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Balancing selection is a form of natural selection maintaining diversity at the sites it targets and at linked nucleotide sites. Due to selection favoring heterozygosity, it has the potential to facilitate the accumulation of a "sheltered" load of tightly linked recessive deleterious mutations. However, precisely evaluating the extent of these effects has remained challenging. Taking advantage of plant self-incompatibility as one of the best-understood examples of long-term balancing selection, we provide a highly resolved picture of the genomic extent of balancing selection on the sheltered genetic load. We used targeted genome resequencing to reveal polymorphism of the genomic region flanking the self-incompatibility locus in three sample sets in each of the two closely related plant species Arabidopsis halleri and Arabidopsis lyrata , and used 100 control regions from throughout the genome to factor out differences in demographic histories and/or sample structure. Nucleotide polymorphism increased strongly around the S -locus in all sample sets, but only over a limited genomic region, as it became indistinguishable from the genomic background beyond the first 25–30 kb. Genes in this chromosomal interval exhibited no excess of mutations at 0-fold degenerated sites relative to putatively neutral sites, hence revealing no detectable weakening of the efficacy of purifying selection even for these most tightly linked genes. Overall, our results are consistent with the predictions of a narrow genomic influence of linkage to the S -locus and clarify how natural selection in one genomic region affects the evolution of the adjacent genomic regions. [ABSTRACT FROM AUTHOR]

Self-incompatibility (SI) is a genetic mechanism in many flowering plants by which generative reproduction is prevented. The self-incompatibility caused by the genetic functions of the cell is controlled by genes called S genes or self-incompatibility genes. Self-incompatibility results in decreased pollination and ultimately yield loss. In apple (Malus domestica L.), self-incompatibility is controlled by multi-allelic S-locus. Approaches in the S-glycoprotein profiles and allele-specific PCR methods using the gene profiles and S-glycoprotein profiles for determination of the incompatibility levels are of great importance. In current study, the self-incompatibility status of 192 apple genotypes (such as, Amasya, Hüryemez, Şah elması, Tokat, Demir elması etc.) obtained from the National Collection of Atatürk Horticultural Central Research Institute, Yalova, Turkey, has been determined. For this purpose, genotype-specific allele status and compatibility levels were screened via PCR (Polymerase Chain Reaction) using 4 different S-alleles (Sd, Sf, S26 and S9). 181 genotypes containing at least 1 S-allele were identified as ‘Partially Incompatible’ and 12 genotypes involving 4 S-alleles were assigned ‘Totally Incompatible’. No S-alleles were observed in 2 genotypes (Pancarlık and Hüryemez) which exhibited ‘Compatibility’ status.

Plants employ self-incompatibility (SI) to promote cross-fertilization. In Brassicaceae, this process is regulated by the formation of a complex between the pistil determinant S receptor kinase (SRK) and the pollen determinant S-locus protein 11 (SP11, also known as S-locus cysteine-rich protein, SCR). In our previous study, we used the crystal structures of two eSRK–SP11 complexes in Brassica rapa S8 and S9 haplotypes and nine computationally predicted complex models to demonstrate that only the SRK ectodomain (eSRK) and SP11 pairs derived from the same S haplotype exhibit high binding free energy. However, predicting the eSRK–SP11 complex structures for the other 100 + S haplotypes and genera remains difficult because of SP11 polymorphism in sequence and structure. Although protein structure prediction using AlphaFold2 exhibits considerably high accuracy for most protein monomers and complexes, 46% of the predicted SP11 structures that we tested showed

In plants, macroautophagy/autophagy is an essential mechanism responsible for a large variety of processes throughout the plant’s lifecycle, including nutrient processing, immunity, stress responses and senescence. Previous studies had observed the presence of autophagosomes in an Arabidopsis sexual reproduction system that prevents self-fertilization (self-incompatibility), but their requirement in this pathway was unclear. Using autophagy-deficient mutants, we have recently found that autophagy is a key contributor in the Arabidopsis self-incompatibility response to reject self-pollen.

Results of a study of self-pollination in two of Ukrainian flora's threatened species, Gladiolus imbricatus and Iris sibirica (Iridaceae), are presented. To confirm or refute the occurrence of self-pollination in G. imbricatus and I. sibirica, an experiment was conducted on pollen germination on their stigmas. It has been found that in G. imbricatus pollination with autogenous pollen grains on the last day of flowering leads to the growth of pollen tubes in the cases of both hand and natural pollination. However, pollen grains on the stigma of an isolated I. sibirica flower were not detected on the first or last day of flowering, which means the absence of a mechanism of autonomous pollen transfer within the meranthium. Ungerminated pollen grains were also found on the stigma after autogenous hand pollination. It has been experimentally confirmed that in the absence of pollinators at the end of the flowering phase in G. imbricatus, as well as in some other members of the genus, self-pollination and self-fertilization are possible, which contribute to the wellbeing of populations and species. With regard to I. sibirica, it has been found that the autogamous self-incompatibility inherent in this species prevents inbreeding, maintaining heterozygosity in plant populations, allowing plants' better adaptation to different environmental conditions.

Camellia oleifera is a woody edible oil species with late self-incompatibility characteristics. Previous transcriptome analysis showed that genes involved in jasmonic acid signal transduction were significantly different in self-and cross-pollinated pistils of Camellia oleifera. To investigate the relationship between jasmonate signal and self-incompatibility by studying the core genes of jasmonate signal transduction. The results showed that exogenous JA and MeJA at 1.0 mM significantly inhibited pollen tube germination and pollen tube elongation. and JA up-regulated CoCOI1, CoJAZ1, and CoMYC, the core genes of jasmonate signal transduction. Subcellular localization indicated that CoCOI1 and CoJAZ1 were located in the nucleus and CoMYC2 in the endoplasmic reticulum. The three genes exhibited tissue-specific expression pattern. CoCOI1 was significantly expressed in pollen, CoJAZ1 was significantly expressed in ovary, CoMYC2 was significantly expressed in filaments, but not in pollen. Furthermore, CoJAZ1 and CoMYC2 were highly expressing at 24 h in self-pollinated styles. These results suggested that JA signal transduction of C. oleifera was involved in the process of self-pollination, and thus in the process of plant defense. When pollen tubes grew slowly in the style, ovary may receive JA signal, which initiates the molecular mechanism of inhibiting the growth of self-pollinating pollen tubes.

