Interspecific reproductive barriers and genomic similarity among the rough-seeded Lupinus species

The nature of reproductive barriers and meiotic behaviour of chromosomes were studied in interspecific hybrids among the six rough-seeded Lupinus species. Out of 30 different interspecific crosses attempted in all possible combinations, eight produced viable F, seeds. These successful crosses involved L. cosentinii, L. digitatus, L. atlanticus and L. pilosus. Crosses of L. princei with other species resulted in shrivelled F, seed in all combinations. In contrast to previous reports, crosses of L. palaestinus with all other rough-seeded lupins were incompatible as no F₁ hybrids were obtained. Barriers to interspecific reproduction were identified as nucleo-cytoplasmic and embryo-endosperm incompatibility in unsuccessful crosses, and chromosomal imbalances in F₁ hybrids. Gene transfer is possible among L. cosentinii, L. digitatus and L. atlanticus, which produced F₂ seed. Patterns of chromosome configurations in F₁ meiotic cells suggested that these species have at least two genomes partially in common, but include inversions and translocations. The genome of L. atlanticus is closer to L. digitatus than to L. cosentinii, and that of L. pilosus is closer to L. atlanticus than to L. cosentinii. L. princei appears to have an isolated genome within the rough-seeded Lupinus species.     

The production and behaviour of hexaploid hybrids between Hordeum vulgare and H. Bulbosum

Summary To produce hexaploid (or other polyploid) hybrids, diploid or tetraploid Hordeum vulgare was crossed with hexaploid or octoploid H. bulbosum, and perennial triploid hybrids between the two species were treated with colchicine. The crosses did not yield viable plants: seedset was low, the seed aborted and embryo culture was unsuccessful. The colchicine treatments geve rise to plants in which hexaploid chromosome numbers were observed. At the hexaploid level chromosomal instability occurred, resulting in chromosome elimination.The colchicine-treated triploid hybrids showed in the first years after the treatment better fertility after open flowering than untreated plants, but the level of fertility remained very low. The offspring consisted of haploid, diploid and approximately triploid plants like H. vulgare, tetraploid and approximately tetraploid plants like H. bulbosum, and plants with hybrid morphology and unstable chromosome number, which were highly sterile. Thus the crossing barrier between H. vulgare and H. bulbosum could not be broken down at higher ploidy level.     

Ploidy Manipulation and Introgression of Resistance to Alternaria Helianthi from Wild Hexaploid Helianthus Species to Cultivated Sunflower (H. annuus L.) Aided by Anther Culture

The present investigation discusses the scope for transferring of resistance to leaf spot disease incited by Alternaria helianthi from two hexaploid wild species (H. tuberosus and H. resinosus) to diploid cultivated sunflower. Interspecific hybrids produced between sunflower and these two hexaploid species were partially fertile with tetraploid chromosome status. Backcrosses of these interspecific hybrids with cultivated sunflower resulted in the formation of sterile triploid plants. To overcome the problem of sterility and facilitate backcrosses with cultivated sunflower, anther culture of the tetraploid interspecific hybrids was carried out to bring down their chromosome number to diploid status. Anthers from both interspecific hybrids were cultured on basal Murashige and Skoog media supplemented with varying concentrations of organics and the growth regulators benzyladenine and naphthaleneacetic acid. Anthers of interspecific hybrids involving H. resinosus responded well and regenerated through an embryogenic route at a frequency of 98.7%. But in interspecific hybrids with H. tuberosus, anthers formed callus and subsequently regenerated shoots through an organogenic pathway. DNA ploidy analysis of anther culture plants of interspecific hybrids derived from H. tuberosus crosses was carried out to identify plants with desired diploid status. In vitro screening of parents, interspecific hybrids and anther culture plantlets against A. helianthi showed resistance in 68.5% of the anther culture plants of interspecific hybrids from H. tuberosus and in 24.3% of the plants derived from interspecific hybrids involving H. resinosus.     

