Hybrid origins and F1 dominance in the free-floating, sterile bladderwort, Utricularia australis f. australis (Lentibulariaceae)

Abandonment of sexual reproduction is a well-known characteristic in aquatic plants, while the causes, levels, and consequences of sterility are often unknown. Utricularia australis f. australis (Lentibulariaceae) is a free-floating, sterile bladderwort distributed widely in temperate and tropical regions. Experimental crosses in cultivated conditions, AFLP analysis, and cpDNA haplotypes of natural populations clearly demonstrated that U. australis f. australis originates from the asymmetric hybridization between two parental taxa: U. australis f. tenuicaulis (mostly as female) and U. macrorhiza (mostly as male). No post-F(1) hybrids were detected using the additive patterns of AFLP bands combined with the observation of extensive sterility in U. australis f. australis. Recurrent hybridizations and subsequent perpetuation by asexual reproduction were demonstrated by the unique, but monomorphic, AFLP genotypes observed in each U. australis f. australis population. Hybrids and parental species did not coexist, implying the superiority of the hybrid U. australis f. australis in certain environmental conditions. It remains unclear whether populations of U. australis f. australis are maintained by colonizing propagules or as relicts of past hybridization events.     

Patterns, sources and ecological implications of clonal diversity in apomictic Ranunculus carpaticola (Ranunculus auricomus complex, Ranunculaceae)

Sources and implications of genetic diversity in agamic complexes are still under debate. Population studies (amplified fragment length polymorphisms, microsatellites) and karyological methods (Feulgen DNA image densitometry and flow cytometry) were employed for characterization of genetic diversity and ploidy levels of 10 populations of Ranunculus carpaticola in central Slovakia. Whereas two diploid populations showed high levels of genetic diversity, as expected for sexual reproduction, eight populations are hexaploid and harbour lower degrees of genotypic variation, but maintain high levels of heterozygosity at many loci, as is typical for apomicts. Polyploid populations consist either of a single AFLP genotype or of one dominant and a few deviating genotypes. genotype/genodive and character incompatibility analyses suggest that genotypic variation within apomictic populations is caused by mutations, but in one population probably also by recombination. This local facultative sexuality may have a great impact on regional genotypic diversity. Two microsatellite loci discriminated genotypes separated by the accumulation of few mutations ('clone mates') within each AFLP clone. Genetic diversity is partitioned mainly among apomictic populations and is not geographically structured, which may be due to facultative sexuality and/or multiple colonizations of sites by different clones. Habitat differentiation and a tendency to inhabit artificial meadows is more pronounced in apomictic than in sexual populations. We hypothesize that maintenance of genetic diversity and superior colonizing abilities of apomicts in temporally and spatially heterogeneous environments are important for their distributional success.     

Photosynthetic CO2 affinity of the aquatic carnivorous plant Utricularia australis (Lentibulariaceae) and its investment in carnivory

Aquatic carnivorous plants usually grow in shallow dystrophic waters poor in inorganic N and P. Utricularia australis was chosen as a model plant for its prolific distribution and great ecological plasticity. The photosynthetic CO₂ compensation point and factors associated with investment in carnivory and capture of prey were measured in 17 U. australis micropopulations in Třeboň basin, Czech Republic, together with water chemistry factors at these sites differing greatly in their trophic level, water hardness, and prey availability. Apical shoot growth rate was estimated at some oligotrophic sites. The micropopulations differed greatly in the proportion of traps with animal prey (2.7–70%, mean 26%), trap proportion to total biomass (1.4–42%, mean 26%), mean trap biomass (0.7–63 μg trap⁻¹, mean 19 μg), and maximum trap size (1–3 mm, mean 2.0 mm). CO₂ compensation points ranged from 0.7 to 6.1 μM (mean 2.6 μM). A weak HCO₃ ⁻ use (compensation point 0.51 mM) was found in plants growing in alkaline water. Trap biomass proportion did not correlate significantly with prey capture and CO₂ compensation points with ambient [CO₂]. A very rapid apical growth (2.5–4.2 new nodes day⁻¹) occurred in sand pits. Thus, HCO₃ ⁻ use in U. australis can be induced by growing at very high pH. CO₂ compensation points resembled those known in other aquatic non-carnivorous plants. They did not reflect carnivory. In spite of very rapid apical shoot growth, the relative growth rate of U. australis can be zero in oligotrophic habitats without prey.     

