Cleistogamous Flowers Research Articles

Selective forces on the maintenance of outcrossing in an almost exclusively cleistogamous violet species.

Cleistogamous species constitute interesting study systems to resolve the longstanding question of how outcrossing is maintained given that seed production is ensured through selfing. In this work, we investigate the selective forces that allow the persistence of producing self-pollinated cleistogamous (CL) and chasmogamous (CH) flowers in Viola jaubertiana Marès & Vigin. We monitored three populations at different elevation for two years, and studied the flowering phenology and the relative contribution of each flower morph to parental fitness. We tested whether allocation to CH and CL flowers differed across populations and if it covaried with herbivory and water stress conditions. We also performed hand-pollination and bagging experiments in CH flowers to estimate inbreeding depression and heterosis. The CH flowers open in winter under unfavorable conditions for pollination, show high pollen limitation and no-delayed selfing, and thus produce a low amount of seeds. Conversely, CL flowers appear in early spring, are physiologically cheaper to produce (i.e., dry weight is 3.4 times lower than that of CH flowers), and yield approximately 100 times more seeds than CH flowers. The CH flowers were favored under water stress and low herbivory. Crosses between populations showed up to 25% greater fitness than those within populations. Despite the great pollen limitation in CH flowers, we suggest that the interaction among different environmental determinants and heterosis are probably sufficient forces to maintain chasmogamy in this long-lived species, reducing deleterious fixed mutations in the selfed lines.

Understanding how an amphicarpic species with a mixed mating system responds to fire: a population genetic approach.

Amphicarpic plants produce both above-ground and below-ground seeds. Because below-ground seeds are protected in the soil and may maintain viability when above-ground conditions are stressful, they were proposed as an adaptation to recolonize a site after disturbance. However, whether below-ground seeds are the main colonizers after a disturbance remains unknown. Our goal was to understand whether recolonization by an amphicarpic species after fire was accomplished primarily through germination of seeds produced above-ground or below-ground. We investigated Polygala lewtonii, an amphicarpic, perennial species endemic to fire-prone Florida sandhill and scrub, where fire kills plants but subsequently increases recruitment and population sizes. Polygala lewtonii produces three flower types: above-ground chasmogamous flowers and above-ground and below-ground cleistogamous flowers, with previous research demonstrating chasmogamous flowers produce a much greater proportion of seeds than cleistogamous flowers. We quantified outcrossing in seeds produced by chasmogamous flowers to determine whether it differed from the 100 % self-fertilized below-ground seeds. Approximately 25 % of seeds from chasmogamous flowers showed evidence of cross-pollination. Assuming that chasmogamous flowers produce the majority of the above-ground seeds, as was shown previously, this indicates it is possible to differentiate between germination by above-ground versus below-ground seeds in post-fire colonization. We next compared genetic diversity, admixture, inbreeding and population genetic structure pre- and post-fire. If fire promoted germination of chasmogamous seeds, heterozygosity and admixture would increase, and genetic structure and inbreeding would decrease. Instead, inbreeding and genetic structure increased and admixture decreased, suggesting that the below-ground selfed seeds (with limited dispersal ability) increased their contribution to the population after fire, possibly because fire reduced above-ground seed viability. Additionally, new alleles not found previously in range-wide analyses emerged from the seed bank post-fire. These results suggest that amphicarpy is a powerful adaptation to preserve genetic variation, maintain adaptive potential and promote rapid post-fire colonization.

Seasonal and Simultaneous Cleistogamy in Rostrate Violets (Viola, subsect. Rostratae, Violaceae).

