Cytogenetic studies in Trifolium sp. related to berseem

Summary In a cytogenetic study of five annual Trifolium spp. the genetical relationships between (1) T. scutatum Boiss. and (2) T. plebeium Boiss., and between them and (3) T. carmeli Boiss., (4) T. echinatum M. B. and (5) T. latinum Seb., were studied in intra- and interspecific F₁ and F₂ hybrids. Germination, seedlings development, pollen fertility and chromosome configurations during meiosis and seedset were recorded. All the T. scutatum x T. plebeium F₁ were green, but the hybrids between either T. plebeium or T. scutatum and T. carmeli, T. echinatum or T. latinum were mainly albinos. Those that were green had a very high rate of mortality and the survivors produced many albino seedlings. Pollen fertility was about 60% in scutatum x plebeium F₁s, and one of their eight bivalents was heteromorphic. Hybrids between either of these and T. carmeli, T. echinatum or T. latinum had 30–50% stainable pollen and had several multivalents, suggesting the existence of a system of chromosomal translocations in these species. Seed set was roughly correlated with pollen stainability. The nature of the isolation mechanisms operating between these species is discussed.Contribution from the Volcani Center, Agricultural Research Organization, Bet Dagan, Israel. 1973 Series, No. 186-E. The work reported here was financed by Grant FG-IS-222 from the U.S. Department of Agriculture under P. L. 480.     

Cytogenetic studies in Trifolium Spp. related to berseem

Summary Seed formation by large-scale hybridization within and between Trifolium species related to T. alexandrinum L., was studied. The twelve species studied were: a. T. alexandrinum L., b. T. berytheum Boiss., c. T.salmoneum Mout., d, T. apertum Bobr., e. T. Meironense Zoh. et Lern., f. T. echinatum M.B., g. T. latinum Seb., h. T.carmeli Boiss., i. T. scutatum Boiss., j. T. plebeium Boiss., k. T. vavilovi Eig. and 1. T. constantinopolitanum Ser. Hybridization was done either by emasculation and fertilization by hand, or in nature, by utilizing the existence of natural stands and to serve as pollen source the self-incompatibility of some of the species concerned. Results of the two methods were highly comparable although seedset was much higher when crossing was done manually.Crossability, as estimated by seedset, varied in specific cross combinations and ranged between 0 and 70%. Based on the pattern of crossability, five crossability groups were identified with > 20% seedset in interspecific—intra-group cross combinations, and usually less than 5% in inter-group cross combinations. Species a, b, c, d and e form the first crossability group; f, g and h form the second one; h, i, j belong to the third crossability group, while k and l appear as unispecific fourth and fifth crossability groups. The high level of seedset in interspecific hybridization is discussed.Publication of the Agricultural Research Organization No. 2195-E. The work reported here was financed by Grant FG-IS-222 from the U.S. Department of Agriculture under P.L. 480.     

Karyotypes and serum transferrin patterns of hybrids between Asian and Oceanian black rats,Rattus rattus

Karyotypes and serum transferrin patterns were examined in Asian and Oceanian black rats (R. rattus). Japanese R. r. tanezumi and Malayan R. r. diardii had 2n=42, but Australian and New Guinea R. r. rattus showed 2n=38 chromosomes. F₁ hybrids between Japanese and Australian rats and Malayan and New Guinea rats had 2n=40 chromosomes which consists of the two genomes of both parents. Although various matings between the F₁ hybrids were made, only one F₂ male rat with 2n=39 chromosomes was obtained. The F₁ hybrids seem to be semisterile. Parental transferrin phenotypes were TfR in Japanese rats and TfCD in Oceanian rats. F₁ hybrids examined showed TfRD in both male and female and one F₂ hybrid had TfR type transferrin. Based on the above investigations, it is suggested that Asian and Oceanian black rats are geographically isolated and evolved different chromosomal and serum transferrin characteristics, but the sexual isolation of the two groups is incomplete at the present time.Contribution No. 826 from the National Institute of Genetics, Japan. Supported by a grant-in-aid from the Ministry of Education of Japan (Scientific Expedition in 1968, No. 8801 in 1969 and No. 9001 in 1970).     

