Inheritance and chromosomal location of Septoria passerinii resistance introgressed from Hordeum bulbosum into Hordeum vulgare

Septoria speckled leaf blotch (SSLB), caused by Septoria passerinii, has become one of the most serious diseases of barley in the Upper Midwest region of the USA. The recombinant line 36L5 derived from a backcross of the susceptible barley cultivar ‘Emir’ and a resistant Hordeum bulbosum parent Cb2920/4/Colch was found to be resistant to S. passerinii. Two doubled haploids derived from 36L5 were backcrossed to cv.‘Emir’ to obtain two BCF₂ populations for determining the inheritance of resistance to S. passerinii. BCF₂ progeny and BCF_2:3 families were evaluated at the seedling stage in the greenhouse for reaction to S. passerinii. BCF₂ progeny and BCF_2:3 families from both crosses segregated 3 : 1 (resistant : susceptible), and 1:2:1 (resistant : segregating : susceptible), respectively, indicating that the H. bulbosum‐derived SSLB resistance is conferred by a single dominant gene. The H. bulbosum introgressions were positioned on chromosome 4HL by genomic and fluorescent in situ hybridizations (GISH and FISH, respectively) and by Southern hybridization with the rye repetitive sequence pSc119.2. These findings indicate that SSLB resistance in H. bulbosum has the potential to be transferred and utilized in barley breeding programs.     

The evaluation of superior Hordeum bulbosum L. genotypes for use in a doubled haploid barley breeding programme

Summary Eight Hordeum bulbosum selections were produced from a cross between Cb 2920/4 and Cb 2929/1, two genotypes widely used in doubled haploid breeding programmes. The selections were hybridized with barley to evaluate their ability to produce high proportions of well-differentiated haploid embryos compared with Cb 2929/1 as control. We report here an initial small-scale investigation followed by a larger-scale test in two different environments to assess seed setting, haploid embryo differentiation rates and VB hybrid formation. These VB embryos contain both parental sets of chromosomes and occur more frequently in the glasshouse during the winter. Two of the eight selections were identified as combining the desirable characteristics of both parents, namely high seed setting on cv. Vada which is partially incompatible with H. bulbosum, large numbers of well-differentiated haploid embryos and a low incidence of VB hybrids. The selections are available for release to interested research workers and plant breedersAbbreviations DH doubled haploid - VB a hybrid from H. vulgare x H. bulbosum which contains both parental sets of chromosomes     

Association of leaf rust and powdery mildew resistance in a recombinant derived from a Hordeum vulgare × Hordeum bulbosum hybrid

While studying powdery mildew resistance in a recombinant line (code 81882) derived from a Hordeum vulgare (cv. ‘Vada’) ×Hordeum bulbosum hybrid, a low infection type of resistance to leaf rust was observed. To determine the mode of inheritance of the leaf rust resistance and whether there was linkage between the two resistances, F₂ and F₃ progenies from crosses between 81882 and ‘Vada’ were inoculated with the leaf rust and powdery mildew pathogens. Southern blots were prepared using restricted DNA extracted from leaves of 82 F₂ plants and four chromosome 2HS sequences were hybridized with the blots to define the length of the introgression. The leaf rust resistance appears to be inherited as a single dominant gene on chromosome 2HS, which co-segregates with the powdery mildew resistance. There was an almost complete association between the resistances and the respective molecular markers, but it is likely that the strong linkage results from the frequent inheritance of the introgressed H. bulbosum DNA as an intact segment of chromatin with only low levels of recombination within the segment.     

The resistance of Hordeum bulbosum and its hybrids with H. vulgare to common fungal pathogens

Summary Two tetraploid and two diploid clones of Hordeum bulbosum were screened for resistance to five isolates of powdery mildew which are virulent on cultivated barley. All were resistant and this resistance was also expressed in hybrids with H. vulgare. The tetraploid genotypes were also resistant to isolates of yellow rust and brown rust. These results show that H. bulbosum contains useful genes for resistance to these diseases and that there is a potential to transfer these into cultivated barley.     

Comparison of Anther Culture and Hordeum bulbosum Method for the Production of Doubled Haploid in Winter Barley I. Production of Green Plants

The relative response to two-major techniques of producing haploids in barley – anther culture and the Hordeum bulbosyum method – were compared using seven F₁ hybrids from a winter barley breeding programme. Both techniques were Applied to the same plants with the same number of replications. 358 viable green plants were obtained from 420 spikes ot donor plants used: 147 through anther culture and 211 through the H. bulbosum method. From the data analysis we found that the response to the two techniques was not significantly different, genotype effect and genotype × technique interaction were highly significant. The possibility of combining the two techniques in a doubled haploid production programme is discussed.     

