阅读说明：本技术 与玉米增加的能育性相关的遗传基因座 (Genetic loci associated with increased fertility in maize ) 是由 S·W·里奇 S·P·钦塔曼娜尼 M·邓恩 E·S·厄尔索茨 D·J·弗斯特 N·F·马丁 于 2015-02-20 设计创作，主要内容包括：本发明涉及用于鉴别、选择和/或生产具有增加的能育性的玉米植物或植物部分的方法和组合物。还提供了己经通过本发明的方法中的任一种进行鉴别、选择和/或生产的玉米植物或植物部分。(The present invention relates to methods and compositions for identifying, selecting and/or producing maize plants or plant parts with increased fertility. Also provided are maize plants or plant parts that have been identified, selected and/or produced by any of the methods of the invention.)

1. A method for producing a Vip 3A-expressing maize plant or maize germplasm having increased male fertility compared to a Vip 3A-expressing maize plant or maize germplasm having decreased male fertility or being male sterile, the method comprising:

a) providing a first Vip 3A-expressing maize plant or maize germplasm comprising a Quantitative Trait Locus (QTL) located on chromosome 5 that is associated with increased male fertility in a maize plant that expresses Vip3A, wherein the QTL is defined by and includes the base pair (bp) position 80,804,587 to base pair (bp) position 82,325,775 defined by maize B73 Ref Gen _ v2, and further comprises a haplotype comprising a G polymorphism at bp position 80,804,587 and a C polymorphism at bp position 82,325,775;

b) introgressing the QTL of step a) into a second maize plant or maize germplasm; and is

c) Selecting a Vip 3A-expressing maize plant or maize germplasm comprising the QTL, thereby producing a Vip 3A-expressing maize plant or maize germplasm with increased male fertility compared to a Vip 3A-expressing maize plant or maize germplasm without the QTL.

2. The method of claim 1, wherein the QTL comprises a haplotype containing two or more alleles selected from the group consisting of: a C at bp position 80,828,757, a T at bp position 80,829,669, a G at bp position 81,267,278, a T at bp position 81,267,320, a G at bp position 81,763,802, a C at bp position 81,763,824, an a at bp position 81,802,220, a C at bp position 81,802,574, a T at bp position 81,950,558, a T at bp position 81,950,582, a C at bp position 82,085,147, an a at bp position 82,087,667, an a at bp position 82,235,558, a T at bp position 82,236,085, and an a at bp position 82,236,162.

3. The method of claim 1, wherein the QTL comprises SEQ ID NO 259.

4. The method of claim 1, wherein the second maize plant or maize germplasm comprises a vip3A coding sequence.

5. The method of claim 4, wherein the first maize plant or maize germplasm and/or the second maize plant or maize germplasm is hemizygous or homozygous for a vip3A coding sequence.

6. The method of claim 5, wherein the first maize plant or maize germplasm and/or the second maize plant or maize germplasm comprises maize transplantation MIR 162.

7. The method of claim 5, wherein the maize plant produced is hemizygous or homozygous for the vip3A coding sequence.

8. The method of claim 7, wherein the maize plant or maize germplasm produced comprises maize trans plant MIR 162.

9. A method of producing a Vip 3A-expressing maize plant having increased male fertility as compared to a Vip 3A-expressing maize plant having decreased male fertility or being sterile, the method comprising:

a) crossing a first Vip 3A-expressing maize plant with a second maize plant, wherein the first maize plant comprises within its genome a QTL located on chromosome 5 that is associated with increased male fertility in a Vip 3A-expressing maize plant and the second maize plant lacks the QTL, wherein the QTL is defined by and includes a base pair (bp) position 80,804,587 to a base pair (bp) position 82,325,775 as defined by maize B73 Ref Gen _ v2 and further comprises a haplotype comprising a G polymorphism at bp position 80,804,587 and a C polymorphism at bp position 82,325,775; and optionally

b) Backcrossing the resulting Vip 3A-expressing progeny maize plant of step a) comprising the QTL with a parent plant to produce backcrossed progeny plants;

c) selecting progeny plants expressing Vip3A comprising the QTL for backcrossing; and is

d) Performing steps b) and c) at least three times to fix the QTL in the desired genetic background,

thereby producing a Vip 3A-expressing maize plant comprising the QTL and having increased male fertility compared to a Vip 3A-expressing maize plant without the QTL.

