Colchicine-induced heritable variation in cell size and chloroplast number in the leaf cells of inbred ryegrasses (Lolium perenne,L. multiflorum)

Summary A 3 hr period of treatment with a 0.2% aqueous solution of colchicine was given to one week old seedlings of 3 inbred lines of perennial ryegrass (Lolium perenne) and 4 lines of Italian ryegrass (L. multiflorum). The surviving plants developed as mixoploids and were self-pollinated. In the following generation (CT1) the seeds were grown without further treatment, and were screened cytologically at the seedling stage to separate out diploids from tetraploids. Only the diploids were kept, and in this CT1 generation comparisons were then made between controls (2x) and colchicine-treated diploids (C2x) for leaf mesophyll cell size and chloroplast number variation. Heritable differences were found such that the cell plan areas and/or the numbers of chloroplasts per cell were significantly greater in the C2x compared with the 2x treatment in 3 and 5 out of the 7 lines. In each case there was enhancement of chloroplast numbers independently of the variation in cell size.     

Components of resistance to ryegrass mosaic virus in a clone of Lolium perenne and their strain-specificity

Summary The extreme resistance to ryegrass mosaic virus (RMV) of a clone of Lolium perenne L. was due to a combination of two distinct types of resistance: resistance to infection and resistance to multiplication and movement of virus within the plant. Resistance to infection was quantitatively inherited and highly effective against three strains of RMV, while resistance to multiplication and movement was controlled by two complementary recessive genes and was effective against only two of the three RMV strains.     

Genetics of two mechanisms of resistance to Meloidogyne naasi (FRANKLIN) in an Aegilops variabilis Eig. accession

Summary Accession No. 1 of Aegilops variabilis has complete resistance to the root knot nematode, Meloidogyne naasi. F2 segregation in a cross between this and two susceptible accessions showed that one dominant gene named Rkn-mn1 prevented development of galls on the roots and consequently of female nematodes. The study of the numbers of females in galls on the F2 plants allowed detection of a recessive gene, Rkn-mnAv, suppressing development of J2 larvae into females. The presence of Rkn-mnAv also resulted in a decrease of the level of galling. Rkn-mn1 has already been introduced into wheat. The interest in transferring also Rkn-mnAv is discussed in relation to extending durability of the nematode resistance.     

Introduction of a self-compatible gene of Lolium temulentum L. to perennial ryegrass (Lolium perenne L.) for the purpose of the production of inbred lines of perennial ryegrass

Interspecific hybrids between self-compatible species, Lolium temulentum L. and self-incompatible species, L. perenne L. were obtained using embryo rescue. Two cycles of backcrossing of interspecific hybrids with L. perenne were carried out. A 1: 1 segregation ratio of self comatibility and incompatibility was observed in backcross generations. These segregation data confirmed that self-compatibility of L. temulentum was controlled by a single gene. It suggested that the self-compatible gene of L. temulentum could be introduced to self-incompatible plants of L. perenne through interspecific hybridization. It appears that utilization of a self-compatible gene of L. temulentum would be useful for production of inbred lines of L. perenne. The possibility of breeding procedures of perennial ryegrass using a self-compatible gene was discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Specialization of crown rust (Puccinia coronata Corda) on clones of Italian ryegrass (Lolium multiflorum Lam.)

Summary Cultures of crown rust (Puccinia coronata Corda) from a mixed uredospore population obtained from plants in 16 localities in England and Wales were multiplied on four of 10 initially resistant clones of Italian ryegrass (Lolium multiflorum Lam.). Only two of the four cultures were found to be most pathogenic on the clones on which they had been multiplied. One clone became very susceptible to its own culture but the other became only moderately susceptible. Clone/culture interactions occurred which were not obviously related to the clone of origin of the culture. Thus, there was considerable genetic diversity in the crop which would tend to limit the multiplication of new races. There was a strong correlation between visual score for susceptiblity and uredospore production, showing the value of visual scores for resistance breeding.     

Variation in digestibility in nine populations of perennial ryegrass (Lolium perenne) and its genetic basis

Summary Thirty five individuals from each of nine perennial ryegrass populations were sampled from set-stocked and zero-grazed swards and from an old permanent pasture. The swards were 6, 18, 30 months and 11 years old when sampled. The permanent pasture was at least 40 years old.Dry matter digestibility (DMD) and organic matter digestibility (OMD) were determined for green leaf material harvested 4 days after ear emergence. Significant differences were found between the populations, but genotypic differences were found only for OMD. The largest proportion of the total variation was due to differences between blocks. In the zero-grazed populations, DMD and OMD declined significantly with increasing sward age. This was not found for the set-stocked swards. Two sets of pair crosses, from which mid-parent, progeny regressions could be determined, were carried out.The regressions of offspring on mid-parent for set-stocked and zero-grazed swards were used to assess the presence of additive genetic variation and to estimate narrow sense heritability of DMD. The coefficients of correlation, 0.122 and 0.226 respectively, were non-significant, indicating an absence of additive genetic variation.     

