Effect of exogenous plant growth regulators on in vitro seed growth,embryo development,and haploid production in a cross between H. vulgare (L.) and H. bulbosum (L.)

Exogenous plant growth regulators are known to increase the efficiency of interspecific and intergeneric crosses. In vitro floret culture provides a defined system for assessing the importance of various plant growth regulators on the determinants of haploid production efficiency (seed set, embryos per seeds, and plants per embryos) in Hordeum vulgare × Hordeum bulbosum crosses. The individual and combined effects of three plant growth regulators (2,4-D, GA₃ and kinetin) on in vitro seed growth, embryo development and haploid production efficiency were tested in floret culture of the cross H. vulgare, cultivar Klages × H. bulbosum. All treatments, except kinetin alone, produced larger seeds and more embryos/100 seeds than the control (no plant growth regulator). 2,4-D alone was superior to GA₃ alone in haploid production efficiency (70.6 vs. 51.5) as measured by the number of plants regenerated/100 florets pollinated. Although kinetin +2,4-D+GA₃ produced the largest seeds and embryos, no advantage over 2,4-D alone was observed in haploid production efficiency. 2,4-D alone or kinetin +2,4-D are recommended for the purpose of barley haploid production in floret culture using the bulbosum method.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid     

A taxonomic investigation of theHordeum murinum Complex (Poaceae)

The purpose of this paper is to produce a classification of theHordeum murinum complex. Using an experimental design to account for the environmental component of the phenotypic variation, material from sources throughout the old-world range ofH. murinum was grown in a common environment. Eight morphological characters were measured on the resulting triplets (i.e., the central and lateral florets, glumes and rachis internode). Clustering, variance and discriminants analyses of these data suggest that there are three distinct groups. This classification is then examined for consistency with hypotheses obtained from the work of other researchers and discussed in light of the known biology of the plants. An identification key to members of the complex using the triplet characters is provided.     

Giemsa C-banded karyotypes of diploid and tetraploidHordeum bulbosum (Poaceae)

The similar-looking basic genomes ofHordeum bulbosum (2x and 4x) have five rather similar metacentric, one submetacentric, and one satellited choromosome. C-banding patterns are characterized by one or two centromeric, or juxtacentromeric, small to larger bands in most chromosomes, by bands at the nucleolar organizers, by small or very small telomeric bands, and by the nearly complete lack of intercalary bands. Banding pattern polymorphism is widespread. Banding patterns supported by chromosome morphology enable identification of homologues, and discrimination between non-homologues inH. bulbosum (2x). The C-banded karyotype ofH. bulbosum (4x) supports an autopolyploid origin, but it was possible to identify only homologues of submetacentrics and SAT-chromosomes.     

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 taxometric analysis of seed proteins in the genusBriza s. l. (Poaceae)

The species ofBriza L. s. l. are disjunctly distributed between Eurasia and South America. A taxometric analysis has been carried out, using data from electrophoresis of seed polypeptides and published morphological characters. Evidence from a cluster analysis of seed polypeptide data strongly supports the recognition of two different natural groups, a S. American group and a Eurasian group, and is consistent with karyotype data, but does not support the generic groupings of South American taxa derived from the morphological data.     

Monosomic and double monosomic substitutions of Hordeum bulbosum L. chromosomes into H. vulgare L.

Summary One of the aims of the interspecific crossing programme between Hordeum vulgare L. and H. bulbosum L. has been to introgress desirable genes into barley from the wild species. However, despite their close taxonomic relationship there have been few reports of achieving this objective using amphidiploid hybrids. In order to broaden the range of available hybrids, partially fertile triploids between H. vulgare (2n = 2x = 14) and H. bulbosum (2n = 4x = 28) were developed and crossed with H. vulgare as female parent. From 580 progeny which were screened, eight putative single monosomic chromosome substitution lines and one double monosomic substitution were identified by cytological analysis. These involved the substitution of H. vulgare chromosome 1 (two lines), 6 (four lines), 6L (one line), 7 (one line) and 1 + 4 (one line) by their respective H. bulbosum homoeologues. The H. bulbosum chromosome was frequently eliminated during plant development, but it was observed regularly in pollen mother cells of two lines. However, pairing between the H. bulbosum chromosome and its H. vulgare homoeologue was low. Several of the lines were more resistant than their H. vulgare parents to powdery mildew (Erysiphe graminis DC. f.sp. hordei Em. Marchai), net blotch (Drechslera teres Sacc.) and scald (Rhynchosporium secalis (Oudem.) Davis). Apart from their use in studying genome relationships, their value to plant breeders will depend on the ease of inducing translocations between the parental chromosomes.     

