Hybridisation in the genera Vulpia and Festuca: the production of artificial F1 plants

The results of hybridisation experiments involving 24 strains of diploid, tetraploid and hexaploid Vulpia , representing nine species in three sections, and 22 strains of hexaploid and octoploid Festuca rubra agg., are presented, and the taxonomic implications discussed. 7848 pollinations produced 741 hybrid caryopses, from which 137 mature hybrid plants were raised (1.75% success rate) by embryo culture. Festuca x Festuca crosses were relatively easily achieved, and Festuca x Vulpia crosses were as successful as Vulpia x Vulpia crosses, indicating a close relationship between these two genera. Hybrids with F. rubra agg. were raised from females of species from all three sections of Vulpia. Within Vulpia , the most successful crosses involved V. geniculata, V. ligustica, V. fasciculata and V. pyramidata , whereas V. alopecuros was the least successful parent of all, producing only one mature hybrid from 2595 pollinations.     

Hybridization in the genera Vulpia and Festuca (Poaceae): meiotic behaviour of artificial hybrids

The course of meiosis, including an analysis of chromosome configurations, is described for five diploid × diploid Vulpia crosses, five tetraploid × diploid Vulpia crosses, one hexaploid × diploid Festuca × Vulpia cross, one tetraploid × hexaploid Vulpia × Festuca cross, and one hexaploid × hexaploid Vulpia × Festuca cross. In most cases there was 97.5% or more pollen sterility, but two heptaploid plants obtained (presumably by non-reduction) from a hexaploid × diploid cross had about 60% stainable pollen. In the diploid hybrids pairing was quite extensive, and in V. ligustica × V. geniculata it was more or less as in the parent species (mode 7 bivalents, with regular separation). In the triploid hybrids the modal situation was 7 bivalents + 7 univalents, but evidence concerning the genomes which were pairing was equivocal. Evidence from the crosses at higher ploidy levels shows that both homogenetic and heterogenetic pairing does occur, although the relative amounts are uncertain. The results in general support the current classsification of Vulpia , except that they suggest the removal of V. alopecuros from section Loretia.     

Continuing addition to the gene-pool of theFestuca rubra aggregate (Poaceae:Poeae)

The chance backcrossing to hexaploidFestuca rubra of a natural pentaploid hybridFestuca rubra ·Vulpia fasciculata to produce a hexaploid plant is described. The hexaploid derivative, unlike the pentaploid hybrid, shows a reasonable degree of fertility and is morphologically very close to normalFestuca rubra. Its existence provides a mechanism for the continued introgression ofVulpia fasciculata intoFestuca rubra, so increasing the variability of the latter. It also shows that pairing betweenFestuca rubra andVulpia fasciculata chromosomes must have taken place either in the pentaploid hybrid or in its hexaploid derivative. A possibly similar pattern of introgression involvingVulpia bromoides is also described.     

Variation in the Festuca brachyphylla (Poaceae) complex in Svalbard, elucidated by chromosome numbers and isozymes

Contrasting with former taxonomic treatments, chromosome numbers and isozyme data support the delimitation of the seminiferous representatives of the Festuca brachyphylla complex in Svalbard into four species: F. baffinensis, F. brachyphylla, F. hyperborea and F. edlundiae. Unique enzyme markers were found for all species. Festuca brachyphylla proved hexaploid, and the others, tetraploid. The chromosome numbers of F. hyperborea and F brachyphylla (as circumscribed at present) are new to Svalbard. Festuca baffinensis is the most distinct species within the complex, probably representing a separate evolutionary lineage. The three other species seem closely related, showing mutually equidistant relationships. Some deviating plants found on disturbed ground might represent hybrid derivatives or an introduced foreign strain of the elsewhere variable F. brachyphylla. Materials of diploid F. ovina from northern Fennoscandia was enzymatically closely related to the F brachyphylla complex in Svalbard. Festuca brachyphylla, F. edlundiae , and F. hyperborea all had a stronger affinity to F ovina than to F baffinensis , indicating that the F brachyphylla complex is an artificial taxonomic group. There are reasons to believe that the origin of the polyploid taxa of the F brachyphylla complex can be traced to diploid species of the F. brachyphylla and F ovina complexes.     

