IP Library Granted Patent US 8,252,975
Granted Patent B2
US 8,252,975 · App. 12/524,542 · Granted Aug 28, 2012

Genetically modified plants which synthesize a low amylose starch with increased swelling power

View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 8,252,975
App. No.
12/524,542
Granted
Aug 28, 2012
Kind
B2
Abstract

The present invention relates to genetically modified monocotyledonous plant cells and plants whose starch has an apparent amylose content of less than 5% by weight and an increased activity of a protein with the activity of a starch synthase II and an increased activity of a protein with the activity of a glucan, water dikinase. Such plants synthesize starch with an increased hot-water swelling power. Methods and processes for the generation/preparation of these plant cells, plants, starches and flours are likewise subject matter of the present invention.

Claims (56)

1. A genetically modified monocotyledonous plant cell that comprises a waxy mutation that results in the synthesis of a starch with an apparent amylose content of less than 5% by weight and that comprises a foreign nucleic acid molecule that increases the expression of a starch synthase II, and a foreign nucleic acid molecule that increases the expression of a glucan water dikinase, each compared to a corresponding monocotyledonous plant cell that does not comprise said foreign nucleic acid molecules, wherein said starch has a hot-swelling power of between 60 to 100 g/g.

2. A monocotyledonous plant comprising the genetically modified plant cell of claim 1 .

3. The monocotyledonous plant of claim 2 , wherein said plant is rice, maize or wheat.

4. A propagation material of monocotyledonous plants, wherein said propagation material comprises the genetically modified plant cell of claim 1 .

5. A method of generating a genetically modified monocotyledonous plant, said method comprising the following steps:

a) genetically modifying a monocotyledonous plant cell comprising the following steps:

i) introducing, into the plant cell, a foreign nucleic acid molecule that increases the expression of a starch synthase II in comparison with a corresponding wild-type plant cell that does not comprise said foreign nucleic acid molecule,

ii) introducing, into the plant cell, a foreign nucleic acid molecule that increases the expression of a glucan water dikinase in comparison with a corresponding wild-type plant cell that does not comprise said foreign nucleic acid molecule,

iii) introducing, into the plant cell, a foreign nucleic acid molecule that reduces the expression of a GBSSI in comparison with a corresponding wild-type plant cells that does not comprise said foreign nucleic acid molecule,

where steps i to iii can be carried out in any sequence, individually or simultaneously; and

b) regenerating a plant from the plant cell of step a).

6. A process for the preparation of a modified starch, said process comprising the step of extracting the starch from the genetically modified plant cell of claim 1 .

7. A process for the preparation of flours, said process comprising the step of grinding parts of the plant of claim 2 .

8. The method of claim 5 , further comprising the following steps:

c) generating further plants by isolating plant cells from a plant of step b) and repeating steps a) and b); and

d) repeating step c) until a plant has been generated which has an increased expression of a starch synthase II, an increased expression of a glucan water dikinase, and a reduced expression of a GBSSI, in comparison with a corresponding wild-type plant that has not been genetically modified by introducing the foreign nucleic acid molecules of claim 5 .

9. A process for the preparation of flours, said process comprising the step of grinding parts of the propagation material of claim 4 .

10. A process for the preparation of flours, said process comprising the step of grinding parts of the plants produced by the method of claim 5 .

11. The genetically modified monocotyledonous plant cell of claim 1 , wherein the plant cell comprises

at least one foreign nucleic acid molecule encoding a starch synthase II; and

at least one foreign nucleic acid molecule encoding a glucan water dikinase.

12. The genetically modified monocotyledonous plant cell of claim 11 , wherein the at least one foreign nucleic acid molecule encoding a starch synthase II comprises:

i) a nucleic acid molecule encoding a protein comprising the amino acid sequence of SEQ. ID. NOs.: 4 or 6;

ii) a nucleic acid molecule comprising the nucleic acid sequence of SEQ. ID. NOs.: 3 or 5; or

iii) a nucleic acid molecule that hybridizes under stringent conditions with at least one strand of the nucleic acid molecule described in ii), wherein the stringent conditions are:

hybridization buffer: 2×SSC, 10×Denhardt solution (Ficoll 400+PEG+BSA; ratio 1:1:1), 0.1% SDS; 5 mM EDTA, 50 mM Na 2 HPO4, 250 μg/ml of herring sperm DNA, 50 μg/ml of tRNA, or 25 M sodium phosphate buffer pH 7.2; 1 mM EDTA; 7% SDS;

hybridization temperature: T=65 to 68° C.;

wash buffer: 0.1×SSC; 0.1% SDS; and

wash temperature: T=65 to 68° C.