Hordein polymorphism in diploid and tetraploid Mediterranean populations of the Hordeum murinum L. complex

Polymorphism analyses of the hordeins, main storage proteins in barley, were conducted on 35 natural populations of Hordeum murinum s.l. from North Africa; this specific complex includes three subspecies with two ploidy levels: H. murinum subsp. glaucum (2n=2x=14), H. murinum subsp. leporinum and subsp. murinum (2n=4x=28). Twenty of these populations belong to the diploid subsp. glaucum, 14 other tetraploid populations belong to the subsp. leporinum. In addition, six populations of the tetraploid murinum were sampled in France: two along the Mediterranean coast and four in Brittany. The polymorphism observed in the electrophoretic patterns highlights strong correlations between bioclimatic features and di- and tetraploid taxa distribution. Moreover, the variation was not randomly distributed within the different ploidy levels, and is correlated with environmental factors. The ecological differentiation of the two main taxa, H. murinum subsp. leporinum and subsp. glaucum is clearly highlighted.     

Transfer of a dominant gene for powdery mildew resistance and DNA from Hordeum bulbosum into cultivated barley (H. vulgare)

Summary In an attempt to transfer traits of agronomic importance from H. bulbosum into H. vulgare we carried out crosses between four diploid barley cultivars and a tetraploid H. bulbosum. Eleven viable triploid F₁ plants were produced by means of embryo rescue techniques. Meiotic pairing between H. vulgare and H. bulbosum chromosomes was evidenced by the formation of trivalents at a mean frequency of 1.3 with a maximum of five per cell. The resulting triploid hybrids were backcrossed to diploid barley, and nine DC₁ plants were obtained. Three of the BC₁ plants exhibited H. bulbosum DNA or disease resistance. A species specific 611-bp DNA probe, pSc119.2, located in telomeres of the H. bulbosum genome, clearly detected five H. bulbosum DNA fragments of about 2.1, 2.4, 3.4, 4.0 and 4.8 kb in size present in one of the BC₁ plants (BC₁-5) in BamHI-digésted genomic Southern blots. Plant BC₁-5 also contained a heterozygous chromosomal interchange involving chromosomes 3 and 4 as identified by N-banding. One of the two translocated chromosomes had the H. bulbosum sequence in the telomeric region as detected using in situ hybridization with pSc119.2. Two other BC₁ plants (BC₁-1 and BC₁-2) were resistant to the powdery mildew isolates to which the barley cultivars were susceptible. Seventy-nine BC₂ plants from plant BC₁-2 segregated 32 mildew resistant to 47 susceptible, which fits a ratio of 11, indicating that the transferred resistance was conditioned by a single dominant gene. Reciprocal crosses showed a tendency towards gametoselection that was relative to the resistance. Mildew resistant plant BC₁-2 also had a 1-kb H. bulbosum DNA fragment identified with a ten-base random primer using polymerase chain reaction (PCR). Forty-three BC₁ plants, randomly sampled from the 79 BC₁ plants, also segregated 2320 for the presence versus absence of this 1-kb H. bulbosum DNA fragment, thereby fitting a 11 ratio and indicating that the PCR product originated from a single locus. The 1-kb DNA fragment and disease resistance were independently inherited as detected by PCR analysis of bulked DNA from 17 resistant and 17 susceptible plants as well as by trait segregation in the 43 individual plants. The progenies produced could serve as an important resistant source in plant breeding. This is the first conclusive report of the stable transfer of disease resistance and DNA from H. bulbosum to H. vulgare.     

Variability and structure of natural populations of Hordeum murinum L. based on morphology

Based on a detailed statistical analysis of 19 quantitative and four qualitative characters of 31 samples of Hordeum murinum from Poland, it was stated that the small variability of these characters does not permit to distinguish any intra-specific units. Only a slight geographical differentiation of populations was found. Based on the differences discovered, the populations have been divided into two groups: one with the centre of distribution in the North-East (group E) and the second in South-West Poland (group W). This differentiation might result from the different routes of migration of this species into Poland. The chromosome number (2n = 28) confirms earlier reports from Poland. New data concerning the distribution of H. murinum in north-eastern Poland are also presented. A detailed study on the distribution of H. murinum in Poland is under way. It is the fourth paper from the series: Biodiversity of wild Triticeae (Poaceae) in Poland. The first is: Mizianty M. (2005). Variability and structure of natural populations of Elymus caninus (L.) L. based on morphology. Pl. Syst. Evol 251: 199–216, the second: Mizianty M. et al. (2006). Variability and structure of natural populations of Elymus caninus (L.) L. and their possible relationship with Hordelymus europaeus (L.) Jess. ex Harz as revealed by AFLP analysis. Pl. Syst. Evol. 256: 193–200, the third: Mizianty M. and Szklarczyk M. (2005). Systematic significance of Elymus caninus morphological characters revealed by AFLP analysis. In: Frey L. (red.) Biology of grasses. W. Szafer Institute of Botany, Polish Academy of Sciences. Kraków, pp. 9-21.     