The special mixed reproductive system, i.e., the ability of an individual plant to develop both open, chasmogamous (CH) flowers adapted to cross-pollination and closed, cleistogamous (CL) flowers with obligate self-pollinating, is a common phenomenon in Viola L. In most sections of Northern Hemisphere violets, cleistogamy is seasonal, and CH and CL flowers develop sequentially in the season. Non-seasonal cleistogamy (simultaneous) is a rare phenomenon in rostrate violets. In the current study, we focused on modification of the CH/CL mating system in V. caspia by environmental conditions, resulting in a gradual switch from temporal cleistogamy, occurring in nature, to simultaneous cleistogamy under greenhouse conditions. V. reichenbachiana with seasonal cleistogamy was the control for V. caspia with the labile seasonal/simultaneous cleistogamy system. In simultaneous cleistogamy, the CH and CL flowers, fruits and seeds developed on an individual plant at the same time on the same branch. The typical difference between CH and CL flowers’ pistils is a straight style ending with a head-like stigma in CH and a curved style in CL adapted to self-pollination. This trait persists in the fruit and seed stages, allowing for easy recognition of fruit of CL and CH flowers in simultaneous cleistogamy. Floral meristems of CH flowers of V. reichenbachiana developed on the rhizome at the end of the growing season under short-day conditions and remained dormant until the following season. The CL floral meristems formed under long-day conditions on elongating lateral branches in the upper leaf axils. The daily temperature influenced the variable CH/CL ratio of V. caspia in nature and greenhouse conditions. Regulation of the CL/CH flower ratio by modifying environmental factors is important for basic research on genetic/epigenetic regulation of cleistogamy and for practical use to produce genetically stable lines of economically important species via CL seeds.

DOES CLEISTOGAMOUS SELF-FERTILIZATION PROVIDE REPRODUCTIVE ASSURANCE AGAINST SEED PREDATION IN RUELLIA HUMILIS?

Mixed mating is thought to be adaptive in many species of flowering plants because self-fertilization provides reproductive assurance when pollinator densities are low. In species with dimorphic flowers, automatic self-fertilization by small cleistogamous (CL) flowers might also provide reproductive assurance by avoiding floral herbivory. We tested this hypothesis by quantifying flower production, seed production, and seed predation across an entire reproductive season within a Missouri population of Ruellia humilis, a species with mixed mating enforced by dimorphic flowers. The production of CL flowers was delayed relative to the production of open-pollinated chasmogamous (CH) flowers, consistent with the hypothesis of reproductive assurance. The rate of seed predation by larvae of the Noctuid moth Tripudia rectangula was much higher for fruits produced by CH flowers (69%) than by CL flowers (18%). Thus, although CH flowers self-pollinate in the absence of outcross pollen, CL flowers are a much more economical source of selfed seeds, not only because they require fewer resources but also because they experience much lower levels of seed predation. Seed predation by Tripudia is common in many other cleistogamous species of Ruellia native to the southern United States and Mexico and might have played a significant role in the evolution of CL flowers in this large genus.

Two new species of Octomeria (Pleurothallidinae) from Cordillera del Cóndor, Zamora Chichinpe, Ecuador

Two new species of orchids, Octomeria pacii, and O. panguiensis are described and illustrated from Cordillera del Cóndor, Ecuador; likewise, information is provided on their distribution, habitat, and conservation status. These species differ from other species in the genus by their floral and vegetative characteristics and their apparent reproductive strategy of cleistogamy. Octomeria pacii is similar to Octomeria deceptrix, but differs in the caespitose habit, the ramicauls compressed along their entire length, the narrowly elliptical leaves, the autogamous flowers, the translucent white sepals suffused with magenta and the obovate, white lip, marked with magenta. Octomeria panguiensisis similar to O. werneri, but differs by the larger, shortly repent plants with lanceolate leaves, the cleistogamous flowers, the translucent white-colored sepals, the 5-veined lateral sepals and the oblong-trilobed lip with the apical lobe tridentate
 Keywords: self-compatibility, self-pollination, cleistogamy, Octomeria deceptrix, Octomeria werneri

Evolution: Cleistogamy to the rescue of zygomorphic flowers

Cleistogamous flowers never fully bloom and are thought to have evolved as a means to promote self-fertilisation. A new study reveals that this curious feature arose more frequently in flowers with bilateral symmetry.

Distribution Characteristics, Population Structure and Dynamics of the Endangered Plant, <i>Viola websteri</i> Hemsl.

