WO2013089557A1 - Methods for obtaining high-yielding oil palm plants - Google Patents

Methods for obtaining high-yielding oil palm plants Download PDF

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Publication number
WO2013089557A1
WO2013089557A1 PCT/MY2012/000241 MY2012000241W WO2013089557A1 WO 2013089557 A1 WO2013089557 A1 WO 2013089557A1 MY 2012000241 W MY2012000241 W MY 2012000241W WO 2013089557 A1 WO2013089557 A1 WO 2013089557A1
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Prior art keywords
oil palm
fruit
level
palm plant
weeks
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PCT/MY2012/000241
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English (en)
French (fr)
Inventor
Huey Fang TEH
Ping Li MAY HONG
Bee Keat NEOH
Yoke Sum JAIME LOW
Binti Ithnin NALISHA
Lee Mei THERESA NG
Yin Mee THANG
David Ross APPLETON
Yusof@Hassan HIRZUN BIN MOHD
A/L Kulaveerasingam HARIKRISHNA
Mohamed MOHAIMI
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Sime Darby Malaysia Berhad
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Priority to SG11201403224YA priority Critical patent/SG11201403224YA/en
Priority to BR112014014741A priority patent/BR112014014741A2/pt
Publication of WO2013089557A1 publication Critical patent/WO2013089557A1/en
Priority to CR20140282A priority patent/CR20140282A/es

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/08—Fruits

Definitions

  • This application relates to methods for obtaining high-yielding plants, and more particularly to methods for obtaining oil palm plants that are high-yielding with respect to producing palm oil.
  • the African oil palm Elaeis guineensis Jacq. is an important oil-food crop.
  • Oil palm plants are monoecious, i.e. single plants produce both male and female flowers, and are characterized by alternating series of male and female inflorescences.
  • the male inflorescence is made up of numerous spikelets, and can bear well over 100,000 flowers.
  • Oil palm is naturally cross-pollinated by insects and wind.
  • the female inflorescence is a spadix which contains several thousands of flowers borne on thorny spikelets. A bunch carries 500 to 4,000 fruits.
  • the oil palm fruit is a sessile drupe that is spherical to ovoid or elongated in shape and is composed of an exocarp, a mesocarp containing palm oil, and an endocarp surrounding a kernel.
  • Oil palm is important both because of its high yield and because of the high quality of its oil.
  • yield oil palm is the highest yielding oil-food crop, with a recent average yield of 3.67 tonnes per hectare per year and with best progenies known to produce about 10 tonnes per hectare per year.
  • Oil palm is also the most efficient plant known for harnessing the energy of sunlight for producing oil.
  • quality oil palm is cultivated for both palm oil, which is produced in the mesocarp, and palm kernel oil, which is produced in the kernel. Palm oil in particular is a balanced oil, having almost equal proportions of saturated fatty acids ( ⁇ 55% including 45% of palmitic acid) and unsaturated fatty acids ( ⁇ 45%), and it includes beta carotene.
  • the palm kernel oil is more saturated than the mesocarp oil. Both are low in free fatty acids.
  • the current combined output of palm oil and palm kernel oil is about 50 million tonnes per year, and demand is expected to increase substantially in the future with increasing global population and per capita consumption of oils and fats.
  • oil palm is the highest yielding oil-food crop, current oil palm crops produce well below their theoretical maximum.
  • conventional methods for identifying potential high-yielding palms for use in crosses to generate progeny with higher yields require cultivation of palms and measurement of production of oil thereby over the course of many years, which is both time and labor intensive.
  • Transgenic approaches offer potential solutions to the general problem of the need to increase plant yields.
  • transgenic modification of crops such as soy and corn by the introduction of pest resistance genes derived from other organisms is now well known as a means for increasing crop yields.
  • methods for increasing plant yields by increasing or generating in the plant activities of particular proteins have also been disclosed, for example by Schon et al., WO 2010/046221.
  • transgenic modification of crops raises potential concerns regarding unintended detrimental effects on individuals and ecosystems.
  • Metabolomics which encompasses the study of the metabolite complement of a cell, tissue, or organism, also offers potential solutions to the general problem of increasing plant yields.
  • metabolic profiling corresponding to non-biased screening of biological samples for changes of metabolite levels relative to control samples, provides a means to determine whether and to what extent the presence and levels of particular metabolites may differ between biological samples subjected to particular treatments or at particular points in development or time, as discussed for example in J. Kopka, Gas Chromatography Mass Spectrometry, in 57 PLANT
  • a method for obtaining a high-yielding oil palm plant comprises determining the level of a metabolite in mesocarp tissue of a fruit of a parental oil palm plant 11 to 23 weeks after pollination thereof.
  • the metabolite is selected from the group consisting of 2-aminobutyric acid, 2-hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine,
  • the method also comprises determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 1 1 to 23 weeks after pollination thereof.
  • the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the method also comprises selecting progeny of the parental oil palm plant based on the difference to obtain the high- yielding oil palm plant.
  • the method comprises determining the level of a metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 23 weeks after pollination thereof.
  • the metabolite is selected from the group consisting of 2-aminobutyric acid, 2-hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine, cyclohexylamine, cytidine, gamma-aminobutyric acid, glyoxylic acid, guanosine, homo- L-arginine, glutamine, indole-3-acetaldehyde, lysine, myo-inositol-phosphate, myo- inositol-2 -phosphate, nicotinamide, ⁇ - ⁇ -methylarginine, octylamine,
  • the method also comprises determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 23 weeks after pollination thereof.
  • the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 15% higher than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the method also comprises predicting the oil yield of the test oil palm plant based on the difference.
  • kits for obtaining a high-yielding oil palm plant or predicting oil yield of a test oil palm plant comprises a reagent for quantitative detection of a metabolite.
  • the metabolite is selected from the group consisting of 2-aminobutyric acid, 2-hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine, cyclohexylamine, cytidine, gamma-aminobutyric acid, glyoxylic acid, guanosine, homo-L-arginine, glutamine, indole-3-acetaldehyde, lysine, myo-inositol-phosphate, myo-inositol-2 -phosphate, nicotinamide, co-N-methylarginine, octylamine, ophthalmic acid, serine, sper
  • the kit also comprises an extract of a mesocarp tissue of a fruit of a reference oil palm plant 11 to 23 weeks after pollination thereof.
  • the kit also comprises instructions indicating use of the reagent for determining whether there is a difference between the level of the metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 1 1 to 23 weeks after pollination thereof, and the level of the metabolite in the extract of the mesocarp tissue of the fruit of the reference oil palm plant.
  • the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental or test oil palm plant is at least 15% higher than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the method comprises determining the level of a metabolite in mesocarp tissue of a fruit of a parental oil palm plant 1 1 to 23 weeks after pollination thereof.
  • the metabolite is selected from the group consisting of 1-methyladenosine, 4-aminophenylsulfone, 4- guanidinobutyric acid, 6,8-thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N- acetylhistidine, N-methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine,
  • the method also comprises determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 23 weeks after pollination thereof.
  • the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the method also comprises selecting progeny of the parental oil palm plant based on the difference to obtain the high-yielding oil palm plant.
  • the method comprises determining the level of a metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 23 weeks after pollination thereof.
  • the metabolite is selected from the group consisting of 1-methyladenosine, 4-aminophenylsulfone, 4- guanidinobutyric acid, 6,8-thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazoIe-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N- acetylhistidine, N-methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tarta
  • the method also comprises determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 1 1 to 23 weeks after pollination thereof.
  • the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the method also comprises predicting the oil yield of the test oil palm plant based on the difference.
  • kits for obtaining a high- yielding oil palm plant or predicting oil yield of a test oil palm plant comprises a reagent for quantitative detection of a metabolite.
  • the metabolite is selected from the group consisting of 1 - methyladenosine, 4-aminophenylsulfone, 4-guanidinobutyric acid, 6,8-thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N-acetylhistidine, N-methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tartaric acid, tetrahydrouridine,
  • the kit also comprises an extract of a mesocarp tissue of a fruit of a reference oil palm plant 11 to 23 weeks after pollination thereof.
  • the kit also comprises instructions indicating use of the reagent for determining whether there is a difference between the level of the metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 11 to 23 weeks after pollination thereof, and the level of the metabolite in the extract of the mesocarp tissue of the fruit of the reference oil palm plant.
  • the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental or test oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the method comprises determining the level of a metabolite in mesocarp tissue of a fruit of a parental oil palm plant 11 to 23 weeks after pollination thereof.
  • the metabolite is selected from the group consisting of adenosine triphosphate and uridine triphosphate.
  • the method also comprises determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 23 weeks after pollination thereof.
  • the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the method also comprises selecting progeny of the parental oil palm plant based on the difference to obtain the high-yielding oil palm plant.
  • the method comprises determining the level of a metabolite in mesocarp tissue of a fruit of a test oil palm plant 1 1 to 23 weeks after pollination thereof.
  • the metabolite is selected from the group consisting of adenosine triphosphate and uridine triphosphate.
  • the method also comprises determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 23 weeks after pollination thereof.
  • the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the method also comprises predicting the oil yield of the test oil palm plant based on the difference.
  • kits for obtaining a high-yielding oil palm plant or predicting oil yield of a test oil palm plant comprises a reagent for quantitative detection of a metabolite.
  • the metabolite is selected from the group consisting of adenosine triphosphate and uridine triphosphate.
  • the kit also comprises an extract of a mesocarp tissue of a fruit of a reference oil palm plant 11 to 23 weeks after pollination thereof.
  • the kit also comprises instructions indicating use of the reagent for determining whether there is a difference between the level of the metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 11 to 23 weeks after pollination thereof, and the level of the metabolite in the extract of the mesocarp tissue of the fruit of the reference oil palm plant.
  • the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental or test oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • FIG. 1 is a graph of results of principal component analysis with respect to metabolites of mesocarp tissue of fruits of five high-yielding oil palm plants and five low-yielding oil palm plants at week 16 post-pollination.
  • FIG. 2 is a graph of ion chromatograms of spermine as detected in samples prepared from mesocarp tissue of fruits of high-yielding oil palm plants and low-yielding oil palm plants at week 16 post-pollination.
  • FIG. 3 is a graph of results of Orthogonal Partial Least Square-Discrimination Analysis, i.e. supervised analysis, with respect to metabolites of mesocarp tissue of fruits of five high-yielding oil palm plants and five low-yielding oil palm plants at week 16 post-pollination.
  • FIG. 4 provides a heat map indicating relative levels of various metabolites of the citric acid cycle metabolic pathway, as determined for mesocarp tissue of fruits of high- versus low-yielding oil palm plants, at weeks 12, 14, 16, 18, 20, and 22 post-pollination (increasing darkness of shading indicates increasing relative level of metabolite).
  • FIG. 5 is a graph of relative concentration versus time (weeks 12 to 22) post- pollination for various metabolites of the citric acid cycle metabolic pathway, as determined for mesocarp tissue of fruits of high- versus low-yielding oil palm plants.
  • FIG. 6A provides a heat map indicating relative levels of various metabolites of amino acid metabolic pathways, as determined for mesocarp tissue of fruits of high- yielding oil palm plants, at weeks 12, 14, 16, 18, 20, and 22 post-pollination (increasing darkness of shading indicates increasing relative level of metabolite).
  • FIG. 6B provides a heat map indicating relative levels of various metabolites of amino acid metabolic pathways, as determined for mesocarp tissue of fruits of low- yielding oil palm plants, at weeks 12, 14, 16, 18, 20, and 22 post-pollination (increasing darkness of shading indicates increasing relative level of metabolite).
  • FIG. 7 is a graph of relative concentration versus time (weeks 12 to 22) post- pollination for various metabolites of amino acid metabolic pathways, as determined for mesocarp tissue of fruits of high- versus low- yielding oil palm plants.