Pollen grain germination and pollen tube growth following in vivo and in vitro self and interspecific pollinations in annual Cicer species

Summary Germination of pollen grains and growth of pollen tubes were studied to determine the cause of barreness in crosses among annual Cicer species. In vivo and in vitro time-course studies and fluorescent microscopy revealed no pollination incompatibility among the selfs, crosses and reciprocals of C. arietinum L., C. reticulatum Lad. and C. cuneatum Rich. In general, Cicer pollen grains germinated and grew on styles of Cicer species. Pollen tube growth was characterized by irregularly spaced and intermittent callose deposits. Failure of seed formation in interspecific pollinations may be attributed to the slowness of pollen tube growth or collapse of fertilized ovules. In addition to these causes, shortness of stamens and sparsity of pollen grains were responsible for flower drop in natural selfs. Although the number of pollen tubes entering the micropyle in interspecific pollinations was low, it may be possible to grow the fertilized ovules on an artificial medium to obtain F₁ plants.     

Interspecific hybrid of Helianthus annuus × H. simulans: Characterization and utilization in improvement of cultivated sunflower (H. annuus L.)

The first success at interspecific hybridization between cultivated sunflower(H. annuus) and a diploid perennial species, H. simulans is reported. The F₁s from both direct and reciprocal directions exhibited dominance of the wild species phenotype and were pollen sterile. Meiosis was irregular in the F₁ hybrids and both univalents and multivalents were observed. Multiplication of the interspecific hybrids was achieved through in vitro culture of nodal sections on Murashige & Skoog medium supplemented with 0.5 mg l^-1 benzyladenine. Fertility of the interspecific hybrids was improved by subjecting the in vitro proliferating shoots to 0.001% colchicine incorporated in the shoot multiplication medium. The amphiploids serve as fertile bridges and facilitate interspecific gene transfer through conventional breeding. This revised version was published online in July 2006 with corrections to the Cover Date.     

Meiotic behavior in interspecific hybrids between Brachiaria ruziziensis and Brachiaria brizantha (Poaceae)

The meiotic behavior of two half-sib interspecific tetraploid (2n = 4x = 36) promising hybrids, a sexual and an apomictic one, from crosses B. ruziziensis and B. brizantha, was evaluated. Although chromosome paired predominantly as bivalents, a few tri- and quadrivalents were recorded. Results suggest that B. brizantha and B. ruziziensis are closely related and genetic recombination is expected in hybrids. Introgression of specific target genes from B. ruziziensis into B. brizantha and vice-versa may be foreseen. However, abnormalities such as irregular chromosome segregation, chromosome stickiness and abnormal cytokinesis reported in these hybrids affect pollen fertility. More than 65% of pollen grains are sterile. Since the distinctive cytological feature of these hybrids is abnormal cytokinesis, this fact suggests that both parental genomes are unable to coordinate their activities with regard to this cytological phenomenon. Deployment of such hybrids in the process of developing varieties is discussed.     

How wide can a wide cross be?

Summary Wide crosses in wheat are reviewed in relation to various factors that facilitate wide crossing to show that wide crosses can be as wide as one can make them. Included in this review is a particular reference to wheat-wheatgrass (Agropyron complex) crosses and an update on wide crosses of wheat with various genera of Agropyron complex. Hybrid seed set is too variable to predict whether a wide hybrid, where no seed was obtained in one attempt, will not be possible. High crossability genes seem to facilitate not only fertilization but also seed development, enabling embryo rescue. Variability for crossability occurs not only in wheat but also in alien species. Contrary to conventional thinking, several wide hybrids with wheat can be produced when species with lower chromosome numbers are used as female parents. Pre-and post-fertilization barriers to wide crosses do not appear to be equally strong, and can be overcome by the development and application of various technologies. Considering these aspects of wide hybridization, and based on recent successes in producing previously unsuccessful and very wide hybrids, it is concluded that how wide cross between plant species can be made is an open question and that many new and wider hybrids can be produced in future.     

Incongruity of interspecific and intergeneric crosses involving Nicotiana and Petunia species that exhibit potential for somatic hybridization

Summary Extensive pollinations were made in attempts to produce selected intergeneric or interspecific hybrids using Nicotiana and Petunia species that exhibit potential for somatic hybridization. Pollination between pairs of species was done by standard and bud-pollination methods. Nicotiana alata or N. tabacum when pollinated reciprocally with selected Petunia species failed to produce hybrids. Likewise, no interspecific hybrids were obtained between Petunia parviflora and 4 Petunia species or P. hybrida. The type of incongruity existing between these species is discussed in relation to the production of hybrids by standard In vitro techniques as compared to somatic hybridization.On leave from the Department of Horticulture, Michigan State University, E. Lansing, Michigan, USA. supported by a European Molecular Biology Organization (EMBO) fellowship and The Fred C. Gloeckner Foundation, N. Y., USA.Supported by the Agricultural Research Council.     

