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Sex dimorphism and gene expression were studied in developing catkins in 159 F2 individuals from the bioenergy crop Salix purpurea, and potential mechanisms and pathways for regulating sex development were explored. Differential expression, eQTL, bisulfite sequencing, and network analysis were used to characterize sex dimorphism, detect candidate master regulator genes, and identify pathways through which the sex determination region (SDR) may mediate sex dimorphism. Eleven genes are presented as candidates for master regulators of sex, supported by gene expression and network analyses. These include genes putatively involved in hormone signaling, epigenetic modification, and regulation of transcription. eQTL analysis revealed a suite of transcription factors and genes involved in secondary metabolism and floral development that were predicted to be under direct control of the sex determination region. Furthermore, data from bisulfite sequencing and small RNA sequencing revealed strong differences in expression between males and females that would implicate both of these processes in sex dimorphism pathways. These data indicate that the mechanism of sex determination in Salix purpurea is likely different from that observed in the related genus Populus. This further demonstrates the dynamic nature of SDRs in plants, which involves a multitude of mechanisms of sex determination and a high rate of turnover.

Genetic improvement through breeding is one of the key approaches to increasing biomass supply. This paper documents the breeding progress to date for four perennial biomass crops (PBCs) that have high output-input energy ratios: namely Panicum virgatum (switchgrass), species of the genera Miscanthus (miscanthus), Salix (willow) and Populus (poplar). For each crop, we report on the size of germplasm collections, the efforts to date to phenotype and genotype, the diversity available for breeding and on the scale of breeding work as indicated by number of attempted crosses. We also report on the development of faster and more precise breeding using molecular breeding techniques. Poplar is the model tree for genetic studies and is furthest ahead in terms of biological knowledge and genetic resources. Linkage maps, transgenesis and genome editing methods are now being used in commercially focused poplar breeding. These are in development in switchgrass, miscanthus and willow generating large genetic and phenotypic data sets requiring concomitant efforts in informatics to create summaries that can be accessed and used by practical breeders. Cultivars of switchgrass and miscanthus can be seed-based synthetic populations, semihybrids or clones. Willow and poplar cultivars are commercially deployed as clones. At local and regional level, the most advanced cultivars in each crop are at technology readiness levels which could be scaled to planting rates of thousands of hectares per year in about 5 years with existing commercial developers. Investment in further development of better cultivars is subject to current market failure and the long breeding cycles. We conclude that sustained public investment in breeding plays a key role in delivering future mass-scale deployment of PBCs.

Genetic improvement through breeding is one of the key approaches to increasing biomass supply. This paper documents the breeding progress to date for four perennial biomass crops (PBCs) that have high output-input energy ratios: namely Panicum virgatum (switchgrass), species of the genera Miscanthus (miscanthus), Salix (willow) and Populus (poplar). For each crop, we report on the size of germplasm collections, the efforts to date to phenotype and genotype, the diversity available for breeding and on the scale of breeding work as indicated by number of attempted crosses. We also report on the development of faster and more precise breeding using molecular breeding techniques. Poplar is the model tree for genetic studies and is furthest ahead in terms of biological knowledge and genetic resources. Linkage maps, transgenesis and genome editing methods are now being used in commercially focused poplar breeding. These are in development in switchgrass, miscanthus and willow generating large genetic and phenotypic data sets requiring concomitant efforts in informatics to create summaries that can be accessed and used by practical breeders. Cultivars of switchgrass and miscanthus can be seed-based synthetic populations, semihybrids or clones. Willow and poplar cultivars are commercially deployed as clones. At local and regional level, the most advanced cultivars in each crop are at technology readiness levels which could be scaled to planting rates of thousands of hectares per year in about 5 years with existing commercial developers. Investment in further development of better cultivars is subject to current market failure and the long breeding cycles. We conclude that sustained public investment in breeding plays a key role in delivering future mass-scale deployment of PBCs.

Dioecy has evolved numerous times in plants, but heteromorphic sex chromosomes are apparently rare. Sex determination has been studied in multiple Salix and Populus (Salicaceae) species, and P. trichocarpa has an XY sex determination system on chromosome 19, while S. suchowensis and S. viminalis have a ZW system on chromosome 15. Here we use whole genome sequencing coupled with quantitative trait locus mapping and a genome-wide association study to characterize the genomic composition of the non-recombining portion of the sex determination region. We demonstrate that Salix purpurea also has a ZW system on chromosome 15. The sex determination region has reduced recombination, high structural polymorphism, an abundance of transposable elements, and contains genes that are involved in sex expression in other plants. We also show that chromosome 19 contains sex-associated markers in this S. purpurea assembly, along with other autosomes. This raises the intriguing possibility of a translocation of the sex determination region within the Salicaceae lineage, suggesting a common evolutionary origin of the Populus and Salix sex determination loci.