云南宾川县鸡足山地区多星韭种群分化的研究

对鸡导山地区７个多星韭种群的生物生态学特性进行了比较全面的研究。结果表明;鸡足山地区的多星韭有二倍体和四倍体２种倍性。多星韭二倍体比四倍体更能忍受较恶劣的环境条件。二倍体多星韭生境单一,种群分化不明显;四倍体多星韭生境多样,种群间变异丰富,分化强烈。     

Estimation of preferential pairing in tetraploid x diploid hybridizations

Summary Crosses made between tetraploid and diploid, 2n pollen-producing species directly transfer from one-half to the entire diploid genome from the diploid to the tetraploid level, depending on the mechanism of 2n pollen formation and the amount of crossing-over that occurs. Tetraploid plants that result from tetraploid x diploid hybridizations can be further utilized in a breeding program. It is postulated that preferential pairing between homologous chromosomes derived from the original tetraploid or diploid parent occurs in the tetraploid x diploid hybrid. Depending on the genetic divergence of the species involved, preferential pairing of homologous chromosomes may range from zero to one. Theoretical estimates of the amount of preferential pairing and the standard errors of these estimates are derived for cases where the diploid parent produces 2n gametes by either a first division or a second division restitution mechanism.     

Spread of violets in polluted pine forests: morphological and molecular evidence for the ecological importance of interspecific hybridization

Hybridization between plant species occurs frequently but hybrids are often restricted to ecotones or disturbed habitats. In this study we show that introgressive hybrids between the tetraploid Viola riviniana and the diploid V. reichenbachiana invaded pine forests of the Dübener Heide (central Germany), an area affected by calcareous pollutants. The spread of these violet populations was correlated with the impact of pollution on habitat conditions. We compared morphology, cytology and random amplified polymorphic DNA (RAPD) bands among six Viola populations from the Dübener Heide and three populations of each pure species. RAPD analysis using 12 primers revealed 141 scorable bands. We considered bands as species specific if they occurred in at least 75% of the plants in one pure violet species but in none of the other. Seven bands were specific to V. riviniana and 11 bands were specific to V. reichenbachiana . Two plants of a V. reichenbachiana population were identified as hybrids. Of the Viola populations from the Dübener Heide, one was diploid and could be classified as V. reichenbachiana by morphology and RAPD markers. However, the majority of the Dübener Heide populations were tetraploid, and of a more variable morphology than V. riviniana and V. reichenbachiana , showing different combinations of intermediate characters, characters of the pure species and extreme characters. Despite their overall genetic similarity to V. riviniana , these plants could be identified as introgressive hybrids between V. riviniana and V. reichenbachiana by species-specific RAPD bands. Therefore, we propose that recurrent hybridization and backcrossing resulted in novel genotypes adapted to the changed environment of polluted pine forests.     

Cytological,morphological and molecular analyses of controlled progenies from meiotic mutants of alfalfa producing unreduced gametes

A program of sexual polyploidization was carried out in alfalfa using plants from wild diploid species that produced male or female unreduced gametes. Sixteen progenies from 2x-4x and 2x-2x crosses were examined with a combination of morphological, cytological and molecular analyses. The chromosome counts revealed diploid, tetraploid and aneuploid plants. Plants with B chromosomes were also detected. The leaf area of the plants was a useful characteristic for distinguishing tetraploid from diploid plants obtained by unilateral or bilateral sexual polyploidization. Leaf shape and leaf margin were not correlated with the ploidy levels. Plants with supernumerary chromosomes displayed obovate or elliptic leaves which differed markedly from the range of forms typical of diploid and tetraploid alfalfa plants. RAPD markers were investigated in all progeny plants to determine maternal and paternal amplification products. Three alfalfa-specific primers proved to be effective in revealing the hybrid origin of the plants. A combination of cytological, morphological and molecular analyses is essential for a detailed genetic characterization of progenies in programs of sexual polyploidization.     