Anther Culture-Derived Regenerants from Hordeum vulgare × H. bulbosum Crosses

Hordeum bulbosum has several desirable attributes, including disease resistance, which would be worthwhile transferring to H. vulgare. Despite homoeologous chromosome pairing in the interspecific hybrids, there have been few reports of successful gene introgression between the two species. A possible explanation for this is that recombinant male gametes are at a competitive disadvantage with normal balanced gametes during post-pollination events. To circumvent this problem, the possibility of obtaining plants directly from immature pollen grains was investigated. Anthers from diploid, triploid and tetraploid H. vulgare × H. bulbosum hybrids were cultured on defined media. Only hybrids with dehiscent anthers in vivo responded in culture, and after transfer of calli and embryoids to regeneration medium, 36 albino and 12 green plants were obtained. Seven of the green regenerants survived, one of which contained 15 H. vulgare chromosomes (including one acrocentric chromosome) and one H. bulbosum chromosome. Another regenerant (Ac166) resembled a diploid H. vulgare × H. bulbosum hybrid but had partial anther dehiscence and a slightly modified chromosome constitution. Mostly normal H. vulgare progeny were obtained from crosses between H. vulgare cv.‘Emir’ and Ac166, but three plants involved chromosome additions and substitutions.     

Genotype-environment interaction of barley doubled haploids with regard to malting quality

The genotype-environment (GE) interaction of barley doubled haploid (DH) lines was studied in relation to some malting quality characters. DH lines were obtained, using the bulbosum method, from F₁ hybrids between the cultivars ‘Grit’ and ‘Havila’. Thirty DH lines, the parental forms, F₂ and F₃ hybrids of ‘Grit’בHavila’ and the standard cultivar ‘Rudzik’ were studied in a series of field experiments repeated at six environments. Malt protein content, Kolbach index, extract yield and malt fine-coarse difference were measured. The statistical analysis was performed using the computer program SERGEN. For protein content, it was found that most of the lines studied were stable, i.e. their GE interactions were not significant. Among the malt traits, phenotypic values of Kolbach index and fine-coarse difference were most dependent on environmental conditions. For Kolbach index, only one line was found to be stable, while for fine-coarse difference, all lines displayed interaction with the environment.     

An Evaluation of Hordeum vulgare L. ×Psathyrostachys fragilis (Boiss.) Nevski Crosses for Doubled Haploid Barley Production

Investigations were carried out to assess the suitability of the intergeneric cross Hordeum vulgare×Psathyrostachys fragilis for haploid barley production. H. vulgare cvs. ‘Emir’ and ‘Vada’ were each pollinated with P. fragilis P.I. 343192 and plants regenerated from embryos cultured on a modified B5 medium. Seed sets on ‘Vada’ were significantly lower than on ‘Emir’, and all the planes from ‘Vada’×P. fragile remained hybrid. Several of these flowered but there was little pairing between the parental chromosomes. Most of the plants from ‘Emir’×P. fragilis died, as seedlings but 3 plants developed into haploid barley. Because of the practical limitations of pollen availability from P. fragilis and the inconsistencies in haploid plant formation, it is unlikely that the cross will prove as valuable as that between H. vulgare×H. bulbosum for a doubled haploid barley programme.     

The production and behaviour of hexaploid hybrids between Hordeum vulgare and H. Bulbosum

Summary To produce hexaploid (or other polyploid) hybrids, diploid or tetraploid Hordeum vulgare was crossed with hexaploid or octoploid H. bulbosum, and perennial triploid hybrids between the two species were treated with colchicine. The crosses did not yield viable plants: seedset was low, the seed aborted and embryo culture was unsuccessful. The colchicine treatments geve rise to plants in which hexaploid chromosome numbers were observed. At the hexaploid level chromosomal instability occurred, resulting in chromosome elimination.The colchicine-treated triploid hybrids showed in the first years after the treatment better fertility after open flowering than untreated plants, but the level of fertility remained very low. The offspring consisted of haploid, diploid and approximately triploid plants like H. vulgare, tetraploid and approximately tetraploid plants like H. bulbosum, and plants with hybrid morphology and unstable chromosome number, which were highly sterile. Thus the crossing barrier between H. vulgare and H. bulbosum could not be broken down at higher ploidy level.     

