Biology subjects

Neequaye, M.

Publications and source records attributed to Neequaye, M..

6 recordsLinked to original sources

bioRxiv2025

In-depth phenotyping reveals unexpected floral trait variation in Mimulus cardinalis across a range-wide latitudinal gradient

Background & AimsPollinators play a crucial role in the evolution and diversification of flowering plants. Intraspecific variation in floral traits occurs frequently and likely reflects variable biotic and abiotic selection pressures. The hummingbird-pollinated monkeyflower Mimulus cardinalis occurs across western North America, where geographic mosaics of selection have likely influenced its natural history, due to this wide species range. Processes driving early stages of divergence and speciation remain poorly understood, though theory predicts that trait divergence will be most likely at range edges. Within M. cardinalis, which is usually red-flowered and hummingbird-pollinated, two independent shifts to yellow flowers have occurred at the northern and southern range edges, including a population found on Cedros Island, off the coast of Baja California, Mexico. MethodsThis study characterises the local climatic variables of five accessions of M. cardinalis derived from locations across a latitudinal geographic gradient. Highly integrative methods in metabolomics and morphological analyses were used to characterise a suite of pollinator-relevant floral traits across the range of M. cardinalis species variation. We use transcriptomics and whole genome sequencing to study underlying genetic variation in two yellow-coloured, range-edge accessions. Key ResultsThis study uncovers high levels of variation in morphology, nectar properties, pigmentation and scent profile across geographically diverse Mimulus cardinalis accessions, in addition to profiling how these accessions are perceived by potential pollinators. We find high levels of phenotypic variation across M. cardinalis, particularly in the biochemistry of pigment and scent, where inflorescences that appear to be the same shade of red have completely different anthocyanin profiles. Floral trait-underlying genetic differences between different M. cardinalis lines were also investigated, revealing potential mechanisms underlying floral diversification. ConclusionsThis work highlights the importance of interplay between floral trait diversification, pollinator perception and climate. This work also highlights the importance of considering suites of floral traits in a quantitative fashion when studying intraspecific variation in relation to species range. This study also provides genetic insight into changes in these traits and provides many future directions to study the early stages of pollinator-mediated trait differentiation across a species range.

plant biology↗

bioRxiv2025

Redesigning petal shape, size, and color in soybean reveals unexpected phenotypes for floral organ development

Soybean (Glycine max) has not yet benefited from large-scale hybrid breeding efforts due to its small, self-fertilizing flowers that are difficult to emasculate, and limited attractiveness to pollinators. This study explores targeted floral trait engineering to enhance pollinator attraction, aiming to overcome barriers to soybean hybridization. We generated a high-resolution floral organ expression atlas and H3K4 trimethylation ChIP-Seq dataset to identify candidate genes involved in petal development, nectar sugar content, and petal pigmentation. Using CRISPR-based activation and repression systems, we modified the expression of AINTEGUMENTA (GmANT), BIGPETAL (GmBPE), and SUCROSE TRANSPORTER2 (GmSUC2). Contrary to expectations based on Arabidopsis homologs, transcriptional activation of GmANT_B and repression of GmBPE led to reduced, rather than increased, petal size, highlighting divergent regulatory mechanisms in soybean. Complementation of the W1 gene that controls petal pigmentation successfully converted white petals to purple, with preliminary evidence indicating that this color conversion may increase pollinator visitation. These results underscore the complexity of floral development in soybean and provide foundational tools and resources for future efforts to engineer reproductive traits for hybrid seed production.

plant biology↗

bioRxiv2025

Pollinator-relevant floral traits underlie bidirectional hybridisation in the orchid genus Gymnadenia

Reproductive isolation is mediated by ecological, phenotypic, and genetic barriers. Where such barriers are not upheld, hybridisation can occur, revealing insight about the formation of species. Here, we provide an integrative investigation of hybrid formation between the orchids Gymnadenia conopsea and G. rhellicani and potential isolating barriers in zones of overlap in the European Alps. We examine opportunities for hybridisation by quantifying hybrid frequency and pollinator assemblages of parent species. Next, we characterise floral phenotypes and genetic parentage of hybrids to examine how genetic inheritance of traits shapes ecological interactions, and thus fitness, of hybrids. We find that pollinator assemblages of parents overlap partially, consistent with observed rare hybrids (<1% of the population). Phenotypically, hybrids were intermediate across floral traits, consistent with all hybrids being identified as F1s, resulting from a primary hybridisation event between G. conopsea and G. rhellicani. No evidence of backcrossing or introgression was found, but direction of pollen transfer varied across sites. Additionally, hybrids were heterozygous for parental alleles at two loci impacting anthocyanin production, suggesting hybrids intermediate floral colour reflects inheritance of these alleles. Finally, we find little support that ploidy variation or hybrid seed infertility explains the absence of advanced generations of hybrids. Thus, we suggest that floral isolation represents an important isolating mechanism between G. conopsea and G. rhellicani in limiting the formation of uncommon, unfit hybrids and shaping species boundaries.

