Intergeneric hybrids and an amphiploid between Elymus canadensis and Psathyrostachys juncea

Summary Fifty four hybrid plants between Elymus canadensis and Psathyrostachys juncea were obtained by handpollination and embryo culture. The average cross compatibility between both species was 31.2 percent. One amphiploid plant was induced by colchicine treatment. The hybrid and amphiploid plants resembled P. juncea in appearance but showed a higher plant height and dry matter yield than the parents. The hybrids showed extremely low pollen stainability and were completely sterile. With the exception of one plant (2n=3x+1=22), all hybrid plants were allotriploids (SHN, 2n=3x=21). The amphiploid plant (SSHHNN, 2n=6x=42) showed 58.9% pollen stainability and 11.6% seed fertility.Mean chromosome associations of the hybrids and amphiploid at metaphase I were 0.02IV+0.06III+2.03II+16.91I and 0.07III+18.00II+5.85I, respectively. Lagging chromosomes, chromosome bridges, abnormal cytokinesis, and micronuclei were occasionally observed at the anaphase, telophase, or tetrad stage.     

The transfer of triazine resistance from Brassica napus L. to B. oleracea L. III. First backcross to parental species

Summary Atrazine resistant Brassica napus × B. oleracea F₁ hybrids were backcrossed to both parental species. The backcrosses to B. napus produced seeds in both directions but results were much better when the F₁ hybrid was the pollen parent. Backcrosses to B. oleracea failed completely but BC₁s were rescued by embryo culture both from a tetraploid hybrid (2n = 4x = 37; A₁C₁CC) and sesquidiploid hybrids (2n = 3x = 8; A₁C₁C). Progeny of crosses between the tetraploid hybrid and B. oleracea had between 25 and 28 chromosomes. That of crosses between the sesquidiploid hybrid and B. oleracea had between 21 and 27. A few plants that had chromosome counts outside the expected range may have originated from either diploid parthenogenesis, unreduced gametes or spontaneous chromosome doubling during in vitro culture. Pollen stainability of the BC₁s ranged from 0% to 91.5%. All the BC₁s to B. oleracea were resistant to atrazine.     

Intergeneric Hybrids between Aegilops squarrosa and Secale cereale and their Meiotic Chromosome Behaviour

Three hybrid plants with different combinations of D and R genomes were produced from crosses between Aegilops squarrosa (2×: and 4×) and Secale cereale (2× and 4×) using embryo culture. Production frequencies of mature hybrids, having the genomes DR, DDR and DDRR, were 2.0%, 5.2% and 2.2 %, respectively, of florets pollinated. Amphidiploids were obtained directly from the cross between tetraploid parents. The majority of their morphological features were intermediate between those of the parents but, as with rye, the rachis was tough. Diploid and triploid hybrids were completely seed-sterile, whereas the amphidiploid had an average self-fertility of 4.5 % with a range of 0 to 45 %. The somatic chromosome numbers of diploids, triploids and amphidiploids were 2n = 14, 21 and 28, respectively. At meiotic metaphase I, the mean chromosome associations were 0.01 III + 0.26 II + 13.4 I in diploids, 6.96 II + 7.1 I in triploids, and 10.5 II + 7.9 I in amphidiploids. In diploids, pairing between D and R chromosomes was observed in the form of end-to-end types without chiasmata. Homologous pairing between D chromosomes was predominant in plants having two sets of D genomes. Amphidiploids showed a diploid-like chromosome behaviour. Homoeologous pairing was not observed in either triploids or amphidiploids.     