Spread of violets in polluted pine forests: morphological and molecular evidence for the ecological importance of interspecific hybridization

Hybridization between plant species occurs frequently but hybrids are often restricted to ecotones or disturbed habitats. In this study we show that introgressive hybrids between the tetraploid Viola riviniana and the diploid V. reichenbachiana invaded pine forests of the Dübener Heide (central Germany), an area affected by calcareous pollutants. The spread of these violet populations was correlated with the impact of pollution on habitat conditions. We compared morphology, cytology and random amplified polymorphic DNA (RAPD) bands among six Viola populations from the Dübener Heide and three populations of each pure species. RAPD analysis using 12 primers revealed 141 scorable bands. We considered bands as species specific if they occurred in at least 75% of the plants in one pure violet species but in none of the other. Seven bands were specific to V. riviniana and 11 bands were specific to V. reichenbachiana . Two plants of a V. reichenbachiana population were identified as hybrids. Of the Viola populations from the Dübener Heide, one was diploid and could be classified as V. reichenbachiana by morphology and RAPD markers. However, the majority of the Dübener Heide populations were tetraploid, and of a more variable morphology than V. riviniana and V. reichenbachiana , showing different combinations of intermediate characters, characters of the pure species and extreme characters. Despite their overall genetic similarity to V. riviniana , these plants could be identified as introgressive hybrids between V. riviniana and V. reichenbachiana by species-specific RAPD bands. Therefore, we propose that recurrent hybridization and backcrossing resulted in novel genotypes adapted to the changed environment of polluted pine forests.     

Evolution in an autopolyploid group displaying predominantly bivalent pairing at meiosis: genomic similarity of diploid Vaccinium darrowi and autotetraploid V. corymbosum (Ericaceae)

The genomic relationship between V. darrowi Camp (2n = 2x = 24) and V. corymbosum L. (2n = 4x = 48) was examined using an interspecific tetraploid hybrid, US 75, and representatives of the parental species. Two features in the background of US 75 led to the prediction that it was an allopolyploid: (1) the parental species are quite distinct morphologically and geographically, and (2) the diploid genome was incorporated into US 75 via an unreduced gamete. However, US 75 recently was shown to display tetrasomic inheritance using molecular markers. In the present cytological study, US 75 was found to have a lower than expected number of multivalents for an autopolyploid, although it had a significantly higher number of quadrivalents than its autotetraploid parent, V. corymbosum. Normal chromosome distributions were observed at anaphase I and II, and pollen viability was high. Our findings suggest that little genomic divergence has developed between the Vaccinium species and that the polyploids may freely exchange genes with sympatric diploid species via unreduced gametes. This pattern of hybridization could be an important component of evolution in all autopolyploid groups, making them much more dynamic than traditionally assumed.     

THE PROBLEM OF DRYOPTERIS LEEDSII

Walker , S. (U. Liverpool, England.) The problem of Dryopteris leedsii. Amer. Jour. Bot. 49(9): 971–974. Illus. 1962.—Investigation of 12 plants of ‘Dryopteris leedsii‘ from the type locality in Harford County, Maryland, has revealed 3 cytological entities, diploid, triploid and tetraploid. The diploid and triploid types exhibit hybrid characters and are sterile, while the tetraploid is a fertile species. The relationship between the 3 entities remains a problem, but the possibilities are discussed.     

