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Self-incompatibility

Self-incompatibility (SI) is a genetically controlled reproductive barrier in many s that prevents and fertilization by from genetically identical individuals, thereby promoting and maintaining within populations. This mechanism evolved independently multiple times across angiosperms and is estimated to occur in approximately 45% of flowering plant , serving as a key to avoid . At its core, SI relies on a multi-allelic S-locus that encodes recognition molecules in both and pistil tissues, enabling the plant to distinguish self from non-self pollen through specific molecular interactions. SI systems are broadly classified into two types based on the timing and genetic basis of pollen rejection: gametophytic self-incompatibility (GSI) and sporophytic self-incompatibility (SSI). In GSI, which predominates in families like (e.g., and ) and (e.g., many fruit trees), the incompatibility response is determined by the haploid of the itself; carrying an S-haplotype matching either of the pistil's two haplotypes fails to grow a beyond the upper . The molecular basis often involves S-RNase proteins secreted by the pistil, which degrade in incompatible , halting their growth. In contrast, SSI, common in (e.g., and ), is governed by the diploid of the pollen parent, with rejection occurring on the surface before tube emergence; this system features interaction between the pistil's S-receptor (SRK) and pollen's S-locus cysteine-rich protein (SCR), triggering a signaling that inhibits hydration and . A third, less common system in (e.g., poppies) involves rapid in incompatible via and disruption upon recognition by pistil PrsS proteins. The evolutionary and ecological significance of SI lies in its role as a self-recognition that enhances adaptability in heterogeneous environments, though it can break down through , , or environmental modifiers, leading to self-compatibility in some lineages and influencing crop breeding strategies. For instance, loss-of-function at the S-locus have repeatedly occurred, as seen in transitions from SI to self-compatibility in model systems like . Modern genomic tools, such as /, are now being applied to manipulate SI for hybrid seed production in crops like tomatoes and brassicas, underscoring its ongoing relevance in and .

Overview

Definition and Occurrence

Self-incompatibility (SI) is a genetic mechanism in angiosperms that prevents self-fertilization by rejecting from the same or genetically identical individuals, thereby promoting through . This process ensures that growth is inhibited or germination fails upon recognition of self- (incompatible pollen), while allowing fertilization by pollen from unrelated . SI operates primarily in hermaphroditic species, where both male and female reproductive organs are present in the same flower, countering the risk of inherent in such floral structures. SI occurs in approximately 45% of studied angiosperm species, making it a widespread reproductive strategy across the roughly 328,000 known species (as of 2024). It is particularly prevalent in families such as (e.g., petunias and potatoes), (e.g., apples and cherries), and (e.g., and ), where it enforces to maintain population-level . This distribution highlights SI's role as a key evolutionary adaptation in diverse ecosystems, from temperate orchards to agricultural fields. The historical discovery of traces back to key early observations in heterostylous plants, where first described the phenomenon in 1862 while studying the dimorphic flowers of . In these experiments, he noted the consistent failure of to produce seeds, contrasting with successful cross-pollination between floral morphs, which laid the groundwork for recognizing homomorphic SI in non-dimorphic species. These findings underscored SI as a physiological barrier distinct from structural adaptations, influencing subsequent research into its genetic basis.

Biological Importance

Self-incompatibility (SI) serves as a critical mechanism in many hermaphroditic flowering plants to promote by rejecting self- and often pollen from close relatives, thereby enhancing heterozygosity and within populations. This outcrossing enforcement avoids the accumulation of deleterious recessive alleles, mitigating and thereby increasing overall population fitness. For instance, in species like those in the family, SI maintains high allelic diversity at the S-locus, with over 50 distinct alleles reported in some populations, which supports robust genetic variation essential for long-term viability. Ecologically, SI influences plant mating systems by restricting gene flow to compatible mates, which can facilitate adaptation in heterogeneous or fragmented habitats where pollinator services may be limited. In such environments, the maintenance of S-allelic diversity ensures sufficient compatible partners, preventing mate limitation and promoting effective pollen transfer between populations. Furthermore, by preserving high levels of allelic polymorphism through balancing selection, SI contributes to reproductive isolation and speciation events, as divergent S-haplotypes can reduce interbreeding between emerging lineages. Despite these advantages, SI imposes fitness costs, including energetic investments in molecular recognition systems for pollen rejection and potential reductions in seed set when compatible pollen is scarce. These costs, such as lower reproductive output in small or isolated populations due to eroded S-allele richness, are generally offset by the long-term genetic benefits of reduced and enhanced adaptability.

