WO2013089558A1 - Procédés d'obtention de palmiers à huile avec un rendement élevé - Google Patents

Procédés d'obtention de palmiers à huile avec un rendement élevé Download PDF

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Publication number
WO2013089558A1
WO2013089558A1 PCT/MY2012/000242 MY2012000242W WO2013089558A1 WO 2013089558 A1 WO2013089558 A1 WO 2013089558A1 MY 2012000242 W MY2012000242 W MY 2012000242W WO 2013089558 A1 WO2013089558 A1 WO 2013089558A1
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WIPO (PCT)
Prior art keywords
oil palm
level
fruit
metabolite
palm plant
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PCT/MY2012/000242
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English (en)
Inventor
Teh Huey FANG
May Hong Ping LI
Neoh Bee KEAT
Jaime Low Yoke SUM
Nalisha Binti ITHNIN
Theresa NG Lee MEI
Thang Yin MEE
David Ross APPLETON
Yusof@Hassan HIRZUN BIN MOHD
Kulaveerasingam HARIKRISHNA A/L
Mohamed MOHAIMI
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Sime Darby Malaysia Berhad
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Priority to BR112014014737A priority Critical patent/BR112014014737A2/pt
Priority to SG11201403226SA priority patent/SG11201403226SA/en
Publication of WO2013089558A1 publication Critical patent/WO2013089558A1/fr
Priority to CR20140281A priority patent/CR20140281A/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
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
    • 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
    • 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
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy

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 first metabolite and the level of a second metabolite in mesocarp tissue of a fruit of a parental oil palm plant 1 1 to 21 weeks after pollination thereof.
  • the first metabolite is selected from the group consisting of malic acid and pyruvic acid.
  • the second metabolite is selected from the group consisting of 2- oxoglutaric acid, isocitric acid, and citramalic acid.
  • the method also comprises determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 1 1 to 21 weeks after pollination thereof.
  • the difference is that the level of the first metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 5% lower than the level of the second 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 first metabolite and the level of a second metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 21 weeks after pollination thereof.
  • the first metabolite is selected from the group consisting of malic acid and pyruvic acid.
  • the second metabolite is selected from the group consisting of 2- oxoglutaric acid, isocitric acid, and citramalic acid.
  • the method also comprises determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 21 weeks after pollination thereof.
  • the difference is that the level of the first metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 15% higher than the level of the first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and the level of the second metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 5% lower than the level of the second 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 first reagent for quantitative detection of a first metabolite.
  • the first metabolite is selected from the group consisting of malic acid and pyruvic acid.
  • the kit also comprises a second reagent for quantitative detection of a second metabolite.
  • the second metabolite is selected from the group consisting of 2- oxoglutaric acid, isocitric acid, and citramalic acid.
  • the kit also comprises an extract of a mesocarp tissue of a fruit of a reference oil palm plant 11 to 21 weeks after pollination thereof.
  • the kit also comprises instructions indicating use of the first reagent and the second reagent for determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 11 to 21 weeks after pollination thereof, and the level of the first metabolite and the level of the second metabolite in the extract of the mesocarp tissue of the fruit of the reference oil palm plant.
  • the level of the first 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 first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant
  • the level of the second 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 second metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the method comprises determining the level of a first metabolite and the level of a second metabolite in mesocarp tissue of a fruit of a parental oil palm plant 11 to 21 weeks after pollination thereof.
  • the first metabolite is selected from the group consisting of fructose- 1 ,6-diphosphate, glucose- 1 -phosphate, glycerol-3-phosphate, pyruvic acid, and ribose-5-phosphate.
  • the second metabolite is selected from the group consisting of 3- phosphoglyceric acid and ribulose-5-phosphate.
  • the method also comprises determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 1 1 to 21 weeks after pollination thereof.
  • the difference is that the level of the first metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 5% lower than the level of the second 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 first metabolite and the level of a second metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 21 weeks after pollination thereof.
  • the first metabolite is selected from the group consisting of fructose- 1 ,6-diphosphate, glucose- 1 -phosphate, glycerol-3-phosphate, pyruvic acid, and ribose-5-phosphate.
  • the second metabolite is selected from the group consisting of 3- phosphoglyceric acid and ribulose-5-phosphate.
  • the method also comprises determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also I I to 21 weeks after pollination thereof.
  • the difference is that the level of the first metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 15% higher than the level of the first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and the level of the second metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 5% lower than the level of the second 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 first reagent for quantitative detection of a first metabolite.
  • the first metabolite is selected from the group consisting of fructose- 1 ,6-diphosphate, glucose- 1 -phosphate, glycerol-3 -phosphate, pyruvic acid, and ribose-5-phosphate.
  • the kit also comprises a second reagent for quantitative detection of a second metabolite.
  • the second metabolite is selected from the group consisting of 3 -phosphoglyceric acid and ribulose-5-phosphate.
  • the kit also comprises an extract of a mesocarp tissue of a fruit of a reference oil palm plant 1 1 to 21 weeks after pollination thereof.