Polyploidy and sexual polyploidization in the genus Vaccinium

Genus Vaccinium,consisting of blueberries, cranberries, lingonberries and many related wild species, includes diploid, tetraploid, and hexaploid species. Most evidence indicates that the tetraploid species are autotetraploids, with non-preferential bivalent chromosome pairing. Although homoploid interspecific crosses usually produce numerous fertile hybrids if the parents are from the same section of the genus, inter sectional crosses at the diploid level normally produce no seedlings, weak seedlings, or seedlings that are very low in fertility. There is a strong but not complete triploid block within Vaccinium. Even with insections, tetraploid × diploid (and the reciprocal) crosses normally give only a few tetraploid and a few triploid hybrids. Hexaploid × diploid crosses within sections are very hard to make, and the few hybrids that have been obtained are pentaploid. The frequency of 2n gametes varies,both among genotypes within species and among species. Vaccinium pollen is normally shed in tetrads, and the frequency of large pollen grains shed in dyads can be used to estimate 2n gamete frequency. Cultivated blueberries occur at both the tetraploid and the hexaploid levels, and there are important genetic resources in the diploids. Unreduced gamete production has permitted transfer of genetic material from the diploid to the tetraploid level and from the diploids and tetraploids to the hexaploid level via triploid hybrids. Intersectional crosses can occasionally produce tetraploid Vaccinium hybrids that appear to behave as amphidiploids and have medium to high fertility. CommercialVaccinium cultivars are normally propagated by cuttings. Intersectional hybridization, chromosome doubling, and asexual propagation could permit the production of novel hybrid combinations with value as ornamentals or in fruit production. This revised version was published online in July 2006 with corrections to the Cover Date.     

Improvement in hydroxy fatty acid seed oil content and other traits from interspecific hybrids of three Lesquerella species: Lesquerella fendleri , L. pallida, and L. lindheimeri

Interspecific hybridization offers potential to improve the hydroxy fatty acid (HFA) content of lesquerella. Lesquerella fendleri is currently being developed for cultivation as a potential new industrial oilseed crop because of its seed productivity. However, it has lower HFA content compared to L. pallida and L. lindheimeri. The objectives of this research were to improve the HFA oil content of L. fenderi through interspecific hybridization and to characterize hybrids and successive generations for seed oil fatty acid profile, fertility, seed set and other morphological traits. In this work, three species were successfully hybridized, self-pollinated, and backcrossed. Ovule culture was used in some cases to produce progeny where interspecific hybrids did not produce viable embryos. The traits measured were petal length, ovules per silique, seeds per silique, and weight of 1000 seed. Patterns of leaf trichomes were used to distinguish between parents and hybrids. Seed per silique indicated that autofertility occurred in L. pallida but not in the other two species. HFA oil content of L. fendleri seed were 50.5% compared to 80 and 84% for L. pallida and L. lindheimeri, respectively. The HFA oil content of the hybrids ranged from 57 to 70% in A₂ and A₃ generations, and the range of values depended on the parents used in the crosses. These measurements will help predict the value of different interspecific crosses for breeding. Segregation for various yield related traits should allow selection for favorable improvements in the HFA trait and in seed yield.     

