The Diploidised Meiosis of Tetraploid Beta macrocarpa and its Possible Application in Breeding Sugar Beet

Accessions of Beta macrocarpa Guss., collected on the Canary Islands, were tetraploid (2n = 36). Three of these accessions were studied in detail. The plants were rather uniform in their morphological appearance, both within and between accessions, and were annual and fully self-compatible. Meiosis was completely diploidised, suggesting an alloploid nature of the tetraploid species. Crosses with diploid B. vulgaris yielded triploid hybrids which were sterile or nearly so; a few descendants of such hybrids were highly aneuploid. Crosses between tetraploid B. macrocarpa and autotetraploid cytotypes of B. vulgaris showed variable results, only part of these crosses yielded tetraploid hybrids. The tetraploid hybrids exhibited somewhat higher fertility than the triploids. An F₂ generation showed partial hybrid dwarfness, partial fertility and segregation for carliness and coloration of the hypocotyl. All hybrids had multivalents at meiosis (averages: 2.5—5.4 multivalents per pollen mother cell), indicating suppression of the diploidised meiosis. The possibilities of application of the diploidised meiosis in breeding sugar beet are discussed.     

Progenies of allotriploids of Oriental × Asiatic lilies (Lilium) examined by GISH analysis

With the aim of utilizing allotriploid (2n = 3x = 36) lily hybrids (Lilium) in introgression breeding, different types of crosses were made. First, using diploid Asiatic lilies (2n = 2x = 24), reciprocal crosses (3x − 2x and 2x − 3x) were made with allotriploid hybrids (AOA) obtained by backcrosses of F₁ Oriental × Asiatic hybrids (OA) to Asiatic cultivars (A). Secondly, the AOA allotriploids were crossed with allotetraploid (OAOA, 2n = 4x = 48), in 3x − 4x combination. Finally, the AOA allotriploids where crossed to 2n gamete producer F₁ OA hybrids (3x − 2x (2n)). Two types of triploids were used as parents in the different types of crosses, derived from: (a) mitotic polyploidization and (b) sexual polyploidization. Ploidy level of the progeny was determined by estimating the DNA values through flowcytometry as well as chromosome counting. The aneuploid progeny plants from 3x − 2x and reciprocal crosses had approximate diploid levels and in 3x − 4x crosses and 3x − 2x (2n) the progeny had approximate tetraploid levels. Balanced euploid gametes (x, 2x and 3x) were formed in the AOA genotypes. Recombinant chromosomes were found in the progenies of all crosses, except in the case of 2x − 3x crosses through genomic in situ hybridization (GISH) analyses. Recombinant chromosomes occurred in the F₁ OA hybrid when the triploid AOA hybrid was derived through sexual polyploidization, but not through mitotic polyploidization with two exceptions. Those recombinant chromosomes were transmitted to the progenies in variable frequencies.     

Resistance to Heterodera schachtii in patellares section of the genus Beta

Summary Fifty-two accessions of the section Patellares wild beet (including 26 Beta patellaris Moq., 13 B. procumbens Chr. Sm and 13 B. webbiana Moq.) and 14 progeny families were selected and tested against sugarbeet cyst nematode, Heterodera schachtii Schm. All Patellares species tested were highly resistant, but not immune, to the development of H. schachtii, after infection. This is the first report that mature female nematodes developed in the roots of B. webbiana plants. The occasional development of nematode cysts in roots of juvenile wild beets was, however, not a heritable genetic factor.Mention of a trademark, vendor, or proprietary product does not constitute a guarantee or warranty of the product by the USDA and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

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.     

Electrophoretic Investigations on Nematode Resistant Sugar Beets

Hybridizations were made between beta vulgaris and three wild species of the patellares section being resistant to the best cyst nematode {heterodera schachtii). Monosomic addition lines (2 n = 19) with full nematode resistance were investigated together with wild beets by means of electrophoretical techniques. One alkaline esterase band and a complex of several acidic esterase bands were localized on the resistance-carrying B. procumbens chromosome. The alkaline esterase marker also appeared in B. patellaris addition lines. An aconitase double band was visible in two of four B. webbiana addition lines. One resistant monotelosomic addition line with a small B. procumbens fragment had lost the esterase gene. Evidence is given that more than one chromosome is carrying genes for nematode resistance. The use of electrophoretic screening together with a nematode testing program is discussed.     

