A maternal factor conferring resistance to crown rust in Lolium multiflorum cv.'Axis'

The genetic basis for crown rust resistance was analysed in the progeny of reciprocal crosses between Lolium multiflorum plants that were either resistant (cv. ‘Axis’) or susceptible to crown rust. The progeny harvested from the resistant parent was two‐ to seven‐fold more resistant than the progeny harvested from the susceptible parent. These data indicate the presence of a maternal factor which increases resistance to crown rust in some of the ryegrass plants used as parents.     

Introgression of crown rust resistance from Festuca spp. into Lolium multiflorum

Crown rust, Puccinia coronata Corda, causes one of the most damaging foliar diseases in Italian ryegrass, Lolium multiflorum Lam. For introgression of crown rust resistance, highly resistant hybrids of the crosses Festuca arundinacea (2n = 6x = 42) ×L. multiflorum (2n = 4x = 28) and reciprocally, L. multiflorum (2n = 2x = 14) ×Festuca pratensis (2n = 4x = 28) and subsequently resistant recombinant individuals were used as female parents and susceptible cultivars of Italian ryegrass as male parents in three successive backcrosses. The BC₃ plants were selfed and crossed mutually. Uredospores of seven different crown rust isolates collected from plants of L. multiflorum, Lolium perenne, F. pratensis, F. arundinacea and L. multiflorum × F. pratensis hybrids were applied to identify the resistance or susceptibility of the parental species, backcrossed, selfed and intercrossed progenies. The various crown rust populations revealed a species-specific capability to infect plants of the Lolium-Festuca complex corresponding to the host species from which the spores originated. Selected BC⁹ plants, however, were found to be completely resistant to all crown rust populations tested. Successful introgression of the resistance was achieved from F. arundinacea as well as from F. pratensis. The resistance represents a dominant character, apparently based on a strong heterologous incompatibility between host and pathogen. In phenotype, bivalent formation during meiosis and in fertility, the novel germplasms are comparable with the L. multiflorum cultivars.     

Infection of Lolium and Festuca spp. by Drechslera siccans and D. Catenaria

Summary Drechslera siccans was the commonest of the species of this leaf spot pathogen found on single spaced plants of Lolium perenne at Aberystwyth. D. catenaria and D. dictyoides were also frequently isolated. A wide range of genotypes within the host species L. perenne, L. multiflorum, Festuca arundinacea and F. pratensis, and structural hybrids between F. arundinacea and L. multiflorum, were all equally susceptible to conidial infection by D. siccans in the glasshouse and showed similar development of leaf spot symptoms, even though Festuca spp. are not normally attacked in the field. However, when exposed to infection out of doors there was significant variation in resistance, both between species and often also between genotypes within the same species. As in so many other instances of fungal infection, tetraploidy appeared to confer a higher level of resistance. The degree of resistance of a F. arundinacea × L. multiflorum amphiploid and two succeeding backcross generations to L. multiflorum approximated to that of the more susceptible L. multiflorum parent. By contrast to D. siccans, when plants were inoculated with D. catenaria in the glasshouse, both intervarietal and intravarietal variation in leaf spot development was evident. Resistance to D. siccans appears to depend partly on a limitation of conidial production, and to D. catenaria on restriction of hyphal growth following infection, which may explain the relative discrepancies between field and glasshouse results. The implications for resistance breeding are discussed.     

Androgenesis from a Lolium multiflorum × Festuca arundinacea hybrid to generate extreme variation for freezing-tolerance

New opportunities for plant breeding using androgenesis in Lolium × Festuca hybrids have been identified. Plants derived by anther culture from a Lolium multiflorum × Festuca arundinacea (5x) hybrid were screened for freezing-tolerance, and their post-freezing recovery compared. The androgenic population showed extreme diversity in freezing-tolerance. While the majority of androgenic plants had inferior freezing-tolerance compared with the freezing-sensitive L. multiflorum parent, 6% of the population were more freezing resistant than the freezing-tolerant Festuca parent. Novel Lolium and Festuca gene combinations resulting from rare meiotic events were recovered within the androgenic population. The two most freezing-tolerant androgenic plants carried virtually the entire F. pratensis subgenome of F. arundinacea. F. pratensis is known to carry genes for freezing-tolerance and would be expected to be the primary source of genes governing this trait within the F. arundinacea genome. The most freezing-tolerant androgenic plants were more freezing-tolerant than the hybrid plant from which they were derived. Consequently, androgenesis was also effective in removing factor(s) reducing the expression of freezing-tolerance within the L. multiflorum × F. arundinacea (5x) hybrid.     