Fertilisation assessment and postzygotic development in several intra- and interspecific Prunus hybrids

Summary the clearing technique was applied to assess fertilisation and to follow the postzygotic development in ten intra- and interspecific crosses among Prunus species used as rootstocks. The technique allowed to distinguish fertilised ovules from unfertilised ones, as well as to calculate the fertilisation percentage in intra-and interspecific compatible crosses. All the stages from unfertilised to fertilised ovules were analysed in order to determine the best time for embryo rescue. Differences in postzygotic development among the crosses are discussed in terms of the genotype of the seed parent in the pollen-pistil interactions.Abbreviations DAP Days After Pollination     

The origin of lentil and its wild genepool

Summary Following hybridization experiments and cytogenetic analysis of interspecific hybrids three chromosome interchanges were found between the cultivated lentil L. culinaris and L. nigricans, and only one between the cultivated species and L. orientalis. This indicates that the latter species is more likely to be wild progenitor of lentil. The partial fertility of the interspecific hybrids indicate further that both L. nigricans and L. orientalis should be included in the wild genepool of lentil, and their variation can be exploited by relatively simple hybridization techniques. The wild lentils L. orientalis and L. nigricans are morphologically very similar but reproductively strongly isolated from one another by the albino seedling of their hybrids. It has been suggested that the populations of L. orientalis that gave rise to the cultivated lentil still possess a similar chromosome arrangement as in L. culinaris and are also capable of forming normal hybrids with L. nigricans. According to these considerations it is unlikely that lentil originated from populations at the south western corner of the distribution area of L. orientalis.     

Production of interspecific hybrids between <Emphasis Type="Italic">Lilium longiflorum</Emphasis> and <Emphasis Type="Italic">L. lophophorum</Emphasis> var. <Emphasis Type="Italic">linearifolium</Emphasis> via ovule culture at early stage

Interspecific hybridization was carried out between Lilium longiflorum and L. lophophorum var. linearifolium by using the cut style method of pollination, as a contrast, intraspecific hybridization between L. longiflorum ‘Gelria’ and L. longiflorum was also made, but no mature seeds and offspring were obtained from the two combinations under in vivo condition. Ovules excised from each carpel 5–35 days after pollination (DAP) were cultured on B5 or half-strength B5 medium containing sucrose at different concentrations in vitro. In L. longiflorum × L. lophophorum var. linearifolium, only 1.17% of ovules excised at 10 DAP developed into seedlings, and in L. longiflorum ‘Gelria’ × L. longiflorum, only 0.99% of ovules excised at 25 DAP developed into seedlings; none of the ovules excised at other different DAP in the two cross combinations produced any seedlings. The results showed that interspecific hybridization had a more serious post-fertilization barrier than the intraspecific hybridization, and that a lower concentration (3%) of sucrose led to better embryo development and higher percentage of seedlings in ovule cultures. All hybrid seedlings obtained were successfully transplanted to soil and grew normally. The progenies investigated were identified as true hybrids based on inter-simple sequence repeat (ISSR) analysis.     

Cytogenetics of interspecific hybrids in the genus Capsicum L.

Summary Crosses between a wild species C. chacoense and three cultivated species of chili pepper viz. C. annuum, C. frutescens and C. chinense yielded hybrids when C. chacoense was the seed parent but the reciprocal crosses were unsuccessful. C. chacoense × C. annuum F₁ hybrids were partly fertile and therefore an F₂ population could be raised; the other two F₁ hybrids were totally sterile. Chromosome pairing in the F₁ plants resulted largely in bivalents and a few multivalents and univalents. The genomes of the four species share large homologies and the role of chromosome structural changes in genome differentiation is suggested. Hybrid sterility is the major reproductive isolation mechanism.     

Use of the gene pools of Triticum turgidum ssp. dicoccoides and Aegilops squarrosa for the breeding of common wheat (T. aestivum), through chromosome-doubled hybrids

Summary Triticum turgidum ssp. dicoccoides (wild emmer wheat, AABB, 2n=28) and Aegilops squarrosa (goat grass, DD, 2n=14) comprise a rich reservoir of valuable genetic material, which could be useful for the breeding of common wheat (T. aestivum, AABBDD, 2n=42). Many accessions of both wild species, most of them selected for resistance to stripe rust, were used to make amphiploids. Two strategies were applied: (1) the production of autopolyploid cytotypes of the wild species, followed by hybridisation, and (2) the production of allotriploid interspecific hybrids, followed by doubling of the number of chromosomes. The first route was unsuccessful because of failure of the crosses between the autopolyploid cytotypes, possibly due to incongruity between the two species and to reduced fertility in the autopolyploid cytotypes. The second route yielded the desired synthetic hexaploids. However, the rate of success of the crosses was low and there were great differences between years, and within years between crosses. Embryo rescue was applied to obtain the primary hybrids (2n=21), which were highly sterile and had on average 0.3 bivalents and 20.4 univalents per pollen mother cell. Various abnormalities were recorded. Doubling of the number of chromosomes sometimes occurred spontaneously or was brought about by colchicine treatment. The large scale of the interspecific hybridisation programme ensured that one-third of the female and one-sixth of the male accessions were represented in the synthetic hexaploids.     