  • the kit also comprises instructions indicating use of the first reagent and the second reagent for determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 1 1 to 21 weeks after pollination thereof, and the level of the first metabolite and the level of the second metabolite in the extract of the mesocarp tissue of the fruit of the reference oil palm plant.
  • the level of the first 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 first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant
  • the level of the second 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 second 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
  • 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. 6 A 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 glycolytic 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 levels of a first metabolite and a second 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.
  • First and second metabolites useful in this regard include (i) malic acid and pyruvic acid, and (ii) 2-oxoglutaric acid, isocitric acid, and citramalic acid, respectively.
  • Additional first and second metabolites useful in this regard include (i) fructose- 1 ,6-diphosphate, glucose- 1- phosphate, glycerol-3-phosphate, pyruvic acid, and ribose-5-phosphatei and (ii) 3- phosphoglyceric acid and ribuIose-5 -phosphate, respectively.
  • the application provides methods for obtaining high-yielding oil palm plants comprising determining the levels of one of each of the above-noted first and second metabolites in mesocarp tissue of a fruit of a parental oil palm plant 1 1 to 21 weeks after pollination thereof, determining whether there " is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 21 weeks after pollination thereof, wherein the difference is that (a) the level of the first metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and (b) the level of the second metabolite in the mesocarp tissue
  • the application provides methods for predicting oil yield of test oil palm plants comprising determining the levels of one of each of the above-noted first and second metabolites in mesocarp tissue of a fruit of a test oil palm plant 11 to 21 weeks after pollination thereof, determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 21 weeks after pollination thereof, wherein the difference is that (a) the level of the first metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 15% higher than the level of the first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and (b) the level of the second metabolite in the mesocarp tissue of the fruit of
  • kits for obtaining a high-yielding oil palm plant or predicting oil yield of a test oil palm plant comprising a first reagent for quantitative detection of one of the above-noted first metabolites, a second reagent for quantitative detection of one of the above-noted second metabolites, an extract of a mesocarp tissue of a fruit of a reference oil palm plant 11 to 21 weeks after pollination thereof; and instructions indicating use of the first and second reagents for determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 11 to 21 weeks after pollination thereof, and the level of the first metabolite and the level of the second metabolite in the extract of the mesocarp tissue of the fruit of the reference oil palm plant, wherein the difference is that (a) the level of the first metabolite in the mesocarp tissue of the fruit
  • 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 refers 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 first metabolite and the level of a second metabolite in mesocarp tissue of a fruit of a parental oil palm plant 1 1 to 21 weeks after pollination thereof, (ii) determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 1 1 to 21 weeks after pollination thereof, wherein the difference is that (a) the level of the first metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and (b)
  • the first metabolite is selected from the group consisting of malic acid and pyruvic acid
  • the second metabolite is selected from the group consisting of 2-oxoglutaric acid, isocitric acid, and citramalic acid.
  • These metabolites are associated with the citric acid cycle, which plays a central role in metabolism by providing biosynthetic precursors and sources of energy for many metabolic pathways. Strikingly, although fatty acid biosynthesis and triacylglycerol biosynthesis require biosynthetic precursors and sources of energy, it would not have been apparent whether levels of each of the first metabolites, i.e. malic acid and pyruvic acid, and of each of the second metabolites, i.e.
  • 2-oxoglutaric acid, isocitric acid, and citramalic acid, 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. whether higher levels of the first and second metabolites might drive production of fatty acids and triacylglycerols, or whether the levels may have no effect on, and be unaffected by, production of fatty acids and triacylglycerol, or whether the levels may be reduced because of increased production of fatty acids and triacylglycerols, or some combination thereof.
  • the level of the above-noted first metabolite and the level of the above-noted second metabolite 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 levels of the first and second metabolites 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. 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.
  • 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 21 weeks after pollination thereof, i.e. after the pollination event from which the fruit originated.
  • the mesocarp tissue may be obtained and tested 1 1-19 weeks, 1 1-17 weeks, 11-15 weeks, 11-13 weeks, 13-21 weeks, 13-19 weeks, 13-17 weeks, 13-15 weeks, 15-21 weeks, 15-19 weeks, 15-17 weeks, 17-21 weeks, 17-19 weeks, 19-21 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, or 21 weeks after pollination thereof.
  • the step of determining the levels of the first and second metabolites 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 levels of the first and second metabolite in mesocarp tissue of a fruit of a parental oil palm plant may be carried out by determining (i) the mean level of the first metabolite in mesocarp tissue of at least one fruit of at least one parental oil palm plant, and (ii) the mean level of the second metabolite in mesocarp tissue of the at least one fruit of the at least one parental oil palm plant.
  • the mean levels of the first and second metabolites may be determined, for example, by determining the levels of the first 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 first metabolite, and applying the same approach with respect to the second metabolite to obtain the mean level of the second metabolite.
  • the mean level of the first metabolite may be determined by determining the levels of the first 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 first metabolite, and applying the same approach with respect to the second metabolite to obtain the mean level of the second metabolite.
  • the mean levels of the first and second metabolites may be determined by determining the levels of the first 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 first metabolite, and applying the same approach with respect to the second metabolite to obtain the mean level of the second metabolite.