Genetic Foundations

S-Locus Structure and Alleles

The self-incompatibility (SI) locus, commonly referred to as the S-locus, is a highly polymorphic genomic region that controls the recognition of self- in many flowering plants, ensuring by rejecting sharing the same S-haplotype. This multigenic locus consists of tightly linked encoding pistil-specific and -specific determinants, which function in a coordinated manner to mediate -pistil interactions. In gametophytic self-incompatibility (GSI) systems, prevalent in families such as and , the S-locus includes the S-RNase , which encodes a pistil-expressed responsible for rejection, and the S-locus F-box (SLF or SFB) , which encodes a -specific F-box protein involved in countering the RNase activity. In sporophytic self-incompatibility (SSI) systems, as seen in , the locus comprises the S-receptor kinase (SRK) in the pistil, a transmembrane receptor that perceives signals, and the S-cysteine-rich (SCR) or S-locus protein 11 (SP11) in , which acts as the for SRK activation. These linked determinants form a co-adapted complex, with the and pistil components evolving in concert to maintain specificity. The S-locus exhibits extraordinary allelic diversity, with natural populations typically harboring dozens to hundreds of distinct S-alleles, a level of polymorphism far exceeding that of other loci and maintained by balancing selection that favors rare alleles to promote outcrossing. This diversity arises from the functional necessity for unique recognition specificities, as each S-allele encodes distinct male and female determinants that interact in an allele-specific manner. In SSI, S-alleles display codominance, where the diploid sporophytic tissue (e.g., pollen exine) expresses both maternal and paternal alleles, leading to rejection if either matches the pistil's alleles. Conversely, in GSI, expression is gametophytic, with the haploid pollen's phenotype determined solely by its own S-allele, resulting in rejection only if it matches one of the pistil's two alleles. Such allelic variation ensures that compatible matings are frequent in diverse populations, with estimates indicating at least 45-50 alleles in some species like Oenothera organensis. The haplotype structure of the S-locus is characterized by strong suppression of recombination, creating large, non-recombining genomic islands that preserve the linkage between pistil and pollen determinants. This recombination suppression, often spanning hundreds of kilobases, is facilitated by structural features such as transposable element accumulations and inversions, which act as barriers to crossing-over and maintain haplotype integrity across generations. In Brassicaceae, for instance, the S-locus forms a supergene complex with tight linkage disequilibrium between SRK and SCR, encompassing repetitive sequences that further inhibit recombination. Similarly, in GSI systems like those in Prunus, the region around S-RNase and SLF shows reduced recombination rates, ensuring that deleterious recombinants— which could unlink co-adapted specificity—are rare. This haplotype organization underscores the S-locus as a classic example of a balanced polymorphic supergene in plant evolution.

Inheritance Patterns in GSI and SSI

In gametophytic self-incompatibility (GSI), the S-locus is inherited in a codominant manner, with the pollen's incompatibility determined directly by its own haploid S-allele rather than the diploid pollen parent. growth is arrested in the style if the pollen's S-allele matches either of the two S-alleles expressed in the diploid pistil, ensuring rejection of self- or related while permitting cross-compatible to fertilize. This pattern maintains high allelic diversity through negative frequency-dependent selection, as rarer S-alleles confer a transmission advantage. A classic example of GSI inheritance occurs in , where compatibility follows a straightforward matching rule. For instance, a pistil with genotype S_1S_2 rejects carrying S_1 or S_2 but accepts with non-matching alleles like S_3 or S_4. This can be illustrated by a compatibility matrix for selected S-alleles:
Pistil GenotypeCompatible Pollen (S_3)Compatible Pollen (S_4)Incompatible Pollen (S_1)Incompatible Pollen (S_2)
S_1S_2YesYesNoNo
S_3S_4YesYesYesYes
Such matrices demonstrate that half of the from a heterozygous parent (S_1S_2) is typically incompatible on a genetically identical pistil, reinforcing . In contrast, sporophytic self-incompatibility (SSI) inheritance involves the diploid genotype of the pollen parent dictating the pollen's , with rejection occurring on the surface if either parental S- matches either pistil S-allele. Unlike GSI, SSI features dominance hierarchies among S-alleles, where a dominant allele can suppress the expression of a recessive one in heterozygous pollen parents, altering compatibility outcomes. This dominance is often regulated at the level and contributes to complex interaction patterns. In species, such as B. oleracea, S-haplotypes are classified into dominance groups (e.g., class I dominant over class II), leading to hierarchical control. For example, pollen from an S_1S_2 parent (where S_1 is dominant) may express primarily the S_1 , rejecting on S_1-carrying pistils but potentially accepting on S_2-carrying ones depending on the . A simplified compatibility matrix for two dominance classes highlights this:
Pistil GenotypePollen from S_1 (Dominant)Pollen from S_2 (Recessive)Pollen from S_1S_2 (Heterozygote)
S_1S_3NoYesNo (S_1 dominant)
S_2S_3YesNoYes
These patterns ensure robust while allowing flexibility through dominance, as observed in crop breeding contexts.