Simple polyembryony, where one gametophyte produces multiple embryos with different sires but the same maternal haplotype, is common among vascular plants. We develop an infinite-sites, forward population genetics model showing that together polyembryony's two benefits—"reproductive compensation" achieved by providing a backup for inviable embryos and the opportunity to favor the fitter of surviving embryos—can favor its evolution. Our model tests how these factors can favor the evolution of polyembryony and how these underlying benefits of polyembryony shape the genetic load under a range of biological parameters. While these two benefits are difficult to disentangle in nature, we construct variant models of polyembryony that either only include or only exclude the opportunity for reproductive compensation. We find that reproductive compensation strongly favors the evolution of polyembryony and that polyembryony is favored much more weekly in its absence, suggesting that the benefit of a backup embryo is the major force favoring polyembryony. Remarkably, we find nearly identical results in cases in which mutations impact either embryonic or postembryonic fitness (no pleiotropy) and in cases in which mutations have identical effects on embryonic and postembryonic fitness (extreme pleiotropy). Finally, we find that the consequences of polyembryony depend on its function—polyembryony results in a decrease in mean embryonic fitness when acting as a mechanism of embryo compensation and ultimately increases mean embryonic fitness when we exclude this potential benefit. [ABSTRACT FROM AUTHOR]

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The sweet cherry plant (Prunus avium L.) is primarily self-incompatible, with so-called S-alleles responsible for the inability of flowers to be pollinated not only by their own pollen grains but also by pollen from other cherries having the same S-alleles. This characteristic has wide-ranging impacts on commercial growing, harvesting, and breeding. However, mutations in S-alleles as well as changes in the expression of M locus-encoded glutathione-S-transferase (MGST) can lead to complete or partial self-compatibility, simplifying orchard management and reducing possible crop losses. Knowledge of S-alleles is important for growers and breeders, but current determination methods are challenging, requiring several PCR runs. Here we present a system for the identification of multiple S-alleles and MGST promoter variants in one-tube PCR, with subsequent fragment analysis on a capillary genetic analyzer. The assay was shown to unequivocally determine three MGST alleles, 14 self-incompatible S-alleles, and all three known self-compatible S-alleles (S3′, S4′, S5′) in 55 combinations tested, and thus it is especially suitable for routine S-allele diagnostics and molecular marker-assisted breeding for self-compatible sweet cherries. In addition, we identified a previously unknown S-allele in the 'Techlovicka´ genotype (S54) and a new variant of the MGST promoter with an 8-bp deletion in the ´Kronio´ cultivar. [ABSTRACT FROM AUTHOR]

Most seed plants produce both pollen and ovules. In principle, pollen export could interfere with pollen import through self‐pollination, resulting in ovule usurpation and reduced fruit set. Evidence for such interference exists under experimental settings but its importance under natural conditions is unknown. To test for sexual interference in nature, it is necessary to study together mating system, through paternity analyses, and fruit set, the proportion of flowers giving seeds or fruits. We developed a new model combining both processes, using chestnut (Castanea) as case study. We carried out a paternity analysis in an intensively studied plot of 273 trees belonging to three interfertile chestnut species and including a range of individuals with more or less functional stamens, resulting in a large data set of 1924 mating events. We then measured fruit set on 216 of these trees. Fruit set of male‐fertile trees was much lower than that of male‐sterile trees. Our process‐based model shows that pollen is not limiting in the study site and hence cannot account for reduced fruit set. It also indicates that self‐pollination is high (74%) but selfing rate is low (4%). Self‐pollen is less competitive than cross‐pollen, reducing sexual interference, but not sufficiently, as many ovules end up being self‐fertilized, 95% of which abort before fruit formation, resulting in the loss of 46% of the fruit crop. These results suggest that the main cause of reduced reproductive potential in chestnut is sexual interference by self‐pollen, raising questions on its evolutionary origins. [ABSTRACT FROM AUTHOR]

Self-incompatibility systems based on self-recognition evolved in hermaphroditic plants to maintain genetic variation of offspring and mitigate inbreeding depression. Despite these benefits in diploid plants, for polyploids who often face a scarcity of mating partners, self-incompatibility can thwart reproduction. In contrast, self-compatibility provides an immediate advantage: a route to reproductive viability. Thus, diploid selfing lineages may facilitate the formation of new allopolyploid species. Here, we describe the mechanism of establishment of at least four allopolyploid species in Brassicaceae (Arabidopsis suecica, Arabidopsis kamchatica, Capsella bursa-pastoris, and Brassica napus), in a manner dependent on the prior loss of the self-incompatibility mechanism in one of the ancestors. In each case, the degraded S-locus from one parental lineage was dominant over the functional S-locus of the outcrossing parental lineage. Such dominant loss-of-function mutations promote an immediate transition to selfing in allopolyploids and may facilitate their establishment. [ABSTRACT FROM AUTHOR]

Self-incompatibility is the system that inhibits pollen germination and pollen tube growth by self-pollen. This trait is important for the breeding of Brassica and Raphanus species. In these species, self-incompatibility is governed by the S locus, which contains three linked genes (a set called the S haplotype), i.e., S-locus receptor kinase, S-locus cysteine-rich protein/S-locus protein 11, and S-locus glycoprotein. A large number of S haplotypes have been identified in Brassica oleracea, B. rapa, and Raphanus sativus to date, and the nucleotide sequences of their many alleles have also been registered. In this state, it is important to avoid confusion between S haplotypes, i.e., an identical S haplotype with different names and a different S haplotype with an identical S haplotype number. To mitigate this issue, we herein constructed a list of S haplotypes that are easily accessible to the latest nucleotide sequences of S-haplotype genes, together with revisions to and an update of S haplotype information. Furthermore, the histories of the S-haplotype collection in the three species are reviewed, the importance of the collection of S haplotypes as a genetic resource is discussed, and the management of information on S haplotypes is proposed. [ABSTRACT FROM AUTHOR]

The fitness of self-progeny individuals is inferior to that of their outcrossed counterparts, resulting in a reduction in a plant population’s ability to survive and reproduce. To prevent self‐fertilization, angiosperms with hermaphrodite flowers may exploit a variety of mechanisms, including synchronous dichogamy and self-incompatibility. Synchronous dichogamy involves two flowering morphs, with strict within-morph synchronization, thereby preventing not only autogamy and geitonogamy but also intra-morph mating. Self-fertilization is also prevented by self-incompatibility, a genetic mechanism that allows the identification and rejection of “self” pollen, thereby preventing both autogamy and geitonogamy. Here, I seek to provide a perspective of flowering in Ziziphus species exhibiting both synchronous (i.e., “Early” morph flowers open in the morning and “Late” morph flowers open in the afternoon) protandrous dichogamy (i.e., pollen dispersal before the stigma becomes receptive) and self-incompatibility.

Hybridisation between individuals of different species can lead to maladapted or inviable progeny due to genetic incompatibilities between diverging species. On the other hand, mating with close relatives, or self-fertilisation may lead to inbreeding depression. Thus, both too much or too little divergence may lead to problems and the organisms have to carefully choose mating partners to avoid both of these pitfalls. In plants this choice occurs at many stages during reproduction, but pollen-pistil interactions play a particularly important role in avoiding inbreeding and hybridisation with other species. Interestingly, the mechanisms involved in avoidance of selfing and interspecific hybridisation may work via shared molecular pathways, as self-incompatible species tend to be more ‘choosy’ with heterospecific pollen compared to self-compatible ones. This review discusses various prezygotic post-pollination barriers to interspecific hybridisation, with a focus on the mechanisms of pollen-pistil interactions and their role in the maintenance of species integrity.