  • FIG. 8 provides a heat map indicating relative levels of various metabolites of purine and pyrimidine metabolic pathways, as determined for mesocarp tissue of fruits of high- versus low-yielding oil palm plants, at weeks 12, 14, 16, 18, 20, and 22 post- pollination (increasing darkness of shading indicates increasing relative level of metabolite).
  • FIG. 9 is a graph of relative concentration versus time (weeks 12 to 22) post- pollination for various metabolites of purine and pyrimidine metabolic pathways, as determined for mesocarp tissue of fruits of high- versus low-yielding oil palm plants.
  • FIG. 10 provides a heat map indicating relative levels of various metabolites of oxidative phosphorylation metabolic pathways, i.e. electron transport chain and ATP synthesis, as determined for mesocarp tissue of fruits of high- versus low-yielding oil palm plants, at weeks 12, 14, 16, 18, 20, and 22 post-pollination (increasing darkness of shading indicates increasing relative level of metabolite).
  • oxidative phosphorylation metabolic pathways i.e. electron transport chain and ATP synthesis
  • FIG. 11 is a graph of relative concentration versus time (weeks 12 to 22) post- pollination for various metabolites of oxidative phosphorylation metabolic pathways, i.e. electron transport chain and ATP synthesis, as determined for mesocarp tissue of fruits of high- versus low-yielding oil palm plants.
  • FIG. 12 provides a heat map indicating relative levels of various metabolites of glyco lytic pathways, as determined for mesocarp tissue of fruits of high- versus low- yielding oil palm plants, at weeks 12, 14, 16, 18, 20, and 22 post-pollination (increasing darkness of shading indicates increasing relative level of metabolite).
  • FIG. 13 is a graph of relative concentration versus time (weeks 12 to 22) post- pollination for various metabolites of glycolytic pathways, as determined for mesocarp tissue of fruits of high- versus low-yielding oil palm plants.
  • the application is drawn to methods for obtaining high-yielding oil palm plants, methods for predicting oil yield of test oil palm plants, and kits for obtaining high- yielding oil palm plants.
  • the level of a metabolite in mesocarp tissue of a fruit of an oil palm plant can be used for obtaining a high-yielding oil palm plant and for predicting oil yield of a test oil palm plant.
  • Metabolites useful in this regard include 2-aminobutyric acid, 2-hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine, cyclohexylamine, cytidine, gamma-aminobutyric acid, glyoxylic acid, guanosine, homo-L-arginine, glutamine, indole-3-acetaldehyde, lysine, myo-inositol-phosphate, myo-inositol-2-phosphate, nicotinamide, ⁇ - ⁇ -methylarginine, octylamine, ophthalmic acid, serine, spermidine, triethanolamine, uracil, uridine, and beta-alanine.
  • Additional metabolites useful in this regard include l-methyladenosine, 4-aminophenylsulfone, 4-guanidinobutyric acid, 6,8- thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N-acetylhistidine, N- methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tartaric acid, tetrahydrouridine, thiamine, and tryptophan.
  • the application provides methods for obtaining high-yielding oil palm plants comprising determining the level of one of the above-noted metabolites in mesocarp tissue of a fruit of a parental oil palm plant 11 to 23 weeks after pollination thereof, determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 23 weeks after pollination thereof, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is, depending on the particular metabolite, at least 15% higher, or at least 5% lower, than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm
  • the application provides methods for predicting oil yield of test oil palm plants comprising determining the level of one of the above-noted metabolites in mesocarp tissue of a fruit of a test oil palm plant 11 to 23 weeks after pollination thereof, determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 23 weeks after pollination thereof, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant is, depending on the particular metabolite, at least 15% higher, or at least 5% lower, than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and predicting the oil yield of the test oil palm plants based on the difference.
  • kits for obtaining a high-yielding oil palm plant or predicting oil yield of a test oil palm plant comprising a reagent for quantitative detection of one of the above-noted metabolites, an extract of a mesocarp tissue of a fruit of a reference oil palm plant 1 1 to 23 weeks after pollination thereof, and instructions indicating use of the reagent for determining whether there is a difference between the level of the metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 11 to 23 weeks after pollination thereof, and the level of the metabolite in the extract of the mesocarp tissue of the fruit of the reference oil palm plant, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental or test oil palm plant is, depending on the metabolite, at least 15% higher or at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant
  • parental oil palm plant means an oil palm plant from which progeny have been generated, are generated, or will be generated during the course of carrying out methods for obtaining a high-yielding oil palm plant as disclosed herein or using kits for obtaining a high-yielding oil palm plant as disclosed herein.
  • test oil palm plant means an oil palm plant which has been subjected, is subjected, or will be subjected to a step of determining the level of a metabolite in mesocarp tissue of a fruit thereof during the course of carrying out methods for predicting oil yield of the plant as disclosed herein.
  • reference oil palm plant means an oil palm plant used as a basis for comparison in determining oil palm yield traits.
  • the reference oil palm plant can be, for example, an oil palm plant that produces high, average, or low amounts of palm oil, depending on the context of the particular application.
  • the reference oil palm plant can be an oil palm plant that produces 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 tonnes of palm oil per hectare per year.
  • high-yielding “low-yielding,” and “oil yield,” as used herein with respect to the methods and kits disclosed herein, refer to yields of palm oil in mesocarp tissue of fruits of oil palm plants.
  • a method for obtaining a high-yielding oil palm plant comprises: (i) determining the level of a metabolite in mesocarp tissue of a fruit of a parental oil palm plant 1 1 to 23 weeks after pollination thereof, (ii) determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 1 1 to 23 weeks after pollination thereof, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant; and (iii) selecting progeny of the parental oil palm plant based on the difference to obtain the high-yielding oil palm plant.
  • the metabolite is selected from the group consisting of 2-aminobutyric acid, 2-hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine, cyclohexylamine, cytidine, gamma-aminobutyric acid, glyoxylic acid, guanosine, homo-L-arginine, glutamine, indole-3-acetaldehyde, lysine, myo-inositol-phosphate, myo-inositol-2- phosphate, nicotinamide, ⁇ - ⁇ -methylarginine, octylamine, ophthalmic acid, serine, spermidine, triethanolamine, uracil, uridine, and beta-alanine.
  • metabolites can be grouped according to metabolic pathways with which they are primarily associated.
  • asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, homo-L-arginine, glutamine, lysine, co-N- methylarginine, serine, and beta-alanine are primarily associated with amino acid metabolic pathways.
  • adenine, cytidine, guanosine, uracil, and uridine are primarily associated with nucleoside metabolic pathways.
  • cadaverine and spermidine are primarily associated with polyamine metabolic pathways.
  • the metabolite is a metabolite that is primarily associated with amino acid metabolic pathways selected from the group consisting of asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, homo-L- arginine, glutamine, lysine, ⁇ - ⁇ -methylarginine, serine, and beta-alanine.
  • the metabolite is a metabolite that is primarily associated with nucleoside metabolic pathways selected from the group consisting of adenine, cytidine, guanosine, uracil, and uridine.
  • the metabolite is a metabolite that is primarily associated with polyamine metabolic pathways selected from the group consisting of cadaverine and spermidine.
  • metabolites can also be grouped according to metabolic pathways with which they are not primarily or even proximately associated. Strikingly, although each of these metabolites, i.e. 2-aminobutyric acid, 2-hydroxyva eric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine,
  • the level of one of the above-noted metabolites in mesocarp tissue of a fruit of a parental oil palm plant may be determined by any of numerous techniques that are known in the art, as described for example throughout 57 PLANT METABOLOMICS (K. Saito, R.A. Dixon & L. Willmitzer eds., 2006) and references cited therein.
  • the level of the metabolite can be determined in a preparation of metabolites from mesocarp tissue, e.g. a crude preparation, a minimally purified preparation, or a highly purified preparation of mesocarp metabolites.
  • the preparation may be obtained from an extract of mesocarp tissue, e.g.
  • lipid phase extract based on lyophilization followed by isopropanol extraction and recovery of the lipid phase in chloroform, or an aqueous phase extract based on mechanical homogenization followed by recovery of the aqueous phase in water.
  • the preparation may include total metabolites or a subset of total metabolites, e.g. metabolites soluble in polar solvents, metabolites soluble in nonpolar solvents, metabolites amenable to gas chromatographic separation, metabolites amenable to liquid chromatographic separation, or metabolites amenable to capillary electrophoretic separation, among others.
  • the mesocarp tissue of the fruit of the parental oil palm plant may be obtained and tested at a particular developmental stage of the fruit from which it is derived, e.g. 1 1 to 23 weeks after pollination thereof, i.e. after the pollination event from which the fruit originated.
  • the mesocarp tissue may be obtained and tested 1 1-21 weeks, 11-19 weeks, 11-17 weeks, 1 1-15 weeks, 11-13 weeks, 13-23 weeks, 13-21 weeks, 13-19 weeks, 13-17 weeks, 13-15 weeks, 15-23 weeks, 15-21 weeks, 15-19 weeks, 15-17 weeks, 17-23 weeks, 17-21 weeks, 17-19 weeks, 19-23 weeks, 19-21 weeks, 21-23 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, or 23 weeks after pollination thereof.
  • the step of determining the level of the metabolite in mesocarp tissue of a fruit of a parental oil palm plant may be carried out by an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, a biochemical assay, a chemical assay, a staining assay, and a chromatographic assay.
  • the step of determining the level of the metabolite in mesocarp tissue of a fruit of a parental oil palm plant may be carried out by determining the mean level of the metabolite in mesocarp tissue of at least one fruit of at least one parental oil palm plant.
  • the mean level of the metabolite may be determined, for example, by determining the levels of the metabolite in mesocarp tissue of each of a plurality of fruits of at least one parental oil palm plant, summing the levels, and then dividing the sum by the total number of fruits tested to obtain the mean level of the metabolite.
  • the mean level of the metabolite may be determined by determining the levels of the metabolite in mesocarp tissue of at least one fruit from each of a plurality of parental oil palm plants, summing the levels, and then dividing the sum by the total number of fruits tested to obtain the mean level of the metabolite.
  • the mean level of the metabolite may be determined by determining the levels of the metabolite in mesocarp tissue of a plurality of fruits from each of a plurality of parental oil palm plants, summing the levels, and then dividing the sum by the total number of fruits tested to obtain the mean level of the metabolite.
  • the mean level of the metabolite may be determined by combining mesocarp tissue from at least one fruit of at least one parental oil palm plant and then determining the level of the metabolite in the combined mesocarp tissue to obtain the mean level.
  • the mean level of the metabolite may be determined by various combinations of the above-noted approaches.
  • levels are determined for more than one of the metabolites.
  • levels are determined for a combination of two to thirty of the metabolites, i.e. two to thirty of the metabolites selected from the group consisting of 2-aminobutyric acid, 2-hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine, cyclohexylamine, cytidine, gamma-aminobutyric acid, glyoxylic acid, guanos ne, homo-L-arginine, glutamine, indole-3-acetaldehyde, lysine, myo-inositol -phosphate, myo-inositol-2- phosphate, nicotinamide, co-N-methylarginine, octylamine, ophthalmic acid, serine, sper
  • levels are determined for combinations of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of the metabolites.
  • levels are determined for 2-aminobutyric acid and one of the following: 2-hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine, cyclohexylamine, cytidine, gamma- aminobutyric acid, glyoxylic acid, guanosine, homo-L-arginine, glutamine, indole-3- acetaldehyde, lysine, myo-inositol-phosphate, myo-inositol-2-phosphate, nicotinamide, ⁇ - ⁇ -methylarginine,
  • determining the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is necessarily transformative of the mesocarp tissue.
  • the level cannot be determined, for example, merely based on appearance of the mesocarp tissue. Rather, determination of the level of the metabolite in the mesocarp tissue requires physical disruption and/or chemical treatment of the mesocarp tissue.