Genetic structure in aquatic bladderworts: clonal propagation and hybrid perpetuation

• Background and Aims The free-floating aquatic bladderwort Utricularia australis f. australis is a sterile F₁ hybrid of U. australis f. tenuicaulis and U. macrorhiza. However, co-existence of the hybrids and parental species has not been observed. In the present study, the following questions are addressed. (a) Does the capacity of the two parental species to reproduce sexually contribute to higher genotypic diversity than that of sterile F₁ hybrid? (b) Are there any populations where two parental species and their hybrid co-exist? (c) If not, where and how do hybrids originate?• Methods The presence and absence of Utricularia was thoroughly investigated in two regions in Japan. An amplified fragment length polymorphism (AFLP) analysis was conducted for 397 individuals collected from all populations (33 in total) where Utricularia was observed.• Key Results The mean number of genotypes per population (G) and genotypic diversity (D) were extremely low irrespective of the capacity to reproduce sexually: G was 1·1–1·2 and D was 0·02–0·04. The hybrid rarely co-existed with either parental species, and the co-existence of two parental species was not observed. Several AFLP bands observed in the hybrid are absent in both parental genotypes, and parent and hybrid genotypes in the same region do not show greater genetic similarity than those in distant regions.• Conclusions The capacity to reproduce sexually in parental species plays no role in increasing genotypic diversity within populations. The observed genotypes of the hybrid could not have originated from hybridization between the extant parental genotypes within the study regions. Considering the distribution ranges of three investigated taxa, it is clear that the hybrid originated in the past, and hybrid populations have been maintained exclusively by clonal propagation, which may be ensured by both hybrid vigor and long-distance dispersal of clonal offspring.     

Sexual reproduction, clonal diversity and genetic differentiation in patchily distributed populations of the temperate forest herb Paris quadrifolia (Trilliaceae)

Clonal plant species have been shown to adopt different strategies to persist in heterogeneous environments by changing relative investments in sexual reproduction and clonal propagation. As a result, clonal diversity and genetic variation may be different along environmental gradients. We examined the regional and local population structure of the clonal rhizomatous forest herb Paris quadrifolia in a complex of forest fragments in Voeren (Belgium). Relationships between population size (the number of shoots), shoot density (the number of shoots per m²) and local growth conditions were investigated for 47 populations. Clonal diversity and genetic variation within and among 19 populations were investigated using amplified fragment length polymorphism markers. To assess the importance of sexual reproduction, seed set, seed weight and germination success were determined in 18 populations. As predicted, local growth conditions largely affected population distribution, size and density of P. quadrifolia. Populations occurring in moist and relatively productive sites contained significantly more shoots. Here, shoots were also much more sparsely distributed compared to populations occurring in dry and relatively unproductive sites, where shoots showed a strongly aggregated distribution pattern. Clonal diversity was relatively high, compared with other clonal species (G/N ratio = 0.43 and Simpson’s D=0.81). Clonal diversity significantly (P<0.01) decreased with increasing shoot density while molecular genetic variation was significantly (P<0.01) affected by population size and local environmental conditions. Lack of recruitment and out-competition of less-adapted genotypes may explain the decreased genetic variation in dry sites. Analysis of molecular variance revealed significant genetic variation among populations (Φ _ST=0.42, P<0.001), whereas pairwise genetic distances were not correlated to geographic distances, suggesting that gene flow among populations is limited. Finally, the number of generative shoots, the number of seeds per fruit and seed weight were significantly and positively related to population size and local growth conditions. We conclude that under stressful conditions populations of clonal forest plant species can slowly evolve into remnant populations characterized by low levels of genetic variation and limited sexual reproduction. Conservation of suitable habitat conditions is therefore a prerequisite for effective long-term conservation of clonal forest plant species.     

