MAIZE CYTOPLASMIC MALE STERILITY (CMS) S-TYPE RESTORER Rf3 GENE, MOLECULAR MARKERS AND THEIR USE
The present disclosure provides a method for selecting a plant comprising a functional restorer gene for maize S-type cytoplasmic male sterility comprising the steps of (a) screening a population of plants for at least one marker nucleic acid, wherein the marker nucleic acid comprises an allele linked to the functional restorer gene for maize S-type cytoplasmic male sterility; (b) detecting the marker nucleic acid; (c) identifying a plant comprising the marker nucleic acid; and (d) selecting the plant comprising the marker nucleic acid, wherein the plant comprising the marker nucleic acid further comprises the functional restorer gene for maize S-type cytoplasmic male sterility. The present disclosure also provides methods for restoring fertility in a progeny of an S-type cytoplasmic male sterile plant and methods for transferring an Rf3 gene into a progeny plant.
1 .- 32 . (canceled)
33 . A method for transferring an Rf3 gene into a progeny maize plant, the method comprising the steps of:
(a) crossing a first parent maize plant and a second parent maize plant to produce a progeny maize plant, wherein at least one parent maize plant comprises the Rf3 gene;
(b) analyzing the progeny maize plant for the presence of the Mo17-14388 marker or the DASCMS-SRf39 marker that is linked to the Rf3 gene to obtain an Rf3 progeny maize plant;
(c) backcrossing the Rf3 progeny maize plant with either the first parent maize plant or the second parent maize plant to produce a next-generation progeny maize plant; and
(d) analyzing the next-generation progeny maize plant for the presence of the Mo17-14388 marker or the DASCMS-SRf39 marker that is linked to the Rf3 gene to obtain an Rf3 next-generation progeny maize plant.
34 .- 35 . (canceled)
36 . The method of claim 33 , wherein the marker nucleic acid Mo17-14388 is selected from the group consisting of SEQ ID NO:1, SEQ ID NO: 12, and SEQ ID NO: 23, and any combination thereof.
37 . (canceled)
38 . The method of claim 33 , wherein the marker nucleic acid DASCMS-SRf39 is selected from the group consisting of SEQ ID NO: 86, SEQ ID NO: 87, and SEQ ID NO: 88, and any combination thereof.
39 .- 40 . (canceled)
41 . The method of claim 33 , wherein the Mo17-14388 is located on chromosome 2 within 8.3 Mb of the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility.
42 . The method of claim 33 , wherein the Mo17-14388 is located on chromosome 2 within 0.5 Mb of the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility.
43 . The method of claim 33 , wherein the Mo17-14388 is located on chromosome 2 within 100 kb of the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility.
44 . The method of claim 33 , wherein the maize plant is a hybrid maize plant.
45 . The method of claim 33 , wherein the progeny maize plant belongs to the Stiff Stalk heterotic group.
46 . The method of claim 33 , wherein the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility consists essentially of SEQ ID NO:92.
47 . The method of claim 33 , wherein the marker nucleic acid Mo17-14388 is located on chromosome 2 within 1.3 Mb of the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility.
48 . The method of claim 33 , wherein the DASCMS-SRf39 is located on chromosome 2 within 8.3 Mb of the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility.
49 . The method of claim 33 , wherein the marker nucleic acid DASCMS-SRf39 is located on chromosome 2 within 1.3 Mb of the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility.
50 . The method of claim 33 , wherein the DASCMS-SRf39 is located on chromosome 2 within 0.5 Mb of the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility.
51 . The method of claim 33 , wherein the DASCMS-SRf39 is located on chromosome 2 within 100 kb of the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility.
52 . The method of claim 33 , the method further comprising introducing the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility into the second parent maize plant.
53 . The method of claim 52 , wherein the introducing is selected from the group consisting of transformation, crossing, backcrossing, homologous recombination, and mutagenesis.
54 . The method of claim 33 , said screening comprising the steps of:
isolating nucleic acid molecules from the population of progeny maize plants;
contacting the isolated nucleic acid molecules with a set of oligonucleotides; and
amplifying the isolated nucleic acid molecules and the oligonucleotides to produce an amplicon, wherein the amplicon comprises a detectable signal that is indicative of the presence of the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility.
55 . The method of claim 46 , wherein the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility is comprised in an organism selected from the group consisting of a plant, a yeast, a bacterium, and a virus.
56 . The method of claim 55 , wherein the functional Rf3 restorer gene for maize S-type cytoplasmic male sterility is comprised in a plant cell.
57 . The method of claim 44 , wherein the first parent maize plant or the second parent maize plant of the hybrid maize plant belongs to the Stiff Stalk heterotic group.