Double-bridge hybrids of Solanum bulbocastanum and cultivars of Solanum tuberosum

Summary Solanum bulbocastanum (2n=2x=24) has valuable characters for potato breeding, but cannot be hybridized directly with S. tuberosum cultivars. Both S. acaule (2n=4x) and S. phureja (2n=2x) were used as bridging species. Triploid S. acaule × S. bulbocastanum were doubled with colchicine and the resulting fertile hexaploid F₁'s crossed with S. phureja. The triple hybrids obtained were tetraploid or nearly so. The two genomes of S. acaule in these triple hybrids probably pair preferentially, which may provoke pairing and possibly crossing over between the chromosomes of S. bulbocastanum and S. phureja.More than 20000 pollinations of the triple hybrids with four potato cultivars had to be made to produce 40 quadruple hybrids. These highly vigorous hybrids varied greatly in many morphological characters, resistance to Phytophthora infestans, fertility and crossability. The chromosome numbers are 48 (24 hybrids), 49 and 46, but some higher ploidy levels (65, 66, 72 chromosomes) were found as well. Their origin is to be sought in the fusion of an unreduced egg cell from triple hybrids (either euploid or hypoploid) and a reduced male gamete from the cultivars. This view is corroborated by their extreme resistance to Phytophthora. Also some 48-chromosome hybrids are highly resistant, which may indicate introgression from S. bulbocastanum.Most quadruple hybrids are readily inter-crossable and crossable as females with cultivars; several also as males. Two could be hybridized with S. bulbocastanum, but the few seeds dit not germinate.Studies of pachytene stage of meiosis revealed the presence of a S. bulbocastanum chromosome in at least one tetraploid hybrid, which is highly resistant to Phytophthora. At metaphase I of meiosis chromosome associations higher than quadrivalents were not found. Except in one hybrid, the frequency of quadrivalents did not exceed one per cell and the average proportion of chromosomes associated as bivalents amounted to 90%.The quadruple hybrids (double-bridge hybrids) appear good starting material for breeding programmes aimed at introducing genes from S. bulbocastanum into S. tuberosum cultivars.     

Fertile somatic hybrids of Solanum etuberosum (+) dihaploid Solanum tuberosum and their backcrossing progenies: relationships of genome dosage with tuber development and resistance to potato virus Y

The wild non-tuberous species Solanumetuberosum is resistant to biotic andabiotic stresses, but is very difficult tocross with cultivated potato. Therefore,interspecific somatic hybrids between adihaploid clone of potato S.tuberosum (2n=2x=24, AA genome) and thediploid species S. etuberosum(2n=2x=24, EE genome) were produced byprotoplast fusion. Among the 7 fertilefusion hybrids analysed by genomic insitu hybridisation (GISH), three groups ofplants were found with the genomicconstitution of AAEE, AAEEEE and AAAAEE.Four fusion hybrids had exactly theexpected chromosome composition, while eachof the three aneuploid hybrids had lost twochromosomes of S. etuberosum. Twobackcross progenies were developed, andGISH analysis was applied to analysetransmission of the parental chromosomesinto the sexual generations. BC₁hybrids derived from the crosses of thehexaploid somatic hybrids with tetraploidpotato were pentaploid with thetheoretically expected genomic compositionor with slight deviation from thisexpectation. In the three BC₂ hybridsanalysed by GISH seven to 12 chromosomes ofS. etuberosum were detected in thepredominant S. tuberosum background.No recombinant chromosomes in the hybridswere detected. Genome dosage affects tuberformation in hybrids and their progenies,but has less effect on resistance to potatovirus Y (PVY) in fusion hybrids. Severalgenotypes of the fusion hybrids andBC₁ progeny did not show viralinfection even in the graftingexperiments.     

