RESUMEN
Genes for winter hardiness and frost tolerance were introgressed from Festuca arundinacea into winter-sensitive Lolium multiflorum. Two partly fertile, pentaploid (2n = 5x = 35) F(1) hybrids F. arundinacea (2n = 6x = 42) x L. multiflorum (2n = 4x = 28) were generated and backcrossed twice onto L. multiflorum (2x). The backcross 1 (BC(1)) and backcross 2 (BC(2)) plants were preselected for high vigor and good fertility, and subsequently, a total of 83 BC(2) plants were selected for winter hardiness after 2 Polish winters and by simulated freezing tests. Genomic in situ hybridization (GISH) was performed on 6 winter-hardy plants selected after the first winter and shown to be significantly (P < 0.05) more frost tolerant than the L. multiflorum control. Among the analyzed BC(2) winter survivors, only diploid (2n = 2x = 14) plants were found. Five plants carried 13 intact L. multiflorum chromosomes and 1 L. multiflorum chromosome with a single introgressed F. arundinacea terminal chromosome segment. The sixth BC(2) winter survivor appeared to be Lolium without any Festuca introgression capable of detection by GISH. A combined GISH and fluorescence in situ hybridization analysis with rDNA probes of the most winter-hardy (after 2 winters) and frost-tolerant BC(2) plant revealed the location of an F. arundinacea introgression on the nonsatellite arm of L. multiflorum chromosome 2, the same chromosome location reported previously as a site for frost tolerance genes in the diploid and winter-hardy species Festuca pratensis.
Asunto(s)
Adaptación Biológica/genética , Mapeo Cromosómico , Festuca/genética , Técnicas de Transferencia de Gen , Lolium/genética , Estaciones del Año , Cruzamiento/métodos , Genotipo , Hibridación Fluorescente in Situ , Plantas Modificadas GenéticamenteRESUMEN
The first backcross breeding programme for the transfer of freezing-tolerance genes from winter hardy Festuca pratensis to winter-sensitive Lolium multiflorum is described. A partly fertile, triploid F(1) hybrid F. pratensis (2n=2x=14) x L. multiflorum (2n=4x=28) was employed initially, and after two backcrosses to L. multiflorum (2x) a total of 242 backcross two (BC(2)) plants were generated. Genomic in situ hybridisation (GISH) was performed on 61 BC(2) plants selected for their good growth and winter survival characters in the spring following one Polish winter (2000-2001). Among the winter survivors, diploid chromosome numbers were present in 80% of plants. An appropriate single Festuca introgression in an otherwise undisturbed Lolium genome could provide increased freezing tolerance without compromise to the good growth and plant vigour found in Lolium. Among all the diploids, a total of 20 individuals were identified, each with a single F. pratensis chromosome segment. Another diploid plant contained 13 Lolium chromosomes and a large metacentric F. pratensis chromosome, identified as chromosome 4, with two large distal Lolium introgressions on each chromosome arm. Three of the diploid BC(2), including the genotype with Festuca chromosome 4 DNA sequences, were found to have freezing tolerance in excess of that of L. multiflorum, and in one case in excess of the F. pratensis used as control. A detailed cytological analysis combining GISH and fluorescence in situ hybridisation analyses with rDNA probes revealed that the other two freezing-tolerant genotypes carried a Festuca chromosome segment at the same terminal location on the non-satellite arm of Lolium chromosome 2.
Asunto(s)
Aclimatación/genética , Cruzamiento , Festuca/genética , Congelación , Lolium/genética , Plantas Modificadas Genéticamente/genética , Cruzamiento/métodos , Cruzamientos Genéticos , Festuca/fisiología , Técnicas de Transferencia de Gen , Genoma , Hibridación in Situ , Hibridación Fluorescente in Situ , Lolium/fisiología , Plantas Modificadas Genéticamente/fisiologíaRESUMEN
Androgenesis using amphidiploid cultivars of Festuca pratensis x Lolium multiflorum as parents, overcame earlier problems that gave rise to widespread plant sterility amongst androgenic Festulolium populations. Two Festuca pratensis x Lolium multiflorum (2n = 4x = 28) cultivars, Sulino and Felopa, were highly amenable to androgenesis and 10% of plants, including some novel androgenic genotypes, had sufficient fertility to produce progeny and further generations. The genomes of amphidiploid cultivars, which represent the F8 generation, were the result of considerable intergeneric chromosome recombination. Moreover, during cultivar development, natural and breeders' selection pressures had led to the assembly of gene combinations that conferred good growth characters and fertility with the removal of putative deleterious gene combinations. Over 80% of the androgenic plants derived from the amphidiploid F. pratensis x L. multiflorum (2n = 4x = 28) had 14 chromosomes and were likely to be dihaploids with a single genome of Lolium and of Festuca. In contrast, hybrids of F. pratensis x L. multiflorum (2n = 2x = 14) found naturally are invariably sterile. Structural reorganization within the genomes of the androgenic Festulolium plants had restored fertility in genotypes expected to contain the haploid genome of Lolium and Festuca. This provided opportunities for their future incorporation in breeding programmes and the development of fertile diploid Lolium-Festuca hybrids. Amongst the androgenic plants, Festulolium genotypes were recovered that conferred excellent drought resistance or freezing tolerance and were thought to be highly suitable for entry into plant breeding programmes.
Asunto(s)
Genes de Plantas/genética , Poaceae/genética , Polen/fisiología , Adaptación Fisiológica/efectos de los fármacos , Adaptación Fisiológica/genética , Cruzamiento , Frío , Diploidia , Genotipo , Hibridación Genética , Lolium/genética , Poaceae/efectos de los fármacos , Poaceae/fisiología , Poliploidía , Reproducción/genética , Selección Genética , Agua/farmacologíaRESUMEN
Diploid and tetraploid forms of Lolium multiflorum and Festuca pratensis were crossed under controlled conditions and after embryo rescue all four combinations of autoallotriploid hybrids were obtained. Male and female fertility and chromosome pairing at metaphase I of meiosis were studied in several plants from each hybrid combination. The hybrids with two genomes of L. multiflorum and one of F. pratensis (genomic formulae LmLmFp and FpLmLm) were male and female fertile while the hybrids with two genomes of F. pratensis and one of L. multiflorum had a reduced fertility (FpFpLm) or were completely sterile (LmFpFp). Chromosome pairing at metaphase I varied among hybrid combinations depending on their genomic composition. LmLmFp and FpLmLm hybrids had similar patterns of pairing (1.83I + 5.29II + 2.85III and 2.22I + 5.22II + 2.75III, respectively) but they differed from those of FpFpLm (3.65I + 4.65II + 2.68III) and especially from LmFpFp (4.78I + 5.87II + 1.49III). Conventional analysis of meiosis failed to explain the differences in chromosome behaviour and fertility/sterility levels between the autoallotriploid hybrids with two Lolium or two Festuca genomes.