  • the mean levels of the first and second metabolites may be determined by combining mesocarp tissue from at least one fruit of at least one parental oil palm plant and then determining the levels of the first and second metabolites in the combined mesocarp tissue to obtain the mean levels of the first and second metabolites.
  • the mean levels of the first and second metabolites may be determined by various combinations of the above-noted approaches.
  • levels are determined for more than one of each of the first and second metabolites.
  • levels are determined for more than one of the first metabolites, i.e. for both malic acid and pyruvic acid, as well as for at least one of the second metabolites, i.e. 2-oxoglutaric acid, isocitric acid, and/or citramalic acid.
  • levels are determined for at least one of the first metabolites, i.e. for at least one of malic acid and pyruvic acid, as well as for more than one of the second metabolites, i.e. for two or more of 2-oxoglutaric acid, isocitric acid, and/or citramalic acid.
  • determining the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is necessarily transformative of the mesocarp tissue.
  • the levels cannot be determined, for example, merely based on appearance of the mesocarp tissue. Rather, determination of the levels of the first and second metabolites in the mesocarp tissue requires physical disruption and/or chemical treatment of the mesocarp tissue.
  • the levels of the first and second metabolites 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 first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a reference oil palm plant may be carried out by determining the levels of the first and second metabolites 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 between those levels and the levels of the first and second metabolites in the mesocarp tissue of the fruit of the parental oil palm plant.
  • 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. 1 1 to 21 weeks after pollination thereof, or 1 1-19 weeks, 11- 17 weeks, 1 1-15 weeks, 1 1-13 weeks, 13-21 weeks, 13-19 weeks, 13-17 weeks, 13-15 weeks, 1 -21 weeks, 15-19 weeks, 15-17 weeks, 17-21 weeks, 17-19 weeks, 19-21 weeks, 1 1 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, or 21 weeks after pollination thereof.
  • the levels of the first and second metabolites in the mesocarp tissue of the fruit of the parental oil palm plant may be determined as mean levels of the first and second metabolites, respectively.
  • the levels of the first and second metabolites in mesocarp tissue of a fruit of a reference oil palm plant may be determined before, during, or after determination of the levels of the first and second metabolites 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 (a) the level of the first metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the first 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, and (b) the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 5% lower than the level of the second 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 0.95: 1 or lower, and wherein the p value for the
  • such a comparison is considered to reveal a biologically and/or statistically significant difference based on the difference being that the level of the first 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, and wherein the level of the second 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
  • the difference relates to a mean level of the first metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and/or the reference oil palm plant, and/or a mean level of the second 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 levels of the first and second metabolites in the mesocarp tissue of a fruit of the parental oil palm plants are higher than the mean levels of the first and second metabolites, respectively, in the mesocarp tissue of at least one fruit of at least one reference oil palm plant.
  • the difference is that the mean levels of the first and second metabolites in the mesocarp tissue of at least one fruit of at least one parental oil palm plant is higher than the levels of the first and second metabolites, respectively, in the mesocarp tissue of a fruit of the reference oil palm plant.
  • the difference is that the mean levels of the first and second metabolites in the mesocarp tissue of at least one fruit of at least one parental oil palm plant is higher than the mean levels of the first and second metabolites in the mesocarp tissue of at least one fruit of at least one reference oil palm plant.
  • the difference is that the mean levels of the first and second metabolites in the mesocarp tissue of a plurality of the fruits of a plurality of the parental oil palm plants is higher than the mean levels of the first and second metabolites, respectively, 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 first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the malic acid in the mesocarp tissue of the fruit of the parental oil palm plant 15 to 17 weeks after pollination thereof is 43% to 100% higher than the level of the malic acid in the mesocarp tissue of the fruit of the reference oil palm plant 15 to 17 weeks after pollination thereof, and the level of the 2-oxoglutaric acid in the mesocarp tissue of the fruit of the parental oil palm plant 1 1 to 15 weeks after pollination thereof is 50% to 69% lower than the level of the 2-oxoglutaric 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 first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the malic acid in the mesocarp tissue of the fruit of the parental oil palm plant 15 to 17 weeks after pollination thereof is 43% to 100% higher than the level of the malic acid in the mesocarp tissue of the fruit of the reference oil palm plant 15 to 17 weeks after pollination thereof, and the level of the isocitric acid in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 19 weeks after pollination thereof is 13% to 35% lower than the level of the isocitric acid in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 19 weeks after pollination thereof.
  • the difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the malic acid in the mesocarp tissue of the fruit of the parental oil palm plant 5 to 17 weeks after pollination thereof is 43% to 100% higher than the level of the malic acid in the mesocarp tissue of the fruit of the reference oil palm plant 15 to 17 weeks after pollination thereof, and the level of the citramalic acid in the mesocarp tissue of the fruit of the parental oil palm plant 17 to 19 weeks after pollination thereof is 30% to 45% lower than the level of the citramalic acid 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 first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the pyruvic acid in the mesocarp tissue of the fruit of the parental oil palm plant 1 1 to 13 weeks after pollination thereof is 33% to 82% higher than the level of the pyruvic acid in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 13 weeks after pollination thereof, and the level of the 2-oxoglutaric acid in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 15 weeks after pollination thereof is 50% to 69% lower than the level of the 2-oxoglutaric 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 first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the pyruvic acid in the mesocarp tissue of the fruit of the parental oil palm plant 1 1 to 13 weeks after pollination thereof is 33% to 82% higher than the level of the pyruvic acid in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 13 weeks after pollination thereof, and the level of the isocitric acid in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 19 weeks after pollination thereof is 13% to 35% lower than the level of the isocitric acid in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 19 weeks after pollination thereof.