Tuber yield and quality of 4x-2x (FDR) potato progenies derived from the wild diploid species Solanum berthaultii and Solanum tarijense

Field experiments were carried out in order to evaluate 4x‐2x families derived from crosses between elite 4x potato cultivars and 2x Tuberosum‐Solanum tarijense and 2x Tuberosum‐Solanum berthaultii clones. Three traits were assessed: total tuber yield (TTY), haulm maturity (HM) and general tuber appearance (GTA). The degree of heterosis of these hybrid families was evaluated by comparison with the respective 2x and 4x parents as well as with seven 4x cultivars. The parental haploid species hybrids derived from S. berthaultii and S. tarijense combined two or more positive horticultural characteristics. Expressed as yield percentage of the 4x parents, the TTY of the families ranged from 53% to 246%. For TTY, the best 4x‐2x hybrid family ranked better than seven out of nine elite 4x cultivars. Some families had GTA scores in the range of the highly selected 4x cultivars. The families, however, were generally later maturing than the 4x parent group. Specific combining ability for TTY and GTA were the only two significant sources of variation observed in this genetic material. Parent‐offspring correlation coefficients were low for all traits, and indicated that parental performance would not be informative at either ploidy level. These results parallel previous investigations with distinct haploid species hybrids where a 4x‐2x breeding scheme was found to be an effective strategy for increasing progeny TTY over the 4x parents. However, the high degree of heterosis for TTY along with good GTA scores observed in certain cross combinations derived from these unadapted 2x species was a somewhat surprising result. Thus far, the importance of these two South American wild potato species, from the potato‐breeding standpoint, has been limited to the fact that they are natural reservoirs of major genes controlling resistance against important diseases and insects. However, the level of performance of some 4x‐2x families in comparative assays with elite cultivars suggests the unanticipated possibility of introgressing genetic factors from S. berthaultii and S. tarijense with positive effects on quantitative traits of horticultural importance along with these major resistance genes.     

Alteration of the genomic composition of Solanum nigrum (+) potato backcross derivatives by somatic hybridization: selection of fusion hybrids by DNA measurements and GISH

Fusion experiments were performed with a first (BC₁‐6738) and a second (BC₂‐9017) generation backcross hybrid of 6x Solarium nigrum (+) 2x potato somatic hybrids with potato cultivars. Because no progeny was obtained from the BC₂ genotypes, alternative approaches were sought to overcome the sexual crossing barrier. Five potato genotypes, one of which contains the hygromycin resistance gene, were used in the fusion experiments. All vigorous regenerants were used for the estimation of nuclear DNA content using flow cytometry. Plants with a DNA content higher than that of the BC₁‐6738 or BC₂ genotypes were considered potential somatic hybrids. Forty‐nine potential somatic hybrids resulted from fusion experiments with BC₁‐6738, from which 20 grew vigorously in the greenhouse and flowered. After pollination with several 4x potato cultivars, eight genotypes produced seeded berries and five genotypes gave seedless berries. In addition, 11 of these 13 somatic hybrids were selected for genomic in situ hybridization (GISH) analysis to determine their genomic composition. Nine had exactly or approximately the expected number of 36 S. nigrum and 60 potato chromosomes. In one genotype, only 22 instead of 36 S. nigrum chromosomes were found and one potato chromosome was possibly missing. Only five potential somatic hybrids were detected among the 79 regenerants from BC₂‐9017 (+) 2x potato fusion experiments that were analysed by flow cytometry. Two of these hybrids were rather vigorous and did flower, but pollinations with potato have not yet set any berries.     

Somatic hybridization between Solanum tuberosum and species of the S. nigrum complex: Selection of vigorously growing and flowering plants

Fusion experiments were performed between diploid (2n = 2x = 24) or tetraploid (2n = 4x = 48) potato genotypes and four species of the Solanum nigrum complex, namely S. nigrum (2n = 6x = 72), S. villosum (2n = 4x = 48), S. chenopodioides (2n = 2x = 24) or S. americanum (2n = 2x = 24 and 2n = 6x = 72). All five accessions of the S. nigrum-species were successfully hybridized with at least one of the potato genotypes. Somatic combining abilities were influenced by the ploidy level as well as the genotype of the parental species. The use of kanamycin or hygromycin resistance as cell-selectable markersystem had no influence on somatic combining ability, but such markers can be useful to improve efficient selection of somatic hybrids in sufficient numbers. At least 20% of the hybrids of each successful combination performed well in vitro. However, only 60 genotypes out of 761 somatic hybrids were vigorous as well as flowering in the greenhouse. Analysis of the DNA content of somatic hybrids could be used as a criterion for the indirect selection in vitro of hybrids that were vigorous in the greenhouse. Flowering somatic hybrids of S. nigrum (+) 2x potato and S. americanum (+) 4x potato were selected with the aim of introgression of resistance traits after recurrent backcrossing with cultivated potato. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genome composition of triploid lily cultivars derived from sexual polyploidization of Longiflorum × Asiatic hybrids (<Emphasis Type="Italic">Lilium</Emphasis>)