10. The method of claim 9, wherein the QTL comprises a haplotype containing two or more alleles selected from the group consisting of: a C at bp position 80,828,757, a T at bp position 80,829,669, a G at bp position 81,267,278, a T at bp position 81,267,320, a G at bp position 81,763,802, a C at bp position 81,763,824, an a at bp position 81,802,220, a C at bp position 81,802,574, a T at bp position 81,950,558, a T at bp position 81,950,582, a C at bp position 82,085,147, an a at bp position 82,087,667, an a at bp position 82,235,558, a T at bp position 82,236,085, and an a at bp position 82,236,162.

11. The method of claim 9, wherein the QTL comprises SEQ ID NO 259.

12. The method of claim 9, wherein the first corn plant and/or the second corn plant is an inbred corn plant or plant part, or an elite corn line.

13. The method of claim 12 wherein the elite maize line is NP2222, NP2660, NP2276, NP2391, NP2460 or ID 3461.

14. The method of claim 9, wherein the QTL is associated with one or more of: increased pollen production, enhanced tassel formation, enhanced anther formation, increased male fertility in plants grown under drought conditions, increased male fertility in plants grown under high temperature conditions at night, or any combination thereof.

15. A maize plant or plant part produced by the method of claim 9.

16. A breeding program comprising the method of claim 9.

17. A method of improving seed production from a maize plant expressing Vip3A, the method comprising:

a) crossing a first maize plant or maize germplasm with a second maize plant or maize germplasm, wherein the first or second maize plant or maize germplasm expresses Vip3A, and wherein the first maize plant or maize germplasm comprises within its genome a QTL on chromosome 5 that is associated with increased male fertility in a maize plant that expresses Vip3A, and the second maize plant or maize germplasm lacks the QTL, wherein the QTL is defined by and includes the base pair (bp) position 80,804,587 to the base pair (bp) position 82,325,775 defined by maize B73 Ref Gen _ v2, and further comprises a haplotype comprising a G polymorphism at bp position 80,804,587 and a C polymorphism at bp position 82,325,775; and is

b) Using a progeny maize plant comprising the QTL as a pollinator in a cross with itself or a second maize plant or maize germplasm that functions as a seed parent,

thereby improving seed production from the cross as compared to a suitable control cross.

18. The method of claim 17, wherein the QTL comprises a haplotype containing two or more alleles selected from the group consisting of: a C at bp position 80,828,757, a T at bp position 80,829,669, a G at bp position 81,267,278, a T at bp position 81,267,320, a G at bp position 81,763,802, a C at bp position 81,763,824, an a at bp position 81,802,220, a C at bp position 81,802,574, a T at bp position 81,950,558, a T at bp position 81,950,582, a C at bp position 82,085,147, an a at bp position 82,087,667, an a at bp position 82,235,558, a T at bp position 82,236,085, and an a at bp position 82,236,162.

19. The method of claim 17, wherein the QTL comprises SEQ ID NO 259.

20. The method of claim 17, wherein the method reduces the ratio of pollen parent to seed parent corn plants required for seed production by at least about 25% compared to a control cross.

21. The method of claim 20, wherein the method increases the number of seeds produced per pollen parent plant and/or seed parent plant by at least about 25% compared to a control cross.

22. The method of claim 20, wherein the progeny is identified by detecting the presence of the QTL in a nucleic acid sample from the progeny or an amplification product thereof.

23. A seed production plan comprising the method of claim 17.

Technical Field

The present invention relates to compositions and methods for identifying, selecting and producing maize plants with increased fertility.

Background

Vip3 protein has been successfully expressed in transgenic plants (e.g., corn and cotton). For example, a hybrid transgenic maize plant can express Vip3A protein at the following levels: this level kills harmful insects and does not negatively affect the plant phenotype. Thus, the Vip3A trait protects the yield and yield potential of hybrid maize plants. However, under normal growth conditions, Vip3 has been observed to cause a reduction in male fertility in certain selfed corn plants. This phenomenon is more pronounced in inbred maize plants that are homozygous for the vip3A transgene. The extent of male fertility reduction is self-specific-some inbred lines exhibit little or no reduction in male fertility when homozygous for the vip3 gene; when homozygous for the Vip3 gene, other inbred lines are somewhat sensitive to Vip3 and exhibit a significant reduction in male fertility; and when homozygous for the Vip3 gene, other inbred lines are highly sensitive to Vip3 and exhibit little or no male fertility. The extent of male fertility reduction is also influenced by environmental factors such as water availability and temperature. In the Vip 3-induced reduction of male fertility, drought and high temperature conditions exacerbate the reduction of male fertility; however, cooler growth conditions have been shown to mitigate the negative effects of Vip3 expression on male fertility.