Plant species exhibit current characteristics as a result of interactions with environmental conditions. The plants of Viola sp. have selected chasmogamous flowers with vigorous vegetative propagation or development of cleistogamous flowers as an adaptation strategy. Viola websteri is distributed on the Korean peninsula and the eastern part of Jilin Province, China. The center and edge of the distribution are expected to exhibit different population-dynamics. It is necessary to investigate the cause of its current limited distribution even though V. websteri has a mixed-mating strategy. Firstly, We examined the vegetation environment of habitats and evaluated its characteristics. Growth characteristics were examined through plant phenology. We then evaluated the population structure, characteristics of chasmogamous flowers, and productivity of cleistogamous flowers. Moreover, we compared population sizes between 2014 and 2018. Most habitats were located in deciduous broadleaf mixed forests adjacent to valleys. V. websteri produced chasmogamous flowers with self-incompatibility in April-May and cleistogamous flowers in June-September. The cleistogamous flower production is a strategy ensuring seed production under uncertain environmental fluctuations; these were approximately twice as numerous as chasmogamous flowers. The population structure was distinguished into stable and very unstable regions. There were sites where the population experienced a sharp decline in the 2018 compared to that of 2014. This large decline was found in the edge populations. The habitats had different microsites depending on the natural disturbances of drought and the matrix constituting the habitat, thus supporting various plants. Ensuring the production of seeds through cleistogamous flowers, it was determined that rapid seedling re-establishment and population replenishment were possible when the natural disturbance factor was removed. Environmental factors did not equally affect all populations or individuals. Therefore, it was expected that it would be able to persisted in a long time, despite the rapid decrease in the number of individuals in the population regionally. Local extinction and re-establishment are likely to repeat according to environmental change. We propose the additional population investigation based on this works are required. We also suggest a need to assess the long-term population dynamics and the genetic characteristics of chasmogamous flowers and cleistogamous flowers to establish and implement effective conservation strategies.

Repeated evolution of a reproductive polyphenism in plants is strongly associated with bilateral flower symmetry

Polyphenisms are a special type of phenotypic plasticity in which the products of development are not continuous but instead are separate and distinct phenotypes produced in the same genetic background. One of the most widespread polyphenisms in the flowering plants is cleistogamy, in which the same individual plant produces both open, cross-pollinated flowers as well as highly reduced and closed, self-pollinated (cleistogamous) flowers.1-5 Cleistogamy is not a rare evolutionary phenomenon. It has evolved independently at least 41 times.1 But what favors the evolution of cleistogamy is still largely unknown.1 Darwin6 proposed a hypothesis that has never been properly tested. He observed that cleistogamy is more common in taxa with bilaterally symmetric (zygomorphic) flowers than in those with radially symmetric (actinomorphic) flowers. Moreover, Darwin suggested that cleistogamous flowers help to ensure pollination, which he postulated is less certain in zygomorphic taxa that rely on more specialized groups of pollinators. Here, we combined the largest datasets on floral symmetry and cleistogamy and used phylogenetic approaches to show that cleistogamy is indeed disproportionately associated with zygomorphic flowers and that zygomorphic species are more likely to evolve cleistogamy than actinomorphic species. We also show that zygomorphic species are less capable of autonomous open-flower self-pollination (lower autofertility), suggesting that selection of cleistogamy via reproductive assurance in zygomorphic taxa could help account for Darwin's observation. Our results provide support for the hypothesis that polyphenisms are favored when organisms encounter contrasting environments.

Precise Genome Editing in miRNA Target Site via Gene Targeting and Subsequent Single-Strand-Annealing-Mediated Excision of the Marker Gene in Plants.