About 19 cultivars, which had originated from backcrosses between F1 LA (Longiflorum × Asiatic) hybrids (2n = 2x = 24) as female parents and Asiatic cultivars as male parents (2n = 2x = 24), were analyzed with genomic in situ hybridization. 17 of them were triploid (2n = 3x = 36), and two aneuploid (2n = 3x + 1 = 37). The triploid cultivars had resulted from the functional 2n eggs produced by the female parents (F1 hybrids) because first division restitution (FDR) occurred in their meiosis during megasporogenesis. Similarly, the aneuploid cultivars had originated from viable 2n + 1 eggs. The extra chromosome in cultivar 041555 or 041572 resulted from one univalent or one half-bivalent which might have lagged behind when the sister chromatids of the other univalents and half-bivalents were segregating during the FDR process in their LA hybrid parents, respectively. That the majority of cultivars possessed recombinant chromosomes showed that intergenomic recombination might play an important role during the selection of the cultivars directly from BC1 progenies. That five cultivars of the 15 recombinant cultivars only had reciprocal recombinant chromosomes and 10 cultivars had non-reciprocal recombinant chromosomes indicates that the latter are more important. Because 9 of the 10 non-reciprocal recombinant cultivars possessed substitutions for recombinant segments, it also indicated that such substitutions could be an important source for the genetic variation in the sexual triploid BC1 progenies. In such cases there was a potential for the expression of the recessive genes of the backcross parent in a nulliplex (aaa) condition in the substituted segments. Genetic variation resulting from such nulliplex loci might have played a role in the selection of some of the cultivars.     

Evolutionary autoploidy in the sweet potato (Ipomoea batatas (L.) Lam.) and its progenitors

Summary The role of polyploidy in the evolution of the sweet potato. I. batatas (2n=6x=90), became more clear in 1971 when wild species with 30, 60, and 90 chromosomes were discovered. These species, I. leucantha (2x), I. littoralis (4x) and I. trifida (6x), are the progenitors of the sweet potato (6x), in an autopolyploid series with doubling of the I. leucantha B genome.In the present study the hypothesis of the origin of the sweet potato was confirmed by comparative studies on some plant characters, sexual compatibility, and the behavior of artificial hexaploids produced from I. leucantha (2x) and I. littoralis (4x).Since induced hexaploid I. leucantha exhibits weak growth, a few multivalents are formed at meiosis in the sweet potato, and there are differences in morphological and physiological characteristics between the artificial hexaploids and I. batatas, raw autoploidy seems unlikely, but a balanced or diploidised autoploidy could have been achieved by genic and chromosomal changes in the course of evolution. However, there is still sufficient homology so that meiotic pairing occurs usually between regular partners but also between homoeologous chromosomes in a certain situation.     

Induction of haploids and aneuhaploids in colchicine-induced tetraploid Solanum chacoense Bitt.