Cytogenetic Studies of Hybrids between Festuca gigantea Vill. and Lolium multiflorum Lam.

The meiotic behaviour of the hybrid between Festuca gigantea (2n – 6x = 42) and Festuca gigantea (2n = 6x = 42) indicates distinct structural differences between the two species. These differences are also apparent in the degree of chromosome pairing observed in Lolium multiflorum×F. gigantea compared with previous reports on the L. multiflorum×F. arundinacea hybrids. Although the L. multiflorum×F. gigantea 8x amphiploid com Dines the complementary characters of the two species and is agronomically interesting, there are some irregularities in meiotic behaviour that could affect the stability of the amphiploid. Seed fertility in the amphiploid is high and preliminary studies show evidence of a high degree of self-fertility. The pentaploid hybrid between autotetraploid L. multiflorum×F. gigantea is sufficiently fertile to be used as the pollen plant in crosses with diploid L. multiflorum (BC₁). Backcrossing the BC₃ hybrid to L. multiflorum results in mainly diploid progeny. The possibilities, of using this crossing scheme to introgress F. gigantea characters into L. multiflorum is discussed as an alternative approach to amphiploid breeding as a means o: combining specific complementary characters of the two species.     

Androgenesis in a Lolium multiflorum × Festuca arundinacea hybrid to generate genotypic variation for drought resistance

The pentaploid hybrid of Lolium multiflorum and Festuca arundinacea (2n = 5x = 35) combines the high growth rate of L. multiforum with the drought resistance and freezing-tolerance of F. arundinacea. Unfortunately, it also displays the deleterious traits associated with Festuca, namely those associated with high leaf fibre content giving rise to poor palatability and digestibility. To access different combinations of these characters, anther cultures were initiated and regenerated into single embryo derived plants. The anther culture method was very productive since out of a total of 2349 androgenic plants derived from the same parent plant, 57% were green plantlets, although only 507 (22%) subsequently established into plants following transfer to soil. Chromosome counts of randomly selected lines showed that plants with euploid chromosome numbers (14, 21, and 28) would appear to have selective advantage during regeneration. There was wide variation between mature androgenic plants grown under field conditions in plant height, leaf length, leaf width, tiller number and herbage dry matter. The variation between genotypes in response to drought stress was assessed by placing replicate clones under rain-out shelters or under irrigated control conditions in the field. Herbage dry matter under drought was higher in a number of androgenic lines than either parents, but not higher than the pentaploid hybrid. Androgenesis was shown to be a highly effective procedure to expose diverse phenotypic variation all derived from the same Lolium × Festuca hybrid genotype. This revised version was published online in July 2006 with corrections to the Cover Date.     

Habitat and management affect genetic structure of Festuca pratensis but not Lolium multiflorum ecotype populations

Genetic diversity present in permanent grassland may be valuable for broadening gene pools in breeding programmes and for conservation of genetic resources. However, little is known about the amount of genetic diversity present at specific habitats and about site‐related factors affecting it. To identify valuable habitats, genetic diversity of 12 ecotype populations and four reference cultivars of both Festuca pratensis Huds. and Lolium multiflorum Lam. was analysed using Simple sequence repeat markers (SSR). Analysis of molecular variance revealed a larger within population variation for L. multiflorum (97.1%) than for F. pratensis (92.6%). F. pratensis ecotype populations were clearly separated from cultivars and formed three distinct subclusters according to the geographic regions they were sampled from. Differences between L. multiflorum ecotype populations and cultivars were small and no grouping of populations was observed. Thus, only F. pratensis ecotype populations were structured and habitat as well as management had a slight influence on genetic structure. This information may allow the design of individual strategies for targeted utilization of genetic resources in plant breeding programmes.     