Gene targeting (GT) enables precise genome modification—e.g., the introduction of base substitutions—using donor DNA as a template. Combined with clean excision of the selection marker used to select GT cells, GT is expected to become a standard, generally applicable, base editing system. Previously, we demonstrated marker excision via a piggyBac transposon from GT-modified loci in rice. However, piggyBac-mediated marker excision has the limitation that it recognizes only the sequence TTAA. Recently, we proposed a novel and universal precise genome editing system consisting of GT with subsequent single-strand annealing (SSA)-mediated marker excision, which has, in principle, no limitation of target sequences. In this study, we introduced base substitutions into the microRNA miR172 target site of the OsCly1 gene—an ortholog of the barley Cleistogamy1 gene involved in cleistogamous flowering. To ensure efficient SSA, the GT vector harbors 1.2-kb overlapped sequences at both ends of a selection marker. The frequency of positive–negative selection-mediated GT using the vector with overlapped sequences was comparable with that achieved using vectors for piggyBac-mediated marker excision without overlapped sequences, with the frequency of SSA-mediated marker excision calculated as ~40% in the T0 generation. This frequency is thought to be adequate to produce marker-free cells, although it is lower than that achieved with piggyBac-mediated marker excision, which approaches 100%. To date, introduction of precise substitutions in discontinuous multiple bases of a targeted gene using base editors and the prime editing system based on CRISPR/Cas9 has been quite difficult. Here, using GT and our SSA-mediated marker excision system, we succeeded in the precise base substitution not only of single bases but also of artificial discontinuous multiple bases in the miR172 target site of the OsCly1 gene. Precise base substitution of miRNA target sites in target genes using this precise genome editing system will be a powerful tool in the production of valuable crops with improved traits.

Insights into amphicarpy from the compact genome of the legume Amphicarpaea edgeworthii.

Amphicarpy (seed heteromorphy) is a unique and fascinating reproductive strategy wherein a single plant produces both aerial and subterranean fruits. This strategy is believed to be an adaptation to life under stressful or uncertain environments. Here, we sequenced and de novo assembled a chromosome‐level genome assembly of the legume Amphicarpaea edgeworthii Benth. The 299‐Mb A. edgeworthii genome encodes 27 899 protein‐coding genes and is the most compact sequenced legume genome reported until date. Its reduced genome size may be attributed to the reduced long‐terminal repeat retrotransposon content, which stems from the unequal homologous recombination. Gene families related to immunity and stress resistance have been contracted in A. edgeworthii, which is consistent with the notion that the amphicarpic reproductive strategy may be a complementary mechanism for its weak environmental‐adaptation ability. We demonstrated the ‘ABCE’ model for the differentiation of chasmogamous and cleistogamous flowers. In addition, the characteristics of aerial and subterranean seeds in hard‐seededness were explored. Thus, we suggest that the A. edgeworthii genome, which is the first of an amphicarpic plant, offers significant insights into its unusual reproductive strategy that is a key resource towards comprehending the evolution of angiosperms.

Gibberellins are required for dimorphic flower development in Viola philippica

Photoperiod is a major determinant of chasmogamous (CH)–cleistogamous (CL) dimorphic flower development in Viola philippica, and only long-day (LD) conditions induce CL flowers. In this study, it was found that the active gibberellin (GA) content in CL floral buds was higher than in CH floral buds formed under short-day (SD) conditions, suggesting that the biosynthesis of active GAs is enhanced by a longer photoperiod and may be associated with dimorphic flower development. Thus, the next step was to molecularly characterize the key V. philippica GA synthesis genes GA 20-oxidase (VpGA20ox) and GA 3-oxidase (VpGA3ox). In terms of the expression of VpGA20ox and VpGA3ox, it was found that the active GAs could be upregulated in developing pistils under both LD and SD conditions to develop functional pistils, and GAs could also accumulate in the stamens under SD conditions. The anthers and the adjacent petals were well developed under SD conditions. In contrast, the above-mentioned floral organs displayed low GA contents under LD conditions and were poorly developed. Although the application of paclobutrazol, an inhibitor of GA synthesis, did not reverse CL development under LD conditions, exogenous GAs could partially trigger the transition from CH to CL flowers under relative SD conditions (≤12 h daylight). This was coupled with the downregulation of B-class MADS-box genes, thereby restraining stamen and petal development. Both VpGA20ox and VpGA3ox exhibited similar expression profiles with B-class MADS-box genes in the development of the stamens and petals. Therefore, in response to photoperiod, GA signaling could affect the expression of B-class homeotic genes and regulate dimorphic flower development in Viola. As a compensation for poorly-developed nectaries, anthers, and petals, filament elongation, style shortness, and inward bending could ensure self-pollination in CL flowers. This work provides new insights into the regulation of CH–CL floral development and the evolutionary significance of the formation of dimorphic flowers.