Summary The species Solanum chacoense BITT. (2n=2x=24) is a tuber-bearing, self-incompatible species which is important both for breeding and for genetic research. It crosses readily with most other tuber-bearing Solanum species including the common potato S. tuberosum (2n=4x=48). Gametophytic incompatibility hampers research in and utilization of this species. Doubling the chromosome number by colchicine makes it a self-compatible autotetraploid. By crossing selfed progeny of 4x-S. chacoense with a number of haploid-inducing diploid Solanum species a high yield of different dihaploid S. chacoense individuals (2n=24) could be obtained from one originally diploid clone: S. chacoense CPC 1153. More than 160 haploids showing a large variability were identified. The average haploid frequency was 53.7 per 100 berries. Most hybrid plants (70–100%) from four 4x × 2x crosses studied were tetraploid. The frequency of triploid hybrids was low (0–10%). Haploid-inducing capacity of fifteen male parents used in this study varied from 0–141 haploids per 100 berries. Careful examination of 156 haploids revealed 15 viable aneuhaploids (2n=25, 26, and 27), i.e. 9.4%. The potential value of these aneuhaploids is discussed.About 50% of the haploids were sufficiently male fertile to use them in crossing. A few of them set berries after selfing. Five aneuhaploids including the one with 27 chromosomes were successfully crossed as females with a diploid hybrid clone.     

Breeding behavior for tuber protein in Solanum tuberosum and Tuberosum-phureja hybrids

Summary True protein content among tubers within a plant of Solanum tuberosum cv. Oneida was found to be negatively correlated with tuber size. A similar study of S. phureja genotype 148-17 revealed no correlation. Tuber protein was determined for 250 genotypes in each of four hybrid potato populations in a factorial mating design with four potato cultivars as stylar parents mated to four groups of pollen parents (4x cultivars, 4x high protein selections, 2x S. phureja, 2x high protein S. phureja). The hybrid population derived from the 4x high protein selections was significantly higher in protein content than the other populations. High estimates of general combining ability for tuber protein content were found for both stylar and pollen parents. Poor photoperiodic adaptation to growing conditions in northern latitudes was a possible explanation for the variable protein phenotypes typical of S. phureja and the lack of transmission of the high protein character in phureja to 4x-2x hybrids. Although protein content was negatively correlated with total yield, high protein segregates with good yield potential were identified in all four populations.Scientific Journal Series Article 11, 616 of the Minnesota Agricultural Experiment Station.     

Male fertility and cytology of triploid hybrids between tetraploidSolanum commersonii (2n=4x=48, 2EBN) and Phureja-Tuberosum haploid hybrids (2n=2x=24, 2EBN)

Summary Solanum commersonii Dun. is a diploid (2n=2x=24, 1EBN) wild species of potential value for potato breeding. It is a reproductively isolated species and cannot be crossed with Tuberosum haploids (2n=2x=24, 2EBN) or other diploid 2EBNSolanum species. In order to overcome the EBN barriers, triploid hybrids were produced between Phureja-Tuberosum haploid hybrids, which form 2n pollen grains by parallel spindles, and tetraploidS. commersonii. Microsporogenesis analysis of the triploids indicated a trend towards low values of chromosome distribution at Anaphase I; lagging chromosomes were often observed as well. Despite these abnormalities, the percentage of stainable pollen was very high, ranging from 5.0% to 74.3%. A high variation in pollen grain diameter was also evident. Parallel and tripolar orientation of spindles at Metaphase II of microsporogenesis was a common feature of all the triploids analyzed, but dyads and triads were observed at a very low frequency. Therefore, also the frequency of 2n pollen was very low; the different size of stainable pollen appears to represent the ploidy levels which are possible according to the distribution of chromosomes in Anaphase I. The results obtained also suggest thatS. commersonii could have minor genes acting at the end of meiosis in such a way that, despite the presence of parallel/tripolar spindles, dyads/triads are not formed.Contribution no. 124 from the Research Center for Vegetable Breeding.     