  • the difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the pyruvic acid in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 13 weeks after pollination thereof is 33% to 82% higher than the level of the pyruvic acid in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 13 weeks after pollination thereof, and the level of the citramalic acid in the mesocarp tissue of the fruit of the parental oil palm plant 17 to 19 weeks after pollination thereof is 30% to 45% lower than the level of the citramalic acid 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 first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a 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 first metabolites, i.e.
  • 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 first metabolites, i.e. malic acid and pyruvic acid, and/or more than one of the second metabolites, i.e. 2-oxoglutaric acid, isocitric acid, and citramalic acid, 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 first metabolites i.e. malic acid and pyruvic acid
  • the second metabolites i.e. 2-oxoglutaric acid, isocitric acid, and citramalic acid
  • the step of selecting is based on differences with respect to two of the first metabolites and one of the second metabolites, or one of the first metabolites and two of the second metabolites. Also for example, in some embodiments the step of selecting is based on differences with respect to combination of one or two of the first metabolites, and one, two, or three of the second 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.
  • the method comprises (i) determining the level of a first metabolite and the level of a second metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 21 weeks after pollination thereof, (ii) determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 1 1 to 21 weeks after pollination thereof, wherein the difference is that the level of the first metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 15% higher than the level of the first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and the level of the second metabolite
  • the metabolites noted above as being useful in the method for obtaining a high- yielding oil palm plant i.e. a first metabolite selected from the group consisting of malic acid and pyruvic acid, and a second metabolite selected from the group consisting of 2- oxoglutaric acid, isocitric acid, and citramalic acid, are also useful in the method for predicting oil yield of a test oil palm plant.
  • the step of determining the level of the above-noted first metabolite and the level of the above-noted second metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 21 weeks after pollination thereof may also be carried out similarly as described above, e.g.
  • 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 first and/or second metabolites, except that the levels of the first and second metabolites 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.
  • the step of determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the first metabolite and the level of the second 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 (a) the level of the first metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the first 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 less than 0.1, and (b) the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 5% lower than the level of the second metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, where
  • the difference may be based on any of the specific differences noted above with respect to each of the first and second metabolites, e.g. in some embodiments the difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the malic acid in the mesocarp tissue of the fruit of the parental oil palm plant 15 to 17 weeks after pollination thereof is 43% to 100% higher than the level of the malic acid in the mesocarp tissue of the fruit of the reference oil palm plant 15 to 17 weeks after pollination thereof, and the level of the 2-oxoglutaric acid in the mesocarp tissue of the fruit of the parental oil palm plant 1 1 to 15 weeks after pollination thereof is 50% to 69% lower than the level of the 2-oxoglutaric acid in the mesocar
  • the predicting step may be carried out, for example, based on the amount of the difference in the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant, and/or based on correlations between levels of the metabolites 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 each of the first and second metabolites. Kit for Obtaining a High- Yielding Oil Palm Plant
  • kits for obtaining a high-yielding oil palm plant or predicting oil yield of a test oil palm plant comprises: (i) a first reagent for quantitative detection of a first metabolite; (ii) a second reagent for quantitative detection of a second metabolite; (iii) an extract of a mesocarp tissue of a fruit of a reference oil palm plant 11 to 21 weeks after pollination thereof; and (iv) instructions indicating use of the first reagent and the second reagent for determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 11 to 21 weeks after pollination thereof, and the level of the first metabolite and the level of the second 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 first reagent for quantitative detection of a first metabol
  • the metabolites described above as being useful in the method for obtaining a high-yielding oil palm plant i.e. a first metabolite selected from the group consisting of malic acid and pyruvic acid, and a second metabolite selected from the group consisting of 2-oxoglutaric acid, isocitric acid, and citramalic acid, are also useful in the kit for obtaining a high-yielding oil palm plant.
  • the first reagent for quantitative detection of the first metabolite and the second reagent for quantitative detection of the second 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 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.
  • the method comprises: (i) determining the level of a first metabolite and the level of a second metabolite in mesocarp tissue of a fruit of a parental oil palm plant 1 1 to 21 weeks after pollination thereof; (ii) determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 21 weeks after pollination thereof, wherein the difference is that the level of the first metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and the level of the second metabol
  • the first metabolite is selected from the group consisting of fructose- 1,6-diphosphate, glucose- 1 -phosphate, glycerol-3-phosphate, pyruvic acid, and ribose-5-phosphate
  • the second metabolite is selected from the group consisting of 3-phosphoglyceric acid and ribulose-5-phosphate.