The controlled introgression of Festuca arundinacea genes into Lolium multiflorum

Summary Using phosphoglucoisomerase (PGI/2) as a genetic marker, it has been shown to be possible to transfer genes from Festuca arundinacea into diploid Lolium multiflorum using the pentaploid hybrid L. multiflorum (4x) x F. arundinacea (6x). The pentaploid hybrid was sufficiently fertile to be used in reciprocal crosses with diploid. L. multiflorum. When used as the male parent, only two backcross generations were then required to reconstitute the diploid genotype. Intergeneric recombinants including a F. arundinacea PGI/2 allele were found among the diploid BC₂. Cytological data indicates that although the majority of chromosome associations involve only homologous Lolium chromosomes, associations involving Lolium and Festuca chromosomes also occur.Interpollinating the pentaploid hybrids prior to commencing a backcrossing programme increases the number of cycles of recombination and improves the chance of recovering intergeneric recombinants. The crossing programme described is proposed to be an effective method of introducing F. arundinacea genes into L. multiflorum.     

First in vivo assessment of feeding value of Festulolium hybrids derived from Festuca arundinacea var. glaucescens and selection for palatability

This paper reports on the first results of feeding value experiments, assessed with sheep, of hybrids between tetraploid Italian ryegrass (Lolium multiflorum, 2n = 4×= 28) and a progenitor of tall fescue, Festuca arundinacea var. glaucescens (2n = 4×= 28). Three hybrid populations resulting from either high, low or no selection for palatability were compared with a variety of Italian ryegrass as control and two varieties of the cultivated hexaploid (2n = 6×= 42) tall fescue, one of which had been selected for high palatability. On average, the hybrid populations were found to be as palatable as the improved tall fescue with voluntary intake (VI), in vivo digestibility of organic matter (DOM), and net energy expressed in fodder units for milk, similar to the best palatable tall fescue and to Italian ryegrass. However, selection for improved palatability among hybrids did not result in any further improvement of other traits, except VI in the hybrid selected for high palatability which significantly exceeded VI of Italian ryegrass. In conclusion, intergeneric hybridization between L. multiflorum and F. arundinacea var. glaucescens led to an immediate increase of feeding value compared with selection for palatability in tall fescue. However, to improve feeding value of hybrids further, similar selection for high palatability appeared questionable and possibly detrimental for other agronomic traits because of complicated inheritance in tetraploid L. multiflorum×F. glaucescens hybrids.     

Disturbed Mendelian segregations at isozyme marker loci in early backcrosses of Lolium multiflorum × Festuca pratensis hybrids to L. multiflorum

Summary The segregation of interspecific recombinant Festuca pratensis (Fp) alleles, introgressed into the germolasm of Lolium multiflorum (Lm), at four loci (PGI/2, AcP/2, GOT/3 and BAP) is described. Heterozygous (Lm/Fp) plants were backcrossed to L. multiflorum (2n=2×=14) and subsequent BC2 Lm/Fp sibling genotypes intercrossed.In crosses between BC1 heterozygous plants (Lm/Fp) used as males and L. multiflorum, there was a reduced transmission of the F. pratensis (Donor Parent) derived alleles in the populations with PGI/2 and AcP/2 marker loci compared to the reciprocal cross but the reduction was not significantly different in those with GOT/3 and BAP markers.Two PGI/2 marked BC2 half-sib families in particular exhibited a more extreme deficiency of Fp/Fp progeny plants than anticipated from the BC2 segregations indicating possible linkage to zygotic lethals. Deficiencies of F. pratensis alleles were, in most cases, less marked in BC2 half-sib families indicating that a further round of recombination had reduced the size of the introgressed chromosome segment or that deleterious linkages had been broken. A tendency towards heterozygote advantage was found in one BAP marked halfsib family.The significance for forage grass breeding of reduced transmission rates of Donor Parent alleles in early back-cross generations especially through the male gametes is discussed.     