The effect of colchicine treatment on Solanum acaule and S. bulbocastanum; A complete analysis of ploidy chimeras in S. bulbocastanum

Summary Seeds of tetraploid Solanum acaule (2n=48) and diploid S. bulbocastanum (2n=24) were germinated in petri-dishes on filter paper soaked in 0.3% colchicine. An additional treatment with 0.3% colchicine was applied one month after sowing at four successive days in the axils of the cotyledons of the seedlings. S. acaule appeared much more sensitive to colchicine (14 surviving seedlings from 500 seeds) than S. bulbocastanum (109 surviving seedlings from 450 seeds). Six S. acaule plants with 2n=96 chromosomes were obtained against 38 S. bulbocastanum plants with 2n=48 chromosomes.The ploidy level in each of the three germ layers L₁, L₂, L₃ was determined in 113 plants of S. bulbocastanum and the following results were obtained. Four of the eight possible ploidy types were detected, viz 2x-2x-2x (72 plants), 4x-2x-2x (3 plants), 2x-4x-4x (9 plants) and 4x-4x-4x (29 plants). Doubling the number of chromosomes resulted in a highly significant increase of the number of chloroplasts in the guard cells of stomata and a greatly significant decrease in the proportion of trimerous pollen, male fertility and leaf index. The variability for all characters studied, except for leaf index, was clearly lowest in the 2x-2x-2x group. All plants with a 2x-L₂ were highly male fertile and self-incompatible, also in the three bud stages tested. Male fertility of the plants with 4x-L₂ varied greatly: 12 plants had more than 90% stainability, 5 plants must be considered male sterile. All non-sterile plants with 4x-L₂ were found to be self-compatible, pointing to a gametophytic system of incompatibility in S. bulbocastanum.     

Assessing success in gene transfer betweenLolium multiflorum andFestuca arundinacea

Summary AFestuca arundinacea (2n=6x-42) plant with three PGI/2 homoeoalleles marking three homoeologous chromosomes was crossed with aLolium multiflorum plant (2n=4x=28) with a different PGI/2 phenotype to give a pentaploid hybrid (2n=5x=35) with five chromosomes each marked by a different PGI/2 allele. This hybrid plant was backcrossed twice with diploidL. multiflorum (2n=2x=14) with a different PGI/2 phenotype. Numbers of interspecific recombinants involving chromosomes marked by PGI/2 were then determined in both backcross generations. In the BC1, recombinants involving only one PGI/2 allele were found but in the BC2, all three homoeologousF. arundinacea chromosomes carrying the PGI/2 locus recombined withLolium with one in greater frequency and the others in equal but lower frequency. The evidence supports claims made forF pratensis (2n=2x=14) andF. arundinacea var.glaucescens (2n=4x=28) being progenitors forF. arundinacea.     

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.     

Karyotype analysis and interspecific hybridisation in three perennial Sesbania species (Leguminosae)

Summary The somatic chromosome number in Sesbania sesban var. nubica, S. goetzei and S. keniensis (Leguminosae; Papilionoidae) was found to be 2n=12. These findings were in agreement with earlier reports on S. sesban and S. keniensis. The chromosome number 2n=12 is a new record for S. goetzei. Similarities in karyotypes were found in the three species. All species had one pair of long metacentric chromosomes; the second pair was submedian, followed by four smaller pairs of metacentric chromosomes. Nucleolar organiser regions in the form of satellites were found on the short arm of the fourth chromosome pair in S. sesban and S. keniensis. Interspecific crosses in all possible combinations were carried out, resulting in pod and viable seed formation for the crosses S. sesban x S. goetzei, S. sesban x S. keniensis, S. goetzei x S. sesban and S. goetzei x S. keniensis. The two crosses with S. keniensis as a female parent were unsuccessful. The hybrid plants established normally and produced viable seeds.     

The transfer of the synaptic gene (sy-2) from 1EBN Solanum commersonii Dun. to 2EBN germplasm

Summary Crosses of a synaptic mutant (sy-2) of 2x(1EBN) S. commersonii Dun. with 2x(2EBN) S. chacoense Bitt. resulted in the production of 3x(2EBN) hybrids. Low levels of 2n egg production in these hybrids allowed subsequent crosses to be made with 4x(4EBN) cultivars, resulting in 5x(4EBN) S. commersonii-S. chacoense-Group Tuberosum hybrids. Seed set in these crosses averaged 0.22 seeds per fruit. Fifty-four unspotted seeds were produced in 1225 crosses of the 5x(4EBN) hybrids with 2x(2EBN) Group Phureja haploid extraction clones carrying dominant seed spot markers. These seeds yielded a total of 15 confirmed haploids having a diploid (2n=2x=24) or near diploid chromosome number. One haploid clone recovered displayed a synaptically abnormal phenotype similar to the original S. commersonii synaptic mutant. Preliminary test crosses of these haploids indicated that all haploids produced were 2EBN.This method for the transfer of alleles across EBN levels holds promise for the recovery of both qualitative and quantitative traits from 1EBN species. In these recovered, cross-compatible forms, clones with desired characteristics may be selected and easily incorporated into breeding programs.     

