.v. •.._...••• -.. •••..---- .. ----•.. ••• •••BIOTECHNOLOGIES•• Oil palm biotechnology: progress and prospects* Alain RIVAL " [arnes TREGEAR 2, Estelle JALlGOT 2, Fabienne MORCILLO 2, Frederique ABERLENC 2, Norbert BILLOTIE " Frederique RICHAUD 2, Thierry BEULE 2, Alain,'BORGEL 2, Yves DUVAL 2 I Cirad-CP. TA80/PSfll, bd de la Lironde, 34398 Montpellier Cedex OS, France <alain. [email protected]> 2 Cirad-CP/IRO, UMR 1098, Centre IRO, BP 5045, 34032 Montpellier Cedex 01, France Abstract: Today, a range ofbiotechnological approaches, from somatic Resume: Les strategies d'amelioration genetique dupalmier ahuile (Elaeis embryogenesis tobiomolecular research, play anincreasingly important role guineensis jacq.) font desormais appel atout un ensemble d'approches in breeding strategies for oil palm (Elaeis guineensis jacq.). biotechnologiques, allant deI'embryogenese somatique ala biologie mole­ Clonal micropropagation. Methods ofcloning byinvitro culture led tothe culaire. development ofa micropropagation technique for oil palm based onsoma­ Micropropagation clonale. Les methodes de cionage par culture in vitro ticembryogenesis which was tested at thepilot stage onelite genotypes, antconduit audeveloppement d'une technique de micropropagation qui a thus enabling theproduction ofhigh oil yielding clones. This phase allowed etetestee aI'echelle pilote sur des genotypes d'elite, permettant ainsi la the identification of limiting factors associated with scaling-up, with respect production declones hautement producteurs enhuile. Cette phase a revele in particular tothescale ofmass production required tomeet theneeds of uncertain nombre de facteurs limitants lies auchangement d'echelle, en planters and to theproblem ofensuring genetic fidelity in theregenerated particulier les productivites necessaites pour satisfaire la demande des plan­ plant material. These two concerns led researchers tolook further into the teurs et la fidelite genetique du matetiel vegetal regenere. Ces deux pro­ underlying physiological and/or molecular mechanisms involved in soma­ blemes antconduit Jes chercheurs aanalyser les mecaaisme: physiologiques tic embryogenesis andthesomaclonal variation events induced bythein et/ou moiecutaues impliques dons I'embryogenese somatique et les varia­ vitro cloning procedure. tions somaclonales induites parle clonage in vitro. Structural and functional genomics. Marker-assisted breeding in oil palm Cenomique structurale et fonctionnelle. La selection assistee par mar­ isa long-term multi-stage project including: molecular analysis of genetic queurs constitue, pour le palmier ahuile, unprojet along terme, articule diversity in both E. guineensis and E. oleifera germplasms; large scale deve­ sur plusieurs phases: I'analyse moleculaire de ladiversite genetique dons lopment of PCR-based microsatellite markers; and parallel development of le germplasm d' E. guineensis et d' E. oleifera ; le deveicppemen: agrande three genome mapping and QTL detection projects studying key agrono­ echelle par PCR de marqueurs microsatellites, et le developpement enparal­ mic characters. lele de trois projets de cartographie genetique etde detection de QTLs s'adres­ Post-genomics. In order to tackle theproblem ofthemantled flowering sant ades coroaeres agronomiques majeurs. abnormality, which isinduced during themicropropagation process, stu­ Post-genomique. Afin de comprendre le phenomene d'anomalie florale « mant­ dies ofgene expression have been carried outin tissue cultures as a means led ", induite aucours du procMe demicropropagation, des etudes sur l'ex­ ofestablishing anearly clonal conformity testing procedure. It isimportant pression du genome ant ete menees sur des cultures de tissus, dons lebut de toassess what kind ofmethodology is the most appropriate for clonal confor­ mettre aupoint untest de conformite preaxe. 11 importe d'evaluer quelle mity testing bycomparing RNA, protein and DNA (PCR) based approaches. methodologie estla plus adequate pour cetest, en comparant des approches Parallel studies ongenomic DNA methylation changes induced by tissue basees sur I'analyse des ARN, des proteines ou de I'AON (PCR). Les recherches culture suggest that thelatter may play animportant role in thedetermi­ menees en parallele sur les changements de niveaux de methylation de I'AON nation ofthemantled abnormality. genomique induits par culture in vitro suggerent une implication decisive de cephenomene dons le detetmimsme de I'anomalie « mantled ». Key words: Elaeis guineensis jacq., epigenetics, genomics, marker-assis­ Mats cles : Elaeis guineensis jacq., epigenetique, embryogenese somatique, tedbreeding, molecular markers, somatic embryogenesis. genomique, marqueurs maecutoues, selection assistee par marqueurs. * Results presented in