Correlation of self- and interspecific incompatibility among sympatric Hedyotis species (Rubiaceae) and consequences for hybridization

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.     

Incompatibility in Dendrobium (Orchidaceae)

JOHANSEN, B., 1990. Incompatibility in Dendrobium (Orchidaceae). A unique self-incompatibility system in Dendrobium is demonstrated by more than 1700 pollination experiments. The majority (72%) of the 61 species that were self-pollinated showed self-sterility. In contrast with many other orchid genera Dendrobium showed high incompatibility in interspecific pollinations. Self-and interspecific incompatibility is expressed by flower abscission and not by inhibition of pollen germination or pollen tube growth. The incompatibility system is gametophytic and complementary, and it is likely that the auxin content in the pollinia triggers the incompatibility reaction. Microscopical investigations on the detached cells of the stigma (here called eleutherocytes) after compatible and incompatible pollinations, suggest that the incompatibility response is probably controlled by these cells.     

Frequency of interspecific matings and of hybrid offspring in sympatric populations of Vtviparus ater and V. contectus (Mollusca: Prosobranchia)

In species pairs able to produce fertile hybrids, the amount of introgression can be limited by the frequency of interspecific copulations. We investigated the frequency of interspecific copulations between Kviparus ater and V. contectus and the proportion of hybrid offspring of V. ater females in natural populations in northern Italy. Although there was significant assortative mating, the frequency of interspecific copulations was substantial. At sites where V. contectus was rare, 5.9% of all matings, and up to 73% of matings in which V. contectus was involved, were interspecific. At sites where both species were abundant 14.8% of all matings were interspecific. Male-male matings occurred in approximately 3% of copulations. In 1993, 12 out of 325 V. ater females collected from 10 sympatric populations and held in isolation for two to five months gave birth to 25 hybrid offspring, out of a total of 2548 offspring produced by all females. The proportion of hybrid offspring (ranging from 0 to 2.3% depending on the site) was positively related to the proportion of V. contectus in the population. Since interspecific copulations are frequent and hybrid offspring are produced in nature, we conclude that assortative mating is not an effective isolating mechanism between V. ater and V. contectus.     

A new look at incompatibility relationships in higher plants

Summary Pollen tube growth was evaluated using an 18-step scale after both intra- and interspecific pollinations of genotypically widely differing diploid potato species and potato dihaploids expressing monofactorial gametophytic incompatibility. The results obtained account for a wide array of types of pollen tube growth resulting from crossing partners with distinct incompatibility behavior. Based on the assumption that inhibition of pollen tubes is the rule and solely prevented by the pollen itself, a model proposing one common cause underlying different mechanisms for both intraspecific selfincompatibility and interspecific incompatibility in diploids is put forward. Data supporting the model are presented from the experimental results of this study and from the literature. The strength of pollen-style interaction depends on particular S-alleles in combination with recognition and activity properties of the basic S-genotypes. The model is suitable to explain all former observations on incompatibility in diploids with a gametophytic system of incompatibility and, with modifications of the manner of phenotypic expression, also in plants with sporophytic incompatibility. The proposed scheme of pollen tube growth phenotypes permits prediction of pollen tube growth behavior in an intended cross combination. The model is based on both classical Mendelian genetics and recent molecular genetic insight.     

Unilateral incongruity in crosses involvingLycopersicon pennellii andL. esculentum is distinct from self-incompatibility in expression,timing and location

Both interspecific and intraspecific mechanisms restrict the exchange of genes between plants. Much research has focused on self incompatibility (SI), an intraspecific barrier, but research on interspecific barriers lags behind. We are using crosses betweenLycopersicon esculentum andL. pennellii as a model with which to study interspecific crossing barriers. The crossL. esculentum×L. pennellii is successful, but the reciprocal cross fails. Since the cross can be successfully made in one direction but not the other, gross genomic imbalance or chromosomal abnormality are precluded as causes. We showed that the lack of seed set observed in the crossL. pennellii×L. esculentum is due to the inability of pollen tubes to grow more than 2–3 mm into the style, whereas S1 crosses show continued slow pollen tube growth but, also, fail to set seed. These results indicate that the unilateral response is a barrier distinct from SI, differing from SI in the timing and location of expression in the style. We therefore suggest that this unilateral response in theL. pennellii×L. esculentum cross is more accurately referred to as unilateral incongruity (UI) rather than interspecific incompatibility. Periclinal chimeras were used to determine the tissues involved in UI. The results of crosses with the available chimeras indicate that the female parent must beL. pennellii at either LI (layer 1) or both LI and LII (layer 2) and the male parent must beL. esculentum at either LII or both LI and LII to observe UI similar to that seen in theL. pennellii×L. esculentum cross. Pollinations with a mixture of pollen fromL. pennellii and from transgenicL. esculentum plants harboring a pollen-specific GUS reporter gene marker were used to ascertain whether the growth of the pollen tubes of either species was modified as a possible means of overcoming UI. We found no evidence of communication between the two types of pollen tubes to either enhance or restrict all pollen tube growth.     