Genome relationships in the genus Dasypyrum (Gramineae)

To elucidate the genome relationships in the genus Dasypyrum and the ancestry of tetraploid D. breviaristatum, two cytotypes of D. breviaristatum and D. villosum were reciprocally crossed with one another. Chromosome pairing at the first metaphase of meiosis and fertility were examined in the F1 hybrids and the parental plants. The mean pairing configuration and mean arm pairing frequency in D. villosum-D. breviaristatum (2x) hybrids were 11.12I + 1.44II per cell and 0.107, respectively, and they were almost completely sterile. In D. breviaristatum (4x)-D. breviaristatum (2x) hybrid, up to seven trivalents were formed, and the mean pairing configuration was 3.38I + 3.20II + 3.74III + 0.005IV per cell. The mean arm pairing frequency and relative affinity calculated in that F1 hybrid were 0.915 and 0.641, respectively. Seven bivalents and seven univalents were characteristically formed in D. villosum-D. breviaristatum (4x) hybrids. Based on the present results, we clearly concluded that the genome of diploid D. breviaristatum is distantly related to the genome V of D. villosum, and that these two species have different basic genomes. We, therefore, proposed the symbol Vb for the haploid genome of diploid cytotype of D. breviaristatum. Moreover, we concluded that tetraploid D. breviaristatum is an autotetraploid with doubled sets of the genomes homologous with that of diploid D. breviaristatum, and we proposed the genome constitution VbVb for the haploid genome set of tetraploid cytotype of D. breviaristatum. Furthermore, from the chromosome pairing in the F1 hybrids involving Moroccan and Greek accessions, it was suggested that complicated rearrangements of chromosome structure have occurred in tetraploid D. breviaristatum in its natural populations across the entire distribution area.     

秋水仙碱诱导重瓣大岩桐(Sinningia speciosa)多倍体的研究

以重瓣大岩桐叶片为外植体,经秋水仙碱处理得到大量的多倍体植株。在培养基中加入秋水仙碱２０ｍｇ Ｌ＾－１处理一周,可使重瓣大岩桐的诱变率达到６２．５％,对再生植株进行形态学观察表明,多倍体植株比二倍体的茎粗壮,叶片增大,加厚。细胞学鉴定四倍体染色体数为２ｎ＝４ｘ＝５２,而二倍体的染色体数为２ｎ＝２６。     

Evidence for non-disomic inheritance in a Citrus interspecific tetraploid somatic hybrid between C. reticulata and C. limon using SSR markers and cytogenetic analysis

Artificial tetraploid somatic hybrids have been developed for sterile triploid citrus breeding by sexual hybridization between diploid and tetraploid somatic hybrids. The genetic structure of diploid gametes produced by tetraploid genotypes depends on the mode of chromosome association at meiosis. In order to evaluate tetraploid inheritance in a tetraploid interspecific somatic hybrid between mandarin and lemon, we performed segregation studies using cytogenetic and single sequence repeat molecular markers. Cytogenetic analysis of meiosis in the somatic hybrid revealed 11% tetravalents and 76% bivalents. Inheritance of the tetraploid hybrid was analyzed by genotyping the triploid progeny derived from a cross between a diploid pummelo and the tetraploid somatic hybrid, in order to derive genotypes of the meiospores produced by the tetraploid. A likelihood-based approach was used to distinguish between disomic, tetrasomic, and intermediate inheritance models and to estimate the double reduction rate. In agreement with expectations based the cytogenetic data, marker segregation was largely compatible with tetrasomic and inheritance intermediate between disomic and tetrasomic, with some evidence for preferential pairing of homoeologous chromosomes. This has important implications for the design of breeding programs that involve tetraploid hybrids, and underscores the need to consider inheritance models that are intermediate between disomic and tetrasomic.     

Absence of the J genome in Leymus species (Poaceae: Triticeae): evidence from DNA hybridization and meiotic pairing.

