Plant regeneration from embryogenic suspension cultures of oil palm (Elaeis guineensis Jacq.)

Suspension cultures of oil palm (Elaeis guineensis Jacq.) were established in a medium with 80 or 100 mg.l^–1 2,4-dichlorophenoxyacetic acid and 1 g.l^–1 activated charcoal, from calli producing embryogenic cells and protruding proembryos. The suspension was composed of meristematic clumps, breaking away and giving rise to new smaller aggregates. Under the best conditions, the initial weight increased about 4 fold in one month. Embryo differentiation was achieved when plating the clumps either directly, or after a phase in hormone-free liquid medium. In the second case, early maturation occurred in liquid medium. Secondary embryogenesis was reduced, and up to 18.1 % embryos formed shoots. Rooted plants were sucessfully transferred to soil.Abbreviations BAP 6-benzylaminopurine - NAA -naphthaleneacetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid     

Midday Closure of Stomata in the Oil Palm Elaeis guineensis. Jacq.

Under natural climatic conditions in West Africa, midday closureof stomata in 3–6-year-old oil palms occurs only in thelatter half of the dry season, when soil moisture is low andair temperatures are high. Using an infiltration method, attempts were made to determinethe climatic factors which affect stomatal closure. The extentof midday closure is related closely to air temperature andshade temperature under the palm, agreeing closely with thefindings of Meidner and Heath (1959) on onion, but the aperture-temperaturecurves are different for watered and unwatered palms in thedry season, the difference being greater at higher temperatures.Watering reduces closure by changing the aperture-temperaturerelationship. Under natural conditions this effect is usuallyobscured because of lower temperatures in the dry season followingrain. During Harmattan weather, with very low relative humidities,the aperture-temperature relationship broke down. The results are discussed in relation to current views on middayclosure of stomata.     

Morpho-agronomic and molecular characterisation of oil palm Elaeis guineensis Jacq. material from Angola

The characterisation of phytogenetic resources is used to improve conservation strategies, promote new sources of plant material, and design breeding strategies. In this study, we evaluated oil palm material with nine morpho-agronomic traits and 30 microsatellite markers (Simple Sequence Repeats; SSRs) that had been previously collected in five geographical regions of Angola. The analysis of variance for components of bunch production and oil yield showed highly significant (p?<?0.001) statistical differences between geographical regions and among families for all traits evaluated. The SSRs were highly informative, suggesting high genetic diversity (H _T ?=?0.666) among the accessions evaluated. However, the clustering pattern at both morpho-agronomic and molecular levels did not match the geographical distribution of accessions, showing a low genetic differentiation (G _ST ?=?0.039) between regions. On the other hand, genotypic (G _ST ?=?0.150) and phenotypic differences were found among families, which could offer the potential for future genetic gains in the oil palm. The information generated indicates that the evaluated accessions have desirable characteristics that should be included in breeding programs, which could expand the genetic basis of the crop.     

Occurrence of aromatic cytokinins in oil palm (Elaeis guineensis Jacq.)

The natural occurrence of 6-benzylaminopurine, 6-(2-hydroxybenzylamino)purine (ortho-topolin), 6-(3-hydroxybenzylamino)purine (meta-topolin), their ribosides and 9-glucosides is reported using specific antibodies to these groups of compounds in high performance liquid chromatography/enzyme-linked immunosorbent assay (HPLC/ELISA). Compounds were identified by their retention times and differential cross-reactivities with six antisera in analyses carried out in two laboratories using different HPLC gradient systems. Identities were confirmed by immunoaffinity purification followed by HPLC with on-line UV spectrum analysis. Further confirmation of the occurrence of ortho-topolin riboside and isopentenyladenine-9-glucoside was obtained from gas chromatography-mass spectrometry analysis of permethylated HPLC fractions of an extract of oil palm tissues. The aromatic cytokinins, and in particular, ortho-topolin riboside, were found in a variety of oil palm tissues at concentrations exceeding those of the isoprenoid cytokinins, zeatin, isopentenyladenine, dihydrozeatin, and their ribosides. The 9-glucosides of isopentenyladenine and zeatin were more abundant than those of the aromatic types. The cross-reactivity of benzyladenine compounds with antibodies to isopentenyladosine is discussed in relation to the interpretation of ELISA data.Abbreviations BA N ⁶-benzyladenine - HPLC high performance liquid chromatography - ELISA enzyme-linked immunosorbent assay - GC-MS gas-chromatography-mass spectrometry - mT meta-topolin - oT ortho-topolin - TEAA triethylammonium acetate - IAC immunoaffinity chromatography - SPD spectral photodiode     

Assaying lipase activity from oil palm fruit (Elaeis guineensis Jacq.) mesocarp.

