Genetic markers for Myb28

Patent metadata

Field	Value
Publication number	US-10472685-B2
Application number	US-201615359471-A
Country	US
Kind code	B2
Filing date	Nov 22, 2016
Priority date	Sep 13, 2012
Publication date	Nov 12, 2019
Grant date	Nov 12, 2019

Abstract

The present invention relates to a method for determining the genotype of a Cruciferous vegetable plant for a plant with an increased glucosinolate level, comprising obtaining a sample of nucleic acids from said plant or a portion thereof and detecting in said nucleic acids a polymorphism at the Myb28 locus that is genetically linked to an increased glucosinolate level. The polymorphism may comprises at least one of: a) a single nucleotide polymorphism (SNP) at a position corresponding to nucleotide 83, 136, 226, 563, 610, 830, 995, 1116, 1513, 1577, 1606, 1620, 1825, 1863, 1877 or 2026 of SEQ ID NO: 1, or b) a polymorphism in the number of nucleotides present between nucleotides 323 and 332, between nucleotides 521 and 524, between nucleotides 783 and 786, between nucleotides and 909 and 914, between nucleotides 1365 and 1369, between 1811 and 1821, or between nucleotides 2046 and 2056 of SEQ ID NO: 1, or c) a polymorphism in the number of nucleotides present between nucleotides 836 and 837, between nucleotides 867 and 868, or between nucleotides 943 and 944 of SEQ ID NO: 1.

First claim

The invention claimed is:

1. A method for identifying a Cruciferous vegetable plant with an increased glucosinolate level, said method comprising:
(a) obtaining a sample of nucleic acids from said plant or a portion thereof;
(b) detecting in said nucleic acids a locus comprising a polymorphism, wherein said locus is indicative of said increased glucosinolate level, and wherein said polymorphism comprises a SNP at position 1116 of SEQ ID NO:1;
(c) crossing a Cruciferous vegetable plant comprising said polymorphism with a second Cruciferous vegetable plant;
(d) obtaining seeds from said cross; and
(e) growing at least one Cruciferous vegetable plant from said seeds, wherein said Cruciferous vegetable plant grown from said seeds comprises said polymorphism and has an increased glucosinolate level when compared with a Cruciferous vegetable plant of the same variety which does not comprise said polymorphism.

2. The method according to claim 1 , wherein the step of detecting comprises PCR or DNA hybridization.

3. The method according to claim 1 , wherein determining the genotype comprises a co-dominant assay.

4. The method according to claim 1 , wherein the method further comprises assaying the phenotype of said Cruciferous vegetable plant comprising said polymorphism for an increased level of glucosinolate.

5. The method according to claim 1 , wherein the glucosinolate is 4-methylsulphinylbutyl glucosinolate (MSB), 3-methylsulphinylpropyl glucosinolate (MSP) or combinations thereof.

6. The method according to claim 1 , wherein the Cruciferous vegetable plant comprises at least one glucosinolate in amount of at least 10 micromol/g dry weight.

7. The method according to claim 1 , wherein the Cruciferous vegetable plant comprises 4-methylsulphinylbutyl glucosinolate (MS B), 3-methylsulphinylpropyl glucosinolate (MSP) or combinations thereof in an amount of at least 10 micromol/g dry weight.

8. The method according to claim 1 , wherein the Cruciferous vegetable plant is broccoli.

9. The method of claim 1 , which method further comprises the step of storing the result of the step of detecting the polymorphism on a computer readable medium.

10. The method according to claim 1 , wherein said plant was produced by crossing a Cruciferous vegetable plant having increased glucosinolate levels with a second Cruciferous vegetable plant lacking increased glucosinolate levels.

11. The method according to claim 1 , further comprising crossing the Cruciferous vegetable plant with an increased glucosinolate level with itself or a third plant to produce a progeny plant of a subsequent generation.

12. The method according to claim 11 , further comprising the steps of: (a) crossing the progeny plant of a subsequent generation with itself or a second plant; and (b) repeating step (a) for an additional 3-10 generations with sufficient inbreeding to produce an inbred Cruciferous vegetable plant comprising an increased level of glucosinolate, wherein the progeny plant of at least one subsequent generation is screened for the presence of a polymorphism at the Mvb28 locus genetically linked to glucosinolate production.

13. The method according to claim 12 , wherein said progeny plant of a subsequent generation is selected for crossing based on the presence of glucosinolates and a desired trait.

14. The method according to claim 13 , wherein the progeny plant of a. subsequent generation is selected at each generation for crossing based on the presence of an increased glucosinolate level and the desired trait.

15. The method of claim 1 , wherein said polymorphism further comprises a SNP at position 83, 136, 226, 563, 610, 830, 995, 1513, 1577, 1606, 1620, 1825, 1863, 1877 or 2026 of SEQ ID NO: 1.

Assignees

• Seminis Vegetable Seeds Inc
• Plant Bioscience Ltd

Inventors

• Mithen Richard F
• Traka Maria
• Brugmans Bart W

Classifications

• C12Q2600/156

  Polymorphic or mutational markers · CPC title

• C12N15/8243

  involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine · CPC title

• C12Q2600/13

  Plant traits · CPC title

• C12Q1/6895 Primary

  for plants, fungi or algae · CPC title

• C12Q2600/158

  Expression markers · CPC title