Identifying a genetic locus that enhances fertility of a maize plant expressing a vip3 transgene may result in more efficient crop production by allowing for the identification, selection, and production of inbred maize plants expressing vip3 with increased male fertility.

Summary of the claimed invention

The present invention provides maize plants with increased male fertility, as well as compositions and methods for identifying, selecting, and producing such plants.

In some embodiments, methods are provided for identifying a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods can include detecting a marker associated with increased male fertility in a maize plant or plant part.

In some embodiments, methods for producing a maize plant having one or more characteristics associated with increased male fertility are provided. Such methods may include: detecting the presence of a marker associated with increased male fertility in a maize plant part, and producing a maize plant from the maize plant part. Such methods may further comprise introducing the marker into the maize plant part.

In some embodiments, breeding methods are provided. Such methods may include: detecting the presence of a marker associated with increased male fertility (e.g., in a nucleic acid (e.g., in an amplification product of a nucleic acid sample from the plant or plant part)) in a maize plant or plant part, and selecting the maize plant or plant part for breeding. In embodiments, the method can further comprise crossing the maize plant (or an ancestor, progeny, or co-generation thereof) with a second maize plant that optionally lacks the marker to produce a progeny maize plant that optionally comprises the marker.

Still further, the present invention provides a method for producing a plant having one or more characteristics associated with increased male fertility, the method comprising: selecting a maize plant from a different population of maize plants comprising a marker associated with increased male fertility as described herein; and crossing the maize plant (or an ancestor, progeny, or equivalent thereof) with itself or a second maize plant to produce a progeny plant comprising the marker, thereby producing a plant having one or more characteristics associated with increased male fertility. In various embodiments, the second maize plant does not comprise the marker. In embodiments, the marker is detected in nucleic acid from the first corn plant and/or progeny plant (e.g., in an amplification product of a nucleic acid sample from the corn plant and/or progeny).

In some embodiments, methods for reducing costs associated with breeding and/or seed production are provided. Such methods may include: detecting the presence of a marker associated with increased male fertility in a maize plant or plant part, and selecting the maize plant or plant part for breeding.

In some embodiments, methods for predicting male fertility are provided. Such methods may include: detecting the presence of a marker associated with increased male fertility (e.g., in a nucleic acid from the plant or plant part) in a maize plant or plant part, wherein the presence of the marker predicts the likelihood of increased male fertility.

In some embodiments, methods are provided for identifying a maize plant or plant part comprising at least one allele associated with increased male fertility. Such methods can include detecting a marker associated with increased male fertility in a maize plant or plant part (e.g., in a nucleic acid from the plant or plant part).

In some embodiments, methods are provided for producing a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods may include: introducing a nucleic acid comprising at least one allele associated with increased male fertility into the genome of a maize plant part, and producing a maize plant from the maize plant part. Such methods may further include: detecting a marker associated with increased male fertility and/or an allele associated with increased male fertility in a nucleic acid (e.g., in a nucleic acid sample) from the maize plant or plant part. In various embodiments, the marker and/or allele is detected in an amplification product from a nucleic acid sample from the maize plant or plant part.

In some embodiments, methods for improving pollen production are provided. Such methods may include: introducing a nucleic acid comprising at least one allele associated with increased pollen production into the genome of a maize plant part, and producing a maize plant from said maize plant part. Such methods may further include: detecting a marker associated with increased pollen production and/or an allele associated with increased pollen production in a nucleic acid (e.g., a nucleic acid sample) from the maize plant part. In many embodiments, the marker is detected in the amplification product of a nucleic acid sample from the plant or plant part.