The mesocarp of mature oil palm fruit undergoes intensive triglycerides hydrolysis upon abscission and bruising. This generates such a high amount of free fatty acids that the oil might become unfit for human consumption without appropriate refining. The lipase (EC 3.1.1.3) involved in the breakdown of the oil is not stable after homogenization of the tissue in aqueous buffers. In this study, we have devised a solvent-based procedure that allowed us to obtain fractions with stable lipase activity. Using these fractions, we have determined the optimal conditions for assaying mesocarp lipase activity. The activity was highest at a temperature of 35 degrees C and a pH of 9. The lipase was found to be strictly calcium dependent. The specific activity of the lipase measured in optimal conditions was found to be 33 mumol fatty acids released min(-1) mg(-1) protein using olive oil as substrate. The mesocarp contains about 190 U of lipase g(-1) fresh weight. This activity was found to be inhibited by the lipase inhibitor tetrahydrolipstatin (THL), suggesting that the lipase is a serine hydrolase.     

Quantitative trait loci for yield components in oil palm (Elaeis guineensis Jacq.)

The development of an oil palm RFLP marker map has enabled marker-based QTL mapping studies to be undertaken. Information from 153 RFLP markers was used in combination with phenotypic data from an F₂ population to estimate the position and effects of quantitative trait loci (QTLs) for traits including yield of fruit and its components and measures of vegetative growth. The mapping population consisted of 84 palms segregating for the major gene influencing shell thickness. Marker data were analysed to produce a linkage map consisting of 22 linkage groups. The QTL mapping analysis was carried out by interval mapping and single-marker analysis for the unlinked markers; significance thresholds were generated by permutation. Using both single-marker and interval-mapping analysis significant marker associated QTL effects were found for 11 of the 13 traits analysed. The results of interval-mapping analysis of fruit weight, petiole cross section and rachis length, and ratios of shell:fruit, mesocarp:fruit and kernel:fruit indicated significant (P<0.05) QTLs at the genome-wide threshold. The putative QTLs were associated with between 8.2% and 44.0% of the phenotypic variation, with an average of 27% for the single-marker analysis and 19% for the interval-mapping analysis. The higher percentage of phenotypic variation explained in the single-marker analysis, when compared to the interval-mapping analysis, is likely to be due to the lower stringency associated with the single-marker analysis. Large dominance deviations were associated with a sizeable proportion of the putative QTLs. The ultimate objective of mapping QTLs in commercial populations is to utilise novel breeding strategies such as marker-assisted selection (MAS). The potential impact of MAS in oil palm breeding programmes is discussed. Received: 26 June 2000 / Accepted: 24 October 2000     

Identification of p.Methoxyallybenzene in the Pollen of the Oil Palm Elaeis guineensis Jacq.

The lipid extract prepared from the pollen of the West African,oil palm, Elaeis guineensis, was found to contain p-methoxyallylbenzenein substantial amount. The pure substance after isolation bymeans of preparative gas-liquid chromatography and steam distillationwas characterized by using a combination of g.l.c.-mass spectrometry,infrared spectrometry, and chemical reactions. Mass spectrometrygave a spectrum with a parent ion at m/e 148 corresponding tothe molecular weight and fragmentation ions at m/e 133, 121,117, 105, 91, and 77. The structure was further confirmed bymeans of the infrared spectrum which showed major peaks at 1250,920, and 815 cm^–1 corresponding to a CH₃O group, a 1,4-substitutedaromatic ring, and a RCH=CH₂ group respectively. Its chemicaland physical properties (hydrogenation, oxidation, etc.) werefound to be similar to those of its positional isomer, p-methoxy--phenylpropene.The physiological and economic importance of the compound arediscussed.     

Cell Separation and Anatomy of Abscission in the Oil Palm, Elaeis guineensis Jacq.

