Generating maize plants with enhanced resistance to northern leaf blight
Compositions and methods for generating maize plants that exhibit resistance to northern leaf blight are provided herein. Isolated polynucleotides encoding a polypeptide that confers resistance to northern leaf blight, polynucleotide constructs comprising such, and maize plants comprising the polynucleotide constructs are provided. The methods include expressing an isolated polynucleotide in a maize cell via standard transformation methods and obtaining a maize plant from said maize cell.
1. A maize plant cell comprising a heterologous polynucleotide construct comprising an isolated polynucleotide operably linked to a regulatory element, wherein the isolated polynucleotide comprises:
a. the nucleotide sequence set forth in SEQ ID NO:1 (PH4GP c-DNA), SEQ ID NO:3 (PH1W2 cDNA), or SEQ ID NO:9 (PH4GP Genomic sequence);
b. a nucleotide sequence encoding a CC-NB-LRR polypeptide comprising an amino acid sequence of at least 95% sequence identity to SEQ ID NO:2 or SEQ ID NO:4, wherein the CC-NB-LRR polypeptide comprises SEQ ID NO:10 at the positions corresponding to amino acids 633-648 of SEQ ID NO:2 or SEQID NO:4, respectively; or
c. a nucleotide sequence encoding a CC-NB-LRR polypeptide comprising the sequence set forth in SEQ ID NO:2 or SEQ ID NO:4.
2. The maize plant cell of claim 1 , wherein the polynucleotide construct further comprises one or more additional heterologous nucleic acid sequences that encode a polypeptide selected from the group consisting of: a polypeptide conferring disease resistance, a polypeptide conferring herbicide resistance, a polypeptide conferring insect resistance, a polypeptide involved in carbohydrate metabolism, a polypeptide involved in fatty acid metabolism, a polypeptide involved in amino acid metabolism, a polypeptide involved in plant development, a polypeptide involved in plant growth regulation, a polypeptide involved in yield improvement, a polypeptide involved in drought resistance, a polypeptide involved in cold resistance, a polypeptide involved in heat resistance, and/or a polypeptide involved in salt resistance, wherein each heterologous nucleic acid sequence is operably linked to a promoter.
3. The maize plant cell of claim 2 , wherein a polypeptide conferring disease resistance is a polypeptide that confers resistance to northern leaf blight (NLB).
4. The maize plant cell of claim 3 , wherein said polypeptide conferring resistance to northern leaf blight is a polypeptide having an amino acid sequence of at least 90% sequence identity when compared to SEQ ID NO:11 or 12.
5. A maize plant comprising the maize plant cell of claim 1 .
6. A method for producing a maize plant that exhibits increased resistance to northern leaf blight (NLB) comprising,
expressing in the plant a heterologous polynucleotide construct comprising a polynucleotide operably linked to at least one regulatory sequence, wherein the polynucleotide comprises:
i. a nucleotide sequence set forth in SEQ ID NO:1 (PH4GP c-DNA), SEQ ID NO:3 (PH1W2 cDNA), or SEQ ID NO:9 (PH4GP Genomic sequence);
ii. a nucleotide sequence encoding a CC-NB-LRR polypeptide comprising an amino acid sequence of at least 95% sequence identity when compared to SEQ ID NO:2 or SEQ ID NO:4, wherein said polypeptide comprises the amino acid sequence set forth in SEQ ID NO:10;
iii. a nucleotide sequence encoding a CC-NB-LRR polypeptide having the sequence set forth in SEQ ID NO:2 or SEQ ID NO:4; or
iv. the nucleotide sequence complementary to (i), (ii), or (iii); and thereby generating a maize plant that exhibits increased resistance to northern leaf blight, wherein said maize plant comprises in its genome the heterologous polynucleotide construct.
7. The method of claim 6 , wherein said at least one regulatory sequence is a promoter.
8. The method of claim 6 , wherein said at least one regulatory sequence is a terminator.
9. The method of claim 6 , wherein said regulatory sequence is native to maize.
10. The method of claim 6 , wherein said polynucleotide construct comprises one or more additional heterologous nucleic acid sequences that encode a polypeptide selected from the group consisting of: a polypeptide conferring disease resistance, a polypeptide conferring herbicide resistance, a polypeptide conferring insect resistance, a polypeptide involved in carbohydrate metabolism, a polypeptide involved in fatty acid metabolism, a polypeptide involved in amino acid metabolism, a polypeptide involved in plant development, a polypeptide involved in plant growth regulation, a polypeptide involved in yield improvement, a polypeptide involved in drought resistance, a polypeptide involved in cold resistance, a polypeptide involved in heat resistance, and/or a polypeptide involved in salt resistance, wherein each heterologous nucleic acid sequence is operably linked to a promoter.
11. The method of claim 10 , wherein the polypeptide conferring disease resistance is a polypeptide that confers resistance to northern leaf blight (NLB).
12. The method of claim 11 , wherein said polypeptide conferring resistance to northern leaf blight is a polypeptide having an amino acid sequence of at least 90% sequence identity when compared to SEQ ID NO:11 or 12.
13. A method of obtaining a maize plant that exhibits resistance to northern leaf blight (NLB), said method comprising,
a. crossing a maize plant generated by the method of claim 6 with a maize plant that does not comprise in its genome the polynucleotide construct;
b. obtaining a progeny plant that exhibits resistance to northern leaf blight, wherein said progeny plant comprises the polynucleotide construct in its genome.
14. The maize plant cell of claim 1 , wherein the isolated polynucleotide comprises a nucleotide sequence encoding a CC-NB-LRR polypeptide comprising an amino acid sequence of at least 95% sequence identity to SEQ ID NO:2 or SEQ ID NO:4, wherein the CC-NB-LRR polypeptide comprises SEQ ID NO:10 at the positions corresponding to amino acids 633-648 of SEQ ID NO:2 or SEQID NO:4, respectively.
15. The maize plant cell of claim 1 , wherein the isolated polynucleotide comprises a nucleotide sequence encoding a CC-NB-LRR polypeptide comprising the sequence set forth in SEQ ID NO:2 or SEQ ID NO:4.
16. The method of claim 6 , wherein the polynucleotide operably linked to at least one regulatory sequence comprises a nucleotide sequence encoding a CC-NB-LRR polypeptide comprising an amino acid sequence of at least 95% sequence identity when compared to SEQ ID NO:2 or SEQ ID NO:4, wherein said polypeptide comprises the amino acid sequence set forth in SEQ ID NO:10.
17. The method of claim 6 , wherein the polynucleotide operably linked to at least one regulatory sequence comprises a nucleotide sequence encoding a CC-NB-LRR polypeptide having the sequence set forth in SEQ ID NO:2 or SEQ ID NO:4.
18. The maize plant cell of claim 1 , wherein the isolated polynucleotide comprises the nucleotide sequence set forth in SEQ ID NO:9.
19. The method of claim 6 , wherein the polynucleotide operably linked to at least one regulatory sequence comprises the nucleotide sequence set forth in SEQ ID NO:9.