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.     

Intergeneric symmetric and asymmetric somatic hybridization in Festuca and Lolium

Summary Intergeneric symmetric and asymmetric somatic hybrids have been obtained by fusion of metabolically inactivated protoplasts from embryogenic suspension cultures of tall fescue (Festuca arundinacea Schreb.) and unirradiated or 10–500 Gy-irradiated protoplasts from non-morphogenic cell suspensions of Italian ryegrass (Lolium multiflorum Lam.). Genotypically and phenotypically different somatic hybrid Festulolium mature flowering plants were regenerated.Species-specific sequences from F. arundinacea and L. multiflorum being dispersed and evenly-represented in the corresponding genomes were isolated and used for the molecular characterization of the nuclear make-up of the intergeneric, somatic Festulolium plants recovered. The irradiation of Italian ryegrass protoplasts with 250 Gy X-rays prior to fusogenic treatment favoured the unidirectional elimination of most or part of the donor chromosomes. Irradiation of L. multiflorum protoplasts with 500 Gy produced highly asymmetric (over 80% donor genome elimination) nuclear hybrids and clones showing a complete loss of donor chromosomes.The RFLP analysis of the organellar composition in symmetric and asymmetric tall fescue (+) Italian ryegrass regenerants confirmed their somatic hybrid character and revealed a bias towards recipient-type organelles when extensive donor nuclear genome elimination had occurred.Approaches aimed at improving persistence of ryegrasses based on asymmetric somatic hybridization with largely sexually-incompatible grass species (F. rubra and Alopecurus pratensis), and at transferring the cytoplasmic male sterility trait by intra- and inter-specific hybridization in L. multiflorum and L. perenne, have been undertaken.Abbreviations cpDNA chloroplast DNA - CMS cytoplasmic male sterility - 2,4-D 2,4-dichlorophenoxy-acetic acid - IOA iodoacetamide - mtDNA mitochondrial DNA - RFLP restriction fragment length polymorphism     

Heritability and gain from selection for field resistance against multiple root-knot nematode species (Meloidogyne incognita race 1 and M. javanica) in carrot

Heritability and gain from selection of traits associated with field resistance to multiple root-knot nematode species were estimated in carrot (Daucus carota L.). Experimental plots were uniformly and simultaneously infested with Meloidogyne incognita race 1 and M. javanica. Forty-seven half-sib families derived from the cv. ‘Brasília’ were evaluated for: (1) percentage of commercial roots with non-galling symptoms (%RNG); (2) percentage of commercial roots with gall symptoms (%RGS) and (3) percentage of non-commercial roots (%NCR). The cultivars ‘Brasília’ (resistant), ‘Carandaí’ (intermediate) and ‘Nova Kuroda’ (susceptible) were employed as standards. Broad-sense heritability estimates were 61.9 % for %RNG; 30.6 % for %RGS and 67.9 % for %NCR. However, the gains from selection were very small for all traits. The genotypic correlation between %RGS and %NCR was –0.38 and the correlation between %RNG and %RGS was 0.13. Selection on increasing %RNG resulted in a negative correlated response on %NCR (genotypic correlation = –0.99) indicating repulsion phase linkage(s) between the genetic factor(s) controlling these traits. The phenotypic correlation of –0.88 between %RNG and %NCR suggested that incomplete penetrance and dosage effects might be involved in the expression of these traits. Selection of superior genotypes (i.e. high %RNG, low %RGS, and low %NCR) can be achieved. ‘Brasília’ was among the genotypes selected for the following cycle of recombination reinforcing the notion that this cultivar is one of the most promising sources of stable, wide-spectrum field resistance to Meloidogyne species in D. carota. This revised version was published online in July 2006 with corrections to the Cover Date.     

A statistical mixture model for estimating the proportion of unreduced pollen grains in perennial ryegrass (Lolium perenne L.) via the size of pollen grains

Summary The size of pollen grains is commonly used to indicate the ploidy level of pollen grains. In this paper observations of the diameter of pollen grains are evaluated from one diploid accession of perennial ryegrass (Lolium perenne L.), which was expected to produce diploid (unreduced) pollen grains in addition to haploid pollen grains. The considerable overlap of the diameter distributions of haploid and diploid pollen grains severely hampers the accurate estimation of the proportion of diploid pollen grains. To overcome this problem we develop in this paper a statistical normal mixture model and we describe a method to test for the production of diploid pollen grains from a diploid parent, and to estimate the proportion of diploid pollen grains.     

Seed yield related to crop development and to yield components in nine cultivars of perennial ryegrass (Lolium perenne L.)

