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LITERATURE REVIEW

Origin and History of oil Palm It is generally agreed that the Oil Palm (Elaeis guineensis Jacq.) is originated from the equatorial tropical rain forest region of Africa, precisely along the gulf of guinea. It exists in the wild type and cultivated state. The American oil palm, Elaeis oleifera is native to tropical Central America and South America. The main belt runs through the southern latitudes of Cameroon, Cte dIvoire, Ghana, Liberia, Nigeria, Sierra Leone, Togo and into the equatorial region of Angola and the Congo. Oil palm was first illustrated by Nicholaas Jacquin in 1763, hence its name, Elaeis guineensis Jacq (en.wikipedia.org/wiki/Oil_palm#mw-head). During the 14th to 17th centuries some palm fruits were taken to the Americas and from there to the Far East. The plant appears to have thrived better in the Far East, thus providing the largest commercial production of an economic crop far removed from its centre of origin. Oil palms were introduced to Java (Indonesia) by the Dutch in 1848 (Ltschert and Beese 1983) and to Malaysia (then British colony of Malaya) in 1910 by Scotsman William Sime and English banker Henry Darby. The first plantations were mostly established and operated by British plantation owners, such as Sime Darby and Boustead. The large plantation companies remained listed in London until the Malaysian government engineered their "Malaysianisation" throughout the 1960s and 1970s (Stevenson 2006). The cameroons wild oil palm material contains some interesting genotypes which are present into many germplasm in the world. According to Hartley (1988), in Cameroon Germans had identified thin shelled oil palm fruit with high oil content as early as 1902. Later it was known as Tenera type. The material collected by Blaak and Sterling (1996) from some Cameroon regions such as Ekona and Widikum in 1967 have been widely used to develop the majority of oil palm planting seed found today in all the oil palm agroecological area of the world (South East Asia, Africa, South and Central America). According to Chapman et al. (2003), twenty years of breeding using Dami Deli material crossed with Cameroon and Tanzanian selections of African oil palm have led to the development of precocious bearing and cold tolerant oil palms. The oil palm was cultivated in Cameroon since 1907 in the early days of the German colonial era. The first industrial plantations were created in 1910 by Germans in Edea region where the current SPFS Company (Swaziland oil palm Farms Company of Cameroon) is located. In 1929 the Unilever Group created the Pamol Plantations Limited in the British area of the country. Later the British created the Commonwealth Corporation in 1947 which is now known as the Cameroon Development Corporation (CDC).

In the French area of the country another oil palm plantations were created in 1959 by Rivaud Group (Red Land) in Dizangue region and it is currently known as SAFACAM (Forestry and Agricultural African Company of Cameroon). In 1963, the government of Cameroon decided to create the Cameroon Oil Palm Company or SOCAPALM which is currently the largest oil palm company in Cameroon. In 2008, Cameroon covered about 101 500 ha of land plantation of commercial oil palm.

Figure 1. Map showing the extent of oil palm cultivation in 43 oil palm-producing countries in 2006 (FAO 2007). Cited by Koh and Wilcove (2008)

Taxonomy of Oil Palm The scientific classification of oil palm is as follows: Kingdom Phylum/Division Subdivision Class Order Family Subfamily Tribe Subtribe Genus Species : : : : : : : : : : : Plantae Phanerogam/Spermatophyta Angiospermae Monocotyledonae Arecales Arecaceae Arecoideae Cocoseae Elaeidinae Elaeis guineensis and oleifera

Botany of Oil Palm

Oil palm is a diploid (2n=32) oleaginous tropical perennial crop (Dransfield et al. 2005). Elaeis guineensis is a single-stemmed palm which bears a single vegetative shoot apical meristem, which is continuously active producing a new leaf every two weeks in mature palms (Figure 2), under favourable climatic conditions (Adam et al. 2005). Inflorescences are formed throughout the year in acropetal sequence in the axils of subtending leaves of the plant. Separated male and female inflorescences are produced in same palm

plant in alternating cycle of variable duration depending on genetic factors, age and environmental conditions (Corley, 1976).