Durum wheat as a candidate for the unknown female progenitor of bread wheat: an empirical study with a highly fertile F<Subscript>1</Subscript> hybrid with <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss.

Hexaploid bread wheat was derived from a hybrid cross between a cultivated form of tetraploid Triticum wheat (female progenitor) and a wild diploid species, Aegilops tauschii Coss. (male progenitor). This cross produced a fertile triploid F₁ hybrid that set hexaploid seeds. The identity of the female progenitor is unknown, but various cultivated tetraploid Triticum wheats exist today. Genetic and archaeological evidence suggests that durum wheat (T. turgidum ssp. durum) may be the female progenitor. In previous studies, however, F₁ hybrids of durum wheat crossed with Ae. tauschii consistently had low levels of fertility. To establish an empirical basis for the theory of durum wheat being the female progenitor of bread wheat, we crossed a durum wheat cultivar that carries a gene for meiotic restitution with a line of Ae. tauschii. F₁ hybrids were produced without using embryo rescue techniques. These triploid F₁ hybrids were highly fertile and spontaneously set hexaploid F₂ seeds at the average selfed seedset rate of 51.5%. To the best of our knowledge, this is the first example of the production of highly fertile F₁ hybrids between durum wheat and Ae. tauschii. The F₁ and F₂ hybrids are both similar morphologically to bread wheat and have vigorous growth habits. Cytological analyses of F₁ male gametogenesis showed that meiotic restitution is responsible for the high fertility of the triploid F₁ hybrids. The implications of these findings for the origin of bread wheat are discussed.     

Chromosomal location of a pollen fertility-restoring gene, Rf, for CMS in Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of A. galanthum Kar. et Kir. revealed by genomic in situ hybridization

In a previous study, we developed cytoplasmic male sterile lines of Allium fistulosum possessing the cytoplasm of A. galanthum, a wild species, by continuous backcrossing. Furthermore, we reported the presence of a pollen fertility-restoring gene (Rf) for cytoplasmic male sterility (CMS) in A. fistulosum from segregation of pollen fertility of backcross progenies. In the present study, genomic in situ hybridization (GISH), using genomic DNA of A. galanthum as the probe DNA and that of A. fistulosum as the blocking DNA, was applied to F₁ hybrids between both species and backcross progenies to determine the chromosomal location of the Rf locus. By means of GISH, eight chromosomes from A. galanthum were clearly discriminated from those of A. fistulosum in the F₁ hybrids, and chromosome substitution process through continuous backcrossing was visualized. Interestingly, the chromosome region from A. galanthum, specific to male fertile plants, was detected in one chromosome of BC₄ to BC₇ generations. Based on the karyotype analysis of the male fertile plants, the chromosome was identified as the 5F chromosome. Our results confirm that the Rf locus is located on the 5F chromosome of the male fertile plants. This is the first report that identified the chromosomal location of the pollen fertility-restoring gene in A. fistulosum.     

Reproductive interactions between hybridizing irises: analyses of pollen-tube growth and fertilization success

Pollen-tube growth and seed siring ability were measured in crosses between the Louisiana iris species Iris fulva and Iris hexagona and their F₁ and F₂ hybrids. Flowers of the parental species were pollinated with self, outcross intraspecific, and interspecific pollen. Pollen-tube lengths were similar for all three pollen types in I. fulva, but in I. hexagona interspecific pollen tubes were longer than intraspecific pollen tubes. Pollen-tube lengths also differed for F₁ and F₂ flowers pollinated with I. fulva, I. hexagona, and hybrid pollen. For both hybrid classes I. fulva pollen tubes were the shortest while pollen tubes from I. hexagona and hybrids grew the furthest. Mixtures of genetically marked pollen were used to assess the seed siring ability of intra- and interspecific pollen in the parental species by varying the proportion of each pollen type in a replacement series design. For both species, the observed proportions of hybrid seeds were lower than the expected based on the frequency of each pollen type in the mixtures across all treatments. Flowers of I. fulva produced less than 10% hybrid progeny even when 75% of the pollen applied to stigmas was derived from interspecific flowers. The frequency of hybrid seed formation was somewhat greater in I. hexagona, but was still significantly lower than expected across all mixture treatments. Seed set per fruit remained constant across the mixture treatments for both species, but in I. fulva fruit set decreased with an increase in the proportion of interspecific pollen. The data indicate that both pre- and postfertilization processes contribute to discrimination against hybrid seed formation.     

