Competition affects mixed-mating strategies by limiting available abiotic or biotic resources such as nutrients, water, space, or pollinators. Cleistogamous species produce closed (cleistogamous, CL), obligately selfed, simultaneously with open (chasmogamous, CH), potentially outcrossed flowers. The effects of intraspecific competition on fitness and cleistogamy variation can range from limiting the production of costly CH flowers because of resource limitation, to favouring CH production because of fitness advantages of outcrossed, CH offspring. Moreover, the effects of competition can be altered when it co-occurs with other environmental variations. We grew plants from seven populations of the ruderal Lamium amplexicaule, originating from different climates and habitats, in a common garden experiment combining drought, interspecific competition, and seasonal variation. All these parameters have been shown to influence the degree of cleistogamy in the species on their own. In spring, competition and drought negatively impacted fitness, but the CL proportion only increased when plants were exposed to both treatments combined. We did not observe the same results in autumn, which can be due to non-adaptive phenotypic variation, or to differences in soil compactness between seasons. The observed responses are largely due to phenotypic plasticity, but we also observed phenotypic differentiation between populations for morphological, phenological, and cleistogamy traits, pointing to the existence of different ecotypes. Our data do not support the hypothesis that CL proportion should decrease when resources are scarce, as plants with reduced growth had relatively low CL proportions. We propose that variation in cleistogamy could be an adaptation to pollinator abundance, or to environment-dependent fitness differences between offspring of selfed and outcrossed seeds, two hypotheses worth further investigation. This opens exciting new possibilities for the study of the maintenance of mixed-mating systems using cleistogamous species as models that combine the effects of inbreeding and reproductive costs.

Plants display various forms of phenotypic plasticity in anticipation of changing conditions, many of which are influenced by information obtained from neighbouring plants. Here, we tested the hypothesis that cleistogamic Lamium amplexicaule plants can adaptively modify production of chasmogamous (CH) and cleistogamous (CL) flowers based on the perception of conspecific neighbours. The production and proportion of CH and CL flowers was examined in individual L. amplexicaule grown at varying densities or treated with root leachates from plants grown at different densities. When growing at high density or treated with root leachates from high-density pots, L. amplexicaule increased production of more expensive, potentially outcrossing CH flowers. In contrast, single plants or plants treated with root leachates from empty pots or single-source plants predominantly developed cheaper, self-pollinated CL flowers. The results demonstrate a novel root-based neighbour-perception modality that enables plants to adaptively adjust production of CH and CL flowers in response to the presence of potential reproductive partners. Further research is needed to explore the broader ecological implications of this novel interplant cueing on reproductive bet-hedging and plasticity in natural settings, as well as to identify the involved cues and their mode of operation.

Lamium amplexicaule L. (Family: Lamiaceae) is a cosmopolitan weed whose eradication is challenging. The phenoplasticity of this species is related to its heteroblastic inflorescence, which has not received adequate research worldwide in its morphological and genetic aspects. This inflorescence hosts two flower types, a cleistogamous (CL: closed flower) and a chasmogamous (CH: opened flower). This species subjected to detailed investigation is a model species to clarify: (1) the existence of the CL and CH flowers in relation to the time and individual plants. (2) the predominant flower morphs in Egypt. (3) the morphological and genetic variability between these morphs. Among the novel data retrieved from this work is the Presence of this species in three distinct morphs coexisting during winter. These morphs showed remarkable phenoplasticity, particularly in flower organs. Significant differences were observed between the three morphs in pollen fertility, nutlets productivity and sculpture, flowering time, and seed viability. These differences were extended to the genetic profile of these three morphs assessed by the inter simple sequence repeats (ISSRs) and start codon targeted (SCoT). This work highlights the urgent need to study the heteroblastic inflorescence of crop weeds to facilitate its eradication.

OBJECTIVE: Many angiosperms exhibit cleistogamy, the production of both cleistogamous flowers (CL), which remain closed and obligately self-pollinated, and chasmogamous flowers (CH), which are potentially open-pollinated. The CH proportion can be plastic. Plasticity is adaptive if environmental changes can be reliably assessed and responded to with an appropriate phenotype and if plastic genotypes have higher fitness in variable environments than non-plastic ones.
METHODS: We studied the plastic response of four natural populations from northern and southern France of an annual cleistogamous plant, Lamium amplexicaule, to predictable seasonal variation. Plants were grown in a semi-controlled environment in spring and in autumn. We assessed the variation in flower number, phenology and cleistogamy-related traits, which were all plastic with respect to season. The CH proportion was higher in spring than in autumn in all four populations.
RESULTS: We showed significant stabilizing selection for cleistogamy traits, with higher optimal CH proportions and more pronounced stabilizing selection in spring than in autumn. Observed CH proportions were close to the predicted optimal CH proportions in each season except in autumn for southern populations, which do not experience the autumnal growing season in nature.
CONCLUSIONS: These results are consistent with adaptive plasticity across seasons of cleistogamy in L. amplexicaule.We propose that adaptive plasticity of cleistogamy could be driven by pollination environment variation, with CL flowers providing reproductive assurance when pollinators are scarce and CH flowers reducing the inbreeding depression in offspring when pollinators are abundant.

OBJECTIVE: Lamium amplexicaule is a cleistogamous plant that produces both closed flowers (obligately self-pollinated) and open flowers (potentially outcrossed). The conditions for the maintenance of such a mating system depend on the outcrossing rate of the open flowers, which can be estimated using neutral microsatellite markers. •
RESULTS: Forty primer pairs corresponding to microsatellite motifs obtained by coupling multiplex microsatellite enrichment and next-generation sequencing were tested. Thirteen primers amplified with satisfying results. The polymorphism of these markers was studied in four French populations. Allele number varied from one to eight per locus and per population. Heterozygosity levels were significantly lower than those expected under Hardy-Weinberg equilibrium. •
CONCLUSIONS: Our results are consistent with a partial self-fertilization pattern. These markers will be used to estimate the outcrossing rate as well as population differentiation in L. amplexicaule.