul><li>Assist. Pro...
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mushroom

  1. 1. Activity of laccase and the fruiting body formation of Pleurotus spp . Presented By Mr. Suparat Laohapitakvorn Mentor Dr. Charida Pukahuta Assist. Prof. Dr. Weerasuk Suraruengchai
  2. 2. <ul><li>Most mushroom which are cultivated in sawdust do not yield desirable amount of produce. This become major problem for mushroom farmers . </li></ul>Why I did project ?
  3. 3. Introduction <ul><li>Oyster Mushroom - Pleurotus ostreatus </li></ul>Bhutanese oyster mushroom - Pleurotus sajor-caju
  4. 4. <ul><ul><li>Edible mushrooms are always grown in sawdust with </li></ul></ul><ul><ul><li>supplements . </li></ul></ul>
  5. 5. <ul><li>Be able to decompose lignocellulose </li></ul><ul><li>Enzymes involved in lignocellulolytic degradation are cellul a se, hemi-cellulase, xylanase, phenoloxida ses example laccase </li></ul><ul><li>Laccase relates in </li></ul><ul><ul><li>Decomposition of lignocellulose </li></ul></ul><ul><ul><li>Stage of fruiting body formation </li></ul></ul><ul><ul><li>Be able to detoxify organic wastwater </li></ul></ul>
  6. 6. Objective <ul><li>To study relationship between laccase activities and fruiting body production </li></ul>
  7. 7. Mushroom culture <ul><li>stock culture in PDA </li></ul><ul><li>In Potato Dextrose Agar (PDA) </li></ul><ul><li>Aseptic technique </li></ul><ul><li>incubate at room temperature for 7 days </li></ul>
  8. 8. Measure mycelial growth in PDA <ul><li>Grow in PDA each 5 replication </li></ul><ul><li>Incubate at room temperature for 9 days </li></ul><ul><li>Measure colony diameter by x-axis and y-axis daily </li></ul><ul><li>Record and calculate average growth of each culture </li></ul>
  9. 9. Laccase assay in PDA <ul><li>3 replication of each mushroom culture on PDA for 4 days at room temperature. </li></ul><ul><li>Assay with o-tolidine solution by pouring on mushroom mycelia. </li></ul><ul><li>Observe the color changes of solution to dark blue and record data. </li></ul>
  10. 10. Cultivation in PDB <ul><li>Potato Dextrose Broth (PDB) </li></ul><ul><li>10 pieces of inoculums/flask, 4replications/strain </li></ul><ul><li>Incubation 150 rpm., 26 o C </li></ul><ul><li>Laccase assay at day 4, 6 and 8 by measurement of color changes of o-tolidine and PDB, using Spectrophotomiter at 590 nm. </li></ul>
  11. 12. Culture in sawdust bag <ul><li>Fully grow mushroom culture in sorghum seed (2 weeks) </li></ul><ul><li>Inoculate to sawdust with supplements in plastic bags. Incubate at room temperature (1 month) </li></ul><ul><li>Move to fruiting house </li></ul><ul><li>Watering daily </li></ul><ul><li>Harvest the fruiting body and record the weight. </li></ul>
  12. 15. Result : 1 <ul><li>Measurement of mycelial growth in PDA </li></ul><ul><li>Pleurotus sajor-caju (No. 1-3) showed dense mycelia in horizontal growth (substrate mycelia) and vertical growth (aerial mycelia) </li></ul><ul><li>Pleurotus ostreatus (No. 4) showed dense mycelia in vertical growth </li></ul><ul><li>Pleurotus sajor-caju Number 1-3 grew about 12 – 13 mm/day but Pleurotus ostreatus Number 4 grew 9 mm/day </li></ul>
  13. 16. 1
  14. 17. Result : 2 + mean dark +++ all area of colony 4 +++ center of colony 3 + all area of colony 2 ++++ center of colony 1 Color change ( 3 replications) Number
  15. 19. Result : 3 <ul><li>The activity of No. 1, 2 and 3 increased daily in day 4, 6 and 8 </li></ul><ul><li>The activity of No.4 increased in day 4 and 6 but decreased in day 8 </li></ul>
  16. 21. Laccase acting in sawdust bag <ul><li>specific activity = laccase activity (Unit) </li></ul><ul><ul><ul><li>protein content ( Lowry method) </li></ul></ul></ul>
  17. 23. Mushroom Production <ul><li>The data of weight of each strain and amount of fruiting body was record for 2 months </li></ul><ul><li>The weight of production was high during day 16-30 </li></ul><ul><li>The weight of production was stable during day 48-62 </li></ul>
  18. 24. Pleurotus sajor-caju Number 1 Pleurotus ostreatus Number 4
  19. 25. Production Mushroom 27.87 96.12 1.52 1672 5767 91 60 4 20.25 121.64 3.09 1134 6812 173 56 3 22.71 124.58 3.15 1340 7350 186 59 2 13.11 126.08 3.17 826 7943 200 63 1 Amou nt of fruiting body Total Weig ht (g) Flu sh Amount of fruiting body Total weight (g) Flu sh Average/bag Total production Am Ou -nt of Bags Mush room No.
  20. 26. Conclusions <ul><li>Correlation was found between laccase in PDB ( 8 th day mycelia ) and total fruiting body production. </li></ul><ul><li>The mushroom No.1,2 and 3 had high laccase activity and high fruiting body production. </li></ul><ul><li>The mushroom No. 4 had lower laccase activity and lower fruiting body production than the others. </li></ul>
  21. 27. Flush / bag Total weight (g)/bag In PDB
  22. 29. Discussions <ul><li>Laccase activity (in PDB) and sawdust correlates with number of flushing bag, total weight and total amount of fruiting body </li></ul><ul><li>The kit to test the fruiting body may be invented by using this correlation </li></ul>
  23. 30. Acknowledgement <ul><li>Junior Science Talent Project </li></ul><ul><li>Dr. Charida Pukahuta </li></ul><ul><li>Assist. Prof. Dr. Weerasuk Suraruengchai </li></ul><ul><li>Teachers from Mahidol Wittayanusorn School </li></ul>

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