lect_pence.pdf

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n or c en c ongress

Challenges in Botanical Research and Climate Change

Supplementing Seed Banks:

- -- ,

In Vitro Methods for Preserving and Propagating

Species at Risk

Valerie C. Pence

Center for Conservation and Research of Endangered Wildl ife (CREW)

nc nna oo o an ca ar enCincinnati, Ohio, U.S.A.


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Cincinnati, Ohio, USA

Population:

City: 365,000

Area: 1.9 million,

includin areas in

the states of Ohio,Kentucky, and

Indiana


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Cincinnati Zoo & Botanical Garden

Cr e a t i n g A d v e n t u r e , Co n v e y i n g K n o w l e d g e , Co n s e r v i n g N a t u r e , Se r v i n g Com m u n i t y

Second oldest Zoo

in the U.S. (1875)

Now also aBotanical Garden

(1987)


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Research Center- CREW

Center for

CConservation and

EEndangered

WWildlife

Mission: To use science and technology to understand, preserve and propagate


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Plant Research Division

CREW

Endangered Plant

Goal: To use in vitro methods for propagating

and preserving endangered plants when

traditionalmethods are not adequate


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In vitro methodsfor propagating and preserving endangered plants

In Vitro Collectin


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Tissue culture propagation


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Cryopreservation

Frozen Garden-

EPPP Tissue Bank

Pteridophyte Bank

Bryophyte Bank


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Center for Plant Conservation -Collaborators

Over 40 targeted species 15 collaborating gardens, and others . . .


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Rising sea levels

Drought

Loss of habitat

Small populations vulnerable

to catastrophic events

Ex situ conservation can be used to back up and support wild populations


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-

Seed/Spore

Banks


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But. . .ee pore an ng

Efficient storage of

multiple genotypes in a

small space

Applicable to many Species with few or no

and most spores)seeds

Species with

recalcitrant seeds


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Embr o cr o reservation Shoot ti cr o reservation

Species with recalcitrant seeds

Species with no seeds or

recalcitrant seeds


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Using In Vitro Methods

Three examples . . .


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Hedeoma todsenii

Todsens pennyroyal

Lamiaceae

South central New Mexico, 6,000-7,000 ft

Collaborator: The Arboretum at Flagstaff

ounta n a tat ro uces tt e or no see


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Hedeoma todsenii

Tissue culture propagation protocols


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Hedeoma todsenii Cryopreservation and Banking

35

40

45

val

20

25

30

V

D

rce

ntsurv

0

5

10P

Shoot tips

banked

Average survival of shoot tips, two weeks after exposure to

li uid nitro en usin two methods: D = enca sulation

% SurvivalMedium (seeds notavailable)

dehydration (n=10); and V=encapsulation vitrification (n=24).


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Hedeoma todsenii - Genetic Analysis

Interfering

compounds

inhibit DNAana ys s, excep

with in vitro

material

A (left): Most parsimonious tree made from RAPD

. .

population sources for the genetic lines. Matching

colors in A and B identify the population source.


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Ranunculus aestivalis

Autumn buttercup

Ranunculaceae

ou en ra a

Collaborator: The Arboretum at

Flagstaff

One population declining to about

20 plants

Wet meadow habitat; h drolo

changing?


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Ranunculus aestivalis

Outplanting, June, 2007

Acclimation:

The Arboretum at Flagstaff

Propagation:

Outplanting:

TNC Sevier

CREW Valley Preserve

Utah Valley

State College

USFWS funding


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Ranunculus aestivalis - outplanting

138 plants augmenting approx 20

plants remaining in the wild

Half lanted with wild lants half in

wetter area

92% survival through the summer

Wetter site doin better


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Asimina tetramera -

Four-petal pawpaw

Annonaceae

Southeast Florida - scrub habitat

Collaborator: Historic Bok

Sanctuary

Endemic to two counties

Occurs only 15 - 35 ft above sea

level

Recalcitrant seeds


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Asimina tetramera -

Protocols

Propagation

Cryopreservation


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Asimina tetramera - Germplasm banking

In vitro collecting Shoot propagation

Cryostorage

analysis


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Conclusions

Cryopreservation of in vitro propagated

material can su lement seed bankin for s ecies

with little or no seed or with recalcitrant seeds

In vitro propagated plants can be used to test

,

In vitro propagated plants may allow genetic

analysis when wild tissues are not suitable


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Plant Division Staff(past and present)

Kristine LindseyPeter Ackerman

. . .

and Students

Dr. Doug Winget

Bernadette Plair


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Thank you for your attention!

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