An allotriploid derived from a amphidiploid × diploid

A fully fertile interspecific hybrid (Cucumis hytivus Chen and Kirkbride, 2n =4x =38) between Cucumis hystrix Chakr. (2n= 2x =24) and C. sativus L. (2n = 2x = 14) was previously produced by means of F₁ (2n = 19) embryo rescue and subsequent chromosome doubling. This amphidiploid, a new synthetic species, may serveas a genetic bridge in Cucumis, and thus is a source for broadening the genetic base of C. sativus. The identification and characterization of fertile progeny possessing lower ploidy levels would facilitate bridging among Cucumis species. Putative allotriploids (2n = 26) were recovered from C. hytivus × C. sativus matings by means of embryo culture, and experiments were designed to confirm their genetic constitution, describe their morphology, and establish an efficient protocol for their micropropagation. Apical and axillary buds of these putative allotriploid plants were used as explants to establish a micropropagation system for subsequent verification and characterization of ploidy. Of the array of micropropagation media tested, then ability to be most effective for the induction of adventitious buds (desginated Stage II) was a Murashige and Skoog (MS)growth media containing 13.3μM BA + 1.1μM NAA or containing10 μm BA only. The mean number of adventitious buds per explant in the two media was 6.8 and 6.5, respectively. Shoots resulting from adventitious buds produced roots (Stage III) in relative abundance (39 of 42, 92.8%) on half-strength MS medium containing 1.0 μm IBA. The survivorship of rooted plantlets after acclimatization as assessed by relative production of leaves in plantlets (designated Stage IV) was 91.4% (148 of 162). The chromosome number in putative allotriploid plants as determined in mitotic root tip figures in all plants was 2n = 26, the number expected for allotriploids derived from such a mating. An examination of pollen viability in five samples of each plant by cytochemical staining revealed stainability to be < %.Compared to their parents, the allotriploid genotypes possess a high degree of parthenocarpy (84.8%) as measured by setting fruit in pollen-free conditions. While allotriploid fruit are black-spined and similar to the maternal parent C. hytivus, the dark green leaves typical of allotriploid plants mirrors that of the paternal C. sativus parent. This revised version was published online in July 2006 with corrections to the Cover Date.     

Assessment of genetic relationships among Cucumis spp. by SSR and RAPD marker analysis

The first successful production of a sterile interspecific hybrid obtained from a cross between Cucumis hystrix Chakr. (2n = 2x = 24) and Cucumis sativus var. sativus L. (2n =2x = 14), and its subsequent fertility restoration through chromosome doubling provide an effective means for investigating genetic relationships among Cucumis spp. In this study, random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers were used to investigate relationships among C. s. var. sativus L., C. s. var. hardwickii (R.) Alef., C hystrix, C. hytivus Chen & Kirkbride (the amphidiploid species from chromosome doubling of the C. sativus x C. hystrix interspecific hybrid, 2n = 38), C. melo (2n =2x = 24) and C. metuliferus Meyer and Naudin (2n =2x= 24). A total of 109 SSR bands and 398 RAPD primed sites were used to calculate Jaccard's distance coefficients for cluster analysis using a unweighted pair‐group method using an arithmetic averaging (UPGMA) algorithm. The genetic relationships identified using SSR and RAPD markers were highly concordant, such that the correlation between SSR and RAPD genetic distance (GD) estimates was r = 0.94. SSR and RAPD analysis of 22 accessions allowed for their grouping into two distinct groups designated as CS and CM. While group CS consisted of 11 C. sativus genotypes, and the C. hytivus and C. hystrix accessions, group CM included six C. melo genotypes and C. metuliferus. The GD values between C. hystrix and C. sativus ascribed by SSR and RAPD matrices were 0.59 and 0.57, respectively. These GDs were smaller than those detected between C. hystrix and C melo (0.87 and 0.70 derived from SSR and RAPD markers, respectively).     

