Elizabeth W. McCarthy’s research while affiliated with SUNY Cortland and other places

Allopolyploidy, a phenomenon prevalent in angiosperms involving hybridization and whole-genome duplication, results in species with multiple subgenomes, altering genome structure and gene expression, leading to novel phenotypes. Allopolyploids often experience unbalanced homeolog expression bias, the preferential expression of homeologs from one of the two progenitor genomes. To explore the consequences of allopolyploidy and unbalanced homeolog expression bias, we investigate global gene expression and the fate of homeologs in Nicotiana (Solanaceae). We focus on Nicotiana section Repandae, including three allotetraploid species, Nicotiana nudicaulis, N. repanda, and N. stocktonii, derived from diploid progenitors N. sylvestris and N. obtusifolia ∼4.3 Ma. We identify genes with differential expression and investigate expression of candidate genes for flower size variation. Our results show expression differences with the allopolyploids intermediate between the two progenitor species, with a slight bias toward N. obtusifolia. Moreover, we demonstrate unbalanced homeolog expression bias toward the N. obtusifolia subgenome across developmental stages in the allopolyploids, with a stronger bias in N. nudicaulis. In contrast, unbalanced homeolog expression bias shifts toward N. sylvestris for flower size genes in N. nudicaulis, showing that genes involved in particular phenotypes can display different patterns of unbalanced homeolog expression than the overall transcriptome. We also see differential expression of several known flower size genes across corolla developmental stages. Our results highlight the role of unbalanced homeolog expression bias in shaping the evolutionary trajectory of Nicotiana species and provide a foundation for future research into the ecological and evolutionary implications of allopolyploidy in flowering plants.

Homeologue expression bias occurs when one progenitor copy of a gene is expressed at a higher level than the other in allopolyploids. Morphological variation, including differences in flower colour, exists between natural and synthetic allopolyploids of Nicotiana tabacum and their progenitors. In this study, we use a comparative transcriptomic approach to investigate gene expression differences as well as homeologue bias in the flavonoid biosynthetic pathway (FBP) in these accessions. We do not observe reciprocal homeologue bias between dark and light pink allopolyploids, but the production of light pink flowers is correlated with high FLAVONOL SYNTHASE:DIHYDROFLAVONOL-4-REDUCTASE (FLS:DFR) ratio at 60% of anthesis length due to delayed activation of DFR in these accessions. We do find that natural allopolyploids have stronger homeologue bias than synthetic allopolyploids in both FBP genes and across the transcriptome. While there is no overall subgenome dominance, there is a bias towards expression of N. tomentosiformis homeologues in FBP genes; however, the magnitude of this bias is reduced in allopolyploids compared to the progenitors, suggesting that N. sylvestris homeologues play an active role in the development of flower colour in N. tabacum allopolyploids. In addition, synthetic allopolyploids tend to exhibit trans regulation of homeologues whereas natural allopolyploids often have evolved cis-regulatory differences between homeologues since their origin.

Floral organ size, especially the size of the corolla, plays an important role in plant reproduction by facilitating pollination efficiency. Previous studies have outlined a hypothesized organ size pathway. However, the expression and function of many of the genes in the pathway have only been investigated in model diploid species; therefore, it is unknown how these genes interact in polyploid species. Although correlations between ploidy and cell size have been shown in many systems, it is unclear whether there is a difference in cell size between naturally occurring and synthetic polyploids. To address these questions comparing floral organ size and cell size across ploidy, we use natural and synthetic polyploids of Nicotiana tabacum (Solanaceae) as well as their known diploid progenitors. We employ a comparative transcriptomics approach to perform analyses of differential gene expression, focusing on candidate genes that may be involved in floral organ size, both across developmental stages and across accessions. We see differential expression of several known floral organ candidate genes including ARF2, BIG BROTHER, and GASA/GAST1. Results from linear models show that ploidy, cell width, and cell number positively influence corolla tube circumference; however, the effect of cell width varies by ploidy, and diploids have a significantly steeper slope than both natural and synthetic polyploids. These results demonstrate that polyploids have wider cells and that polyploidy significantly increases corolla tube circumference.

Nicotiana tabacum L accessions vary in flower color from light pink to magenta. The differences in flower color are attributable to differences in anthocyanin content. To determine the genetic basis of flower color differences, we generated transcriptomes and quantified transcript levels of flavonoid biosynthetic genes in four N. tabacum accessions and their diploid progenitors. High expression ratios of the flavonol synthase (FLS) gene to dihydroflavonol 4-reductase (DFR) gene are found in light-pink flowers, suggesting that competition between the FLS and DFR enzymes for the same substrates may drive the flux of the flavonoid biosynthetic pathway toward producing flavonols at the expense of anthocyanins, resulting in light-pink flowers. The high FLS:DFR expression ratio appears to be attributable to DFR activation later in development in light-pink flowers.

Background Polyploidy has played a major role in angiosperm evolution. Previous studies have examined polyploid phenotypes in comparison to their extant progenitors, but not in context of predicted progenitor phenotypes at allopolyploid origin. In addition, differences in the trends of polyploid versus diploid evolution have not been investigated. We use ancestral character-state reconstructions to estimate progenitor phenotype at allopolyploid origin to determine patterns of polyploid evolution leading to morphology of the extant species. We also compare trends in diploid versus allopolyploid evolution to determine if polyploidy modifies floral evolutionary patterns. Results Predicting the ancestral phenotype of a nascent allopolyploid from reconstructions of diploid phenotypes at the time of polyploid formation generates different phenotype predictions than when extant diploid phenotypes are used, the outcome of which can alter conclusions about polyploid evolution; however, most analyses yield the same results. Using ancestral reconstructions of diploid floral phenotypes indicate that young polyploids evolve shorter, wider corolla tubes, but older polyploids and diploids do not show any detectable evolutionary trends. Lability of the traits examined (floral shape, corolla tube length, and corolla tube width) differs across young and older polyploids and diploids. Corolla length is more evolutionarily labile in older polyploids and diploids. Polyploids do not display unique suites of floral characters based on both morphological and color traits, but some suites of characters may be evolving together and seem to have arisen multiple times within Nicotiana, perhaps due to the influence of pollinators. Conclusions Young polyploids display different trends in floral evolution (shorter, wider corolla tubes, which may result in more generalist pollination) than older polyploids and diploids, suggesting that patterns of divergence are impacted by the early consequences of allopolyploidy, perhaps arising from genomic shock and/or subsequent genome stabilization associated with diploidization. Convergent evolution in floral morphology and color in Nicotiana can be consistent with pollinator preferences, suggesting that pollinators may have shaped floral evolution in Nicotiana. Electronic supplementary material The online version of this article (10.1186/s12870-019-1771-5) contains supplementary material, which is available to authorized users.