High throughput screening of fatty acid composition

Abstract

A method for the high throughput screening of fatty acid characteristics in seeds is provided. The method comprises feeding seeds individually to a sampling station; removing a sample from the seed in the sampling station; conveying the sample to a compartment in a sample tray; converting extracted oil from the sample in the sample tray to form a mixture of fatty acid methyl esters; and analyzing the mixture of fatty acid methyl esters from the sample to determine the fatty acid profile of the corresponding seed.

1 . An automated method for determining the fatty acid composition of a plurality of seeds, the method comprising: 
 sequentially feeding a seed to a sampling station;    holding the seed in a sampling station;    scraping a sample from the seed being held in the sampling station;    conveying the sample to an individual compartment in a sample tray;    extracting oil from the sample in the sample tray;    converting extracted oil from the sample in the sample tray to form a mixture of fatty acid esters; and    analyzing the mixture of fatty acid esters from the sample to determine the fatty acid profile of the corresponding seed.    
   
   
       2 . The method of  claim 1  wherein the steps of extracting oil and converting extracted oil comprise contacting the sample in the sample tray with a solvent to form the mixture of fatty acid esters.  
   
   
       3 . The method of  claim 2 , wherein the solvent is selected from the group consisting of hexane, benzene, tetrahydrofuran, dimethyl sulfoxide, trimethylsulfonium hydroxide, petroleum ether, methylene chloride, and toluene.  
   
   
       4 . The method of  claim 2 , wherein the solvent is toluene.  
   
   
       5 . The method of  claim 1 , wherein the step of analyzing comprises separating and detecting the mixture of fatty acid esters using gas chromatography.  
   
   
       6 . The method of  claim 1 , wherein the sample is scraped from the seed without affecting the germination viability of the seed.  
   
   
       7 . The method of  claim 1 , wherein the sample tray comprises a 96-well microtiter plate.  
   
   
       8 . The method of  claim 1 , wherein the sample tray comprises a 384-well microtiter plate.  
   
   
       9 . A method for high throughput screening of oil seeds, the method comprising: 
 providing tissue samples from a plurality of oil seeds in individual compartments of a sample tray;    contacting each tissue sample in the sample tray with toluene to produce a mixture comprising fatty acid methyl esters;    analyzing the mixture of fatty acid methyl esters from each sample to determine the fatty acid profile of the corresponding oil seed; and    selecting seed based on the presence or absence of a desired fatty acid characteristic.    
   
   
       10 . The method of  claim 9 , wherein the step of analyzing comprises separating and detecting the fatty acid methyl esters using gas chromatography.  
   
   
       11 . The method of  claim 9 , wherein the fatty acid profile of the corresponding oil seed is determined in less than about 10 minutes from the time in which an individual tissue sample is contacted with toluene.  
   
   
       12 . The method of  claim 9 , wherein the method further comprises collecting the tissue samples without affecting the germination viability of the seed.  
   
   
       13 . The method of  claim 9 , wherein the sample tray comprises at least 96 individual compartments.  
   
   
       14 . The method of  claim 9 , wherein the oil seeds are selected from the group consisting of soybean, corn, canola, rapeseed, sunflower, peanut, safflower, palm and cotton.  
   
   
       15 . The method of  claim 14 , wherein the oil seed is rapeseed and the desired fatty acid characteristic is an erucic acid content of greater than about 2%.  
   
   
       16 . The method of  claim 14 , wherein the seed is rapeseed and the desired fatty acid characteristic is an erucic acid content of greater than about 45%.  
   
   
       17 . The method of  claim 14 , wherein the seed is rapeseed and the desired fatty acid characteristic is a linolenic acid content of less than about 3.5%.  
   
   
       18 . The method of  claim 14 , wherein the seed is rapeseed and the desired fatty acid characteristic is an oleic acid content of greater than about 70%.  
   