The recognition of pollen and pistil in the self-incompatibility process is generally determined by the interaction between the pollen S gene and pistil S gene located at the S locus. However, the regulatory mechanism of self-incompatibility in goji remains unknown. In this study, we used the self-compatible strain ‘13–19’ and self-incompatible strain ‘xin9’ from Ningxia as parents to create an F1 hybrid population. Reciprocal cross-pollination was performed within the same plant to evaluate the self-compatibility of the parents and F1 progeny. The parents and progeny were subjected to whole-genome resequencing, and mixed pools of DNA were constructed using 30 self-compatible and 30 self-incompatible individuals. Association analysis using the SNP-index method and Euclidean distance was employed to identify the key candidate region of the S locus. The candidate region was further annotated using the Swiss-Prot database to identify genes within the region. Additionally, transcriptome sequencing data from different organs/tissues, as well as from pistils of self-compatible and self-incompatible strains at control (0 h), short (0.5 h), medium (8 h), and long (48 h) time points after self-pollination and cross-pollination, were analyzed to assess differential gene expression and screen for self-compatibility-related loci. Specific primers were designed for PCR amplification to determine the S-RNase genotypes of the extreme parents. The results revealed that the S locus in goji is located within a 32.2 Mb region on chromosome 2 that contains a total of 108 annotated genes. Differential expression analysis showed that ten genes, including Lba02g01064, were specifically expressed in stamens, with four of them annotated as F-box genes, potentially serving as determinants of self-compatibility in stamens. Lba02g01102 was exclusively expressed in pistils and annotated as an S-RNase gene, likely involved in self-compatibility. The expression of Lba02g01102 in pistils decreased after self-pollination and cross-pollination. Six candidate genes exhibited significant changes after self-pollination and cross-pollination. Both parents and progeny carried two S-RNase alleles, and the S-RNase genotypes showed a significant correlation with self-compatibility, with the self-compatible progeny containing the S8-RNase allele. The identification of the S locus in goji provides molecular markers for future marker-assisted breeding and offers genetic resources for studying the mechanism of self-incompatibility in goji, thus contributing to the improvement of goji varieties.

Pollen pistil interaction studies were carried out in six sunnhemp genotypes. Pollen viability percentage of the six genotypes ranged from 0.0 to 93.5% and pollen germination ranged from 0.0 to 84.0% at four different floral stages starting from bud to opening of a flower. The viable pollen grains were present at the later three stages of flower for globose type anthers, while the pollen grains were viable at the later two floral stages for heart shaped anthers. Stigma receptivity was high to very high during S3 and S4 stages. Artificial self-pollination and sib pollination with a mixture of pollen grains were carried out and in vivo pollen tube growth in the style was investigated using fluorescence microscopy. In self-pollinated pistils, eight hours after pollination, callose wall development was seen around the nucellus cell. However, under sib pollinated pistil, eight hours after pollination, pollen tube penetrated into embryo sac of an ovule. The formation of callose in self-fertilized pistil prevented fertilization and seed set. But sib pollinated flowers were fertilized normally and a seed set could be observed. The investigation proved the existence of selfincompatibility.

The evolution from outcrossing to selfing occurred recently across the eukaryote tree of life in plants, animals, fungi, and algae. Despite short-term advantages, selfing is hypothetically an evolutionary dead-end reproductive strategy. The tippy distribution on phylogenies suggests that most selfing species are of recent origin. However, dating such transitions is challenging yet central for testing this hypothesis. We build on previous theories to disentangle the differential effect of past changes in selfing rate or from that of population size on recombination probability along the genome. This allowed us to develop two methods using full-genome polymorphisms to (1) test if a transition from outcrossing to selfing occurred and (2) infer its age. The teSMC and tsABC methods use a transition matrix summarizing the distribution of times to the most recent common ancestor along the genome to estimate changes in the ratio of population recombination and mutation rates overtime. First, we demonstrate that our methods distinguish between past changes in selfing rate and demographic history. Second, we assess the accuracy of our methods to infer transitions to selfing approximately up to 2.5Ne generations ago. Third, we demonstrate that our estimates are robust to the presence of purifying selection. Finally, as a proof of principle, we apply both methods to three Arabidopsis thaliana populations, revealing a transition to selfing approximately 600,000 years ago. Our methods pave the way for studying recent transitions to self-fertilization and better accounting for variation in mating systems in demographic inferences.

The Gametophytic Self-Incompatibility (GSI) system in diploid potato (Solanum tuberosum L.) poses a substantial barrier in diploid potato breeding by hindering the generation of inbred lines. One solution is gene editing to generate self-compatible diploid potatoes which will allow for the generation of elite inbred lines with fixed favorable alleles and heterotic potential. The S-RNase and HT genes have been shown previously to contribute to GSI in the Solanaceae family and self-compatible S. tuberosum lines have been generated by knocking out S-RNase gene with CRISPR-Cas9 gene editing. This study employed CRISPR-Cas9 to knockout HT-B either individually or in concert with S-RNase in the diploid self-incompatible S. tuberosum clone DRH-195. Using mature seed formation from self-pollinated fruit as the defining characteristic of self-compatibility, HT-B-only knockouts produced little or no seed. In contrast, double knockout lines of HT-B and S-RNase displayed levels of seed production that were up to three times higher than observed in the S-RNase-only knockout, indicating a synergistic effect between HT-B and S-RNase in self-compatibility in diploid potato. This contrasts with compatible cross-pollinations, where S-RNase and HT-B did not have a significant effect on seed set. Contradictory to the traditional GSI model, self-incompatible lines displayed pollen tube growth reaching the ovary, yet ovules failed to develop into seeds indicating a potential late-acting self-incompatibility in DRH-195. Germplasm generated from this study will serve as a valuable resource for diploid potato breeding.

Self-incompatibility in Prunus species is governed by a single locus consisting of two highly multi-allelic and tightly linked genes, one coding for an F-box protein—i.e., SFB in Prunus- controlling the pollen specificity and one coding for an S-RNase gene controlling the pistil specificity. Genotyping the allelic combination in a fruit tree species is an essential procedure both for cross-based breeding and for establishing pollination requirements. Gel-based PCR techniques using primer pairs designed from conserved regions and spanning polymorphic intronic regions are traditionally used for this task. However, with the great advance of massive sequencing techniques and the lowering of sequencing costs, new genotyping-by-sequencing procedures are emerging. The alignment of resequenced individuals to reference genomes, commonly used for polymorphism detection, yields little or no coverage in the S-locus region due to high polymorphism between different alleles within the same species, and cannot be used for this purpose. Using the available sequences of Japanese plum S-loci concatenated in a rosary-like structure as synthetic reference sequence, we describe a procedure to accurately genotype resequenced individuals that allowed the analysis of the S-genotype in 88 Japanese plum cultivars, 74 of them are reported for the first time. In addition to unraveling two new S-alleles from published reference genomes, we identified at least two S-alleles in 74 cultivars. According to their S-allele composition, they were assigned to 22 incompatibility groups, including nine new incompatibility groups reported here for the first time (XXVII-XXXV). [ABSTRACT FROM AUTHOR]