the present article arepart of anoralcommunication givenat PIPOC 2001 (MPOB International Palm Oil Congress), 20-23August 2001, Kuala Lumpur, Malaysia. OCL VOL. 8 W 4 JUILLETJAOOT 2001 295 ..... Challenges in oïl seeds by c10ning the best male parents (pisi­ duction scheme currently used at Cirad/IRD palm biotechnology fera), since pollen production can be alimiting involves 4 distinct stages, as shown in Figure 1. factor [S, 6]; To date, embryogenic suspensions have been - the exploitation of (E. guineensis x E. o/eifera) successfully isolated for more than 20 different Due to the increasing demand for palm oil, the interspecific hybrids, a limited number of which c1onallines. The average concentration was lack of plantable land and the foreseeable are fertile, but which can show agood tolerance found to be ca 105 cell c1usters per litre with a increases in cultivation costs, it is necessary to to pests and diseases, notably in South America multiplication factor reaching 4x per month. make available to the planters planting mate­ [7]; These characteristics allow mass propagation. rial with high genetic potential. - the production of biclonal seeds from soma­ For oil palm, Sondahl (pers. com.) estimated the Nowadays, planting material consists solely of tic embryogenesis-derived parents [5]; price of an encapsulated embryo produced by tenera hybrids (fruits with shell of intermediate - the true-to-type regeneration of future gene­ bioreactor technology inthe USA at ca 0.20 USD, thickness), originating from crosses between tically-engineered materiel bearing useful agro­ to which has to be added the cost of in vitro cul­ dura (thick shell) and pisifera (thin shell) types, nomic traits [8]. ture for germination, rooting and acclimatisa­ the thickness character being controlled by a The biological characteristics of the oil palm do tion. Field trials are under way for the assess­ monofactorial gene [1]. not allow its vegetative propagation by conven­ ment of the clonai fidelity of plantlets originating The breeding strategies developed by seed com­ tional horticultural means, therefore, the only from cell suspension cultures. The first results panies are aimed at producing dura x pisifera possible way to c10nally propagate elite oil palms obtained from suspension culture-derived clones hybrids with a high productivity of oil contain­ is by means of somatic embryogenesis. C10ning are quite encouraging, and various research ing ahigh proportion of unsaturated fatty acids, of oil palm (Elaeis guineensis jacq.) is performed groups from the oil palm industry in Malaysia a low growth rate and, in certain cases, resis­ by inducing somatic embryogenesis on calli deri­ have now developed embryogenic suspension tance/tolerance to diseases such as vascular wilt ved from various tissue sources, using tissue cul­ cultures growing on 2,4-0 free or even auxin­ caused by Fusarium oxysporum elaeidis in Africa ture protocols recently reviewed in detail [8]. free media. These protocols are presently under [2]. Breeding schemes now incorporate the assessment and initial observations of trueness­ exploitation of genetic resources able to pro­ Current protocols to-type have revealed agenerally very low pero vide tolerance/resistance to diseases which are centage of abnormal palms. A management stra­ confined to specifie parts of the ail palm culti­ Approximately 25 years ago, two major groups tegy, based on the sequential cryopreservation vation area: Ganoderma disease in South East initiated research programmes for oil palm of embryogenic lines [18] cou Id be followed in Asia; and also bud rot in Latin America. micropropagation: in the UK and in Malaysia, order to lower the risk of losing embryogenic Since each selection cycle lasts for around the Unilever Plantations and Harrison and capacity in suspensions during multiplication. 10 years, genetic improvement is very slow, Crossfields Plantations group [9, 10] and in even if much progress has been achieved over France and Côte d'Ivoire, the IRHO/Orstom Embryo maturation the last 50 years [3]. Avery high heterogeneity group (which became Cirad-CP/IRD in the is still observed among hybrids, some palms 80's) [4, 11]. These programmes were initiat­ ail palm somatic embryos produced from producing 60% more oil th an the average of ed to complement in-house breeding strate­ embryogenic suspension cultures exhibit an the progeny of agiven cross [4]. These charac­ gies and were aimed at multiplying the elite incomplete maturation and develop directly teristics must be considered along with the low germplasm available in the plantations for com­ towards germination without passing through planting density (generally ca 143 palms per mercial use. In a recent review [8] a descrip­ aquiescent phase