Compositions and methods for rapid and efficient production of quality protein maize

Patent metadata

Field	Value
Publication number	US-9603317-B2
Application number	US-201213362641-A
Country	US
Kind code	B2
Filing date	Jan 31, 2012
Priority date	Jan 31, 2011
Publication date	Mar 28, 2017
Grant date	Mar 28, 2017

Abstract

Compositions and methods for the universal and accelerated production of QPM are disclosed.

First claim

What is claimed is:

1. A method for rapid and accelerated breeding of quality protein maize (QPM) comprising:
a) providing seeds from a plant comprising an RNAi construct which acts in a dominant fashion to down modulate expression of 19 and 22 kDa alpha zeins, said down modulation decreasing the vitreous nature of seeds obtained from said plant, said RNAi construct comprising a nucleic acid encoding a selectable marker and a visible reporter gene, said visible reporter gene facilitating identification of cells comprising said RNAi construct;
b) propagating said seed from a) and crossing the resulting plant with a QPM plant line;
c) obtaining seeds from plants resulting from said cross and harvesting those kernels which exhibit an increased kernel vitreousness and the signal produced by the visible reporter gene when compared to seeds in a) thereby providing a dominant selectable phenotype for elevated lysine and rapid selection of QPM, the method comprising crossing plants obtained from the kernels of c) with a non-QPM line thereby promoting introgression of the QPM trait into said non-QPM line further comprising breeding a transgenic plant from the harvested kernels to yield a progeny plant that has an increase in the amount of at least one amino acid as a dominant trait, said method comprising backcrossing said progeny plant to said non-QPM parent containing a traditional o2 mutation to eliminate the RNAi transgene construct.

2. A method for production of improved maize seed comprising:
a) providing seeds from a plant comprising an RNAi construct which acts in a dominant fashion to down modulate expression of 19 and 22 kDa alpha zeins, said down modulation decreasing the vitreous nature of seeds obtained from said plant, said RNAi construct comprising a nucleic acid encoding a selectable marker and a visible reporter gene; wherein said visible reporter gene is used for identification of transformed seeds;
b) propagating said seeds which exhibit expression of said visible reporter marker gene from a) and crossing the resulting plant with an Illinois High Protein (IHP) line;
c) obtaining seeds from plants resulting from said cross and harvesting those kernels which exhibit expression of said visible reporter gene, said seeds exhibiting an increased kernel vitreousness, and elevated protein content when compared to seeds in a) or seeds obtained from an Illinois High Protein line lacking said RNAi construct, wherein said seed have the genotype IHP/−; P6z1RNAi/−.

3. The method of claim 2 , wherein said seed from step c) exhibits elevated lysine content.

4. The method of claim 2 , wherein said seed from step c) exhibits elevated tryptophan content.

5. The method of claim 2 , wherein said seed from step c) exhibits elevated methionine content.

6. The method of claim 2 , wherein said visible reporter gene is green fluorescent protein.

7. The method of claim 2 , wherein said RNAi construct is shown in FIG. 6 .

8. The method of claim 1 , wherein said visible reporter gene is green fluorescent protein (GFP) and said selectable marker gene is bar.

9. The method of claim 2 , wherein said visible reporter gene is green fluorescent protein (GFP) and said selectable marker is bar.

Assignees

• Messing Joachim
• Wu Yongrul
• Univ Rutgers

Inventors

• Messing Joachim
• Wu Yongrul

Classifications

• C07K14/425

  Zeins · CPC title

• C12N15/8218

  Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS] · CPC title

• C12N15/8251

  Amino acid content, e.g. synthetic storage proteins, altering amino acid biosynthesis · CPC title

• A01H5/10

  Seeds · CPC title

• A01H1/02 Primary

  Methods or apparatus for hybridisation; Artificial pollination {; Fertility} · CPC title