SUMMARY: Gametophytic self‐incompatibility (GSI) has been widely studied in flowering plants, but studies of the mechanisms underlying pollen tube growth arrest by self S‐RNase in GSI species are limited. In the present study, two leucine‐rich repeat extensin genes in pear (Pyrus bretschneideri), PbLRXA2.1 and PbLRXA2.2, were identified based on transcriptome and quantitative real‐time PCR analyses. The expression levels of these two LRX genes were significantly higher in the pollen grains and pollen tubes of the self‐compatible cultivar 'Jinzhui' (harboring a spontaneous bud mutation) than in those of the self‐incompatible cultivar 'Yali'. Both PbLRXA2.1 and PbLRXA2.2 stimulated pollen tube growth and attenuated the inhibitory effects of self S‐RNase on pollen tube growth by stabilizing the actin cytoskeleton and enhancing cell wall integrity. These results indicate that abnormal expression of PbLRXA2.1 and PbLRXA2.2 is involved in the loss of self‐incompatibility in 'Jinzhui'. The PbLRXA2.1 and PbLRXA2.2 promoters were directly bound by the ABRE‐binding factor PbABF.D.2. Knockdown of PbABF.D.2 decreased PbLRXA2.1 and PbLRXA2.2 expression and inhibited pollen tube growth. Notably, the expression of PbLRXA2.1, PbLRXA2.2, and PbABF.D.2 was repressed by self S‐RNase, suggesting that self S‐RNase can arrest pollen tube growth by restricting the PbABF.D.2–PbLRXA2.1/PbLRXA2.2 signal cascade. These results provide novel insight into pollen tube growth arrest by self S‐RNase. Significance Statement: PbLRXA2.1 and PbLRX.A2.2 mediate pollen tube growth and attenuate the inhibitory effects of self S‐RNase on pollen tube growth by stabilizing the actin cytoskeleton and enhancing cell wall integrity. Self S‐RNase can arrest pollen tube growth by restricting the PbABF.D.2–PbLRXA2.1/PbLRXA2.2 signal cascade. [ABSTRACT FROM AUTHOR]

Premise: Seed production is frequently limited by the receipt of insufficient or low‐quality pollen, collectively termed "pollen limitation" (PL). In taxa with gametophytic self‐incompatibility (GSI), incompatible pollen can germinate on stigmas but pollen tubes are arrested in styles. This allows for estimates of pollen performance before, during, and after self‐recognition, as well as insight into the factors underlying pollen quality limitation in GSI taxa. Methods: We scored pollen performance following self and outcross pollinations in Argentina anserina to identify the location of self‐recognition and establish the relationship between pollen tubes and seed production. We then estimated quantity and quality components of PL from >3300 field‐collected styles. We combined our results with other studies to test the prediction that low pollen quality, but not quantity, drives higher PL in self‐incompatible (SI) taxa than in self‐compatible taxa (SC). Results: Self and outcross pollen germinated readily on stigmas, but 96% of germinated self‐pollen was arrested during early tube elongation. Reproduction in the field was more limited by pollen quality than by quantity, and pollen failure near the location of self‐recognition was a stronger barrier to fertilization than pollen germination. Across 26 taxa, SI species experienced stronger pollen quality, but not quantity, limitation than SC species. Conclusions: Evaluating pollen performance at multiple points within pistils can elucidate potential causes of pollen quality limitation. The receipt of incompatible pollen inhibits fertilization success more than insufficient pollen receipt or poor pollen germination in A. anserina. Likewise, pollen quality limitation drives high overall PL in other SI taxa. [ABSTRACT FROM AUTHOR]

Brassica vegetables are very important to human beings. Self-incompatibility (SI) is a common phenomenon in Brassica. Breeding by SI lines is an important way to utilize heterosis of Brassica vegetables. It is believed that the SI inheritance in Brassica species is controlled by three linkage genes on the S-locus, including SRK (S-locus receptor kinase), SCR (S-locus cystine-rich protein)/SP11 (S-locus protein 11), and SLG (S-locus glycoprotein). SRK is the female determinant and SCR/SP11 is the pollen S gene. The expression of SLG is necessary for SRK, and it enhances the SRK-mediated SI reaction. In addition to these three S-locus genes, some other functional molecules also have significant regulatory effects on SI, such as ARC1 (arm repeat containing 1), MLPK (M-locus protein kinase), Exo70A1 (exocyst compounds), THLl/THL2 (thioredoxin H-like), MOD (aquaporin), SLR (S-locus-related glycoprotein), BPCI (pollen calcium-binding protein I), etc. SI is also associated with the dominant/recessive relationship between S alleles. Here, the genetic elements and molecular mechanisms of SI, mainly in Brassica vegetables, are reviewed. [ABSTRACT FROM AUTHOR]

Reactive oxygen species (ROS), which are generated by different cellular compartments through aerobic metabolic pathways, are generally considered to play dual roles in plants. They are toxic compounds at high concentration but are also required to modulate multiple signaling reactions and gene regulatory networks in biotic and abiotic stress responses as well as during plant growth and development. Recent studies revealed that ROS play also critical roles in various aspects of flowering plant reproduction. In male gametophytes, ROS regulate germline specification and differentiation, and they are associated with programmed cell death (PCD) of the tapetum to regulate pollen development. During female gametophyte formation, ROS are involved in cell identity determination, polarity regulation and megaspore PCD. After pollination, ROS are of vital importance for regulation of self-incompatibility responses, for pollen hydration and for successful germination on the stigma. Further roles have been reported during pollen tube growth and guidance in the transmitting tract, pollen tube reception and sperm cell release in the female gametophyte as well as during gamete fusion. In this chapter, we will provide an overview of the multiple functions of ROS signaling during plant reproductive development from germline initiation to double fertilization. We will focus on flowering plants and discuss the generation and elimination of ROS, and the induction and regulation of various downstream signaling pathways. [ABSTRACT FROM AUTHOR]

Self-incompatibility (SI) is a genetic mechanism in many flowering plants by which generative reproduction is prevented. The self-incompatibility caused by the genetic functions of the cell is controlled by genes called S genes or self-incompatibility genes. Self-incompatibility results in decreased pollination and ultimately yield loss. In apple (Malus domestica L.), self-incompatibility is controlled by multi-allelic S-locus. Approaches in the S-glycoprotein profiles and allele-specific PCR methods using the gene profiles and S-glycoprotein profiles for determination of the incompatibility levels are of great importance. In current study, the self-incompatibility status of 192 apple genotypes (such as, Amasya, Hüryemez, Şah elması, Tokat, Demir elması etc.) obtained from the National Collection of Atatürk Horticultural Central Research Institute, Yalova, Turkey, has been determined. For this purpose, genotype-specific allele status and compatibility levels were screened via PCR (Polymerase Chain Reaction) using 4 different S-alleles (Sd, Sf, S26 and S9). 181 genotypes containing at least 1 S-allele were identified as 'Partially Incompatible' and 12 genotypes involving 4 S-alleles were assigned 'Totally Incompatible'. No S-alleles were observed in 2 genotypes (Pancarlık and Hüryemez) which exhibited 'Compatibility' status. [ABSTRACT FROM AUTHOR]

