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.     

Bilateral polyploidization inDactylis glomerata L. subsp.lusitanica: occurrence,morphological and genetic characteristics of first polyploids

Summary InDactylis glomerata L. subsp.lusitanica, triploid and tetraploid plants were obtained by bilateral sexual polyploidization in crosses between diploid parents known to produce 2n gametes. The polyploid and diploid progeny were compared for allozyme diversity (allele number and heterozygosity), phenological (pollen fertility, inflorescence emergence date), cellular (stomatic cell size) and morphological characters (vegetative biomass, seed weight, total seed number per plant, panicle number and seed number per panicle length) in four contrasting growing conditions. Stomatal cell size and the total number of alleles were significantly lower in diploids than in polyploids which sometimes recovered more than two distinct alleles at a locus via 2n gametes possessing heterozygous loci. Pollen fertility was similar in diploids and tetraploids, but significantly lower in the triploids. Seed weight was significantly higher in polyploids, although the average number of seeds per panicle length and the total number of seeds was higher in the diploid sibs. No phenological differences occurred among ploidy levels whereas such differences were observed inDactylis in all the natural areas of sympatry between diploids and their related polyploids. Our results provide evidence for an instantaneous effect of polyploidization on certain traits and suggest that natural selection would act subsequently on several other traits to allow the establishment of new polyploids.     

Molecular marker diversity and bacterial wilt resistance in wild <Emphasis Type="Italic">Solanum commersonii</Emphasis> accessions from Uruguay

Solanum commersonii is a wild tuber-bearing species native to Uruguay with high potential for use in potato breeding programs. Little is known about the genetic diversity within this wild species and the relationship with the resistance to the bacterial pathogen Ralstonia solanacearum. We studied 30 S. commersonii clonal accessions, 20 of which were collected from geographically different areas across the country, while the other ten were grown from seeds from a single plant. Resistance against R. solanacearum was tested and different levels of resistance were found, ranging from delayed wilting to asymptomatic reactions. The genetic variation and the relationships among individuals in this germplasm collection were studied by different molecular markers: Random Amplified Polymorphic DNA (RAPD), Amplified Fragment Length Polymorphism (AFLP) and Microsatellites or Simple Sequence Repeats (SSR). AFLP markers generated the largest number of total and polymorphic fragments per assay unit while SSR revealed the highest frequency of polymorphic bands (100%), followed by AFLP (96.2%) and RAPD (89.4%). In contrast, when comparing the number of different genetic profiles generated, the SSR markers exhibited the lowest discriminatory power. The clustering pattern obtained with the three marker systems showed a similar distribution of the S. commersonii germplasm revealing a high correlation between the three methods employed. All three dendrograms grouped most of the accessions into two main clusters, containing the same accessions regardless of the marker type. Bacterial wilt resistant accessions were present in both clusters. Accessions originated from different seeds of the same plant were grouped within one of the major clusters, and differed in the response to R. solanacearum revealing segregation of resistance. Furthermore, the distribution in two main clusters showed high correspondence with the geographical origin of the accessions, from the north and south of the country, and with the subspecies malmeanum and commersonii morphologically identified.     

Breeding potential of Solanum tuberosum–S. commersonii pentaploid hybrids: fertility studies and tuber evaluation

Twenty-one (near) pentaploid hybrids between sexually incompatible Solanum commersonii [2x(1EBN)] and cultivated S. tuberosum [4x(4EBN)] were characterized for tuber traits and fertility. A number of genotypes resembled the S. tuberosum type in terms of stolon length and eye depth and produced tubers under long day conditions. Tuber yields were not as high as expected, probably due to lack of adaptation of the S. commersonii genome to the environmental conditions in southern Italy (on average 325 g·pt⁻¹ and 285 g·pt⁻¹ in 2003 and in 2004, respectively). Compared to 2004, in the summer of 2003, characterized by extremely high temperatures, hybrids gave a higher tuber yield than the S. tuberosum control, suggesting that in our environmental conditions the wild S. commersonii genome, rather than resulting in heterosis for tuber yield, provides better adaptation to harsh environments. Although aneuploidy has often been associated with reduced fertility, several hybrids were fertile in crosses with S. tuberosum when used as female parents. In particular, the average berry set and number of seeds per berry were 38.2% and 31.8%, respectively. Based on significant relationships between ploidy levels and all yield and fertility data measured, the presence of extra chromosome affected the parameters considered: tuber yield in 2003 and 2004, percentage of fruit set, number of seeds per berry and number of seeds per pollinated flower. Regression analysis also indicated that “Residuals” were significant for all parameters measured. Therefore, additional factors (e.g. the genetic makeup of hybrids) may be key to fertility and yield.     

