Nuclear and chloroplast DNA sequences data support the origin of Potamogeton intortusifolius J.B. He in China as a hybrid between P. perfoliatus Linn. and P. wrightii Morong

Internal transcribed spacers (ITS) of nuclear ribosomal DNA and chloroplast rbcL gene sequence data were used to test the hypothesis that natural populations of Potamogeton intortusifolius J.B. He in China originated from hybridization between P. perfoliatus Linn. and P. wrightii Morong. Based on ITS sequences data, P. intortusifolius possessed heterozygous rDNA genotypes which confirmed the hybrid origin of P. intortusifolius. Chloroplast rbcL gene sequences of P. intortusifolius from Yichang population revealed the same chloroplast haplotype as P. perfoliatus and the samples of P. intortusifolius from Weinan population had the same chloroplast haplotype as P. wrightii, which indicated that both putative parental species had been the maternal parent and that the two populations of P. intortusifolius had independent origins. This study confirms P. intortusifolius as a reciprocal hybrid. Because P. × intortusifolius in China has the same hybrid origin as P. × anguillanus Koidz. in Japan, it is suggested that P. × intortusifolius should be a synonym of P. × anguillanus.     

Using DNA‐based techniques to identify hybrids among linear‐leaved Potamogeton plants collected in China

Abstract It is well known that interspecific hybrids occur in the genus Potamogeton. The linear‐leaved Potamogeton species commonly have highly variable morphological characteristics. Their hybrids often show similar vegetative characters to their parental species and their identification based solely on morphology is not always conclusive. In order to clarify whether there are any hybrids from the linear‐leaved Potamogeton plants collected in China, we used internal transcribed spacers (ITS) of nuclear ribosomal DNA and chloroplast rbcL gene sequences to identify the hybrids. Using ITS sequence additivity, we identified four hybrids, namely P. orientalis (P. pusillus×P. oxyphyllus), P. pusillus×P. berchtoldii, P. foliosus×P. octandrus, and P. cristatus×P. octandrus. The latter three hybrids should be considered as new hybrids in Potamogeton. The maternal parents of the four hybrids were confirmed using chloroplast rbcL gene sequences.     

When interspecific prezygotic barriers break down: hybridization between two Potamogeton species (P. wrightii and P. perfoliatus) and comparison of the artificial and natural hybrids

Interspecific hybridization played an important role in speciation and evolution of angiosperms. Although the widespread occurrence of natural hybrids in the genus Potamogeton has been studied intensively, few successful experimental hybridization studies have been reported in this genus. In the present study, critical experimental hybridization was conducted using Potamogeton?×?intortusifolius, a natural hybrid widely distributed in China, and its parents (P. perfoliatus and P. wrightii). The absence of prezygotic barriers between P. wrightii and P. perfoliatus was observed, which contributes to the frequent hybridization of these species in nature. The pollen tube growth rates of P. perfoliatus were much faster than those of P. wrightii in the style of that species. However, the conspecific pollen tubes were competitively advantageous in P. perfoliatus styles. The interspecific hybridization could be applied bidirectionally, and 28 F1 hybrid individuals were successfully obtained from P. wrightii?×?P. perfoliatus, despite the low germination possibility of the hybrid seeds. Both the artificial and natural hybrids exhibited intermediate morphological characters but presented much lower fertility. The sterility of P.?×?intortusifolius is mainly due to its low female fitness. However, the offspring from P. wrightii?×?P.?×?intortusifolius indicated the potential for backcrossing in nature. This is the most successful attempt at artificial hybridization in this genus so far. The possible route for restoration of fertility and the fitness of the hybrids are also discussed in this paper.     

