Chromosome elimination in Hordeum vulgare x H. bulbosum hybrids

Summary The hybrid progeny from a stable amphidiploid of H. vulgare x H. bulbosum involving the cultivar Vada and an unstable amphidiploid involving the cultivar Emir were studied. The genotypes examined contained two genomes from Vada or one from Vada and one from Emir, with one or two genomes from H. bulbosum. Comparisons between the chromosome numbers in root-tips and anthers revealed that there was no chromosome elimination in most plants, whether there was one or two Vada genomes present. The one plant in which chromosome elimination was positively identified had Emir as opposed to Vada cytoplasm. It also had a high incidence of degraded or fragmented chromosomes in the PMCs. Differences in stability between a 27 chromosome plant and other hypotetraploids suggest that Vada contains both elimination genes and elimination suppressor genes. Upon selfing, again irrespective of the number of Vada genomes present, circa triploid hybrids gave rise to diploid H. vulgare offspring while hypotetraploids produced hybrid-like plants. These included diploids, triploids and tetraploids. There was evidence that suggested that H. vulgare as well as H. bulbosum chromosomes had been eliminated.     

Variable patterns of chromosome synapsis at pachytene in Hordeum vulgare x H. bulbosum hybrids and their parents

Synaptonemal complexes (SC) have been analysed in barley (Hordeum vulgare), H. bulbosum and two F, hybrids between them. These hybrids show different recombination frequencies and at pachytene show significant differences in the total length of SC formed and in the extent of synapsis. Higher recombination frequency in the hybrids was correlated with a greater total SC length. Differences in SC length were also observed between the parental species with H. bulbosum having a greater SC length than H. vulgare. However, species and hybrid can have similar SC lengths but clearly different recombination frequencies and, therefore, the relationship between SC length and recombination is not clear-cut.     

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.     

Karyotypes,nucleoli, and amphiplasty in hybrids between Hordeum vulgare L. and H. bulbosum L.

Chromosome measurements were carried out in Hordeum vulgare, H. bulbosum, and their diploid, triploid, and tetraploid hybrids. The chromosomes were classified by using relative values, and thus karyotypes were established. For comparison of these karyotypes both relative and absolute values were used. It was concluded that differential amphiplasty occurred, whereas neutral amphiplasty could not be demonstrated. In the hybrids the relative length of the parts of the chromosomes (long arm, short arm, satellite) was not changed in comparison with these lengths in the pure species. The karyotypes of both species had considerable similarities. From comparing the mean absolute genome lengths, it was, however, concluded that in the pure species, as well as in all hybrid types, the chromosomes of H. vulgare were longer than those of H. bulbosum. In the diploid and tetraploid hybrids the mean genome lengths were shorter than those in the pure species and the triploid hybrids. The differential amphiplasty was such that the secondary constriction of chromosome 6 of H. bulbosum, did not show up in the hybrids. This could be related to the suppression of nucleolar formation in the genome of H. bulbosum, because the maximum number of nucleoli in root tip cells equalled the number of satellite chromosomes. Finally it was found that the pattern of nucleolar fusion in diploid and triploid hybrids deviated from the expectation. The results were discussed in relation to chromosomal disturbances that occurred in the hybrid tissues and that resulted in elimination of chromosomes and other effects.     

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.     

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.     

Morphology and cytology of Hordeum chilense X H. bulbosum hybrids

Summary An interspecific hybrid between Hordeum chilense and H. bulbosum was produced. The hybrid resembles the male parent, but some characters from H. chilense are also present. Transgressive inheritance for other characters has also been observed. Neither chromosome instability nor homoeologous pairing was found.     

Genome relationships between Hordeum vulgare L. and H. bulbosum L.