Sexual and vegetative reproduction in the aboveground part of a dioecious clonal plant, <Emphasis Type="Italic">Dioscorea japonica</Emphasis> (Dioscoreaceae)

In dioecious clonal plants, the reproductive effort required to set seeds will be responsible for the larger investment in sexual reproduction by females. If there will be a trade-off in resource allocation between sexual and clonal reproduction, this differential sexual reproduction will lead to sexual differentiation in the relative amount of clonal reproduction. To test this prediction, we studied differences between the sexes in their phenologies and investments in sexual and vegetative reproduction (clonal reproduction by means of bulbils) with respect to ramet size in a dioecious clonal plant, Dioscorea japonica Thunb. The period of bulbil production overlapped the period during which infructescences developed. Females flowered later, produced heavier inflorescences, and fewer flowers per inflorescence than did males. Regression analysis using the size of the individual plants demonstrated that large females made smaller investments in inflorescences and larger investments in sexual reproduction than did large males. In contrast, females invested fewer resources in vegetative reproduction than did males. However, the total investments in sexual and vegetative reproduction did not differ between the sexes. These results supported our hypothesis on the sexual differentiation in sexual and clonal reproduction.     

Causes and consequences of extreme variation in reproductive strategy and vegetative growth among invasive populations of a clonal aquatic plant, <Emphasis Type="Italic">Butomus umbellatus</Emphasis> L. (Butomaceae)

The mode of reproduction may influence the spread of invasive species by affecting evolutionary potential and dispersal ability. We sampled 51 introduced North American populations of the clonal aquatic plant Butomus umbellatus L. (flowering rush) and found extreme variation in sexual fertility caused by polyploidy. Populations consisted of diploids that produced thousands of viable seeds or of sexually sterile triploids. Although a trade-off between sexual and clonal reproduction predicts that the sexual sterility of triploids would be compensated for by greater clonal reproduction, a greenhouse experiment involving eight diploid and 10 triploid populations showed that diploid plants not only invest substantially in sexual structures but also make hundreds of tiny clonal bulbils on both rhizomes and inflorescences. In contrast, triploids do not make bulbils and have very limited scope for clonal multiplication and dispersal. Diploid populations were more frequent than triploid populations, especially in the Great Lakes region. This is probably because of the difference between cytotypes in clonal rather than sexual reproduction, as genetic analyses indicate a general lack of sexual recruitment in North America. Although triploids were less common, they have a wider geographical distribution. This could be due to a greater ecological tolerance resulting from polyploidy. However, genetic evidence suggests that triploids have become widespread via their use in and escape from horticulture. North America is being colonized by two distinct forms of B. umbellatus that differ strongly in reproductive strategy as well as the vectors and pathways of invasion.     

Clonal structure and patterns of allozyme diversity in the rare endemic Cycladenia humilis var. jonesii (Apocynaceae)

The rare endemic Cycladenia humilis var. jonesii (Jones cycladenia) has low levels of sexual reproduction. Enzyme electrophoresis was used to explore possible causes of low seed set and high fruit abortion by assessing the clonal structure and genetic diversity in populations. The seven populations studied were composed of multiple, highly interdigitated clones; thus low fruit set is not likely to be due to a scarcity of mates. Genotype frequencies did not differ significantly from Hardy-Weinberg proportions, suggesting that populations are not highly inbred. Jones cycladenia exhibited high levels of genetic diversity at both the population level (A = 1.7; P = 37; He = 0.14) and the taxon level (A = 2.7; P = 60) in comparison to other plants. These data suggest that genetic drift is unlikely to have left this taxon genetically depauperate. Furthermore, we detected little divergence among geographically disjunct populations of Jones cycladenia (FST = 0.10). In comparison, Jones cycladenia populations were highly differentiated from a population of the taxon's close relative, C. h. var. humilis (mean genetic identity = 0.76). Our study suggests that other reasons for low fruit set in Jones cycladenia, such as resource or pollinator limitation, or genetic load, should be explored in future research.     