Phylogenetic relationships and hybrid origin of Potamogeton species (Potamogetonaceae) distributed in China: insights from the nuclear ribosomal internal transcribed spacer sequence (ITS)

Hybridization and polyploidization are important evolutionary processes in higher plants and have greatly enriched the diversity of the genus Potamogeton (Potamogetonaceae). To study the phylogenetic relationships and hybrid origin of Potamogeton species, 35 accessions representing 20 species, including diploids, tetraploids and hexaploids, and three hybrids were collected in China and their ribosomal internal transcribed spacers (ITS) were cloned, sequenced and statistically analyzed. The data showed that ITS sequences were informative to analyze the phylogeny of Potamogeton, and the phylogenetic tree revealed that Potamogeton species examined could be mainly divided into two groups (Group I and II), corresponding to subgenus Potamogeton and subgenus Coleogeton, respectively. Then, the evolutionary mechanism on the polyploidy of Potamogeton species was discussed. P. natans probably was an allotetraploid and one of its parent might result from aneuploidy change of species with 2n=28. P. hubeiensis might be derived from the hybridization between P. octandrus and P. cristatus. We suggested that both P. lucens and P. maackianus probably were allotetraploids, and P. obtusifolius might be a diploid hybrid between P. compressus and P. pusillus. Moreover, P. malainoides might have undergone biased concerted evolution toward one of its parent P. wrightii, and P. intortusifolius might be a synonymy of P. × anguillanus.     

Evidence for the hybrid origin of<Emphasis Type="Italic">potamogeton ×cooperi</Emphasis> (<Emphasis Type="Italic">Potamogetonaceae</Emphasis>): Traditional morphology-based taxonomy and molecular techniques in concert

The identity of plants morphologically intermediate betweenPotamogeton crispus andP. perfoliatus from two recently discovered sites, one in Moravia, Czech Republic and another in Wales, United Kingdom, was investigated with molecular markers. Evidence from restriction fragment length polymorphism analysis of the nuclear internal transcribed spacer region of ribosomal DNA and of thetrnK-trnQ chloroplast DNA intergenic spacer confirmed the morphology-based determination of two putative hybrid samples asP. ×cooperi. The hybrids showed the ITS variants of both parental taxa, consistent with the expected biparental inheritance of nuclear DNA. The chloroplast DNA markers indicateP. crispus as the female parent in both hybridization events. The hybrid origin of another dubious sample was excluded by the molecular data, in accordance with previous detailed morphological examination. This plant represented an extreme, narrow-leaved form ofP. perfoliatus, imitatingP. ×cooperi in some characters. The results of the molecular analyses are discussed in relation to the morphology of the plants. They underline that somePotamogeton hybrids could indeed be identified by careful and detailed morphological examination and also that these identifications were reliable and confirmed by molecular markers. This study exemplifies that long-term taxonomic expertise usually generates very well-founded specific questions suitable for straightforward treatment by appropriate molecular methods. The process and ecological implications of hybrid formation are also discussed     

Genetic diversity and origin of <Emphasis Type="Italic">Potamogeton anguillanus</Emphasis> (Potamogetonaceae) in Lake Biwa,Japan

We analyzed the genetic variation in Potamogeton anguillanus Koidz. and its putative parents, P. malaianus Miq. and P. perfoliatus L., at five allozyme loci of four enzymes to test the hypothesis of a hybrid origin for P. anguillanus, collected in Lake Biwa, Japan. Alleles diagnostic for either P. malaianus or P. perfoliatus were present at four loci. Of 13 single locus phenotypes (SLPs) of P. anguillanus, eight were phenotypes that were expected in F₁ hybrids between P. malaianus and P. perfoliatus. Two SLPs were different from those expected in F₁ hybrids but could have resulted from segregation of parental alleles in later generation hybrids. Each of the remaining three SLPs possessed one allele unique to P. anguillanus. Allozyme analyses thus supported the view that P. anguillanus was derived from hybridization between P. malaianus and P. perfoliatus. It seems likely that the genetic diversity of P. anguillanus found previously originated through multiple hybridizations and sexual processes in P. anguillanus. Other processes such as intragenic recombination, mutation, or hybridization with another lineage are also discussed with reference to the origin of unique alleles. Received: March 16, 2001 / Accepted: September 11, 2001     