Figure 2. Adult oil palm (Elaeis guineensis Jacq.) tree

The Foliage System of Oil Palm At the mature stage, E. guineensis Jacq. is constituted with a large crown of 30 to 45 palms measuring 5 to 9 meters long topping the single cylindrical pseudo-trunk. Every year, about 20 to 26 new pinnate leaves are produced and each mature leaf bears 250 to 300 leaflets. The leaf bases are persistent for years, and prominent leaf scars are arranged spirally on the trunk of mature palms where bases have fallen. The leaflets cover the distal 2/3 of the leaf, and the lower 1/3 is spined with spines increasing in length acropetally. The stem or pseudo-trunk can extend a rate of 30 to 60 cm/year between the ages of 6 to 15 depend on hereditary and environmental factors. It functions as a supporting, vascular and storage organ. Generally, the height of trees determines the exploitation times of oil palm plantations. Thus, palm plantations are exploited up to 25 to 30 years when the trees height is between 12 to 15 meters. Above of this, the harvest of fresh fruit bunches becomes difficult and replanting should be required.

The Roots System of Oil Palm The root architecture is a fundamental aspect of plant productivity through its functional importance in the efficient acquisition of soil resources (water and nutrients

uptake). The different morphological types of oil palm roots have been distinguished according to their development pattern and state of differentiation. As all the monocot, the root system of E. guineensis is fasciculata. The oil palm has an adventitious root system; with primary roots generally about 6-10 mm in diameter, originating from the base of the trunk and either spreading horizontally or descending at varying angles into the soil. The primary roots bear secondary roots, of about 2-4 mm in diameter. Tertiary roots, about 0.7-1.2 mm in diameter, branch out from the secondary roots, which in turn bear the quaternary roots. Quaternary roots are unlignified, about 0.1-0.3 mm in diameter and 1-4 mm long and they are often assumed to be the main absorbing roots (Corley et al., 1976). The total length of tertiary and quaternary roots in the soil is the most important root characteristic as they are the absorbing roots that affect fertilizer use efficiency. Most of the root biomass is found within 1m of the soil surface, but tertiary and quaternary roots are found mostly in the upper 30 cm from the soil surface. Primary roots can grow up to 20 m away from the base of the palm and some primary roots could penetrate below the water table at 90 cm from the surface (Ng et al., 2003). The distribution of roots depends largely on the nature of the soil. Oil palm being a monocot needs a friable soil for root branching (Zuraidah et al. 2010).

The Reproductive System of Oil Palm As monoecious plant, male and female flowers of oil palm occur separately on the same tree. The male and female inflorescences are produced distinctly and alternatively on the same plant. An inflorescence is initiated in the axil of every leaf. An inflorescence may bear about 250 spikelets. Each female inflorescence (Figure 3B) can have about 12 to 30 flowers. Sometimes hermaphrodites flowers can also occur but in rare cases. The female flower is receptive for pollination between 36 to 48 hours. The male inflorescence (Figure 3A) contains about 100 000 flowers. Most of the pollen released is shed between 2 to 4 days after opening. The pollen remains viable for 6 days after release. The insufficient supply of water will stimulate the plant to produce mostly male flowers. Irrigation and fertilization taken to increase the flower ratio from male to female will have positive effect only between 22 to 40 months later.

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Figure 3. A. Male inflorescence, and B. Female inflorescence of oil palm. In the nature, the pollination is mainly entomophily (by insects) but can also be done by the wind and the rain. The main pollinator of the oil palm is the insect from the genus Elaeidobius and 14 species were found to be associated to the male and female inflorescences. The most common one is Elaeidobius kamerunicus, which was introduced in Asia, South and Central America. Its introduction led to an increase in yields of more than 35%. Upon pollination at the anthesis stage, the female inflorescence may develop and give rise to a fruit bunch 22 to 26 weeks later. In the field, fruit bunches (Figure 4) which are as compact and ovoid mass spiked bearing spines can be found on the oil palm 2 to 3 years after planting. The fruit bunches become heavier as the palms get older. On 10 years old palms, the bunch weights can be ranged from 10 to 50 kg for a total of 500 to 4 000 fruits per bunch. The fruit is a sessile drupe generally of ovoid shape measuring 2 to 5 cm length and weighing from 3 to 30g. The oil palm fruit consists of the pericarp which includes the outer and smooth exocarp, the fibrous and fleshy mesocarp or pulp rich in palm oil, the hard endocarp or shell protecting the seed and finally the kernel or seed. Inside the shell is the endosperm or kernel. The endosperm contains large amount of fats and carbohydrates on which the seedling will be entirely dependent after germination. The oil palm produces two main vegetable oils, namely palm oil extracted from the mesocarp of the fruit and palm kernel oil extracted from seed.