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.     

Natural triploid hybrids between Solanum acauleBitter and S. infundibuliformePhilippi in the province of Jujuy,Argentina

Summary The occurrence of natural triploid hybrids between tetraploid Solanum acauleBitter subsp. acaule and diploid S. infundibuliformePhilippi both tuber-bearing Solanums, in the Puna region of Jujuy, is reported. The natural hybrids, which are morphologically intermediate between their putative parents and completely male sterile, have a somatic chromosome number of 36. The artificial hybrids, obtained by crossing both parental species, closely resemble the natural ones thereby confirming their parentage. It is proposed to designate the natural hybrids Solanum x viirsooi hybr. nov.     

Natural hybridization between Solanum acaule Bitt. and S. megistacrolobum Bitt. in the province of jujuy,Argentina

Summary Evidence is provided that hybridization of the tetraploid (2n=4x=48), self-fertile tuber-bearing species Solanum acaule Bitt. with the diploid (2n=2x=24), self-incompatible, tuber-bearing S. megistacrolobum Bitt. takes place in several localities of the province of Jujuy in the high, cold plateaux of the Argentine Puna. The triploid hybrids (2n=3x=36) closely resemble S. acaule in growth habit, leaf morphology and floral structures and for this reason they can be easily overlooked for that species in the field. Experimental data show that S. acaule can be crossed with S. megistacrolobum though the crossability is rather low and variable according to the particular cross considered. The artificial hybrid obtained compares well with the natural hybrid in morphology and chromosome number. The hybrids, though almost completely male sterile, are successful colonizers of disturbed areas around farmers' dwellings, cattle enclosures and other areas where the soil is rich in organic matter.There is some evidence that the Tilcara material of S. acaule subsp. aemulans and the hybrids of S. acaule x S. megistacrolobum have some characters in common, which can be interpreted as having a similar origin.It is postulated that S. acaule subsp. aemulans, in Jujuy at any rate, is not a primitive form of S. acaule as thought by Hawkes and Hjerting, but rather a fertile hybrid derivative of S. acaule x S. megistacrolobum through the functioning of 2n gametes.We also provide evidence that S. bruecheri Correll should not be considered a hybrid of S. acaulle x S. megistacrolobum but a synonym for S. gourlayi Hawkes. The new name, S. x indunii Okada et Clausen, is proposed to designate this hybrid.     

The domestication and improvement of cowpeas (Vigna unguiculata (L.) Walp.)

Summary Physiological and morphological characteristics of the two wild and three domesticated subspecies of cowpeas are compared. The wild accessions are alike in having small, hard seeds borne in dehiscent pods, but differ in other characteristics. We suggest that the wild subsp. dekindtiana, from the seasonally-arid tropics, is more likely to have been the progenitor of modern cowpeas than the other wild subspecies (subsp. mensensis), but that subsp. dekindtiana was first cultivated in the humid tropics where its pods are slow to dehisce. Domestication has been associated with changes in the structure of pod valves and seed coats which reduce pod dehiscence and seed hardness. Pods and seeds have increased in size, mainly by increases in the rate of dry weight accumulation, and their increase has been only partly paralleled by increase in the area of subtending leaves. There has been no increase in the maximum photosynthetic rate of leaves, but the duration of their photosynthetic activity has increased. Domesticates are less sensitive than are wild plants to some environmental controls, such as in the response of germination to temperature, but in their flowering responses to daylength both wild and cultivated forms retain sensitivity under conditions where this is of adaptive value.     

