Haploidy from Hordeum interspecific crosses

Summary Interspecific crosses of Hordeum brachyantherum (2n = 28) and H. depressum (2n = 28) with H. bulbosum (2n = 14 or 28) and H. vulgare (2n = 14 or 28) were made. Crosses between brachyantherum and diploid bulbosum resulted in dihaploids (2n = 14) of brachyantherum and hybrids (2n = 21), whilst the crosses of brachyantherum by tetraploid bulbosum or vulgare gave hybrid progeny. Similarly, crosses between H. depressum and diploid bulbosum resulted in dihaploids (2n = 14) of depressum and hybrids (2n = 21), whereas depressum by tetraploid bulbosum or vulgare invariably produced hybrids.Cytological observations on 12 day old embryos obtained from these crosses revealed chromosome variability down to 14 in crosses with diploid bulbosum indicating thereby that chromosome elimination leads to haploid formation. Embryonic cells from the brachyantherum by diploid vulgare cross also exhibited a certain degree of chromosomal instability as micronuclei.The results indicate that the ratio of parental genomes in the zygote determines whether haploids or hybrids will be produced in crosses of brachyantherum or depressum with bulbosum. Furthermore, brachyantherum appears to be more efficient in eliminating bulbosum chromosomes in comparison with depressum.     

Chromosome numbers and taxonomic implications in the fern genusAzolla (Azollaceae)

Investigation of chromosome numbers of allAzolla species, and for the first time of hybrids, has been undertaken. Removal of wax from the leaf surface proved invaluable in achieving clear cytological preparations and providing unambiguous chromosome numbers. In contrast to previous records, the speciesA. pinnata, A. filiculoides, A. filiculoides var.rubra, A. caroliniana, A. microphylla, andA. mexicana were found to be 2n=44, andA. nilotica to be 2n=52. Several triploids (2n=66) and one tetraploid (2n=88) were identified. No geographical pattern could be observed in the distribution of triploids which probably derive from the function of unreduced gametes. The chromosome number of hybrids occasionally deviates from the diploid chromosome number (2n=44). The small chromosome size limits karyotypic analysis and only differences in overall chromosome size can be observed. Taxonomic implications of chromosome numbers and sizes are discussed.     

Cytogenetic and phytochemical investigations in hybrids of MacaronesianDryopteris (Pteridophyta: Aspidiaceae)

The cytology and phloroglucinol composition of wild and synthesized hybrids of MacaronesianDryopteris species have been examined. Both cytological and chemical evidence suggest that the tetraploidD. guanchica has arisen from the diploid speciesD. aemula andD. maderensis by hybridization and chromosome doubling. However, the formation of the characteristic fully aromatic compounds inD. aemula appears to be suppressed in most of the hybrids involving this species, and inD. guanchica.     

Hybridization studies in the genusSilene sectt.Siphonomorpha andAuriculatae (Caryophyllaceae)

Ten species in the genusSilene sectt.Siphonomorpha andAuriculatae were crossed artificially involving 612 crosses to test inter- and infraspecific, intervarietal and intersectional crossability. In sect.Siphonomorpha all interspecific crosses (between diploids) failed due to cross- or seed-incompatibility; however, intervarietal crosses betweenS. gigantea var.gigantea andS. gigantea var.incana produced hybrids. In sect.Auriculatae hybrids were produced betweenS. vallesia andS. boryi, both tetraploids, but crosses between these and the diploid species were unsuccessful. The delimitation and status of the species in both sections was supported by the crossing results.     

Hybridization and speciation in the genusDryopteris (Pteridophyta:Dryopteridaceae) on Pico Island in the Azores

The cytology of wild hybrids ofDryopteris from Pico, Azores has been examined. This has confirmed the parentage of the hybrids, and provided evidence to support the view that the tetraploid speciesD. crispifolia has evolved in the Azores from two diploid species,D. aemula andD. azorica, by hybridization and polyploidy.     

A genetic contribution to the taxonomy ofOenothera sect.Oenothera (including subsectionsEuoenothera,Emersonia, Raimannia andMunzia)

Based on an analysis of results from experimental hybridization, the plants assigned byMunz toOenothera subg.Oenothera and subg.Raimannia, now divided into approximately 76 species, are referred to a single section,Oenothera. This section is in turn divided into five subsections:Euoenothera, Munzia, Raimannia, Emersonia, and an undescribed group of three species related toOenothera pubescens. Euoenothera is maintained in the traditional sense, and includes about 14 species of North America, widely naturalized elsewhere.Munzia consists of 45 species, comprising three series, and native to South America.Raimannia is restricted to a group of approximately 11 North American species.Emersonia comprises four rather heterogenous species of northern Mexico and southern New Mexico, of whichOenothera macrosceles, O. maysillesii, andO. organensis have been described. Within these four subsections, interspecific hybrids can be made in general, although plastid differentiation often leads to incompatibilities. With varying degrees of difficulty, hybrids were produced in all intersectional combinations involvingEuoenothera, Emersonia, Munzia, andRaimannia, the most difficult being those betweenEmersonia andRaimannia. Based on their habit and distribution,Emersonia species, and especiallyOenothera maysillesii, appear to resemble most closely the common ancestor of the section,Euoenothera andMunzia to have been derived from it or its common ancestor, andRaimannia perhaps to be more closely related to the phylogenetic branch that leads toEuoenothera.     