Interrelationships between H. vulgare (2x=14) and H. bulbosum (2x=14; 4x=28) were estimated on the basis of the karyotypes and the pairing behaviour of the chromosomes in diploid, triploid and tetraploid hybrids obtained with the aid of embryo culture. — A comparison of the karyotypes of the two species revealed similarities as well as differences. It was concluded that at least 4 or more of the chromosomes were similar in morphology and probably closely related. — Diploid and tetraploid hybrids are rarely obtained and their chromosome numbers tend to be unstable whereas triploid hybrids (1 vulgare + 2 bulbosum genomes) were stable and relatively easy to produce. In the diploid hybrid only 40% of the meiotic cells contained 14 chromosomes while the numbers ranged from 7 to 16 in other cells. All hybrids exhibited pairing between the chromosomes of the two species. Diploid hybrids had a mean of 5.0 and a maximum of 7 bivalents per cell in those cells having 14 chromosomes. Triploid hybrids from crosses between 2x H. vulgare and 4x H. bulbosum exhibited a mean of 1.5 and a maximum of 5 trivalents per cell. In a hexaploid sector found following colchicine treatment of a triploid the mean frequencies of chromosome associations per cell were: 5.5_I+8.0_II+0.7_III+3.7_IV+0.3_V+0.4_VI. One unstable 27 chromosome hybrid obtained from crosses between the autotetraploid forms had a mean of 1.1 and a maximum of 4 quadrivalents per cell. The chromosome associations observed in these hybrids are consistent and are taken as evidence of homoeologous pairing between the chromosomes of the two species. Interspecific hybridization between these two species also reveals that chromosome stable hybrids are only obtained when the genomes are present in a ratio of 1 vulgare2 bulbosum. Based upon the results obtained, the possibility of transferring genetic characters from H. bulbosum into cultivated barley is discussed.     

The mildew resistance of Hordeum bulbosum and its transference into H. vulgare genotypes

Mildew from susceptible genotypes (SI and S2) of Hordeum bulbosum was found to be avirulent on all H. vulgare genotypes tested, including such cultivars as Proctor with no known genes for resistance to mildew. The H. bulbosum genotype SI (2n =14) proved resistant to all isolates of mildew from H. vulgare. The mildew isolates used for these tests possessed most of the common virulence factors which attack the current ‘vulgare’ cultivars in Western Europe. Some H. bulbosum genotypes were resistant to the ‘bulbosum’ mildew. Attempts at combining resistance from both species are presented and discussed.     

Elimination of chromosomes in Hordeum vulgare x H. bulbosum crosses at mitosis and interphase involves micronucleus formation and progressive heterochromatinization

Uniparental chromosome elimination occurs in several interspecific hybrids of plants. We studied the mechanism underlying selective elimination of the paternal chromosomes during the development of Hordeum vulgare x H. bulbosum hybrid embryos that is restricted to an early stage of development. In almost all embryos most of the H. bulbosum chromatin undergoes a fast rate of elimination within nine days after pollination. There are differences in the mitotic behaviour between the parental chromosomes, with H. bulbosum chromatids segregating asymmetrically at anaphase. We provide evidence for a chromosome elimination pathway that involves the formation of nuclear extrusions during interphase in addition to postmitotically formed micronuclei. The chromatin structure of nuclei and micronuclei differs and heterochromatinization and disintegration of the nuclear envelope of micronuclei are the final steps of chromosome elimination.     

Plant regeneration and variants from calli derived from immature embryos of diploid barley (Hordeum vulgare L.) and H. vulgare L. x H. bulbosum L. crosses

Summary Plant regeneration from calli was carried out at two locations using several parental genotypes and environments. Selfed immature diploid (VV) barley (Hordeum vulgare) embryos and immature haploid (V) or hybrid (VB) embryos from crosses between H. vulgare and H. bulbosum were used as explants. Golden Promise, Emir and CB 7432 were the best cultivars for plant regeneration, and 15°C tended to be more suitable for plant development than higher temperatures. A total of 844 regenerants were obtained, and over 200 fertile progenies were screened agronomically. Apart from the occurrence of polyploidy and albinism, three variants were identified. One showed maternal inheritance for yellow leaf striping while the other two were controlled by single recessive genes. One of these possessed increased resistance to scald (Rhynchosporium secalis) compared with the donor parent cv Triumph, and one resembled a chlorina mutant.     