Cytological and Isozyme Evaluation of Tall Fescue × Italian Ryegrass Hybrids

Development of tall fescue (Festuca arundinacea Schreb., 2n = 6x - 42) × Italian rye-grass (Lolium multiflorum Lam., 2n = 2 ×= 14) hybrids would enhance efforts to improve the quality of tall fescue. Two ‘Kentucky 31’ tall fescue בLemtal Italian ryegrass hybrids were obtained via embryo rescue on MS media containing casein hydrolysate, ascorbic acid and sucrose. Chromosome pairing at metaphase I had an average of more than 12 bivalents per cell. Since Festuca-Lolium pairing can account only for seven of the paired chromosomes, intergenomic as well as interspecific chromosome pairing is indicated. There was no cytoplasmic effect on chromosome pairing. To determine if enzymes could be used as genetic markers for distinguishing hybrids from self-contaminants in crosses, zymograms of PGI, 6-PGD, MDH, GOT and ACPH were obtained from parents and hybrids using starch gel etectrophoresis. PGI, 6-PGD and MDH had fewer bands in the diploid ryegrass, as compared with the hexaploid tall fescue and the tetraploid hybrid.     

Production of 27-, 28-, and 56-chromosome apomictic hybrid derivatives between pearl millet (2n=14) and Pennisetum squamulatum (2n=54)

Summary An interspecific hybridization program designed to transfer gene(s) controlling apomixis from Pennisetum squamulatum Fresen. (2n=6x=54) to induced tetraploid (2n=4x=28) cultivated pearl millet, Pennisetum americanum (L.) Leeke resulted in four offtype plants, two with 27 chromosomes and two with 28 chromosomes. These plants were found among 217 spaced plants established from open-pollinated seed of an apomictic 21-chromosome polyhaploid (2n=21) plant derived from an apomictic interspecific hybrid (2n=41) between tetraploid pearl millet and Pennisetum squamulatum. It appeared that a 21- (or 20-) chromosome unreduced egg from the apomictic polyhaploid united with a 7-chromosome pearl millet (2n=2x=14) gamete to produce a 28- (or 27-) chromosome offspring. Meiotic chromosome behavior was irregular averaging from 3.60 to 4.05 bivalents per microsporocyte in the 27- and 28-chromosome hybrids. The 27- or 28-chromosome hybrids, like the 21-chromosome female parent, shed no pollen, but set from 1.8 to 28 seed per panicle when allowed to outcross with pearl millet. Progeny of the 28-chromosome hybrids were uniform and identical to their respective female parents, indicating that apomixis had been effectively transferred through the egg. In addition, a 56-chromosome plant resulting from chromosome doubling of a 28-chromosome hybrid was identified. Pollen was 68 per cent stainable and the plant averaged 2.3 selfed seeds per panicle. Chromosomes of the 56-chromosome plant paired as bivalents (x=10.67) or associated in multivalents. Three to nine chromosomes remained unpaired at metaphase I. Multiple four-nucleate embryo sacs indicated the 56-chromosome hybrid was an obligate apomict. The production of 27-, 28-, and 56-chromosome hybrid derivatives were the results of interspecific hybridization, haploidization, fertilization of unreduced apomictic eggs, and spontaneous chromosome doubling. These mechanisms resulted in new unique genome combinations between x=7 and x=9 Pennisetum species.     

Production and characterization of intergeneric hybrids between Brassica oleracea and a wild relative Moricandia arvensis

Intergeneric crosses were made between Brassica oleracea and Moricandia arvensis utilizing embryo rescue. Six F₁ hybrid plants were generated in the cross‐combination of B. oleracea × M. arvensis from 64 pods by the placenta‐embryo culture technique, whereas three plants were produced in the reciprocal cross from 40 pods by the ovary culture technique. The hybrid plants were ascertained to be amphihaploid with 2n = 23 chromosomes in mitosis and a meiotic chromosome association of (0–3)II + (17–23)I at metaphase I (M I). In the backcross with B. oleracea, some of these hybrids developed sesquidiploid BC₁ plants with 2n = 32 chromosomes that predominantly exhibited a meiotic configuration of (9II + 14I) in pollen mother cells. The following backcross of BC₂ plants to B. oleracea generated 48 BC₃ progeny with somatic chromosomes from 2n = 19 to 2n = 41. The 2n = 19 plants showed a chromosomal association type of (9II + 1I) and a chromosomal distribution type of (9¹/₂ + 9¹/₂) or (9 + 10) at M I and M II, respectively. These facts might suggest that they were monosomic addition lines (MALs) of B. oleracea carrying a single chromosome of M. arvensis that could offer potential for future genetic and breeding research, together with other novel hybrid progeny developed in this intergeneric hybridization.     