  • the level of the metabolite can be expressed, for example, in absolute quantitative terms, e.g. mass of metabolite per mass of mesocarp tissue, or in relative terms, e.g. intensity of signal of metabolite relative to intensity of signal of standard, or units of area under the curve ("AUC")-
  • the step of determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant may be carried out by determining the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, for example by an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, a biochemical assay, a chemical assay, a staining assay, and a chromatographic assay, as described above, and then checking for a difference therebetween.
  • the mesocarp tissue of the fruit of the reference oil palm plant may be obtained and tested at a particular developmental stage of the fruit from which it is derived, e.g. 11 to 23 weeks after pollination thereof, or 1 1-21 weeks, 11-19 weeks, 11-17 weeks, 1 1-15 weeks, 1 1-13 weeks, 13-23 weeks, 13-21 weeks, 13-19 weeks, 13-17 weeks, 13-15 weeks, 15-23 weeks, 15-21 weeks, 15-19 weeks, 15-17 weeks, 17-23 weeks, 17-21 weeks, 17-19 weeks, 19-23 weeks, 19-21 weeks, 21-23 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, or 23 weeks after pollination thereof.
  • the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant the level of the metabolite in the mesocarp tissue of the fruit of the reference
  • the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant may be determined before, during, or after determination of the level of the metabolite in mesocarp tissue of the fruit of the parental oil palm plant.
  • the step of determining whether there is a difference is considered to reveal a biologically and/or statistically significant difference based, for example, on the difference being that the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, i.e. the ratio of the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant to the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant is 1.15:1 or higher, and wherein the p value for the comparison based on a T- test is less than 0.1.
  • such a comparison is considered to reveal a biologically and/or statistically significant difference based on the difference being that the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is, for example, at least 25% higher, at least 50% higher, or at least 100% higher, than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, wherein the p value for the comparison based on a T-test is, for example, less than 0.05, less than 0.025, or less than 0.01.
  • the difference relates to a mean level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and/or the reference oil palm plant.
  • the difference is that the level of the metabolite in the mesocarp tissue of a fruit of the parental oil palm plants is higher than the mean level of the metabolite in the mesocarp tissue of at least one fruit of at least one reference oil palm plant.
  • the difference is that the mean level of the metabolite in the mesocarp tissue of at least one fruit of at least one parental oil palm plant is higher than the level of the metabolite in the mesocarp tissue of a fruit of the reference oil palm plant.
  • the difference is that the mean level of the metabolite in the mesocarp tissue of at least one fruit of at least one parental oil palm plant is higher than the mean level of the metabolite in the mesocarp tissue of at least one fruit of at least one reference oil palm plant.
  • the difference is that the mean level of the metabolite in the mesocarp tissue of a plurality of the fruits of a plurality of the parental oil palm plants is higher than the mean level of the metabolite in the mesocarp tissue of a plurality of the fruits of a plurality of the reference oil palm plants.
  • the difference may relate to various combinations of the above-noted differences.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the 2- aminobutyric acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 47% to 110% higher than the level of the 2- aminobutyric acid in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the 2- hydroxyvaleric acid in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 13 weeks after pollination thereof is 56% to 130% higher than the level of the 2- hydroxyvaleric acid in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 13 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the adenine in the mesocarp tissue of the fruit of the parental oil palm plant 1 1 to 21 weeks after pollination thereof is 110% to 240% higher than the level of the adenine in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the asymmetric dimethyl arginine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 39% to 92% higher than the level of the asymmetric dimethyl arginine in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the alanine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 67% to 150% higher than the level of the alanine in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the alanyl- alanine in the mesocarp tissue of the fruit of the parental oil palm plant 17 to 19 weeks after pollination thereof is 35% to 85% higher than the level of the alanyl-alanine in the mesocarp tissue of the fruit of the reference oil palm plant 17 to 19 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the arginine in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 15 weeks after pollination thereof is 67% to 300% higher than the level of the arginine in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the asparagine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 17 weeks after pollination thereof is 89% to 240% higher than the level of the asparagine in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 17 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the cadaverine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 21 weeks after pollination thereof is 30% to 75% higher than the level of the cadaverine in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the cyclohexylamine in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 21 weeks after pollination thereof is 35% to 79% higher than the level of the
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the cytidine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 21 weeks after pollination thereof is 59% to 170% higher than the level of the cytidine in the mesocarp tissue of the fruit of the reference oil palm plant 1 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the gamma- aminobutyric acid in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 21 weeks after pollination thereof is 39% to 120% higher than the level of the gamma- aminobutyric acid in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the glyoxylic acid in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 15 weeks after pollination thereof is 41% to 82% higher than the level of the glyoxylic acid in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the guanosine in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 21 weeks after pollination thereof is 43% to 180% higher than the level of the guanosine in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the homo-L- arginine in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 15 weeks after pollination thereof is 120% to 300% higher than the level of the homo-L-arginine in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the glutamine in the mesocarp tissue of the fruit of the parental oil palm plant 1 1 to 15 weeks after pollination thereof is 150% to 670% higher than the level of the glutamine in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the indole-3- acetaldehyde in the mesocarp tissue of the fruit of the parental oil palm plant 19 to 21 weeks after pollination thereof is 22% to 61% higher than the level of the indole-3- acetaldehyde in the mesocarp tissue of the fruit of the reference oil palm plant 19 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the lysine in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 15 weeks after pollination thereof is 41% to 110% higher than the level of the lysine in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the myo- inositol-phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 15 weeks after pollination thereof is 52% to 120% higher than the level of the myo- inositol-phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 15 weeks after pollination thereof, and further wherein the myo-inositol-phosphate is selected from the group consisting of myo-inositol-1 -phosphate and myo-inositol-3- phosphate.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the myo- inositol-2-phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 1 1 to 21 weeks after pollination thereof is 41% to 96% higher than the level of the myo- inositol-2 -phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the nicotinamide in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 21 weeks after pollination thereof is 52% to 210% higher than the level of the nicotinamide in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the co-N- methylarginine in the mesocarp tissue of the fruit of the parental oil palm plant 1 1 to 15 weeks after pollination thereof is 25% to 89% higher than the level of the co-N- methylarginine in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the octylamine in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 21 weeks after pollination thereof is 39% to 89% higher than the level of the octylamine in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the ophthalmic acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 39% to 92% higher than the level of the ophthalmic acid in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the serine in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 21 weeks after pollination thereof is 35% to 570% higher than the level of the serine in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the spermidine in the mesocarp tissue of the fruit of the parental oil palm plant 15 to 19 weeks after pollination thereof is 22% to 69% higher than the level of the spermidine in the mesocarp tissue of the fruit of the reference oil palm plant 15 to 19 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the triethanolamine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 30% to 75% higher than the level of the triethanolamine in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the uracil in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 21 weeks after pollination thereof is 47% to 190% higher than the level of the uracil in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the uridine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 21 weeks after pollination thereof is 35% to 130% higher than the level of the uridine in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the beta- alanine in the mesocarp tissue of the fruit of the parental oil palm plant 17 to 19 weeks after pollination thereof is 33% to 82% higher than the level of the beta-alanine in the mesocarp tissue of the fruit of the reference oil palm plant 17 to 19 weeks after pollination thereof.
  • the step of selecting progeny of the parental oil palm plant based on the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of the fruit of the reference oil palm plant to obtain the high-yielding oil palm plant may be carried out, for example, by choosing a parental oil palm plant for propagation based on the difference and crossing the plant with another oil palm plant, e.g. another oil palm plant also exhibiting the same or a similar difference with respect to one of the above-noted metabolites, i.e.
  • a metabolite selected from the group consisting of 2-aminobutyric acid, 2-hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine, cyclohexylamine, cytidine, gamma-aminobutyric acid, glyoxylic acid, guanosine, homo-L-arginine, glutamine, indole-3-acetaldehyde, lysine, myo-inositol-phosphate, myo-inositol-2-phosphate, nicotinamide, ⁇ - ⁇ -methylarginine, octylamine, ophthalmic acid, serine, spermidine, triethanolamine, uracil, uridine, and beta-alanine, by conventional breeding techniques to obtain progeny corresponding to the high-yielding oil palm plant.
  • fruit type is a monogenic trait in oil palm that is important with respect to breeding and commercial production of palm oil.
  • oil palms with either of two distinct fruit types are generally used in breeding and seed production through crossing in order to generate palms for commercial production of oil (also termed “commercial planting materials” or “agricultural production plants”).
  • the first fruit type is dura (genotype: sh+ sh+), which is characterized by a thick shell corresponding to 28 to 35% of the fruit by weight, with no ring of black fibres around the kernel of the fruit.
  • sh+ sh+ the mesocarp to fruit ratio varies from 50 to 60%, with extractable oil content in proportion to bunch weight of 18 to 24%.
  • the second fruit type is pisifera (genotype: sh- sh-), which is characterized by the absence of a shell, the vestiges of which are represented by a ring of fibres around a small kernel. Accordingly, for pisifera fruits, the mesocarp to fruit ratio is 90 to 100%. The mesocarp oil to bunch ratio is comparable to the dura at 16 to 28%. Pisiferas are however usually female sterile as the majority of bunches abort at an early stage of development.
  • Crossing dura and pisifera gives rise to palms with a third fruit type, the tenera (genotype: sh+ sh-).
  • Tenera fruits have thin shells of 8 to 10 % of the fruit by weight, corresponding to a thickness of 0.5 to 4 mm, around which is a characteristic ring of black fibres.
  • the ratio of mesocarp to fruit is comparatively high, in the range of 60 to 80%.
  • Commercial tenera palms generally produce more fruit bunches than duras, although mean bunch weight is lower.
  • the extractable oil to bunch ratio is in the range of 20 to 30%, the highest of the three fruit types, and thus tenera are typically used as commercial planting materials.
  • Dura palm breeding populations used in Southeast Asia include Serdang Avenue, Ulu Remis (which incorporated some Serdang Avenue material), Johor Labis, and Elmina estate, including Deli Dumpy, all of which are derived from Deli dura.
  • Pisifera breeding populations used for seed production are generally grouped as Yangambi, AVROS, Binga and URT. Other dura and pisifera populations are used in Africa and South America.
  • the parental oil palm plant is a dura palm selected from the group consisting of Deli dura, Serdang Avenue dura, Ulu Remis dura, Johor Labis dura, Elmina estate dura, and Deli Dumpy dura.
  • the parental oil palm plant is a pisifera palm selected from the group consisting of Yangambi pisifera, AVROS pisifera, Binga pisifera, and URT pisifera.
  • Oil palm breeding is primarily aimed at selecting for improved parental dura and pisifera breeding stock palms for production of superior tenera commercial planting materials. Such materials are largely in the form of seeds although the use of tissue culture for propagation of clones continues to be developed.
  • parental dura breeding populations are generated by crossing among selected dura palms. Based on the monogenic inheritance of fruit type, 100% of the resulting palms will be duras. After several years of yield recording and confirmation of bunch and fruit characteristics, duras are selected for breeding based on phenotype.
  • pisifera palms are normally female sterile and thus breeding populations thereof must be generated by crossing among selected teneras or by crossing selected teneras with selected pisiferas.
  • the tenera x tenera cross will generate 25% duras, 50% teneras and 25% pisiferas.
  • the tenera x pisifera cross will generate 50% teneras and 50% pisiferas.
  • the yield potential of pisiferas is then determined indirectly by progeny testing with the elite duras, i.e. by crossing duras and pisiferas to generate teneras, and then determining yield phenotypes of the fruits of the teneras over time. From this, pisiferas with good general combining ability are selected based on the performance of their tenera progenies. Intercrossing among selected parents is also carried out with progenies being carried forward to the next breeding cycle.
  • priority selection objectives include high oil yield per unit area in terms of high fresh fruit bunch yield and high oil to bunch ratio (thin shell, thick mesocarp), high early yield (precocity), and good oil qualities, among other traits.