In some embodiments, methods are provided for producing a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods may include: introducing a genomic region associated with increased male fertility into the genome of a maize plant part, and producing a maize plant from the maize plant part. Such methods may further include: detecting a marker associated with increased male fertility and/or a genomic region associated with increased male fertility in a nucleic acid (e.g., a nucleic acid sample) from the maize plant part. In various embodiments, the marker is detected in the amplification product of a nucleic acid sample from the corn plant or plant part.

In some embodiments, methods for improving pollen production are provided. Such methods may include, consist essentially of, or consist of: introducing a genomic region associated with increased pollen production into the genome of a maize plant part, and producing a maize plant from the maize plant part. Such methods may further include: detecting a marker associated with increased pollen production and/or a genomic region associated with increased pollen production in a nucleic acid (e.g., a nucleic acid sample) from the maize plant part. In various embodiments, the marker is detected in the amplification product of a nucleic acid sample from the corn plant or plant part.

In some embodiments, methods are provided for producing a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods may include: introducing a genomic region comprising one or more transgenes associated with increased male fertility into the genome of a maize plant or plant part.

In some embodiments, methods for improving pollen production are provided. Such methods may include: introducing a genomic region associated with increased pollen production into the genome of a maize plant or plant part, and producing a maize plant from said maize plant part.

In some embodiments, methods are provided for producing a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods may comprise crossing a first corn plant or plant part with a second corn plant or plant part to produce a progeny corn plant or plant part, wherein the first corn plant or plant part comprises within its genome a marker associated with increased male fertility, optionally wherein the second corn plant or plant part lacks the marker, and wherein the progeny corn plant or plant part has the marker within its genome. Such methods may further include: selecting the progeny plant based on the presence of the marker.

In some embodiments, methods are provided for producing a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods can include crossing a first corn plant or plant part with a second corn plant or plant part to produce a progeny corn plant or plant part, wherein the first corn plant or plant part includes within its genome an allele associated with increased male fertility, optionally wherein the second corn plant or plant part lacks the allele, and wherein the progeny corn plant or plant part has the allele within its genome. Such methods may further include: selecting the progeny plant based on the presence of the allele and/or the presence of a marker linked to the allele.

In some embodiments, methods are provided for producing a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods can include crossing a first corn plant or plant part with a second corn plant or plant part to produce a progeny corn plant or plant part, wherein the first corn plant or plant part includes within its genome a genomic region comprising one or more transgenes associated with increased male fertility, optionally wherein the second corn plant or plant part lacks the genomic region, and wherein the progeny corn plant or plant part has the genomic region within its genome. Such methods may further include: selecting the progeny plant based on the presence of the genomic region and/or the presence of a marker linked to the genomic region.

In some embodiments, methods are provided for selecting a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods can include crossing a first corn plant or plant part with a second corn plant or plant part, wherein the first corn plant or plant part includes a marker associated with increased male fertility, and optionally wherein the second corn plant or plant part lacks the marker, and selecting a progeny plant or plant part having the marker.

In some embodiments, methods are provided for selecting a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods can include crossing a first corn plant or plant part with a second corn plant or plant part, wherein the first corn plant or plant part includes an allele associated with increased male fertility, and optionally wherein the second corn plant or plant part lacks the allele, and selecting a progeny plant or plant part having the allele and/or a marker linked to the allele.

In some embodiments, methods are provided for selecting a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods can include crossing a first corn plant or plant part with a second corn plant or plant part, wherein the first corn plant or plant part includes a genomic region comprising one or more transgenes associated with increased male fertility, and optionally wherein said second corn plant or plant part lacks said genomic region, and selecting a progeny plant or plant part having said genomic region and/or a marker linked to said genomic region.

In some embodiments, methods are provided for producing a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods may include: crossing a donor maize plant or plant part with a recurrent maize plant or plant part, and backcrossing progeny to the recurrent maize plant or plant part for one or more generations, wherein the donor maize plant or plant part comprises within its genome a marker associated with increased male fertility, wherein the recurrent maize plant or plant part optionally lacks the marker, and wherein at least one generation in the progeny is identified and/or selected for backcrossing by detecting the presence of the marker.

In some embodiments, methods are provided for producing a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods may include: crossing a donor maize plant or plant part with a recurrent maize plant or plant part, and backcrossing progeny to the recurrent maize plant or plant part for one or more generations, wherein the donor maize plant or plant part comprises within its genome an allele associated with increased male fertility, wherein the recurrent maize plant or plant part optionally lacks the allele, and wherein at least one generation in the progeny is identified and/or selected for backcrossing by detecting the presence of the allele and/or the presence of a marker linked to the allele.