PCR detection of Hordeum bulbosum introgressions in an H. vulgare background using a retrotransposon-like sequence

Retrotransposon-like sequences are ideal tools for initial screening assays to distinguish between closely related species because of their ubiquitous presence, high copy number, chromosome coverage and rapid sequence evolution. A retrotransposon-like sequence, pSc119.1, cloned from Secale cereale (rye) has been used to obtain PCR primers that are capable of detecting small introgressions of Hordeum bulbosum (bulbous barley grass) chromatin in a Hordeum vulgare (cultivated barley) background. Combining this PCR-based assay with a crude but effective high-throughput DNA extraction has enabled the rapid identification of plants possessing H. bulbosum introgressions from large numbers of progeny from H. vulgare×H. bulbosum crosses. These plants are then further characterized by more-refined cytological, molecular and pathological techniques to locate and map the introgressed chromatin and to evaluate their disease resistance. Received: 18 April 2001 / Accepted: 23 August 2001     

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.     

New hybrids of Hordeum parodii with Hordeum vulgare,H. bogdanii,Agropyron caninum and X Triticosecale

Summary Hybrids were produced from crossing Hordeum vulgare, H. bogdanii, Agropyron caninum and × Triticosecale onto H. parodii (6x or 4x). The rates at which hybrids were produced, expressed in terms of plantlet establishment as percent pollinated florets, ranged from 0.47%, (6x H.parodii × 6x × Triticosecale cv. Welsh) to 6.3% (4x H. parodii × 2x H. vulgare cv. Betzes). Based on frequencies of paired configurations at MI, autosyndetic pairing appeared to be promoted by the presence of a Secale cereale genome but suppressed by the genome of H. vulgare.Contribution No. 759 Ottawa Research Station     

Specific Isozyme Marker of a Virus Resistant Barley Derived from Interspecies Cross Between Hordeum vulgare and H. bulbosum

A diploid barley cultivar "Supi 1" was crossed with a tetraploid Hordeum bulbosum “GBC141” to transfer the disease resistant traits. Eleven viable triploid F1 plants were produced by means of embryo rescue technique. The resulting triploid hybrids were backcrossed to diploid barley, and seven BC1 plants were obtained. One of the BC1 plants exhibited barley yellow mosaic virus (BaYMV) resistance when grown in the diseased nursery. Isozyme analysis of H. vulgate, H. bulbosum and their backcross hybrids were made via slab polyacrylamide gel electrophoresis technique. The primary results showed that zymogram variation could be obviously found between diploid barley "Supi 1' and tetraploid H. bulbosurn "GBC141”. A peroxidase isozyme (Rf=0.47) from H. bulbosum was detected in the peroxidase isozyme zymogram of young roots of backcross hybrid BC1-2. This peroxidase isozyme was related to the BaYMV resistance but the linkage relation will be determined by the genetic analysis of the F2 population in the future. The BaYMV resistant line of the backcross with isozyme marker is the important resource of barley disease-resistant breeding.     

Characterization of Hordeum vulgare - Hordeum bulbosum chromosome substitution lines by restriction fragment length polymorphism analysis.

Plants obtained from crosses between Hordeum vulgare and H. bulbosum were previously analyzed cytologically and for isozyme composition. They were identified as possessing substitutions of one or more H. vulgare chromosomes by their H. bulbosum homoeologues. To confirm their constitution and assess the merits of molecular techniques, chromosome-specific probes developed for the Triticeae were hybridized to Southern blots of DNA extracted from these plants and their parents. The hybridization patterns in the substitution plants confirmed that particular chromosomes of H. vulgare were replaced by their H. bulbosum homoeologues. For most probes, heterozygosity between pairs of H. bulbosum chromosomes was recorded. A possible duplication involving H. bulbosum homoeologues of barley chromosomes 4 and 7 was observed. Although molecular and cytological methods for analyzing chromosomally engineered plants are complementary, molecular probes may uncover differences not discernible using light microscopy or isozyme analysis.     

Characterization by RFLP analysis and genomic in situ hybridization of a recombinant and a monosomic substitution plant derived from Hordeum vulgare L. x Hordeum bulbosum L. crosses.