The Use of Potato Haploids to Put 2x Wild Species Germplasm into a Usable Form

An effective method of incorporating germplasm of wild 2x Solanum species into the cultivated potato is through hybridization with haploids of 4x S. tuberosum Group Tuberosum. This procedure provides for the introduction of desirable traits and genetic diversity from wild species into potato. Haploid and wild species genotypes can be evaluated for parental value based on the presence of good tuberization, desirable traits, fertility, and 2n gametes in their hybrid progeny. Haploid-wild species hybrids may be intercrossed and evaluated at the 2x level to take advantage of diploid genetic ratios. Selected 2x genotypes may then be brought to the 4x level through sexual polyploidization with 2n gametes.     

DNA analysis of potato + Solanum brevidens somatic hybrid lines

Three somatic hybrid lines between potato (cv. While Lady line no. Ke 79, 2n = 2x = 48) + Solanum brevidens (PI 218228, 2n = 2x = 24) were evaluated using randomly amplified polymorphic DNA (RAPD) markers. The lines originated from the same callus but showed different reactions to Erwinia carotovora ssp. carotovora, the cause of potato soft rot. By the use of 48 oligomer primers producing 99 scorable bands, DNA polymorphism were detected on 7 of 12 S. brevidens chromosomes. Loss of certain DNA segments on chromosome 5, 6, 9 and 11 were observed. Some of the variations could have taken place in early callus stage of development; others may have occurred after initiation of individual shoot regeneration. The possible involvement of missing RAPD products specific to one somatic hybrid that shows decreased resistance to bacterial soft rot is discussed.     

Amphidiploids between Cyclamen persicum Mill. and C. purpurascens Mill. induced by treating ovules with colchicine in vitro and sesquidiploids between the amphidiploid and the parental species induced by conventional crosses

Summary We cultured colchicine-treated hybrid ovules in vitro to produce fertile amphidiploids of C. persicum (2n=2x=48. referred to as AA) × C. purpurascens (2n=2x=34, referred to as BB). Seedlings and mature plants were obtained from the ovules without colchicine and those exposed to 50 mg/l colchicine for 5, 10 and 15 days, whereas they were not obtained from the ovules exposed to 50 mg/l colchicine for 20 days and 500 mg/l for 5, 10, 15 and 20 days. Although 8 mature hybrids derived from the ovules without colchicine produced a few fertile pollen grains, they failed to produce viable seeds by self-fertilization. The hybrids had 41 somatic chromosomes. Four and 3 mature plants were derived from ovules exposed to 50 mg/l colchicine for 10 and 15 days, respectively. One each among 4 and 3 mature plants showed a high frequency of pollen grain fertility, produced several seeds by self-fertilization, and had 82 somatic chromosomes which is twice the number of hybrid chromosomes (2n=41, AB). These findings indicated that these plants are amphidiploids (2n=82, AABB) between C. persicum and C. purpurascens. Three and 2 viable seeds were derived by the conventional crosses of diploid C. persicum × the amphidiploid and the amphidiploid × C. purpurascens, respectively. Flowering plants that developed from the seeds of diploid C. persicum × the amphidiploid were barely fertile and had 65 somatic chromosomes (2n=65, AAB), whereas those that developed from the seeds of the amphidiploid × C. purpurascens were barely fertile and had 58 somatic chromosomes (2n=58, ABB). The somatic chromosomes indicated that these plants are probably sesquidiploids between the amphidiploid and either C. persicum or C. purpurascens. The interspecific cross-breeding of cyclamen using the amphidiploids and the sesquidiploids is discussed.     