Maternal effects and ecological divergence in aquatic plants: a case study in natural reciprocal hybrids between Potamogeton perfoliatus and P. wrightii

When parental taxa are adapted to different habitats, hybrid genotypes are often highly heterogeneous, such that habitat or ecological factors influence hybrid fate and ecological performance. Trait expression in hybrids is not always intermediate between the parents, but may instead be either parental‐like or extreme (transgressive) depending on genetic control of the phenotypes. Maternal effects arising from interspecific interaction between cytoplasmic and nuclear genomes are widely recognized as playing a role in character expression of natural hybrids. Such interaction often leads to hybrid sterility or inviability. When hybrids are viable, however, cytonuclear interaction may contribute to hybrid persistence through its influence on trait expression. To date, maternal influence on hybrid performance has been examined primarily in experimentally produced hybrids, or in natural hybrids without identification of the cross direction owing to difficulty in obtaining species‐specific molecular markers. In aquatic plants, many hybrids persist by extensive clonal growth and are important components of aquatic communities. Many such hybrids are known in Potamogeton (pondweeds), the largest aquatic genus. Because Potamogeton species are ecologically highly diverse and maternal lineages are readily distinguished using molecular markers, natural hybrids of Potamogeton are well‐suited for studies of maternal effects, especially those affecting vegetative performance. As a case study, we have focused on maternal effects on drought tolerance and depth distribution in the natural hybrid P. × anguillanus derived from the closely related species P. perfoliatus and P. wrightii.     

First record of Potamogeton×salicifolius for Italy,with isozyme evidence for plants collected in Italy and Sweden

Abstract

Potamogeton×salicifolius, a hybrid between P. lucens and P. perfoliatus, is identified as a new taxon for Italy. This is the first record of this hybrid from southern Europe. The Italian sample was studied in cultivation and compared with a living specimen of P.×salicifolius from Sweden, where the hybrid is rather widespread. In addition to morphological features, the most compelling evidence for the hybrid origin of these plants came from the isozyme analysis. The additive “hybrid” banding patterns of the five enzyme systems studied indicate inheritance from P. lucens and P. perfoliatus. The distribution of this hybrid in Europe coincides with the areas most severely affected by the Late Pleistocene glaciation. The relationships between environmental conditions, history of the habitat and rise of hybrids are discussed.     

Chloroplast evidence for the multiple origins of the hybrid Potamogeton × fluitans

Previous studies of Potamogeton × fluitans Roth. have supported the putative origin of this taxon as a result of hybridization between Potamogeton lucens L. and Potamogeton natans L. In the British Isles P. × fluitans is restricted to only a few sites, while both its parents are relatively common. Previous studies have shown that most of these hybrid populations are monomorphic, with the exception of the Moors River, in Hampshire and Dorset, where multiple genotypes have been identified. This is despite the fact that here the hybrid grows in the absence of one its parents, P. lucens. In this study, species-specific differences in the TrnL (UAA) chloroplast region, a maternally inherited marker, were used to identify the original seed-bearing parent of the hybrids within this river system. It was shown that both species have been maternal parents in the creation of this hybrid population.     

Inherited maternal effects on the drought tolerance of a natural hybrid aquatic plant, <Emphasis Type="Italic">Potamogeton anguillanus</Emphasis>

We tested whether maternal effects have led to the adaptive divergence of strains of the natural hybrid Potamogeton anguillanus, whose putative parents show contrastingly divergent ecologies. To examine the correlation between phenotypic characters and maternal types, we conducted drought experiments and DNA typing using nuclear and chloroplast genes. In the field, we investigated the distribution of the maternal type along the depth and the inshore-offshore gradient. Hybrids of P. malaianus mothers (M-hybrids) and those of P. perfoliatus mothers (P-hybrids) could not be distinguished morphologically under submerged conditions, but differed in drought tolerance. M-hybrids and P. malaianus formed more terrestrial shoots and exhibited higher survival than P-hybrids and P. perfoliatus in drought experiments. The distribution survey clarified that M-hybrids were dominant in shallow and inshore areas, whereas they were almost absent in deeper and offshore areas. These results indicate that the natural hybrid P. anguillanus differs in adaptive values depending on the maternal type. Bidirectional hybridization and heritable maternal effects may have played important roles in its phenotypic adaptation to local environmental conditions. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