  • the parental oil palm plant is a dura breeding stock plant
  • the progeny comprises an oil palm plant selected from the group consisting of a dura breeding stock plant and a tenera agricultural production plant
  • the high- yielding oil palm plant is selected from the group consisting of a dura breeding stock plant and a tenera agricultural production plant.
  • the method is carried out with the purpose of generating improved dura breeding stock, in which case the parental dura breeding stock plant is crossed with another dura breeding stock plant to obtain a high yielding oil palm plant directly among the progeny, which will also be dura breeding stock plants.
  • the method is carried out with the purpose of generating improved tenera agricultural production plants, in which case the parental dura breeding stock plant is crossed with a pisifera breeding stock plant to obtain a high yielding oil palm plant directly among the progeny, which will be tenera agricultural production plants.
  • the parental oil palm plant is a tenera breeding stock plant
  • the progeny comprises an oil palm plant selected from the group consisting of a tenera breeding stock plant, a pisifera breeding stock plant, and a tenera agricultural production plant
  • the high-yielding oil palm plant is selected from the group consisting of a tenera breeding stock plant and a tenera agricultural production plant.
  • the method may be carried out with the purpose of generating improved tenera breeding stock, in which case the parental tenera breeding stock plant is crossed with another tenera breeding stock plant, to obtain a tenera high yielding palm plant directly among the progeny, of which 25% will be dura, 50% will be tenera, and 25% will be pisifera.
  • the method is carried out with the purpose of generating improved tenera agricultural production plants, in which case the parental tenera breeding stock plant is crossed with a pisifera breeding stock plant, to yield progeny corresponding to 50% tenera and 50% pisifera. The pisifera resulting from this cross can in turn be used as pisifera breeding stock for generation of tenera agricultural production plants.
  • Progeny plants may be cultivated by conventional approaches, e.g. seedlings may be cultivated in polyethylene bags in pre-nursery and nursery settings, raised for about 12 months, and then planted as seedlings, with progeny that are known or predicted to exhibit high yields chosen for further cultivation.
  • seedlings may be cultivated in polyethylene bags in pre-nursery and nursery settings, raised for about 12 months, and then planted as seedlings, with progeny that are known or predicted to exhibit high yields chosen for further cultivation.
  • the step of selecting progeny of the parental oil palm plant may also be based on differences between levels of more than one of the metabolites in the mesocarp tissue of the fruit of the parental oil palm plant and the levels of the metabolites in mesocarp tissue of a fruit of a reference oil palm plant to obtain the high-yielding oil palm plant.
  • the step of selecting is based on differences with respect to a combination of two to thirty of the metabolites, i.e. 2-aminobutyric acid, 2-hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine,
  • cyclohexylamine cytidine, gamma-aminobutyric acid, glyoxylic acid, guanosine, homo- L-arginine, glutamine, indole-3-acetaldehyde, lysine, myo-inositol-phosphate, myoinositol ⁇ -phosphate, nicotinamide, ⁇ - ⁇ -methylarginine, octylamine, ophthalmic acid, serine, spermidine, triethanolamine, uracil, uridine, and beta-alanine.
  • the step of selecting is based on differences with respect to combinations of 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of the metabolites, e.g. each possible combination.
  • a method for obtaining palm oil from a high-yielding oil palm plant includes the steps of obtaining a high-yielding oil palm plant as explained above; and isolating palm oil from a fruit of the high- yielding oil palm plant.
  • the step of isolating palm oil may be carried out by conventional approaches, e.g. harvesting of fruit bunches followed by extraction of oil, within 24 hours, from the fresh and non-wounded fruits thereof.
  • the method comprises (i) determining the level of a metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 23 weeks after pollination thereof; (ii) determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 23 weeks after pollination thereof, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 15% higher than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant; and (iii) predicting the oil yield of the test oil palm plant based on the difference.
  • a metabolite selected from the group consisting of 2- aminobutyric acid, 2-hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine, cyclohexylamine, cytidine, gamma-aminobutyric acid, glyoxylic acid, guanosine, homo-L-arginine, glutamine, indole-3-acetaldehyde, lysine, myo-inositol-phosphate, myo-inositol-2-phosphate, nicotinamide, ⁇ - ⁇ -methyIarginine, octylamine, ophthalmic acid, serine, spermidine, triethanolamine, uracil, uridine, and beta-alan
  • the step of determining the level of a metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 23 weeks after pollination thereof may also be carried out similarly as described above, e.g. by an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, a biochemical assay, a chemical assay, a staining assay, and a chromatographic assay, and/or as a mean level, and/or with respect to more than one of the metabolites, except that the level of the metabolite in the mesocarp tissue of the fruit is determined with respect to a fruit of a test oil palm plant rather than a parental oil palm plant.
  • an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass
  • the step of determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant may also be carried out as described above, based for example on the difference being that the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 15% higher than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and wherein the p value for the comparison based on a T-test is less than 0.1.
  • the difference may be based on any of the specific differences noted above with respect to each specific metabolite, e.g. in some embodiments the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the 2-aminobutyric acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 47% to 110% higher than the level of the 2-aminobutyric acid in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after pollination thereof.
  • differences may be determined with respect to levels of more than one of the metabolites.
  • the predicting step may be carried out, for example, based on the amount of the difference in the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of the fruit of the reference oil palm plant, and/or based on correlations between levels of expression of the metabolite and yield of palm oil.
  • the predicting step also may be carried out, for example, based on differences with respect to the levels of more than one of the metabolites.
  • kits for obtaining a high-yielding oil palm plant or predicting oil yield of a test oil palm plant comprises: (i) a reagent for quantitative detection of a metabolite; (ii) an extract of a mesocarp tissue of a fruit of a reference oil palm plant 1 1 to 23 weeks after pollination thereof; and (iii) instructions indicating use of the reagent for determining whether there is a difference between the level of the metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 1 1 to 23 weeks after pollination thereof, and the level of the metabolite in the extract of the mesocarp tissue of the fruit of the reference oil palm plant, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental or test oil palm plant is at least 15% higher than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm
  • the reagent for quantitative detection of a metabolite may be any of numerous reagents useful for specific and quantitative detection of a given metabolite that are known in the art.
  • the extract of the mesocarp tissue may be any of numerous extract forms that are known in the art, e.g. a lipid phase extract based on lyophilization followed by isopropanol extraction and recovery of the lipid phase in chloroform, or an aqueous phase extract based on mechanical homogenization followed by recovery of the aqueous phase in water, as discussed above.
  • a method for obtaining a high-yielding oil palm plant comprises: (i) determining the level of a metabolite in mesocarp tissue of a fruit of a parental oil palm plant 11 to 23 weeks after pollination thereof, (ii) determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 1 1 to 23 weeks after pollination thereof, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant; and (iii) selecting progeny of the parental oil palm plant based on the difference to obtain the high-yielding oil palm plant.
  • the metabolite is a metabolite selected from the group consisting of 1 -methyladenosine, 4-aminophenylsulfone, 4-guanidinobutyric acid, 6,8-thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N-acetylhistidine, N- methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tartaric acid, tetrahydrouridine, thiamine, and tryptophan.
  • metabolites can also be grouped according to metabolic pathways with which they are primarily associated. For example, N6-acetyllysine, N6-methyladenine, N-acetylhistidine, N-methylproline, symmetric dimethyl arginine, and tryptophan are primarily associated with amino acid metabolic pathways. In comparison, 1- methyladenosine, cytosine, and tetrahydrouridine are primarily associated with nucleoside metabolic pathways.
  • the metabolite is a metabolite that is primarily associated with amino acid metabolic pathways selected from the group consisting of N6- acetyllysine, N6-methyladenine, N-acetylhistidine, N-methylproline, symmetric dimethyl arginine, and tryptophan.
  • the metabolite is a metabolite that is primarily associated with nucleoside metabolic pathways selected from the group consisting of 1 -methyladenosine, cytosine, and tetrahydrouridine.
  • metabolites can also be grouped according to metabolic pathways with which they are not primarily or even proximately associated. Strikingly, although each of these metabolites, i.e. 1 -methyladenosine, 4-aminophenylsulfone, 4-guanidinobutyric acid, 6,8-thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N-acetylhistidine, N- methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tartaric acid, tetrahydrouridine, thiamine, and tryptophan, can be used for
  • the level of the metabolite i.e. a metabolite selected from the group consisting of 1 -methyladenosine, 4-aminophenylsulfone, 4- guanidinobutyric acid, 6,8-thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N- acetylhistidine, N-methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tartaric acid, tetrahydrouridine, thiamine, and tryptophan, in mesocarp tissue of a fruit of a parental oil
  • the level may be determined in a preparation of metabolites from mesocarp tissue, e.g. a crude preparation, a minimally purified preparation, or a highly purified preparation of mesocarp metabolites.
  • the preparation may include total metabolites or a subset of total metabolites.
  • the mesocarp tissue of the fruit of the parental oil palm plant may be obtained and tested at a particular developmental stage of the fruit from which it is derived, e.g.
  • 1 1 to 23 weeks after pollination thereof for example 11-21 weeks, 1 1-19 weeks, 1 1-17 weeks, 11-15 weeks, 11-13 weeks, 13-23 weeks, 13-21 weeks, 13-19 weeks, 13-17 weeks, 13-15 weeks, 15-23 weeks, 15-21 weeks, 15-19 weeks, 15-17 weeks, 17-23 weeks, 17-21 weeks, 17-19 weeks, 19-23 weeks, 19-21 weeks, 21-23 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, or 23 weeks after pollination thereof.
  • the step of determining the level of the metabolite in mesocarp tissue of a fruit of a parental oil palm plant may be carried out by an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, a biochemical assay, a chemical assay, a staining assay, and a
  • the step of determining the level of the metabolite in mesocarp tissue of a fruit of a parental oil palm plant may be carried out by determining the mean level of the metabolite in mesocarp tissue of at least one fruit of at least one parental oil palm plant.
  • the mean level of the metabolite may be determined, for example, by determining the levels of the metabolite in mesocarp tissue of each of a plurality of fruits of at least one parental oil palm plant, summing the levels, and then dividing the sum by the total number of fruits tested to obtain the mean level of the metabolite.
  • the mean level of the metabolite may be determined by determining the levels of the metabolite in mesocarp tissue of at least one fruit from each of a plurality of parental oil palm plants, summing the levels, and then dividing the sum by the total number of fruits tested to obtain the mean level of the metabolite.
  • the mean level of the metabolite may be determined by determining the levels of the metabolite in mesocarp tissue of a plurality of fruits from each of a plurality of parental oil palm plants, summing the levels, and then dividing the sum by the total number of fruits tested to obtain the mean level of the metabolite.
  • the mean level of the metabolite may be determined by combining mesocarp tissue from at least one fruit of at least one parental oil palm plant and then determining the level of the metabolite in the combined mesocarp tissue to obtain the mean level.
  • the mean level of the metabolite may be determined by combinations of the above-noted approaches.
  • levels are determined for more than one of the metabolites.
  • levels are determined for a combination of two to twenty-eight of the metabolites, i.e. two to twenty-eight of the metabolites selected from the group consisting of 1 -methyladenosine, 4- aminophenylsulfone, 4-guanidinobutyric acid, 6,8-thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6- methyladenine, N-acetylhistidine, N-methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tartaric acid, te
  • determining the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is necessarily transformative of the mesocarp tissue, and the level of the metabolite can be expressed in absolute quantitative terms or in relative terms.
  • the step of determining whether there is a difference between the level of the metabolite i.e. a metabolite selected from the group consisting of 1-methyladenosine, 4-aminophenylsulfone, 4-guanidinobutyric acid, 6,8- thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N-acetylhistidine, N- methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tartaric acid, tetrahydrouridine, thiamine, and tryptophan, in the mesocar
  • a metabolite
  • the step of determining whether there is a difference may be carried out by comparing the respective levels of the metabolite, for example by an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, a biochemical assay, a chemical assay, a staining assay, and a
  • the mesocarp tissue of the fruit of the parental oil palm plant may be obtained and tested at a particular developmental stage of the fruit from which it is derived, e.g.