Development of a fertile genetic bridge between Trifolium ambiguum M. Bieb. and T. repens L.

  Trifolium ambiguum M. Bieb and T. repens L. are taxonomically related but very difficult to cross. The rare hybrids so far reported between these two species were obtained only by embryo culture. This difficulty has been overcome in the present research by the creation of a “fertile bridge” between T. ambiguum and T. repens. Characters of interest can now be transferred from T. ambiguum to T. repens by using this “fertile bridge” without the use of sophisticated techniques. An array of backcross progenies was generated from crosses between a T. ambiguum×T. repens F₁ hybrid (8x H-435) and its parental species. The 8x hybrid was cross-fertile only with T. repens and resulted in 145 seeds from 1578 reciprocal crosses. Eleven of nineteen initially grown BC₁F₁ plants were all hexaploid with an average pollen stainability of 41.6%. A high frequency of multivalents at metaphase-I indicated that both autosyndetic and allosyndetic pairing occurred. Backcrosses of 6x BC₁F₁ plants to T. repens resulted in 5x BC₂F₁ plants with an average pollen stainability of 59.3%. On the other hand, 6x BC₁F₁×6x T. ambiguum crosses did not produce any seed and only two pentaploid plants were obtained from 6x BC₁F₁×4x T. ambiguum crosses. The difficulty encountered in generating 6x backcross progeny with 6x T. ambiguum was overcome by intercrossing the 6x BC₁F₁ plants and producing 6x BC₁F₂ plants with an average pollen stainability of 65.8%. One of these 6x BC₁F₂ plants was cross-compatible as a female with 6x T. ambiguum and resulted in CBC₂ plants that were all cross-compatible with 6x T. ambiguum. The 6x BC₁F₂ plants are likely to be superior to 6x BC₁F₁ progeny, as they have exhibited better expression of the combined rhizomatous and stoloniferous growth habit, improved fertility, more frequent nodal rooting and heavier nodulation. Consequently, the 6x BC₁F₂ plants can either be used directly in the selection programme or as a “fertile bridge” between the two parental species. The present work has resulted in the development of a series of fertile hybrids by the manipulation of chromosome numbers, combining the agronomic characteristics of the parent species in varying genome balances and at a range of ploidy levels. It is concluded that the initial sterility of the primary interspecific hybrids need not be a barrier to successful inter-breeding. Received: 2 August 1996 / Accepted: 4 April 1997     

Wide hybridization in Brassica

Summary Crosses were made to obtain interspecific hybrids between B. fruticulosa (wild species , 2n = 16) × B. campestris (cultivar , 2n = 20). Although many pollen grains germinated and their tubes entered the style, only about 30% of the ovules received pollen tubes. Fertilized ovules aborted at various stages of development. A few hybrid seeds resulted from hand pollinations in the field, and they showed poor germination and seedling establishment. The in vitro culture of ovaries, ovules, and seeds increased the frequency of obtaining hybrid seeds and plants: the most effective method was ovary culture followed by ovule culture. The hybrid nature of the plants was confirmed through morphological, cytological, and electrophoretic studies. A meiotic analysis of F₁ hybrids (2n = 18) showed that they had 0–5 bivalents and were completely pollen sterile. Electrophoretic analysis of leaf esterases and acid phosphatases of F₁ hybrids revealed bands derived from each parent. Induced amphidiploids of F₁ hybrids contained mostly bivalents, and had about 50% fertile pollen.     

Occurrence of genetic tumours in Triticum interspecies hybrids

Summary F₁ interspecific hybrids involving nine tetraploid Triticum species were studied. Some developed leaf tumours at the seedling stage. Tumorous hybrids were restricted to crosses involving either T. timopheevi or T. araraticum as one parent. The hybrids from the rest of the crosses, including those of T. timopheevi × T. araraticum, were non-tumorous. Genetically divergent and non-integrated parental species appeared to be inducing spontaneous tumour formation in their hybrids.     