Immature

Mature

Figure 4. Mature and Immature Fresh fruit bunches of oil palm

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Commonly three fruit forms or varieties of oil palm can be identified based on the thickness of the shell criterion (Figure 5). The Dura has a shell thickness between 2 to 8 mm, while the Pisifera has no shell. The shell thickness of the hybrid Tenera is between 0.2 to 2 mm, the fleshy mesocarp of Dura yields between 15 to 17% oil while in Tenera yields between 21 to 23% oil and Pisifera more than 23% oil. According to commercial purpose, Pisifera is not cultivated on large scale because of its ability to fruits abortion and thereby virtual empty bunches production. Tenera which is a hybrid of Dura and Pisifera varieties remains the commercial variety with 60 to 96% of mesocarp. The hybrid produces more fruit bunches than Dura.

B A C

Figure 5. Cross section of the different varieties of oil palm ( E. guineensis Jacq.). A. Dura palm fruit with thick shell, B. Tenera palm fruit with less thick shell, C. Pisifera palm fruit with no shell

Climatic and Soil Requirements of Oil Palm Oil palm culture is done well in low altitude (less than 500 m above sea level), 15 from the equator in the humid tropics. Evenly distributed rainfall of 1,800 to 2,000 mm/year, but will tolerate rainfall up to 5,000 mm/year, provided the soil is properly drained. Oil palm is sensitive to poor drainage and drought. Potential yield is reduced where there are more than three consecutive months with less than 100 mm rainfall per month. Irrigation may increase economic returns in areas with pronounced dry periods. More than 2,000 sunshine hours (i.e., low cloud cover during daytime) are required. It is adapted to a range of soil types. Tolerates low pH, but does not thrive at very high pH (greater than 7.5). Soil must be free draining. The table below shows the suitable and extreme values of climate and soil that are required by oil palm.

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Table 1. Agro ecological factors affecting growth and production of oil palm Characteristic Rainfall (mm) Temperature (oC) Water deficit (mm)
Solar irradiation Dry season (mths) Wind (m/s)

Good 2500 25-30 0-150 13-15

none 5-9 5-6 0-4 Never Good

Suitability class Moderate 1450-1700 20-22 150-250 9-11

1-2 10-15 4.5-5 4-12 Minor Poor

Severe 1250-1450 10-20 250-400 7-9

2-4 15-20 4-4.5 or 6.5-7 12-23 Moderate Very excessive

Very severe <1250 <16 >400 <7

>4 >20 <4 or >7 >30 Severe flooding Very poor

pH Slope (%) Flooding Drainage class

PLANT TISSUE CULTURE

History of Plant Tissue Culture

Many authors have put forward various arguments on the origin of plant tissue culture. Rehwald (1927) is thought to have been the first person who has obtained undifferentiated callus tissue in sterile culture. But according to most scientists, the real scientific work on tissue culture was done in 1934 by P.R. White in the USA (United States) and in the same year, by Gautheret in France, using respectively tomato plant and the cambium of several tree species (Acer pseudoplatanus, Salix caprea, sambucus) as sources of explant. Before that period, in 1924, callus culture of carrots ( Daucus carota) was reported by two physicians, R. Blumenthal and P. Meyer in the pathological implication studies. In 1934, the plant hormone or auxin named indole-3-acetic acid (IAA) was identified the first time by F. Kgl, A.J. Haagen-Smit, and H. Erxleben. In 1955 C.O Miller has discovered kinetin, which is a plant growth regulator known as cytokinin. Plant tissue culture is a technique or a process by which small organs or pieces of tissue of plants (explants) are grown in aseptic conditions within glass or clear plastic vessels. These are usually termed in vitro techniques or propagation. It is also called micropropagation because miniature shoots and plantlets are initially derived. The natural capability of plants to multiply by asexual means is the basis for multiplication in vitro. Tissue culture was first used

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on a large scale by the orchid industry in the 1950s. Later, it became clear that any plant would respond to tissue culture as long as the right formula and the right processes were developed for its culture. The process of tissue culturing plant the explant stage to the final stage of plantlet requires four basic stages namely: establishment or initiation of explant (Stage I), multiplication of cells (Stage II), rooting of shoots (Stage III), and acclimatization or hardening off (Stage IV).