  • 1 1 to 23 weeks after pollination thereof and more particularly 1 1-21 weeks, 1 1-19 weeks, 1 1 -17 weeks, 11-15 weeks, 11-13 weeks, 13-23 weeks, 13-21 weeks, 13-19 weeks, 13-17 weeks, 13-15 weeks, 15-23 weeks, 15-21 weeks, 15-19 weeks, 15-17 weeks, 17-23 weeks, 17-21 weeks, 17-19 weeks, 19-23 weeks, 19-21 weeks, 21-23 weeks, 1 1 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, or 23 weeks after pollination thereof.
  • the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant may be determined as a mean level.
  • the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant may be determined before, during, or after determination of the level of the metabolite in mesocarp tissue of the fruit of the parental oil palm plant.
  • the step of determining whether there is a difference is considered to reveal a biologically and/or statistically significant difference based, for example, on the difference being that the level of the metabolite, i.e. a metabolite selected from the group consisting of 1 -methyladenosine, 4-aminophenylsulfone, 4- guanidinobutyric acid, 6,8-thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, giucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N- acetylhistidine, N-methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tartaric acid,
  • the ratio of the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant to the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant is 0.95: 1 or lower, and wherein the p value for the comparison based on a T-test is less than 0.1.
  • such a comparison is considered to reveal a biologically and/or statistically significant difference based on the difference being that the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is, for example, at least 10% lower, at least 25% lower, or at least 50% lower, than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, wherein the p value for the comparison based on a T-test is, for example, less than 0.05, less than 0.025, or less than 0.01.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the 1- methyladenosine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 17 weeks after pollination thereof is 28% to 44% lower than the level of the 1- methyladenosine in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 17 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the 4- aminophenylsulfone in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 9.9% to 34% lower than the level of the 4- aminophenylsulfone in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the 4- guanidinobutyric acid in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 17 weeks after pollination thereof is 22% to 34% lower than the level of the 4- guanidinobutyric acid in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 17 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the 6,8- thioctic acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 21 weeks after pollination thereof is 26% to 44% lower than the level of the 6,8-thioctic acid in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the ascorbic acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 21 weeks after pollination thereof is 22% to 57% lower than the level of the ascorbic acid in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the chelidonic acid in the mesocarp tissue of the fruit of the parental oil palm plant 1 1 to 21 weeks after pollination thereof is 17% to 44% lower than the level of the chelidonic acid in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the cytosine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 17 weeks after pollination thereof is 21% to 40% lower than the level of the cytosine in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 17 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the etidronic acid in the mesocarp tissue of the fruit of the parental oil palm plant 15 to 17 weeks after pollination thereof is 17% to 38% lower than the level of the etidronic acid in the mesocarp tissue of the fruit of the reference oil palm plant 15 to 17 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the glucaric acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 25% to 42% lower than the level of the glucaric acid in the mesocarp tissue of the fruit of the reference oil palm plant 1 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the glutathione disulfide in the mesocarp tissue of the fruit of the parental oil palm plant 1 to 15 weeks after pollination thereof is 23% to 41% lower than the level of the glutathione disulfide in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the imidazoIe-4-acetic acid in the mesocarp tissue of the fruit of the parental oil palm plant 19 to 21 weeks after pollination thereof is 47% to 57% lower than the level of the imidazole-4-acetic acid in the mesocarp tissue of the fruit of the reference oil palm plant 19 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the imidazolelactic acid in the mesocarp tissue of the fruit of the parental oil palm plant 17 to 21 weeks after pollination thereof is 30% to 53% lower than the level of the
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the methyl sulfate in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 21 weeks after pollination thereof is 32% to 49% lower than the level of the methyl sulfate in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the metronidazole in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 17 weeks after pollination thereof is 23% to 36% lower than the level of the metronidazole in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 17 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the mucic acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 17 weeks after pollination thereof is 25% to 41% lower than the level of the mucic acid in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 17 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the N6- acetyllysine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 31% to 46% lower than the level of the N6-acetyllysine in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the N6- methyladenine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 37% to 50% lower than the level of the N6- methyladenine in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the N- acetylhistidine in the mesocarp tissue of the fruit of the parental oil palm plant 19 to 21 weeks after pollination thereof is 26% to 43 % lower than the level of the N- acetylhistidine in the mesocarp tissue of the fruit of the reference oil palm plant 19 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the N- methylproline in the mesocarp tissue of the fruit of the parental oil palm plant 19 to 21 weeks after pollination thereof is 27% to 44% lower than the level of the N-methylproline in the mesocarp tissue of the fruit of the reference oil palm plant 19 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the pantothenic acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 18% to 38% lower than the level of the pantothenic acid in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the phosphorylcholine in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 28% to 44% lower than the level of the
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the quinic acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 21 weeks after pollination thereof is 18% to 45% lower than the level of the quinic acid in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the symmetric dimethyl arginine in the mesocarp tissue of the fruit of the parental oil palm plant 19 to 21 weeks after pollination thereof is 27% to 44% lower than the level of the symmetric dimethyl arginine in the mesocarp tissue of the fruit of the reference oil palm plant 19 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the stachydrine in the mesocarp tissue of the fruit of the parental oil palm plant 17 to 19 weeks after pollination thereof is 58% to 64% lower than the level of the stachydrine in the mesocarp tissue of the fruit of the reference oil palm plant 17 to 19 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the tartaric acid in the mesocarp tissue of the fruit of the parental oil palm plant 19 to 21 weeks after pollination thereof is 42% to 53% lower than the level of the tartaric acid in the mesocarp tissue of the fruit of the reference oil palm plant 19 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the tetrahydrouridine in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 15 weeks after pollination thereof is 17% to 34% lower than the level of the
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the thiamine in the mesocarp tissue of the fruit of the parental oil palm plant 17 to 19 weeks after pollination thereof is 31% to 46% lower than the level of the thiamine in the mesocarp tissue of the fruit of the reference oil palm plant 17 to 19 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the tryptophan in the mesocarp tissue of the fruit of the parental oil palm plant 15 to 17 weeks after pollination thereof is 68% to 72% lower than the level of the tryptophan in the mesocarp tissue of the fruit of the reference oil palm plant 15 to 17 weeks after pollination thereof.
  • the step of selecting progeny may be carried out by choosing a parental oil palm plant for propagation based on the difference and crossing the plant with another oil palm plant, e.g. another oil palm plant also exhibiting the same or a similar difference with respect to one of the metabolites, by conventional breeding techniques to obtain progeny corresponding to the high-yielding oil palm plant.
  • the parental oil palm plant is a dura breeding stock plant
  • the progeny comprises an oil palm plant selected from the group consisting of a dura breeding stock plant and a tenera agricultural production plant
  • the high-yielding oil palm plant is selected from the group consisting of a dura breeding stock plant and a tenera agricultural production plant.
  • the parental oil palm plant is a tenera
  • the progeny comprises an oil palm plant selected from the group consisting of a tenera breeding stock plant, a pisifera breeding stock plant, and a tenera agricultural production plant
  • the high-yielding oil palm plant is selected from the group consisting of a tenera breeding stock plant and a tenera agricultural production plant.
  • progeny plants may be cultivated by conventional approaches and then planted as seedlings, with progeny that are known or predicted to exhibit high yields chosen for further cultivation.
  • the step of selecting progeny of the parental oil palm plant may also be based on differences between levels of more than one of the metabolites in the mesocarp tissue of the fruit of the parental oil palm plant and the levels of the metabolites in mesocarp tissue of a fruit of a reference oil palm plant to obtain the high- yielding oil palm plant.
  • the step of selecting is based on differences with respect to a combination of two to twenty-eight of the metabolites, i.e.
  • adenosine 4-aminophenylsulfone, 4-guanidinobutyric acid, 6,8- thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N-acetylhistidine, N- methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tartaric acid, tetrahydrouridine, thiamine, and tryptophan.
  • the step of selecting is based on differences with respect to combinations of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 of the metabolites, e.g. each possible combination.
  • a method for obtaining palm oil from a high-yielding oil palm plant includes the steps of obtaining a high-yielding oil palm plant as explained above; and isolating palm oil from a fruit of the high-yielding oil palm plant.
  • the step of isolating palm oil may be carried out by conventional approaches, e.g. harvesting of fruit bunches followed by extraction of oil, within 24 hours, from the fresh and non-wounded fruits thereof.
  • the method comprises (i) determining the level of a metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 23 weeks after pollination thereof; (ii) determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 1 1 to 23 weeks after pollination thereof, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant; and (iii) predicting the oil yield of the test oil palm plant based on the difference.
  • metabolites noted above as being useful in the other method for obtaining a high- yielding oil palm plant i.e. a metabolite selected from the group consisting of 1- methyladenosine, 4-aminophenylsulfone, 4-guanidinobutyric acid, 6,8-thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N-acetylhistidine, N-methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tartaric acid, tetrahydrouridine, thiamine, and tryptophan, are also useful in the method for predicting oil
  • the step of determining the level of a metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 23 weeks after pollination thereof may also be carried out similarly as described above, e.g. by an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, a biochemical assay, a chemical assay, a staining assay, and a chromatographic assay, and/or as a mean level, and/or with respect to more than one of the metabolites, except that the level of the metabolite in the mesocarp tissue of the fruit is determined with respect to a fruit of a test oil palm plant rather than a parental oil palm plant.
  • an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass
  • the step of determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant may also be carried out as described above, based for example on the difference being that the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and wherein the p value for the comparison based on a T-test is less than 0.1.
  • the difference may be based on any of the specific differences noted above with respect to each specific metabolite, e.g. in some embodiments the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the 1-methyladenosine in the mesocarp tissue of the fruit of the test oil palm plant 13 to 17 weeks after pollination thereof is 28% to 44% lower than the level of the 1-methyladenosine in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 17 weeks after pollination thereof.
  • differences may be determined with respect to levels of more than one of the metabolites.
  • the predicting step may be carried out, for example, based on the amount of the difference in the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of the fruit of the reference oil palm plant, and/or based on correlations between levels of expression of the metabolite and yield.
  • the predicting step also may be carried out, for example, based on differences with respect to the levels of more than one of the metabolites.
  • kits for obtaining a high-yielding oil palm plant or predicting oil yield of a test oil palm plant comprises: (i) a reagent for quantitative detection of a metabolite; (ii) an extract of a mesocarp tissue of a fruit of a reference oil palm plant 11 to 23 weeks after pollination thereof; and (iii) instructions indicating use of the reagent for determining whether there is a difference between the level of the metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 11 to 23 weeks after pollination thereof, and the level of the metabolite in the extract of the mesocarp tissue of the fruit of the reference oil palm plant, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental or test oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the reagent for quantitative detection of a metabolite may be any of numerous reagents useful for specific and quantitative detection of a given metabolite that are known in the art
  • the extract of the mesocarp tissue may be any of numerous extract forms that are known in the art.
  • an additional method for obtaining a high- yielding oil palm plant comprises: (i) determining the level of a metabolite in mesocarp tissue of a fruit of a parental oil palm plant 1 1 to 23 weeks after pollination thereof, (ii) determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 23 weeks after pollination thereof, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant; and (iii) selecting progeny of the parental oil palm plant based on the difference to obtain the high-yielding oil palm plant.
  • the metabolite is a metabolite selected from the group consisting of adenosine triphosphate and uridine triphosphate.