Airborne‐pollen pool and mating pattern in a hybrid zone between Pinus pumila and P. parviflora var. pentaphylla

The reproductive isolation barriers and the mating patterns among Pinus pumila, P. parviflora var. pentaphylla and their hybrids were examined by flowering phenology and genetic assays of three life stages: airborne‐pollen grains, adults and seeds, in a hybrid zone on Mount Apoi, Hokkaido, Japan. Chloroplast DNA composition of the airborne‐pollen was determined by single‐pollen polymerase chain reaction. Mating patterns were analysed by estimating the molecular hybrid index of the seed parent, their seed embryos and pollen parents. The observation of flowering phenology showed that the flowering of P. pumila precedes that of P. parviflora var. pentaphylla by about 6 to 10 days within the same altitudinal ranges. Although this prezygotic isolation barrier is effective, the genetic assay of airborne‐pollen showed that the two pine species, particularly P. pumila, still have chances to form F₁ hybrid seeds. Both parental species showed a strong assortative mating pattern; F₁ seeds were found in only 1.4% of seeds from P. pumila mother trees and not at all in P. parviflora var. pentaphylla. The assortative mating was concluded as the combined result of flowering time differentiation and cross‐incompatibility. In contrast to the parental species, hybrids were fertilized evenly by the two parental species and themselves. The breakdown of prezygotic barriers (intermediate flowering phenology) and cross‐incompatibility may account for the unselective mating. It is suggested that introgression is ongoing on Mount Apoi through backcrossing between hybrids and parental species, despite strong isolation barriers between the parental species.     

Natural hybridization in JapaneseCalamagrostis

Morphological and pollen studies were made for the collections belonging toCalamagrostis langsdorffii, C. Sachalinensis, C. longiseta, C. nana and various intermediates between these species from central Honshu, the chromosome numbers of which were reported in a previous paper (Tateoka, 1976). Additional collections made from the Akaishi Range in 1976 were also subjected to the present work. The intermediates showed 0%, or nearly 0%, pollen stainability and a mosaic of morphological features of putative parental taxa which can be estimated to occur for F₁ hybrids. This estimation was sustained by a comparison with the pattern of character expressions for the artificially raised F₁ hybrids of various European species ofCalamagrostis as reported by Nygren (1962). These results as well as field observations strongly suggested that the intermediates may be wholly or almost wholly F₁ hybrids. The hybrids were sometimes found in abundance within a limited area, but seemed to have little capacity to migrate from the places where they originated. The following combinations, which were not previously recorded, were disclosed:C. longiseta×C. sachalinensis, C. langsdorffii×C. nana, C. langsdorffii×C. longiseta, C. longiseta×C. nana. Possibilities of introgression in these hybridizing species were discussed. The hybrids were tetraploids except one hexaploid which was discovered in a mixed population ofC. nana, C. longiseta and their putative F₁ hybrids at the tetraploid level. The hexaploid in question, which seemed to have resulted from the participation of an unreduced gamete, was very similar morphologically toC. nana subsp.hayachinensis distributed in the mountain far distant from central Honshu. Geographical distributions of the hybrid plants were surveyed through examination of the herbarium specimens.     

Crossability of Brassica tournefortii and B. rapa, and morphology and cytology of their F1 hybrids

The crossability between Brassica tournefortii (TT, 2n = 20) and Brassica rapa (AA, 2n = 20) and the cytomorphology of their F₁ hybrids were studied. Hybrids between these two species were obtained only when B. tournefortii was involved as a female parent. The hybrid plants were intermediate for most of the morphological attributes and were found to be free from white rust under field conditions. The F₁ plants showed poor pollen fertility, although occasional seed set was achieved from open pollination. Self-pollination or backcrosses did not yield any seeds in these plants. The occurrence of chromosome association ranging from bivalents (0–7), trivalents (0–2) to a rare quadrivalent (0–1) in the dihaploid hybrids indicates pairing between the T and A genomes. The homoeologous pairing coupled with seed set in the F₁ plants offer an opportunity for interspecific gene transfers from B. tournefortii to B. rapa and vice-versa through interspecific hybridization. Received: 3 July 2000 / Accepted: 22 September 2000     