Primary Self-Incompatibility Mechanisms

Gametophytic Self-Incompatibility (GSI)

Gametophytic self-incompatibility (GSI) is characterized by phenotypic control exerted by the haploid of the pollen grain, where pollen tube growth is arrested within the if the pollen's single S-allele matches either of the two S-alleles in the diploid maternal pistil tissue. This recognition ensures that only pollen with a novel S-allele can successfully fertilize the ovules, promoting and . Unlike sporophytic self-incompatibility, GSI lacks influence from the diploid of the pollen parent, with rejection determined solely by the pollen's own gametophytic identity. At the cellular level, incompatible pollen in GSI systems typically germinates on the but experiences inhibited tube elongation in the , often accompanied by tube swelling, irregular deposition of callose plugs, and eventual cessation of growth before reaching the . In some cases, particularly on stigmas, pollen hydration may fail prior to if incompatibility is detected early, though post-germination tube inhibition is more common. These events prevent self- from achieving fertilization while allowing compatible tubes to navigate the stylar transmitting tract unimpeded. GSI manifests prominently in families such as Solanaceae, where species like Petunia hybrida and Nicotiana alata exhibit stylar arrest of incompatible tubes in the upper third of the style, and Rosaceae, including Prunus dulcis (almond) and Malus domestica (apple), with similar inhibition observed in the stylar tissue. These examples highlight GSI's role in diverse herbaceous and woody perennials, where tube rejection ensures reproductive isolation. As the dominant self-incompatibility mechanism in many angiosperms, GSI occurs in 17–25 plant families, particularly among herbaceous species, where it contrasts with rarer sporophytic systems by enforcing haploid-based rejection without diploid dominance interactions. This prevalence underscores GSI's evolutionary success in preventing inbreeding across a broad taxonomic range.

Sporophytic Self-Incompatibility (SSI)

Sporophytic self-incompatibility (SSI) is a reproductive barrier in which the rejection of pollen occurs at the surface and is controlled by the diploid of the pollen parent, rather than the haploid of the pollen itself. In this system, the of the pollen is determined by the sporophytic tissues surrounding it during , particularly the tapetum in the anther, which deposits proteins onto the pollen coat that reflect the S-locus alleles of the diploid parent plant. As a result, pollen carrying matching S-alleles from the same or related individuals is recognized and rejected by the recipient , preventing self-fertilization and promoting . This mechanism contrasts with gametophytic self-incompatibility by acting early in the process based on the maternal and paternal contributions to the pollen parent's . At the cellular level, SSI typically blocks pollen hydration and germination on the stigma surface, halting pollen tube growth before it can penetrate the style. Incompatible pollen fails to imbibe water or initiate metabolic activity, leading to its desiccation and rejection without significant tissue damage to the stigma. This rapid response ensures efficient resource allocation to compatible pollen. SSI is prevalent in families such as (e.g., and species) and (e.g., sunflowers and daisies), where dry stigmas facilitate surface-based interactions. Dominance effects play a key role in SSI, where interactions between S-alleles in the pollen parent can modify the incompatibility . In heterozygous individuals (e.g., S1S2), one S- may dominate, masking the expression of the other in the pollen coat, such that the pollen behaves as if it carries only the dominant . This tissue-specific dominance, observed in the pollen but not always in the , influences rejection patterns and can lead to complex compatibility outcomes in crosses. Such allelic hierarchies contribute to the of high polymorphism at the S-locus.