   
       19 . The method of  claim 14 , wherein the seed is rapeseed and the desired fatty acid characteristic is a saturated fat content of less than about 7%.  
   
   
       20 . The method of  claim 14 , wherein the seed is rapeseed and the desired fatty acid characteristic is a saturated fat content of less than about 6%.  
   
   
       21 . The method of  claim 14 , wherein the seed is rapeseed and the desired fatty acid characteristic is a saturated fat content of less than about 5%.  
   
   
       22 . The method of  claim 14 , wherein the seed is rapeseed and the desired fatty acid characteristic is an oleic acid content of greater than about 70% and a linolenic acid content of less than about 3.5%.  
   
   
       23 . The method of  claim 14 , wherein the seed is rapeseed and the desired fatty acid characteristic is an oleic acid content of greater than about 70%, a linolenic acid content of less than about 3.5%, and a saturated fat content of less than about 7%.  
   
   
       24 . The method of  claim 14 , wherein the seed is rapeseed and the desired fatty acid characteristic is an erucic acid content of greater than about 45% and a linolenic acid content of less than about 3.5%.  
   
   
       25 . The method of  claim 14 , wherein the seed is sunflower and the desired fatty acid characteristic is an oleic acid content of from about 40% to about 70%.  
   
   
       26 . The method of  claim 14 , wherein the seed is sunflower and the desired fatty acid characteristic is an oleic acid content of greater than about 70%.  
   
   
       27 . The method of  claim 14 , wherein the seed is sunflower and the desired fatty acid characteristic is a stearic acid content of greater than about 6%.  
   
   
       28 . The method of  claim 14 , wherein the seed is sunflower and the desired fatty acid characteristic is a saturated fat content of less than about 8%.  
   
   
       29 . The method of  claim 14 , wherein the seed is sunflower and the desired fatty acid characteristic is an oleic acid content of greater than about 70% and a saturated fat content of less than about 8%.  
   
   
       30 . The method of  claim 14 , wherein the seed is sunflower and the desired fatty acid characteristic is an oleic acid content of greater than about 70%, a stearic acid content of greater than about 6%, and a saturated fat content of less than about 8%.  
   
   
       31 . The method of  claim 14 , wherein the seed is soybean and the desired fatty acid characteristic is a linolenic acid content of less than about 8%.  
   
   
       32 . The method of  claim 9 , wherein the method further comprises cultivating plants from the selected seed; and harvesting seeds from the cultivated plants.  
   
   
       33 . A method of bulking up a quantity of seed having a desired fatty acid characteristic, the method comprising: 
 (a) removing a sample from each seed in a population without affecting the germination viability of the seeds;    (b) contacting each sample with a solvent to form a mixture comprising fatty acid methyl esters;    (c) analyzing the mixture of fatty acid methyl esters from each sample to determine the fatty acid profile of the corresponding seed;    (d) selecting seeds having at least one desired fatty acid characteristic;    (e) cultivating plants from the selected seeds;    (f) recovering seed from the cultivated plants; and    repeating steps (a) through (f) for one or more generations.    
   
   
       34 . The method of  claim 33 , wherein the solvent is selected from the group consisting of hexane, benzene, tetrahydrofuran, dimethyl sulfoxide, trimethylsulfonium hydroxide, petroleum ether, methylene chloride, and toluene.  
   
   
       35 . The method of  claim 34 , wherein the solvent is toluene.  
   
   
       36 . The method of  claim 33 , wherein the step of analyzing comprises separating and detecting the mixture of fatty acid esters using gas chromatography.  
   
   
       37 . The method of  claim 33 , wherein the fatty acid profile of the corresponding oil seed is determined in less than about 10 minutes from the time in which an individual tissue sample is contacted with solvent.  
   
   
       38 . The method of  claim 33 , wherein the seeds are oil seeds selected from the group consisting of soybean, corn, canola, rapeseed, sunflower, peanut, safflower, palm and cotton.