  • Adenosine triphosphate and uridine triphosphate are both ribonucleotides that are substrates for synthesis of RNA during transcription and that serve as sources of energy and as an activators of substrates in metabolic reactions. Strikingly, although fatty acid
  • biosynthesis and triacylglycerol biosynthesis require sources of energy and activators of substrates, it would not have been apparent whether levels of adenosine triphosphate and uridine triphosphate in mesocarp tissue of a fruit of a parental oil palm plant exhibiting increased production of fatty acids and triacylglycerols would have been higher, the same, or lower, than levels in mesocarp tissue of a fruit of a reference oil palm plant, e.g.
  • the level of one of the metabolites i.e. a metabolite selected from the group consisting of adenosine triphosphate and uridine triphosphate, in mesocarp tissue of a fruit of a parental oil palm plant may also be determined as described above.
  • the level may be determined in a preparation of metabolites from mesocarp tissue, e.g. a crude preparation, a minimally purified preparation, or a highly purified preparation of mesocarp metabolites.
  • the preparation may include total metabolites or a subset of total metabolites.
  • the mesocarp tissue of the fruit of the parental oil palm plant may be obtained and tested at a particular developmental stage of the fruit from which it is derived, e.g. 1 1 to 23 weeks after pollination thereof, for example 11-21 weeks, 11-19 weeks, 1 1-17 weeks, 1 1-15 weeks, 1 1-13 weeks, 13-23 weeks, 13-21 weeks, 13-19 weeks, 13-17 weeks, 13-15 weeks, 15-23 weeks, 15-21 weeks, 15-19 weeks, 15-17 weeks, 17-23 weeks, 17-21 weeks, 17-19 weeks, 19-23 weeks, 19-21 weeks, 21-23 weeks, 1 1 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, or 23 weeks after pollination thereof.
  • the step of determining the level of the metabolite in mesocarp tissue of a fruit of a parental oil palm plant may be carried out by an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, a biochemical assay, a chemical assay, a staining assay, and a
  • the step of determining the level of the metabolite in mesocarp tissue of a fruit of a parental oil palm plant may be carried out by determining the mean level of the metabolite in mesocarp tissue of at least one fruit of at least one parental oil palm plant.
  • the mean level of the metabolite may be determined, for example, by determining the levels of the metabolite in mesocarp tissue of each of a plurality of fruits of at least one parental oil palm plant, summing the levels, and then dividing the sum by the total number of fruits tested to obtain the mean level of the metabolite.
  • the mean level of the metabolite may be determined by determining the levels of the metabolite in mesocarp tissue of at least one fruit from each of a plurality of parental oil palm plants, summing the levels, and then dividing the sum by the total number of fruits tested to obtain the mean level of the metabolite.
  • the mean level of the metabolite may be determined by determining the levels of the metabolite in mesocarp tissue of a plurality of fruits from each of a plurality of parental oil palm plants, summing the levels, and then dividing the sum by the total number of fruits tested to obtain the mean level of the metabolite.
  • the mean level of the metabolite may be determined by combining mesocarp tissue from at least one fruit of at least one parental oil palm plant and then determining the level of the metabolite in the combined mesocarp tissue to obtain the mean level.
  • the mean level of the metabolite may be determined by combinations of the above -noted approaches.
  • levels are determined for both adenosine triphosphate and uridine triphosphate.
  • determining the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is necessarily transformative of the mesocarp tissue, and the level of the metabolite can be expressed in absolute quantitative terms or in relative terms.
  • the step of determining whether there is a difference between the level of the metabolite, i.e. a metabolite selected from the group consisting of adenosine triphosphate and uridine triphosphate, in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant may also be carried out as described above.
  • the step of determining whether there is a difference may be carried out by comparing the respective levels of the metabolite, for example by an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, a biochemical assay, a chemical assay, a staining assay, and a
  • the mesocarp tissue of the fruit of the parental oil palm plant may be obtained and tested at a particular developmental stage of the fruit from which it is derived, e.g.
  • 1 1 to 23 weeks after pollination thereof and more particularly 11-21 weeks, 11-19 weeks, 11- 17 weeks, 11-15 weeks, 1 1-13 weeks, 13-23 weeks, 13-21 weeks, 13-19 weeks, 13-17 weeks, 13-15 weeks, 15-23 weeks, 15-21 weeks, 15-19 weeks, 15-17 weeks, 17-23 weeks, 17-21 weeks, 17-19 weeks, 19-23 weeks, 19-21 weeks, 21-23 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, or 23 weeks after pollination thereof.
  • the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant may be determined as a mean level.
  • the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant may be determined before, during, or after determination of the level of the metabolite in mesocarp tissue of the fruit of the parental oil palm plant.
  • the step of determining whether there is a difference is considered to reveal a biologically and/or statistically significant difference based, for example, on the difference being that the level of the metabolite, i.e. a metabolite selected from the group consisting of adenosine triphosphate and uridine triphosphate, in the mesocarp tissue of the fruit of the parental oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, i.e.
  • the level of the metabolite i.e. a metabolite selected from the group consisting of adenosine triphosphate and uridine triphosphate
  • the ratio of the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant to the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant is 0.95: 1 or lower, and wherein the p value for the comparison based on a T-test is less than 0.1.
  • such a comparison is considered to reveal a biologically and/or statistically significant difference based on the difference being that the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is, for example, at least 10% lower, at least 25% lower, or at least 50% lower, than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, wherein the p value for the comparison based on a T-test is, for example, less than 0.05, less than 0.025, or less than 0.01.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the adenosine triphosphate in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 21 weeks after pollination thereof is 33% to 55% lower than the level of the adenosine triphosphate in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 21 weeks after pollination thereof.
  • the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the uridine triphosphate in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 21 weeks after pollination thereof is 20% to 62% lower than the level of the uridine triphosphate in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 21 weeks after pollination thereof.
  • the step of selecting progeny of the parental oil palm plant based on the difference between the level of the metabolite, i.e. a metabolite selected from the group consisting of adenosine triphosphate and uridine triphosphate, in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of the fruit of the reference oil palm plant to obtain the high-yielding oil palm plant may be carried out as described above.
  • the step of selecting progeny may be carried out by choosing a parental oil palm plant for propagation based on the difference and crossing the plant with another oil palm plant, e.g.
  • the parental oil palm plant is a dura breeding stock plant
  • the progeny comprises an oil palm plant selected from the group consisting of a dura breeding stock plant and a tenera
  • the high-yielding oil palm plant is selected from the group consisting of a dura breeding stock plant and a tenera agricultural production plant.
  • the parental oil palm plant is a tenera breeding stock plant
  • the progeny comprises an oil palm plant selected from the group consisting of a tenera breeding stock plant, a pisifera breeding stock plant, and a tenera agricultural production plant
  • the high-yielding oil palm plant is selected from the group consisting of a tenera breeding stock plant and a tenera agricultural production plant.
  • progeny plants may be cultivated by conventional approaches and then planted as seedlings, with progeny that are known or predicted to exhibit high yields chosen for further cultivation.
  • the step of selecting progeny of the parental oil palm plant may also be based on differences between levels of more than one of the metabolites in the mesocarp tissue of the fruit of the parental oil palm plant and the levels of the metabolites in mesocarp tissue of a fruit of a reference oil palm plant to obtain the high-yielding oil palm plant.
  • the step of selecting is based on differences with respect to a combination of adenosine triphosphate and uridine triphosphate.
  • a method for obtaining palm oil from a high-yielding oil palm plant includes the steps of obtaining a high-yielding oil palm plant as explained above; and isolating palm oil from a fruit of the high-yielding oil palm plant.
  • the step of isolating palm oil may be carried out by conventional approaches, e.g. harvesting of fruit bunches followed by extraction of oil, within 24 hours, from the fresh and non-wounded fruits thereof.
  • an additional method for predicting oil yield of a test oil palm plant comprises (i) determining the level of a metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 23 weeks after pollination thereof; (ii) determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 1 1 to 23 weeks after pollination thereof, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant; and (iii) predicting the oil yield of the test oil palm plant based on the difference.
  • metabolites noted above as being useful in the additional method for obtaining a high-yielding oil palm plant i.e. a metabolite selected from the group consisting of adenosine triphosphate and uridine triphosphate, are also useful in the method for predicting oil yield of a test oil palm plant.
  • the step of determining the level of a metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 23 weeks after pollination thereof may also be carried out similarly as described above, e.g. by an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass spectrometry, a biochemical assay, a chemical assay, a staining assay, and a chromatographic assay, and/or as a mean level, and/or with respect to more than one of the metabolites, except that the level of the metabolite in the mesocarp tissue of the fruit is determined with respect to a fruit of a test oil palm plant rather than a parental oil palm plant.
  • an analytical technique selected from the group consisting of gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, capillary electrophoresis-mass
  • the step of determining whether there is a difference between the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant may also be carried out as described above, based for example on the difference being that the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and wherein the p value for the comparison based on a T-test is less than 0.1.
  • the difference may be based on any of the specific differences noted above with respect to each specific metabolite, e.g. in some embodiments the difference between the level of the metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the metabolite in mesocarp tissue of a fruit of a reference oil palm plant is that the level of the adenosine triphosphate in the mesocarp tissue of the fruit of the test oil palm plant 1 1 to 21 weeks after pollination thereof is 33% to 55% lower than the level of the adenosine triphosphate in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 21 weeks after pollination thereof.
  • differences may be determined with respect to levels of more than one of the metabolites.
  • the predicting step may be carried out, for example, based on the amount of the difference in the level of the metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the metabolite in mesocarp tissue of the fruit of the reference oil palm plant, and/or based on correlations between levels of expression of the metabolite and yield.
  • the predicting step also may be carried out, for example, based on differences with respect to the levels of more than one of the metabolites.
  • kits for obtaining a high-yielding oil palm plant or predicting oil yield of a test oil palm plant comprises: (i) a reagent for quantitative detection of a metabolite; (ii) an extract of a mesocarp tissue of a fruit of a reference oil palm plant 11 to 23 weeks after pollination thereof; and (iii) instructions indicating use of the reagent for determining whether there is a difference between the level of the metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 11 to 23 weeks after pollination thereof, and the level of the metabolite in the extract of the mesocarp tissue of the fruit of the reference oil palm plant, wherein the difference is that the level of the metabolite in the mesocarp tissue of the fruit of the parental or test oil palm plant is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant
  • metabolites described above as being useful in the additional method for obtaining a high-yielding oil palm plant i.e. a metabolite selected from the group consisting of adenosine triphosphate and uridine triphosphate, are also useful in the kit for obtaining a high-yielding oil palm plant.
  • the reagent for quantitative detection of a metabolite may be any of numerous reagents useful for specific and quantitative detection of a given metabolite that are known in the art
  • the extract of the mesocarp tissue may be any of numerous extract forms that are known in the art.
  • Objectives included identifying metabolites that are present at different levels in mesocarp tissue of fruits of high-yielding palms versus reference palms at a single time point at the onset of lipid production, for purposes of obtaining high-yielding oil palm plants and predicting palm oil yields of test oil palm plants.
  • the screening populations were derived from crosses of Serdang Avenue dura (at least 75% of Serdang Avenue dura) and AVROS pisifera (at least 75% of AVROS pisifera) to yield tenera progeny. More specifically, the high-yielding screening population was derived from a population of oil palm plants that had previously been determined to yield relatively high amounts of palm oil, specifically 10 to 12 tonnes of palm oil per hectare per year, and thus to have a high-yielding phenotype, also termed an HY phenotype.
  • the low-yielding screening population was derived from a population of oil palm plants that had previously been determined to yield relatively low amounts of palm oil, specifically 4 to 7 tonnes of palm oil per hectare per year, and thus to have a low-yielding phenotype, also termed an LY phenotype. Both populations were derived from a Carey Island oil palm plantation. The yield determinations were based on historical oil yield data for each sample.
  • LC-MS profiling was conducted on the polar (top) layer as follows.
  • LC-MS data were acquired using Accela-LTQ Orbitrap brand instrument (Thermo Fisher, Germany). Sample analysis was carried out in positive and negative ion modes of detection.