Plantago australis (Plantaginaceae) produces both chasmogamous and cleistogamous flowers: Field work, herbarium and literature-based evidence

Plantago australis is considered to be obligately cleistogamous. Here we present evidence based on field work, herbarium collections and literature to show that this species produces both chasmogamous and cleistogamous flowers. We also hypothesise that the species is ambophilic.

The genome of Cleistogenes songorica provides a blueprint for functional dissection of dimorphic flower differentiation and drought adaptability.

Summary Cleistogenes songorica (2n = 4x = 40) is a desert grass with a unique dimorphic flowering mechanism and an ability to survive extreme drought. Little is known about the genetics underlying drought tolerance and its reproductive adaptability. Here, we sequenced and assembled a high‐quality chromosome‐level C. songorica genome (contig N50 = 21.28 Mb). Complete assemblies of all telomeres, and of ten chromosomes were derived. C. songorica underwent a recent tetraploidization (~19 million years ago) and four major chromosomal rearrangements. Expanded genes were significantly enriched in fatty acid elongation, phenylpropanoid biosynthesis, starch and sucrose metabolism, and circadian rhythm pathways. By comparative transcriptomic analysis we found that conserved drought tolerance related genes were expanded. Transcription of CsMYB genes was associated with differential development of chasmogamous and cleistogamous flowers, as well as drought tolerance. Furthermore, we found that regulation modules encompassing miRNA, transcription factors and target genes are involved in dimorphic flower development, validated by overexpression of CsAP2_9 and its targeted miR172 in rice. Our findings enable further understanding of the mechanisms of drought tolerance and flowering in C. songorica, and provide new insights into the adaptability of native grass species in evolution, along with potential resources for trait improvement in agronomically important species.

Translational Pigeonpea Genomics Consortium for Accelerating Genetic Gains in Pigeonpea (Cajanus cajan L.)

Pigeonpea is one of the important pulse crops grown in many states of India and plays a major role in sustainable food and nutritional security for the smallholder farmers. In order to overcome the productivity barrier the Translational Pigeonpea Genomics Consortium (TPGC) was established, representing research institutes from six different states (Andhra Pradesh, Karnataka, Madhya Pradesh, Maharashtra, Telangana, and Uttar Pradesh) of India. To enhance pigeonpea productivity and production the team has been engaged in deploying modern genomics approaches in breeding and popularizing modern varieties in farmers’ fields. For instance, new genetic stock has been developed for trait mapping and molecular breeding initiated for enhancing resistance to fusarium wilt and sterility mosaic disease in 11 mega varieties of pigeonpea. In parallel, genomic segments associated with cleistogamous flower, shriveled seed, pods per plant, seeds per pod, 100 seed weight, and seed protein content have been identified. Furthermore, 100 improved lines were evaluated for yield and desirable traits in multi-location trials in different states. Furthermore, a total of 303 farmers’ participatory varietal selection (FPVS) trials have been conducted in 129 villages from 15 districts of six states with 16 released varieties/hybrids. Additionally, one line (GRG 152 or Bheema) from multi-location trials has been identified by the All India Coordinated Research Project on Pigeonpea (AICRP-Pigeonpea) and released for cultivation by the Central Variety Release Committee (CVRC). In summary, the collaborative efforts of several research groups through TPGC is accelerating genetics gains in breeding plots and is expected to deliver them to pigeonpea farmers to enhance their income and improve livelihood.