A dynamic evolution of chromosome in subgenus Potamogeton revealed by physical mapping of rDNA loci detection

In this study, we analyzed 12 species in the subgenus Potamogeton (8 tetraploids and 2 diploids, 2 putative tetraploid hybrids) at the chromosomal level, including counting the chromosome number and physically mapping the rDNA. The extent of variation in the chromosome number and rDNA loci was determined in the Potamogeton species on both the inter- and intra-specific level. Moreover, one of the hybrid sets (P. perfoliatus ♂?×?P. wrightii ♀) was picked for performing artificial pollination for producing F1 generation adults and for sequential morphological character analysis and FISH detection. After comparing the parental species and natural hybrids from three different geographic locations, the intraspecific variations of rDNA loci were revealed in sampled plants; in addition, comparisons between artificial F1 and natural hybrids (P.?×?intortusifolius) showed a rapid change of 45S rDNA loci in response to the interspecific hybridization. We also compared our results concerning rDNA patterns with phylogenetic documents to derive complementary clues to karoytype evolution and interspecific relationships. Based on frequent hybridizations and active clonal reproduction with weak genetic selection, the rDNA chromosomal repatterning, such as the gain or loss of rDNA loci together with rDNA movements, might be one trend of chromosome evolution in this genus.     

Genetic Analysis of Putative Triploid Miscanthus Hybrids and Tetraploid M. sacchariflorus Collected from Sympatric Populations of Kushima, Japan

Miscanthus ?×giganteus, a triploid hybrid between tetraploid M. sacchariflorus and diploid M. sinensis, has considerable potential as a bioenergy crop. Currently only one genotype is widely cultivated, increasing its vulnerability to diseases during production. Finding new hybrids is important to broaden genetic resources of M. ×giganteus. Three putative triploid hybrids were discovered in a sympatric population of tetraploid M. sacchariflorus and diploid M. sinensis in Kushima, Japan. The hybrid nature of the triploids was determined by morphological analysis and sequencing the ribosomal DNA internal transcribed spacer (ITS) region. The triploids had awns on their florets, which is a common characteristic of diploid M. sinensis, and sheath hairs, which is typical of tetraploid M. sacchariflorus. All triploids showed heterozygosity in their ribosomal DNA ITS sequences. Based on these results, it is confirmed that the triploids are hybrids and novel genotypes of M. ×giganteus. Natural crossing between tetraploid M. sacchariflorus × diploid M. sinensis may also lead to the production of tetraploid hybrids. ITS analysis of tetraploid plants showed that one maternal parent of the triploid hybrids, K-Ogi-1, had heterozygous ITS, which was different than the other analyzed tetraploid, M. sacchariflorus. Thus, K-Ogi-1 was likely of hybrid origin. These tetraploid hybrids can also be utilized as parents in M. ×giganteus breeding. Since all hybrids identified in this study had tetraploid M. sacchariflorus as maternal parents, collecting and analyzing seeds from tetraploid M. sacchariflorus in sympatric areas could be an effective strategy to identify natural Miscanthus hybrids that can be used as bioenergy crops.     

