in vitro propagation of breadfruit (artocarpus altilis)

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IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

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Page 1: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

IN VITRO PROPAGATION OF BREADFRUIT

(Artocarpus altilis)

Page 2: 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”

Page 3: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

• 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.

Page 4: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

BENEFITS OF IN VITRO PROPAGATION

• 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

Page 5: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

• 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

CONSTRAINTS

Page 6: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

GENERALISED METHODOLOGYThe 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

Page 7: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

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)

TISSUE CULTURE MEDIA

Page 8: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

INITIATION OF STERILE CULTURES• In vitro cultures were initiated from shoot buds collected from Réduit

• The buds were thoroughly cleaned and surface sterilised

Page 9: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

• Cultures were then inoculated on Establishment Media

Page 10: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

• 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

RESULTS and OBSERVATIONS

Page 11: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

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 Media2. Too Severe Surface Sterilisation Procedures3. Release of Phenolics by explants

Page 12: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

MICROPROPAGATION• Mass production of Breadfruit plantlets was achieved

by transferring explants to Shoot Multiplication Media

Page 13: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

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

Page 14: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

Growth Room Conditions• Cultures were kept in Growth Room at a temperature of 25oC and a 16hrs photoperiod

Page 15: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

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.

Page 16: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

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

Page 17: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

• 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

Page 18: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

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

Page 19: IN VITRO PROPAGATION OF BREADFRUIT (Artocarpus altilis)

THANK YOU FOR YOUR ATTENTION