Molecular Mechanisms of GSI

S-RNase Based System

The S-RNase-based system represents the predominant molecular mechanism underlying gametophytic self-incompatibility (GSI) in numerous angiosperm families, including Solanaceae and Rosaceae, where it enforces self/non-self recognition to prevent inbreeding. In this pathway, the pistil produces polymorphic S-RNase glycoproteins that are secreted into the stylar transmitting tissue, serving as the female specificity determinant. These S-RNases exhibit ribonuclease activity, which, upon uptake into the pollen tube, degrades ribosomal RNA (rRNA) in self-pollen tubes, thereby inhibiting protein synthesis and arresting tube growth within the style. This cytotoxic action is allele-specific, ensuring that only pollen sharing an S-haplotype with the pistil is rejected, while compatible pollen proceeds to fertilization. On the pollen side, the male specificity determinant consists of S-locus F-box (SLF) proteins in or S-haplotype-specific F-box (SFB) proteins in certain species, such as . These F-box proteins form part of SCF (Skp1-Cullin-F-box) E3 complexes that selectively target non-self S-RNases for polyubiquitination at specific residues and subsequent degradation by the 26S . This detoxification process neutralizes the inhibitory effect of non-self S-RNases, allowing compatible tubes to grow unimpeded. In contrast, self S-RNases evade ubiquitination due to allelic specificity conferred by hypervariable regions in both S-RNase and SLF/SFB proteins, remaining stable and active to block self-. Additionally, in some cases, undegraded self S-RNases can form condensates that disrupt the pollen tube and trigger , amplifying the rejection response. The in this system follows the general inhibition hypothesis, which posits that all stylar S-RNases initially inhibit growth indiscriminately upon entry, but pollen-expressed SLF/SFB proteins selectively degrade only non-self S-RNases, leaving self S-RNases to exert their cytotoxic effects. This model accounts for the collaborative non-self observed across alleles and explains phenomena like competitive interaction, where multiple non-self S-RNases can be detoxified simultaneously in heteroallelic . Allelic specificity arises from interactions between hypervariable domains (e.g., and regions in S-RNases) and corresponding motifs in SLF/SFB proteins, enabling precise matching. Seminal studies in hybrida () demonstrated this through identification of multiple SLF genes (up to 17 per ) that collectively handle non-self . In species, such as sweet cherry (Prunus avium) and almond (Prunus dulcis), the system relies on a single SFB gene per haplotype, which interacts with a general S-RNase inhibitor to modulate degradation, emphasizing a self-recognition variant of the general inhibition model. Here, SFB prevents the detoxification of matching self S-RNases, ensuring their persistence and inhibition. In contrast, species, including tomato relatives like Solanum chacoense, employ multiple divergent SLF genes that provide broad-spectrum non-self S-RNase degradation, with allelic specificity linked to specific SLF-S-RNase pairs. These differences highlight evolutionary adaptations within the S-RNase framework, yet both maintain the core cytotoxic mechanism via rRNA degradation. In gametophytic self-incompatibility (GSI), S-glycoproteins represent a class of non-RNase pistil-expressed proteins that mediate allele-specific recognition and rejection of self-pollen through signaling pathways rather than enzymatic degradation. These glycoproteins, secreted by the , interact directly with pollen tube receptors to initiate rapid physiological responses, including calcium influx that disrupts growth. Unlike degradative mechanisms, S-glycoproteins in these systems trigger non-cytotoxic signaling cascades that culminate in , ensuring precise self/non-self discrimination. A prominent example occurs in the Papaveraceae family, particularly in Papaver rhoeas (field poppy), where the pistil determinant PrsS is a small, secreted S-glycoprotein with multiple disulfide bonds that confers structural stability. Upon compatible allelic matching, PrsS binds to the pollen-specific transmembrane receptor PrpS, inducing receptor clustering and a rapid influx of cytosolic calcium ions into the pollen tube within seconds of contact. This calcium signal activates downstream mitogen-activated protein kinase (MAPK) cascades, notably the phosphorylation of a 56-kDa MAPK (p56), which leads to actin cytoskeleton depolymerization, cessation of tube tip growth, and eventual bursting of the pollen tube approximately 1-2 hours post-recognition. Specificity arises from allele-specific conformational changes in PrsS-PrpS interactions, preventing cross-reactivity with non-self alleles while avoiding broad toxicity. In the (grass) family, GSI operates via a two-locus system involving S and Z haplotypes, with related variants featuring (ZP) domain proteins on the pistil side and DUF247 domain proteins in pollen. The Z-locus encodes pistil-expressed ZP-domain glycoproteins that likely form multimers to recognize matching S-locus pollen proteins, triggering signaling events that inhibit penetration without cytotoxic enzymatic activity. Recent genomic analyses in species like perennial ryegrass () have identified conserved synteny between S and Z loci, with allelic diversity in ZP-domain sequences ensuring haplotype-specific binding and rejection, distinct from single-locus GSI in other families. This system highlights evolutionary convergence on glycoprotein-mediated signaling for promotion in wind-pollinated grasses.

Molecular Mechanisms of SSI

Brassica SRK-SCR System

The SRK-SCR system represents the archetypal molecular mechanism of sporophytic self-incompatibility (SSI) in the family, where recognition of self- occurs at the surface prior to . This system relies on allele-specific interactions between two key proteins encoded at the highly polymorphic S locus: the pistil determinant S-locus receptor (SRK) and the pollen determinant S-cysteine rich protein (SCR, also known as S-locus protein 11 or SP11). SRK functions as a transmembrane serine-threonine receptor predominantly expressed in the epidermal cells (papillae) of the , where it perceives pollen-derived signals. The SRK gene was first cloned from in , revealing its structure with an extracellular domain for ligand binding, a single transmembrane domain, and an intracellular domain responsible for . SCR serves as the pollen-specific ligand, a small, secreted cysteine-rich protein (approximately 75-85 amino acids) that accumulates in the pollen coat exine during maturation. Encoded by the tightly linked SCR/SP11 gene at the S locus, SCR is released upon pollen-stigma contact and binds specifically to the extracellular domain of the co-dominant SRK allele in an S-haplotype-specific manner. This interaction was demonstrated in seminal studies from 2001, which used recombinant proteins to show direct, allele-specific binding between SCR and the SRK ectodomain, confirming SCR as the male specificity determinant identified earlier through genomic mapping in B. oleracea and Brassica campestris (syn. B. rapa). Upon self-SCR binding, SRK undergoes dimerization and autophosphorylation at specific serine and threonine residues in its kinase domain, activating downstream phosphorylation cascades that transduce the rejection signal within the stigmatic cell. The activated SRK signaling pathway rapidly inhibits incompatible pollen behavior by preventing hydration and germination on the stigma surface, thereby blocking pollen tube emergence without affecting compatible pollen. This arrest occurs within minutes of pollination and involves actin cytoskeleton rearrangements and endocytosis in the stigmatic papillae, though the exact downstream effectors remain under investigation. Allele specificity in SRK-SCR recognition is governed by hypervariable regions (HV I-III) in the SRK extracellular domain and corresponding variable loops in SCR, which form complementary surfaces for precise lock-and-key interactions; structural analyses of SRK-SCR complexes, such as the S8 and S9 haplotypes, have revealed how sequence polymorphisms in these regions enable self/nonself discrimination while maintaining broad allelic diversity. Detailed functional studies of the SRK-SCR system have been conducted in B. oleracea, where natural SSI phenotypes were first genetically dissected, and in Brassica rapa, which provided crystal structures elucidating binding interfaces. Transgenic models in self-compatible Arabidopsis thaliana expressing matched Brassica SRK and SCR alleles have recapitulated SSI, confirming the sufficiency of these two genes for pollen rejection and enabling dissection of signaling components like the E3 ubiquitin ligase ARC1. These experimental systems highlight the system's evolutionary conservation across Brassicaceae, with over 100 characterized S haplotypes maintaining high polymorphism under balancing selection.