  • the mass scanning range was 100 to 2000 m/z, while capillary temperature was 300° C and sheath gas auxiliary gas flow rates were 35 and 15 arbitrary units ("arb"), respectively.
  • the sweep gas flow rate was set at 1 arb I-spray voltage at 4.5 kb.
  • the resolution was 30,000 at 1 microscan and maximum injection time at 500 ms.
  • the capillary voltage and tube lens were set at 40 V and 80 V, respectively for positive ion modes.
  • the gradient was as follows: 1% B (0 - 1.8 min), 10% B (3 min) to 40% B at 20 min and hold for 3 min, 90% B at 26-28 min and 1 % B at 29-35 min.
  • the raw data were processed and compared using Sieve version 1.2 (Thermo Fisher, Alpha Analytical, Malaysia) with the frame time and m/z width set at 1.5 min and 0.002, respectively.
  • the resulting filtrates were desiccated and then dissolved with 50 ⁇ ]_, of Milli-Q water.
  • the products were then subjected to preliminary analysis to determined peak shapes and intensities. Based on these results, the products were diluted by 50% or 20% and subjected to CE-TOFMS analysis in cation or anion modes, respectively.
  • the run buffer was Cation Buffer Solution (p/n: H3301-1001), and the rinse buffer was Cation Buffer Solution (p/n: H3301-1001).
  • Sample injection pressure was 50 mbar (10 sec).
  • the MS parameters were as follows: CE voltage : Positive, 27 kV; MS ionization : ESI Positive; MS capillary voltage : 4,000 V; MS scan range : m/z 50-1,000; Sheath liquid : HMT Sheath Liquid (p/n : H3301-1020).
  • the run buffer was Anion Buffer Solution (p/n: H3302- 1021), and the rinse buffer was Anion Buffer Solution (p/n: H3302-1022).
  • Sample injection pressure was 50 mbar (25 sec).
  • the MS parameters were as follows: CE voltage : Positive, 30 kV; MS ionization : ESI Negative; MS capillary voltage : 3,500 V; MS scan range : m/z 50- 1 ,000; Sheath liquid : HMT Sheath Liquid (p/n: H3301-1020).
  • PCA Principle component analysis
  • OPLS-DA Orthogonal Partial Least Square- Discrimination Analysis
  • PCA results of metabolome data are shown in FIG. 1.
  • PCA of the five high- yielding palms (designated H3, H5, H7, H8, and H9) and five low-yielding palms (designated L3, L4, L5, L7, and L8) batch shows clear clustering.
  • This method focuses on the variation between specified groups. Accordingly, the difference between the two groups can be comprehensively detected with the areas or relative intensities of the peaks in the chromatograms, as shown for example in FIG. 2 with respect to spermine.
  • OPLS- DA S-plot supervised analysis can be used to more clearly identify compounds of interest and to determine which compounds are most responsible for the clustering of high and low yielding samples, as shown in FIG. 3.
  • spermidine and spermine were identified among various metabolites that are present at significantly higher levels in mesocarp tissues of fruits of high-yielding oil palm plants compared to low-yielding oil palm plants.
  • spermidine was identified by comparing MS/MS fragmentation data (peak ID 255, m/z 146.16571(M+H)+, time 1.27, based on LC-MS positive mode) with the accurate mass of spermidine.
  • Spermine was also so identified (peak ID 14, m/z 203.22334 (M+H)+, time 1.278, also based on LC-MS positive mode).
  • the differences of m/z of spermidine and spermine with their accurate masses were 3.7 ppm and 1.97 ppm, respectively.
  • both spermidine, m/z 146.16571(M+H)+, and spermine, m/z 203.22334 (M+H)+ gave the product ion m/z 129.17, which corresponds to the loss of an amino group and diaminopropane, respectively.
  • MS fragmentation patterns of spermidine and spermine practically identical to those observed here have been reported previously. See, for example, Ippolito, J.E. et al., An Integrated Functional Genomics and Metabolomics Approach for Defining Poor Prognosis in Human Neuroendocrine Cancers, 102 Proc. Natl. Acad. Sci. USA 9901-9906 (2005).
  • metabolites were identified as being present at higher levels in mesocarp tissue of fruits of high-yielding oil palm plants in comparison to low-yielding oil palm plants, specifically spermidine, spermine, ornithine, glutamine, adenine, tyramine, and glycerol-3-phosphate, as indicated in TABLE 1.
  • metabolites were identified as being present at higher levels in mesocarp tissue of fruits of low- yielding oil palm plants in comparison to high-yielding oil palm plants, specifically tryptophan, 2-phosphoglyceric acid, 3-phosphoglyceric acid, gluconic acid, cytosine, uridine triphosphate ("UTP"), and adenosine triphosphate ("ATP”), as also indicated in TABLE 1.
  • UTP uridine triphosphate
  • ATP adenosine triphosphate
  • Extensive metabolite profiling was carried out essentially as described above in Example 1 in order to identify additional metabolite-based markers useful for obtaining high- yielding oil palm plants and predicting palm oil yield of test oil palm plants.
  • palms were selected from each of the high- and low-yielding screening populations as described in Example 1, i.e. a high-yielding population previously determined to yield 10 to 12 tonnes of palm oil per hectare per year, and a low-yielding population previously determined to yield 4 to 7 tonnes of palm oil per hectare per year.
  • Fruits were harvested from plants at weeks 12, 14, 16, 18, 20, and 22 post-pollination to represent fruit development before, during, and after lipid biosynthesis and ripening occurs.
  • Profiling of the two populations was carried out using CE-MS with the same extraction and acquisition parameters described in Example 1. Secondary profiling was carried out using LC-MS (LTQ-Orbitrap as in Example 1 , and LC-triple quadrupole MS as described below) as well as GC-MS in order to investigate the instrument-specificity of the results.
  • CE-MS method As described in Example 1.
  • the elution gradient was as follows: initial hold at 95% solvent A; 0 to 3 min linear gradient to 60% solvent A; 3 to 5 min 5% solvent A; 5 to 5.1 min linear gradient to 95% solvent A and hold on to 7 min. Injection volume was 3 ⁇ ⁇ .
  • the mass spectrometry was operated in both positive and negative mode with multiple reaction monitoring using ESI.
  • the capillary voltage was set at 2.9 kV, and desolvation gas was set at 800 L/hr at temperature of 350° C.
  • the collision gas flow was set at 0.15 mL/min.
  • the MRM settings in the MS/MS function with corresponding cone voltage and collision energy were optimized for each standard compound.
  • Auto dwell times were set for positive mode and negative mode, respectively.
  • Total acquisition duration for both UPLC and MS was set at 15 min. Data were acquired and processed using MassLynx V4.1 and TargetLynx, respectively.
  • Standard compounds were weighed and dissolved with 5% (v/v) acetonitrile in MilliQ water to make a stock solution with final concentration of 1 mg/mL (1000 ppm).
  • the stock solutions were diluted to 1 ppm as working stock solutions.
  • a mix standard at concentration of 1 ppm was prepared and injected into LC-MS daily to check system sensitivity and reproducibility.
  • the injection port was splitless at 280° C while column flow was maintained constant at 1 mL/min of Helium gas.
  • the MS source was set at 230° C with a scanning range of m/z 50 to m/z 650. Sample of 1 ⁇ L ⁇ was injected into a programmable injector. The total run time is 55 minutes. Data analysis:
  • FIGS. 4-13 Results are shown in FIGS. 4-13.
  • FIGS. 4 and 5 provide heat maps data and graphs of relative concentration versus time post-pollination for various metabolites of the citric acid cycle metabolic pathway, as determined for mesocarp tissue of fruits of high-yielding palms plants versus mesocarp tissue of fruits of low-yielding palm plants.
  • FIGS. 6A-B and 7 provide heat map data and graphs of relative concentration versus time post-pollination for various metabolites of amino acid metabolic pathways
  • FIGS. 8 and 9 provide heat map data and graphs of relative concentration versus time post-pollination for various metabolites of purine and pyrimidine metabolic pathways
  • FIGS. 10 and 11 provide heat map data and graphs of relative concentration versus time post-pollination for various metabolites of oxidative phosphorylation metabolic pathways, i.e. electron transport chain and ATP synthesis
  • FIGS. 12 and 13 provide heat map data and graphs of relative concentration versus time post-pollination for various metabolites of glycolytic pathways, all as determined for mesocarp tissue of fruits of high-yielding palms plants versus mesocarp tissue of fruits of low-yielding palm plants.
  • increasing darkness of shading indicates increasingly higher levels of a given metabolite.
  • time is indicated on the x- axis as weeks post-pollination, data for high-yielding palms are indicated as solid lines, and data for low-yielding palms are indicated as dashed lines.
  • TABLE 2 lists thirty-six metabolites useful for identifying high-yielding palms and predicting palm oil yields of test palms based on the metabolites being present at higher levels in mesocarp tissue of fruits of high-yielding oil palm plants in comparison to low-yielding oil palm plants.
  • the results indicate that levels of 2-aminobutyric acid, 2- hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, arginine, asparagine, cadaverine, cyclohexylamine, cytidine, fructose- 1 ,6-diphosphate, gamma-aminobutyric acid, glucose- 1 -phosphate, glycerol-3 -phosphate, glyoxylic acid, guanosine, homo-L-arginine, glutamine, indole-3-acetaldehyde, lysine, malic acid, myo- inositol-phosphate (i.e.
  • TABLE 3 lists thirty-five metabolites useful for identifying high- yielding palms and predicting palm oil yields of test palms based on the metabolites being present at lower levels in mesocarp tissue of high-yielding oil palm plants versus low-yielding oil palm plants.
  • the results indicate that levels of 1- methyladenosine, 2-oxoglutaric acid, 3-phosphoglyceric acid, 4-aminophenylsulfone, 4- guanidinobutyric acid, 6,8-thioctic acid, ascorbic acid, adenosine triphosphate, chelidonic acid, citramalic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, isocitric acid, methyl sulfate,
  • metronidazole metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N-acetylhistidine, N- methylproline, pantothenic acid, phosphorylcholine, quinic acid, ribulose-5-phosphate, symmetric dimethyl arginine, stachydrine, tartaric acid, tetrahydrouridine, thiamine, tryptophan, and uridine triphosphate were at least 5% lower in mesocarp tissue of fruits of high- yielding palms compared to mesocarp tissue of fruits of low-yielding palms, also for one or more weeks from 12 to 22 weeks post-pollination.
  • a high- yielding oil palm plant can be obtained from a dura x pisifera cross, e.g. a dura x pisifera cross that is likely to segregate in terms of palm-oil yield performance or for commercial production of palm-oil producing plants, as described in the following prophetic example.
  • a dura x pisifera cross is carried out by standard methods, and progeny are obtained.
  • the progeny are field planted and grown, until fruiting, for at least 18 months.
  • Fruit bunch specimens are taken from each progeny plant for testing at one or more time points from 1 1 to 23 weeks post-pollination. Testing at multiple time point is preferable to testing at a single time point, as the former provides a higher number of comparative points.
  • Fruitlets are separated immediately from the bunch and ten fruits samples are randomly selected therefrom.
  • Mesocarp tissue is obtained from the ten fruit samples and combined into one mesocarp tissue sample per progeny plant. The resulting mesocarp tissue samples are frozen under liquid nitrogen until further processing.
  • the mesocarp tissue samples are subjected to extraction as described in Example 1 above.
  • other mild polar extraction conditions known to those skilled in the art can be used, e.g. methanol or ethanol extraction, with or without a proportion of water.
  • Non-polar components are removed by subsequent solvent partitioning with a non-polar solvent, e.g. chloroform,
  • non-polar components can be removed by solid phase extraction using a non-polar stationary phase, e.g. CI 8. Also alternatively, non-polar components can be left in the mixture. The samples are then evaporated to dryness.