Characterisation of progeny from backcrosses of triploid hybrids between Hordeum vulgare L. (2x) and H. bulbosum L (4x) to H. vulgare

Interspecific hybridisations between Hordeum vulgare L. (cultivated barley) and H. bulbosum L. (bulbous barley grass) have been carried out to transfer desirable traits, such as disease resistance, from the wild species into barley. In this paper we report the results of an extensive backcrossing programme of triploid hybrids (H. vulgare 2x x H. bulbosum 4x) to two cultivars of H. vulgare. Progenies were characterised cytologically and by restriction fragment length polymorphism analysis and comprised (1) haploid and diploid H. vulgare plants, (2) hybrids and aneuploids, (3) single and double monosomic substitutions of H. bulbosum chromosomes into H. vulgare and (4) chromosomal rearrangements and recombinants. Five out of the seven possible single monosomic chromosome substitutions have now been identified amongst backcross progeny and will be valuable for directed gene introgression and genome homoeology studies. The presence amongst progeny of 1 plant with an H. vulgare-H. bulbosum translocated chromosome and one recombinant indicates the value of fertile triploid hybrids for interspecific gene introgression.     

Taxonomical investigations onHelleborus viridis s. l. (Ranunculaceae) in Northern Italy

The taxon commonly namedHelleborus viridis in Lombardy (NW. Italy) differs from both, subsp.viridis and subsp.occidentalis, and is shown to fall within the variation range ofH. odorus subsp.laxus which has been reported so far for NE. Italy only. TrueH. viridis within Italy grows only in the Maritime Alps.     

Comparative responses of tetraploid wheats pollinated with Zea mays L. and Hordeum bulbosum L.

Ten different tetraploid wheat (Triticum turgidum) genotypes were pollinated with maize (Zea mays). Fertilization was achieved in all ten genotypes and no significant difference in fertilization frequency between the tetraploid wheat genotypes was detected. A mean of 41.1% of pollinated ovaries contained an embryo. All these crosses were characterized by the elimination of the maize chromosomes, and the resulting embryos were haploids. Six of the tetraploid wheat genotypes were also pollinated with Hordeum bulbosum. Fertilization frequencies with H. bulbosum were much lower (mean=13.4%), and significant differences between the tetraploid wheat genotypes were detected. Observation of pollen tube growth revealed that part of the incompatibility reaction between tetraploid wheats and H. bulbosum was due to an effect similar to that of the Kr genes, namely pollen tube growth inhibition. These results indicate that pollinations with maize may have potential as a broad spectrum haploid production system for tetraploid wheats. Present address: Agriculture Canada, Research Branch, Central Experimental Farm, Bldg 50, Ohawa, Ontario, Canada K1A OC6     

Gynoecium and perianth inZanthoxylum s.l. (Rutaceae)

The question whether the uniseriate perianth ofZanthoxylum L. s. str. is homologous with the calyx or the corolla of taxa included inFagara, or of an independent origin, has been controversial for a long time, but the arguments mostly have remained theoretical. The present investigation of floral structures indicates that there are two different types of uniseriate perianth inZanthoxylum s. str. Therefore, this taxon does not represent a natural group and should be united withFagara asZanthoxylum s.l. The infrageneric taxonomy of this genus is still very ambiguous. It is shown that differences in indumentum, number of sepals and petals (5-4-3) resp. perianth segments (4–9), stamens (3–6), and free carpels (1–5) are of systematic relevance. Particularly important but so far neglected is carpel shape, where an acrostylous and an anacrostylous-basistylous type can be recognized. Stigmata of 2 or more carpels mostly fuse to form a compitum. 4–5-merous flowers with calyx and corolla, and acrostylous carpels are considered as plesiomorphic character states in the genus. On the basis of ± corresponding morphological and phytochemical progressions a working hypothesis about the relationships withinZanthoxylum s.l. is presented in graphical form (Fig. 9).Adapted from a lecture held at the 10th Symposion on Morphology, Anatomy, and Systematics in Göttingen, February 1991.     

Biochemical variation to determine phylogenetic relationships betweenHordeum chilense and other American species of the genusHordeum (Poaceae)

Slab gel electrophoresis techniques have been applied to the study of isozyme and kernel protein patterns in 20 accessions ofHordeum chilense and related species in order to elucidate their phylogenetic relationships. On the basis of our results we can conclude that: (1) Conventional classification based on morphological characters does not totally agree with biochemical data. (2) Sectt.Anisolepis andCritesion seem to be clearly differentiated. (3) The accessions classified asH. compressum present biochemical phenotypes quite different from the rest of the species. (4)H. stenostachys, H. muticum andH. chilense constitute a group of variable species with many biochemical similarities and close phylogenetic relationships. (5) The evolutionary pattern of these American species seems to follow a model of reticulate evolution.     