Pollen germination and pollen tube growth following self-pollination and intra- and interspecific pollination of Medicago species

Summary Pollen germination and pollen tube growth after self- and intra- and interspecific pollinations of annual [M. scutellata (L.) Mill., M. disciformis DC., and M. rigidula (L.) All.] and perennial (M. sativa L.) Medicago species were studied using a Carl Zeiss microscope with a fluorescence filter attachment.Self-pollination of self-fertile annual and intraspecific pollination of cross-fertile perennials were characterized by high pollen germination and rapid normal pollen tube growth. In contrast pollen germination percentages were very low and many pollen tube growth abnormalities occurred following interspecific pollination and self-pollination of self-sterile plants. The time period from pollination to fertilization for interspecific crosses was about double that following self-pollination of the annual species. However, fertilization occurred frequently following interspecific pollinations. Much of the sterility was due to factor(s) operating after fertilization.Cooperative investigations of the Kansas Agricultural Experiment Station, Dept. of Agronomy and the U.S. Department of Agriculture, Agricultural Research Service. Contribution No. 82-582-J- Mention of a trademark name or proprietary product does not constitute a guarantee or warranty by the U.S. Department of Agriculture, and does not imply its approval over other products that also may be suitable.     

Use of haploids of Beta vulgaris L. for the study of orcein and giemsa stained chromosomes

Summary Some haploids isolated among the progeny from crosses involving several genotypically different motherplants of Beta vulgaris L., were used for an investigation on chromosome morphology of the genus Beta.The length of both the long and short arms proved to vary considerably for each chromosome, so that a representative idiogram of the 9 chromosomes could not be made. Staining the chromosomes according to the BSG-technique revealed only bands in the centromere regions. However, one chromosome with a narrow band in the short arm could be identified, which was supposed to be the nucleolar chromosome. The C-bands presumably correlate with the heterochromatic blocks as observed in mitotic and meiotic (pachytene) prophase.This study was part of a project on cytogenetics of interspecific hybrids resistent to beet cyst eelworm, supported by the Institute of Rational Sugar Production,Bergen op Zoom, the Netherlands.     

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.     

Crossability between some Pelargonium species

Summary Crosses were made between Pelargonium zonale, P.inquinans, P.scandans and P. stenopetalum and any interspecific hybrids obtained were selfed. The species were also selfed. Pelargonium stenopetalum was self-incompatible and cross-incompatible as the pistillate parent with the other species. As the staminate parent, P.stenopetalum was also relatively unsuccessful in interspecific crosses. Crosses between P.zonale and P.inquinans produced relatively high numbers of viable seeds and the hybrids were quite self-fertile. As indicated by the ease of crossability and hybrid self-fertility, reproductive barriers were strong between P.scandens and the other species.     

Interspecific hybridization between rice bean (Vigna umbellata) and its wild relative (V. minima): Fertility-sterility relationships

Summary The pre-and post-fertilization barriers in the interspecific crosses between Vigna umbellata and V. minima were investigated. In the reciprocal crosses (V. minima as the parent) the entry of pollen tubes into the ovary was delayed by about 4 h, and no seed set was observed. However, no pre-fertilization barriers were encountered in crosses involving V. minima as the parent and V. umbellata as the parent (normal cross). The delay/absence of divisions in the endosperm and the failure of embryo to divide were the post-fertilization barriers responsible for somatoplastic sterility in normal crosses which yielded a few hybrid seeds. The hybrid seeds showed poor germinability. The F₁ hybrids were intermediate between the parents in most morphological characters, and are completely sterile for pollen. Backcrossing of F₁ hybrid with both the parents did not restore fertility in the progenies. V. minima is considered as the tertiary gene pool of the rice bean.