Somatic chromosome elimination and meiotic chromosome pairing in the triple hybrid 6x-(Solanum acaule × S. bulbocastanum) × 2×-S. phureja

Summary Two selected hexaploid F₁ clones from the cross Solanum acaule x S. bulbocastanum were intercrossed and the resulting hybrid plants pollinated with the diploid species S. phureja, in order to obtain tetraploid triple hybrids with the same ploidy level as S. tuberosum cultivars (2n=4x=48).Apart from three trihaploids a large population of triple hybrids was obtained, showing chromosome mosaicism in root tip cells (euploid + hypoploid chromosome numbers) and a uniform, mostly hypoploid chromosome number in the pollen mother cells. It is demonstrated that somatic chromosome elimination in the early stages of development is the most probable cause.From detailed pachytene observations as well as from the chromosome associations observed at metaphase I it is evident, that there is normal pairing between the four genomes in the triple hybrids. Although S. bulbocastanum is a quite distinct species with a rigid crossability barrier with S. phureja, the chromosomes of these two species appear to have a high degree of homology. Especially the formation of quadrivalents involving all twelve groups of four homeologous chromosomes, indicated that the four parental genomes (two from S. acaule, one from S. bulbocastanum and one from S. phureja) are not differentiated to the extent of affecting normal pairing and chiasma formation. These results support the view, that the transfer of valuable characters from S. bulbocastanum to S. tuberosum cultivars is feasible even when these characters are polygenically controlled.     

Meiosis and fertility in interspecific hybrids of Nicotiana tabacum L. ×N. glauca Grah. and their derivatives

Nicotiana glauca is of potential interest to breeders as it carries resistance to black root rot of tobacco. Cytological investigations of sexual interspecific hybrids of N. tabacum T′T′TT (2n = 4x = 48) cv. ‘Wiślica’ × N. glauca GG (2n = 2x = 24) were carried out. The analyses of chromosome association at diakinesis and metaphase I in the PMCs of amphihaploid F₁ T′TG (2n = 3x = 36) revealed low variable pairing with 0–5 bivalents. The sterile amphihaploids F₁ were converted into partial female fertile amphidiploids T′T′TTGG by chromosome doubling. Among 36 mature plants obtained, 15 were found to have chromosome numbers (2n = 6x = 72) and were verified as amphidiploids, 9 had (2n = 6x = 70 or 71) chromosomes while the remaining 12 were haploid. True amphidiploids, in spite of quite high chromosome pairing during meiosis, were very different in pollen fertility, ranging from 0% to 85%. Male fertility disturbances did not correlate with the degree of female fertility upon pollination with N. tabacum. Sesquidiploids T′TG (2n = 5x = 60) obtained from backcrossing the amphidiploids to parental tobacco showed more than 22 bivalents, 10–12 univalents and occasional multivalents that indicated the possibility of interchange events between N. tabacum and N. glauca genomes.     

Morphological and cytogenetic studies on the hybrid between bread wheat and <Emphasis Type="Italic">Psathyrostachys huashanica</Emphasis> Keng ex Kuo

Psathyrostachys huashanica Keng ex Kuo (2n = 2x = 14, NsNs), a source of wheat stripe rust, take-all fungus, and powdery mildew resistance with tolerance to salinity and drought, has been successfully hybridized as the pollen parent to bread wheat without using immature embryo rescuing culture for the first time. All of the CSph2b × P. huashanica hybrid seeds germinate well. Backcross derivatives were successfully obtained. F₁ hybrids were verified as intergeneric hybrids on the basis of morphological observation, cytological and molecular analyses. The results obviously showed the phenotypes of the hybrid plants were intermediate between bread wheat and P. huashanica. Chromosome pairing at MI of PMCs in the F₁ hybrid plants was low, and the meiotic configuration was 26.80 I + 0.60 II (rod). Cytological analysis of the hybrid plants revealed the ineffectiveness of the ph2b gene on chromosome association between the parents. Eight RAPD-specific markers for Ns genome were selected for RAPD analysis, and the results indicated that F₁ hybrids contained the Ns genome of P. huashanica. Furthermore, the significance of the finding for bread wheat improvement was discussed.     