Extending the range of Osmanthus species into more cold-hardy climates would open new opportunities for adoption and use of these species by growers, landscapers, and the public. Breeding improvement is hindered by few available cultivars and a lack of female or perfect flowers. The objectives of this study were to evaluate floral morphology and pollination biology of Osmanthus species available in the U.S. market. Thirty-three genotypes representing four species were evaluated in McMinnville, TN, USA for genome size, floral morphology, pollen viability, and self- and cross-compatibility. All genotypes were diploid with 2C genome sizes ranging from 2.89 to 3.21 pg. Perfect flowers were observed in all O. armatus and 82% of O. heterophyllus genotypes. All observed O. fortunei and O. fragrans genotypes had male-only flowers. Pollen viability based on staining and in vitro germination ranged from 8% to 98% and 6% to 53%, respectively. Pollen germination was observed on stigmas of 94.2% of O. armatus and O. heterophyllus flowers collected 24 hours after cross- or self-pollination. There was a significant association between cross type and percentage of flowers with pollen tubes reaching the ovaries; after 72 h, pollen tubes had reached the ovaries of in 67% of intraspecific crosses, 78% of interspecific crosses, and 0% of self crosses (χ2 = 26.5, p < 0.001). This study provides evidence of a self-incompatibility system in O. armatus and O. heterophyllus and provides insights into opportunities and challenges for Osmanthus hybrid breeding. [ABSTRACT FROM AUTHOR]

Summary: Self‐incompatibility (SI) involves specific interactions during pollination to reject incompatible ('self') pollen, preventing inbreeding in angiosperms. A key event observed in pollen undergoing the Papaver rhoeas SI response is the formation of punctate F‐actin foci.Pollen tube growth is heavily energy‐dependent, yet ATP levels in pollen tubes have not been directly measured during SI. Here we used transgenic Arabidopsis lines expressing the Papaver pollen S‐determinant to investigate a possible link between ATP levels, cytosolic pH ([pH]cyt) and alterations to the actin cytoskeleton.We identify for the first time that SI triggers a rapid and significant ATP depletion in pollen tubes. Artificial depletion of ATP triggered cytosolic acidification and formation of actin aggregates. We also identify in vivo, evidence for a threshold [pH]cyt of 5.8 for actin foci formation. Imaging revealed that SI stimulates acidic cytosolic patches adjacent to the plasma membrane.In conclusion, this study provides evidence that ATP depletion plays a pivotal role in SI upstream of programmed cell death and reveals a link between the cellular energy status, cytosolic acidification and alterations to the actin cytoskeleton in regulating Papaver SI in pollen tubes. See also the Commentary on this article by Cheung, 236: 1625–1628. [ABSTRACT FROM AUTHOR]

Opuntia is the most diverse and widespread genus in the Cactaceae; several species have broad distribution ranges and have successfully adapted to distinct habitats. One explanation for their success is the diversity of their reproductive systems. Opuntia macrocentra is one of the most common widespread species in North American deserts. A previous work on a northern population documented that this species has a mixed‐mating system and receives a low frequency of pollinators. In a southern population of O. macrocentra, we described its floral visitors, anthesis schedules and flower morphometry. Through controlled pollination experiments (selfing, outcrossing, supplementary pollen and natural pollination), we assessed the breeding and mating systems and measured pollen limitation. Eight different bee species visited the flowers. Based on their conduct, Diadasia rinconis and Lasioglossum sp. were the main pollinators. Flowers were homogamous and herkogamous. Floral traits and pollination experiments showed that O. macrocentra is xenogamous and self‐incompatible. Fruit set of the natural pollination, outcrossing and supplementary pollen treatments were not statistically different. Seed set was higher in natural pollination and the pollen limitation index for seed set was negative. Based on these results, we conclude that O. macrocentra is not pollen limited in the studied population. These results contrast with those of the previously studied population; therefore, we caution about making generalizations about the reproductive system based solely on studies in one population. The diversity of mating systems displayed by O. macrocentra may explain its broad distribution and adaptation to different environmental factors, one of which may be pollinator availability. Our results highlight the need to describe the reproductive ecology of populations instead of species, because reproductive systems and ecological factors such as plant–animal interactions may differ across their distribution range. [ABSTRACT FROM AUTHOR]

In the production of blueberries, pollination is a key factor determining high quality yield. Problems with the process of pollination in blueberries occurs frequently. Lack of the effective pollination results in a lower yield and production of small fruits with low commercial value. The experiment described in this paper was carried out to test the effect of titanium organic complex (TOC) on the pollination and fertilization processes, seed formation, fruit setting and fruit development of blueberry in two consecutive years. The experiment consisted of two experimental factors: pollination type (self or open pollination) and application of TOC or lack of it. The presented study proves that TOC stimulates pollen adhesion and germination on the stigma, fruit setting, seed setting and fruit development. In self-pollinated flowers, the use of TOC increased the mass of each berry by 28% (in the first year) and 26% (in the second year), compared to non-treated plants. In open pollinated flowers, the application of TOC increased fruit mass by 38% (in the first year) and 29% (in the second year), compared to non-treated plants. Therefore, TOC may be treated as a substance stimulating the total fruit yield and mass of individual fruit. [ABSTRACT FROM AUTHOR]

ABSTRACT Distyly is a floral polymorphism that promotes cross-pollination through precise pollination. Psychotria is a mostly tropical genus of distylous Rubiaceae. Although widely studied in Brazil, some regions/species are still poorly explored, which hinders the understanding of distyly system along a greater geographical range. Here, we studied a subtropical population of Psychotria brachyceras in southern Brazil. For this, we characterized morphs occurrence, reciprocity of sexual structures, mating and incompatibility systems. In addition, we compared the reciprocity values from other species of the genus based on values gathered from literature to understand how the population behaves. The population showed equal proportions of thrum (short stigma/high anthers) and pin (high stigma/short anthers) morphs. Reciprocity was higher than the average for the genus and inaccuracy values between sexual organs were evenly distributed between the organ types, indicating a trend to typical distyly. Higher fruit set rates in intermorph and open pollination treatments and pollen tube growth observations confirmed that the population has a functional heteromorphic incompatibility system. Psychotria brachyceras showed no signs of distinct traits to typical distylous populations, which seems to ensure reproductive success and distyly maintenance.