Genomic constitution of Festulolium cultivars released in the Czech Republic

Genomic in situ hybridization (GISH) was used to characterize the chromosome constitutions of individual plants from a set of tetraploid and hexaploid cultivars of Festulolium developed and released in the Czech Republic from hybrids of Lolium multiflorum with Festuca pratensis and F. arundinacea. A simplified GISH protocol readily discriminated parental genomes in the hybrids and facilitated the screening of large numbers of plants per accession. The contribution of parental genomes in the cultivars tested ranged from predominance of chromatin from one of the parents to a more balanced contribution from both parents. However, in none of the cultivars were equal proportions of chromatin from both parents present. The parental contribution to the hybrids was both in the form of complete chromosomes or as chromosome translocations. In hexaploid cultivars from (L. multiflorum × F. arundinacea) × F. arundinacea hybrids the average numbers of complete L. multiflorum chromosomes ranged from 4.95 to 7.5 and the numbers of translocations from 6.33 to 10.21. Two tetraploid cultivars from (L. multiflorum × F. arundinacea) × L. multiflorum hybrids showed a strong prevalence of L. multiflorum chromatin and intergeneric translocations were rare. In the tetraploid cultivar ‘Perun’ of the L. multiflorum × F. pratensis hybrid there were 11.7 chromosomes of L. multiflorum and 14.7 recombined chromosomes on average. Reasons for the domination of one of the parental genomes in hybrid cultivars are not clear and are only partially explained by breeding history. Recombination rates of individual genomes in hybrids involving F. arundinacea were evaluated in double hybridization experiments. The results indicated a strong affinity of the L. multiflorum genome for the F. pratensis genome present in F. arundinacea and little affinity for the F. glaucescens genome. This suggests that introgressions from F. arundinacea into L. multiflorum are primarily limited to the F. pratensis genome which can be more readily accessed in L. multiflorum × F. pratensis hybrids.     

Inheritance of resistance to Puccinia coronata Corda and Rhynchosporium orthosporum Caldwell in Italian ryegrass

Summary Progeny analysis within Italian ryegrass (Lolium multiflorum) cv. RvP infected with a single clone of crown rust (Puccinia coronata f. sp. lolii) revealed major factors, one probably a single dominant gene, conferring resistance in two of twelve randomly selected parents. The remaining variation in resistance to this pathogen, and all of the variation in resistance to Rhynchosporium orthosporum, was controlled by a relatively large number of minor genes exhibiting neither dominance nor epistasis. There was no significant maternal inheritance. Absence of any correlation between progeny scores for reaction to the two pathogens indicated that resistance to each was inherited independently.     

Value of permanent grassland habitats as reservoirs of Festuca pratensis Huds. and Lolium multiflorum Lam. populations for breeding and conservation

European natural and semi-natural grassland form reservoirs of genetic resources containing highly adapted and variable ecotype populations of forage plants. Variation within these reservoirs is stimulated by variation in natural and anthropogenic site-related factors. Changes in agricultural practices lead to the loss of many characteristic habitats. In order to preserve resources for breeding, targeted conservation strategies for germplasm in gene banks (ex-situ) or on site (in-situ) are needed. In order to define site-related criteria for the potential of habitats to preserve valuable resources for breeding and conservation, 38 different habitats across Switzerland were selected to collect Festuca pratensis Huds. and Lolium multiflorum Lam. ecotype populations. Phenotypic variation and population differentiation of 60 single plants were evaluated in a field experiment using 16 morpho-physiological traits. For F. pratensis, ecotype populations and cultivars were clearly separated and there was a significant correlation between diversity of morphological traits and geographic location of sampling sites. For L. multiflorum no clear separation of ecotype populations and cultivars was observed suggesting gene flow from adjacent temporary leys into permanent grassland. Several ecotype populations were superior to cultivars in important traits such as early heading or resistance to winter damage, indicating the importance of natural habitats as a reservoir of genetic resources for breeding. In conclusion, maintenance of permanent grassland in contrasting environments appears to be a promising strategy for preserving valuable genetic variation of forage grasses. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Verification of the introgression of Erianthus arundinaceus germplasm into sugarcane using molecular markers