Chromosomal evolution inEpilobium sect.Epilobium (Onagraceae)

WithinEpilobium sect.Epilobium, a cytological analysis of 121 experimental hybrids, involving 40 species, indicates the presence of a widespread BB chromosome arrangement in Eurasia, Africa, and Australasia, as well as in North and South America less commonly. The AA chromosome arrangement, which differs from BB by one reciprocal translocation, occurs in North America, South America, and in at least three European species. The CC arrangement, which differs from AA by two reciprocal translocations, characterizes theAlpinae, a circumboreal group. Distinctive or only partly worked out chromosome arrangements occur in the EuropeanE. duriaei andE. nutans and in the North AmericanE. luteum, E. obcordatum, E. oregonense, andE. rigidum. With earlier results, the chromosome arrangements of some 65 of the estimated 185 species of the section have been established fully or partly.     

DNA evidence for multiple origins of intergeneric allopolyploids in annualMicroseris (Asteraceae)

Seventy populations of North American annualMicroseris, Stebbinsoseris, andUropappus species were examined for chloroplast and nuclear ribosomal DNA restriction site variability to determine the origin of the allotetraploid speciesS. heterocarpa andS. decipiens. Previously identified chloroplast DNA restriction site variants were used in concert with restriction site variation forNco I in the nuclear-encoded ribosomal DNA repeat. The presence of two, mutually exclusive restriction site gains were observed in diploid populations ofM. douglasii; these same variants were also found in populations of allotetraploidS. heterocarpa, indicating mutiple origins of this species from different maternal diploid populations ofM. douglasii. Variation in the rDNA repeat between the diploid annual species and the putative paternal genome ofU. lindleyi was found to be additive inS. heterocarpa. A similar relationship was observed for the origin ofS. decipiens; cpDNA restriction site variants found inM. bigelovii andM. douglasii were present inS. decipiens. The rDNANco I variants also were additive in this purported allotetraploid. These results confirm the reticulate evolutionary pattern inStebbinsoseris and provide another example of multiple origins of intergeneric allopolyploids.     

Differentiation of fourHordeum sect.Stenostachys spp. (Poaceae: Triticeae) using multivariate morphometrics

Field collections and 296 herbarium sheets were examined for 27 morphometric variables. A priori species identifcation was based on geographical distribution except forH. californicum, a diploid species primarily occurring in California and differing from the much more widespread tetraploidH. brachyantherum that thrives in N. America and N.E. Asia;H. capense grows in S. Africa andH. secalinum mainly in Europe. Various cluster analyses were used followed by cluster recovery verification. Classificatory discriminant analysis and validation by the bootstrap yielded 85–90% overall total correct classification of the four species. Canonical analysis revealed thatH. californicum occupies an intermediate phenetic position among the other three distinct species. Factors of shape differences were unravelled and portrayed by shearing. A revised key to species was drawn up.     

Crossing experiments of lettuce cultivars and species (Lactuca sect.Lactuca,Compositae)

The degree of relationships withinLactuca sativa and three wild relativesL. serriola, L. saligna, andL. virosa was studied by observing the performance, vigour and fertility of the F 1 hybrids obtained from crosses made in and between the four species. The crosses ofL. saligna ×L. virosa and the reciprocal crosses produced no hybrids.L. saligna andL. virosa are the least related of the four species.L. sativa ×L. serriola and the reciprocal crosses were successful and produced fertile hybrids These two species are genetically very closely related.L. saligna is known to produce, as a female parent, hybrids withL. sativa andL. serriola. Now the reciprocal cross was successful for the first time, so the unability to obtain hybrids in the past was based on the choice of accessions and not caused by unilateral incompatibility.L. virosa ×L. sativa and the reciprocal combination produced hybrids. The combinationL. serriola ×L. virosa produced hybrids with very limited fertility. In contrast to earlier reports (sterile hybrids) one combination of the reciprocal cross too produced hybrids with very limited fertility.—Some of theL. saligna ×L. sativa (and reciprocal) hybrids were found to look strikingly likeL. serriola. This adds evidence for the descent ofL. serriola andL. sativa:L. saligna also made part of the ancestral complex of the cultivated lettuce.     