  • 3-phosphoglyceric acid and ribulose-5-phosphate, 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. whether higher levels of the first and second metabolites might drive production of fatty acids and tnacylglycerols, or whether the levels may have no effect on, and be unaffected by, production of fatty acids and triacylglycerol, or whether the levels may be reduced because of increased production of fatty acids and tnacylglycerols, or some combination thereof.
  • the level of the first metabolite i.e.
  • the level of the second metabolite, i.e. 3-phosphoglyceric acid and ribulose-5-phosphate, in mesocarp tissue of a fruit of a parental oil palm plant may also be determined as described above.
  • the levels 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. 11 to 21 weeks after pollination thereof, for example, 11-19 weeks, 11-17 weeks, 1 1-15 weeks, 11-13 weeks, 13-21 weeks, 13-19 weeks, 13-17 weeks, 13-15 weeks, 15-21 weeks, 15-19 weeks, 15-17 weeks, 17-21 weeks, 17-19 weeks, 19-21 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, or 21 weeks after pollination thereof.
  • the step of determining the level of the first metabolite and the level of the second 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 levels of the first and second metabolite in mesocarp tissue of a fruit of a parental oil palm plant may be carried out by determining (i) the mean level of the first metabolite in mesocarp tissue of at least one fruit of at least one parental oil palm plant, and (ii) the mean level of the second metabolite in mesocarp tissue of the at least one fruit of the at least one parental oil palm plant.
  • the mean levels of the first and second metabolites may be determined, for example, by determining the levels of the first 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 first metabolite, and applying the same approach with respect to the second metabolite to obtain the mean level of the second metabolite.
  • the mean level of the first metabolite may be determined by determining the levels of the first 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 first metabolite, and applying the same approach with respect to the second metabolite to obtain the mean level of the second metabolite.
  • the mean levels of the first and second metabolites may be determined by determining the levels of the first 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 first metabolite, and applying the same approach with respect to the second metabolite to obtain the mean level of the second metabolite.
  • the mean levels of the first and second metabolites may be determined by combining mesocarp tissue from at least one fruit of at least one parental oil palm plant and then determining the levels of the first and second metabolites in the combined mesocarp tissue to obtain the mean levels of the first and second metabolites.
  • the mean levels of the first and second metabolites may be determined by various combinations of the above-noted approaches.
  • levels are determined for more than one of each of the first and second metabolites.
  • levels are determined for more than one of the first metabolites, i.e. fructose- 1,6-diphosphate, glucose- 1 -phosphate, glycerol-3 -phosphate, pyruvic acid, and ribose-5-phosphate, as well as for at least one of the second metabolites, i.e. 3-phosphoglyceric acid and ribulose-5- phosphate.
  • levels are determined for at least one of the first metabolites, i.e.
  • fructose- 1 ,6-diphosphate for at least one of fructose- 1 ,6-diphosphate, glucose- 1 - phosphate, glycerol-3-phosphate, pyruvic acid, and ribose-5 -phosphate, as well as for more than one of the second metabolites, i.e. for both 3-phosphoglyceric acid and ribulose-5 -phosphate .
  • determining the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is necessarily transformative of the mesocarp tissue, and the levels of the metabolites 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 first metabolite, i.e. fructose- 1 ,6-diphosphate, glucose- 1 -phosphate, glycerol-3-phosphate, pyruvic acid, and ribose-5-phosphate, and the level of the second metabolite, i.e. 3-phosphoglyceric acid and ribulose-5-phosphate, in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second 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 metabolites, 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 checking for a difference therebetween.
  • 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 checking for a difference therebetween.
  • the mesocarp tissue of the fruit of the parental oil palm plant the mesocarp tissue of the fruit of the reference oil palm plant may be obtained and
  • 1 1 to 21 weeks after pollination thereof for example 1 1-19 weeks, 1 1-17 weeks, 1 1-15 weeks, 11-13 weeks, 13-21 weeks, 13-19 weeks, 13-17 weeks, 13-15 weeks, 15-21 weeks, 15-19 weeks, 15-17 weeks, 17-21 weeks, 17-19 weeks, 19-21 weeks, 1 1 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, or 21 weeks after pollination thereof.
  • the levels of the first and second metabolites in the mesocarp tissue of the fruit of the parental oil palm plant may be determined as mean levels.
  • the levels of the first and second metabolites in mesocarp tissue of a fruit of a reference oil palm plant may be determined before, during, or after determination of the levels of the first and second metabolites 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 (a) the level of the first metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the first 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, and (b) the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 5% lower than the level of the second 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 also less than 0.1.
