In a groundbreaking study published in Nature, researchers have unveiled how hidden layers of genetic variation, known as cryptic cis-regulatory alleles, drive complex phenotypic changes in plants through a sophisticated hierarchical epistatic network. Using tomato inflorescence architecture as a model, the study dissects the intricate gene regulatory interactions that govern plant morphology, revealing new insights into how evolutionary potential is encoded and unleashed within species. This work not only harnesses natural genetic diversity but also meticulously engineers novel allelic combinations to map the detailed genotype–phenotype landscape with unprecedented resolution.
The authors focused on a four-gene network involving PLT and SEP transcription factor gene families that regulate the branching and development of tomato inflorescences. Through extensive combinatorial genotyping and phenotyping of thousands of inflorescences derived from engineered cryptic cis-regulatory variants, they uncovered that mutations within this network typically interact in a multiplicative manner. Strikingly, paralogous gene pairs—genes derived from duplication events within a species—show even stronger than multiplicative, so-called super-multiplicative, synergistic interactions. This finding underlines a fundamental role for paralogue redundancy and divergence in shaping the evolutionary dynamics of developmental traits.
Beyond simple multiplication, the study discovered a dose-dependent masking phenomenon between paralogue pairs. Specifically, mutations in one paralogue pair systematically diminish the phenotypic impact of mutations in the other pair, pointing to a complex layering of epistatic modulation that buffers or amplifies trait variability. This nuanced hierarchical interaction enriches our understanding of how genetic networks maintain developmental stability yet remain poised for significant phenotypic innovations when cryptic variants converge.
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Pan-genome sequencing across tomato species and their wild relatives provided the raw variation underpinning this analysis, highlighting the rich diversity of cis-regulatory landscapes molded by structural variations—ranging from small indels to large genomic duplications and deletions. These dynamic genomic rearrangements modulate gene expression dosage and network architecture, driving quantitative variation in plant development. Importantly, turnover in paralogue content between species reshapes network buffering capacity, thus potentiating phenotypic changes that may be “canalized” or otherwise hidden within stable developmental programs.
The research team constructed a mechanistic yet phenomenological model that situates the PLT–SEP regulatory network within a hierarchical framework of genetic interactions. The model captures classical synergistic epistasis observed within paralogue pairs and integrates these through a multilinear interaction between the pairs, ultimately governed by an exponential mapping to predict branching outcomes. This conceptual model bridges molecular gene regulatory logic with phenotypic outputs measured at the organ level, offering a powerful abstraction that informs both evolutionary biology and applied plant breeding.
Intriguingly, their findings provide much-needed empirical support for the multilinear epistasis model, which theorists have long proposed but has rarely been substantiated in natural systems. The model treats gene regulatory interactions as a spectrum of continuous allelic strengths rather than simplistic binary states, better reflecting biological complexity. This continuous framework explains how paralogue redundancy and divergence coordinate to simultaneously expand and constrain phenotypic space, setting the stage for nuanced evolutionary trajectories.
Comparative genomics revealed contrasting network architectures between Solanaceae—tomato’s family—and Brassicaceae, such as Arabidopsis, where certain paralogues like J2 and EJ2 MADS-box genes have been lost. This absence likely contributes to constrained branching phenotypes in the Brassicaceae lineage, whereas the presence and variation of these genes in Solanaceae enable the accumulation of cryptic variation that fuels morphological diversification. Such differences illuminate how taxon-specific gene content shapes regulatory network evolvability across evolutionary timescales.
The study’s approach—engineering a dense matrix of genotypic combinations within an isogenic background—was crucial for unraveling the quantitative form and character of epistasis in this complex network. The resulting genotype–phenotype surface maps provide a visual and mathematical representation of how mutations interact within and between paralogue families to influence inflorescence branching. This platform also offers a blueprint for interpreting the enigmatic epistatic effects frequently observed but difficult to detect in natural populations.
Beyond evolutionary insights, this framework holds transformative potential for crop engineering. Understanding how specific allelic combinations and their epistatic relationships generate equivalent phenotypic outcomes allows precise “tuning” of gene networks via targeted genome editing. Such strategies could optimize agronomic traits while minimizing negative pleiotropic effects and circumventing the genetic constraints imposed by naturally occurring allele combinations. This precision agriculture approach promises to accelerate breeding programs by exploiting network-aware epistatic landscapes.
The authors highlight that fully harnessing these opportunities will require scaling analyses to larger and more intricate gene networks. Additionally, exploring how species-specific complements of paralogous genes modulate hierarchical epistasis will deepen our comprehension of developmental and physiological adaptations across the plant kingdom. This calls for integrative efforts combining pan-genomics, functional genetics, and computational modeling to chart the genotype-to-phenotype map in three-dimensional complexity.
This transformative study thereby marks a significant leap in decoding the hidden plasticity within gene regulatory networks that underpin plant development and evolution. By leveraging cryptic variation and illuminating multilayered genetic interactions, it sets a new paradigm for understanding the genetic basis of morphological innovation. The implications span basic evolutionary theory, functional genomics, and practical agriculture, making hierarchical epistasis a central concept in modern biology’s quest to predict and manipulate phenotypic outcomes.
In summary, the investigation of cryptic regulatory variants within the PLT–SEP gene network showcases the elegance and complexity of evolutionary genetic architecture. It illuminates how gene duplication, structural genomic variation, and multilayered epistasis synergistically shape plant morphology and its evolvability. As researchers continue to unravel these intricate interactions, a future where phenotypes are predictably engineered by navigating sophisticated genotype landscapes seems increasingly attainable.
Subject of Research: Genetic architecture and epistasis in plant developmental networks, focusing on tomato inflorescence branching.
Article Title: Cryptic variation fuels plant phenotypic change through hierarchical epistasis.
Article References:
Zebell, S.G., Martí-Gómez, C., Fitzgerald, B. et al. Cryptic variation fuels plant phenotypic change through hierarchical epistasis. Nature (2025). https://doi.org/10.1038/s41586-025-09243-0
Image Credits: AI Generated
Tags: combinatorial genotyping in plantscryptic cis-regulatory allelesdose-dependent gene interaction phenomenaevolutionary potential in plantsgene regulatory interactionshierarchical epistatic networksmapping genotype-phenotype landscapesnatural genetic diversity in plantsparalogous gene pairs in evolutionplant phenotypic variationsuper-multiplicative gene interactionstomato inflorescence architecture