Characterization of morphological variation and cold resistance in interspecific somatic hybrids between potato (Solanum tuberosum L.) and S. brevidens Phil.

Summary Somatic hybrids between Solanum tuberosum L. cv. Gracia (2n=4x=48) and Solanum brevidens Phil. (2n=2x=24) were produced via fusion of mesophyll protoplasts. Selection of the protoplast derived putative hybrid calli was based on their vigorous growth. Additive isozyme patterns and chromosome numbers as well as the expression of parental morphological characters have proved the hybrid origin of the selected regenerants. Extensive chromosome loss during the regeneration process resulted in aneuploid hybrids with high frequency. Genomic instability could not be detected in these plants during the period of vegetative propagation. Regenerants from hybrid tissues exhibited wide morphological variation especially in tuber formation. The detailed morphological analysis based on the use of multivariate method (principal component analysis, PCA) enabled to identify morphological groups among the hybrid clones. The positioning of hybrid clones in the PCA space could not be correlated with chromosome numbers. The genomic ratio represented by the tetraploid and diploid parents influenced the morphology of somatic hybrid population according to the applied analytical system. Two selected hybrid clones have exhibited an intermediate degree of frost tolerance compared to the parents, based on the recovery of plants from lower buds after freezing of potted plants.     

Introduction of allien variation into wheat by gene recombination. I. Crosses between mono V (5B) Triticum aestivum L. and Secale cereale L. and Aegilops columnaris zhuk.

An analysis is made of meiotic pairing of hybrids whether or not deficient in the chromosome V (5B) from crosses between Mono V (5B) T. aestivum var. Chinese Spring and Secale cereale and Aegilops columnaris. Transmission rates of deficiency are not equal for both combinations. Isochromosomes are found in the hybrid plants.5B deficiency allows the pairing and recombination between the chromosomes of wheat and those of the related species. Such deficiency may be a useful wheat breeding tool to introduce alien variation into such cultivated species.Effectiveness of 5B deficiency on meiotic pairing of the hybrids, measured as the increase of the proportion of complement paired, is 36–37% and 49–50% in the hybrids involving rye and Ae. columnaris respectively. Minimum percentages of the introduction of alien variation into wheat are established for the material investigated: 2% for rye and 28% for Ae. columnaris.Methods of utilisation of the 5B deficiency in a programme of wheat breeding are discussed and outlined.     

Cytology and Fertility of Hybrids and Amphiploids between Aegilops caudata L. ×Triticum turgidum (L.) Thell

Interspecific hybrids and amphiploids between Aegilops caudata L. (2n = 2x = 14, CC) and Triticum turgidum (L.) Thell. conv. durum Desf M. K. (2n = 4x = 28, AABB) were produced. Such hybrids can be used to introduce desirable traits such as disease resistance into cultivated durum wheats. One of the durum parents was a ph I mutation of the cv. ‘Cappelli’ used for testing the possibility of direct introduction of alien variation into cultivated species. The amphiploids were obtained both through colchicine chromosome doubling and as natural non-reductional mciosis products. In both hybrids and amphiploids, meiotic pairing and fertility were studied. Hybrids showed varying degrees of pairing and, in addition to the one involving the ph 1 mutant, one high pairing hybrid was found (Ae. caudata× cv. ‘Capinera’). Cytological examination of microsporogenesis in amphiploids revealed a high frequency of bivalent formation. Fertility proved to be a very variable character since some of the amphiploids were almost completely sterile. The use of amphiploids in breeding programmes is discussed in relation to meiotic and fertility data.     

The influence of Elymus sibiricus L. genome on the diploidization system of wheat

Summary The meiotic behaviour of five tetraploid wheat strains x Elytricum fertile (2n=42 chromosomes, AABBD(SH) genomes) F₁ hybrids has been analysed. Multivalent associations were observed in the hybrids which could be attributed to Elymus sibiricus L. gene (s) somewhat suppressing the activity of the wheat homoelogous pairing control system. This interaction depends on the wheat genotype. The effect was particularly notable when Triticum turgidum var. salomonis was the wheat parent. The possibility of gene transfer from Elymus to wheat is discussed.     