  • Samples are then prepared for analysis by any of various standard approaches that are well known in the art by reconstituting the samples to the same concentration in a solvent system appropriate for the approach.
  • HPLC and LC-MS can be used for analysis, in which case samples can be reconstituted in methanol-water mixtures or dimethyl sulfoxide, with or without pH modifier added.
  • GC-MS derivatization can be used, in which case samples can be reconstituted as described in Example 1 or by various approaches known in the art.
  • NMR can be used, in which case samples can be reconstituted in a deuterated solvent such as methanol-iit or dimethyl sulfoxide-c3 ⁇ 4-
  • the levels of one or more metabolites in the samples are then determined.
  • the metabolites are selected from the group consisting of 2-aminobutyric acid, 2- hydroxyvaleric acid, adenine, asymmetric dimethyl arginine, alanine, alanyl-alanine, argi ine, asparagine, cadaverine, cyclohexylamine, cytidtne, gamma-aminobutyric acid, glyoxylic acid, guanosine, homo-L-arginine, glutamine, indole-3-acetaldehyde, lysine, myo-inositol-phosphate, myo-inositol-2-phosphate, nicotinamide, ⁇ - ⁇ -methylarginine, octylamine, ophthalmic acid, serine, spermidine, triethanolamine, uracil, uridine, and beta-alanine.
  • the metabolites are selected from the group consisting of 1- methyladenosine, 4-aminophenylsulfone, 4-guanidinobutyric acid, 6,8-thioctic acid, ascorbic acid, chelidonic acid, cytosine, etidronic acid, glucaric acid, glutathione disulfide, imidazole-4-acetic acid, imidazolelactic acid, methyl sulfate, metronidazole, mucic acid, N6-acetyllysine, N6-methyladenine, N-acetylhistidine, N-methylproline, pantothenic acid, phosphorylcholine, quinic acid, symmetric dimethyl arginine, stachydrine, tartaric acid, tetrahydrouridine, thiamine, and tryptophan.
  • the metabolites are selected from the group consisting of adenosine triphosphate and uridine triphosphate.
  • the metabolites are selected from the
  • the difference can be that the level of the metabolite in the sample is at least 15% higher than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the difference can be that the level of the metabolite in the sample is at least 5% lower than the level of the metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the difference can be that the level of adenine is at least 15% higher in the sample in comparison to the level of adenine in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the progeny plants are classified in terms of palm oil yields, resulting in a distribution of progeny spanning a spectrum of high-yielding palm plants to low-yielding palm plants.
  • the set of high-yielding progeny are then selected for further breeding stock development.
  • progeny for which the level of adenine is at least 15% higher in the sample in comparison to the level of adenine in the mesocarp tissue of the fruit of the reference oil palm plant can be classified as having a high oil palm yield.
  • progeny can then be selected for further breeding stock development.
  • Parental oil palm plants that are suitable for producing high- yielding progeny can be identified by use of metabolites as markers for high oil-palm yields as described in the following prophetic example.
  • Two dura x pisifera crosses are carried out by standard methods, and progeny are obtained.
  • the two crosses can include one parent in common, e.g. a particular individual dura plant is used for the two crosses, or alternatively a particular individual pisifera plant is used for the two crosses. Alternatively, the two crosses can include no parent in common.
  • the progeny are field planted and grown, fruit bunch specimens are taken, and mesocarp tissue samples are obtained and tested, as described in Example 3, above.
  • the concentrations of one or more metabolites are then determined in the mesocarp tissue samples obtained from the progeny of both crosses, and compared with respect to the metabolites in the mesocarp tissue of the fruit of the reference oil palm plant, also as described in Example 3, above.
  • the parental plants that yield the cross that exhibits relative metabolite concentrations consistent with higher yields of palm oil, e.g. higher levels of adenine, can then selected for further crosses, e.g. with other parental plants that also yield crosses that exhibit relative metabolite concentrations consistent with higher yields of palm oil, in order to obtain high-yielding oil palm plants.
  • the metabolites and methods described in the Examples above can also be used for breeding stock improvement (e.g. comparisons or selections), estate management practices, selection of high yielding wild material with other potentially useful traits for inclusion into breeding programmes, and selection of high yielding ortets for tissue culture production markers, as described in the following prophetic example.
  • Fruit mesocarp samples are collected from groups of palm-oil producing palm plants of similar age and are analyzed as described in Example 3.
  • the groups of palm samples can be selected to compare, for example, two different commercial palm-oil producing populations, a commercial palm-oil producing population and wild material, or palms grown from tissue culture processes.
  • Metabolite markers are analyzed and used to compare the potential palm-oil yields of materials, as described in Example 3. Based on the results, individual palm plants exhibiting high yields are selected and used for breeding stock improvement, for estate management practices, inclusion in breeding programmes, and/or selection of high yielding ortets for tissue culture production.
  • a high-yielding oil palm plant can be obtained by use of the methods described in the Examples above, wherein one or more metabolites are detected by, for example, CE- MS, LC-MS, GC-MS, simple HPLC analysis, NMR analysis, an enzyme-based biosensor, or a bioluminescence kit, with the choice of detection depending on the metabolite, as described in the following prophetic example.
  • Oil palm plants are obtained for analysis as described in the Examples above.
  • oil palm plants can be classified as high-yielding in comparison to reference plants, with the oil palm plants and the reference plants each being considered either individually or in groups.
  • individual comparisons i.e. comparisons involving single plants
  • multiple fruit bunch and fruit samples should be collected and analyzed, e.g. preferably greater than four samples per individual, and more preferably greater than seven samples per individual.
  • group samples i.e. comparisons involving multiple plants, multiple plant specimens should also be collected within each group.
  • Extraction can be conducted using any of the various extraction methods described in the Examples above. Alternatively, additional methods known to those skilled in the art can be used, e.g. super critical fluid extraction.
  • One or more of the metabolites noted in Example 3 can be used, alone or in combination, to identify parental oil palm plants suitable for producing high-yielding progeny or to classify high-yielding oil palm plants, both relative to reference oil palm plants. Multiple metabolites can be used in particular to reduce the potential effect of experimental error.
  • the levels of the one or more metabolites in the mesocarp tissue of the fruit of a parental or test oil palm plant can be determined by, for example, CE-MS, LC-MS, GC-MS, simple HPLC analysis, NMR analysis, an enzyme-based biosensor, or a bioluminescence kit. Brief descriptions of quantitation methods useful for various specific metabolites follow.
  • parental oil palm plants suitable for producing high-yielding progeny can be identified, and test oil palm plants that are high-yielding can be classified, both relative to reference oil palm plants, based on the level of adenine in the mesocarp tissue of the fruit of the parental or test oil palm plant being at least 15% higher, e.g. between 110% and 240% higher, than the level of adenine in the mesocarp tissue of the fruit of the reference oil palm plant, 1 1 to 23 weeks, e.g. 1 1 to 21 weeks, after pollination thereof.
  • Adenine can be detected and quantified efficiently using either CE-MS or LC-MS methods as described above.
  • Adenine can also be detected and quantified by simple HPLC analysis and quantitation using reversed phase methods, e.g. using a CI 8 column and a solvent gradient from water (0.1% formic acid or 0.05% trifluoroacetic acid) to acetonitrile (0.1% formic acid or 0.05% trifluoroacetic acid) and UV detection at 254 nm, as is known in the art.
  • a calibration curve can be prepared to accurately determine sample concentrations using an authentic standard of adenine and determining peak area of at least 5 different concentrations between 0.001 mg/ml to 0.1 mg/ml.
  • Extract concentrations should be prepared to 10 mg/ml to allow sufficient peak size for accurate concentration determination.
  • NMR analysis can also be used to efficiently compare adenine in biological samples, e.g. based on identification and quantitative analysis of plant metabolites in tissue extracts, as taught for example by Krishnan et al., Metabolite Fingerprinting and Profiling in Plants Using NMR, 56 Journal of Experimental Botany 255-265 (2005).
  • parental oil palm plants suitable for producing high-yielding progeny can be identified, and test oil palm plants that are high-yielding can be classified, both relative to reference oil palm plants, based on the level of ascorbic acid in the mesocarp tissue of the fruit of the parental or test oil palm plant being at least 5% lower, e.g. between 22% to 57% lower, than the level of ascorbic acid in the mesocarp tissue of the fruit of the reference oil palm plant, 11 to 23 weeks, e.g. 13 to 21 weeks, after pollination thereof.
  • Ascorbic acid can be detected and quantified efficiently using CE-MS, as described above. Ascorbic acid also can be detected and quantified by use of an enzyme- based biosensor, e.g. horseradish peroxidase immobilized on the surface of an ion sensitive field effect transistor, as taught for example by Volotovsky et al., Ascorbic Acid Determination with an Ion-Sensitive Field Effect Transistor-Based Peroxidase Biosensor, 359 Analytica Chimica Acta 143-148 (1998). Ascorbic acid also can be detected and quantified by enzymatic oxidation followed by spectrophotometric monitoring, e.g.
  • parental oil palm plants suitable for producing high-yielding progeny can be identified, and test oil palm plants that are high-yielding can be classified, both relative to reference oil palm plants, based on the level of adenosine triphosphate in the mesocarp tissue of the fruit of the parental or test oil palm plant being at least 5% lower, e.g. between 33% to 55% lower, than the level of ascorbic acid in the mesocarp tissue of the fruit of the reference oil palm plant, 1 1 to 23 weeks, e.g. 1 1 to 21 weeks, after pollination thereof.
  • Adenosine triphosphate can be detected and quantified efficiently using either CE-MS or LC-MS methods as described above. Adenosine triphosphate also can be detected and quantified by HPLC analyses that are known in the art. Adenosine triphosphate also can be detected and quantified by NMR methods, e.g. based on the characteristic NMR signal patterns observed for adenosine triphosphate in vitro, as taught for example by Bligny et al., NMR and Plant Metabolism, 4 Current Opinion in Plant Biology 191 (2001). Adenosine triphosphate can also be detected and quantified by use of a bioluminescence kit, e.g. based on use of an ATP Determination Kit (Molecular Probes, p/n: A22066) for quantitative determination of ATP with recombinant firefly luciferase and its substrate D-luciferin.
  • a bioluminescence kit e.g. based on use of an ATP Determination Kit (
  • the methods and kits disclosed herein are useful for obtaining high-yielding oil palms and for predicting oil yields of test oil palm plants, and thus for improving commercial production of palm oil.
  • TABLE 1 Subset of metabolites of interest identified based on being present at relatively high levels in mesocarp tissue of fruits of high-yielding oil palm plants ("high in high-yielding plants"), or alternatively low-yielding oil palm plants ("high in low- yielding plants”), at 16 weeks post-pollination.

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CN105974038A (zh) * 2016-06-24 2016-09-28 广西灵峰药业有限公司 一种无蔗糖益母草颗粒的生产质量控制方法
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Publication number Priority date Publication date Assignee Title
WO2015034345A1 (en) * 2013-09-04 2015-03-12 Sime Darby Malaysia Berhad Methods for predicting oil yield of a test oil palm plant
WO2015034344A1 (en) * 2013-09-04 2015-03-12 Sime Darby Malaysia Berhad Methods for predicting oil yield of a test oil palm plant
CN105911187A (zh) * 2016-04-25 2016-08-31 广西壮族自治区梧州食品药品检验所 液质串联检测妇康宁片中盐酸水苏碱含量的方法
CN106008432A (zh) * 2016-06-12 2016-10-12 王伟明 从桄榔果实中制备白屈菜酸盐的方法
CN105974038A (zh) * 2016-06-24 2016-09-28 广西灵峰药业有限公司 一种无蔗糖益母草颗粒的生产质量控制方法
WO2022235147A1 (en) * 2021-05-06 2022-11-10 Universiti Kebangsaan Malaysia A method for classifying oil palm fruit ripeness

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