In some embodiments, methods are provided for producing a maize plant or plant part having one or more characteristics associated with increased male fertility. Such methods may include: crossing a donor maize plant or plant part with a recurrent maize plant or plant part, and backcrossing progeny with the recurrent maize plant or plant part for one or more generations, wherein the donor maize plant or plant part comprises within its genome a genomic region comprising one or more transgenes associated with increased male fertility, wherein the recurrent maize plant or plant part optionally lacks the genomic region, and wherein at least one generation in the progeny is identified and/or selected for backcrossing by detecting the presence of the genomic region and/or the presence of a marker linked to the genomic region.

In various embodiments, the present invention also provides methods for improving seed production from a corn plant. To illustrate, in a representative embodiment, the present invention provides a method for improving seed production from a corn plant, the method comprising: crossing a first maize plant with a second maize plant, wherein the first maize plant comprises within its genome a marker associated with increased male fertility as described herein, and the second maize plant optionally lacks the marker, to produce a progeny maize plant comprising the marker; and using a progeny maize plant comprising the marker as a pollinator in a cross with itself or a second maize plant that functions as a seed parent (e.g., crossing the progeny plant comprising the marker with itself, wherein the progeny plant functions as a pollinator and as a seed parent, or crossing the progeny plant comprising the progeny plant with a second maize plant, wherein the progeny plant functions as a pollinator and the second maize plant functions as a seed parent), thereby increasing seed production from the cross as compared to a suitable control cross.

In representative embodiments of the methods of the invention, the marker, allele, haplotype and/or genomic region comprises, consists essentially of or consists of:

(a) one or more markers located within one or more of the chromosomal intervals described in table 1, or a marker/allele/haplotype/genomic region positioned 10cM or less therefrom;

(b) one or more of the desired alleles described in table 2, table 8 and/or table 10, or a marker/allele/haplotype/genomic region positioned 10cM or less therefrom;

(c) a haplotype comprising two or more of the desired alleles described in table 2, table 8 and/or table 10, or a marker/allele/haplotype/genomic region positioned 10cM or less therefrom;

(d) an allele or haplotype that is in linkage disequilibrium with one or more of the chromosomal intervals described in table 1, or a marker/allele/haplotype/genomic region positioned 10cM or less therefrom;

(e) an allele or haplotype that is in linkage disequilibrium with one or more of the desired alleles described in table 2, table 8 and/or table 10, or a marker/allele/haplotype/genomic region positioned 10cM or less therefrom;

(f) an allele or haplotype that is in linkage disequilibrium with a haplotype comprising two or more of the desired alleles described in table 2, table 8 and/or table 10, or a marker/allele/haplotype/genomic region positioned 10cM or less therefrom; or

(g) Any combination of (a) to (f).

In further embodiments of the methods of the invention, the marker/allele/haplotype/genomic region is located within:

(a) chromosomal interval 1 as described in table 1;

(b) chromosomal interval 2 as described in table 1;

(c) one or more of chromosomal intervals 3-17 as described in table 1;

(d) one or more of chromosomal intervals 18-45 as described in table 1;

(e) one or more of chromosome intervals 46 through 12743 as described in table 1;

(f) one or more of chromosome intervals 12744 to 12749 as described in table 1;

(g) one or more of chromosome intervals 12750 to 12755 as described in table 1;

(h) chromosome interval 12756 as described in table 1;

(i) one or more of chromosome intervals 12757-12762 as described in table 1;

(j) one or more of chromosome intervals 12763 through 12768 as described in table 1;

(k) one or more of chromosomal intervals 6 to 9371 as described in table 1;

(l) Chromosomal interval 3 as described in table 1;

(m) a chromosomal interval 13 as described in table 1;

(n) a chromosomal interval 25 as described in table 1; or

(o) any combination of (a) to (n).

