Transcript In Vitro - paepardmauritius

IN VITRO PROPAGATION OF
BREADFRUIT
(Artocarpus altilis)
INTRODUCTION
• In Vitro propagation (tissue culture) offers
prospects for rapid bulking up of Breadfruit
planting material
• Usually vegetatively propagated using shoot
or root cuttings and is essential for
multiplication of seedless varieties
• Seeds: are rarely planted because of genetic
variability and viability
• Research is being conducted at the FARC
Tissue Culture Section, Réduit on Breadfruit “In
Vitro Mass Propagation”
• Aims: Development of a commercially efficient
Micropropagation Protocol for Breadfruit
• Expected to contribute to Local Sustainable
Resources for Agriculture, Backyard Planting,
Agro-forestry, Reforestation and ultimately
Food Security
• Possibility for safe trans-boundary movement
towards other countries interested in
Breadfruit farming.
BENEFITS OF IN VITRO PROPAGATION
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FAST: More plants produced in a given period
Plantlets produced are genetically identical
Plants are produced under sterile conditions
Year round production of plantlets and not
dependent on external environment
• Little space requirement for multiplication
• Propagated material can be stored for a long
time
• Little attention required for In Vitro cultures
between subcultures
CONSTRAINTS
• Requirement for specialised production
facilities
• Propagation Protocol development is time
consuming
• Plantlets obtained are small and juvenile
• Genetic Variability can occur upon excessive
sub-culturing
• Transitional phase required for adaptation of
plantlets to In Vivo conditions
GENERALISED METHODOLOGY
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The General steps undertaken for the In Vitro
Mass Propagation protocol development
were:
Explant Collection and Surface Sterilisation
Establishment of Aseptic Cultures
Shoot multiplication
Rooting of shoots
Acclimatisation of Rooted TC Plantlets
TISSUE CULTURE MEDIA
Three Culture media have been Successfully
formulated at the FARC Tissue Culture Section
based on Literature Leads available
These media are referred to as:
1. Establishment Media (EM)
2. Shoot Multiplication Media (SM)
3. Rooting Media (RM)
INITIATION OF STERILE CULTURES
• In vitro cultures were initiated from shoot buds
collected from Réduit
• The buds were thoroughly cleaned and surface
sterilised
• Cultures were then inoculated on Establishment
Media
RESULTS and OBSERVATIONS
• Four trials were undertaken for development
of an efficient Surface Sterilisation Protocol
• Success rate of the Selected Sterilisation
Protocol was 85 % after modifications brought
• Contamination was mainly of bacterial origin
MORTALITY
• Despite successful initiation of Aseptic Cultures
death of explants was a major drawback during trials
• Mortality from trials due to explant necrosis were
principally because of:
1. Unsuitable Establishment Media
2. Too Severe Surface Sterilisation Procedures
3. Release of Phenolics by explants
MICROPROPAGATION
• Mass production of Breadfruit plantlets was achieved
by transferring explants to Shoot Multiplication
Media
SUBCULTURE
• Sub-culturing of plantlets was done every 5-7 weeks
• Axillary branches and buds were excised from main
shoots and inoculated on fresh media
• An overall multiplication rate of 2.5-3 was observed
Growth Room Conditions
• Cultures were kept in Growth Room at a
temperature of 25oC and a 16hrs photoperiod
ROOTING OF PLANTLETS
• Rooted plantlets were obtained 5-6 weeks after
inoculation on Rooting Media
• However, Root development on Rooting Media was
observed for only about 60% of plantlets.
HARDENING
• Rooted plantlets of about 5cm in height were
selected for hardening
• Trials have been performed on different
hardening media ( Soil, Rocksand and Perlite)
• Plantlets were kept in the ECU for adaptation
to In Vivo conditions
• Hardening is expected to last between 5-8 mths
• Leaf Margins were smooth in In Vitro cultures
compared to the typical Lobed leaf morphology
under In Vivo conditions
• Hardening could be considered successful as
soon as plantlets develop the typical lobed
leaves characteristic
FUTURE WORKS
• Fine tuning and Optimisation of protocol
• Reducing production time and cost of
plantlets production
• Assessment of genetic fidelity of plantlets
after successive sub-culturing
• Optimisation of conditioning & hardening for
higher % survival
• In Vitro conservation of Breadfruit Varieties
THANK YOU FOR YOUR
ATTENTION