Hordeum bulbosum-a new source of disease resistance-transfer of resistance to leaf rust and mosaic viruses from H. bulbosum into winter barley

An outline is given of results for the transfer of new resistances against leaf rust and barley mosaic viruses from Hordeum bulbosum into winter barley. Progenies from backcrosses of barley cultivars with H. bulbosum hybrids were tested both in conventional breeding trials and in additional tests under controlled conditions. Resistance to both pathogens proved to be stable and of good heritability, with differences occurring which depended on the combinations used. Lines with resistance to all leaf rust and mosaic viruses tested, or to either one, were selected. Both resistances segregated independently.     

Genetics of Hordeum chilense– Current Status

Hordeum chilense, a South-American wild barley, is being used in cereal breeding because of its good crossability with both wild and cultivated Triticeae species. Amphiploids and chromosome addition lines of H. chilense in common and durum wheats have been obtained as a means to transfer desirable agronomic characters from wild barley to wheat. No allosyndetic pairing has been detected in diverse hybrids involving H. chilense. Several biochemical markers have been associated with the seven chilense chromosome pairs. New approaches are necessary to transfer genes from H. chilense to cereals.     

Partial incompatibility in crosses between Hordeum vulgare L. and H. bulbosum L.

Summary Certain cultivars of Hordeum vulgare L. gave low seed set (average <30%) after pollination with H. bulbosum, while others consistently gave high seed set (average >80%). The reduced seed setting occurred in crosses with cultivars which had H. distichum var. laevigatum in their ancestry. The results show that the reduced setting is probably controlled by a single dominant gene. This finding is of particular importance to breeders concerned with the use of Vada or some of its high yielding derivatives for producing haploids by interspecific hybridization.     

Gibberellic Acid + 2,4-D Improves Seed Quality in Hordeum vulgare L. ×H. bulbosum L. Crosses

Doubled haploid production from crosses between barley (Hordeum vulgare L.) and H. bulbosum L. is a well established technique for inducing homozygosity. One problem which occurs using certain barley cultivars is premature seed degeneration. In this paper, a post-pollination spray of gibberellic acid and 2,4-dichlorophenoxyacetic acid that delays the onset of senescence is described. The technique improved the number of high quality seeds to more than 75% of the seed set. This improvement should result in increased survival of embryos for culture and subsequent plant regeneration.     

Factors affecting haploid production in wheat using the Hordeum bulbosum system. 2. The effect of the timing of pollination

Summary The effect of manipulating the timing of pollination on the fertilization frequency in wheat × tetraploid Hordeum bulbosum hybridizations was investigated as a possible means of overcoming varietal incompatibility in this cross. Pre- and post-anthesis pollinations were made on previously identified highly crossable and poorly crossable wheat genotypes and the stage of development of the individual florets at pollination was expressed in days from anthesis units.In both crossable and non-crossable wheat genotypes, a significant negative linear relationship was observed between floret age and fertilization frequency. The magnitude of the response of early pollination was dependent on the number of alleles for non-crossability present at the Kr loci. The non-crossable cultivar Highbury, possessing more than one allele for incompatibility, showed a slight response. The greatest response was shown by the highly crossable cultivar Chinese Spring, possessing alleles for crossability at all Kr loci. The single chromosome substitution line, Chinese Spring (Hope 5B), possessing the most potent allele for non-crossability, Kr ₁, showed an intermediate response. The H. bulbosum genotype had no effect on the magnitude of this response, influencing only the mean fertilization frequency.     

Crosses between Hordeum vulgare L. and H. bulbosum L. I. Production,morphology and meiosis of hybrids,haploids and dihaploids

Summary From crosses between diploid and autotetraploid cytotypes of Hordeum vulgare L. (cultivated barley) and H. bulbosum L. (bulbous barley grass) diploid, triploid and tetraploid interspecific hybrids were produced. Both directly and after vegetative segregation crosses in either direction also gave rise to haploids and dihaploids resembling H. vulgare. The use of embryo culture was necessary. Plant morphology of the hybrids was much like that of H. bulbosum, although the hybrid plants were less vigorous. Meiosis in the hybrids was more or less disturbed, and this seemed to be the main cause of the high level of sterility.     

