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TISSUE CULTURE ON BANANA SUCKERS AND RICE CALLUS (MICROPROPAGATION)

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UNIVERSITI MALAYSIA KELANTAN
UNIVERSITI MALAYSIA KELANTAN

This document summarizes a student group's plant tissue culture project on banana propagation. It includes: 1) The group members and their lecturer. The objective is to understand plant tissue culture procedures. 2) Banana suckers are commonly used as explants in tissue culture. The document describes sterilization and preparation of MS media for culturing banana shoot tips. 3) The results showed both contaminated and uncontaminated cultures. Factors affecting contamination and growth are discussed. Suggestions for improvement include using meristematic shoot tips and adding antioxidants to the media.

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PLANT TISSUE CULTURE MEMBERS : KAREN LENGGIAU ANK AUGUSTINE TUJOH (B11A124) SBS2 NUR AMALINA BINTI SHAHABUDIN (B11A294) SBS2 NURHAIZAN BINTI KAMARUZZAMAN (B11A323) SBS2 SITI SUBAILAH BINTI HAJI SUOD (B11A388) SBS2 WAN SHAHIRAH NERINI BINTI ROSLI (B11A423) SBS2 MUHSIN BIN ABD RAZAK (B11A251) SBT2 MOHAMAD IQBAL BIN ALI AKBAR (B11A233) SBS2 LAM CHIN JACK (B11A143)SBT2 LECTURER : DR FATIMAH KAYAT GROUP : 7 DATE OF PRESENTATION : 28TH MAY 2013
BANANA SUCKERS
INTRODUCTION OBJECTIVES 1)Providing us a basic understanding of physical and chemical requirements of cell, tissue, organ culture, their growth and development. 2)To learn and understand a procedure that is often used to propagate many plants of the same genetic background.
Banana belongs to Musaceae family, second largest food-fruit crops of the world produced in the tropical and subtropical regions. several constraints including highly season dependent and sometimes poor quality of planting material of banana become a limiting factor for banana propogation. Therefore, tissue culture technology was considered an appropriate option to overcome this problem. Application of tissue culture technology to propagate banana plants gives some advantages resulting in disease free planting materials, more vigorous growth, high yields, better quality of fruits, earlier fruiting and more uniform crop since they are made from selected high yielding mother plants.
The suckers are the major source of planting material for banana and normally remain true-totype. -sword suckers -water suckers
MEDIA PREPARATION MS MEDIA
200 ml of distilled water is added to a 500 ml conical flask 15g of sugar 25ml of Macronutrients stock 2.5ml of Micronutrients stock 1ml of Vitamin stock 2.5 ml of Plant Regulator Growth stock (Benzylaminopurine 5mg/L) 4g of agar is added Distilled water is added to adjusted the volume to 500ml.
Natural regeneration of cultivated bananas through suckers is very slow due to hormone-mediated apical dominance of the mother plant. So, as an alternative, shoot tips from young banana suckers with growing buds or cut rhizomes called bits and peepers are most commonly used as explants for accelerating the rate of in vitro propogation of banana.
MS MEDIA with 5mg BAP
IN-VITRO PROPAGATION (BANANA SUCKERS)
The suckers of banana tree is trimmed by discarding the outer layers of the pseudostem and roots leaving only shoot tips containing rhizome tissue 70% bleach and Tween-20 for (15 mins) spray the surface of laminar workflow with ethanol
The half trimmed banana suckers then sterile immediately by put in bottle contain 70% ethanol and shake it for 20 seconds and then 3 times of sterile distilled water.
Trimmed again
TISSUE CULTURE ON BANANA SUCKERS AND RICE CALLUS (MICROPROPAGATION)
RESULTS AND DISCUSSION
CONTAMINATED!! bacteria fungi (mycelium)
WHY TISSUE CULTURE Multiplying plant in short time and in large quantity Disease free plants Save space for early plantation/germination thus reduce cost for land used Cultured specific/usefull part of plant
PROCESS Sucker selection (sword sucker) Sterilization technique Growth media (MS media) Culturing Sub-culture
FACTOR Germination suitable nutrient composition (media) duration of alcohol use in sterilization Contamination Crowded work station Media/explant sterilization
IMPROVEMENT Container Sealing/ cap Media Working area/ method
SUGGESTION use shoot tips from young banana suckers with growing buds or cut rhizomes called bits and peepers are accelerating the rate of in vitro propogation of banana. Place the banana culture in complete darkness first for 1 week. add antioxidant in media such as ascorbis acid.