The first step in trait introgression is to identify and assess novel sources of variation. For shrub willow (Salix) breeders, there is an abundance of understudied species within a genus that readily hybridizes. Breeding targets in shrub willow center on traits contributing to biomass yield for bioenergy. These include stem biomass, insect and pathogen resistance, and leaf architecture traits. More specifically, breeding for durable resistance to willow leaf rust (Melampsora spp.) is of particular importance as the pathogen can significantly reduce biomass yields in commercial production. The Salix F1 hybrid common parent population (Salix F1 HCP) was created to characterize the variation among eight species‐hybrid families and map QTL for targeted traits. A female and male S. purpurea were used as common parents in crosses made to male S. suchowensis, S. viminalis, S. koriyanagi, and S. udensis and female S. viminalis, S. integra, S. suchowensis to produce eight families that were planted in field trials at Cornell AgriTech in Geneva, NY and phenotyped. Using 16 previously described parental backcross linkage maps and two newly generated S. purpurea consensus maps, we identified 215 QTL across all eight families and in every parent. These included 15 leaf rust severity, 61 herbivory, 65 leaf architecture, and 74 yield component QTL, resulting in 50 unique overlapping regions within the population. These genetic loci serve as an important foundation for future shrub willow breeding, and each interspecific family was identified as a novel source of useful alleles for trait introgression into high yielding cultivars. [ABSTRACT FROM AUTHOR]

Cannabinoid hemp is a dioecious crop where pistillate plants are selectively grown to maximize cannabinoid yields. Errant pollination can reduce pistillate flower yields, cannabinoid concentrations, and lead to unmarketable flowers due to the presence of undesirable seeds. We compared pollen sensitivity and agronomic performance of diploid (2n = 2x = 20 chromosomes) and triploid (2n = 3x = 30 chromosomes) cannabinoid hemp in open-field conditions. The high-CBD variety Lifter and high-CBG variety White CBG were evaluated in both their diploid and triploid counterparts in fields with and without pollen. Trials were conducted during the 2021 and 2022 field seasons in Kentucky, New York, and North Carolina (n = 6 site years). Triploids produced taller and wider plants than diploids; however, this did not result in yield differences in the pollen-free environment. In the presence of pollen, triploid Lifter and White CBG produced 87% and 77% fewer seeds than their diploid counterparts, respectively. Increased seed production in diploids also resulted in a significant reduction of seed-free biomass and cannabinoid concentrations compared to triploids. In the absence of pollen, we did not find any appreciable differences in seed-free biomass or cannabinoid concentrations between triploids and diploids. Though not completely pollen-insensitive, triploidy is an effective means to reduce seed production and improve yields and quality in cannabinoid hemp in the presence of pollen. [ABSTRACT FROM AUTHOR]

Woody perennial angiosperms (i.e., hardwood trees) are polyphyletic in origin and occur in most angiosperm orders. Despite their independent origins, hardwoods have shared physiological, anatomical, and life history traits distinct from their herbaceous relatives. New high-throughput DNA sequencing platforms have provided access to numerous woody plant genomes beyond the early reference genomes of Populus and Eucalyptus, references that now include willow and oak, with pecan and chestnut soon to follow. Genomic studies within these diverse and undomesticated species have successfully linked genes to ecological, physiological, and developmental traits directly. Moreover, comparative genomic approaches are providing insights into speciation events while large-scale DNA resequencing of native collections is identifying population-level genetic diversity responsible for variation in key woody plant biology across and within species. Current research is focused on developing genomic prediction models for breeding, defining speciation and local adaptation, detecting and characterizing somatic mutations, revealing the mechanisms of gender determination and flowering, and application of systems biology approaches to model complex regulatory networks underlying quantitative traits. Emerging technologies such as single-molecule, long-read sequencing is being employed as additional woody plant species, and genotypes within species, are sequenced, thus enabling a comparative ("evo-devo") approach to understanding the unique biology of large woody plants. Resource availability, current genomic and genetic applications, new discoveries and predicted future developments are illustrated and discussed for poplar, eucalyptus, willow, oak, chestnut, and pecan.

Summary: The Salicaceae, including Populus and Salix, are dioecious perennials that utilize different sex determination systems. This family provides a useful system to better understand the evolution of dioecy and sex chromosomes.Here, a rare monoecious genotype of Salix purpurea, 94003, was self‐ and cross‐pollinated and progeny sex ratios were used to test hypotheses on possible mechanisms of sex determination. To delimit genomic regions associated with monoecious expression, the 94003 genome sequence was assembled and DNA‐ and RNA‐Seq of progeny inflorescences was performed.Based on alignments of progeny shotgun DNA sequences to the haplotype‐resolved monoecious 94003 genome assembly and reference male and female genomes, a 1.15 Mb sex‐linked region on Chr15W was confirmed to be absent in monecious plants. Inheritance of this structural variation is responsible for the loss of a male‐suppressing function in what would otherwise be genetic females (ZW), resulting in monoecy (ZWH or WWH), or lethality, if homozygous (WHWH).We present a refined, two‐gene sex determination model for Salix purpurea, mediated by ARR17 and GATA15 that is different from the single‐gene ARR17‐mediated system in the related genus Populus. [ABSTRACT FROM AUTHOR]