Experimental hybridization between catfishes of the families Clariidae and Pangasiidae in Thailand

Synopsis Hybrids between the clariid speciesClarias macrocephalus andC. batrachus and the pangasiid speciesPangasius sutchi were obtained by hormone injection of brood stock and artificial fertilization. Pure parental crosses as well as all possible hybrid combinations were obtained. Fertility, hatchability, and post-yolk absorption survival was high (66–99%) in all pure parental crosses and in all crosses between the two species ofClarias. In crosses betweenClarias andPangasius fertilization was also very high (68–97%) but hatchability varied from 11 to 23% and post-yolk absorption survival from 0% inPangasius sutchi ×Clarias batrachus to about 50% inPangasius sutchi ×Clarias macrocephalus . The longest-lived hybrids ofPangasius andClarias were those ofPangasius sutchi ×Clarias macrocephalus which survived until the experiment was prematurely terminated due to contaminated food after 4 1/2 months, at which point they had grown to total lengths of 8–14cm. The hybrids comprised four morphotypes, two relativelyPangasius-like and two relativelyClarias-like, but all markedly different from the parental species.     

Improved isolation of dihaploidsolanum tubersoum protoplasts and the production of somatic hybrids between dihaploidS. tuberosum andS. brevidens

Summary A modified protoplast isolation technique, applicable to a range of dihaploidSolanum tuberosum genotypes, has been developed. A combination of high calcium and high pH was used to fuse mesophyl protoplasts of dihaploidS. tuberosum (PDH40) and the diploid wild speciesS. brevidens. Large numbers of colonies were obtained after fusion and putative hybrids selected on the basis of phenotype from regenerated shoots. From these, 11 somatic hybrid plants have been identified by their isoenzyme patterns and morphologic characteristics. Four of these hybrids had the expected chromosome number of 48. The approach of mass culture after fusion followed by selection of hybrids from regenerated shoots and the application of somatic hybridization to potato breeding are discussed.     

Intergeneric hybridization and C-banding patterns inHordelymus (Triticeae,Poaceae)

Crosses ofHordelymus europaeus (2n = 4x = 28) with four genera in theTriticeae were attempted. Adult hybrids were obtained in combinations withHordeum bogdanii (2x),Hordeum depressum (4x), andSecale cereale (2x). The meiotic pairing was very low in the hybrids withH. bogdanii andSecale cereale (0.12 and 0.30 chiasmata/cell, respectively), whereas high pairing (9.90 chiasmata/cell) was found in hybrids withH. depressum due to autosyndetic pairing ofH. depressum chromosomes. The chromosome complement ofHordelymus europaeus comprised 16 metacentrics, 8 submetacentrics, and 4 SAT-chromosomes. The Giemsa C-banding patterns of the chromosomes were characterized by small to minute bands at no preferential positions. It is hypothesized thatHordelymus europaeus may genomically be closest related toTaeniatherum andPsathyrostachys spp.     

Suitability of AFLP markers for the study of genomic relationships within theOxalis tuberosa alliance

O. tuberosa is an Andean crop that belongs to the worldwide distributed genusOxalis. On the basis of their chromosome numbers the following species (O. herrerae, O. lotoides, O. medicaginea, O. mollissima, O. oblongiformis, O. peduncularis, O. spiralis, O. subintegra, O. tabaconanensis, O. tuberosa, O. villosula) were placed in an alliance. To analyse five species belonging to theOxalis tuberosa alliance (O. oblongiformis, O. peduncularis, O. tabaconanensis, O. tuberosa andO. villosula) and a distant member of the genus (O. articulata), we examined 253 AFLP markers generated after amplification using four primer combinations. Within the alliance, two main clusters were observed, one containing the diploid species and the other group with the polyploid speciesO. tuberosa. All of the primer combinations assayed showed the same clustering pattern. Grouping of accessions of each species by data analysis corresponded largely with their previous taxonomic classifications. The concordance between the clustering of the individuals belonging to different species obtained in this work show the appropriateness of AFLP markers for this type of study. The results obtained are in good agreement with the cytogenetic hypothesis and showed a clustering behaviour, which is similar to the one previously obtained using ITS rDNA nucleotide sequence comparison.     

The influence of parental genotype on the chromosome behaviour of Hordeum vulgare X H. bulbosum diploid hybrids

Summary The PMCs of 74 diploid hybrids involving ten H. vulgare varieties and three H. bulbosum lines were analysed at metaphase I and chromosome number and chiasma frequency recorded. There were differences between parental combinations and between plants within those combinations for both chromosome and chiasma number. It is suggested that these characters are controlled by both parents and that differences between plants within families reflect the heterozygosity of the H. bulbosum parents. Chromosomally stable, high pairing lines have been identified for use in a backcrossing programme to introduce H. bulbosum characters to the H. vulgare germplasm.     