Synthesis and preliminary characterization of a new species (amphidiploid) in Cucumis

A successful interspecific hybridization between cucumber (Cucumis sativus L., 2n = 14) and a wild it Cucumis species, C. hystrix Chakr. (2n = 24) was made via embryo rescue. Hybrid plants (2n = 19; 7 from cucumber and 12 from C. hystrix) were sterile, but morphologically uniform. Self-pollination and backcrossing of F₁ hybrid plants to either parent confirmed presence of both male- and female-sterility that were likely caused by lack of homology and improper pairing during meiosis. While the multiple-branching habit, densely brown hairs (on corolla and pistil), orange-yellow corolla, and ovate fruit of F₁ hybrid plants were similar to that of the C. hystrix parent, the appearance of the first pistillate flower was more similar to that of C. sativus parent. The diameter and internode length of the stem, and the shape and size of leaves and flowers were intermediate when compared to the parents. The chromosome number in the hybrid was doubled through somaclonal variation during embryo culture and regeneration process to restore the fertility. Pollen grains were released and fruits with viable seeds matured on fertile, synthetic amphidiploid plants. The results from flow cytometry indicated that, on average, 7.3% of the morphologically unique regenerants had the 4C DNA content of 2.35 pg relative to the 2C DNA content of the original F₁ hybrid at 1.17 pg and, therefore, were likely chromosome-doubled F₁ hybrids (2n = 38). Nutrition alanalysis indicated that the synthetic species had higher protein (0.78%)and mineral (0.35%) content compared to the normal pickling cucumber(0.62% and 0.27%, respectively), and could be considered a new Cucumis crop having a special place in the future agriculture. Preliminarily evaluation indicates that C. hystrix possesses a high level of root-knot nematode resistance, and that this resistance is partially expressed in the interspecific F₁ and chromosome-doubled F₁. This and the fact that the fruit morphology of the fertile amphidiploid differs during the growing season (e.g., short and long fruit) suggest that it could be useful in broadening the germplasm base of cucumber. This revised version was published online in July 2006 with corrections to the Cover Date.     

Meiosis and fertility in interspecific hybrids of Nicotiana tabacum L. ×N. glauca Grah. and their derivatives

Nicotiana glauca is of potential interest to breeders as it carries resistance to black root rot of tobacco. Cytological investigations of sexual interspecific hybrids of N. tabacum T′T′TT (2n = 4x = 48) cv. ‘Wiślica’ × N. glauca GG (2n = 2x = 24) were carried out. The analyses of chromosome association at diakinesis and metaphase I in the PMCs of amphihaploid F₁ T′TG (2n = 3x = 36) revealed low variable pairing with 0–5 bivalents. The sterile amphihaploids F₁ were converted into partial female fertile amphidiploids T′T′TTGG by chromosome doubling. Among 36 mature plants obtained, 15 were found to have chromosome numbers (2n = 6x = 72) and were verified as amphidiploids, 9 had (2n = 6x = 70 or 71) chromosomes while the remaining 12 were haploid. True amphidiploids, in spite of quite high chromosome pairing during meiosis, were very different in pollen fertility, ranging from 0% to 85%. Male fertility disturbances did not correlate with the degree of female fertility upon pollination with N. tabacum. Sesquidiploids T′TG (2n = 5x = 60) obtained from backcrossing the amphidiploids to parental tobacco showed more than 22 bivalents, 10–12 univalents and occasional multivalents that indicated the possibility of interchange events between N. tabacum and N. glauca genomes.     

Cytogenetics of a new trispecies hybrid in cotton: [(Gossypium hirsutum L. × G. thurberi Tod.)2 × G. longicalyx Hutch. & Lee]

A three‐species hybrid named HTL including Gossypium hirsutum L. [2n = 4 x = 52, (AD)₁ genome] was created using the pseudophyletic introgression method with G. longicalyx Hutch. & Lee (2n = 2x = 26, F₁ genome) as donor parent and G. thurberi Tod. (2n = 2x = 26, D₁ genome) as bridge species. The new hybrid was totally self‐sterile and its interspecific status was confirmed using simple sequence repeat markers and cytogenetic analysis. Cytogenetic studies showed that its chromosome configuration was 2n = 52 = 14.13 I + 15.10 II + 1.03 III + 0.9 IV + 0.03 V + 0.13 VI (where I, II, III, IV, V and VI are univalents, bivalents, trivalents, tetravalents, pentavalents and hexavalents, respectively). Prospects for successfully exploiting the HTL hybrid in breeding programmes are discussed.     