Phylogenomics of the aquatic plant genus Ottelia (Hydrocharitaceae): Implications for historical biogeography

Ottelia Pers. is the second largest genus of the family Hydrocharitaceae, including approximately 23 extant species. The genus exhibits a diversity of both bisexual and unisexual flowers, and complex reproductive system comprising cross-pollinated to cleistogamous flowers. Ottelia has been regarded as a pivotal group to study the evolution of Hydrocharitaceae, but the phylogenic relationships and evolutionary history of the genus remain unresolved. Here, we reconstructed a robust phylogenetic framework for Ottelia using 40 newly assembled complete plastomes. Our results resolved Ottelia as a monophyletic genus consisting of two major clades, which correspond to the main two centers of diversity in Asia and Africa. According to the divergence time estimation analysis, the crown group Ottelia began to diversify around 13.09 Ma during the middle Miocene. The biogeographical analysis indicated the existence of the most recent common ancestor somewhere in Africa/Australasia/Asia. Basing on further insights from the morphological evolution of Ottelia, we hypothesized that the ancestral center of origin was in Africa, from where the range expanded by transoceanic dispersal to South America and Australasia, and further from Australasia to Asia. We suggested that the climatic change and global cooling since the mid-Miocene, such as the development of East Asian monsoon climate and tectonic movement of the Yunnan-Guizhou Plateau (YGP), might have played a crucial role in the evolution of Ottelia in China.

Biological peculiarities and cultivation of groundnut (a review)

Peanut is one of the most important crops in the Fabaceae Lindl. (Leguminosae L.) family. South America is considered to be the homeland of peanut, but now this crop is cultivated in America, Africa, Australia, Europe and Asia. The modern phylogenetic system of the genus Arachis L. includes 79 wild species and one cultivated species of common peanut (A. hypogaea L.). Diploid species contain 2n = 20 chromosomes of the A, B or D genome, tetraploids have A and B genomes. The А and В genomes are sequenced. Special biological features of all peanut varieties are the presence of chasmogamous and cleistogamous flowers and the development of pods only underground (geocarpy). Along with high requirements for improving the quality of oil and food products, much attention is paid to their safety: resistance to aflatoxin contamination and mitigation of allergenicity. Peanut cultivars vary in plant habit, shape and color of pods and seeds. Their growing season in Africa, Latin America and Asia is from 160 to 200 days, so early-ripening forms need to be selected for the south of the Russian Federation. Breeders from the Pustovoit Institute of Oil Crops (VNIIMK) have developed peanut cultivars with a yield of 2.0–3.3 t/ha and growing season duration of 115–120 days, adaptable to the environments of Krasnodar Territory. At present, there is no large-scale peanut production in Russia, nor any breeding efforts are underway. As for the world, along with conventional breeding practices (individual selection, intra- and interspecies crosses, etc.), peanut is widely involved in genomic studies. A number of cultivars highly resistant to pests, diseases and drought have been released. Over 15,000 peanut accessions are preserved in the world’s gene banks, including 1823 accessions in the collection of the Vavilov Institute (VIR). Utilization of the worldwide genetic resources of peanut and use of modern research technologies will contribute to the revival of peanut cultivation in Russia.

Comparative phylogeography, morphological boundaries and climate envelopes of two sympatric widespread Bromeliaceae from the southern Andes

AbstractWe studied the epiphytic species Tillandsia capillaris and T. virescens that grow as neotenic forms with autogamous and cleistogamous flowers. They have expanded in Andean mountain environments, where they grow sympatrically across most of their ranges in arid regions of Peru, central-western Bolivia and north-central Argentina and Chile. We studied the relationships between the two species and populations within each of them, the morphological boundaries and the climatic characterization of distinct genetic entities. We analysed 125 and 102 individuals from 25 and 27 populations of T. virescens and T. capillaris, respectively, by means of haplotype networks, phylogenetic and genetic structure analyses, using the plastid gene ycf1 (c. 4500 bp) to define genetic groups. Morphological studies by discriminant analyses and correlation with climatic variables extracted from WorldClim were used to test differentiations among the genetic groups within each species. We confirm that T. capillaris is a distinct entity and that T. virescens consists of three groups that diverged by vicariance from widespread ancestors. The high genetic diversity found in both species is consistent with the complex geography of the Andes and the Pleistocene glaciation cycles that have driven ecological speciation in both species reflected in the timing of divergence of the clusters.

Environmental impact on the temporal production of chasmogamous and cleistogamous flowers in the mixed breeding system of Viola pubescens.