Chromosome doubling and fertility study of Alstroemeria aurea × A. caryophyllaea

Self-pollinations of a diploid (2n = 2x = 16) interspecific hybrid Alstroemeria aurea × A. caryophyllaea, resulted in no seeds. Backcrosses of the hybrid with parent A. aurea did not produce any seeds. In an attempt to restore the hybrid fertility, an efficient in vitro procedure has been developed and applied effectively in the chromosome doubling of the diploid hybrid. Forty-one percent of the treated plants were proven to be truly tetraploid by chromosome counts and stomatal measurements after applying 0.2 to 0.6% colchicine for 6 to 24 hours. Over 87.5% of these colchicine-induced tetraploids were stable and retained their tetraploidy after one year of growth. The fertility of the hybrid was not restored although the pollen stainability was increased from 0 to 12% after chromosome doubling. Cytological studies on the pollen mother cells (PMCs) of the sterile diploid hybrids revealed abnormal meiotic behaviors. In addition, aneuploid chromosome numbers, ranging from 2n = 1 to 2n = 18, were observed in over 45% of the PMCs examined. PMCs of the colchicine-induced tetraploids showed that meiotic chromosome pairings were normal in most cases (1.59_I + 15.07_II + 0.05_III + 0.03IV). These results indicate that the sterility of this hybrid is not only caused by parental chromosome differences, but other complex fertility/sterility-regulating mechanisms are involved too. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genotypic control of chromosome pairing in amphiploids involving the cultivated oat Avena sativa L.

Summary A genotype of the diploid species Avena longiglumis (Cw 57) has been shown to modify the genetic control of diploid-like chromosome pairing in the cultivated oat, A. sativa (2n=6x=42) leading to increased homoeologous chromosome pairing in 4x hybrids between the two species (Rajhathy & Thomas, 1974). The Cw 57 genotype has a similar effect in increasing homoeologous chromosome pairing in amphiploids combining diploid and hexaploid genomes including associations between alien chromosomes and their corresponding pairs in hexaploid species. The effect of the Cw 57 genotype is probably in altering the specificity of chromosome pairing in the early stages of meiosis. The use of the Cw 57 genotype to induce homoeologous chromosome pairing as a technique for the transfer of desirable alien variation into the cultivated oat is discussed.     

Interspecific hybridisation of soybeans and perennial Glycine species indigenous to Australia via embryo culture

Summary Five sterile hybrids (2n=59) between a synthetic amphiploid of Glycine tomentella (2n=38) and G. caneseens (2n=40) as female and soybean cultivars Lincoln and Hark as males have been produced by embryo or ovule culture using transplanted endosperm. The hybrid plants are twining perennials like the female parent but possess a number of morphological characters which reflect the presence of the soybean genome. Indophenol oxidase isozymes from leaf extracts also provide good evidence of the hybrid nature of the cultured plants. These hybrids open the way for the exploitation of the diverse germplasm resources of the perennial Glycine species in soybean breeding.     

Wide hybridization between wheat (Triticum L.) and lymegrass (Leymus Hochst.)