In some embodiments, the marker/allele/haplotype/genomic region comprises, consists essentially of, or consists of one or more of the following desired alleles:

(a) on chromosome 5 as described in table 8;

(b) on chromosome 3 as described in table 8;

(c) on chromosome 7 as described in table 8;

(d) on chromosome 10 as described in table 8;

(e) on chromosomes 3 and 5 as described in table 8;

(f) on chromosomes 5 and 7 as described in table 8;

(g) on chromosomes 3, 5 and 7 as described in table 8;

(h) on chromosomes 5 and 10 as described in table 8;

(i) on chromosomes 3, 5 and 10 as described in table 8;

(j) any combination of (a) to (i); or

(k) Or a marker/allele/haplotype/genomic region positioned 10cM or less from any one of (a) to (j).

In other representative embodiments, the marker/allele/haplotype/genomic region comprises a haplotype comprising two or more of the following desired alleles:

(a) on chromosome 5 (QTL 5.1 and/or QTL 5.2) as described in table 8;

(b) on chromosome 3 as described in table 8;

(c) on chromosome 7 as described in table 8;

(d) on chromosome 10 as described in table 8;

(e) on chromosomes 3 and 5 as described in table 8;

(f) on chromosomes 5 and 7 as described in table 8;

(g) on chromosomes 3, 5 and 7 as described in table 8;

(h) on chromosomes 5 and 10 as described in table 8;

(i) on chromosomes 3, 5 and 10 as described in table 8;

(j) any combination of (a) to (i); or

(k) Or a marker/allele/haplotype/genomic region positioned 10cM or less from any one of (a) to (j).

In some embodiments, non-naturally occurring maize plants and plant parts (e.g., maize plants and plants identified, selected, and/or produced according to the methods of the invention) that include one or more markers, alleles, and/or genomic regions associated with increased male fertility are provided.

In some embodiments, progeny and plant parts derived from maize plants and plant parts (e.g., maize plants and plants identified, selected, and/or produced according to the methods of the invention) that include one or more markers, alleles, and/or genomic regions associated with increased male fertility are provided.

In some embodiments, isolated and/or purified markers associated with increased male fertility are provided. Such indicia may include, consist essentially of, or consist of: 1-350, a reverse complement thereof, an informative or functional fragment thereof.

In some embodiments, isolated and/or purified Quantitative Trait Loci (QTLs) associated with increased male fertility are provided. Such QTLs may comprise, consist essentially of, or consist of one or more of the chromosome fragments described in table 1.

In some embodiments, amplification products comprising one or more markers, alleles and/or genomic regions associated with increased male fertility are provided. Such amplification products may comprise, consist essentially of, or consist of: 1 to 350, the reverse complement thereof, or an informative or functional fragment thereof.

In some embodiments, probes for detecting one or more markers, alleles and/or genomic regions associated with increased male fertility are provided (e.g., as described in table 9). Such probes may comprise, consist essentially of, or consist of: 526 to 613, the reverse complement thereof, or an informative or functional fragment thereof.

The foregoing and other objects and aspects of the present invention are explained in detail in the drawings and description set forth below.

Brief description of the tables

Table 1 describes the segments of maize chromosomes 1, 3,4, 5,6, 7,8, 9, and 10.

Table 2 describes the alleles of interest located on chromosome 5 in maize.

Table 3 describes the proteins of interest encoded by maize chromosome 5, along with fragments of chromosome 5 that encode those proteins.

Table 4 describes Vip3 proteins and nucleic acid sequences encoding those proteins.

Table 5 shows that the decrease in male fertility induced by Vip3 varies across genetic backgrounds.

Table 6 shows LOD (LOD) scores for identifying QTLs (quantitative trait loci) using F2 plants from a parental cross of NP2222 (described in U.S. patent No. 6,710,233) with NP2276 (described in U.S. patent No. 6,706,955) and a parental cross of ID3461 (described in international patent application No. WO 2009142752) with NP 2276.

Table 7 shows the fertility index used to score F2 plants from the crossover of NP2222 with NP2276 and the crossover of ID3461 with NP 2276.

Table 8 describes QTLs associated with increased anther number and/or improved anther quality in maize plants expressing Vip3 protein.

Table 9 describes exemplary nucleic acid probes and primers that can be used to identify favorable alleles of the QTLs described in table 8.

Table 10 describes SNPs present in the inbred maize line NP2222 (comparative maize line NP2276) across the male fertility-associated QTL interval in table 8. Where the alleles differ between the two strains, the NP2222 allele corresponds to the desired (favorable) allele.

Table 11 provides public lines predicted to contain favorable alleles.