Explants

The kind of explant chosen, its size, age and the manner in which it is cultured, can all affect whether tissue culture can be successfully initiated and whether morphogenesis can be induced. The juvenile form had pre-existing competent cells that were able to respond to auxin and become determined to form organs. However, the adult form appeared to lack cells with pre-existing competence to form organs, but competence was acquired by some callus cells once they had been initiated. Explants taken from mature shoots are frequently more liable than juvenile material to suffer necrosis, especially when surface disinfested and placed in culture (Hanus and Rohr, 1987). For tissue culture, juvenile explants are usually more readily established in vitro and grow and proliferate at a more rapid rate than adult material. This is particularly true with tree species where micropropagation of adult material is often difficult (Edwin et al., 2008).

Medium of Tissue Culture

The success of plant tissue culture technique is greatly influenced by the nature of the culture medium. The medium is the substrate for the started explant and it refers to an aseptic environment of the mixture of a correct concentration and balanced chemical diet compounds to form a nutrient-rich gel or liquid for growing cultures, whether cells, organs, or plantlets. Plant tissue culture media are therefore made up from macronutrients salts which provide the six major elements are N, P, K, Ca, Mg, and S, contributing in the growth of higher plants; micronutrient salts are Fe, Mn, Zn, B, Cu, Co and Mo which are components of plant cell proteins of metabolic and physiological importance: chlorophyll synthesis and chloroplast function (Sundqvist et al., 1980); vitamin; amino acids and other nitrogen supplements; sugar, buffers; solidifying agent; and growth regulators. The medium pH must be such that it will not disrupt the function of plant cell membranes or the buffer pH of cytoplasm, and the

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gelling efficiency of agar. The initial pH can often be selected to ensure the integrity of the medium and the most rapid rate of culture growth (Edwin and Sherrington, 1984). However a supply of reduced nitrogen is generally necessary for embryogenesis to occur in callus or suspension cultures.

Plant Growth Regulators

Endogenous compounds occur naturally within plant tissues. They have a regulatory role rather than nutritional activity in growth and development. These compounds are always active at very low concentrations and known as plant growth substances or plant hormones. Otherwise synthetic chemicals with similar physiological activities can be found and generally supplied in the medium for growth and morphogenesis in vitro. They are termed plant growth regulators (exogenous). Some recognized plant growth substances classes include such chemical compounds as auxins, cytokinins, gibberellins, ethylene and abscisins. The first two compound classes are by far the most important for regulating growth and morphogenesis in plant tissue and organ cultures. Auxins are phytohormones very widely used in micropropagation medium to promote the growth of callus, cell suspensions or organs and to regulate morphogenesis, mainly when combined with cytokinins. They influence cell enlargement, root initiation, and adventitious bud formation. The potential levels of endogenous auxin in the explant tissues are found to depend on the mother plant (donor plant growth state, growth conditions and season of sampling) from which they were collected. Therefore, the natural auxin will vary between cell strains of the same origin. Meristematic cells are referred to be particular active sites for the biosynthesis and/or the release of natural growth factors (Clare and Collin, 1974). The

commonly auxins used in tissue culture consist of NAA (1-naphthaleneacetic acid); 2,4-D (2,4-dichlorophenoxyacetic acid); IAA (indole-3-acetic acid); IBA (indole-3-butyric acid); Dicamba (3,6-dichloro-0-anisic acid or 3,6-dichloro-2-methoxybenzoic acid); Picloram (4Amino-3,5,6-trichloro-2-pyridinecarboxylic acid ). Formerly called kinins, cytokinins are growth regulators that are required in tissue culture media for cell division, shoot multiplication, and axillary bud proliferation. In addition, they help delay senescence, and influence the transport of auxins. Cytokinins which are commonly used in tissue culture media include Kinetin (6-furfurylaminopurine); Zeatin (6-[4-hydroxy-3-methylbut-2-enlyamino]purine); BA or BAP (6-benzylaminopurine); 2iP (N6-[2-isopentenyl]adenine or 6-[,-dimethylallylamino]purine).