Meiosis in different F1-hybrids of Solanum acaule Bitt. x S. bulbocastanum Dun. and its bearing on genome relationship,fertility and breeding behaviour

Summary Meiosis was studied in some triploid, tetraploid and hexaploid F₁-hybrids from Solanum acaule x S. bulbocastanum and in a triploid F₁-hybrid from S. acaule x S. tuberosum-haploid.The only anomaly found was stickiness at metaphase I, the degree of which appeared to be related to the proportion of the S. bulbocastanum chromosomes in the hybrids. No stickiness occurred at pre- and post-metaphase stages.A clear allosyndetic pairing between chromosomes of the two S. acaule genomes was observed in all triploid and tetraploid hybrids. These genomes apparently are closely related and little differentiated. The triploids S. acaule x S. bulbocastanum had 0–1 trivalent per cell, whereas 2–5 trivalents per cell were observed in the triploid S. acaule x S. tuberosum-haploid. Therefore S. acaule is more closely related to S. tuberosum L. than to S. bulbocastanum. A small amount of pairing between S. acaule and S. bulbocastanum is apparent from the occurrence of multivalents in all hybrids.Hexaploid F₁'s (2n=72) showed a nearly complete homologous pairing of chromosomes (35.2 bivalents per cell) and generally a normal separation of chromosomes at anaphase: 36-36. This offers an explanation for their high fertility. Triploid F₁'s from S. acaule x S. bulbocastanum have a high frequency of univalents leading to irregular separation of chromosomes at anaphase and consequently to unbalanced gametes and extreme sterility. In the tetraploid F₁'s (2n=48) nearly complete bivalent pairing was observed, 50% expectedly being homologous and 50% homoeologous pairing. Separation of chromosomes at anaphase was generally normal 24-24. In spite of this normal behaviour and allowing for tight stickiness at metaphase the tetraploids are very sterile. A satisfactory explanation cannot yet be given.Selfing and intercrossing hexaploid F₁'s gives normal berry set and many seeds per berry. However crosses with S. tuberosum and even those with the fertile hexaploid hybrid from 8x-S. acaule x 4x-S. tuberosum are little successful: berry set is far below normal and the berries are either parthenocarpic or contain only one or two seeds. These rather unexpected results warrant further investigation. Large-scale selfings and intercrosses of triploid and tetraploid hybrids have not been successful as yet. Among the female gametes of tetraploid hybrids a few appeared to be functional in crosses with hexaploid hybrids and in those with S. bulbocastanum.     

Molecular cloning of Tulipa fosteriana rDNA and subsequent FISH analysis yields cytogenetic organization of 5S rDNA and 45S rDNA in T. gesneriana and T. fosteriana

In this study, Tulipa fosteriana was found to contain 45S rDNA repeat units of 9.7 and 9.5 kb, in which at least 7 types of 45S rDNAs were identified by restriction site analysis. For 5S rDNA, repeat units ranging from 364 bp to 396 bp were identified. The diploid cultivars (2n = 2x = 24) ‘Christmas Dream’ and ‘Queen of Night,’ representing the horticultural group T. gesneriana, and ‘Red Emperor’, belonging to T. fosteriana, were compared cytogenetically using cloned 5S and 45S rDNAs. Fluorescence in situ hybridization (FISH) analysis identified many rDNA sites located on each chromosome in the diploid genomes. For example, we identified 71 sites of 5S rDNA and 10 sites of 45S rDNA in ‘Red Emperor’. Additionally, FISH analyses enabled construction of karyotypes for these cultivars. Karyotype comparison of T. gesneriana cultivars showed conservation of repetitive rDNA unit positioning. A clear difference in chromosome size and signal pattern was observed between T. gesneriana and T. fosteriana cultivars. Here we demonstrate the unique nature of the highly repeated 5S rDNA units in these Tulipa species and the usefulness of FISH karyotyping with cloned 5S and 45S rDNAs to clearly distinguish between chromosomes from T. gesneriana and T. fosteriana. Hitoshi Mizuochi and Agnieszka Marasek contributed equally to this paper