Viola pubescens is a perennial, mixed breeding herb that produces both chasmogamous and cleistogamous flowers at different times of the season. Once bud type is specified, it does not convert from one form to the other. While temporal production of the two flowers is known to be influenced by environmental factors, the specific environmental cues that signal emergence of each flower type have not been empirically studied. To investigate the environmental parameters driving seasonal development of chasmogamous versus cleistogamous flowers, a native V. pubescens population was examined during the spring and summer of 2016 and 2017. Measurements of light quantity, canopy cover, photoperiod, temperature, soil moisture, soil pH, and the number of chasmogamous and cleistogamous buds were collected on either a weekly or biweekly basis. Independent zero-inflated negative binomial (ZINB) regressions were used to model the odds of bud production (0 versus 1 bud) and bud counts (≥ 1 bud) as a function of the environmental variables. Results of the ZINB models highlight key differences between the environmental variables that influence chasmogamous versus cleistogamous bud development and counts. In addition to the ZINB regressions, individual logistic regressions were fit to the bud data. The logistic models support results of the ZINB models and, more crucially, identify specific environmental thresholds at which each bud type is probable. Collectively, this work offers novel insight into how environmental variables shape temporal development of chasmogamous and cleistogamous flowers, suggests distinct threshold values that may aid in selectively inducing each flower type, and provides insight into how climatic change may impact mixed breeding species.

Analysis of Six Transcription Factor Families Explores Transcript Divergence of Cleistogamous and Chasmogamous Flowers in Cleistogenes songorica.

Cleistogenes songorica is a cultivated turfgrass species that employs a mixed breeding system. To determine the morphological differences and molecular mechanisms of the chasmogamous (CH) and cleistogamous (CL) flowers of this species, we evaluated seed traits and analyzed six transcription factor (TF) families related to floral development. The seed traits from the CH and CL flowers were significantly different. In total, 12 CsAP2, 13 CsSPL, 9 CsGRF, 21 CsMYB, 15 CsMADS, and 1 CsLFY differentially expressed genes were identified from the transcriptome of the C. songorica flowers, which was further supported by evolutionary relationships and conserved motifs. All collinear gene pairs had a Ka/Ks ratio <1. Analysis of the promoters and miRNAs of the TFs revealed that the members of the six TF families may coregulate the divergence of CH and CL flowers during evolution. Two CsAP2, 8 CsSPL, 6 CsGRF, 3 CsMYB (targeted by miR172s, miR156s, miR396a/b and miR159a/b, respectively), and 15 CsMADS genes as well as 1 CsLFY gene may be involved in the development of CH and CL flowers. This study is the first to analyze the differences between CH and CL flowers at the TF-family level, which will help in the understanding of dimorphic turfgrasses.

Season‐dependent effect of cleistogamy in Lamium amplexicaule: flower type origin versus inbreeding status

Inbreeding depression is traditionally considered a major factor favoring outcrossing in hermaphrodites. Recent experiments have shown that environmental conditions can influence the magnitude of inbreeding depression, but their relevance in natural populations is unclear. To investigate this, we studied the cleistogamous species Lamium amplexicaule, an annual species with individuals experiencing either spring or autumn environmental conditions. In this species, the proportion of cleistogamous/chasmogamous flowers changes according to seasonal cues (e.g., temperature, photoperiod). Our hypothesis was that the plasticity of cleistogamy is an adaptation to seasonal fitness variation in different flower progenies. To test this hypothesis, we produced the three possible types of progenies through controlled crosses: (1) selfed progeny from cleistogamous flowers; (2) chasmogamous selfed progeny; and (3) chasmogamous outcrossed progeny. Progeny fitness was then measured in a common garden in the two reproduction seasons (autumn and spring). The results showed that season had a major impact on fitness. The fitness of the different progeny types changed across seasons, indicating that the effect of cleistogamy on progeny performance is season-dependent, consistent with a previous study in a similar environment. Surprisingly, the flower from which the progeny issued (cleistogamous or chasmogamous) had more impact on fitness than the inbred status of the progeny (selfed versus outcrossed), suggesting a potential role of epigenetic processes. The observed patterns of variation were not consistent either with adaptation to environment-dependent inbreeding depression or to variation in resource availability, but were possibly consistent with adaptation to seasonal pollinator activity.