To test the presence of a J genome in the type species of Leymus, L. arenarius, its total genomic DNA and that of tetraploids L. mollis, L. salinus ssp. salmonis, L. ambiguus, L. chinensis, L. secalinus, L. alaicus ssp. karataviensis, and L. innovatus were probed with the 277-bp insert of pLeUCD2, which can hybridize with the J, S, and P but not with the N, R, V, Q, I, T, and ABD genomes. The DNA probe hybridized with PalI- or TaqI-digested total DNAs from Thinopyrum elongatum (JeJe diploid) and T. elongatum x Psathyrostachys juncea (JeN hybrid), but not with those from L. arenarius (NNNNXXXX octoploid) and all tetraploid Leymus species (NNXX). Attempts to cross diploid Thinopyrum and tetraploid Leymus species yielded only one triploid hybrid, T. elongatum x L. salinus ssp. salmonis. Meiotic chromosome associations at metaphase I of pollen mother cells in the triploid hybrid averaged 19.69 univalents, 0.64 bivalents, and 0.01 trivalents per cell. Chromosome pairings in the tetraploid hybrids of L. mollis x L. salinus ssp. salmonis, and the reciprocal cross, indicate that L. mollis and L. salinus ssp. salmonis shae the same genomic constitution. Both the DNA probe and genome analysis results confirm the absence of the J genome in the seven additional Leymus species tested. Meiotic data indicated that tetraploid Leymus species could not have the genome formula N1N1N2N2; thus their genome formulas should remain as NNXX until the source of X is identified.     

Festuca inops and Festuca gracilior (Poaceae): are they two different species?

According to current systematics, Festuca inops and F. gracilior are two distinct species. However, they are hardly distinguishable from each other on the basis of their morphological characters. Festuca inops is considered a diploid species endemic to Italy, while F. gracilior has a discontinuous distribution area, apparently related to chromosomal levels: diploid populations in Italy and south-east France, tetraploid populations in north-east Spain. The diploid populations of both taxa from Italy and south-east France are investigated in the present study. Nearly 1000 exsiccata were examined and morphometric analysis was carried out on macro- and micromorphological features of 119 specimens (including type-specimens) and on 20 natural populations (including loci classici ). All these data showed that the two species should be referred to a single taxon, for which the rank of species seems to be appropriate. This result is supported by karyological, ecological and chorological data and was confirmed by the results of ISSR analysis. According to nomenclatural rules, the legitimate name for the species is Festuca inops De Not.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 151 , 239–258.     

GISH/FISH mapping of genes for freezing tolerance transferred from Festuca pratensis to Lolium multiflorum

The first backcross breeding programme for the transfer of freezing-tolerance genes from winter hardy Festuca pratensis to winter-sensitive Lolium multiflorum is described. A partly fertile, triploid F(1) hybrid F. pratensis (2n=2x=14) x L. multiflorum (2n=4x=28) was employed initially, and after two backcrosses to L. multiflorum (2x) a total of 242 backcross two (BC(2)) plants were generated. Genomic in situ hybridisation (GISH) was performed on 61 BC(2) plants selected for their good growth and winter survival characters in the spring following one Polish winter (2000-2001). Among the winter survivors, diploid chromosome numbers were present in 80% of plants. An appropriate single Festuca introgression in an otherwise undisturbed Lolium genome could provide increased freezing tolerance without compromise to the good growth and plant vigour found in Lolium. Among all the diploids, a total of 20 individuals were identified, each with a single F. pratensis chromosome segment. Another diploid plant contained 13 Lolium chromosomes and a large metacentric F. pratensis chromosome, identified as chromosome 4, with two large distal Lolium introgressions on each chromosome arm. Three of the diploid BC(2), including the genotype with Festuca chromosome 4 DNA sequences, were found to have freezing tolerance in excess of that of L. multiflorum, and in one case in excess of the F. pratensis used as control. A detailed cytological analysis combining GISH and fluorescence in situ hybridisation analyses with rDNA probes revealed that the other two freezing-tolerant genotypes carried a Festuca chromosome segment at the same terminal location on the non-satellite arm of Lolium chromosome 2.     