The Japanese plum (Prunus salicina Lindl.) is a fruit tree globally cultivated in temperate regions of the world. Its floral biology and yield are affected by several factors, with issues related to self- and cross- (in) compatibility among varieties being emblematic of the whole Rosaceae family. The aim of this work was to elucidate the fruit set, dynamics of pollen tube growth in pistil, and yield and other fruiting attributes, in 'Satluj Purple' and 'Kala Amritsari', probably the most popular subtropical Japanese plum varieties in northern regions of India. Specifically, we examined the response of six different pollination variants, namely to self-pollination, open-pollination with the two cultivars located in adjacent rows, open-pollination with the two cultivars located in distant rows, manual cross-pollination, supplementary pollination, and floral bouquet. During the two years of the investigation, both plum cultivars showed good in vitro pollen germination (on average, above 50%) at different sucrose concentrations, with the highest values for the 'Satluj Purple' and for the 15% concentration. In vivo, the analysis of the pollen growth in the various sections of the style indicated the best performance when pistils of 'Satluj Purple' were pollinated by pollen grains of cv. 'Kala Amritsari'. Cross-pollination also registered faster growth of pollen tube in pistil with the lowest number of incompatible pollen tubes compared to open- and self-pollination. From the productive point of view, cross-pollination showed the most pronounced results among the different pollination variants, with the highest initial fruit set (36.6%) and yield (28.0 kg/tree), and the shorter fruit development in 'Satluj Purple' (fruit set and yield in self-pollinated 'Satluj Purple' trees were 3.3% and 2.0 kg/tree, respectively). Conversely, the use of 'Satluj Purple' pollen for 'Kala Amritsari' showed poor results. Finally, in our study, 'Kala Amritsari' showed self-compatibility. We conclude that the main cause of poor fruit set in 'Satluj Purple' is self-incompatibility. The relevant genotypic-specific effects revealed by the analysis of the various pollination treatments also highlighted the importance of interplanting to increase fruit set and yield for subtropical Japanese plum varieties. [ABSTRACT FROM AUTHOR]

In Brassicaceae, the papillary cells of the stigma are the primary site of the self-incompatibility (SI) responses. SI preserves the genetic diversity by selectively rejecting irrelevant or incompatible pollen, thus promoting cross fertilization and species fitness. Mechanisms that regulate SI responses in Brassica have been studied mainly on the mature stigma that often undermines how stigma papillary cells attain the state of SI during development. To understand this, we integrated PacBio SMRT-seq with Illumina RNA-seq to construct a de novo full-length transcriptomic database for different stages of stigma development in ornamental kale. A total of 48,800 non-redundant transcripts, 31,269 novel transcripts, 24,015 genes, 13,390 alternative splicing, 22,389 simple sequence repeats, 21,816 complete ORF sequences, and 4591 lncRNAs were identified and analyzed using PacBio SMRT-seq. The Illumina RNA-seq revealed 15,712 differentially expressed genes (DEGs) and 8619 transcription factors. The KEGG enrichment analysis of 4038 DEGs in the "incompatibility" group revealed that the flavonoid and fatty acid biosynthesis pathways were significantly enriched. The cluster and qRT-PCR analysis indicated that 11 and 14 candidate genes for the flavonoid and fatty acid biosynthesis pathways have the lowest expression levels at stigma maturation, respectively. To understand the physiological relevance of the downregulation of fatty acid biosynthesis pathways, we performed inhibitor feeding assays on the mature stigma. The compatible pollination response was drastically reduced when mature stigmas were pre-treated with a fatty acid synthase inhibitor. This finding suggested that fatty acid accumulation in the stigmas may be essential for compatible pollination and its downregulation during maturity must have evolved as a support module to discourage the mounting of self-incompatible pollen. [ABSTRACT FROM AUTHOR]

Self-incompatibility (SI) responses of radish (Raphanus sativus L.) are determined by two tightly linked genes encoding an S receptor kinase (SRK) and an S-locus cysteine-rich protein/S locus protein 11 (SCR/SP11), respectively. A radish showing an almost self-compatible (SC) phenotype was identified in this study. Inheritance patterns showed that this SC phenotype was dominant over an SI phenotype. In addition, the SC phenotype co-segregated with an S haplotype in an F2 population. This SC radish contained an RsS-26 haplotype in which duplicate SRK-like genes were previously identified. Full-length sequences of two SRK-like genes of 18,133-bp and 6,200-bp in length were obtained, and they were designated as RsSRK-26-1 and RsSRK-26-2, respectively. No critical mutation was found in the coding region of SRK-like genes. However, a 4,146-bp intact LTR-retrotransposon was identified in the third intron of RsSRK-26-1 of the SC radish. Interestingly, this LTR-retrotransposon was not detected in three other breeding lines containing the same RsS-26 haplotype. While normal transcription of RsSRK-26-1 and SI phenotypes were observed in these three breeding lines, no transcript of RsSRK-26-1 was detected in the SC radish, suggesting that recent transposition of an LTR-retrotransposon in the RsSRK-26-1 gene might be responsible for the SC phenotype of radish. [ABSTRACT FROM AUTHOR]

Self-incompatibility (SI) is a major issue in dragon fruit (Selenicereus spp.) breeding and production. Therefore, a better understanding of the dragon fruit SI mechanism is needed to improve breeding efficiency and ultimate production costs. To reveal the underlying mechanisms of SI in dragon fruit, plant anatomy, de novo RNA sequencing-based transcriptomic analysis, and multiple bioinformatic approaches were used to analyze gene expression in the pistils of the self-pollinated and cross-pollinated dragon fruit flowers at different intervals of time after pollination. Using fluorescence microscopy, we observed that the pollen of 'Hongshuijing', a self-incompatible dragon fruit variety (S. monacanthus), germinated on its own stigma. However, the pollen tube elongation has ceased at 1/2 of the style, confirming that dragon fruit experiences gametophyte self-incompatibility (GSI). We found that the pollen tube elongation in vitro was inhibited by self-style glycoproteins in the SI variety, indicating that glycoproteins were involved in SI. That is to say the female S factor should be homologous of S-RNase or PrsS (P. rhoeas stigma S factor), both of which are glycoproteins and are the female S factors of the two known GSI mechanism respectively. Bioinformatics analyses indicated that among the 43,954 assembled unigenes from pistil, there were six S-RNase genes, while 158 F-box genes were identified from a pollen transcriptomic dataset. There were no P. rhoeas type S genes discovered. Thus, the identified S-RNase and F-box represent the candidate female and male S genes, respectively. Analysis of differentially expressed genes (DEGs) between the self and cross-pollinated pistils at different time intervals led to the identification of 6,353 genes. We then used a weighted gene co-expression network analysis (WGCNA) to find some non-S locus genes in SI responses in dragon fruit. Additionally, 13 transcription factors (TFs) (YABBY4, ANL2, ERF43, ARF2, BLH7, KNAT6, PIF3, two OBF1, two HY5 and two LHY/CCA) were identified to be involved in dragon fruit GSI. Thus, we uncovered candidate S and non-S genes and predicted more SI-related genes for a more detailed investigation of the molecular mechanism of dragon fruit SI. Our findings suggest that dragon fruit possesses a GSI system and involves some unique regulators. This study lays the groundwork for future research into SI mechanisms in dragon fruit and other plant species. [ABSTRACT FROM AUTHOR]