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Zygotic Embryo Culture

Zygotic embryo culture is the aseptic isolation and growth of sexually produced embryo in vitro with the objective of obtaining viable plants. Two principle types of embryo culture are concerned: a) culture of immature embryos, where isolated embryos are originated from unripe or hybrid seeds which failed to germinate are used to be grown; b) culture of mature embryos, where mature embryos are excised from ripe seeds and culture mainly to avoid inhibition in the for germination. An embryo callus possesses a high regenerative capacity compared to those derived from mature organs such as the leaf, stem and root. The age of embryo considerably influences the regenerative ability of plant calluses. The most useful and popular application of embryo culture, has been to raise rare hybrids by rescuing embryos of incompatible crosses. In fact, poor or abnormal development of the endosperm causes starvation and eventual abortion. Isolation of hybrid embryos before abortion and their in vitro culture may circumvent these strong post-zygotic barrier. Plant embryo culture is an invaluable breeding technique as far as it is possible to synthesize hybrids from incompatible crosses. The regenerative potential is an essential prerequisite in non conventional method of plant genetic manipulation. Because of their juvenile nature, embryos have a high potential for regeneration and hence may be used for in vitro clonal propagation (Razdan, 2003).

Ovary Culture

The culture of unfertilized ovaries to obtain haploid plants from egg cell or other haploid cells of the embryo sac (synergid, antipodal) is called ovary culture, and the process is termed as gynogenesis (in vitro parthenogenesis). The rate of success varies considerably with species and is markedly influenced by explant genotype so that same cultivars do not respond at all. As in anther culture, gynogenesis may occur either via embryogenesis or through plantlet regeneration from callus. The frequency of responding ovaries (1-5%) and the number of plantlets per ovary (1-2) is quite low (Yunbi, 2010).

Tissue Culture Problems and Preventions

Genetic Changes in Culture. Selecting a suitable source of explant material is essential to the success of tissue culture process. Culture and subcultures should be initiated with clean

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and disease-free material from genetically uniform plant. Basically in mother plant, vegetative parts are composed of cells that all have the same genetic potential (genotype), however sometimes, intact plants may be composed of cells with different genotypes in separate layers or segments, or occurring in a random mosaic fashion. This occasionally genetic variable of mother plant tissues can result to genetic changes in culture. Thus, care must be taken during explant sampling to be sure that origin explants are not taken from plant tissue likely to be endopolyploid. Subsequent exposure to high levels of growth hormones such as 2,4-D should be also avoided as far as possible. Besides of this, polyploid or aneuploid cells, or cells which contain structurally altered chromosomes may arise directly in culture from callus or suspension cultures (atypical or abnormal plants). But the extent and occurrence of genetic disturbance depend on the genetic constitution of the cells which are cultured (Edwin and Sherrington, 1984).

Blackening or Browning. Plants naturally contain tannin substances or hydroxyphenol which affect growth. Explants from those plants frequently turn brown or black shortly after isolation and growth is inhibited and the tissue usually dies. Young tissues are often less liable to browning on excision than older one. Some pretreatments of plant material can be used to remove phenolic compounds such as: (a) leaching or soaking in running water during 24 hours or in sterile water for 2-3 hours; (b) absorption by activated charcoal in the medium; (c) absorption by polyvinylpyro-lidone (PVP); (d) modifying redox potential by using reducing agents (anti-oxidant); (e) inhibiting the action of phenol oxidase enzymes (darkness, pH) (Edwin F.G and Sherrington P.D, 1984).

Embryogenesis The earlier work on somatic embryogenesis was conducted on carrot ( Daucus carota). The initiation and the development of embryo cells from somatic tissues in plant tissue culture was first observed by Steward et al. (1958) and Reinert (1958, 1959), using Daucus carota explant. The presence of auxin compound or auxin-like substance in the culture medium promote the embryos initiation, while reducing its concentration or the complete absence of that hormone allows the maturation of embryo. Sharp et al. (1980) are described two pathways of somatic embryogenesis. The first is direct embryogenesis where embryos can be initiated directly from tissue in the absence of callus proliferation. This occurs through preembryogenic determined cells. In this case cells are already committed to embryogenic