Assessment of the degree of genetic variation in beet based on RFLP analysis and the taxonomy of Beta

Summary Clones derived from Beta vulgaris and Beta maritima were assayed for their ability to detect restriction fragment length polymorphisms (RFLPs) in different beet accessions. The clones able to detect polymorphism were used as genetic markers to assess the degree of genetic variation existing between and within different species of the genus Beta. The data support the current taxonomy of the Beta vulgaris section, while the great genetic similarity found between Beta webbiana and Beta procumbens indicates that they could belong to the same species.Enough variation was found between Beta vulgaris cultivars, allowing the isolation of a sufficient number of genetic markers for the construction of detailed genetic maps.     

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.     

Evaluation of resistance to verticillium wilt in diploid,wild potato interspecific hybrids

Summary Verticillium wilt (V. albo-atrum Reinke & Berthold or V. dahliae Kleb) threatens potato (Solanum tuberosum L.) production in most growing areas of the world. Genetic resistance offers the most cost-effective and environmentally-sound control measure. However, there is a dearth of genetic and breeding information on resistance to verticillium wilt in potato, because of obscure parentage of some standard cultivars and the complex segregation at the tetraploid level. The wide range of genetic variability in wild relatives of potatoes can be potentially useful as a source of disease resistance, such as verticillium wilt resistance. Six diploid, wild, interspecific Solanum hybrids involving resistant x resistant and susceptible x resistant crosses, were assayed for verticillium wilt resistance under greenhouse conditions to evaluate their potential as sources of verticillium wilt resistance. The cross between S. gourlayi Oka. and S. chacoense Bitt. and its reciprocal had the most resistant progenies based on mean colony counts. No simple mode of inheritance can be proposed based on the observed segregation ratios. However, the continuous distributions observed on verticillium wilt disease response among hybrid families indicate that inheritance of resistance may be polygenic and complex. In addition, skewness of colony count distributions toward the resistance parents were characteristic of all resistant x susceptible crosses suggesting that resistance may be dominant. By contrast, the susceptible x susceptible cross showed a more normal distribution. Overall, the cross between S. gourlayi and S. chacoense showed the most potential as a source of verticillium wilt resistance.     

Crosses between Beta vulgaris L. and Beta lomatogona F. et M.

Summary A population of 2x^ms sugar beets was crossed with 4x Beta lomatogona F. et M. The 3x F₁-plants were male sterile and were backcrossed with 2x and 4x sugar beets and multiplied without pollination as well. After the 1st backcross mainly 3x apomict types arose again and, among others, a small number of successful 4x backcrosses. After pollination by 4x sugar beets this 4x F₁ B₁ produced. besides predominatly apomictically multiplied 4x plants, also about 7% haploid 2x hybrids. The latter probably possess 1 genome from B. vulgaris and 1 genome from B. lomatogona. In the meiosis of the PMC's a certain amount of homeology between a number of chromosomes of both species could be established. The amphihaploid hybrids can be used as breeding parents for the creation of types in which introgession can occur. During hybridization in addition to 2x and 4x B. vulgaris types a number of 2x-, 3x-, 5x- and 6x-hybrids arose. This is presumably caused by the presence of gametes with the somatic number of chromosomes and the occurrence sometimes of haploid apomictic multiplication.The presence of large numbers of bolters in the F₁ and F₁ B₁ suggests that the bolting tendency of both species is based on different genes.     

New interspecific hybrids in the genus Agropyron

Summary Fifteen species from the genus Agropyron were crossed together. Fourteen of the crosses did not produce fruits. Twenty crosses produced varying numbers of caryopses. Of the fruits produced, 22% contained no embryos. For a variety of reasons it was possible to obtain only 2 plants from the remaining 140 embryos. The crosses which yielded viable plants were between A. trachycaulum cv. Primar (2n=28) and A. intermedium cv. Chief (2n=42) as well as between A. trachycaulum from Lethbridge (2n=28) and A. desertorum cv. Nordan (2n=28). The somatic chromosome numbers for the hybrids are 37 and 28. respectively. Studies of vegetative plant character are presented.     

Successful hybridization of non-tuberous Solanum etuberosum Lind. and tuberbearing S. pinnatisectum Dun.

The results of extensive crosses between the non-tuberous species Solanum brevidens and S. etuberosum on the one hand and ten tuber-bearing Solanum species on the other are presented. Three crosses gave rise to viable progeny. Two progenies consisted of diploid plants only of the strictly self-incompatible species of the mother parent. One cross, viz. S. etuberosum × S. pinnatisectum, produced highly vigorous but fully male sterile F₁ hybrids.It is suggested that this hybrid together with those between the tomato, Lycopersicon esculentum, and S, pennellii and S. lycopersicoides constitute piers of a bridge between tomato and potato species which in the future might enable gene transfer between these two crops via their wild relatives. However, such idea has to be treated with all proper reserve.The production of this new hybrid is the first step in making accessible to potato breeding the valuable genes which have been detected in S. brevidens and S. etuberosum, viz. the genes for high resistance to frost, leafroll and Y-virus.     