  • the difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the fructose- 1 ,6-diphosphate in the mesocarp tissue of the fruit of the parental oil palm plant 19 to 21 weeks after pollination thereof is 33% to 82% higher than the level of the fructose- 1,6- diphosphate in the mesocarp tissue of the fruit of the reference oil palm plant 19 to 21 weeks after pollination thereof, and the level of the 3-phosphoglyceric acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 17 weeks after pollination thereof is 17% to 31% lower than the level of the 3-phosphoglyceric acid in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 1 weeks after
  • the difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the fructose- 1 ,6-diphosphate in the mesocarp tissue of the fruit of the parental oil palm plant 19 to 21 weeks after pollination thereof is 33% to 82% higher than the level of the fructose- ,6- diphosphate in the mesocarp tissue of the fruit of the reference oil palm plant 19 to 21 weeks after pollination thereof, and the level of the ribulose-5-phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 6.5% to 32% lower than the level of the ribulose-5 -phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 13
  • the difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the glucose- 1 -phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 19 to 21 weeks after pollination thereof is 30% to 75% higher than the level of the glucose- 1- phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 19 to 21 weeks after pollination thereof, and the level of the 3-phosphoglyceric acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 17 weeks after pollination thereof is 17% to 31% lower than the level of the 3-phosphoglyceric 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 first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the glucose- 1 -phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 19 to 21 weeks after pollination thereof is 30% to 75% higher than the level of the glucose- 1- phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 19 to 21 weeks after pollination thereof, and the level of the ribulose-5-phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 6.5% to 32% lower than the level of the ribulose-5-phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 5 weeks after pollination thereof.
  • the difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the glycerol- 3-phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 1 1 to 19 weeks after pollination thereof is 22% to 190% higher than the level of the glycerol-3- phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 19 weeks after pollination thereof, and the level of the 3-phosphoglyceric acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 17 weeks after pollination thereof is 17% to 31% lower than the level of the 3-phosphoglyceric 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 first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the glycerol- 3-phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 19 weeks after pollination thereof is 22% to 190% higher than the level of the g ycerol-3- phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 11 to 19 weeks after pollination thereof, and the level of the ribulose-5 -phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 6.5% to 32% lower than the level of the ribulose- -phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks after poll
  • the difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the pyruvic acid in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 13 weeks after pollination thereof is 33% to 82% higher than the level of the pyruvic acid in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 13 weeks after pollination thereof, and the level of the 3-phosphoglyceric acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 17 weeks after pollination thereof is 17% to 31% lower than the level of the 3-phosphoglyceric 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 first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the pyruvic acid in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 13 weeks after pollination thereof is 33% to 82% higher than the level of the pyruvic acid in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 13 weeks after pollination thereof, and the level of the ribulose-5 -phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 6.5% to 32% lower than the level of the ribulose-5 -phosphate 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 first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the ribose-5- phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 11 to 15 weeks after pollination thereof is 47% to 75% higher than the level of the ribose-5-phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 15 weeks after pollination thereof, and the level of the 3-phosphoglyceric acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 17 weeks after pollination thereof is 17% to 31% lower than the level of the 3-phosphoglyceric 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 first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the ribose-5- phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 1 1 to 15 weeks after pollination thereof is 47% to 75% higher than the level of the ribose-5 -phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 1 1 to 15 weeks after pollination thereof, and the level of the ribulose-5 -phosphate in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 15 weeks after pollination thereof is 6.5% to 32% lower than the level of the ribulose-5 -phosphate in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 15 weeks
  • the level of the first metabolite i.e. a metabolite selected from the group consisting of fructose- 1 ,6-diphosphate, glucose- 1 - phosphate, gIycerol-3-phosphate, pyruvic acid, and ribose-5-phosphate
  • the level of the second metabolite i.e.
  • a metabolite selected from the group consisting of 3- phosphoglyceric acid and ribulose-5 -phosphate, in the mesocarp tissue of the fruit of the parental oil palm plant and the levels of the first and second metabolites 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. 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 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 each of the first and second metabolites in the mesocarp tissue o the fruit of the parental oil palm plant and the levels of the first and second metabolites in mesocarp tissue of a fruit of a reference oil palm plant to obtain the high-yielding oil palm plant.
  • 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 first metabolite and the level of a second metabolite in mesocarp tissue of a fruit of a test oil palm plant 11 to 21 weeks after pollination thereof; (ii) determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of a reference oil palm plant, also 11 to 21 weeks after pollination thereof, wherein the difference is that the level of the first metabolite in the mesocarp tissue of the fruit of the test oil palm plant is at least 15% higher than the level of the first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and the level of the second metabolite in
  • metabolites noted above as being useful in the other method for obtaining a high-yielding oil palm plant i.e. a first metabolite selected from the group consisting of fructose- 1 ,6-diphosphate, glucose- 1 -phosphate, glycerol-3-phosphate, pyruvic acid, and ribose- 5 -phosphate, and a second metabolite selected from the group consisting of 3- phosphoglyceric acid and ribulose-5-phosphate, are also useful in the method for predicting oil yield of a test oil palm plant.
  • the step of determining the levels of the first and second metabolites in mesocarp tissue of a fruit of a test oil palm plant 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 each of the first and second 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 spectrometry,
  • the step of determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second 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 metabolites.
  • 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.
  • 1 1 to 21 weeks after pollination thereof for example 1 1-19 weeks, 1 1-17 weeks, 1 1 -15 weeks, 1 1-13 weeks, 13-21 weeks, 13-19 weeks, 13-17 weeks, 13-15 weeks, 15-21 weeks, 15-19 weeks, 15-17 weeks, 17-21 weeks, 17-19 weeks, 19-21 weeks, 1 1 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, or 21 weeks after pollination thereof.