Callus Induction of Rice (Oryza sativa) Seeds
INTRODUCTION
Importance of Tissue Culture in Rice production Rice is the most important crop in Malaysia because it is staple food for Malaysian In Malaysia, rice demand is higher than production rate New technology must be developed to increase the rice production Tissue culture technology is one of the way to produce high quality rice which is can produce much rice grains, resistant to environmental stress and shorter development stage ( cut time so that can harvest quickly)
* Callus can undergo mutation to achieve the desired traits * The following methods are used to induce mutation in callus : 1. 2. Physical mutagens (irradiation) Chemical mutagens (carcinogens)
* The application of somaclonal variation in rice callus includes: 1. 2. For crop improvement 3. 4. To increase yield Producing plants that resistant to environmental stress To increase plant fertility
Description : Contaminated Factor : Technique of handling
Description : Contaminated Factor : Contamination from media
Formation of Callus - Compact callus
Callus Unorganized, undifferentiated, growing and dividing mass of cells. Form from the wounding part. Rapid plant multiplication for quality seed material. Performed in the dark as light can encourage differentiation of thecallus Important in plant biotechnology. Manipulationof the auxin to cytokinin ratio in the medium can lead to the developmentof shoots, roots or somatic embryos from which whole plants can subsequentlybe produced.
Callus Suspension culture Place friable callus into a liquid medium and agitate Single or small clumps of cells will be released into the medium, where they keep growing and dividing to eventually a cell suspension. The size of inoculum have to be sufficient , suspension can be build up faster. Too much cells and there is a risk that toxic products from damaged cells could accumulate to a inhibiting level. Cell suspension cultures should also be subcultured periodically.
CONCLUSION It can be concluded dichlorophenoxyacetic acid 2, 4- D is suitable for callus induction of rice seed. The role of 2,4-D in cell division is to increase the rate of cell division and this attributes to the increased amount of callus.
SUGGESTION REPEATED EXPERIMENTS. For callus induction MS medium supplemented with different concentrations of 2,4-D DIFFERENT VARIETIES OF RICE. ASD 16, ADT 43, Basmati 370, Pusa Basmati and Pokkali
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TISSUE CULTURE ON BANANA SUCKERS AND RICE CALLUS (MICROPROPAGATION)

  • 1. PLANT TISSUE CULTURE MEMBERS : KAREN LENGGIAU ANK AUGUSTINE TUJOH (B11A124) SBS2 NUR AMALINA BINTI SHAHABUDIN (B11A294) SBS2 NURHAIZAN BINTI KAMARUZZAMAN (B11A323) SBS2 SITI SUBAILAH BINTI HAJI SUOD (B11A388) SBS2 WAN SHAHIRAH NERINI BINTI ROSLI (B11A423) SBS2 MUHSIN BIN ABD RAZAK (B11A251) SBT2 MOHAMAD IQBAL BIN ALI AKBAR (B11A233) SBS2 LAM CHIN JACK (B11A143)SBT2 LECTURER : DR FATIMAH KAYAT GROUP : 7 DATE OF PRESENTATION : 28TH MAY 2013
  • 3. INTRODUCTION OBJECTIVES 1)Providing us a basic understanding of physical and chemical requirements of cell, tissue, organ culture, their growth and development. 2)To learn and understand a procedure that is often used to propagate many plants of the same genetic background.
  • 4. Banana belongs to Musaceae family, second largest food-fruit crops of the world produced in the tropical and subtropical regions. several constraints including highly season dependent and sometimes poor quality of planting material of banana become a limiting factor for banana propogation. Therefore, tissue culture technology was considered an appropriate option to overcome this problem. Application of tissue culture technology to propagate banana plants gives some advantages resulting in disease free planting materials, more vigorous growth, high yields, better quality of fruits, earlier fruiting and more uniform crop since they are made from selected high yielding mother plants.