Erianthus arundinaceus has a number of important agronomic traits including good ratooning ability, tolerance to both drought and water logging, disease resistance and vigour and is of interest as a potential source of parental germplasm to sugarcane breeders. However, to date, attempts to produce fertile hybrids between sugarcane (Saccharum spp.) and E. arundinaceus have been unsuccessful. Microsatellite markers that generated genus‐specific markers and were highly polymorphic within sugarcane were identified. The microsatellite markers and 5S rDNA PCR were used to screen intergeneric (F1) clones from Saccharum officinarum×E. arundinaceus crosses, and two Saccharum backcross progeny (BC1) populations derived from crosses between selected F1 clones and sugarcane (Saccharum spp.), to identify genuine Saccharum spp. BC1 progeny. The 5S rDNA PCR marker and highly polymorphic microsatellites with Erianthus‐specific bands confirmed the F1 parent of the two putative BC1 populations was a S. officinarum×E. arundinaceus hybrid and allowed the identification of the genuine BC1 progeny from selfs of the F1 parent. This is the first verification of BC1 progeny from an F1 intergeneric hybrid x sugarcane (Saccharum spp.) clone with molecular markers and confirms the introgression of E. arundinaceus germplasm into sugarcane. It should now be possible to exploit genes of value from E. arundinaceus in sugarcane breeding programmes.     

Identification of a molecular marker linked to an Agropyron elongatum-derived gene Lr19 for leaf rust resistance in wheat

The leaf rust resistance gene Lr19, transferred from Agropyron elongatum into wheat (Triticum aestivum L.) imparts resistance to all pathotypes of leaf rust (Puccinia recondita f.sp. tritici) in South‐east Asia. A segregating F₂ population from a cross between the leaf rust resistant parent ‘HW 2046’ carrying Lr19 and a susceptible parent ‘Agra Local’ was screened in the phytotron against a virulent pathotype 77‐5 of leaf rust with the objective of identifying the molecular markers linked to Lr19. The gene was first tagged with a randomly amplified polymorphic DNA (RAPD) marker S73₇₂₈. The RAPD marker linked to the gene Lr19 which mapped at 6.4 ± 0.035 cM distance, was converted to a sequence characterized amplified region (SCAR) marker. The SCAR marker (SCS73₇₁₉) was specific to Lr19 and was not amplified in the near‐isogenic lines (NILs) carrying other equally effective alien genes Lr9, Lr28 and Lr32 enabling breeders to pyramid Lr19 with these genes.     

Inheritance of resistance to anthracnose caused by Colletotrichum capsici in Capsicum

Inheritance of resistance to anthracnose caused by Colletotrichum capsici (Syd.) Butler & Bisby was studied in interspecific Capsicum populations derived from a cross between a Thai elite cultivar Capsicum annuum L.‘Bangchang’ and a resistant line C. chinense Jacq.‘PBC932′. The resistance was assessed by measuring lesion area per fruit area (LFA) on detached chili fruits, using a laboratory‐based injection inoculation. Nil symptoms resembling the resistant parent ‘PBC932’ were also identified in the progeny F₂ and BC1 populations. Segregation of resistance (nil LFA) and susceptibility in the F₂ fitted a 1: 3 Mendelian ratio, indicating that resistance was responsible by a single recessive gene. The segregation of the trait in the testcrosses in both BC1s also confirmed the 1: 3 gene segregating model as found in the F₂.     

Androgenesis as a means of dissecting complex genetic and physiological controls: selecting useful gene combinations for breeding freezing tolerant grasses

The advantages of androgenesis from Lolium × Festuca hybrids as a means towards enhanced gene expression and capture of rare genetic variation are reviewed. New evidence is presented for the technique’s use in combination with introgression-mapping for targeting Festuca-derived genes for enhanced freezing-tolerance. As a starting point, a dihaploid genotype derived by androgenesis from a Lolium multiflorum × Festuca pratensis amphiploid (2n = 4x = 28) hybrid cultivar is used as female parent in a backcross breeding programme with L. multiflorum (2n = 2x = 14). A derivative of the backcross␣breeding programme was a genotype of L.␣multiflorum (2n = 2x = 14) incorporating a F.␣pratensis introgression on chromosome 4 that was more freezing-tolerant than Lolium. New evidence of the importance in Lolium and Festuca species of the adaptive capabilities of Photosystem II (PSII) in relation to subsequent freezing-tolerance, is presented. Non-photochemical quenching (NPQ) mechanisms for expulsion of excess light energy during cold acclimation are found in F. pratensis but not in L. multiflorum. Screens of a backcross population derived from an initial dihaploid genotype (n + n = 14) produced by androgenesis from a L. multiflorum × F.␣pratensis amphiploid, indicate a direct relationship between cold acclimation induced increases in NPQ and freezing-tolerance. Preliminary evidence of a role for genes found on chromosome 4 of F. pratensis for increased NPQ expression, is presented.     