Other SSI Molecular Interactions

In the Convolvulaceae family, particularly in Ipomoea trifida, a close relative of sweet potato, sporophytic self-incompatibility (SSI) is controlled by a multiallelic S-locus that encodes SRK-like receptor kinases in the stigma and pollen-expressed ligands that facilitate specific recognition. These SRK-like kinases, expressed in the pistil, interact with pollen-derived ligands, including S-locus cysteine-rich (SCR)-like proteins that are anchored to the pollen surface via glycosylphosphatidylinositol (GPI) modifications, enabling haplotype-specific binding and activation of downstream rejection signals that inhibit pollen germination on the stigma surface. This system exhibits a linear dominance hierarchy among S-alleles, where dominant alleles can suppress recessive ones in heterozygous plants, contributing to varied incompatibility outcomes. In the Asteraceae family, SSI mechanisms show diversity beyond the canonical Brassica model, with evidence of conserved downstream signaling components such as E3 ubiquitin ligases. For instance, in Erigeron breviscapus, the EbARC1 gene encodes an E3 ubiquitin ligase that localizes to the nucleus and interacts with the exocyst subunit EbExo70A1, promoting its ubiquitination and degradation to enforce pollen rejection during self-pollination. Experimental overexpression of EbARC1 in the self-compatible Arabidopsis thaliana induced an SSI-like response, reducing self-pollen tube growth and seed set, highlighting its role in amplifying signaling cascades that block incompatible fertilization. Similarly, in Tolpis coronopifolia, genetic analyses reveal SSI controlled by the S-locus, where breakdowns in compatibility factors lead to partial self-compatibility, suggesting involvement of ubiquitin-mediated pathways in stabilizing rejection responses. Across non-Brassica SSI systems, common molecular themes include tightly linked receptor-ligand pairs at the S-locus, where pistil-expressed kinases recognize pollen-specific ligands to trigger allele-specific inhibition of or , often with high allelic diversity (e.g., exceeding 100 haplotypes in , though varying by family such as ~50 in ) to minimize . These interactions exhibit varying dominance relationships and cellular localizations—such as plasma anchoring in stigmas or GPI-linked presentation—allowing adaptation to diverse pressures, though the precise signaling effectors differ from the SRK-SCR paradigm. Recent studies (as of 2025) have further elucidated ubiquitination pathways in non-Brassica SSI, enhancing understanding of effector diversity.

Alternative Self-Incompatibility Systems

Two-Locus Gametophytic SI

Two-locus gametophytic self-incompatibility (GSI) is a genetic mechanism that controls pollen-pistil interactions in certain plant species, requiring matching alleles at two independent loci, designated S and Z, for rejection of self-pollen. In this system, unlike single-locus GSI, pollen tube growth is inhibited only when the pollen carries alleles that match those of the stigma at both the S and Z loci simultaneously. The S locus encodes specificity determinants expressed in both pollen and stigma tissues, while the Z locus contributes complementary recognition factors, ensuring that compatibility is determined by the combined haplotype. This dual-locus control results in a more complex recognition network, where is rejected only if its S matches one of the stigma's S alleles and its Z matches one of the stigma's Z alleles. For instance, in a diploid stigma with S1S2 Z3Z4, with S1Z3 would be incompatible, but with S1Z5 or S3Z3 would be compatible, as a single mismatch at either locus permits fertilization. This threshold requirement for dual matching enhances the precision of self-recognition while allowing broader cross-compatibility among heterozygous individuals. The two-locus GSI system is predominantly found in the grass family (Poaceae), particularly within the Pooideae subfamily, including species such as rye (Secale cereale), cocksfoot (Dactylis glomerata), and canary grass (Phalaris coerulescens). In Secale cereale, for example, the system has been extensively studied, revealing that self-incompatible populations maintain high allelic diversity at both loci, with pollen rejection occurring via failure of tube growth in the upper style when both loci match. This mechanism is widespread across self-incompatible grasses, contributing to their predominantly outcrossing breeding systems. The structural complexity of two-locus GSI substantially increases the number of possible allelic combinations compared to single-locus systems, with the total specificity groups equaling the product of the number of S and Z alleles, thereby promoting efficient and in natural . In species, this expanded combinatorial space can yield up to hundreds of unique S-Z specificities in a , far exceeding those in single-locus GSI, which supports robust avoidance of without overly restricting mate availability. Such intricacy has evolutionary implications for maintaining heterozygosity in wind-pollinated grasses.