  • 5. The suckers are the major source of planting material for banana and normally remain true-totype. -sword suckers -water suckers
  • 7. 200 ml of distilled water is added to a 500 ml conical flask 15g of sugar 25ml of Macronutrients stock 2.5ml of Micronutrients stock 1ml of Vitamin stock 2.5 ml of Plant Regulator Growth stock (Benzylaminopurine 5mg/L) 4g of agar is added Distilled water is added to adjusted the volume to 500ml.
  • 8. Natural regeneration of cultivated bananas through suckers is very slow due to hormone-mediated apical dominance of the mother plant. So, as an alternative, shoot tips from young banana suckers with growing buds or cut rhizomes called bits and peepers are most commonly used as explants for accelerating the rate of in vitro propogation of banana.
  • 9. MS MEDIA with 5mg BAP
  • 11. The suckers of banana tree is trimmed by discarding the outer layers of the pseudostem and roots leaving only shoot tips containing rhizome tissue 70% bleach and Tween-20 for (15 mins) spray the surface of laminar workflow with ethanol
  • 12. The half trimmed banana suckers then sterile immediately by put in bottle contain 70% ethanol and shake it for 20 seconds and then 3 times of sterile distilled water.
  • 17. WHY TISSUE CULTURE Multiplying plant in short time and in large quantity Disease free plants Save space for early plantation/germination thus reduce cost for land used Cultured specific/usefull part of plant
  • 18. PROCESS Sucker selection (sword sucker) Sterilization technique Growth media (MS media) Culturing Sub-culture
  • 19. FACTOR Germination suitable nutrient composition (media) duration of alcohol use in sterilization Contamination Crowded work station Media/explant sterilization
  • 21. SUGGESTION use shoot tips from young banana suckers with growing buds or cut rhizomes called bits and peepers are accelerating the rate of in vitro propogation of banana. Place the banana culture in complete darkness first for 1 week. add antioxidant in media such as ascorbis acid.
  • 22. Callus Induction of Rice (Oryza sativa) Seeds
  • 24. Importance of Tissue Culture in Rice production Rice is the most important crop in Malaysia because it is staple food for Malaysian In Malaysia, rice demand is higher than production rate New technology must be developed to increase the rice production Tissue culture technology is one of the way to produce high quality rice which is can produce much rice grains, resistant to environmental stress and shorter development stage ( cut time so that can harvest quickly)
  • 25. * Callus can undergo mutation to achieve the desired traits * The following methods are used to induce mutation in callus : 1. 2. Physical mutagens (irradiation) Chemical mutagens (carcinogens)
  • 26. * The application of somaclonal variation in rice callus includes: 1. 2. For crop improvement 3. 4. To increase yield Producing plants that resistant to environmental stress To increase plant fertility
  • 27. Description : Contaminated Factor : Technique of handling
  • 28. Description : Contaminated Factor : Contamination from media
  • 29. Formation of Callus - Compact callus
  • 30. Callus Unorganized, undifferentiated, growing and dividing mass of cells. Form from the wounding part. Rapid plant multiplication for quality seed material. Performed in the dark as light can encourage differentiation of thecallus Important in plant biotechnology. Manipulationof the auxin to cytokinin ratio in the medium can lead to the developmentof shoots, roots or somatic embryos from which whole plants can subsequentlybe produced.
  • 31. Callus Suspension culture Place friable callus into a liquid medium and agitate Single or small clumps of cells will be released into the medium, where they keep growing and dividing to eventually a cell suspension. The size of inoculum have to be sufficient , suspension can be build up faster. Too much cells and there is a risk that toxic products from damaged cells could accumulate to a inhibiting level. Cell suspension cultures should also be subcultured periodically.
  • 32. CONCLUSION It can be concluded dichlorophenoxyacetic acid 2, 4- D is suitable for callus induction of rice seed. The role of 2,4-D in cell division is to increase the rate of cell division and this attributes to the increased amount of callus.
  • 33. SUGGESTION REPEATED EXPERIMENTS. For callus induction MS medium supplemented with different concentrations of 2,4-D DIFFERENT VARIETIES OF RICE. ASD 16, ADT 43, Basmati 370, Pusa Basmati and Pokkali