Natural Hybridization and Introgression between Ligularia cymbulifera and L. tongolensis (Asteraceae,Senecioneae) in Four Different Locations

Natural hybridization has been considered to represent an important factor influencing the high diversity of the genus Ligularia Cass. in the Hengduan Mountains, China. Natural hybridization has been confirmed to occur frequently in Ligularia. To date, however, it has been demonstrated only within a single population. In this paper, we present evidence of natural hybridization in Ligularia from four different locations. The internal transcribed spacer (ITS) region of the nuclear ribosomal DNA and three chloroplast intergenic spacers (trnK-rps16, trnL-rpl32 and trnQ-5''rps16) of 149 accessions of putative hybrids and their putative parents (L. cymbulifera and L. tongolensis) were analyzed for evidence of hybridization. The ITS data clearly distinguished two putative parental species and sympatric L. vellerea and supported the hypothesis that those morphological intermediates were products of natural hybridization between L. cymbulifera and L. tongolensis. Moreover, several identified morphological parents were actual introgressed products. Because of hybridization and introgression, chloroplast DNA sequences generated a poorly resolved network. The present results indicate that varying degrees of hybridization and introgression occur differently depending on the habitat context. We conclude that gene flow caused by natural hybridization in Ligularia indeed plays an important role in the species diversity.     

Natural hybrids in Senecio (Asteraceae): new records from Argentina

Abstract

Hybridisation is very common within Senecio. It is considered a fundamental mechanism of speciation and also a major feature causing systematic complexity. Morphological features are the preliminary characteristics used to identify hybrids; however this approach is usually not sufficient when this is the case chromosome studies represent a valuable tool for reinforcing the observations. Two natural homoploid interspecific hybrids are reported here: Senecio × lulioi M. G. López et Xifreda (S. brasiliensis (Spreng.) Less. × S. grisebachii Baker) and Senecio × piepaloensis M. G. López et Xifreda (S. ragonesei Cabrera × S. sectilis Griseb.). Classic cytogenetic analysis and pollen stainability were performed to evaluate hybrid origins and to designate the putative parents. S. × lulioi presented a low frequency of quadrivalents and heteromorphic bivalents, both strong evidences of hybridisation between closely related species. S. × piepaloensis exhibited a high frequency of meiotic irregularities, and reduced pollen stainability, certain characteristics of hybrids. This analysis is discussed in relation to the role of hybridisation in the evolution of the genus, and its systematic significance.     

Interspecific hybridization in the genus Paeonia (Paeoniaceae): Polymorphic sites in transcribed spacers of the 45S rRNA genes as indicators of natural and artificial peony hybrids

The ITS1-5.8S rDNA-ITS2 regions of 33 accessions belonging to 16 species and five natural and garden interspecific hybrids of the genus Paeonia L. were sequenced. Chromatograms of the peony hybrids demonstrated the presence of the signals, corresponding to two different nucleotides at the positions differing in the parents, indicating that in the hybrids, no rDNA isogenization usually occurred, and they preserved rDNA of both parents. Analysis of these polymorphic sites (PS) showed that P. × majkoae was interspecific hybrid between P. tenuifolia and P. caucasica. The ITS of P. hybrida differs from ITS of P. × majkoae in 19 mutations. Because of this, P. × majkoae is definitely not synonymous to P. hybrida. Comparative analysis of ITS1-5.8S rDNA-ITS2 showed that species diversity in section Paeonia was based on recombination as a result of intraspecific hybridization of three haplotype families. Specifically, haplotypes A, typical of the P. tenuifolia and P. anomala genomes, haplotypes B, typical of P. mlokosewitschii and P. obovata, and haplotypes of family C, currently represented in rDNA of diploid and tetraploid forms of some Caucasian and Mediterranean species. The ITS regions many diploid peonies contain no dimorphic sites, while P. oreogeton, P. cambessedesii, P. rhodia, and P. daurica carry from 10 to 17 PS, and supposed to be the interspecific hybrids. Most of the tetraploid peonies contain from 6 to 18 PS in the ITS regions. These are alloploids with one of the parental genomes similar to that of P. mlokosewitschii (B1), or P. obovata (B3). The second parental genome in P. banatica, P. peregrina, and P. russii is represented by the genome, close to that of P. tenuifolia (A). P. macrophylla, P. mascula, P. coriacea, P. wittmanniana, and P. tomentosa carry genome of series B and genome of series C, which slightly resembles genome A.     