Chromosomal rearrangements directly cause underdominant F1 pollen sterility in Mimulus lewisii–Mimulus cardinalis hybrids

Chromosomal rearrangements can contribute to the evolution of postzygotic reproductive isolation directly, by disrupting meiosis in F₁ hybrids, or indirectly, by suppressing recombination among genic incompatibilities. Because direct effects of rearrangements on fertility imply fitness costs during their spread, understanding the mechanism of F₁ hybrid sterility is integral to reconstructing the role(s) of rearrangements in speciation. In hybrids between monkeyflowers Mimulus cardinalis and Mimulus lewisii, rearrangements contain all quantitative trait loci (QTLs) for both premating barriers and pollen sterility, suggesting that they may have facilitated speciation in this model system. We used artificial chromosome doubling and comparative mapping to test whether heterozygous rearrangements directly cause underdominant male sterility in M. lewisii–M. cardinalis hybrids. Consistent with a direct chromosomal basis for hybrid sterility, synthetic tetraploid F₁s showed highly restored fertility (83.4% pollen fertility) relative to diploids F₁s (36.0%). Additional mapping with Mimulus parishii–M. cardinalis and M. parishii–M. lewisii hybrids demonstrated that underdominant male sterility is caused by one M. lewisii specific and one M. cardinalis specific reciprocal translocation, but that inversions had no direct effects on fertility. We discuss the importance of translocations as causes of reproductive isolation, and consider models for how underdominant rearrangements spread and fix despite intrinsic fitness costs.     

NATURAL AND ARTIFICIAL HYBRIDS OF HELIANTHUS MAXIMILIANI X H. GROSSESERRATUS

Long , Robert W. (Ohio Wesleyan U., Delaware.) Natural and artificial hybrids of Helianthus Maximiliani × H. grosseserratus. Amer. Jour. Bot. 46(10): 687–692. Illus. 1959.—An investigation of the occurrence of natural hybridization in two perennial sunflowers, Helianthus Maximiliani and H. grosseserratus, was begun in 1950. Subsequently, artificial F₁, F₂, and first and second backcross generations were produced. Fertility and vigor were high in all these plants, but F₁ plants appeared to excel the others in these characteristics. Observations in the experimental garden were supplemented by examination of chromosomes in pollen mother cells, comparisons of herbarium collections, and study of wild populations. Evidence pointed to close genetic relationship of the species and to the occurrence of natural hybridization in areas of distributional overlap. In 1957 and 1958, field work in these areas resulted in the scoring of 18 natural populations, 3 of which consisted of both parental species plus putative F₁ hybrids. Two explanations are offered to account for the seeming absence of introgression. The results support the conclusion that natural hybridization leads to the establishment of F₁ hybrids and that introgression does not occur to any significant extent. Although both species display a high degree of interfertility, they are distinct morphologically. For this reason, it is advisable to maintain them as separate species.     

RELATIONSHIP OF WOODY PARTHENIUM ARGENTATUM AND HERBACEOUS P. HISPIDUM VAR. AURICULATUM (ASTERACEAE)

Interspecific hybrids between woody Parthenium argentatum Gray (guayule), native to Mexico and Southwest Texas, and herbaceous perennial P. hispidum var. auriculatum (Britton) Rollins, native to the United States, were obtained successfully. The F₁ hybrids were intermediate for most morphological characters with the exception of the short woody stem, yellow pollen color, and the trichome morphology. Chromosome counts revealed the presence of 2n = 36 A-chromosomes in P. argentatum. The same number of A-chromosomes and four B-chromosomes were found in P. hispidum var. auriculatum. Observations of pollen mother cells showed regular meiosis in both parental species. At diakinesis, chiasmata averaged 1.12 and 1.24 per bivalent for P. argentatum and P. hispidum var. auriculatum, respectively. Meiotic behavior of the F₁ hybrids was irregular. F₁ hybrids averaged 4.43 univalents at metaphase I, 1.95 laggards at anaphase I, and 1.62 micronuclei at the tetrad stage. The low pollen stainability (5.1%) in the F₁ hybrids and the limited number of viable BC₁ seeds (4.07%) may be reflections of the irregular meiosis. Although these primary hybrids are partially fertile, they can be used to introduce desirable characteristics of P. hispidum var. auriculatum, such as herbaceous perennial habit, regrowth ability, and cold tolerance into guayule.     

