The stiff-stalk heterotic group in Maize (Zea mays L.) is an important source of inbreds used in U.S. commercial hybrid production. Founder inbreds B14, B37, B73, and, to a lesser extent, B84, are found in the pedigrees of a majority of commercial seed parent inbred lines. We created high-quality genome assemblies of B84 and four expired Plant Variety Protection (ex-PVP) lines LH145 representing B14, NKH8431 of mixed descent, PHB47 representing B37, and PHJ40, which is a Pioneer Hi-Bred International (PHI) early stiff-stalk type. Sequence was generated using long-read sequencing achieving highly contiguous assemblies of 2.13-2.18 Gbp with N50 scaffold lengths >200 Mbp. Inbred-specific gene annotations were generated using a

The Stiff Stalk heterotic pool is a foundation of US maize seed parent germplasm and has been heavily utilized by both public and private maize breeders since its inception in the 1930’s. Flowering time and plant height are critical characteristics for both inbred parents and their test crossed hybrid progeny. To study these traits, a six parent multiparent advanced generation intercross (MAGIC) population was developed including maize inbred lines B73, B84, PHB47 (B37 type), LH145 (B14 type), PHJ40 (novel early Stiff Stalk), and NKH8431 (B73/B14 type). A set of 779 doubled haploid lines were evaluated for flowering time and plant height in two field replicates in 2016 and 2017, and a subset of 689 and 561 doubled haploid lines were crossed to two testers, respectively, and evaluated as hybrids in two locations in 2018 and 2019 using an incomplete block design. Markers were derived from a Practical Haplotype Graph built from the founder whole genome assemblies and genotype-by-sequencing and exome capture-based sequencing of the population. Genetic mapping utilizing an update to R/qtl2 revealed differing profiles of significant loci for both traits between 635 of the DH lines and two sets of 570 and 471 derived hybrids. Genomic prediction was used to test the feasibility of predicting hybrid phenotypes based on the per se data. Predictive abilities were highest on direct models trained using the data they would predict (0.55 to 0.63), and indirect models trained using per se data to predict hybrid traits had slightly lower predictive abilities (0.49 to 0.55). Overall, this finding is consistent with the overlapping and non-overlapping significant QTL found within the per se and hybrid populations and suggests that selections for phenology traits can be made effectively on doubled haploid lines before hybrid data is available.

Seed increase through manual pollination is a critical part of maize breeding and genetics research to advance generations in breeding programs, to create desired research crosses, and produce hybrid seed for trials. Pollination in the field and in controlled environments relies on the availability of high‐quality pollen at the time that recipient silks are receptive. Generally, pollinations are made by capturing pollen from the tassel in a paper pollinating bag placed on the tassels one day prior to pollination and newly released pollen is then transferred to silks on the target plant. In the field, maize pollen is only viable for one to four hours following dehiscence and the rate of desiccation is influenced by environmental conditions. We have developed a method which increases the lifespan of pollen and allows pollen from a single tassel to be used to pollinate many ears by mixing fresh pollen with a dilutant that can be stored for multiple days. We identified characteristics of the size of suitable substrates and selected a PEEK‐based substrate for regular utilization. We evaluated pollen viability and empirically demonstrated the capability to store pollen up to 9 days when pollen is mixed with a PEEK substrate and stored at 6°C. The pollen storage method was used to make successful pollinations across 24 maize inbred lines tested and was generally equivalent to the standard manual pollination process. This method has the potential to increase the efficiency of breeding operations and may be useful in an array of genetic studies.

Seed increase through manual pollination is a critical part of maize breeding and genetics research to advance generations in breeding programs, to create desired research crosses, and produce hybrid seed for trials. Pollination in the field and in controlled environments relies on the availability of high-quality pollen at the time that recipient silks are receptive. Generally, pollinations are made by capturing pollen from the tassel in a paper pollinating bag placed on the tassels one day prior to pollination and newly released pollen is then transferred to silks on the target plant. In the field, maize pollen is only viable for one to four hours following dehiscence and the rate of desiccation is influenced by environmental conditions. We have developed a method which increases the lifespan of pollen and allows pollen from a single tassel to be used to pollinate many ears by mixing fresh pollen with a dilutant that can be stored for multiple days. We identified characteristics of the size of suitable substrates and selected a PEEK based substrate for regular utilization. We evaluated pollen viability and empirically demonstrated the capability to store pollen up to nine days when pollen is mixed with a PEEK substrate and stored at 6 degrees Celsius. The pollen storage method was used to make successful pollinations across 24 maize inbred lines tested and was generally equivalent to the standard manual pollination process. This method has the potential to increase the efficiency of breeding operations and may be useful in an array of genetic studies.

The Stiff Stalk heterotic pool is a foundation of US maize seed parent germplasm and has been heavily utilized by both public and private maize breeders since its inception in the 1930’s. Flowering time and plant height are critical characteristics for both inbred parents and their test crossed hybrid progeny. To study these traits, a six parent multiparent advanced generation intercross (MAGIC) population was developed including maize inbred lines B73, B84, PHB47 (B37 type), LH145 (B14 type), PHJ40 (novel early Stiff Stalk), and NKH8431 (B73/B14 type). A set of 779 doubled haploid lines were evaluated for flowering time and plant height in two field replicates in 2016 and 2017, and a subset of 689 and 561 doubled haploid lines were crossed to two testers, respectively, and evaluated as hybrids in two locations in 2018 and 2019 using an incomplete block design. Markers were derived from a Practical Haplotype Graph built from the founder whole genome assemblies and genotype-by-sequencing and exome capture-based sequencing of the population. Genetic mapping utilizing an update to R/qtl2 revealed differing profiles of significant loci for both traits between 636 of the DH lines and two sets of 571 and 472 derived hybrids. Genomic prediction was used to test the feasibility of predicting hybrid phenotypes based on the per se data. Predictive abilities were highest on direct models trained using the data they would predict (0.55 to 0.63), and indirect models trained using per se data to predict hybrid traits had slightly lower predictive abilities (0.49 to 0.55). Overall, this finding is consistent with the overlapping and non-overlapping significant QTL found within the per se and hybrid populations and suggests that selections for phenology traits can be made effectively on doubled haploid lines before hybrid data is available.Core IdeasA multi-parent advanced generation intercross (MAGIC) mapping population was developed from six founder Stiff Stalk maize inbreds with commercial relevance. Genetic mapping utilizing an update to R/qtl2 was demonstrated for flowering and plant height traits.Genetic mapping using maize inbred and hybrid information was compared and provided insight into trait expression in inbreds relative to heterotic testcross hybrids.