Long-distance seed dispersal, clone longevity and lack of phylogeographical structure in the European distributional range of the coastal Calystegia soldanella (L.) R. Br. (Convolvulaceae)

Aim  To explore the relative effects of Quaternary climatic history vs. species-specific biological properties (high seed dispersability, high seed longevity, clonal growth) on phylogeographical structure in European Mediterranean and Atlantic Ocean and Black Sea material of the coastal dune plant Calystegia soldanella (L.) R. Br.
Location  Black Sea and European Mediterranean and Atlantic Ocean coasts.
Methods  Variation in amplified fragment length polymorphism was analysed at two different sampling levels. First, an entire-range sample from the Black Sea to the North Sea, including single individuals from sites evenly spread along this entire coast was analysed. Second, in a population-level sample, seven populations from the European Mediterranean and Atlantic Ocean coasts were analysed.
Results  Neither the entire-range nor the population-level sampling resulted in clear phylogeographical patterns. Instead, individuals from geographically distant areas were often genetically more similar to each other than individuals from the same area. Non-significant isolation-by-distance was found for both sampling approaches, and comparatively low levels of intrapopulational genetic variation were observed.
Main conclusions  The lack of phylogeographical structure in C. soldanella , in comparison with the clear phylogeographical patterns observed in other coastal plant species analysed previously, is postulated to be the result of the specific biology of C. soldanella . The combination of high seed longevity, high dispersability of seeds by sea water and clonal growth and probable high clone age are likely to be responsible for the observed absence of phylogeographical structure. This implies that extreme biological properties such as those shown by C. soldanella can either erase or prevent the formation of historical patterns of genetic variation.     

Genetic impoverishment and cross-incompatibility in remnant genotypes of Ziziphus celata (Rhamnaceae), a rare shrub endemic to the Lake Wales Ridge, Florida

The loss of genetic diversity in fragmented populations ofself-incompatible plant species may result in sexual reproductive failure andlocal extinctions. Florida ziziphus (Ziziphus celata) is aself-incompatible clonal shrub known only from five genetically depauperatepopulations on the Lake Wales Ridge, Florida, USA. Recovery of this speciesrequires identification of cross-compatible genotypes that can be used to createviable (i.e., sexually reproducing) populations. To further development of arecovery program for this highly imperiled species, we investigated its geneticstructure and sexual reproductive viability. We used random amplifiedpolymorphic DNAs (RAPDs) to investigate genetic variability within remnantpopulations and we conducted experimental compatibility trials to determine thecross-compatibility of remnant genotypes. One hundred and ninety-nine uniquestem samples collected from one ex situ and fivein situ populations were assayed for the presence orabsence of a band for 32 RAPD markers. Based on unweighted pair-group meancluster analysis (UPGMA), only 11 multi-locus genotypes (MLGs) were identified.Eight of these MLGs correspond to MLGs identified in an earlier allozyme study.In addition, we identified three new RAPD-based MLGs. Three of the five naturalpopulations consisted of only one MLG, while the largest and most geneticallydiverse population comprised only four MLGs. Coefficients of similarity rangedfrom 96.6% for the most closely related MLGs to 20.7% for the most distantlyrelated. The compatibility trials demonstrated that most MLGs arecross-incompatible. With 69% of all possible one-way crosses tested (38/55), wehave identified only eight compatible crosses via germination trials. Based onthe results of the compatibility trials, we assigned MLGs toself-incompatibility (SI) mating types. On present evidence, the currentbreeding population of Florida ziziphus may comprise as few as two SI matingtypes. These SI mating types will be used to guide future breeding efforts andan experimental introduction.     

Introduction of SLG (S locus glycoprotein) alters the phenotype of endogenous S haplotype,but confers no new S haplotype specificity in Brassica rapa L.