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development and need only to be released. The second is indirect embryogenesis where cell proliferation or callus is required. This occurs in differentiated non-embryogenic cells or induced embryogenic determined cells. Therefore, under certain cultural and environmental conditions, it is possible to regulate initiation and development of embryogenic cells. In addition, the frequency of response will vary considerably with the genotypes of the species used. However the plants from monocotyledonae class have proven difficult to grow in culture and regenerate somatic embryos. Tissue Culture Field in Plant Breeding Tissue culture is a field of many facets. It is the ever-ready tool for breeders who hybridize plants by either sexual or asexual means. Cell and tissue culture are the innovative breeding techniques applied to meet the increasing need for improved crop varieties. Tissue culture techniques can shorten the time and can lessen the labor and space requirements needed to produce a new variety. It is a clean and rapid mean for genetic engineers to grow material for identifying and manipulating genes or to transfer interesting individual characteristics from one plant to another. Cell and tissue culture and recombinant DNA technology constitutes an important aspect of plant biotechnology (Chawla, 2002). Generally, tissue culture plays an important role in wide array of fields, such as molecular biology, hybrid development, genetic engineering, pesticide testing, botany, chemistry, physics, and food science (Lydiane Kyte and John Kleyn, 1983). Pathogen-free plants from tissue culture have opened the door to a freer exchange of plants between states and countries. They have gained acceptance in world trade because of the virtual elimination of the disease introduction dangers which can occur. Most tissue cultured plants are true to type, more vigorous, more disease resistant, and disease free. The free exchange of tissue cultures will have a significant impact on global food problems by allowing for more improved cultivars to be obtained more rapidly to growers worldwide. Plant tissue culture involves asexual methods of propagation and its primary goal is crop improvement. The success of many in vitro selections and genetic manipulation techniques in higher plants depends on the success of in vitro plant regeneration. Crop breeding and rDNA technology require the widespread use of reliable true to type propagation and better regeneration methods. The tissue culture technique is useful for in vitro production of haploids, in which plants with gametophytic number of chromosomes (single set) in their sporophytes are obtained. Haploids are mostly obtained from anther or pollen culture, ovule

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or ovary culture and chromosome elimination after inter specific hybridization. Double haploids or homozygous plants can then be obtained by colchicines treatment, fusion of pollen nuclei and endomitosis. The interest in haploids by plant breeder stems largely from their considerable potential in plant breeding, especially for the quick production of homozygous plants and isogenic lines that would result in the early release of crop varieties. This would also facilitate work in inducing and detecting desirable mutations, transformations, and biochemical genetics. In other ways, somatic cell fusion (protoplast) under in vitro condition leads to the formation of cell hybrids which are very important for somatic cell genetics and crop improvement in the fields of plant breeding program. In addition, as the genetic variability is an essential component of any breeding program designed to improve the characteristics of crop plants, somaclonal variation through plant cell culture has been promoted as one of the potential sources of useful genetic variation. In another words, the genetic differences between plants regenerated from callus are sufficient to have attracted the interest of plant breeders as a new source of selectable variability. Thus in vitro culture of higher plants can be used for selection of mutants (in vitro selection) (Evans et al.,1983 ; Chawla, 2002).

Oil Palm Genetic Improvement

The high world demand for oils and fats due to their uses as renewable bio-energy supplies and crucial source of food and without forgetting the environmental effect reduction need of oil palm have boosted the oil palm improvement sector. It was estimated about 155 million tons in 2007 (Oil world annual report 2008). Based on current demand for oil palm seeds in the world, Zamzuri et al., (1999) estimated that there is a ready market for more than 100 million tissue culture plantlets annually. The programme of crop improvement through the utilization of traditional breeding and selection methods, the development and benefits of tissue culture techniques and ongoing efforts to apply molecular and genetic engineering techniques to improve and modify oil composition, are continuously applied. The four African Elaeis guineensis palms brought over by the Dutch in 1848, and planted in Buitenzorg Botanical Garden (now Bogor) Indonesia laid the foundation for the oil palm industry in Indonesia and Malaysia. From these, the Deli dura palms with unique and favourable fruit qualities were developed. In the conventional breeding methods, the Deli dura population (group A) with small number of large bunches and the African Pisifera and