Reproductive systems and low outbreeding barriers between Jacaranda cuspidifolia and J. mimosifolia (Jacarandeae,Bignoniaceae)

The genus Jacaranda shows notable karyotype stability and a prevailing self‐sterile breeding system with evidence of late‐acting self‐incompatibility in several species. However, some studies have indicated self‐compatibility in J. mimosifolia, a species cultivated worldwide in tropical and subtropical areas. Jacaranda cuspidifolia is a closely related species with natural distribution broadly overlapping with that of J. mimosifolia, and manual heterospecific pollination studies have indicated that these species are interfertile, but there is no report on the breeding system of the former. In this study, we used hand‐pollination experiments, pistil longevity, epi‐fluorescence and histological analysis of post‐pollination events to determine the breeding system of J. cuspidifolia. We also employed intra and interspecific crosses and seed germination tests to reevaluate the breeding system of J. mimosifolia and the inter‐fertility between the two species. Some fruits were initiated from self‐pollinated pistils in J. mimosifolia, but none of them reached maturity. On the other hand, complete absence of fruit development by self‐pollination was verified in J. cuspidifolia, while in situ pollen tube growth and histological analysis of post‐pollination events in selfed pistils revealed the characteristic ovule penetration, fertilization and endosperm initiation also observed in other bignoniaceous species with late‐acting self‐incompatibility. Low outbreeding barriers seem to operate between these species because reciprocal interspecific crosses and hybrid seed germination tests indicated they are bilaterally interfertile. However, fruit/seed production and seed germinability were significantly lower when pistils of J. cuspidifolia were pollinated with pollen of J. mimosifolia, compared with crosses in the opposite direction, which indicates a partial unilateral incompatibility. This result is discussed in the context of the possible occurrence of self‐compatibility in J. mimosifolia. The low level of incongruity operating between the two species also points to their recent evolutionary divergence.     

GENOME‐WIDE INVESTIGATION OF REPRODUCTIVE ISOLATION IN EXPERIMENTAL LINEAGES AND NATURAL SPECIES OF NEUROSPORA: IDENTIFYING CANDIDATE REGIONS BY MICROARRAY‐BASED GENOTYPING AND MAPPING

Inherent incompatibilities between genetic components from genomes of different species may cause intrinsic reproductive isolation. In evolution experiments designed to instigate speciation in laboratory populations of the filamentous fungus Neurospora, we previously discovered a pair of incompatibility loci (dfe and dma) that interact negatively to cause severe defects in sexual reproduction. Here we show that the dfe‐dma incompatibility also is a significant cause of genetic isolation between two naturally occurring species of Neurospora (N. crassa and N. intermedia). The strong incompatibility interaction has a simple genetic basis (two biallelic loci) and antagonistic epistasis occurs between heterospecific alleles only, consistent with the Dobzhansky–Muller model of genic incompatibility. We developed microarray‐based, restriction‐site associated DNA (RAD) markers that identified ～1500 polymorphisms between the genomes of the two species, and constructed the first interspecific physical map of Neurospora. With this new mapping resource, the approximate genomic locations of the incompatibility loci were determined using three different approaches: genome scanning, bulk‐segregant analyses, and introgression. These population, quantitative, and classical genetics methods concordantly identified two candidate regions, narrowing the search for each incompatibility locus to only ～2% of the nuclear genome. This study demonstrates how advances in high‐throughput, genome‐wide genotyping can be applied to mapping reproductive isolation genes and speciation research.     

Albinism in Plants: A Major Bottleneck in Wide Hybridization,Androgenesis and Doubled Haploid Culture

