# US 11,307,139 B2 · Non-destructive assay for soybean seeds using near infrared analysis

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- Number: US 11,307,139 B2
- Document ID: US11307139B2
- Status: Granted
- Kind: B2
- Application number: 16/490,261
- Filed: 2018-02-26
- Published: 2019-12-19
- Granted: 2022-04-19
- Earliest claimed: 2017-03-03
- Assignees: Pioneer Hi-Bred International, Inc.
- Applicants: Pioneer Hi-Bred International, Inc.
- Inventors: Ross M Allen; John D Everard; Min Ren
- Web page: https://patentgrep.com/patent/US11307139B2

## CPC

- G01N 21/359: using near infrared light
- B07C 5/3425: {of granular material, e.g. ore particles, grain (investigation moving fluids or granular solids using optical means G01N21/85)}
- C12N 15/8245: {involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis}
- C12N 15/8247: {involving modified lipid metabolism, e.g. seed oil composition}
- G01N 21/3563: for analysing solids; Preparation of samples therefor
- G01N 21/84: Systems specially adapted for particular applications
- G01N 33/025: {Fruits or vegetables}
- G01N 33/10: Starch-containing substances, e.g. dough
- G01N 2021/3595: {using FTIR}
- G01N 2021/8466: {Investigation of vegetal material, e.g. leaves, plants, fruits}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- B07C: POSTAL SORTING; SORTING INDIVIDUAL ARTICLES, OR BULK MATERIAL FIT TO BE SORTED PIECE-MEAL, e.g. BY PICKING (specially adapted for a specific purpose covered by another class, see the relevant place, e.g. A43D33/06, B23Q7/12)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Disclosed are methods and systems for spectral imaging of soybean samples to accurately and non-destructively measure the amount of sucrosyl-oligosaccharide in the soybean samples. Populations containing modified and unmodified soybean seeds and having varying amounts of sucrosyl-oligosaccharides, oil or protein can be sorted and separated and further used in soybean processing or breeding.

## Claims

### Claim 1

A method for measuring the amount of a sucrosyl-oligosaccharide-in a soybean seed, the method comprising:
(a) directing near infrared light from a light source onto a soybean seed to form modified light from the soybean seed;
(b) receiving the modified light in an imaging device;
(c) measuring the amount of a sucrosyl-oligosaccharide in the soybean seed based on the received modified light, the amount of the sucrosyl-oligosaccharide being measured to an accuracy that is within 1.5 wt. % of the amount measured using a standard reference analytical method; and
(d) transporting the seed to a first location when the amount of sucrosyl-oligosaccharide measured is below a threshold value between 1 wt. % and 0.1 wt. % and transporting the seed to a different second location when the amount of sucrosyl-oligosaccharide measured is at or above the threshold value.

### Claim 2 (depends on 1)

The method of claim 1, wherein the threshold value is between 0.5 wt. % and 0.1 wt.-%.

### Claim 3 (depends on 1)

The method of claim 1, wherein a plurality of seeds are measured together in step a, and wherein the amount of a sucrosyl-oligosaccharide measured in step (c) is an average amount for the plurality of seeds.

### Claim 4 (depends on 3)

The method of claim 3, wherein the plurality of seeds comprises at least 10 and less than 100 seeds.

### Claim 5 (depends on 3)

The method of claim 3, wherein the plurality of seeds comprises at least 1 kg of seeds and less than 1,000 kg of seeds.

### Claim 6 (depends on 1)

The method of claim 1, wherein an individual seed is measured in step (a).

### Claim 7 (depends on 6)

The method of claim 6, wherein the method is an automated method, and further comprising separating the individual seed from a plurality of seeds prior to step (a).

### Claim 8 (depends on 1)

The method of claim 1, wherein the seed is genetically modified to overexpress a diglyceride acyltransferase.

### Claim 9 (depends on 1)

The method of claim 1, wherein the sucrosyl-oligosaccharide measured is stachyose.

### Claim 10 (depends on 9)

The method of claim 9 wherein the imaging device in step (c) is calibrated using a plurality of soybean seeds having variable stachyose contents falling in a range that includes values of less than 0.3 wt. % stachyose and more than 4.5 wt. % stachyose.

### Claim 11 (depends on 1)

The method of claim 1, wherein the modified light comprises transmitted light.

### Claim 12 (depends on 1)

The method of claim 1, wherein the modified light comprises reflected light.

### Claim 13

A method for measuring stachyose in a population of soybean seeds, the method comprising
(a) directing near infrared light from a light source onto a first and second subsample of the population of soybean seeds to form a first and second modified light from the soybean seeds;
(b) receiving the first and second modified light in an imaging device;
(c) measuring the amount of stachyose in the first and second subsamples based on the received first and second modified light, wherein (i) the first and second subsamples are separated when the amount of stachyose measured differs by at least 1 percentage point between the first and second subsamples and (ii) wherein the first and second subsamples are combined when the amount of stachyose differs by less than 0.2 percentage points.

### Claim 14 (depends on 13)

The method of claim 13, wherein the population comprises genetically modified and unmodified soybean seeds.

### Claim 15 (depends on 14)

The method of claim 14, wherein the modified seeds comprise a modified diacylglycerol transferase.

### Claim 16

A method for processing soybean seeds genetically modified to contain high oil, high protein, modified amino acid content, or a combination thereof compared with unmodified soybean seeds in a plurality of seeds comprising the modified soybean seeds and the unmodified soybean seeds, the method comprising
(a) directing near infrared light from a light source onto a sample comprising a soybean seed to form modified light from the soybean seed;
(b) receiving the modified light in an imaging device;
(c) measuring the amount of a sucrosyl-oligosaccharide in the sample based on the received modified light, the amount of sucrosyl-oligosaccharide being measured to an accuracy that is within 1 wt. % of the amount measured using a standard reference analytical method; and
(d) repeating steps (a) to (c) for at least 10 samples, wherein an amount of sucrosyl-oligosaccharide below a threshold value indicates high oil, high protein, modified amino acid content, or a combination thereof in the soybean seed; and
(e) separating soybean seeds above the threshold value from soybean seeds below the threshold value, wherein at least 90% of the soybean seeds below the threshold value are the modified soybean seeds or wherein at least 90% of the seeds above the threshold value are the unmodified soybean seeds.

### Claim 17 (depends on 16)

The method of claim 16, wherein at least one of the modified seeds separated in step (e) is grown and crossed with the same or a different soybean plant to produce progeny seed.

### Claim 18 (depends on 16)

The method of claim 16, wherein the method is an automated method, and further comprising separating the sample comprising the soybean seed from the plurality of seeds prior to step (a).

### Claim 19 (depends on 16)

The method of claim 16, further comprising measuring the amount of oil in the seed based on the received modified light and wherein at least 90% of the soybean seeds above the threshold value are the modified soybean seeds or wherein at least 90% of the seeds below the threshold value are the unmodified soybean seeds.

### Claim 20 (depends on 16)

The method of claim 16, wherein the sucrosyl-oligosaccharide is stachyose.