plant biology↗

bioRxiv2023

Do color polymorphisms reflect pollinator shifts?: Integrating floral traits as signals to a novel pollinator to understand species divergence

Understanding how evolution proceeds from molecules to organisms to interactions is critical for evolutionary biology and requires integrative studies that span biological levels. By linking phenotypic changes with associated genes and fitness implications, we gain insight on the repeatability of evolution and how adaptive walks move organisms between fitness peaks, which remains poorly understood at intermediate stages, e.g. potential fitness valleys. Floral trait evolution can proceed rapidly, confer reproductive isolation, and provides numerous examples of convergent evolution associated with pollinator guilds. Within the monkeyflowers (Mimulus sect. Erythranthe), intraspecific shifts to yellow flowers within red hummingbird-pollinated species have arisen at least twice and may represent incipient shifts between optima of pollinator adaptation. Here, we compare two natural yellow-flowered forms of M. cardinalis and M. verbenaceus to their red conspecifics in floral phenotypes, biochemistry, transcriptomic and genomic variation, and interactions with pollinators. We find convergent evolution in the adaptive walks of yellow morphs in both species, with consistent changes in traits of large effect (floral pigments and associated gene expression), which result in strong preference for yellow flowers by bumblebees. Shifts in additional traits (scent emissions and floral opening size) are also consistent with bee pollination, which may represent smaller-effect steps in an ongoing walk from hummingbird to bee pollination. By examining intraspecific variation representing incipient pollinator shifts in two related species, this work elucidates how adaptive walks proceed at early stages by highlighting how convergent large effect mutations (floral color) may drive pollinator attraction, preceding smaller effect changes controlling pollinator fit and reward access. This work integrates phenotypic, biochemical, genomic, and behavioral evidence to shed light on how repeated steps in adaptive walks to new fitness optima may impact reproductive isolation and incipient speciation via convergent evolution.

evolutionary biology↗

bioRxiv2020

The Effect of Introgression of an R2R3 MYB Transcription Factor on Sulphur Metabolism in Brassica oleracea

BackgroundA diet rich in cruciferous vegetables is reported to have beneficial health effects, partially mediated by 4-methylsulfinylbutyl glucosinolate, or glucoraphanin, which is predominantly found within broccoli (Brassica oleracea var italica). We describe the downstream effects on transcription and metabolism in broccoli following the introgression of a genetic variant of MYB28 into broccoli from a wild Brassica relative which has previously been associated with enhancement of glucoraphanin. ResultsWhole genome sequencing, RNA expression and metabolite analyses were used to characterise the consequences of the introgression of either one or two copies of a genetic variant of the MYB28 transcription factor into a commercial broccoli genetic background. The introgression of the variant of MYB28 resulted in enhanced expression of genes involved in primary sulphate assimilation, sulphur metabolism and aliphatic glucosinolate biosynthesis, and enhanced accumulation of 4-methylsulphinyl butyl glucosinolate in florets. Other changes in transcription that may be related to non-targeted introgression events are reported. There were no consistent effects upon sulphur metabolites pools, apart from methionine-derived glucosinolates. ConclusionThis study illustrates the downstream effects on transcription and metabolism of the introgression of a genetic variant of MYB28 from a wild species into a commercial broccoli genotype.

plant biology↗

bioRxiv2020

CRISPR-Cas9-mediated editing of myb28 impairs glucoraphanin accumulation of Brassica oleracea in the field

We sought to quantify the role of MYB28 in the regulation of aliphatic glucosinolate biosynthesis and associated sulphur metabolism in field-grown B. oleracea with the use of CRISPR-Cas9-mediated gene editing technology. We describe the first characterised myb28 knockout mutant in B. oleracea, and the first UK field trial of CRISPR-Cas9-mediated gene edited plants under the European Court of Justice interpretation of the 2001/18 EU GMO directive. We report that knocking-out myb28 results in downregulation of aliphatic glucosinolate biosynthesis genes and reduction in accumulation of the methionine-derived glucosinolate, glucoraphanin, in leaves and florets of field-grown myb28 mutant broccoli plants. There were no significant changes to the accumulation of sulphate, S-methyl cysteine sulfoxide and indole glucosinolate in leaf and floret tissues.

plant biology↗