13. The genetically modified monocotyledonous plant cell of claim 11 , wherein the at least one foreign nucleic acid molecule encoding a glucan water dikinase comprises:

i) a nucleic acid molecule encoding a protein comprising the amino acid sequence of SEQ. ID. NO.: 2;

ii) a nucleic acid molecule comprising the nucleic acid sequence of SEQ. ID. NO.: 1; or

iii) a nucleic acid molecule that hybridizes under stringent conditions with at least one strand of the nucleic acid molecule described in ii), wherein the stringent conditions are:

hybridization buffer: 2×SSC, 10×Denhardt solution (Ficoll 400+PEG+BSA; ratio 1:1:1), 0.1% SDS; 5 mM EDTA, 50 mM Na 2 HPO4, 250 μg/ml of herring sperm DNA, 50 μg/ml of tRNA, or 25 M sodium phosphate buffer pH 7.2; 1 mM EDTA; 7% SDS;

hybridization temperature: T=65 to 68° C.;

wash buffer: 0.1×SSC; 0.1% SDS; and

wash temperature: T=65 to 68° C.

14. A genetically modified monocotyledonous plant cell comprising a foreign nucleic acid molecule that reduces the expression of an endogenous GBSSI, a foreign nucleic acid molecule that increases expression of a starch synthase II, and a foreign nucleic acid molecule that increases expression of a glucan water dikinase, each compared to a corresponding monocotyledonous plant cell that does not comprise said foreign nucleic acid molecules, wherein said plant cell synthesizes a starch having a hot-swelling power of between 60 to 100 g/g.

15. The genetically modified monocotyledonous plant cell of claim 14 , wherein the foreign nucleic acid molecule that reduces the expression of an endogenous GBSSI comprises:

i) a nucleic acid molecule encoding a protein comprising the amino acid sequence of SEQ. ID. NO.: 8, 10, or 12;

ii) a nucleic acid molecule comprising the nucleic acid sequence of SEQ. ID. NO.: 7, 9, or 11; or

iii) a nucleic acid molecule that hybridizes under stringent conditions with at least one strand of the nucleic acid molecule described in ii), wherein the stringent conditions are:

hybridization buffer: 2×SSC, 10×Denhardt solution (Ficoll 400+PEG+BSA; ratio 1:1:1), 0.1% SDS; 5 mM EDTA, 50 mM Na 2 HPO4, 250 μg/ml of herring sperm DNA, 50 μg/ml of tRNA, or 25 M sodium phosphate buffer pH 7.2; 1 mM EDTA; 7% SDS;

hybridization temperature: T=65 to 68° C.;

wash buffer: 0.1×SSC; 0.1% SDS; and

wash temperature: T=65 to 68° C.

16. The genetically modified monocotyledonous plant cell of claim 14 , wherein the foreign nucleic acid molecule that reduces the expression of an endogenous GBSSI comprises

(i) a nucleic acid molecule encoding at least one antisense RNA that reduces the expression of at least one endogenous gene encoding a GBSSI protein;

(ii) a nucleic acid molecule which, via a co-suppression effect, reduces the expression of at least one endogenous gene encoding a GBSSI protein; or

(iii) a nucleic acid molecule that simultaneously encodes at least one antisense RNA and at least one sense RNA, where said antisense RNA and said sense RNA form a double-stranded RNA molecule that reduces the expression of at least one endogenous gene encoding a GBSSI protein.

17. A method of generating a genetically modified monocotyledonous plant comprising the following steps:

a) genetically modifying a monocotyledonous plant cell that comprises a waxy mutation that results in the synthesis of a starch having an apparent amylose content of less than 5% comprising the following steps:

i) introducing, into the plant cell, a foreign nucleic acid molecule that increases the expression of a starch synthase II in comparison with a corresponding wild-type plant cell that does not comprise said foreign nucleic acid molecule, and

ii) introducing, into the plant cell, a foreign nucleic acid molecule that increases the expression of a glucan, water dikinase in comparison with a corresponding wild-type plant cell that does not comprise said foreign nucleic acid molecule

where steps i to ii can be carried out in any sequence, individually or simultaneously; and

b) regenerating a plant from the plant cell of step a).

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 9, 2019
From: BAYER CROPSCIENCE AKTIENGESELLSCHAFT; BAYER BIOSCIENCE GMBH; BAYER INTELLECTUAL PROPERTY GMBH
To: BASF AGRICULTURAL SOLUTIONS SEED, US LLC
Reel/Frame 049138/0323 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 2, 2015
From: BAYER CROPSCIENCE AG
To: BAYER INTELLECTUAL PROPERTY GMBH
Reel/Frame 035355/0836 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 28, 2009
From: FROHBERG, CLAUS; SCHMIDT, RALF-CHRISTIAN
To: BAYER CROPSCIENCE AG
Reel/Frame 023016/0008 →