Heteromorphic Self-Incompatibility

Heteromorphic self-incompatibility (HSI) is a form of self-incompatibility in where mating compatibility is tightly linked to distinct floral morphologies, ensuring by restricting growth from self- or intra-morph pollinations. This system integrates physical barriers from flower structure with genetic recognition mechanisms, promoting between different floral morphs. HSI is most commonly associated with , a polymorphism in style length and anther positioning that evolved independently in over 30 angiosperm families. The primary types of HSI are distyly and tristyly. Distyly involves two floral morphs: the long-styled (L- or pin) morph, with styles exceeding anther height, and the short-styled (S- or thrum) morph, with anthers positioned above the stigma. In this system, legitimate pollination occurs only between different morphs, as self-pollen or intra-morph pollen fails to fertilize due to incompatibility. A classic example is found in species of the genus Primula, such as Primula vulgaris, where distyly enforces outcrossing and maintains balanced morph ratios through negative frequency-dependent selection. Tristyly extends this to three morphs—long-, mid-, and short-styled—with corresponding anther levels at two heights, allowing compatible crosses only between dissimilar morphs in a diallelic manner. Lythrum salicaria exemplifies tristyly, where the complex arrangement further reduces self-fertilization opportunities. Genetic control of HSI is governed by a supergene at the S-locus, a tightly linked chromosomal region that orchestrates both morphological traits and incompatibility responses. In distylous systems like Primula, the S-morph is heterozygous (S/s), while the L-morph is homozygous recessive (s/s); the S-haplotype often includes a hemizygous region suppressing recombination to preserve linkage. Key genes within this supergene, such as CYP734A50 in Primula, regulate style length by modulating brassinosteroid signaling and cell elongation, while others like GLO2 influence anther positioning. Tristyly involves two linked loci, S and M, with epistatic interactions determining the three morphs and their compatibility. This supergene structure ensures co-inheritance of morphology and self-incompatibility, preventing the breakdown of the system through recombination. The mechanisms of HSI combine physical and genetic barriers to enforce outcrossing. Physically, reciprocal herkogamy— the precise reciprocal positioning of stigmas and anthers—facilitates pollinator-mediated cross-pollen transfer while minimizing self-pollen deposition on receptive surfaces. Genetically, heteromorphic self-incompatibility rejects self- or intra-morph pollen at the stigmatic or stylar level, often involving dimorphic pollen traits like size or surface sculpturing that trigger rejection responses. In Primula, for instance, the S-locus genes mediate these interactions, linking morphological dimorphism directly to pollen-style recognition. Overall, heterostyly in HSI reduces inbreeding depression and pollen wastage, enhancing reproductive success in polymorphic populations.

Specialized Self-Incompatibility Forms

Cryptic Self-Incompatibility (CSI)

Cryptic self-incompatibility () is a subtle form of self-incompatibility in which self- is not completely rejected but experiences reduced success relative to outcross , particularly under conditions of competition on the . This mechanism results in higher rates of fertilization by outcross when both self and compatible are present, while still permitting some self-fertilization when outcross is scarce. First described in the wallflower Cheiranthus cheiri, CSI manifests as a competitive disadvantage for self- during or growth, without overt inhibition. In CSI, the process typically involves prefertilization competition where outcross pollen outperforms self-pollen in reaching the ovules, often due to slower growth rates or lower germination efficiency of self-pollen tubes. Similarly, in Eucalyptus urophylla and E. grandis (), self-pollen tubes reach the base of the style significantly slower (120 hours versus 96 hours for outcross in E. urophylla), leading to reduced yields from selfing. These patterns suggest CSI operates through subtle physiological or molecular interactions that favor outcross pollen without requiring a full rejection response, as seen in classical gametophytic self-incompatibility systems. The implications of include promoting partial to enhance while providing reproductive assurance through limited selfing, which can buffer against pollinator scarcity or mate limitation. In populations exhibiting , such as certain Clarkia species, this leads to mixed mating systems where selfing rates increase under low activity but remain biased toward when loads are diverse. Overall, represents an evolutionary intermediate that fine-tunes breeding strategies in hermaphroditic plants, potentially evolving from or alongside stronger incompatibility mechanisms to balance and seed production.