Induction of 2n pollen in tulips by arresting the meiotic process with nitrous oxide gas

Triploid tulips have agronomically desirable traits such as vigorous growth and large flower size, but only a portion of all cultivated tulips is triploid. To apply 2n pollen to polyploid breeding of tulips, the polyploidizing agent, nitrous oxide gas (N₂O), was applied to bulbs. In tulips, meiosis in anthers occurs inside the bulbs from mid- to late-October. When meiosis in anthers (excised from bulbs) reached metaphase I, we treated other bulbs of the same clones with N₂O for 24–48 h. Most of the treated plants produced pollen grains with a wide-ranging or bimodal size distribution, indicating a mixture of n, 2n and aneuploid pollen grains. The use of pollen containing a relatively high proportion of giant pollen grains tended to yield larger numbers of triploids in the progeny. The number of giant pollen grains could be increased when N₂O-treated pollen grains were suspended in 10% sucrose and then sieved through a nylon mesh. Very few polyploids were observed in some cross combinations, even those involving pollen with a relatively high proportion of giant grains. Even so, this low polyploid yield most likely is due to a triploid block, because the capsules obtained in the crosses of the diploid×N₂O-treated plants contained some abnormal seeds, which were mostly triploid. Embryo culture was useful in rescuing abnormal embryos. The present study reveals that 2n pollen can be produced at high frequency using N₂O during tulip breeding.     

Tolerance to low temperatures and tuber soft rot in hybrids between Solanum commersonii and Solanum tuberosum obtained through manipulation of ploidy and endosperm balance number (EBN)

The objectives of this study were to evaluate the tolerance to low temperatures and tuber soft rot in hybrids between Solanum commersonii and Solanum tuberosum. The experimental materials consisted of F₁ triploid, BC₁ pentaploid‐near pentaploid and BC₂ tetraploid–near tetraploid hybrids. The F₁ triploids had a freezing tolerance and acclimatization capacity closest to S. commersonii. This indicated that the endosperm barriers which prevent the introgression of 1EBN S. commersonii into 4EBN S. tuberosum had been overcome. Indeed, the triploids produced 2n eggs, thus giving a compatible maternal to paternal EBN ratio in the hybrid endosperm generated by the 3x(2EBN) × 4x(3EBN) crosses. The tolerance to low temperatures of BC₁ and BC₂ hybrids was lower than that of the F₁. However, a number of genotypes were identified which were able to withstand temperatures down to ‐5°C. Some BC₂ hybrids were also tested for their tolerance to tuber soft rot, and some resistant hybrids were detected. A number of them combined the capacity for cold acclimatization with tolerance to tuber soft rot. These hybrids have an EBN of 4; they are fertile and have been used in backcrosses with 4EBN S. tuberosum.     

Intravarietal polyploidy in the apple (Malus pumila Mill.) cultivar Hazratbali

Summary A triploid cytotype of cv. Hazratbali (Malus pumila Mill.), was found growing in orchards of diploid trees. Morphological comparisons between the two cytotypes reveal that the triploid is more vigorous, and bears more attractive fruit. The meiotic behaviour of the two cytotypes has been described. The impact of polyploidy on quality characteristics of apple at the intravarietal level is analysed. The possible mode of origin of the triploid cytotype is discussed.     

Results of multiple pollination in blueberry (Vaccinium elliottii Chapm.)

Blueberry (Vaccinium section Cyanococcus, family Ericaceae) is a shrub that produces multiple-seeded berries in which only a fraction of the ovules develop into viable seeds. A recessive yellowleaf marker gene was used to evaluate the efectiveness of a single pollination versus multiple pollinations in producing seeds in Vaccinium elliottii Chapm. A multiple-pollination experiment was conducted to see if the first pollen applied or the second produced more progeny, and to see if multiple pollinations increased the number of seedlings produced. Flowers of diploid yellowleaf Vaccinium elliottii were pollinated from one to four times at daily intervals. Pollen from redleaf and yellowleaf plants was used to produce two types of seedlings that could be distinguished visually. Generally, percent berry set, seeds per berry, and seed germination were not increased by multiple pollination. After multiple pollination, the pollen applied first always produced the most seedlings. Prior pollination greatly reduced seed set from subsequent pollination, but subsequent pollination caused little or no decrease in the number of seeds set by the first pollination. This revised version was published online in July 2006 with corrections to the Cover Date.     