Incompatibility in diploid and tetraploid crosses of Cucumis sativus and Cucumis metuliferus

The African horned cucumber (Cucumis metuliferus Naud.; 2x = 2n = 24) contains genes that can confer resistance to many important cucumber (C. sativus L.; 2x = 2n = 14) pests [e.g., root-knotnematode, Meloidogyne incognita (Kofoid & White) Chitwood]. Cucumber is highly susceptible to this root-knot nematode species, and a recent screening of C. sativus accessions in the U.S. National Plant Germplasm collection did not identify sources of resistance. Thus,autotetraploids of Cucumis sativus and C. metuliferus were created to recover fertile resistant interspecific progeny. Autotetraploids were obtained at the highest rate when seeds were immersed in 0.5% colchicine for a period of 6 to 8 hrs. Treatment durations less than 6 hrs produced few tetraploids, and durations of 10 hrs or more were lethal to seeds or developing seedlings. Crosses between C. sativus and C. metuliferus were made using diploid and tetraploid lines in all possible combinations, including reciprocals. Fruit development occurred in crosses when diploid and tetraploid C. sativus were used as the female parent. However, seeds developed only in fruit of C. sativus (4n) ×C. metuliferus (2n) crossings. Seeds from these crosses, however,were flat and not viable. No fruit development occurred in crosses whereC. metuliferus was used as the female parent. This revised version was published online in August 2006 with corrections to the Cover Date.     

Occurrence and chromosome number of Cucumis sativus var. hardwickii (Royle) Alef. in South India and its bearing on the origin of cultivated cucumber

Summary Cucumis sativus var. hardwickii (Royle) Alef., the wild progenitor of cultivated C. sativus is reported for the first time from peninsular India. The South Indian specimens showed n=7 bivalents in PMCs. The discontinous occurrence of the wild taxon in the Himalayan regions and peninsular hills and the existence of cultivars of C. sativus adapted to the tropical and temperate climates suggest polytopic domestication of the cultivated forms. The possibility of utilizing this wild germplasm for crop improvement is indicated.     

Unique meiotic behaviourin F1 plants from a cross between a non-tuberous and a tuberous Solanum species in section Petota

Uniformly abnormal meiotic behaviour was observed in 12 F₁-plants from a cross between Solanum etuberosum (non-tuberous) and S. pinnatisectum (tuberous). Per pollen mother cell at MI an average was found of 3.64 bivalents (all rodshaped, 1–2 per cell, heteromorphic), 16.64 univalents (scattered haphazardly on a continuous bipolar spindle) and 0.03 trivalents (all Y-shaped). Lagging chromosomes and precocious division of univalents very frequently occurs, leading to unequal distribution of chromosomes, aneuploid gametes and male sterility. Heteromorphic bivalents at MI, loops in bivalents at pachytene and non-disjunction in one hybrid plant, point to a highly abnormal meiotic behaviour. The occurrence of few trivalents is discussed.Considering that according to the literature nearly normal pairing was observed in the intergeneric F₁-hybrids Lycopersicon esculentum × Solanum lycopersicoides and L. esculentum x S. pennellii, the lack of chromosome pairing in an interspecific F₁-hybrid, of which both Solanum parents belong to the same section, is paradoxical to a plant breeder and might even be conspicuous to a taxonomist.     

Cyto-morphological evidence for segmental allopolyploid origin of Teasle gourd (<Emphasis Type="Italic">Momordica subangulata</Emphasis> subsp. <Emphasis Type="Italic">renigera</Emphasis>)