Late-Acting Self-Incompatibility (LSI)

Late-acting self-incompatibility (LSI) is a reproductive barrier in plants where self-pollen tubes successfully reach and often penetrate the ovules, leading to fertilization, but subsequent seed development fails, resulting in abortion days after pollination. This post-zygotic mechanism contrasts with pre-zygotic forms of self-incompatibility by allowing initial syngamy before rejection, typically manifesting as endosperm breakdown or embryo degeneration. Potential underlying processes include genomic imprinting, where parent-of-origin-specific gene expression disrupts endosperm balance in self-fertilized seeds, or failure in endosperm cellularization due to imbalanced maternal-paternal genome contributions. In some cases, LSI mimics interspecific incompatibility responses, triggering programmed cell death or resource reallocation away from selfed seeds. Examples of LSI are documented in various plant families, with notable occurrences in , such as Cerastium arvense and Stellaria holostea, where self-pollen tubes penetrate ovules but yield no viable seeds, while cross-pollination succeeds. In these species, rejection happens 4–7 days post-pollination, often linked to early expressed through lethal recessives or imprinting defects in the . Similar patterns appear in other families like (Schima superba), where selfed ovules abort due to pollen tube cessation near ovules and subsequent failure, reducing seed set to less than 1%. LSI differs from cryptic self-incompatibility (CSI) in that it permits full initial fertilization success followed by complete seed rejection, whereas CSI involves selective pollen tube attrition at the ovule level, allowing partial self-fertility under mixed pollination. This late rejection in LSI may be adaptive in sparse populations, promoting outcrossing by aborting selfed progeny while conserving resources for potential compatible pollinations, potentially representing an ancestral outbreeding strategy in certain angiosperm lineages.

Self-Compatibility and Evolutionary Transitions

Mechanisms of Breakdown to Self-Compatibility

Self-incompatibility (SI) can break down through various genetic and molecular mechanisms, leading to self-compatibility (SC) that allows self-fertilization and often facilitates rapid population expansion in plants. These breakdowns are common evolutionary transitions, observed repeatedly across angiosperms, and typically involve disruptions in the recognition or response pathways that enforce rejection. In sporophytic SI (SSI) systems like those in , SC often arises from mutations in S-locus receptor kinase (SRK) or S-locus cysteine-rich (SCR) genes, while in gametophytic SI (GSI) systems such as those in and , alterations in S-RNase or pollen S-locus F-box (SLF/SFB) genes are prevalent. Loss-of-function mutations at the S-locus represent a primary mechanism of SI breakdown. In SSI, frameshift or nonsense mutations in SRK or SCR abolish pollen-stigma recognition; for instance, in , SC evolved via independent loss-of-function alleles in these genes approximately 1–1.5 million years ago, decaying entire S-haplogroups. Similarly, in GSI, deletions or point mutations in S-RNase genes prevent RNase activity and pollen tube inhibition, as seen in Pyrus serotina where a full S4-RNase deletion conferred SC, and in species where a single-nucleotide deletion at position 443 disrupted S-RNase function. These mutations are often recessive and fixed in self-compatible lineages, highlighting their role in irreversible shifts to SC. Genetic modifiers unlinked to the S-locus provide another key pathway for SI disruption without direct S-gene alterations. In Brassica rapa, the MOD locus mutation suppresses the self-incompatibility response, leading to SC. In GSI, modifiers like the MDF gene in Petunia axillaris reduce S13-RNase expression, while HT modifier mutations in Nicotiana alata lower pistil HT protein levels essential for RNase stabilization, both resulting in pollen tube growth. Recent studies in Arabidopsis lyrata identified an S1-specific modifier that interacts epistatically with homozygosity at the S-locus, producing SC progeny in crosses and challenging traditional views of mutation-only breakdowns; this modifier's action was confirmed in 1,503 progeny, where SC occurred exclusively in S1 homozygotes. Such modifiers likely act via shared pathways with pathogen defense, explaining their evolutionary lability. Polyploidy and S-locus duplications induce competitive interactions that erode SI, particularly in GSI systems. In tetraploid Nicotiana or Solanum lineages, duplicated S-alleles in diploid pollen tubes create non-self-like conditions, allowing tube penetration despite matching pistil alleles. This mechanism underlies SC in over 60 independent losses in Solanaceae and is amplified in polyploids where dominant nonfunctional alleles mask SI. In SSI, polyploidy similarly promotes SC by altering dominance hierarchies at the S-locus. These events underscore how genome duplication, common in crop domestication, accelerates transitions to SC for breeding advantages. Transgenic and experimental manipulations further illuminate breakdown mechanisms, often confirming natural pathways. Antisense suppression of S3-RNase in Petunia inflata or SLG in Brassica restores fertility by silencing recognition components, mirroring natural mutations. In Arabidopsis, transferring functional S-loci from SI relatives to SC A. thaliana re-establishes SI, proving that downstream factors remain intact. These insights, drawn from model systems, emphasize that SI breakdown is multifaceted, involving direct S-locus changes, indirect modifiers, and ploidy effects, with profound implications for plant evolution and agriculture.