Shedding of the fruit of the oil palm takes place in two co-ordinatedstages. The first, a cell separation event at a pre-defined,positionally differentiated abscission zone at the base of thefruit, is followed by further cell separation in peripheraltissue at the junction with the rudimentary androecial ringand the tepals. The position of the second separation is determinedby the age and ripeness of the fruit and the degree of pressureto which it is subjected; it is also dependent upon completionof the first stage. Implications of this unusual two stage separationprocess are discussed. Key words: Abscission, cell separation, anatomy, oil palm, Elaeis guineensis     

Viability and Storage of Pollen of the Oil Palm, Elaeis guineensis Jacq

The effects of drying pollen of the oil palm, Elaeis guineensis,and storage under various conditions, on its viability havebeen examined. Viability was based on the ability of pollento germinate on a sucrose medium with and without added boricacid, and on its capacity to set fruit when applied to receptivefemale flowers. Oven-drying at 37 °C for 2–8 h followedby storage in a deep freezer proved to be the best method ofstoring the pollen. Pollen treated in this way and stored for12 months was capable of producing fruits which were equal inweight and appearance to those produced by fresh pollen. Elaeis guineensis, oil palm, pollen, storage, viability     

Morphogenetic Potential of Embryo- and Seedling-Derived Callus of Elaeis guineensis Jacq. var. pisifera Becc.

The oil palm of commerce, Elaeis guineensis (tenera), derivesfrom hybridization of variety dura x pisifera. Germ plasm ofpisifera is limited, however, because of varying degrees offemale sterility. Efforts to increase the range of availablepisifera male germ plasm have thus far been limited to fertilepisiferas but it is anticipated that clonal multiplication ofembryos and explanted organs from less female-fertile, but nonethelessfertile, male, plants could play a significant role in breedingprogrammes. Callus has been produced from mature pisifera embryosand leaf explants from young plantlets using a half-strengthMurashige and Skoog medium supplemented with inositol, caseinhydrolysate, activated charcoal and varying concentrations ofauxins. After sub-culture on media of similar composition, organizedstructures can develop. Those which form in stale nutrient media(e.g. left in the same medium for up to 3 months) give riseto embryonal structures which do not readily develop furtherand can be regarded as neomorphs. If callus once formed is transferredto media with lower auxin concentrations (40–0 mg l^–1)embryonal buds and well-formed shoots are produced; additionof gibberellic acid (GA₃) fosters the production of well-organizedshoots accompanied by roots. Elaeis guineensis var pisifera, oil palm, tissue culture, micropropagation, embryo development, morphogenetic potential, callus     

Architecture and development of the oil-palm (Elaeis guineensis Jacq.) root system

The growth dynamics and architecture of the oil-palm root system are described. Following a transitional juvenile phase, eight different morphological types of roots have been distinguished according to their development pattern and state of differentiation: primary vertical and horizontal roots, secondary horizontal roots, upward growing secondary vertical roots and downward growing secondary vertical roots, superficial and deep tertiary roots and quaternary roots. The relative position of these types of roots determines a morphological and functional unit of the root system called 'root architectural unit' of the oil palm. This root polymorphism enabled us to define a morphogenetic gradient, which reflected the oil-palm root-system ontogenesis.     

Construction of a RFLP genetic linkage map for oil palm (Elaeis guineensis Jacq.).

We have developed a restriction fragment length polymorphism (RFLP) genetic map in oil palm (Elaeis guineensis Jacq.) for use in breeding programmes. A segregating population of 98 individuals was probed with 84 informative low copy clones (mainly PstI genomics). This yielded 103 scorable loci, of which 97 could be linked into 24 groups of two or more markers (n = 16 for oil palm), encompassing a total of 860 cM. The high level of linkage between markers (95%) suggests good genome coverage and very little segregation distortion of markers was observed. The mapping population, which was generated by the selfing of an important breeding palm (A 137/30), also segregated for the shell thickness character (Sh), enabling mapping of the gene conferring this major commercial trait. The nearest RFLP marker (pOPgSP1282) was located 9.8 cM from Sh in the mapping population and 6.6 cM away in a related (A137/30 x E80/29) smaller population (45 palms). Strategies for conversion of RFLP markers to a PCR format were explored and two important markers were shown to generate allelic variants following PCR amplification and restriction digestion of PCR products. The application of this map to oil palm breeding programmes is discussed.     