Summary We had previously found differences for seed yield among nine perennial ryegrass cultivars which were not associated with variation for seed weight. To detect the physiological basis of these genetic differences for seed yield, growth analyses were carried out. We related crop development and components of seed yield to seed yield during three years on clay and sandy soil. No significant differences occurred among cultivars for accumulation and partitioning of dry matter or the pattern of tiller production. Seed yield of the cultivars was not associated with ear number or total dry matter yield of the seed crop. Seed yield was more correlated with the number of seeds per unit area than with seed weight. The number of seeds as calculated after harvest from seed yield and seed weight was much lower than the number of seeds as estimated prior to harvest from seed yield components. The number of spikelets differed significantly among the cultivars, but the ranking was different from that for seed yield. The physiological basis of the genetic differences for seed yield is not clear. Implications for breeding perennial ryegrass are discussed.     

Barley yellow dwarf virus in relation to the breeding and assessment of herbage grasses for yield and uniformity

Summary Ryegrass plants (Lolium spp.) rarely developed obvious foliar symptoms when infected with barley yellow dwarf virus, but their yield was often substantially reduced. Perhaps of even greater importance, the virus modified their morphological development and seasonal growth rhythm. Consequently, the disease introduces considerable non-heritable variation into populations of grass plants so its possible presence may introduce difficulties into selecting varietal mother plants for specific growth characteristics and when assessing bred varieties for uniformity.Virus infection caused the greatest deviation in growth morphology and the smallest reduction in yield in early-flowering hay type varieties: the converse was true of late-flowering pasture type varieties. Unless a source of immunity or extreme resistance to the virus can be found therefore, it may be difficult to combine minimum deviation in growth morphology and minimum yield loss in a single variety.     

Development of a PCR-based selection assay for root-knot nematode resistance ( Rmc1) by a comparative analysis of the Solanum bulbocastanum and S. tuberosum genome

A PCR-based assay has been developed for marker assisted selection of root-knot nematode resistance ( Rmc1) in potato. To this end, a comparative genome analysis was carried out between Solanum bulbocastanum and S. tuberosum to identify PCR-based chromosome 11 alleles linked to Rmc1. The use of co-migrating AFLP markers, obtained by using primer combinations previously applied for AFLP analysis of the S. tuberosum genome, failed to align the AFLP map of the S. bulbocastanum genome with the S. tuberosum map. Apparently, the S. bulbocastanum genome is genetically too distantly related to the S. tuberosum genome for this type of analysis. Cleaved amplified polymorphic sequence (CAPS) markers were more readily applied for a comparative analysis within the region of interest. Rmc1 could be localized within a 4 cM interval between markers CT182 and M39b. It is demonstrated that the resistance spectrum of Rmc1 includes not only Meloidogyne chitwoodi and the related species M. fallax but also a genetically distinct population of M. hapla. The cost-efficiency of the CAPS markers applied for Rmc1 renders this approach as an attractive alternative for screening large segregating populations of potato for root-knot nematode resistance. This revised version was published online in July 2006 with corrections to the Cover Date.     

Possibilities of family selection in a cultivar of lucerne,Medicago sativa L., for resistance to the stem nematode,Ditylenchus dipsaci (Kühn) Filipjev

Summary Half-sib families representing the lucerne cultivar Resis were tested for resistance to a stem nematode population, recovered from lucerne. The percentage resistance of the HS-families varied from 13 to 88%. The mean resistance for the cultivar was approximately 50%. Half-sib family selection would be more efficient than recurrent mass selection. The observed data suggest the action of a few genes with relatively large effects.     

The production of alien monosomic additions in Beta vulgaris as a source for the introgression of resistance to beet root nematode (Heterodera schachtii) from Beta species of the section Patellares

Summary Experiments were carried out for adding the chromosome carrying resistance to beet root nematode (Heterodera schachtii) from the wild Beta species of the section Patellares (B. procumbens, B. webbiana and B. patellaris) to the genome of B. vulgaris. Preliminary experiments indicated that crosses between the wild species and B. vulgaris cultivars of the mangold type yielded on average more viable F₁ hybrids than crosses with sugar and fodderbeet. However, crossability varied strongly between individual parental combinations. It was concluded that most types of B. vulgaris can be hybridized with the wild species of the section Patellares if a sufficient number of pair-crosses is made. Crosses between diploid cultivars or species of the section Vulgares and diploid wild species of the section Patellares yielded many hybrids which, however, were highly sterile. From crosses between tetraploid B. vulgaris and the wild species a great number of viable allotriploid and allotetraploid hybrids was obtained. In the backcross progenies of allotriploid hybrids 26% alien monosomic additions occurred, of which 4.1% carried the resistance bearing chromosome of B. procumbens or B. patellaris. The programme will be continued by sereening progenies of the resistant monosomic addition plants for the occurrence of resistant disomic introgression products.