Interspecific crosses in Hordeum have been made with the aim of transferring desirable traits, such as disease resistance, from a wild species, Hordeum bulbosum, into cultivated barley (Hordeum vulgare). Interspecific recombinants have previously been identified using several methods, but there are limitations with all the techniques. We improved our ability to characterize progeny from H. vulgare x H. bulbosum crosses by using genomic in situ hybridization (GISH). The plant material comprised a recombinant and a monosomic alien substitution plant derived from H. vulgare x H. bulbosum crosses. The recombinant possesses a pubescent leaf sheath conferred by a gene transferred from H. bulbosum into barley cultivar Golden Promise. The use of GISH on a plant homozygous for the pubescence gene confirmed the presence of H. bulbosum DNA located distally on two barley chromosomes and we mapped the introgression to barley chromosome 4HL using RFLP analysis. Furthermore, by means of an allelism test we found that the transferred gene for pubescence is allelic or closely linked to a gene for pubescence (Hs) located on barley chromosome 4HL. The presence of a single H. bulbosum chromosome in the monosomic substitution plant was confirmed by GISH. A distal introgression of H. bulbosum DNA was also observed on one barley chromosome, which was located on chromosome 3HL by RFLP analysis.     

Cytogenetic diversity of SSR motifs within and between Hordeum species carrying the H genome: H. vulgare L. and H. bulbosum L.

Non-denaturing FISH (ND-FISH) was used to compare the distribution of four simple sequence repeats (SSRs)—(AG) _n , (AAG) _n , (ACT) _n and (ATC) _n —in somatic root tip metaphase spreads of 12 barley (H. vulgare ssp. vulgare) cultivars, seven lines of their wild progenitor H. vulgare ssp. spontaneum, and four lines of their close relative H. bulbosum, to determine whether the range of molecular diversity shown by these highly polymorphic sequences is reflected at the chromosome level. In both, the cultivated and wild barleys, clusters of AG and ATC repeats were invariant. In contrast, clusters of AAG and ACT showed polymorphism. Karyotypes were prepared after the identification of their seven pairs of homologous chromosomes. Variation between these homologues was only observed in one wild accession that showed the segregation of a reciprocal translocation involving chromosomes 5H and 7H. The two subspecies of H. vulgare analysed were no different in terms of their SSRs. Only AAG repeats were found clustered strongly on the chromosomes of all lines of H. bulbosum examined. Wide variation was seen between homologous chromosomes within and across these lines. These results are the first to provide insight into the cytogenetic diversity of SSRs in barley and its closest relatives. Differences in the abundance and distribution of each SSR analysed, between H. vulgare and H. bulbosum, suggest that these species do not share the same H genome, and support the idea that these species are not very closely related. Southern blotting experiments revealed the complex organization of these SSRs, supporting the findings made with ND-FISH.     

Intergeneric hybrids between Triticum crassum and Hordeum vulgare

Summary Intergeneric hybrids between Triticum crassum (2n=6x=42) and Hordeum vulgare cv. Bomi were obtained at a frequency of 15% of pollinated florets. Meiotic chromosome pairing in the hybrids was not different from that observed in a polyhaploid of T. crassum indicating negligible pairing between chromosomes of the two species and secondly that the genome of H. vulgare had no effect on intergenomic pairing in T. crassum.Contribution No. 646 Ottawa Research Station     

Bulb Histogenesis in Hordeum bulbosum L.

The histological and associated histochemical phenomena occurringin the developing bud of Hordeum bulbosum were investigated.Naturally the bulb is formed by swelling of the lowest internodeof the spike-bearing stern. Bulb initiation began by a rapidcell division followed by cell elongation. The bulb form wasachieved by forming cell files directed diagonally outward inthe upper part and by a periphery meristern contributing cellsto the middle and the lower parts of the bulb internode. Starchaccumulated in the cells before bulb initiation and was utilizedcompletely only in the elongating cells when bulb elongationoccurred. High content of nucleic acids and total protein, demonstratedby specific stainings, was observed in the cytoplasm of cellsof the intercalary meristem at each internode. In the bulb,these stainings became progressively less as the bulb elongated.