Albinism is a common problem encountered in interspecific crosses and tissue culture experiments including anther culture and generation of doubled haploids. It is characterized by partial or complete loss of chlorophyll pigments and incomplete differentiation of chloroplast membranes. This in turn impairs photosynthesis and the plants eventually die at a young stage without reaching maturity. Environmental conditions such as light, temperature, media composition and culture conditions play some role in determining the frequency of albino plant formation. Genetic factors are even more important, and are major determinants in albinism. Genetic studies in different crops show that it is a recessive trait governed by many loci. Both the nuclear and chloroplast genomes affect albinism and incompatibilities between the two are a probable cause of many pigment defects in hybrid progenies. Such incompatibility has been reported in a large number of angiosperms. The mechanisms behind these incompatibilities are poorly understood. Studies of plastid DNA inheritance together with observations using electron microscopy have established that the transmission of plastids can be maternal, paternal or biparental, even within the same genus, especially following wide crosses; contrary to the widespread belief that plastids are almost always transmitted from the maternal parent. Albinism has been overcome in some crop species through somatic hybridization and development of cybrids (cytoplasmic hybrids). However, the strict requirement of efficient protoplast regeneration is a major limitation of these techniques. This review focuses on albinism following interspecific crosses or development of doubled haploids facilitated by tissue culture experiments, underlying mechanisms, and the possibilities for dealing with this important biotechnological limitation.     

Unilateral incompatibility in Capsicum (Solanaceae): occurrence and taxonomic distribution

BACKGROUND AND AIMS: Unilateral incompatibility (UI) occurs when pollinations between species are successful in one direction but not in the other. Self-incompatible (SI) species frequently show UI with genetically related, self-compatible (SC) species, as pollen of SI species is compatible on the SC pistil, but not vice versa. Many examples of unilateral incompatibility, and all those which have been studied most intensively, are found in the Solanaceae, particularly Lycopersicon, Solanum, Nicotiana and Petunia. The genus Capsicum is evolutionarily somewhat distant from Lycopersicon and Solanum and even further removed from Nicotiana and Petunia. Unilateral incompatibility has also been reported in Capsicum; however, this is the first comprehensive study of crosses between all readily available species in the genus. METHODS: All readily available (wild and domesticated) species in the genus are used as plant material, including the three genera from the Capsicum pubescens complex plus eight other species. Pollinations were made on pot-grown plants in a glasshouse. The number of pistils pollinated per cross varied (from five to 40 pistils per plant), depending on the numbers of flowers available. Pistils were collected 24 h after pollination and fixed for 3-24 h. After staining, pistils were mounted in a drop of stain, squashed gently under a cover slip and examined microscopically under ultra-violet light for pollen tube growth. KEY RESULTS: Unilateral incompatibility is confirmed in the C. pubescens complex. Its direction conforms to that predominant in the Solanaceae and other families, i.e. pistils of self-incompatible species, or self-compatible taxa closely related to self-incompatible species, inhibit pollen tubes of self-compatible species. CONCLUSIONS: Unilateral incompatibility in Capsicum does not seem to have arisen to prevent introgression of self-compatibility into self-incompatible taxa, but as a by-product of divergence of the C. pubescens complex from the remainder of the genus.     

Interspecific incompatibility due to stylar barriers in tuber-bearing and closely related non-tuber-bearing Solanums

Summary The phenomenon of interspecific incompatibility between various wild tuber-bearing and closely related non-tuber-bearing Solanum species was studied. One area of investigation included an examination of possible protein interactions in the incompatibility reaction using SDS electrophoresis. Pollen tube inhibition and morphology were examined in conjunction with biochemical analysis. Two sets of crosses were examined: interspecific tuber-bearing species crosses and interspecific tuber-bearing × non-tuber-bearing species crosses. These crosses had consistent pollen tube inhibition in the upper one-third of the style. The upper third of the styles of incompatibly pollinated, compatibly pollinated, and unpollinated styles was studied under fluorescence microscopy to observe pollen tube growth and morphology. Interspecific tuber-bearing × non-tuber-bearing species crosses demonstrated consistent pollen tube inhibition just below the stigma with frequent pollen tube swelling and bursting and extensive callose deposition along the pollen tube wall. Interspecific tuber-bearing species crosses had pollen tube inhibition further down the style with pollen tube tip tapering and extensive callose deposition. Stylar proteins of the lower two-thirds of the styles were analyzed with SDS electrophoresis. No unique protein differences were found to be specifically associated with the interspecific incompatibility reaction in this portion of the style.Cooperative investigation of the U.S. Department of Agriculture, Agricultural Research Service, and the Wisconsin Experiment Station. Supported in part by the USDA, Cooperative States Research Service Competitive grant no. 83-CRCR-1-1253     

