Cotton fibers with increased glucosamine content
An isolated nucleic acid molecule is provided comprising a nucleotide sequence which encodes a glutamine:fructose-6-phosphate amidotransferase from E. coli which is particularly suitable for expression in cotton plant cells. The invention also relates to plant cells or plants, in particular to cotton plant cells or cotton plants which produce an increased amount of positively charged polysaccharides in their cell walls. Furthermore methods and means are provided to increase the content of positively charged polysaccharides in the cell walls of cotton cells, in particular in cotton fibers. Fibers obtained from such cotton plants have an altered chemical reactivity which can be used to attach reactive dyes or other textile finish reagents to these fibers.
1. A cotton plant cell comprising a chimeric gene comprising the following operably linked DNA regions:
a) a plant-expressible promotor,
b) a DNA region coding for a GFAT polypeptide wherein said GFAT is encoded by a nucleotide sequence comprising
i) a nucleotide sequence according to SEQ ID NO: 1,
ii) or a variant thereof, wherein one or more nucleotides differ from the nucleotide sequence according to SEQ ID NO: 1, provided that said variant differs in no more than 20 nucleotides from SEQ ID NO: 1, which encodes a glutamine:fructose-6-phosphate-amidotransferase (GFAT) according to SEQ ID NO: 2
iii) or a complementary sequence of i) or ii)), and
c) optionally a DNA region involved in transcription termination and polyadenylation
wherein said cotton plant cell additionally comprises a second chimeric gene comprising the following operably linked DNA regions:
a) a plant-expressible promotor,
b) a DNA sequence coding for a chitin synthase polypeptide and
c) optionally a DNA region involved in transcription termination and polyadenylation.
2. A cotton plant cell according to claim 1 wherein said chitin synthase is an N-acetylglucosamine transferase of the Nod C type.
3. A cotton plant cell according to claim 2 wherein said chitin synthase polypeptide comprises a Golgi localization signal.
4. A cotton plant consisting of plant cells according to claim 1 .
5. A method for producing cotton fibers with positively charged polysaccharides, such as oligo-N-acetylglucosamines or oligo-glucosamines, comprising the steps of
i) expressing a chimeric gene comprising the following operably linked DNA regions:
a) a plant-expressible promotor,
b) a DNA region coding for a GFAT polypeptide wherein said GFAT is encoded by a nucleotide sequence comprising
i) a nucleotide sequence according to SEQ ID NO: 1,
ii) or a variant thereof, wherein one or more nucleotides differ from the nucleotide sequence according to SEQ ID NO: 1, provided that said variant litters in no more than 20 nucleotides from SEQ ID NO: 1, which encodes a glutamine:fructose-6-phosphate-amidotransferase (GFAT) according to SEQ ID NO: 2
iii) or a complementary sequence of i) or ii)), and
c) optionally a DNA region involved in transcription termination and polyadenylation
and expressing a second chimeric gene comprising
a) a plant-expressible promotor,
b) a DNA sequence coding for a chitin synthase polypeptide and
c) optionally a DNA region involved in transcription termination and polyadenylation
in a cotton plant cell,
ii) regenerating a cotton plant from cotton plant cells of step i) and
iii) optionally isolating fibers from said cotton plant.
6. The cotton plant cell according to claim 1 , wherein said plant-expressible promotor is a fiber-preferential promotor.
7. The method according to claim 5 , wherein said plant-expressible promotor is a fiber-preferential promotor.