Genetic analysis of forage grasses based on heterologous RFLP markers detected by rice cDNAs

Genetic polymorphism within and between three species of forage grasses, perennial ryegrass (Lolium perenne L), meadow fescue (Festuca pratensis Huds.) and tall fescue (Festuca arundinacea Schreb.), was analyzed using restriction fragment length polymorphism (RFLP) markers detected by rice cDNA probes developed at the Rice Genome Research Programme of Japan (RGP). One hundred and ninety‐seven rice cDNA clones were used for hybridization to genomic DNA of forage grasses. Many of the rice cDNA clones produced no visible band or only a smear with no discrete bands. Twenty‐three clones showed high efficiency cross‐hybridization to the genomic DNA of forage grasses. Genetic variation was evaluated for five varieties and one population of forage grasses using 12 polymorphic rice cDNA RFLP probes. Genetic variability within varieties as measured by Rogers’ genetic distance was considerably lower for the F. pratensis variety ‘Tomosakae’ than for the L. perenne and F. arundinacea varieties. To determine the genetic diversity between varieties of different species, cluster analysis was performed using data from the 12 RFLP probes. The two accessions of Lolium perenne were clustered more closely together than the three varieties of F. arundinacea. Two Japanese varieties of F. arundinacea were grouped in the same cluster. The variety‐specific RFLP markers were seen among six accessions of L. perenne, F. pratensis and F. arundinacea. Such variety‐specific RFLP markers would provide very useful tools for breeding programmes such as the intergeneric hybridization of Lolium and Festuca genera.     

Breeding for hard red winter wheat cultivars adapted to conventional-till and no-till systems in northern latitudes

Summary Monosomic alien addition lines combining individual F. drymeja chromosomes and the L. multiflorum complement were isolated from the cross between the triploid hybrid L. multiflorum (4x) × F. drymeja (2x) and diploid L. multiflorum (2x). Chromosome pairing in the addition lines was studied at metaphase 1 of meiosis and the relationship between single F. drymeja chromosomes and the corresponding homologous pair in L. multiflorum is discussed. Trivalent frequency in the addition lines was higher than expected from observations of chromosome pairing in the triploid hybrid and there were differences between lines in the number of trivalent associations formed. There is some evidence to suggest that trivalent frequency is not entirely dependent on chromosome length and that transmission of the alien chromosome in the female is dependent on the size of the added chromosome. Morphological studies were made to assess the phenotypic effects of the addition of single F. drymeja chromosomes to the L. multiflorum complement. Two plants (2n=14) with recombination between a L. multiflorum and a F. drymeja chromosome were identified.     

Resistance to Pseudomonas solanacearum in the potato: I. Effects of sources of resistance and adaptation

Summary F1 hybrid true potato seed progenies with multiple sources of specific resistance to Pseudomonas solanacearum and adaptation were evaluated under tropical greenhouse conditions for resistance to a race 1 isolate of P. solanacearum. Results indicated that genes for adaptation are involved in conferring resistance to bacterial wilt. The effect of a particular resistant parent clone on the resistance level of its progeny depended on the resistance, adaptation or the combination of both characteristics of its mate. A heat tolerant parent gave a higher frequency of resistant offspring in combination with an ascertained source of resistance. Combining ability was an apparent feature of resistance to bacterial wilt and resistance was highest in the majority of the crosses that had a wide genetic background for both resistance and adaptation. The possible genetic nature of the resistance to bacterial wilt is discussed.