Relationships between three species ofFestuca sect.Bovinae (Poaceae)

Seed protein electrophoresis confirms the existence of polymorphism among hexaploid populations ofFestuca arundinacea. Both protein and morphological results suggest thatF. pratensis andF. arundinacea should retain independent specific status. High protein homology of these two species withF. gigantea points towards phylogenetic links between these taxa.     

Tests for the presence of gametoclonal variation in barley and wheat doubled haploids produced using the Hordeum bulbosum system

Summary To investigate whether the Hordeum bulbosum system of doubled haploid production generates gametoclonal variation, populations of second generation doubled haploid lines were developed from first generation doubled haploid lines of two barley varieties and three wheat genotypes. In barley, no variation between doubled haploids from doubled haploids was detected for a range of quantitative characters, suggesting the absence of any gametoclonal effects. However, the original selfed-seed stocks were shown to contain cryptic allelic variation for some of the characters investigated. In wheat, gametoclonal variation was detected for ear emergence time, plant height and yield, and its components for two out of the three genotypes investigated. The type and range of variation was similar to that reported from studies of somaclonal variation from immature embryos and gametoclonal variation from anther culture. Generally, the effects appeared to reduce the yield performance of individual lines. The difference in response between the two species and the consequences for the use of the doubled haploid system in breeding programmes are discussed.     

Mapping of Rym14 Hb,a gene introgressed from Hordeum bulbosum and conferring resistance to BaMMV and BaYMV in barley

Hordeum bulbosum represents the secondary gene pool of barley and constitutes a potential source of various disease resistances in barley breeding. Interspecific crosses of H. vulgare × H. bulbosum resulted in recombinant diploid-barley progeny with immunity to BaMMV after mechanical inoculation. Tests on fields contaminated with different viruses demonstrated that resistance was effective against all European viruses of the soil-borne virus complex (BaMMV, BaYMV-1, -2). Genetic analysis revealed that resistance was dominantly inherited. Marker analysis in a F5 mapping family was performed to map the introgression in the barley genome and to estimate its size after several rounds of recombination. RFLP anchor-marker alleles indicative of an H. bulbosum introgression were found to cover an interval 2.9 cM in length on chromosome 6HS. The soil-borne virus resistance locus harboured by this introgressed segment was designated Rym14^Hb. For marker-assisted selection of Rym14^Hb carriers, a diagnostic codominant STS marker was derived from an AFLP fragment amplified from leaf cDNA of homozygous-resistant genotypes inoculated with BaMMV.Communicated by F. Salamini     

Gene lineage analysis in populations ofHelianthus niveus andH. petiolaris (Asteraceae)

Helianthus petiolaris andH. niveus are polytypic species which are morphologically distinct at the periphery of their ranges but intergrade in areas of sympatry.Helianthus niveus includes both annual and perennial members, whereasH. petiolaris is strictly annual. Chloroplast DNA and nuclear ribosomal DNA restriction site data were used to reconstruct the evolutionary history of populations of the two species. Cladistic analyses reveal the following: (1) neither species is monophyletic; (2) the annual habit is derived once in this complex; and (3) the region of morphological intergradation appears to be primary in origin. The significance of interbreeding versus common descent in defining species concepts is discussed in relation to the above cladistic analyses.     

Development of embryo and endosperm inEragrostis unioloides (Poaceae)

The embryo development conforms to the Asterad type or Megarchetype II. A single cotyledon called scutellum is derived from the entire tierl. The shoot apex, lateral in position, and the coleoptile, originate from the tierl. The remaining parts of the mature embryo are derived from the tiers situated belowl. The short suspensor takes its origin from the lowermost tierp. The endospermab initio is free nuclear in development. It becomes cellular first around the proembryo. Physiologically, the ripe endosperm represents a dead tissue.     

Chloroplast DNA study of the generaAmbrosia s.l. andHymenoclea (Asteraceae): Systematic implications

The relationship ofAmbrosia (ragweed) andFranseria has long been debated. Their treatment as separate genera has been repeatedly challenged. In this study, chloroplast DNA restriction site variation was examined for species from bothAmbrosia andFranseria as well as taxa from the closely related genusHymenoclea. The chloroplast genomes of members of these three genera were examined using 21 restriction endonucleases and the restriction mutations were used to construct phylogenetic trees. Wagner and Dollo parsimony as well as weighted parsimony were employed to compare the different phylogenies. The results support a close relationship betweenAmbrosia andFranseria, but indicate that the two groups are well separated. Compared toFranseria, Ambrosia is a much more strongly supported group, and the results indicate thatHymenoclea is closer toFranseria than toAmbrosia. The cpDNA phylogeny was used as a framework to examine evolutionary trends in morphology and secondary chemistry.