A total of 240 F₁ hybrids was made beween wheat (Triticum aestivum L. em. Thell. (2n = 6x = 42) and T. carthlicum Nevski (2n = 4x = 28)) and perennial lymegrass (North European Leymus arenarius (L.) Hochst. (2n = 8x = 56) and North American L. mollis (Trin.) Pilger (2n = 4x = 28)). The wide crosses yielded embryos in 20% of caryopses and 96% of the embryos developed into normal hybrid plants. The hybrids were vegetatively vigorous, with evidence of the Leymus rhizomatous habit. Those deriving from L. arenarius survived overwintering in Iceland, but the hybrids L. mollis did not, whereas in a milder environment, both showed perenniality. Cytogenetic analysis of root tip cells before the plants were treated with colchicine showed that 21 out of 28 hybrids investigated had chromosome mosaics, with a population of both amphihaploid and amphidiploid cells. This spontaneous doubling of somatic chromosomes occurred in all cross combinations, with the highest average frequency of diploid cells (28%) in T. carthlicum × L. arenarius crosses. A few selfed seeds have been obtained from a T. aestivum × L. arenarius hybrid. All the hybrids were treated twice with colchicine, but the treatment appeared to have little or no effect on the frequency of chromosome doubling in the hybrids deriving from T. aestivum. The frequency of diploid cells, however, increased significantly (e.g. to 80%) in the hybrids deriving from the T. carthlicum parent. Genomic in situ hybridization confirmed the hybridity of the plants and showed that the hybrids were amphiploids containing genomes of both wheat and lymegrass. In situ hybridization using ribosomal DNA probe differentiated chromosomes of L. mollis, L. arenarius from those of wheat. The hybrids are being backcrossed with lymegrass pollen, aiming to domesticate the wild, perennial species. This revised version was published online in July 2006 with corrections to the Cover Date.     

A self-fertile trigeneric hybrid in the Triticeae involving Triticum,Hordeum, and Secale

Summary A self-fertile trigeneric hybrid in the Triticeae involving species from the Hordeum, Triticum and Secale genera has been produced. The trigeneric hybrid was obtained by crossing octoploid triticale (x Triticosecale Wittmack) with octoploid tritordeum (H. chilense × T. aestivum amphiploid). The trigeneric hybrid presented a genome constitution AABBDDRH^ch and 2n=8X=56 chromosomes. The cytogenetical analyses showed no chromosome instability nor homeologous pairing between Hordeum and Secale chromosomes. In the F₂ generation the chromosome number ranged from 42 to 52. Within this range, the plants with smaller numbers of chromosomes were more frequent. A preferential transmission of rye chromosomes could be inferred.     

Transfer of resistance to race 2 of Plasmodiophora brassicae from Brassica napus to cabbage (B. oleracea ssp. capitata). III. First backcross and F2 progenies from interspecific hybrids between B. napus and B. oleracea ssp. capitata

Summary The first backcross and F₂ progenies from triploid F₁ and tetraploid F₁ hybrids between B. napus and 2x and 4x B. oleracea ssp. capitata (cabbage) were studied for their general morphology, resistance to race 2 of the clubroot pathogen, chromosome number and meiotic chromosome behavior. No linkage was apparent between resistance and the major morphological characters. Unreduced gametes played a large part in the successful formation of seed of the B₁ and F₂ progeny. B₁ plants with low chromosome numbers were selected for use in recurrent backcrosses. The potential use of anther culture to extract gametic progenies from resistant B₁ and F₂ plants with higher chromosome numbers was suggested. The presence of homoeologous pairing observed in all the plants is considered advantageous for selecting suitable progeny in later generations.     

Cytological and genetical studies on hybrids between sorghum almum parodi (2n=40) and some diploid (2n=20) species of sorghum

Hybridization between S. almum (2n=40) and diploid (2n=20) species of Sorghum resulted in plants with 2n=40 and 2n=30 chromosomes. As segregation for many characters occurred in the offspring of those with 2n=40, hybrids of this type provide a means of transferring genes from the diploid to the tetraploid species of Eu-sorghum. Segregations for some genes in the progeny of these hybrids revealed heterozygosity in S. almum which may indicate that one of the ancestors of S. almum was a variety of S. vulgare very similar to the commercial grain sorghum.The triploids were only slightly fertile and the chromosome numbers of plants resulting from backcrossing to S. almum ranged from 30 to 46. Some of the plants with the higher chromosome numbers were self-fertile and segregation for genes which were present in the original diploid and tetraploid parents were observed in their off-spring. Backcrossing the triploid to the diploid parent produced fertile plants with 2n=20 and it is possible that the triploids could be used to transfer genes from the tetraploid to the diploid species of Eu-sorghum.The chromosome pairing in the triploids was similar to that expected in an autotriploid, but some non-homologous pairing was detected which may be the result of duplication of some chromosomes or chromosome segments within the genome (n=10) of S. vulgare.