This document summarizes efforts to rescue the endangered plant species Cylindrocline lorencei from Mauritius through tissue culture techniques. The species was last observed in the wild in the 1980s and was considered extinct by 1990. In 1990, embryo rescue was used on the last remaining seeds, yielding only 6 plantlets. By the early 2000s, these 3 surviving clones were the only individuals left in botanic gardens. A second rescue effort in the 2010s used tissue culture on sprouting buds from old stems, producing 12 explants that survived without contamination. After over a year of slow propagation, rooted plants were produced. In 2011, one of the rescued plants was returned to its native habitat in Mauritius in

1. Tissue culture rescue of a wild Asteaceae disapeared in nature Manuelle Bodin, Catherine Gautier, Stéphane Buord
IAPB 2014 – Melbourne

2. Cylindrocline lorencei Wild Asteraceae from Mauritius First observed in XIXth in 2 distinct areas Described in 1982 Considered as extinct since 1990

3. 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

4. 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

5. 2003 : flowering but no viable seeds
Failure of cuttings

6. Cell biology lab in Vegenov
R&D lab, Suppliyng services for breeders: haplomethods, embryo rescue, tissue culture tools (shoot tip, protoplast, somatic embryogenesis)

7. Attempt of 2nd rescue by tissue culture, with two constrains :
Use the less plant material as possible
Avoid genetic variation in tissue culture

8. Starting from old stems …
Collect sprouting buds on the stems

9. 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

10. 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

11. 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

12. 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

13. No soil !
On the way back to nature
The local partner:
2011 : One of the 30 endangered species sent back to Mauritius

14. On the way back to nature
Preparing the last known habitat

15. On the way back to nature
Preparing the last known habitat

16. => 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

17. New project for CBNB in Haïti
In relation with OPDFM
Juniperus ekmanii
Access to suitable explants for tissue culture

18. 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.