崖柏群落优势乔木树种种间关系

采用样方调查法,对组成崖柏群落的乔木树种进行调查。通过计算重要值确定崖柏群落优势乔木树种,研究优势树种的总体联结性、2物种间的联结性和种间协变。结果表明:崖柏群落内的15个优势树种总体间存在负关联;2物种间具有显著正联结的种对有:高山栎-川陕鹅耳枥、高山栎-华中八角、大叶青冈-乌岗栎、铁杉-华西花楸、华千金榆-青榨槭、华千金榆-大叶青冈、大叶青冈-川鄂山茱萸、川陕鹅耳枥-华中八角;呈显著负联结的种对是高山栎-青榨槭;崖柏与其他优势树种的联结性均未达到显著程度;有26个种对表现出明显的正协变,12个种对表现出明显的负协变;种对正协变的存在是由于这些物种对环境资源的利用具有相似性所致。     

高等担子菌的性非亲和性系统和体细胞非亲和性系统在遗传多样性保存中的作用

本文讨论了高等担子菌的个体概念,以及利用性的非亲和性、体细胞非亲和性对于高等担子菌个体的识别,非亲和性系统对于真菌本身在遗传多样性的保存中的作用。担子菌的个体概念对于其生态学研究具有很大的意义,控制担子菌性非亲和性的交配型基因可以作为遗传多样性研究的重要手段。     

Natural selection reinforces speciation in a radiation of neotropical rainforest plants

The importance of reinforcement, that is, natural selection that strengthens reproductive isolation between incipient species, remains controversial. We used two approaches to test for reinforcement in a species radiation of Neotropical gingers in the genus Costus. First, we conducted an intensive study of Costus pulverulentus and Costus scaber, two recently diverged species that co-occur and share hummingbird pollinators. The hummingbird pollinators transfer pollen between these Costus species, but hybrids are rarely found in nature. By performing pollinations between populations of C. pulverulentus and C. scaber from three sites across the species' geographic ranges, we find that pollen-pistil incompatibilities acting prior to fertilization have evolved only between locally sympatric populations, whereas geographically distant populations within the region of sympatry and allopatric populations remain fully interfertile. Second, we conducted a comparative study of isolating mechanisms across the genus. We find lower seed set due to pollen-pistil incompatibility between species pairs that co-occur and experience pollen transfer in nature compared to species pairs that are otherwise isolated, regardless of genetic distance. Taken together, these studies indicate that crossing barriers prevent potentially maladaptive hybridization and effectively reinforce the speciation process. Our results add to mounting evidence for reinforcement from animal studies and show that plant speciation may also involve complex mate recognition systems. Reinforcement may be particularly important in rapidly diverging lineages where ecological factors play a primary role in reproductive isolation, as may often be the case in tropical communities.     

Austrobilharzia terrigalensis: A schistosome dominant in interspecific interactions in the molluscan host

Walker J. C. 1979. Austrobilharzia terrigalensis: a schistosome dominant in interspecific interactions in the molluscan host. International Journal for Parasitology9: 137–140. In the estuarine prosobranch Velacumantus australis the avian schistosome Austrobilharzia terrigalensis is always associated with the germinal sacs of other trematodes. In this association the schistosome retards the development of the other trematode, thereby reducing the intensity of interspecific competition for the limited resource, the host's tissues.     

Breeding systems in Cyrtanthus (Amaryllidaceae): variation in self‐sterility and potential for ovule discounting

• Breeding systems of plants determine their reliance on pollinators and ability to produce seeds following self‐pollination. Self‐sterility, where ovules that are penetrated by self‐pollen tubes that do not develop into seeds, is usually considered to represent either a system of late‐acting self‐incompatibility or strong early inbreeding depression. Importantly, it can lead to impaired female function through ovule or seed discounting when stigmas receive mixtures of self and cross pollen, unless cross pollen is able to reach the ovary ahead of self pollen (‘prepotency’). Self‐sterility associated with ovule penetration by self‐pollen tubes appears to be widespread among the Amaryllidaceae.
• We tested for self‐sterility in three Cyrtanthus species – C. contractus, C. ventricosus and C. mackenii – by means of controlled hand‐pollination experiments. To determine the growth rates and frequency of ovule penetration by self‐ versus cross‐pollen tubes, we used fluorescence microscopy to examine flowers of C. contractus harvested 24, 48 and 72 h after pollination, in conjunction with a novel method of processing these images digitally. To test the potential for ovule discounting (loss of cross‐fertilisation opportunities when ovules are disabled by self‐pollination), we pollinated flowers of C. contractus and C. mackenii with mixtures of self‐ and cross pollen.
• We recorded full self‐sterility for C. contractus and C. ventricosus, and partial self‐sterility for C. mackenii. In C. contractus, we found no differences in the growth rates of self‐ and cross‐pollen tubes, nor in the proportions of ovules penetrated by self‐ and cross‐pollen tubes. In this species, seed set was depressed (relative to cross‐pollinated controls) when flowers received a mixture of self and cross pollen, but this was not the case for C. mackenii.
• These results reveal variation in breeding systems among Cyrtanthus species and highlight the potential for gender conflict in self‐sterile species in which ovules are penetrated and disabled by pollen tubes from self pollen.