Self-incompatibility (SI) is a feature of many flowering plants, whereby self-pollen is recognized and rejected by the stigma. In grasses (Poaceae), the genes controlling this phenomenon have not been fully elucidated. Grasses have a unique two-locus system, in which two independent genetic loci (S and Z) control self-recognition. S and Z are thought to have arisen from an ancient duplication, common to all grasses. With new chromosome-scale genome data, we examined the genes present at S- and Z-loci, firstly in ryegrass (Lolium perenne), and subsequently in ~20 other grass species. We found that two DUF247 genes and a short unstructured protein (SP/ZP) were present at both S- and Z- in all SI species, while in self-compatible species these genes were often lost or mutated. Expression data suggested that DUF247 genes acted as the male components and SP/ZP were the female components. Consistent with their role in distinguishing self- from non-self, all genes were hypervariable, although key secondary structure features were conserved, including the predicted N-terminal cleavage site of SP/ZP. The evolutionary history of these genes was probed, revealing that specificity groups at the Z-locus arose before the advent of various grass subfamilies/species, while specificity groups at the S-locus arose after the split of Panicoideae, Chloridoideae, Oryzoideae and Pooideae. Finally, we propose a model explaining how the proteins encoded at the S and Z loci might function to specify self-incompatibility. [ABSTRACT FROM AUTHOR]

In heterostylous, self-incompatible Turnera species, a member of the YUCCA gene family, YUC6, resides at the S-locus and has been hypothesized to determine the male mating type. YUCCA gene family members synthesize the auxin, indole-3-acetic acid, via a two-step process involving the TAA gene family. Consequently, it has been speculated that differences in auxin concentration in developing anthers are the biochemical basis underlying the male mating type. Here, we provide empirical evidence that supports this hypothesis. Using a transgenic knockdown approach, we show that YUC6 acts pleiotropically to control both the male physiological mating type and pollen size, but not the filament length dimorphism associated with heterostyly in Turnera. Using qPCR to assess YUC6 expression in different transgenic lines, we demonstrate that the level of YUC6 knockdown correlates with the degree of change observed in the male mating type. Further assessment of YUC6 expression through anther development, in the knockdown lines, suggests that the male mating type is irreversibly determined during a specific developmental window prior to microsporogenesis, which is consistent with the genetically sporophytic nature of this self-incompatibility system. These results represent the first gene controlling male mating type to be characterized in any species with heterostyly. [ABSTRACT FROM AUTHOR]

Summary: Ascidians (Urochordate) are hermaphroditic marine invertebrates that release sperm and eggs to the surrounding seawater. However, several ascidians, including Ciona intestinalis and Halocynthia roretzi, show strict self-sterility due to a self/nonself-recognition mechanism in the interaction between sperm and the vitelline coat (VC) of the eggs. We have previously reported that sperm intracellular Ca2+ level drastically increased immediately after sperm binding to the VC of self eggs but not nonself eggs in C. intestinalis type A, which was potently inhibited by lowering the external Ca2+ concentration, suggesting that sperm Ca2+ influx occurs after sperm self-recognition on the VC. Here, we investigated whether self-sterility was abolished by lowering the external Ca2+ concentration in C. intestinalis. The results showed that the block to self-fertilization was removed by low-Ca2+ (∼1 mM) seawater without decreasing the fertilization rate. Such an effect was not observed with Mg2+ or K+. These results led us to conclude that a low-Ca2+ environment is sufficient to block the self-recognition signal upon fertilization. As low-Ca2+ seawater showed no effect on H. roretzi self-sterility, we propose that the mechanism of self-sterility in Ciona must be distinctive from that in Halocynthia. [ABSTRACT FROM AUTHOR]

S-RNase plays vital roles in the process of self-incompatibility (SI) in Rutaceae plants. Data have shown that the rejection phenomenon during self-pollination is due to the degradation of pollen tube RNA by S-RNase. The cytoskeleton microfilaments of pollen tubes are destroyed, and other components cannot extend downwards from the stigma and, ultimately, cannot reach the ovary to complete fertilisation. In this study, four S-RNase gene sequences were identified from the 'XiangShui' lemon genome and ubiquitome. Sequence analysis revealed that the conserved RNase T2 domains within S-RNases in 'XiangShui' lemon are the same as those within other species. Expression pattern analysis revealed that S3-RNase and S4-RNase are specifically expressed in the pistils, and spatiotemporal expression analysis showed that the S3-RNase expression levels in the stigmas, styles and ovaries were significantly higher after self-pollination than after cross-pollination. Subcellular localisation analysis showed that the S1-RNase, S2-RNase, S3-RNase and S4-RNase were found to be expressed in the nucleus according to laser confocal microscopy. In addition, yeast two-hybrid (Y2H) assays showed that S3-RNase interacted with F-box, Bifunctional fucokinase/fucose pyrophosphorylase (FKGP), aspartic proteinase A1, RRP46, pectinesterase/pectinesterase inhibitor 51 (PME51), phospholipid:diacylglycerol acyltransferase 1 (PDAT1), gibberellin receptor GID1B, GDT1-like protein 4, putative invertase inhibitor, tRNA ligase, PAP15, PAE8, TIM14-2, PGIP1 and p24beta2. Moreover, S3-RNase interacted with TOPP4. Therefore, S3-RNase may play an important role in the SI of 'XiangShui' lemon. [ABSTRACT FROM AUTHOR]

Fragaria viridis exhibits S-RNase-based gametophytic self-incompatibility, in which S-RNase is the major factor inhibiting pollen tube growth. However, the pathways involved in and the immediate causes of the inhibition of pollen tube growth remain unknown. Here, interactive RNA sequencing and proteome analysis revealed changes in the transcriptomic and proteomic profiles of F. viridis styles harvested at 0 and 24 h after self-pollination. A total of 2,181 differentially expressed genes and 200 differentially abundant proteins were identified during the pollen development stage of self-pollination. Differentially expressed genes and differentially abundant proteins associated with self-incompatible pollination were further mined, and multiple pathways were found to be involved. Interestingly, the expression pattern of the transcription factor FviYABBY1, which is linked to polar growth, differed from those of other genes within the same family. Specifically, FviYABBY1 expression was extremely high in pollen, and its expression trend in self-pollinated styles was consistent with that of S-RNase. Furthermore, FviYABBY1 interacted with S-RNase in a non-S haplotype way. Therefore, FviYABBY1 affects the expression of polar growth-related genes in self-pollen tubes and is positively regulated by S-RNase. [ABSTRACT FROM AUTHOR]