Evolutionary Origins and Shifts

Self-incompatibility () in angiosperms is believed to have arisen independently multiple times, with phylogenetic analyses indicating at least 21 distinct evolutionary origins across flowering plants. These origins are often tied to the of pre-existing genetic elements involved in pathogen defense, such as receptor-like kinases (RLKs) and T2-type ribonucleases (RNases). In the family, the pistil-side determinant SRK evolved from ancestral RLKs that function in disease resistance signaling, through tandem gene duplications that adapted the recognition mechanism for rejection. Similarly, S-RNases in gametophytic SI systems derive from T2 RNases, which originally played roles in fungal defense and tissue remodeling, with monophyletic clades suggesting a single origin for S-RNase-based SI in core dating back approximately 110 million years. This repurposing of defense-related genes facilitated the linkage of -pistil recognition into tightly co-evolved S-loci, promoting while leveraging existing molecular machinery for specificity. The maintenance of SI polymorphism relies heavily on balancing selection, which preserves high allelic diversity at the S-locus to ensure effective . Negative favors rare S-alleles, as carrying common alleles faces less competition for compatible mates, thereby stabilizing dozens to hundreds of functional haplotypes within populations. This selective pressure counteracts and mutation, even through events like whole-genome duplications, by enforcing S-locus contractions or deletions that retain core recognition genes. In selfing or colonizing lineages, however, SI often breaks down due to reduced effective population sizes, where the reproductive assurance from self-fertilization outweighs outcrossing benefits, leading to fixation of loss-of-function mutations in S-genes. Transitions between and self-compatibility () exhibit directionality, with losses far more common than regains, though evidence suggests reversibility in certain clades through alternative mechanisms. In the , for instance, SI has been lost over 60 times, often via S-RNase inactivation, but regain via new SLF detoxifying systems or polyploidy-mediated restructuring has been inferred in some lineages. The costs of maintaining SI in small, fragmented populations—such as pollen discounting and reduced seed set from incompatible matings—drive these shifts, particularly during colonization events where mate availability is limited. Such evolutionary lability underscores SI's role as a conditionally adaptive trait, with breakdowns frequently referenced in mechanistic studies of evolution.

Distribution and Applications

Occurrence Across Plant Families

Self-incompatibility (SI) systems are distributed across approximately 40% of angiosperm lineages, encompassing at least 100 plant families, with distinct types prevailing in different major orders. Recent phylogenomic studies estimate SI occurs in approximately 39% of angiosperm species across more than 100 families. Gametophytic self-incompatibility (GSI) dominates in several core eudicot orders, including , , and , where it typically relies on an RNase-based involving S-RNase genes in the pistil that inhibit self-pollen tube growth. In , GSI is widespread in the family, such as tomatoes and nightshades, promoting through haploid pollen recognition. Similarly, (e.g., roses, apples) in and (e.g., blueberries) in exhibit this RNase-mediated GSI, contributing to high allelic diversity at the S-locus. Sporophytic self-incompatibility (SSI), in contrast, is prevalent in and , where pollen phenotype is determined by the diploid genotype, often leading to rejection at the stigma surface. The family (), including and mustards, exemplifies SSI through the S-locus with SRK and SCR/SP11 proteins that trigger rapid hydration inhibition. In , the (legumes like peas and beans) display various self-incompatibility systems, including gametophytic and late-acting types, though some subclades show variations or losses, maintaining in this economically vital group. Heteromorphic SI, characterized by floral polymorphisms like , occurs notably in Primulales, particularly the family. In species such as , distylous forms (short- and long-styled morphs) enforce legitimate crosses via sporophytic control, linking style length and anther position to prevent self-fertilization. Variations like (CSI) and late-acting self-incompatibility (LSI) appear in scattered families, such as those in . In (e.g., and ), CSI allows limited self-pollen success when cross-pollen is absent, while LSI acts post-fertilization in the , reducing self-seed viability and promoting in this order. SI is generally absent in basal angiosperms, including families like Amborellaceae and , where self-compatibility predominates without specialized recognition mechanisms.

Genetic Engineering and Research Advances

Genetic engineering approaches have targeted the S-locus in crops to facilitate production by disrupting self-incompatibility (SI) mechanisms. In , /Cas9-mediated editing of the S-locus gene SRK has produced stable self-compatible lines that simplify parental line maintenance and reduce breeding costs for F1 hybrids. Similarly, in Brassica napus, precise editing has generated novel self-incompatible variants by introducing specific mutations at the S-locus, enabling controlled for improved yield and uniformity in hybrids during the 2020s. Recent research advances have deepened understanding of SI signaling pathways through high-resolution techniques. In Papaver rhoeas, studies from 2023 have elucidated contrasting rejection mechanisms between Papaver's gametophytic SI and Brassica's sporophytic SI, highlighting rapid and disruption in incompatible tubes. A 2025 preprint revealed an integrated network of mitochondrially derived ROS and disruption of energy in incompatible Papaver tubes triggered by S-determinants. Additionally, sequencing of -stigma interactions has uncovered dynamic transcriptomic responses, including upregulated defense genes in incompatible pollen, providing insights into early rejection events. These advances support practical applications in and . Engineering SI breakdown in crops enhances rates, boosting vigor and resilience in species like . Furthermore, investigations into environmental influences reveal that rising temperatures under can destabilize SI, as elevated heat decreases RNase activity in Japanese , potentially accelerating self-compatibility transitions and altering .

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