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.     

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.     

Variation in some agronomically important characters in a germplasm collection of beet (Beta vulgaris L.)

Summary A geographically representative selection of germplasm of Beta vulgaris, section Beta has been assessed for characteristics important in sugarbeet breeding, including downy mildew resistance, resistance to aphid colonisation and infection by the beet virus yellow complex. The occurrence of maintainer lines for cytoplasmic male-steriles was also investigated. Desirable qualities were found in some accessions, including nothern European wild vulgaris ssp. maritima and some old multigerm cultivars of fodder beets.     

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.     

Inheritance of early blight resistance in diploid potatoes

Early blight (Alternatia solani) is a fungal disease in hot and humid environments, which causes leaf, stem and tuber lesions. Early blight resistance should be incorporated into potato cultivars because the fungicide spraying is an expensive solution for developing countries. The diploid cultivated species Solanum tuberosum group Phureja and group Stenotomum are sources of resistance alleles. The elucidation of the inheritance for early blight resistance must help to decide what could be the best breeding procedure to improve this diploid germplasm and transfer the resistance to the tetraploid level. Three experiments were carried out under controlled and field conditions to determine the heritability of this trait using nested and diallel mating designs with haploid, species and haploid-species hybrids. The narrow-sense heritability estimates were relatively high (0.64–0.78). This means that additivity was the most important type of gene action for determining resistance to early blight at the diploid level. The results suggested that diploid parents showing highest levels of resistance, throughout the cycle of disease development, can be used in 4x×2x crosses to obtain resistant tetraploid progenies to this fungal disease.     

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.     

Interspecific hybrids of Cyclamen persicum Mill. and C. purpurascens Mill. produced by ovule culture

Summary Interspecific crosses were made to introduce the scent of flowers of C. purpurascens into C. persicum cultivars and ovule culture was used to rescue the abortive hybrid embryos. Cultivars of C. persicum diploid (CPD, 2n=2×=48) and C. persicum tetraploid (CPT, 2n=4×=96) were the pistillate parents and wild species of C. purpurascens (CP, 2n=34) were staminate parents. After pollination, crossed ovaries were collected periodically and examined using paraffin sections. Histological observations suggested that both hybrid ovules of CPD x CP and CPT x CP should be transferred to culture medium 35 days after pollination. Based up on this observation, crossed ovaries were collected 28 days after pollination and ovules with placenta were transferred to MS (1962) medium containing 3% sucrose. These ovules were cultured in the dark at 25° C. The hybrids (2n=41) derived from CPD x CP had the scent of C. purpurascens, whereas the hybrids (2n=65) derived from CPT x CP had the scent of C. persicum. Although both hybrids had complete genomes from the parents and produced a few viable pollen grains, they failed to yield viable seeds by self- and cross-pollination with fertile pollen grains of C. persicum cultivars.Abbreviations CPD C. persicum diploid - CPT C. persicum tetraploid - CP C. purpurascens     

Study on lily introgression breeding using allotriploids as maternal parents in interploid hybridizations

Based on a recent hypothesis, “Five same genomes of endosperm are essential for its development in Lilium”, it is expected that allotriploid lily (OTO) can be hybridized with diploid Oriental lily (OO) for introgression breeding in Lilium L.. To test the hypothesis, OTO lilies, ‘Belladonna’, ‘Candy Club’ and ‘Travatore’, were used as the maternal parents and crossed with two diploid OO cultivars, ‘Siberia’ and ‘Sorbonne’, and the species L. regale Wilson (TT). Results showed that capsules of all OTO × OO hybridizations developed well and 0.8~3.3 viable seedlings per ovary were obtained through normal pollination and embryo rescue; however, all OTO × TT crosses failed. Genomic in situ hybridization showed that the progenies of the OTO × OO hybridizations were aneuploid and a variable number of T-genome chromosomes were introduced into the progenies through the allotriploid lilies. The present results not only demonstrate that allotriploid OTO lilies, although male sterile, can be used as maternal parents to produce aneuploid progenies, but also strongly support the new hypothesis in lily breeding.     

Observations of fruit shape and other characters in the species of the section Patellares,genus Beta

Summary Doubt is cast on the classification of the species within the section. Although Beta patellaris is distinct, the complete interfertility and intergradation of morphological characters between B. webbiana and B. procumbens suggests them to be two extremes of a single ecospecies. This has significance in studies of hybridization between sugar beet (B. vulgaris) and the members of this section.