Methods for manufacturing plant cell walls comprising chitin
View Patent ↗Methods and means are provided for the modification of the reactivity of plant secondary cell walls, particularly in cotton cell walls found in cotton fibers. This can be conveniently achieved by expressing a chimeric gene encoding a Saprolegnia monoica chitin synthase in cotton plants.
1. A plant cell comprising a chimeric gene comprising the following operably linked DNA regions:
a. a plant-expressible promoter,
b. a DNA region coding for a chitin synthase polypeptide, said DNA region comprising:
i. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 95% identical to SEQ ID NO: 2; or
ii. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 95% identical to SEQ ID NO: 4;
c. a transcription termination and polyadenylation region.
2. A plant comprising the at least one plant cell of claim 1 .
3. The plant of claim 2 , wherein said plant is cotton.
4. Fibers comprising a chimeric gene comprising the following operably linked DNA regions:
a. a plant-expressible promoter,
b. a DNA region coding for a chitin synthase polypeptide, said DNA region comprising:
i. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 95% identical to SEQ ID NO: 2; or
ii. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 95% identical to SEQ ID NO: 4;
c. a transcription termination and polyadenylation region obtainable from the plant according to claim 2 .
5. A transgenic seed comprising a chimeric gene comprising the following operably linked DNA regions:
a. a plant-expressible promoter,
b. a DNA region coding for a chitin synthase polypeptide, said DNA region comprising:
i. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 95% identical to SEQ ID NO: 2; or
ii. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 95% identical to SEQ ID NO: 4;
c. a transcription termination and polyadenylation region.
6. A method for the manufacture of a plant cell wall comprising positively charged polysaccharides, said method comprising
a. expressing a chimeric gene comprising the following operably linked DNA regions:
i. a plant-expressible promoter,
ii. a DNA region coding for a chitin synthase polypeptide, said DNA region comprising:
1. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 95% identical to SEQ ID NO: 2; or
2. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 95% identical to SEQ ID NO: 4;
iii. a transcription termination and polyadenylation region,
b. and isolating said plant cell wall.
7. The method according to claim 6 wherein said plant cell wall is a cotton plant cell wall.
8. The method according to claim 7 wherein said cotton plant cell wall is present in a cotton fiber.
9. The plant cell, plant, fiber, or seed according to any one of claims 1 - 4 or 6 , wherein said DNA region comprises:
i. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 98% identical to SEQ ID NO: 2; or
ii. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 98% identical to SEQ ID NO: 4.
10. The plant cell, plant, fiber, or seed according to any one of claims 1 - 4 or 6 , wherein said DNA region comprises:
i. a nucleotide sequence which encodes a chitin synthase polypeptide according to SEQ ID NO: 2; or
ii. a nucleotide sequence which encodes a chitin synthase polypeptide, according to SEQ ID NO: 4.
11. The method according to claim 6 , wherein said DNA region comprises:
i. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 98% identical to SEQ ID NO: 2; or
ii. a nucleotide sequence which encodes a chitin synthase polypeptide, wherein said polypeptide is at least 98% identical to SEQ ID NO: 4.
12. The method according to claim 6 , wherein said DNA region comprises:
i. a nucleotide sequence which encodes a chitin synthase polypeptide according to SEQ ID NO: 2; or
ii. a nucleotide sequence which encodes a chitin synthase polypeptide, according to SEQ ID NO: 4.
13. The plant cell, plant, fiber, or seed according to any one of claims 1 - 4 or 6 , wherein said promoter is a constitutive promoter.
14. The plant cell, plant, fiber, or seed according to any one of claims 1 - 4 or 6 , wherein said promoter is a fiber-selective promoter.
15. The method according to claim 6 , wherein said promoter is a constitutive promoter.
16. The method according to claim 6 , wherein said promoter is a fiber-selective promoter.