  • the levels of the first and second metabolites in the mesocarp tissue of the fruit of the parental oil palm plant may be determined as mean levels.
  • the levels of the first and second metabolites in mesocarp tissue of a fruit of a reference oil palm plant may be determined before, during, or after determination of the levels of the first and second metabolites in mesocarp tissue of the fruit of the parental oil palm plant.
  • the difference is that (a) the level of the first metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 15% higher than the level of the first 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 , and (b) the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant is at least 5% lower than the level of the second 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 also 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
  • the difference between the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the parental oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant is that the level of the fructose- 1 ,6-diphosphate in the mesocarp tissue of the fruit of the parental oil palm plant 19 to 21 weeks after pollination thereof is 33% to 82% higher than the level of the fructose- 1 ,6-diphosphate in the mesocarp tissue of the fruit of the reference oil palm plant 19 to 21 weeks after pollination thereof, and the level of the 3-phosphoglyceric acid in the mesocarp tissue of the fruit of the parental oil palm plant 13 to 17 weeks after pollination thereof is 17% to 31% lower than the level of the 3-phosphoglyceric acid in the mesocarp tissue of the fruit of the reference oil palm plant 13 to 17 weeks after
  • the predicting step may be carried out, for example, based on the amount of the difference in the level of the first metabolite and the level of the second metabolite in the mesocarp tissue of the fruit of the test oil palm plant and the level of the first metabolite and the level of the second metabolite in mesocarp tissue of the fruit of the reference oil palm plant, and/or based on correlations between levels of the metabolites 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 each of the first and second metabolites.
  • kits for obtaining a high-yielding oil palm plant or predicting oil yield of a test oil palm plant comprises: (i) a first reagent for quantitative detection of a first metabolite; (ii) a second reagent for quantitative detection of a second metabolite; (iii) an extract of a mesocarp tissue of a fruit of a reference oil palm plant 11 to 21 weeks after pollination thereof; and (iv) instructions indicating use of the first reagent and the second reagent for determining whether there is a difference between the level of the first metabolite and the level of the second metabolite in mesocarp tissue of a fruit of the parental or test oil palm plant, also 11 to 21 weeks after pollination thereof, and the level of the first metabolite and the level of the second 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 first reagent for quantitative detection of a first metabol
  • the metabolites described above as being useful in the other method for obtaining a high- yielding oil palm plant i.e. a first metabolite selected from the group consisting of fructose- 1 ,6-diphosphate, glucose- 1 -phosphate, glycerol-3 -phosphate, pyruvic acid, and ribose-5-phosphate, and a second metabolite selected from the group consisting of 3- phosphoglyceric acid and ribulose-5-phosphate, are also useful in the kit for obtaining a high-yielding oil palm plant.
  • the first reagent for quantitative detection of the first metabolite and the second reagent for quantitative detection of the second 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
  • 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.
  • Collection of oil palm mesocarp tissue Fruits were harvested from plants of the high-yielding and low-yielding screening populations at week 16 post-pollination.
  • LC-MS liquid chromatography-mass spectrometry
  • 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 I 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 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. Samples were analyzed using an Agilent CE-TOFMS system (Agilent
  • 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.
  • 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 glycero 1-3 -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.
  • palm oil contents of mesocarp tissue of fruits of oil palm plants can be enhanced by use of these identified metabolites as metabolite-based markers to obtain high-yielding oil palm plants and to predict oil yield of test oil palm plants, as described herein.
  • Example 2
  • 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 ⁇ - was injected into a programmable injector. The total run time is 55 minutes.
  • FIGS. 4-13 Results are shown in FIGS. 4-13.
  • FIGS. 4 and 5 provide heat map 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 1 1 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 11 to 21 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- ⁇ or dimethyl sulfoxide- ⁇ -
  • the level of a first metabolite and the level of a second metabolite in the samples are then determined.
  • the first metabolite is selected from the group consisting of malic acid and pyruvic acid
  • the second metabolite is selected from the group consisting of 2-oxoglutaric acid, isocitric acid, and citramalic acid.
  • the first metabolite is selected from the group consisting of fructose- 1 ,6-diphosphate, glucose- 1 -phosphate, glycerol-3-phosphate, pyruvic acid, and ribose-5-phosphate
  • the second metabolite is selected from the group consisting of 3-phosphogIyceric acid and ribulose-5-phosphate.
  • the level of malic acid and the level of isocitric acid in the sample can be determined.
  • the difference can be that the level of the first metabolite in the sample is at least 15% higher than the level of the first metabolite in the mesocarp tissue of the fruit of the reference oil palm plant, and that the level of the second metabolite in the sample is at least 5% lower than the level of the second metabolite in the mesocarp tissue of the fruit of the reference oil palm plant.