Maternal inheritance of chloroplast DNA in <Emphasis Type="Italic">Pinus mugo</Emphasis> Turra: a case study of <Emphasis Type="Italic">Pinus mugo</Emphasis> × <Emphasis Type="Italic">Pinus sylvestris</Emphasis> crossing

The opposite modes of chloroplast DNA (cpDNA) inheritance were found to operate in the reciprocal crossings of Scots pine (Pinus sylvestris L.) and mountain dwarf pine (Pinus mugo Turra). The crossings were found to be partially compatible. In P. sylvestris × P. mugo crossing, the paternal transmission of cpDNA to the offspring takes place corroborating the generally acknowledged concept of the paternal cpDNA inheritance in gymnosperms. On the contrary, in P. mugo × P. sylvestris crossing the seed progeny exhibited P. mugo haplotype of the mother tree deviating conspicuously from the above concept. In the open pollination offspring of the putatively hybrid individuals of the Scots and mountain dwarf pines, a biparental inheritance of cpDNA was revealed in mother tree with P. mugo haplotype indicating a loosened control of the maternal inheritance of cpDNA in the putative hybrids. Implications and impacts of this finding for further studies are discussed.     

Molecular characterization of two natural hybrids from the Iberian Peninsula: <Emphasis Type="Italic">Ranunculus × luizetii</Emphasis> and <Emphasis Type="Italic">R. × peredae</Emphasis> (Ranunculaceae)

Two nothospecies, Ranunculus × luizetii and R. × peredae (Ranunculaceae), were analyzed and discussed. For this purpose, Amplified Fragment Length Polymorphism (AFLP) markers, nuclear rDNA sequences (ITS1, 5.8S and ITS2) and pollen viability were conducted. The profiles of these hybrid samples were compared to their putative progenitors. Several additive polymorphic sites detected in the ITS sequences of the hybrid samples (R. × luizetii and R. × peredae) also confirmed their derived origins from ribotypes of their parental taxa (R. parnassiifolius subsp. parnassiifolius × R. pyrenaeus; R. amplexicaulis × R. cabrerensis subsp. cabrerensis, respectively). Despite the lack of exclusive AFLP markers reported in both hybrids, presumably due to effects of introgression, the concerted evolution of many rDNA polymorphisms towards either of the parental ribotypes indicated their ancient origin. Pollen fertility estimation in R. × luizetii presented a mean value of 60.58%, which showed that hybrid samples are well established and fertile. However, a larger difference was observed in R. × peredae, where the mean value of pollen fertility was very low (18.91%).     

Hybridization drives speciation in Gagea (Liliaceae)

Hybridization seems to play an important role in speciation of Gagea Salisb., a genus which is characterised by polyploid taxa lines and in which diploids (2n = 24) appear only to be common in basal sections. Hybrid detection was applied utilising direct and cloning nrDNA ITS data (ITS1, 5.8S rDNA, ITS2) combined with neighbour and ribotype networks and discussed in connection with previously published cpDNA, morphological and karyological data of the authors. We have evidence of the hybrid origin of taxa within the section Gagea (G. pomeranica, G. megapolitana) and the monophyletic clade of sections Didymobulbos and Fistulosae (G. microfistulosa, G. polidorii, G. cf. bohemica). Morphologically and karyologically differentiated Gagea megapolitana and G. pomeranica, adapted to synanthropic habitats, represent both hybrids of G. pratensis × G. lutea. Gagea microfistulosa represents a hybrid of G. villosa × G. fragifera; Gagea polidorii could represent the reverse hybrid. G. glacialis is also closely related to the latter complex.     

A redescription ofPotamogeton distinctus including remarks on the taxonomy of thePotamogeton nodosus group

Potamogeton distinctus has an E. Asian distribution and is closely related toP. nodosus. P. tepperi seems to be a synonym ofP. tricarinatus, even though the name has frequently been applied to other species likeP. distinctus. P. indicus is a synonym ofP. nodosus notwithstanding the fact that not all Indian broad-leaved pondweeds belong to that taxon.