Rare recent natural hybridization in Hieracium s. str. – evidence from morphology, allozymes and chloroplast DNA

The first proven data on natural hybridization in the genus Hieracium s. str. are presented. Plants with intermediate morphological characters between the diploids H. alpinum and H. transsilvanicum were found in the Muntii Rodnei (Romanian Eastern Carpathians) in 2001 and in the Chornohora Mts (Ukrainian Eastern Carpathians) in 2003. While plants of intermediate morphology between usually so called basic species are usually tri- or tetraploid in Hieracium s. str., these plants were diploid (2n=18) like both parental species in this region. The Romanian plant did not produce fertile achenes in free pollination and in control backcrosses with H. transsilvanicum, two hybrids from Ukraine were completly seed sterile in free pollination and reciprocal crosses. Pollen stainability as an indirect measure of male fertility was quite high in the studied Ukrainian hybrid plants and similar to the parental taxa. Evidence from allozyme analysis also confirmed the hybrid origin of the studied plants. Sequencing and PCR-RFLP analyses of the trnT-trnL intergenic spacer revealed that all hybrid plants had the H. transsilvanicum chloroplast DNA haplotype. Maternal inheritance of chloroplast DNA in this particular cross was proved with artificial hybrids from reciprocal experimental crosses between H. alpinum and H. transsilvanicum. In both localities, the natural hybrid plants were found in disturbed habitats, exceptionally allowing contact of the otherwise ecologically vicariate parental species. Morphologically, the hybrid plants belong to H. × krasani Woł.     

Fitness differences among diploids, tetraploids, and their triploid progeny in Chamerion angustifolium: mechanisms of inviability and implications for polyploid evolution

Abstract. Theoretical models indicate that the evolution of tetraploids in diploid populations will depend on both the relative fitness of the tetraploid and that of the diploid-tetraploid hybrids. Hybrids are believed to have lower fitness due to imbalances in either the ploidy (endosperm imbalance) or the ratio of maternal to paternal genomes in their endosperm (genomic imprinting). In this study we created diploids, tetraploids, and hybrid triploids of Chamerion angustifolium from crosses between field-collected diploid and tetraploid plants and evaluated them at six life stages in a greenhouse comparison. Diploid offspring (from 2 x × 2 x crosses) had significantly higher seed production and lower biomass than tetraploid offspring (from 4 x × 4 x crosses). Relative to the diploid, the cumulative fitness of tetraploids was 0.67. In general, triploids (from 2 x × 4 x , 4 x × 2 x crosses) had significantly lower seed production, lower pollen viability, and higher biomass than diploid individuals. Triploid offspring derived from diploid maternal parents had lower germination rates, but higher pollen production than those with tetraploid mothers. Relative to diploids, the cumulative fitness of 2 x × 4 x triploids and 4 x × 2 x triploids was 0.12 and 0.06, respectively, providing some support for effect of differing maternal:paternal ratios and endosperm development as a mechanism of hybrid inviability. Collectively, the data show that tetraploids exhibit an inherent fitness disadvantage, although the partial viability and fertility of triploids may help to reduce the barrier to tetraploid establishment in sympatric populations.     

Allozyme and morphological observations on Festuca hyperborea, compared with F. baffinensis and F. brachyphylia (Poaceae) from the Canadian Arctic

Phenotypic plasticity in the size of leaf cross sections and in the visibility of trichomes on the leaf abaxial surface for a plant of Festuca hyperborea grown in different microhabitats is illustrated. Isozyme analyses indicate that tetraploid F. hyperborea from the Canadian Arctic is not likely a variant of tetraploid F. baffinensis and probably not a recent ancestor of the hexaploid F. brachyphylia . Although 10 populations were collected in the Canadian Arctic Archipelago as morphologically intermediate between Festuca species, with F. hyperborea considered a possible parent, no isozyme evidence suggested that this species hybridizes with either F. baffinensis or F. brachyphylia . Within plants sampled as F. hyperborea , 12 isozymatically distinct plants and one hybrid plant between this group and F. hyperborea were found.