Self-incompatibility (SI) in Brassicaceae is genetically controlled by the S locus complex in which S locus glycoprotein (SLG) and S receptor kinase (SRK) genes have been identified, and these two genes encoding stigma proteins are believed to play important roles in SI recognition reaction. Here we introduced the SLG⁴³ gene of Brassica rapa into a self-incompatible cultivar, Osome, of B. rapa, and examined the effect of this transgene on the SI behavior of the transgenic plants. Preliminary pollination experiments demonstrated that Osome carried S⁵² and S⁶⁰, and both were codominant in stigma, but S⁵² was dominant to S⁶⁰ in pollen. S⁴³ was found to be recessive to S⁵² and codominant with S⁶⁰ in stigma. The nucleotide sequence of SLG⁴³ was more similar to that of SLG⁵² (87.8% identity) than to that of SLG⁶⁰ (74.8% identity). Three of the ten primary transformants (designated No. 1 to No. 10) were either completely (No. 9) or partially (No. 6 and No. 7) self-compatible; the SI phenotype of the stigma was changed from S⁵²S⁶⁰ to S⁶⁰, but the SI phenotype of the pollen was not altered. In these three plants, the mRNA and protein levels of both SLG⁴³ and SLG⁵² were reduced, whereas those of SLG⁶⁰ were not. All the plants in the selfed progeny of No. 9 and No. 6 regained SI and they produced a normal level of SLG⁵². These results suggest that the alteration of the SI phenotype of the stigma in the transformants Nos. 6, 7, and 9 was the result of specific co-suppression between the SLG⁴³ transgene and the endogenous SLG⁵² gene. Three of the transformants (Nos. 5, 8 and 10) produced SLG⁴³ protein, but their SI phenotype was not altered. The S⁶⁰ homozygotes in the selfed progeny of No. 10 which produced the highest level of SLG⁴³ were studied because S⁴³ was codominant with S⁶⁰ in the stigma. They produced SLG⁴³ at approximately the same level as did S⁴³S⁶⁰ heterozygotes, but did not show S⁴³ haplotype specificity at the stigma side. We conclude that SLG is necessary for the expression of the S haplotype specificity in the stigma but the introduction of SLG alone is not sufficient for conferring a novel S haplotype specificity to the stigma.     

Host plant interactions of the corn planthopper, Peregrinus maidis Ashm. (Homoptera: Delphacidae) in maize and sorghum agroecosystems

The corn planthopper, Peregrinus maidis (Ashmead) (Homoptera: Delphacidae) causes serious economic losses in corn and sorghum. The insect occurs mostly at humid low elevations in the tropics and coastal areas of subtropical and temperate regions of all continents, the Caribbean Islands, and islands in the Atlantic, Indian, and Pacific Oceans. This review provides a detailed compilation on the chronological progress made in basic and strategic aspects of research on the interactions between P. maidis and various host plants. The nature of damage by P. maidis and its economic impact, ecobiology in relation to host diversity, abiotic, and seasonal interactions; and life tables and alary polymorphism are discussed. Host plant resistance studies indicate that very few sources of resistance to P. maidis have been identified in maize, sorghum, or pearl millet, warranting a need to standardize rapid and reliable screening methods. The behavioral responses vis-à-vis mechanisms of resistance show the predominance of antixenosis for colonization and/or oviposition with variable degrees of antibiosis affecting life cycle parameters of P. maidis on maize and sorghum. The role of morphological traits, physiological mechanisms, and biochemical factors governing resistance are described. Population dynamics based on density-dependent and density-independent interactions are also discussed. In addition, aspects of P. maidis on chemical control, biological control, and trophobiosis interactions are listed. Future thrusts on research approaches are also discussed. Genetic engineering techniques involving lectin genes in the development of transgenic plants, and the molecular mapping of genes conferring resistance to both P. maidis and its transmitted virus diseases may stimulate further research and lead to better understanding of P. maidis—host plant interactions. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.