     

Natural variation for a hybrid incompatibility between two species of Mimulus

Understanding the process by which hybrid incompatibility alleles become established in natural populations remains a major challenge to evolutionary biology. Previously, we discovered a two-locus Dobzhansky-Muller incompatibility that causes severe hybrid male sterility between two inbred lines of the incompletely isolated wildflower species, Mimulus guttatus and M. nasutus. An interspecific cross between these two inbred lines revealed that the M. guttatus (IM62) allele at hybrid male sterility 1 (hms1) acts dominantly in combination with recessive M. nasutus (SF5) alleles at hybrid male sterility 2 (hms2) to cause nearly complete hybrid male sterility. In this report, we extend these genetic analyses to investigate intraspecific variation for the hms1-hms2 incompatibility in natural populations of M. nasutus and M. guttatus, performing a series of interspecific crosses between individuals collected from a variety of geographic locales. Our results suggest that hms2 incompatibility alleles are common and geographically widespread within M. nasutus, but absent or rare in M. guttatus. In contrast, the hms1 locus is polymorphic within M. guttatus and the incompatibility allele appears to be extremely geographically restricted. We found evidence for the presence of the hms1 incompatibility allele in only two M. guttatus populations that exist within a few kilometers of each other. The restricted distribution of the hms1 incompatibility allele might currently limit the potential for the hms1-hms2 incompatibility to act as a species barrier between sympatric populations of M. guttatus and M. nasutus. Extensive sampling within a single M. guttatus population revealed that the hms1 locus is polymorphic and that the incompatibility allele appears to segregate at intermediate frequency, a pattern that is consistent with either genetic drift or natural selection.     

Postcopulatory selection for dissimilar gametes maintains heterozygosity in the endangered North Atlantic right whale

Although small populations are expected to lose genetic diversity through genetic drift and inbreeding, a number of mechanisms exist that could minimize this genetic decline. Examples include mate choice for unrelated mates and fertilization patterns biased toward genetically dissimilar gametes. Both processes have been widely documented, but the long‐term implications have received little attention. Here, we combined over 25 years of field data with high‐resolution genetic data to assess the long‐term impacts of biased fertilization patterns in the endangered North Atlantic right whale. Offspring have higher levels of microsatellite heterozygosity than expected from this gene pool (effect size = 0.326, P < 0.011). This pattern is not due to precopulatory mate choice for genetically dissimilar mates (P < 0.600), but instead results from postcopulatory selection for gametes that are genetically dissimilar (effect size = 0.37, P < 0.003). The long‐term implication is that heterozygosity has slowly increased in calves born throughout the study period, as opposed to the slight decline that was expected. Therefore, this mechanism represents a natural means through which small populations can mitigate the loss of genetic diversity over time.     

Homomorphic Self‐Incompatibility,with Stylar Rejection Site,in Luehea grandiflora Mart. and Zucc. (Tiliaceae‐Malvaceae s.l.)

Abstract: The breeding system of Luehea grandiflora (Tiliaceae‐Malvaceae s.l.) was investigated using hand pollinations and fluorescence microscopy studies of pollen tube growth. Although selfed flowers persisted for some 10 days, our study indicates that L. grandiflora is self‐incompatible, with self pollen tube inhibition in the upper style, as occurs in many taxa with homomorphic, gametophytic self‐incompatibility (GSI). L. grandiflora is only the second species reported within the Malvales with homomorphic stylar inhibition. This result is discussed within the context of a report for self‐compatibility in this species, and we also consider the phylogenetic implications for the occurrence of GSI in the family Malvaceae s.l.     

Spore killing in the fungus Podospora anserina: a connection between meiotic drive and vegetative incompatibility?

Fungi in which the haploid nuclei resulting from meiosis are linearly arranged in asci provide unique opportunities to analyse abnormal segregation. Any meiotic drive system in such fungi will be observed in a cross between a driving and a sensitive strain as spore killing: the degeneration of half the ascospores in a certain proportion of the asci. In a sample of some 100 strains isolated from a single natural population we have discovered at least six different meiotic drive elements (van der Gaag et al., 2000). Here we report results of research that was aimed at elucidating a possible correlation between meiotic drive and vegetative incompatibility in eight different Spore killer strains from this population. We show that there is a strong correlation between these two phenotypes, although the precise genetic nature of the correlation is not yet clear. We discuss the implications of our results for the understanding of the population genetics of meiotic drive in Podospora.