Barriers to hybridization of Solanum bulbocastanum Dun. and S. Verrucosum Schlechtd. and structural hybridity in their F1 plants

Summary Results of reciprocal crosses between Solanum verrucosum (2n=2x=24) and S. bulbocastanum (2n=2x=24) are described in terms of pollen tube behaviour in styles, of berry and seed set, of fertility and of meiotic behaviour of the F₁ hybrids. Pollen tube growth of S. verrucosum is strongly inhibited in styles of S. bulbocastanum, whereas no inhibition is observed in the reciprocal cross. Therefore S. bulbocastanum x S. verrucosum fails to set berries or seeds, whereas the reciprocal cross produces both berries (54.4% berry set) and seeds (0.3 per berry). Only 14.6% of the seeds germinate. Both the diploid and corresponding tetraploid hybrid plants are vigorous, flower abundantly, have a rather regular meiosis (mainly rod bivalents), but show a high degree of cytoplasmic-genic male sterility. Crossability of the diploid hybrid plants is nil when used as pollen parents and near to zero when used as pistillate parents. The barriers to hybridization of the parent species (unilateral inhibition of pollen tube growth, somatoplastic sterility, cytoplasmic-genic male sterility and structural differences of the parental chromosomes) are discussed and methods are suggested to overcome these barriers.     

A synthetic hexaploid (2n = 42) oat from the cross of Avena strigosa (2n = 14) and domesticated A. magna (2n = 28)

A synthetic hexaploid oat was produced by chromosome doubling of a sterile triploid hybrid between cultivated Avena strigosa (2n = 14) cv. Saia and a domesticated form of A. magna (2n = 28). The synthetic hexaploid was intermediate between its parents in panicle shape and lemma color, similar to the tetraploid parent in spikelet structure, and to the diploid parent in having a single, albeit partially shriveled seed per spikelet, and low protein content. By the third generation, plants with yellowish lemmas, mostly two seeds per spikelet and better filled grains had been selected. Rust resistance of the diploid parent was retained in the synthetic hexaploid, but not tolerance to barley yellow dwarf virus disease (BYDV). Chromosome associations at meiosis in the triploid hybrid was low, with over 60% of them being univalents. Bivalent association was the rule in the synthetic hexaploid with an occasional one or two quadrivalents. Regular meiosis with 21 bivalents was observed in further generations. The preferential pairing of homologous chromosomes in the synthetic hexaploid was probably contributed by the A. strigosa genome which exhibits this tendency in artificial allopolyploid situations. Selection of yellow lemma color and two seeds per spikelet suggests that the genes controlling these traits are located on the chromosomes involved in quadrivalents in the synthetic hexaploid. The potential and limitations of utilizing the synthetic hexaploid in oat research and breeding are briefly discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

The production of aneudihaploids in Solanum tuberosum L. group Tuberosum (the common potato)

Summary Trisomic cytotypes were produced in dihaploid (diploid) plants of Solanum tuberosum L. Group Tuberosum, the common potato, according to two methods. Firstly, the aberrant types were selected, through chromosome counting, from parthenogenic dihaploid offspring of tetraploid-diploid crosses. In dihaploid populations from twelve tetraploid potato varieties the frequencies of aneuploids ranged from 3.5 to 11.0%. About 95% of these aneuploids had only one, and the others not more than two extra chromosomes. Secondly, the aneuploids were produced from triploid-diploid crosses. Seedset strongly depended on the crossability of the parental plant material, and the best results were obtained when the motherplants were grafted onto tomato. On avarage the three most successful cross combinations resulted in approximately 0.7 berries per pollination and 6 seeds per berry. With regard to seedsize the seed could be divided in two groups, viz, normal and small seeds. Half of the seed did not germinate or produced inviable seedlings, especially among the small seed group. About 93% of the plants was neuploid, with 25, 26 and 27 being the predominant chromosome number. It was concluded that the production of trisomics was the most successful through triploid-diploid crosses. The results were discussed with reference to the chromosomal behaviour in the meiosis in triploid plants (Lange & Wagenvoort, 1973a). It thus was possible to relate the low seedset to the distribution of chromosomes in the meiotic anaphases. On the one side this distribution resulted in a limitation of the avalability of gametes with monohaploid and near-monohaploid chromosome numbers, while on the other side the macrospores with higher chromosome numbers seemed to be inviable. Finally a crossing scheme was presented for transmitting the trisomic condition into a genetic background with better homogeneity and more homozygosity.Extended version of a lecture presented at a Eucapia meeting about Potato Dihaploids, held at Wageningen (Netherlands). See Lange and Wagenvoort, 1973b.     

