In vitro culture for production of quality banana and plantain planting material
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A member of CGIAR consortium www.iita. org In vitro culture for production of quality banana and plantain planting material GUEYE Badara RTB in vitro propagation and conservation specialist (In vitro Genebank) International Institute of Tropical Agriculture (IITA) Headquarters: PMB 5320, Oyo Road, Ibadan, Nigeria Web: http://www.iita.org/genetic-resources-center 2 nd International Workshop of the Learning Alliance on Banana Bunchy Top Disease Control in Africa. 9 to 14 March 2015. IITA Ibadan, Oyo Road, Ibadan, Nigeria
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Transcript of In vitro culture for production of quality banana and plantain planting material
A member of CGIAR consortium
www.iita.org
In vitro culture for production of quality banana and
plantain planting material
GUEYE Badara RTB in vitro propagation and conservation specialist (In vitro Genebank)
International Institute of Tropical Agriculture (IITA)Headquarters: PMB 5320, Oyo Road, Ibadan, Nigeria
Web: http://www.iita.org/genetic-resources-center
2nd International Workshop of the Learning Alliance on Banana Bunchy Top Disease Control in Africa.
9 to 14 March 2015. IITA Ibadan, Oyo Road, Ibadan, Nigeria
A member of CGIAR consortium
www.iita.org
PTC gives opportunity to grow plant material in artificial conditions with advantages such as:
-Possibility for sanitation (culture in sterile conditions, pathogen cleaning)
-Rescue and conservation (continuous availability, space saving, cost-effectiveness, cryo)
-Micro-grafting and micro-propagation (mass multiplication: Bioreactor, cell suspension, polyshoot…)
-Germplasm exchange
-Door opened for more research (genetic transformation, somatic embryogenesis, haploid production, embryo rescue, molecular and pathological/virological investigations…)
INTRODUCTION
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Musa spp. GERMPLASM SANITATION
Vegetatively propagated crop: accumulation of pathogens over the reproductive cycles
3 major viruses: BBTV, BSV and CMV
Disease control: Production of clean planting material through virus eradication
Cleaning techniques through in vitro culture: Meristem regeneration, Thermo-treatment and Chemo-treatment
Factors: (a) the virus characteristics, (b) the type of tissues treated (c) the plant species(d) the genotype
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In vitro VIRUS ELIMINATION TECHNIQUES FOR Musa spp.
In vitro Virus elimination techniques
Meristem regeneration Thermo-treatment Chemo-treatment
(+)
- Most efficient on phloem-associated viruses ( BBTV)
- Association with other techniques
- « the smaller, the better »
- Mainly heat-treatment, cryo-treatment less popular
- Widely used in vivo/in vitro and associated with meristem culture
- A wide range of potentially antiviral molecules tested
- More efficient on CMV
(-) - Less efficient for virus in meristematic dome (CMV)
- Depends on the virus type
- mode of action not very known
- Time consuming and harmful
- Phytotoxicity and negative effect on regeneration
- Mode of action (inhibition, elimination, « silencing ») ???
Schematic representation of virus eradication processes for banana. The green , blue, and red arrows represent the process of chemotherapy, meristem culture, and
thermotherapy, respectively (Lassois et al., 2012. In Maurizio Lambardi et al. (eds.), In press)
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MICROPROPAGATION
Musa spp. are very responsive to in vitro PTC
Meristem regrowth in 4 to 6 weeks, followed by shoot development
Proliferation: subculture of fully developed plantlet
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MICROPROPAGATION
Musa spp. multiplication using semi-solid culture medium
Plantain/banana proliferation medium
Average multiplication rate onto semi-solid culture medium
4:1
Components Qty / LMS basal medium 4.43 g Myo-inositol 100 mg Sugar 30 g IAA (Indol-Acetic Acid) 0.18 mg BAP (Benzyl Amino Purine)
4.5 mg
Ascorbic acid 10 mgAgar 7 g
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Large scale multiplication
MICROPROPAGATION
DeRoi, SA
National genebank, SA
Vitropic, France
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MICROPROPAGATION
Musa spp. multiplication using Temporary Immersion System (TIS) Bioreactor
Rocket kit shaker & shouthern sun bioreactor
Air-lift Method
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MICROPROPAGATION
Musa spp. multiplication using Temporary Immersion System (TIS) Bioreactor
NACGRAB, Nigeria
Setis Method
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MICROPROPAGATION
Musa spp. multiplication using Temporary Immersion System (TIS) Bioreactor
Average multiplication rate using TIS
16:1
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HARDERNING
Acclimatization processMore than 95 % success
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HARDERNING
Nursery, a vital step (DeRoi, SA)
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Virus cleaning by meristem culture+ Most popular and more efficient (on phloem-associated viruses like BBTV)
+ « the smaller, the better »
+ Association with other techniques
e.g.: On Musa acuminata (AAA, cv Williams)
- BBTV eliminated at 99 % elimination
- BSV at 76 % and 41 % for in vivo and in vitro respectively
-Balance with regeneration capacity: « the bigger the better »
- Less efficient on CMV, virus in meristematic dome (1 à 7 % clean)
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Virus cleaning by Thermo-treatment
+ Mainly heat-treatment, cryo-treatment less popular
+ Widely used in vivo/vitro and associated with meristem culture
- Depends a lot on the virus type
- mode of action not very known
-Time consuming and harmful
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Virus cleaning by Chemo-treatment
+ A wide range of potentially antiviral molecules tested
+ More efficient on CMV
+ Example: Virazole ® (Ribavirin, 1-B-D-ribofuranosyl-1-2-4-triazole-3 carboxamide),
synthetic analogue of guanosine, replication inhibitor)
-Phytotoxicity and negative effect on regeneration
- Mode of action (inhibition, elimination, « silencing »)
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Recommandation: combination of techniques
- Meristem culture + thermo-treatment:
e.g.: On Musa acuminata (AAA, cv Williams)
- CMV eliminated at 38-70 % elimination
- BSV at 79 % and 67 % for in vivo and in vitro respectively
Banana and plantain virus cleaning
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Banana and plantain virus cleaning
CHALLENGES
- Clean or not expressed (« opportunism » of the virus)???
-What about B genomes???
- Integration of the virus in the genome (BSV)
- Inhibition of the expression (BSV)
- Policy issues (germplasm exchanges)…
Recommandation: combination of techniques
- Meristem culture + thermo-treatment:
e.g.: On Musa acuminata (AAA, cv Williams)
- CMV eliminated at 38-70 % elimination
- BSV at 79 % and 67 % for in vivo and in vitro respectively
Banana and plantain virus cleaning
Banana and plantain virus cleaning
CHALLENGES
- Clean or not expressed (« opportunism » of the virus)???
-What about B genomes???
- Integration of the virus in the genome (BSV)
- Inhibition of the expression (BSV)
- Policy issues (germplasm exchanges)…