Factors affecting haploid production in wheat using the Hordeum bulbosum system. 3. Post-fertilization effects on embryo survival

Summary An investigation was made of factors affecting the frequency of development of fertilized ovules following wheat x Hordeum bulbosum hybridizations. Seed survival, a term used here to represent the relationship between seed set and fertilization, was shown to vary between the wheat genotypes tested, but was unrelated to the crossability genes present within the wheat genotype. A high seed survival was obtained in crosses involving the variety TH3929 and the single chromosome substitution line, Chinese Spring (Hope 5B). Chinese Spring showed a poor seed survival. Some influence of the H. bulbosum genotype on seed survival was detected. The two environmental factors studied, namely the post-pollination application of gibberellic acid and the ambient temperature, were both shown to affect seed survival. The application of gibberellic acid immediately after pollination or 1 and 2 days after pollination improved seed survival, although the extent of the response was dependent upon the number and timing of the application(s). A genotype-dependent response to the ambient temperature was observed, TH3929 showing a slight but insignificant reduction and Chinese Spring a significant improvement in seed survival with an increase in temperature from 20°C to 26°C. The physiological processes which may be influenced by GA and the ambient temperature and may thereby affect seed survival are discussed.     

The transfer of a gene conferring resistance to scald (Rhynchosporium secalis) from Hordeum bulbosum into H. vulgare chromosome 4HS

Scald is a serious foliar disease that infects barley (Hordeum vulgare L.) causing reduced yields and adversely affecting quality. A means to combat the disease is to breed cultivars that possess genetic resistance. However, all known resistance alleles have so far originated from within the primary genepool of barley. This reliance on H. vulgare and H. vulgare ssp. spontaneum as resistance sources may encourage virulent forms of the pathogen to become established. To broaden the genetic base of cultivated barley and provide novel resistances to many diseases we have used a species from the secondary genepool of barley, H. bulbosum, in a resistance‐breeding programme. In this study we describe the development and trialling of a scald‐resistant recombinant line derived from a hybrid between H. vulgare and H. bulbosum. The scald resistance is simply inherited and located on the short arm of barley chromosome 4 (4HS).     

Attempts to overcome partial incompatibility between Hordeum vulgare L. and H. bulbosum L.

Summary Partial incompatibility has already been reported between Hordeum vulgare and H. bulbosum during the course of doubled haploid production, and in this paper attempts to overcome the breeding barrier are described. The methods which seem to offer most success are those of environmental adjustments and the adoption of new genotypes of H. bulbosum. Further cultivars of H. vulgare exhibiting this phenomenon are also noted.     

Sources of Resistance to Powdery Mildew in Barley Lines Derived from Hordeum spontaneum Collected in Israel

Fourty two barley lines direved from the F₇ of crosses between barley cultivars and different accessions of Hordeum spontaneum collected in Israel and 30 lines or varieties with known genes for resistance to powdery mildew were included m this study. Eleven European and three Israeli powdery mildew cultures, possessing virulence genes corresponding to known resistance genes, were used to make comparisons between the varieties with known resistance genes and H. spontaneum derived lines. The reaction pattern of 39 H. spontaneum derived lines was clearly different from the reaction pattern o; any of the known genes for mildew resistance included in this study. Only two cases were observed in which the reaction pattern of H. spontaneum derived lines agreed with reaction patterns of known genes for mildew resistance viz. Ml-a9 and Ml-p. Trie Mildew resistance of one line apparently traces back to uncontrolled outcrossing with a Ml-a.6+Ml-g resistant cultivar. Since the majority of the 42 host genotypes tested showed distinctive variation in resistant reaction types against different mildew cultures, this study docs not support the assumption that differences in resistant infection types against distinct mildew cultures are sufficient to indicate the presence of supplementary genes for resistance in a given genotype of the host. The results justify the conclusion that the natural population of H. spontaneum in Israel forms a large gene pool for mildew resistance which is not yet used m cultivated barley.     

The effect of pollination bag type on seed quality and size in Hordeum inter- and intraspecific hybridization

Summary The effects of different spike covering materials have been investigated in crosses between Hordeum vulgare and H. bulbosum and in selfed H. vulgare. It was found that after lemmas and paleas were clipped, improvements in seed quality (H. vulgare × H. bulbosum) and weight (selfed H. vulgare) were obtained by covering heads with small brown paper bags compared with other treatments. Possible reasons for these effects are discussed but so far the mechanism has not been clarified. However, light is suggested as playing a major role.