Somaclonal variation in oil palm (Elaeis guineensis Jacq.): the DNA methylation hypothesis

Elaeis guineensis Jacq.) currently hampers the scaling-up of clonal plant production. In order to investigate the relationship between the “mantled” somaclonal variant and possible alterations in genomic DNA methylation rate, two complementary approaches have been used. HPLC quantification of relative amounts of 5-methyl-deoxycytidine has shown that global methylation in leaf DNA of abnormal regenerants is 0.5–2.5% lower than in their normal counterparts (20.8% vs 22%, respectively). When comparing nodular compact calli and fast growing calli, yielding respectively 5% and 100% of “mantled” plantlets, this decrease was up to 4.5% (from 23.2 to 18.7%). An alternative method, the SssI-methylase accepting assay, based on the enzymatic saturation of CG sites with methyl groups, gave convergent results. This work demonstrates that a correlation exists between DNA hypomethylation and the “mantled” somaclonal variation in oil palm. Received: 9 July 1999 / Revision received: 15 October 1999 / Accepted: 26 October     

Evaluation of five promoters for use in transformation of oil palm (Elaeis guineensis Jacq.)

The efficiency of GUS (-Glucuronidase) gene expression in embryogenic callus and young leaflets of mature and seedling palm after microprojectile bombardment with five constructs (pEmuGN, pAHC25, pAct1-F4, pGH24 and pBARGUS) was evaluated to identify the most suitable promoter(s) to use in transformation attempts in oil palm. Expression of the GUS gene driven by theEmu, Ubi1, Act1 35S orAdh1 was assayed, both histochemically and fluorometrically, from a total of 200 plates of tissues in eight independent experiments two days after bombardment. A completely randomized experimental design was used for each experiment, and the data analysed by ANOVA and Duncan's Multiple Range Test. The expression level of GUS driven by theEmu orUbi1 promoters was significantly higher than that of the Act], 35S and Adhl promoters in many experiments, and that of theAdhl was significantly lower than those of the other four promoters. Both histochemical and fluorometric data indicate that in embryogenic callus, the expression of theEmu promoter was higher than that of theUbi1 whereas in young leaflets from mature palm the Ubi1 expression was stronger. The performances of the five promoters were also tested in tobacco callus using a fluorometric GUS assay. The activity of the 35S promoter was highest, and significantly different from that of all the other promoters except theEmu, and that of theAct1 promoter was lowest. These results indicate that either theUbil orEmu promoter should facilitate the expression of desired genes in oil palm and aid in development of an efficient stable transformation system.Abbreviations GUS -Glucuronidase - EC embryogenic callus - YLMP young leaflet from mature palm - YLSP young leaflet from seedling palm - MU methyl umbelliferone - MUG 4-methyl--D-glucuronide - X-glue 5-bromo-4-chloro-3-indoyl-glucuronide - Ubil maize ubiquitin 1 - Actl rice actin 1 - Adh1 maize alcohol dehydrogenase 1 - Emu a recombinant truncated maize alcohol dehydrogenase 1 - ANOVA Analysis of variance - DMRT Duncan's Multiple Range Test Communicated by W A. Parrott     

Application of X-ray microanalysis to cell suspensions of oil palm (Elaeis guineensis Jacq.)

X-ray microanalysis has been used to determine the elemental composition of oil-palm (Elaeis guineesis) cell suspensions without the use of cryoprotectants. Results based on individual cells were gathered over a typical growth cycle of 14 d. During the log phase (5–7 d) there is an increase in the number of cells containing high concentrations of both K (400 mmol kg^-1 dry weight) and P (400 mmol kg^-1 dry weight). Morphologically these cells had thin cell walls and were frequently joined to other cells (two to five cells per clump).     

Cryopreservation of oil palm (Elaeis guineensis Jacq.) somatic embryos involving a desiccation step

Summary The standard cryopreservation process previously developed for oil palm clones using shiny white, finger-like somatic embryos could be applied in some cases to standard cultures. Its efficiency was markedly improved by completing the 7-day pregrowth period on 0.75 M sucrose by an additional dehydration period carried out either by placing the embryos in the air current of the laminar flow cabinet or in an air tight box containing silica gel. This improved process was successfully applied to 7 different clones. It will facilitate the routine uof cryopreservation for oil palm cultures.