The association between pollen size and Renner complex in Oenothera villaricae and O. picensis ssp. picensis and their hybrids: evidence for preanthesis pollen competition

In both Oenothera villaricae Dietrich and O. picensis ssp. picensis, chromosomes are transmitted as two Renner complexes. Reciprocal combinations of the Renner complexes produce eight different F₁ hybrids, but only seven are viable. Each species, and each F₁ hybrid, produces three sizes of pollen, approximately 50% small sterile grains, 15% medium-sized viable grains and 35% large viable grains. Medium- and large-sized grains were separated manually and subjected to random amplified polymorphic DNAs (RAPDs) analysis. A pattern of RAPD amplifications was obtained which indicates that, for each species and F₁ hybrid, one specific Renner complex characterizes the medium- and another the large-sized viable pollen. The results indicate that pollen size is determined in part by the pollen genotype and in part by the genotype of the other pollen grains developing within the same anther.     

A study of chromosome association in hybrids between diploid species of Festuca

Hybrids between the two diploid species Festuca donax and F. drymeja had regular chromosome association, forming 7 bivalents at metaphase I. However, when the two species were crossed with a third species F. scariosa, the hybrids involving F. drymeja showed greater desynapsis than those with F. donax. When the F₁ hybrid F. donax × F. drymeja was crossed with F. scariosa, the trispecific progeny could be grouped into three classes according to the degree of desynapsis recorded. Abnormalities associated with the fusion of pollen mother cells, which produced highly polyploid cells, were also observed in the trispecific hybrids. Failure of chromosomal pairing and the occurrence of syncytes is attributed to genotypic interactions.     

THE RELATIONSHIPS BETWEEN MALE AND FEMALE FERTILITY AND AMONG TAXA IN DIPLOID WHEATS

Pollen stainability appears to be a reliable indication of the ultimate seed set in diploid interspecific hybrid and backcross populations in Triticum L. The correlation between percent pollen stained and number of seeds set is positive and highly significant (r = 0.92). Estimates of male and female fertility in the hybrids and backcrosses are interpreted to indicate that the domesticated diploid Triticum monococcum L. and wild diploid T. boeoticum Boiss. em. Schiem, are one and the same species, and that T. urartu Tum. is not a variety of monococcum or boeoticum, but rather a separate species. The F₁ hybrids and backcrosses between monococcum and boeoticum are normally male and female fertile. The F₁ hybrids between monococcum and urartu are completely sterile and complete to partial sterility exists in backcrosses.     

Genetic interactions between wheat and rye genomes in triticale

Summary Six primary triticale lines were produced from two advanced breeding lines of Triticum durum and three inbred genotypes of Secale cereale. The wheat and rye parents and the triticale derivatives were crossed in all possible combinations within each species group. Chiasma and univalent frequency of parents and hybrids were determined. The primary triticale lines had more univalents and less chiasmata per pollen mother cell than the corresponding wheat and rye parents together. The parental wheat F₁ exhibited negative heterosis for chiasma frequency whereas all rye hybrids had much higher chiasma frequencies than their inbred parents. Triticale F₁s generally showed lower chiasma frequencies and more univalents than their parents, but the degree of pairing failure was dependent upon which of the parental species within the triticale, wheat or rye, was in the heterozygous state. F₁s with heterozygous wheat genome only showed the least reduction in chiasma number (presumably caused by gene actions within the wheat genome), while F₁s with heterozygous rye genome showed high reduction in chiasma frequency and an increase in pairing failure (induced by negative interactions between the heterozygous rye and the wheat genome in triticale). A high correlation was found between the frequency of undisturbed pollen mother cells and the frequency of aneuploids in the subsequent generation. A higher number of aneuploids occurred in those populations which were heterozygous for the rye genome.