Successful induction of trigenomic hexaploid <Emphasis Type="Italic">Brassica</Emphasis> from a triploid hybrid of <Emphasis Type="Italic">B.</Emphasis><Emphasis Type="Italic">napus</Emphasis> L. and <Emphasis Type="Italic">B. nigra</Emphasis> (L.) Koch

A triploid hybrid with an ABC genome constitution, produced from an interspecific cross between Brassica napus (AACC genome) and B. nigra (BB genome), was used as source material for chromosome doubling. Two approaches were undertaken for the production of hexaploids: firstly, by self-pollination and open-pollination of the triploid hybrid; and secondly, by application of colchicine to axillary meristems of triploid plants. Sixteen seeds were harvested from triploid plants and two seedlings were confirmed to be hexaploids with 54 chromosomes. Pollen viability increased from 13% in triploids to a maximum of 49% in hexaploids. Petal length increased from 1.3 cm (triploid) to 1.9 cm and 1.8 cm in the two hexaploids and longest stamen length increased from 0.9 cm (triploid) to 1.1 cm in the hexaploids. Pollen grains were longer in hexaploids (43.7 and 46.3 μm) compared to the triploid (25.4 μm). A few aneuploid offsprings were also observed, with chromosome number ranging from 34 to 48. This study shows that trigenomic hexaploids can be produced in Brassica through interspecific hybridisation of B. napus and B. nigra followed by colchicine treatment.     

Crossability relationships of andean potato varieties of three ploidy levels

Summary The formation of triploids in 4x×2x crosses was studied in primitive potato varieties from the Andes of Peru and Bolivia. Triploid frequencies were as high as 80–100% in some crosses, although the actual number of seeds was low. The triploid block varied according to the tetraploid parent, and was never entirely absent. The likelihood of triploid formation was studied in 4x×2x crosses where pollen from both tetraploids and diploids of known genotype was used. Even though the diploids were chosen for their non-production of 2n gametes, as detected in pollen grain size, 66% of the hybrids were tetraploid. The amount and probability of gene-flow between the tetraploid and diploid gene-pools, either directly through 2n gametes or through triploid bridges was examined. Most of the 20 cultivars of S. x chaucha produced seed in crosses with tetraploids and diploids, although the fertility of the latter crosses was four times higher. The female fertility of some of the triploids thus allows them to exchange genes with the tetraploids and diploids, although gene-flow occurs more rapidly through direct transfer from diploids to tetraploids via 2n gametes.     

Investigation of the fenugreek (Trigonella foenum-graecum L.) genotypes for fresh weight, seed productivity, symbiotic activity, callus formation and aceumulation of steroids

Summary As a result of the analysis of 31 fenugreek (Trigonella foenum-graecum L.) genotypes we demonstrated that subsp. indica surpass significantly subsp. foenum-graecum in fresh weight, seed yield, nodulation ability, resistance to coot rot and the duration of the interstage and growing periods. There was no difference between the subspecies for leafiness and biochemical composition of seeds (concentrations of diosgenin and steroid glucosides, nitrogen, fat, inhibitors of tripsin and hemotripsin). The most promising genotypes (as compared with the standard cv. Nakhichevanskaya Shambala) yield 1650–1938 g/m² of fresh weight, 98.2–116.1 g/m² of seeds and accumulate 1.45–1.64% of diosgenin in seeds. High positive correlations (r=+0.53 ... +0.84; P_o<0.01) were found between fresh weight, seed yield, plant leafiness and height. Inoculation of the cv. Nakhichevanskaya Shambala with Rhizobium meliloti strains 851 and 852 increased its fresh weight by 94.5% and 93.6% and seed yield by 83.1% and 60.5%, respectively. Significant correlations of the nodule number with the fresh weight and seed yield (r=+0.70 and +0.77; P_o<0.01) allow us to assume that selection for increased symbiotic activity may be useful for improving the fenugreek productivity. The intersubspecies hybrids (four genotypes) surpass the subsp. indica (six genotypes) for the sizes of calli formed in presence of 2.4-D and kinetin. The sizes of calli formed by ten analysed genotypes in presence of BAP and kinetin (or 2.4-D) were negatively correlated with the nodule number (or fresh weight).     