Breakdown of self‐incompatibility increases opportunities for both self‐fertilization and interspecific hybridization, although the latter is dependent on the extent of competition between heterospecific and conspecific pollen. We investigate the mating system and pollination biology of five phylogenetically closely related species within a distylous species complex in Hedyotis L. (Rubiaceae) in southern China. The complex comprises Hedyotis acutangula Champ. ex Benth., Hedyotis shiuyingiae T.Chen, Hedyotis vachellii (Hook. & Arn.) Kuntze, and two putative hybrid species, Hedyotis bodinieri (H.Lév.) Chun and Hedyotis loganioides Benth., hypothesized to result from interbreeding between these species. We test the hypothesis that the breakdown of self‐ and interspecific incompatibilities in sympatric Hedyotis species might allow interspecific hybridization in natural populations. We assessed the extent of self‐ and interspecific incompatibility in sympatric populations, including investigations of spontaneous and artificial self‐pollination, geitonogamy, inter‐ and intramorph xenogamy. Artificial interspecific crosses were undertaken between H. acutangula, H. shiuyingiae, and H. bodinieri, between H. acutangula and H. vachellii, and between H. acutangula and H. loganioides. Hedyotis acutangula is demonstrated to be self‐ and interspecific compatible, whereas H. vachellii, H. bodinieri, and H. loganioides are self‐compatible and interspecific incompatible; H. shiuyingiae, in contrast, is strictly self‐ and interspecific incompatible. Comparisons of pollen tube growth rates in hybridizing species‐pairs revealed that heterospecific pollen of H. shiuyingiae, H. vachellii, and H. bodinieri can compete with conspecific self‐pollen of H. acutangula. Our study therefore indicates that the breakdown of self‐incompatibility directly and indirectly facilitates interspecific hybridization and provides a platform for better understanding evolutionary directionality in Hedyotis. [ABSTRACT FROM AUTHOR]

Radish is a typical self-incompatible crop. The rapid and accurate identification of S haplotypes can circumvent the blindness of the hybrid combination process, which is critical in radish heterosis utilization and the breeding of new varieties. In this study, based on the gene sequence which encodes the S-locus receptor kinase (SRK) of radish, and the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, the S haplotypes were identified among 79 cultivated radish genotypes. The PCR results indicated that 79 radish genotypes could be divided into 48 Class I, 13 Class II, and 17 Class I/II S haplotypes. Sequence alignment confirmed that the Class I materials contained 19 S haplotypes, of which three haplotypes ('NAU-S53', 'NAU-S54' and 'NAU-S55') were identified for the first time in radish. After digestion using the Hinf I restriction endonuclease, the SRK domain of DNA fragments of different genotypes showed high polymorphism. Homozygous materials S haplotypes could be quickly distinguished by the differences in the digested bands. Molecular identification of the S haplotype was highly consistent with the field pollination and pollen tube germination results. These results would provide an important approach for the rapid identification of radish S haplotypes and the efficient utilization of self-incompatibility in heterosis breeding. [ABSTRACT FROM AUTHOR]

Self-incompatibility (SI) is a common strategy to avoid inbreeding and, consequently, keep genetic diversity within a species. In its mechanism, pollen rejection happens in the style when the single multiallelic locus (SFB in prunus species) of the haploid pollen matches one of the S-alleles existing in the diploid pistil. The SFB gene for the pollen S gene has been identified in many Prunus species. However, Japanese apricot is a species with a typical gametophytic self-incompatibility (GSI), and its SFB alleles available are limited, although they are required for studying GSI. Therefore, we used an AS-PCR amplification method, sequencing, and the pair primers SFB-C1F and Pm-Vb designed based on the conserved region of the Prunus SFB gene to identify SFB genotypes of 48 Japanese apricot (P. mume) accessions. Eleven novel SFB alleles were isolated from these accessions and shared typical structural features with SFB alleles from other Prunus species. These novel SFB alleles were uniquely expressed in pollen. Hence, we concluded that these 11 PmSFB were pollen S determinants of P. mume. This current study offers the novel SFB genes of the P. mume S locus, which could be a useful potential resource for studies on pollen SI mechanisms. [ABSTRACT FROM AUTHOR]

Habitat fragmentation and the acceleration of environmental change threaten the survival of many plant species. The problem is especially pronounced for plant species with self-incompatibility mating systems, which are obligate outcrossers, thus requiring high mate availability to persist. In such situations, plant populations suffering decreased fitness could be rescued by: (a) improving local habitat conditions (habitat rescue), (b) increasing the number of individuals (demographic rescue), or (c) introducing new genetic variation (genetic rescue). In this study, we used a spatially and genetically explicit individual-based model to approximate the demography of a small ( N  = 250) isolated self-incompatible population using a timescale of 500 years. Using this model, we quantified the effectiveness of the different types of rescues described above, singly and in combination. Our results show that individual genetic rescue is the most effective type of rescue with respect to improving fitness and population viability. However, we found that introducing a high number of individuals ( N  > 30) to a small population ( N  = 50) at the brink of extinction through demographic rescue can also have a positive effect on viability, improving average fitness by 55% compared to introducing a low number of individuals ( N  = 10) over a long timescale (> 500 years). By itself, habitat rescue showed the lowest effects on viability. However, combining genetic and habitat rescue provided the best results overall, increasing both persistence (> 30%) and mate availability (> 50%). Interestingly, we found that the addition of even a small number of new S alleles (20%) can be highly beneficial to increase mate availability and persistence. We conclude that genetic rescue through the introduction of new S alleles and an increase in habitat suitability is the best management strategy to improve mate availability and population viability of small isolated SI plant populations to overcome the effects of demographic stochasticity and positive density dependence., Competing Interests: The authors declare no conflicts of interest., (© 2024 Commonwealth Scientific and Industrial Research Organisation. Evolutionary Applications published by John Wiley & Sons Ltd.)

The plant elongation factor eEF1A is involved in coregulating not only the translation of proteins and controlling translation-related signaling but also in signaling associated with cell growth, stress response and motility, controlling apoptosis and responding to adversity in plants. In this study, four eEF1A genes, namely, ClEF1A-1, ClEF1A-2, ClEF1A-3 and ClEF1A-4, were identified from the genomic and ubiquitin-modified omics data of the 'Xiangshui Lemon', and bioinformatics analysis revealed that these four genes have relatively similar structures with conserved sequences; ClEF1A-1 and ClEF1A-4 were highly expressed in pollen, and temporal expression analysis demonstrated that the expression of ClEF1As was significantly greater under self-pollination than under cross-pollination. All four genes were localized in the nucleus. ClEF1As overexpression promoted early flowering and improved drought and salt stress tolerance in transgenic Arabidopsis plants. Yeast two-hybrid assays revealed that ClEF1As interacted with F-box, eIF3-G, the organ-specific-like protein S2, AGL62, S 1 -RNase, S 2 -RNase, S 3 -RNase and S 4 -RNase. This study demonstrated the functions of ClEF1As and provided a baseline for further studies on the associations of ClEF1As with self-incompatibility and abiotic stresses., Competing Interests: Declaration of competing interest We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled “Four ClEF1A genes involved in self-incompatibility in 'Xiangshui Lemon' confer early fowering and increase stress tolerance in transgenic Arabidopsis”. All authors have seen the manuscript and approved the submittal to your journal. None of the material in the paper has been published or is under consideration for publication elsewhere., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)