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Tenera palms (group B) which consists of large number of small bunches, are widely utilized for seed production and in genetic improvement programmes. Oil palm improvement is carried out mainly in South East Asia by Indonesia and Malaysia. In Africa some research centre of oil palm include CEREPAH of La-Dibamba in Cameroon, CNRA of La-Me in Ivory Coast, NIFOR in Nigeria and Benin. Apart from raising the total yield of fresh fruit bunches (FFB), breeding and selection also focus on achieving high FFB oil and kernel content. The quality of the oil in terms of a high level of unsaturation [high iodine value (I.V)] and minor but important constituents such as vitamin E and carotenoids, is also being selected for. Vegetative characters are also taken into account where reduced rates of trunk extension and long bunch stalks are desirable attributes to facilitate harvesting while compact palms may allow higher planting densities of up to 180 palms/hectare (Basri et al., 2003). Elaeis oleifera, an oil palm species endemic to South and Central America readily hybridizes with Elaeis guineensis. This American species offers several desirable traits including slow height increment, high unsaturation and resistance to disease such as Fusarium wilt, which can be introgressed into the economically important Elaeis guineensis. Generally the objective of plant breeding in oil palm is to create genotypes with the maximum potential for oil and kernel production per unit of area, and some morphological characters, the tolerance to abiotic and biotic stress. Breure & Bos (1992) and Breure & Verdooren (1995) described the strategy for selecting superior materials, which involves three steps: 1) phenotypic selection of superior palms by the way of the reciprocal recurrent selection (RRS) method in which intra population crossing of few parents with favorable genes and constant heterogeneity is done in each population follow by inter population crossing of the best selected parents; 2) selection based on general combining ability (GCA) by means of progeny tests; and 3) the evaluation of the best families, carrying out crossings between them to exploit GCA and specific combining ability (SCA). The selected Dura family is crossed with a superior Pisifera (tester). In addition, backcrosses are also used in oil palm to introgress specific characteristics from a donor parent into a recipient parent. Tissue Culture and Oil Palm Improvement The earliest reports of successful vegetative propagation of oil palm by tissue culture were in the mid 1970s (Jones, 1974; Rabechault and Martin, 1976). Currently, about 20 oil palm laboratories are functionnal throughout the world with capacity ranging from 10,000

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200,000 plantlets per year (Zamzuri et al., 1999). As compared to seed production, tissue culture of oil palm offers several advantages (Sogeke, 1998). It allows rapid multiplication of uniform planting materials with desired characteristics. This enables improvement of planting materials using existing individuals which have all or most of the desired qualities such as good oil yield and composition, slow vertical growth and disease resistance. Additionally, it also opens new avenues for producing novel planting materials via genetic engineering, because tissue culture is the means for regeneration of tissues transformed with genes for traits of interest. The selection of the donor palm (ortet) is based on a range of characteristics such as yield, extraction rates, height and disease or drought tolerance. These palms are graded and a list submitted to the tissue culture laboratory for propagation (clonal propagation). Plant tissue aims to clone a minimum of 50 grade 1 palms, 40 grade 2 and 30 grade 3 suitable for planting in the field. The most responsive parts of the oil palm in regard to tissue culture are those areas which are meristematic (rapidly dividing) and young. Ideally the explant should provide large numbers of starting tissues and have little or no contamination. Oil palm tissue culture is employed both as a means for producing good tenera palms for commercial planting and to multiply good parents (both dura and pisifera) for seed production. It is also practised to expedite the exploitation of progenies from interspecific E. oleifera X E. guineensis crosses. The main problem of oil palm breeding is to obtain oil palm homozygous true breeding line due to its long breeding cycle. By conventional inbreeding and backcrossing it is possible to obtain pure lines, but it is a long and cumbersome process which needs 6-8 rounds of controlled selfing (30 years) of the oil palm in order to get homozygote inbred. However, homozygous plants can be obtained in a relatively short time (within a year) by the production of haploids and by doubling their chromosomes; this is done by Sumatra Bioscience which produces about 5 oil palm haploids per day. Tissue culture laboratories are linked to an effective oil palm breeding and improvement programme to ensure supply of desired explants. Ortets selected are supported by at least four years of field data showing good performance on oil yield, vegetative characteristics such as low height increment and physiological traits such as bunch index and oil characteristics (Rohani et al., 2000). Based on field performance data from various sources an improvement of oil yield between 20% to 30% over seedling planting materials is achievable by clonal materials (Soh et al.,2001). There are indications that selection for resistance to major oil palm diseases will be easier based on differences in susceptibility shown by clones to Ganoderma, Fusarium and blast (Purand-Gasselin et al., 1999). The difficulty lies in achieving true-to type reproduction of plants selected as ortets,

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especially with the incidence of mantled abnormality. The percentage of abnormality in the field has generally been maintained at a tolerable level of less than 5% (Maheran et al., 1995). Prudent selection of good ramets at both in vitro and nursery stage is practised to reduce abnormality level. Maheran et al. (1995) reported that clones appearing normal at both stages gave a low level of abnormality in the field (about 2.2%). It was estimated that the initial investment on clonal materials will be covered in the sixth year (Zamzuri et al., 1999), after which the returns from clonal materials will be much higher than conventional Dura X Pisifera planting materials due to their higher productivity.