Tissue culture rescue of a wild

Asteaceae disapeared in nature

Manuelle Bodin, Catherine Gautier, Stéphane Buord

IAPB 2014 – Melbourne

Cylindrocline lorencei Wild Asteraceae from Mauritius

First observed in XIXth in 2 distinct areas

Described in 1982

Considered as extinct since 1990

Key dates

Last seeds collected by JY Lesouef, botanist in

Conservatoire Botanique National de Brest, in

1982

End of the 80s, extinction in Plaine Champagne

Failure of cultivation of the plants in all Botanic

gardens in 1987

The 1rst rescue

In 1990, only seeds still conserved in

Brest, at -18°C

Embryo rescue for 74 seeds unable to

germinate

Only 6 plantlets obtained, 3 survived at

acclimatisation

2001, 3 clones in 2 Botanic Gardens in

the world

2003 : flowering but no viable seeds

Failure of cuttings

Cell biology lab in Vegenov

R&D lab,

Suppliyng services for breeders:

haplomethods,

embryo rescue,

tissue culture tools (shoot tip, protoplast, somatic embryogenesis)

Attempt of 2nd rescue by tissue

culture, with two constrains : Use the less plant material as possible

Avoid genetic variation in tissue culture

Starting from old stems …

Collect sprouting buds on the stems

Starting from old stems …

Collect sprouting buds on the stems

Careful desinfection

85 explants plated => 12 survived without contamination

MS salts + Morel vitamines

BPA 0,5 + NAA 0,05 mg/l

Sucrose 20 g/l

AC 1,5 g/l

A slow propagating process

No elongation of vitroplant stems

MS salts + Morel vitamines

2iP 1,5 mg/l

Sucrose 30 g/l

+/- AC 1,5 g/l

21°C, 16h light/day

1 month

darkness

A slow propagating process

No elongation of vitroplant stems

MS salts + Morel vitamines

2iP 1,5 mg/l

Sucrose 30 g/l

+/- AC 1,5 g/l

21°C, 16h light/day

> One year

Getting out of glass …

Rooting and acclimatisation

½ MS salts +

Morel vitamines

AIA 1 mg/l

Sucrose 20 g/l

21°C, 16h light/day

No soil !

On the way back to nature

The local partner:

2011 : One of the 30 endangered species sent back to

Mauritius

On the way back to nature

Preparing the last known habitat

On the way back to nature

Preparing the last known habitat

=> successful use of biotech for biodiversity

preservation purposes

Questions still to answer :

Will the plants survive in nature?

Will they be able to colonize this place?

Was Plaine Champagne the best habitat for this species?

In Vegenov In CBNB

New project for CBNB in Haïti

In relation with OPDFM

Juniperus ekmanii

Access to suitable

explants for tissue culture

Work supported by

Catherine Gautier

Bruno Bordenave

Stéphane Buord

Fanch Le Hir

Jean-Yves Lesouef

Nolwenn Quéniat

Estelle Cueff

Sonia Letty

Emilie Argouarch

Mikaele Paugam

Mickael Euzen

Murielle Philippot

Manuelle Bodin

Thank you for your attention

Une partie de ces travaux est

cofinancée par l'Union

européenne. L'Europe s'engage

en Bretagne avec le Fonds

Européen de Développement.