Cross compatibility between diploid and tetraploid perennial ryegrass (Lolium perenne L.)

Summary A number of crosses were made between diploid and tetraploid Lolium perenne plants in order to determine the degree of cross compatibility between the two different ploidy levels. The range of hybridization undertaken involved diploidxdiploid, and tetraploid x tetraploid control crossings, diploid x tetraploid crosses, tetraploid x diploid crosses and mixed pollinations. The seed set, the germination capacity of resultant hybrid seeds, and the chromosome constitution of derived seedlings was determined. In addition attempts were made to culture 12 day-old hybrid embryos from the diploid x tetraploid and reciprocal crosses.The crossing results obtained indicated no barrier to fertilization between diploids and tetraploids, but the irregular and poor development of the seed accompanied by much reduced germination, indicated unsatisfactory endosperm establishment of large numbers of triploids, as confirmed by the result of culturing embryos dissected from 12-day old seeds.In preliminary studies undertaken to determine the extent to which diploid and tetraploid ryegrass varieties intercross when grown in close proximity for seed production, Griffiths and Pegler (1966) observed a very low incidence of triploids amongst the progenies of diploid plants exposed to the pollen of tetraploid plants and also amongst the progenies of tetraploid material exposed to the pollen of diploid plants.In these and subsequent studies it was noted that only a very small proportion of the so-called fully formed seeds derived from diploid x tetraploid crosses did actually germinate. When, in 1964, propagules of clones of S.24 perennial ryegrass, well separated from one another, were placed in drills of the tetraploid perennial ryegrass variety Reveille, the proportion of florets which formed caryopses, and of caryopses which germinated was as follows (Table 1).The data (Table 1) clearly indicate that a considerable proportion of the S.24 florets had been fertilized by 2n pollen and had formed caryopses, but most of these had proved to be defective. As noted in the previous studies, the frequency of triploid seedlings was low, thus suggesting incompatibility between the n and 2n gametes for the production of viable zygotes.Of the total progenies classed as tetraploids in the early seedling stage, 80% proved to be ryegrass x tall fescue F₁ hybrids. These arose as a result of free crossing with     

Structural hybridity in the series Etuberosa of the genus Solanum and its bearing on crossability

Summary Three non-tuberous Solanum species of series Etuberosa-S. brevidens (brd), S. etuberosum (etb) and S. fernandezianum (frn)-upon selfing displayed compatible berry set but seed set varied and on an average was below normal. Also in crosses between species berry set was unhampered (apart from the crosses with short-styled frn as pollen plant), but the interspecific hybrid plants showed various degrees of sterility and a low average seed set upon selfing. Cytological investigations revealed that in pure species-plants semi-sterility regularly occurred and was invariably associated with multivalent associations (up to decavalents). This may account for the decreased seed set found after selfing and intercrossing. Meiosis in F₁-hybrids was even more irregular, 75% of the pmc's showing multivalents involving from 4 to as many as 22 chromosomes in various complicated configurations in which even numbers of chromosomes were predominant. However, anaphase I separation was numerically balanced in more than 75% of the cells. Pollen stainability was very low in the hybrids.It is concluded that heterozygous translocations occur within each species and that the genomes of the species are differentiated by means of gross structural changes. In view of this and of the high chromosome pairing affinity the genome symbols E ₁, E ₂ and E ₃ are proposed for etb, brd and frn, respectively.Crossability of the Etuberosa species with tuberous S. pinnatisectum (pnt) was slightly more successful (average 2.7 seeds/berry) than that of Etuberosa species hybrids with S. pinnatisectum (average 1.7 seed/berry). In the hybrids studied-etb × pnt, frn × pnt and (brd × etb) × pnt-a high frequency of univalents was the rule (range 6–16 per cell). In addition rod bivalents (4–8 per cell) and a low frequency of trivalents occurred (0–3 in the biparental hybrids and 1–4 in the trispecific hybrid).Utilization of the valuable characters of Etuberosa for potato breeding is dependent on the degree of gene exchange between Etuberosa chromosomes and chromosomes of tuberous species. In the diploid hybrids such exchange, though limited, may take place through bivalents and trivalents. As shown in this article such diploid hybrids are non-crossable because of complete male and female sterility. Only after doubling the number of chromosomes the hybrids are fertile and can be used for further crosses.