A natural hybrid ofMonophyllaea (Gesneriaceae) in the tropical rain forests of West Sumatra

A natural hybrid (2n = 21) between the parapatric rain forest speciesMonophyllaea hirtella (2n = 20) andM. horsfieldii (2n = 22) (Gesneriaceae) has been observed at Sg. Lubuk Paraku, Padang, W. Sumatra. The hybrids showed intermediary characters between the parental species in the inflorescence structure, flower size and colour, indumentum, chromosome numbers, and habitat. The 29% pollen fertility of a single hybrid specimen suggests that the genetical isolation between the parental species may break down and reproduction over subsequent generations may occur.     

Comparisons of the hybrids Hordeum chilensexH. vulgare,H. chilensexH. bulbosum,H. chilensexSecale cereale and the amphidiploid of H. chilensexH. vulgare

Summary Diploid hybrids between Hordeum chilense and three other species, namely H. vulgare, H. bulbosum and Secale cereale, are described together with the amphidiploid of H. chilensexH. vulgare. Both the diploid hybrid and the amphidiploid of H. chilensexH. vulgare were chromosomally unstable, H. chilensexH. bulbosum was less so, while H. chilensexS. cereale was stable. Differential amphiplasty was found in all combinations. No homoeologous pairing was found in the Hordeum hybrids but in H. chilensexS. cereale there was chromosome pairing both within the two genomes and between the genomes.     

Investigations into the evolutionary history of the polyploid complexAntennaria neodioica (Asteraceae: Inuleae)

TheAntennaria neodioica polyploid agamic complex is a polymorphic species occurring across North America mainly north of the terminal margin of the Wisconsin glacier. This taxonomically difficult group has recently been treated as consisting of the four subspeciesA. neodioica subsp.canadensis, subsp.howellii, subsp.neodioica, and subsp.petaloidea. TheA. neodioica agamic complex has been considered of hybrid origin with several sexual diploid species constituting its parentage. Crosses were made among five sexual diploid species ofAntennaria, morphologically similar toA. neodioica s.l., in an attempt to discover its origins. Representative specimens of the five diploid species,A. neodioica s. l., and the synthetic interspecific F₁ hybrids were subjected to various analyses including PCA, cluster (UPGMA), and discriminant analyses. Results suggest that theA. neodioica complex is of multiple hybrid origin involving the four diploid speciesA. neglecta, A. plantaginifolia, A. racemosa, andA. virginica. BecauseA. neodioica is the result of diverse origin it is more desirable to consider the agamic complex as a separate, distinct species from its sexual diploid relatives. Several morphological characters in the diploid species were determined to be polygenically inherited.Investigations into the Evolutionary History of the Polyploid Complexes inAntennaria (Asteraceae: Inuleae). I.     

Characterisation of the double genome structure of modern sugarcane cultivars (Saccharum spp.) by molecular cytogenetics

Cultivated sugarcane clones (Saccharum spp., 2n=100 to 130) are derived from complex interspecific hybridizations between the speciesS. officinarum andS. spontaneum. Using comparative genomic DNA in situ hybridization, we demonstrated that it is possible to distinguish the chromosomes contributed by these two species in an interspecific F1 hybrid and a cultivated clone, R570. In the interspecific F1 studied, we observed n+n transmission of the parental chromosomes instead of the peculiar 2n+n transmission usually described in such crosses. Among the chromosomes of cultivar R570 (2n=107–115) about 10% were identified as originating fromS. spontaneum and about 10% were identified as recombinant chromosomes between the two speciesS. officinarum andS. spontaneum. This demonstrated for the first time the occurrence of recombination between the chromosomes of these two species. The rDNA sites were located by in situ hybridization in these two species and the cultivar R570. This supported different basic chromosome numbers and chromosome structural differences between the two species and provided a first bridge between physical and genetical mapping in sugarcane.     

Stylosanthes sp. aff.S. scabra: a putative diploid progenitor ofStylosanthes scabra (Fabaceae)

Stylosanthes sp. aff.S. scabra is an undescribed taxon showing affinities with the allotetraploid speciesS. scabra, but distinct in a number of attibutes. Several collections show potential as forage for clay soils in northern Australia. Twelve accessions have been analysed using STS (sequence-tagged-sites) as genetic markers, and they all displayed STS phenotypes of typical diploid species. Taking into account their morphological similarities, the STS analysis provides strong evidence thatStylosanthes sp. aff.S. scabra might be a diploid progenitor of the allotetraploidS. scabra. This speculation was supported by cytological examinations. Somatic chromosome numbers of two of these accessions were counted and both were found to be diploid (2n = 20). The level of polymorphism among the 12Stylosanthes sp. aff.S. scabra accessions, estimated using randomly amplified polymorphic DNA (RAPD) as markers, was 7.8%, and the dissimilarity value betweenStylosanthes sp. aff.S. scabra andS. viscosa (the other putative progenitor ofS. scabra) was 89%.