  • the difference can be that the level of malic acid is at least 15% higher in the sample in comparison to the level of malic acid in the mesocarp tissue of the fruit of the reference oil palm plant, and that the level of isocitric acid is at least 5% lower in the sample in comparison to the level of isocitric acid 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 malic acid is at least 15% higher in the sample in comparison to the level of malic acid in the mesocarp tissue of the fruit of the reference oil palm plant and for which the level of isocitric acid is at least 5% lower in the sample in comparison to the level of isocitric acid 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 of a first metabolite and one or more of a second metabolite are then determined in the mesocarp tissue samples obtained from the progeny of both crosses, and compared with respect to the concentrations of the first and second 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 malic acid and lower levels of isocitric acid
  • 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, NM analysis, or an enzyme-based biosensor, 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.
  • 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 of mesocarp samples:
  • 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.
  • each of the first and second metabolites noted in Example 3 can be used 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
  • 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, or an enzyme-based biosensor.
  • 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 malic acid in the mesocarp tissue of the fruit of the parental or test oil palm plant being at least 15% higher, e.g. between 43% and 100% higher, than the level of malic acid in the mesocarp tissue of the
  • the level of isocitric acid in the mesocarp tissue of the fruit of the parental or test oil palm plant being at least 5% lower, e.g. between 13% and 35% lower, than the level of isocitric acid in the mesocarp tissue of the fruit of the reference oil palm plant, 1 1 to 21 weeks, e.g. 11 to 19 weeks, after pollination thereof.
  • Malic acid and isocitric acid also can be detected and quantified by simple HPLC analysis and spectrophotometric detection, e.g. using a column of strong cation exchange resin, eluting with dilute mineral acid, and monitoring of the column effluent by UV spectroscopy at 210 nm, as taught for example by Turkelson et al., 50 Analytical Chemistry 1420 (1978).
  • NMR analysis can also be used to efficiently compare malic acid and isocitric acid in biological samples, e.g.
  • Malic acid also can be detected and quantified by use of an enzyme-based biosensor, e.g. an amperometric biosensor based on malate quinone oxidoreductase, with phenazine methosulfate as an electrochemical mediator, as taught for example by Bucur et al., 21 Biosensors & Bioelectronics 2290-2297 (2006).
  • an enzyme-based biosensor e.g. an amperometric biosensor based on malate quinone oxidoreductase, with phenazine methosulfate as an electrochemical mediator, as taught for example by Bucur et al., 21 Biosensors & Bioelectronics 2290-2297 (2006).
  • the combination comparisons of malic acid and isocitric acid can be carried out based on these methods by direct comparison of extract profiles at a consistent concentration.
  • 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.
  • Glycero 1-3 -phosphate Adenosine triphosphate (“ATP") TABLE 2.
  • Metabolites useful for identifying high-yielding 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.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Botany (AREA)
  • Developmental Biology & Embryology (AREA)
  • Environmental Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Physiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne des procédés d'obtention d'un palmier à huile avec un rendement élevé, qui comprend les étapes consistant à déterminer les taux des premier et second métabolites dans des tissus mésocarpes d'un fruit du palmier à huile mère, 11 à 21 semaines après sa pollinisation, déterminer s'il y a une différence entre ces taux et les taux des premier et second métabolites dans un tissu mésocarpe d'un fruit d'un palmier à huile de référence, également 11 à 21 semaines après sa pollinisation, la différence étant que le taux du premier métabolite est au moins 15 % plus élevé, et le taux du second métabolite est au moins 5 % moins élevé, pour la plante mère par rapport à la plante de référence, et sélectionner la descendance de la plante mère sur la base de la différence pour obtenir la plante avec un rendement élevé. L'invention concerne également des procédés permettant de prédire le rendement en huile d'un palmier à huile à tester et des kits pour l'obtention d'un palmier à huile avec un rendement élevé.
PCT/MY2012/000242 2011-12-14 2012-08-28 Procédés d'obtention de palmiers à huile avec un rendement élevé WO2013089558A1 (fr)

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SG11201403226SA SG11201403226SA (en) 2011-12-14 2012-08-28 Methods for obtaining high-yielding oil palm plants
CR20140281A CR20140281A (es) 2011-12-14 2014-06-13 Métodos para la obtención de plantas de alto rendimiento de palma de aceite

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WO2015034345A1 (fr) 2013-09-04 2015-03-12 Sime Darby Malaysia Berhad Procédés de prédiction du rendement en huile d'une installation d'essai de fabrication d'huile de palme
WO2015034344A1 (fr) * 2013-09-04 2015-03-12 Sime Darby Malaysia Berhad Procédés de prédiction du rendement en huile d'un palmier à huile testé
CN112034070A (zh) * 2020-09-09 2020-12-04 福建省沙县水南国有林场(福建沙县罗岩山森林公园工作站) 一种油茶高产与低产品种组代谢表达谱标记识别方法

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015034345A1 (fr) 2013-09-04 2015-03-12 Sime Darby Malaysia Berhad Procédés de prédiction du rendement en huile d'une installation d'essai de fabrication d'huile de palme
WO2015034344A1 (fr) * 2013-09-04 2015-03-12 Sime Darby Malaysia Berhad Procédés de prédiction du rendement en huile d'un palmier à huile testé
CN112034070A (zh) * 2020-09-09 2020-12-04 福建省沙县水南国有林场(福建沙县罗岩山森林公园工作站) 一种油茶高产与低产品种组代谢表达谱标记识别方法

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