Somatic embryogenesis : control of somatic embryo maturation in hybrid larch

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Somatic embryogenesis : control of somatic embryo maturation in hybrid larch

Marie-Anne Lelu-Walter, Arnaud Lointier, Marie-Laure Gouez, Christophe Narboux, Daniel Cornu, Claude Joseph, Philippe Label

To cite this version:

Marie-Anne Lelu-Walter, Arnaud Lointier, Marie-Laure Gouez, Christophe Narboux, Daniel Cornu, et al.. Somatic embryogenesis : control of somatic embryo maturation in hybrid larch. 1. Meeting of the Cost 822 Working group, Apr 1995, Prague, Czech Republic. �hal-02778030�

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Comment citer ce document :

Lelu-Walter, M.-A., Lointier, A., Gouez, M.L., Narboux, C., Cornu, D., Joseph, C., Label, P.

(1995). Somatic embryogenesis : control of somatic embryo maturation in hybrid larch. In:

Mechanisms and markers of regeneration and genetic stability. Extended abstracts (p. 12-13).

Presented at 1. Meeting of the Cost 822 Working group, Prague, CZE (1995-04-27 - 1995-04-30).

EXTENDED ABSTRACTS

FIRST MEETING OF THE COST 822 WORKING GROUP

MECHANISMS AND MARKERS OF REGENERATION

AND

GENETIC STABILITY

PRAGUE

27APRIL - 30 APRIL 1995

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Somatie embryogenesis hybrid lareh

control of somatie embryo maturation ln

M·A. Lelu. A. Lointier, M.L. Gouez, C. Narboux, D. Cornu, C. Joseph, p, Label.

LN.R.A., Station J'Amélioration des Arbres Forestiers, F-45160 Ardon. Franœ

Aim

Somatiecmhl)'ogcllcsisofhyhtid larch(Larixxieptoeuropaea)bas hccn sucœssflllly dcvdnpcd al INRA of Orléans (France) sillec 1990. The maturation îs an important stcp as the gcnnination and the plant1er recoverics arc dcpcnding on the qualîty of thl.': somalie cmhryos. For the conifcrs. as for the angiospcnns. a great improvcrncnt was obtaincd by the addition ofah~l:i.sic

acid (ABA) 10 the cultuw medium. This gave fisc 10 wdl~dcvdopcd somalie cmbryos with typical morphology (considcring th~ zygotic embryo), synchronous developmenl and no precocious germination (1). Changes in content in ABA during maturation of hyhrid larch somalie embryos were followed using an enzyme immunoassay (ELISA).

Materials and Methods

Somatie embryogenic lines of hybrid larch (uJrix kaempferi xL. riecirilla) were ohlained in 1990 from immature zygotic emhryos and since suoculturcd evelY two weeks on hasal MSG mediulll as pn;viously dcscrihed (2). The maturation consislCd in the obtention of cOIylcdonary somali..:

emhryos well-devclopcd. The maturation medium consisling in MSG medium supplemenled with 0.2 M sucrose. 1~M ISA and 60 ~M ASA (2). Artel' 3 weeks of culture; cotyledonary somatic emhryos wen: transferrcd to germination medium whicb is a growth regulalnr-frce MSG medium. After 5 wœks of germination, germinatcd somalie emhryos which devduped need1cs were considered as plantlcLS. For the hormonal analyses, the somatic emhryos wll1l.:h undcrwcnt cmhryogeny were removed al'ter 0, l, 2, 3, 4 and 5 wecks of maturation. Analyses using an enzyme immunoassay (ELISA) were realised as previously descrihcd (1).

Results

Developmcnt of cotylcdonary somalie embryos was greaùy improwd hy the pl"èSenCC ^01"ASA ¹¹¹ the culture mèdium (1). l-Iowever in hyhrid lareh. the extension of Ihe period ^01" malllrJ.lioll in presence of ASA from ::\ wecks to 4 or 5 weeks œsultcd in a significant dCl:rcase ln hoth germination and plantlct frequcncics (figure 1). The highest germination rates ^(9()Ç() \Vere obtained for 3 wccks-old somatie embryos. In contrast prolongation of maturation pcJiod 10 5 wccks rcsu1ted in œduced gCl1l1ination (38%); this was aiso obscrvcd for the plantlct rccovery : the development of the aerial pans was reduced from 70 to 10% for respectively 1and 5 wcàs- nid somatic cmbryos. In presence of ASA in the culture medium, the ASA levcl of the sornatil:

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emhryos increased during their dcvelopmcnt 10 reach a maximum valllc at \Vcck 5 (15.~

pmoUcmbryo. figure 1). As a consequence. wc assume that il was the levcl of ABA in p/mllo Ihal \Vas fèsponsihk for the inhihition.

Conclusion

ABA actcd in both a stimulatory and an inhibitory manncr. If ABA promoted the ohtention nf bigh quality snmalic cmhryos in large numbcrs. il also had a dcktcrious effeel on the suhscqw.:nt dcvelopmcnt. i.e. genninalion and plantlcl recovcries. The rcsults stresscd lhe importance of the maturation procedure and duration.

Literature

(1) LE LU MA and LABEL P (1994). Physiol. PianI.. 92. 53-60.

(2) LELU MA. BASTIEN C. KLiMASZEWSKA K. WARD C and CHAREST P (1'!94). Plant Ccii Tissue Organ Culture. 36. 107-115.

1

200

50 \00 150

ABA (pmoUembryo)

oo

0,9

0,8

0,7 0,6 0,5 0,4

0,3

~ 0,2

~ ^0,1

~...

Figure 1: Rclalionship octwœn cndogenous ABA levcls and gro\Vth rates of somatÎl' cmhryos.

• : germination rate at 28 days of culture;. plantlet raie at 28 days of clIltllrc.Bar.l. rcprescnt standard errors on the mean (n=5 for ABA. n=60 for rates).