Methods for genotyping mature cotton fibers and textiles

• US 9,290,819 B2
• Filed: 12/29/2014
• Issued: 03/22/2016
• Est. Priority Date: 11/12/2008
• Status: Active Grant

• Current Assignee: APDN (B.V.I.) Inc. (Applied DNA Sciences Incorporated)
• Original Assignee: APDN (B.V.I.) Inc. (Applied DNA Sciences Incorporated)
• Inventors: Liang, MingHwa Benjamin, Shu Kin So, Stephane
• Primary Examiner(s): Joike, Michele K
• Assistant Examiner(s): Brown, Mindy G
• Application Number:
  US14/584,309
• Publication Number:
  US 20150191799A1
• Time in Patent Office: 449 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C12Q 1/6895 for plants, fungi or algae
• CPC Class Codes: C12Q 2600/13 Plant traits
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers

Abstract

Methods for distinguishing between cotton cultivars of a specific species by analyzing a sample of mature cotton fibers from raw cotton materials or from textile goods are disclosed. DNA is extracted from the mature cotton fiber sample and subjected to PCR techniques which enable the identification of the cultivar of a particular cotton species utilized in the textile or cotton material of interest.

Claims

1. A method for identifying a cotton cultivar comprising:

collecting one or more mature cotton fibers from a manufactured textile;

extracting genomic DNA from the one or more mature cotton fibers collected;

amplifying at least a portion of the extracted genomic DNA with a PCR-based technique using one or more primer sets and thereby producing one or more amplicons;

generating a variable amplicon length polymorphism profile of the extracted genomic DNA;

comparing the variable length polymorphism profile with a known variable amplicon length polymorphism profile; and

thereby identifying at least one cotton cultivar from the manufactured textile.

2. The method of claim 1, wherein at least one cotton cultivar identified is from the species G. barbadense.

3. The method of claim 2, wherein at least one cotton cultivar identified is from the species G. hirsutum.

4. The method of claim 1, wherein the amplifying comprises:

using two or more sets of specific primers which target length polymorphisms between a first cotton cultivar and a second cotton cultivar.

5. The method of claim 2, wherein the amplifying comprises:

using at least one set of specific primers targeting sequences differing in length between each of at least three G. barbadense cultivars.

6. The method of claim 4, wherein polymorphic sequences of the cotton cultivars targeted by the primers differ in another detectable physical characteristic in addition to the difference in sequence lengths.

7. The method of claim 6, wherein the additional detectable physical characteristic comprises a difference in a nucleotide composition.

8. The method of claim 1, wherein the genomic DNA extracted from the one or more mature cotton fibers comprises chloroplast DNA.

9. The method of claim 1, wherein the genomic DNA extracted from the one or more mature cotton fibers comprises mitochondrial DNA.

10. The method of claim 1, wherein the genomic DNA extracted from the one or more mature cotton fibers comprises nuclear DNA.

11. The method of claim 1, further comprising detecting a size of at least one of the amplicons by gel electrophoresis.

12. The method of claim 1, further comprising detecting a size of at least one of the amplicons by capillary electrophoresis.

13. The method of claim 1, further comprising detecting a size of at least one of the amplicons with real time PCR.

14. The method of claim 1, wherein the at least one cotton cultivar is from a cotton species selected from the group consisting of G. barbadense, G. hirsutum, G. arboreum, or G. herbaceum.

25. The method of claim 1, wherein the manufactured textile comprises raw cotton, ginned cotton, yarn blended with cotton and or a fabric blended with cotton.

26. The method of claim 1, wherein the amplifying of said genomic DNA comprises using a nested set of primers.

32. The method of claim 13, further comprising quantifying one or more of the amplified products by real-time PCR.

33. The method of claim 4, wherein the first cotton cultivar and the second cotton cultivar are from a cotton species selected from the group consisting of G. barbadense, G. hirsutum, G. arboreum, or G. herbaceum.

15. A method for verifying the authenticity of a manufactured textile comprising:

collecting mature cotton fibers from the manufactured textile;

extracting genomic DNA from the mature cotton fibers collected;

amplifying at least a portion of the extracted genomic DNA with a PCR-based technique using one or more primer sets producing one or more amplified products forming a variable amplicon length polymorphism profile for the mature cotton fibers from the manufactured textile;

comparing the variable amplicon length polymorphism profile from the mature cotton fibers with one or more known variable amplicon length polymorphism profiles;

identifying the mature cotton fibers from the manufactured textile as belonging to one or more cotton cultivars; and

thereby determining whether the manufactured textile is authentic or counterfeit.

16. The method of claim 15, wherein at least one cotton cultivar identified is from the species G. barbadense.

17. The method of claim 15, wherein at least one cotton cultivar identified is from the species G. hirsutum.

18. The method of claim 15, wherein the amplifying further comprises:

using at least one set of specific primers which targets a sequence polymorphism between a first cotton cultivar and a second cotton cultivar.

19. The method of claim 16, wherein the amplifying further comprises:

using two or more sets of specific primers targeting sequence length polymorphisms between G. barbadense cultivars.

20. The method of claim 19, wherein a targeted sequence of a first G. barbadense cultivar and a second G. barbadense cultivar have a difference in a detectable physical characteristic in addition to the sequence length polymorphisms.

21. The method of claim 20, wherein the difference in a detectable physical characteristic of the targeted sequence comprises a difference in a nucleotide composition.

22. The method of claim 15, wherein the genomic DNA extracted from the mature cotton fibers comprises nuclear DNA, chloroplast DNA, and/or mitochondrial DNA.

23. The method of claim 15, wherein the analyzing of the amplified products comprises detecting a size of one or more of the amplified products by gel electrophoresis.

24. The method of claim 15, wherein the analyzing of the amplified products comprises detecting a size of one or more of the amplified products by capillary electrophoresis.

27. The method of claim 15, wherein the amplifying of said genomic DNA comprises using a nested set of primers.

28. The method of claim 27, wherein the cotton cultivar identified is from a cotton species selected from the group consisting of G. barbadense, G. hirsutum, G. arboreum, or G. herbaceum.

29. The method of claim 15, wherein at least one of the cotton cultivars identified is G. arboreum or G. herbaceum.

30. The method of claim 15, wherein the one or more amplified products are produced by real-time PCR.

31. The method of claim 30, further comprising quantifying one or more of the amplified products by real-time PCR.