Teasle gourd [Momordica subangulata Blume subsp. renigera (G. Don) de Wilde, 2n = 56] exhibits morphological characters found in both M. dioica (2n = 28) and M. cochinchinensis (2n = 28). Morphological analysis of M. subangulata subsp. renigera suggests an allopolyploid origin. We present evidence elucidating the genomic relationships between M. dioica, M. cochinchinensis and M. subangulata subsp. renigera. A triploid M. dioica × M. subangulata subsp. renigera hybrid had an average of 12.76 bivalents, 13.84 univalents and 0.88 trivalents at metaphase I, while the M. cochinchinensis × M. subangulata subsp. renigera hybrid had an average of 13.08 bivalents, 12.96 univalents and 0.96 trivalents. F₁ hybrids of the two diploid species (M. dioica × M. cochinchinensis) showed an average of 9.12 bivalents and 9.76 univalents, suggesting that the genomes of these species are only partially homologous. A higher number of bivalents in the triploid hybrids suggests that M. subangulata subsp. renigera is a segmental allopolyploid of M. dioica and M. cochinchinensis and that its genomes have diverged from the parental genomes.     

Synthesis and sterility of raphanobrassica

Summary The synthesis of Raphanobrassica (2n=36, rrcc) from Raphanus sativus (2n=18, rr) and Brassica oleracea (2n=18, cc) is described a) by colchicine treatment of diploid hybrids; b) by crossing autotetraploid froms of the parent species.The variation within R. sativus and B. oleracea suggests that a range of morphologically distinct Raphanobrassica forms may be created, some of which may have agronomic potential and in particular, it is hoped, Plasmodiophora resistance.Inter-generic hybrids were readily obtained from crossing the parental species at both 2x and 4x chromosome levels, but only with R. sativus as female parent.Details are given of the morphology, fertility and chromosome behaviour of both diploid F₁ R. sativus × B. oleracea hybrids and of the amphidiploid Raphanobrassica.Synthesized Raphanobrassica plants proved, in general, highly sterile. Some aneuploids resulted from 4x R. sativus × 4x B. oleracea crosses but most progeny were euploid and showed almost regular chromosome association. A number of stunted, deformed plants were obtained from both 2x and 4x crosses. Vigour, fertility and aneuploidy appeared unconnected in the amphidiploid.Previous work on Raphanobrassica is reviewed. It is concluded that the extremely low fertility encountered in the present study is more likely to be the result of genic imbalance than to cytological anomalies which appear to be of lesser significance.     

Monohaploids (n=x=12) from autotetraploid Solanum tuberosum (2n=4x=48) through two successive cycles of female parthenogenesis

Summary Viable monohaploids with 12 chromosomes in their somatic cells were obtained from an autotetraploid cultivar of Solanum tuberosum (2n=4x=48) by inducing female parthenogenesis successively in the tetraploid cultivar and in dihaploids from that cultivar. Both dihaploids and monohaploids were induced using the diploid S. phureja clones IvP 35 and IvP 48 as pollinators. The average frequencies of dihaploids and monohaploids in 1973 were 2.0 and 0.14 per berry respectively.Non-homologous chromosome associations (bivalents, trivalents and even quadrivalents) were observed at metaphase I of meiosis in pollen mother cells of the two monohaploids studied.The occurrence of non-homologous associations of chromosomes during meiosis is discussed. In addition the potential significance of monohaploids for basic research andfor breeding of potatoes is considered.     

Production and characterization of Raphanus sativus-Brassica rapa monosomic chromosome addition lines

Breeding of Raphanus sativus‐Brassica rapa monosomic chromosome addition lines (MALs, 2n = 19) was carried out by backcrossing the synthesized amphidiploid line, Raphanobrassica (R. sativus×B. rapa, 2n = 38, RRAA, line RA89) with R. sativus cv. ‘Shogoin’ (2n = 18, RR). In the first cross of Raphanobrassica× radish, four sesquidiploidal BC₁ plants (2n = 28, RRA, RA89‐36‐1, RA89‐31‐1, RA89‐31‐2, RA89‐31‐3) were successfully developed. In these plants, the chromosome configurations of 9II + 10I and 10II + 8I were observed frequently at first metaphase (MI) of meiosis in pollen mother cells (PMCs). The RA 89‐36‐1 plant produced many seeds in the reciprocal backcrosses with radish. About 50% of the BC₂ plants obtained from the cross of RA89‐36‐1 plant × radish were 2n = 19 plants, followed by 2n = 18 plants (24%) and 2n = 20 plants (19%). In the reciprocal cross, 2n = 19 plants were also developed at the rate of 40%. From analysis of specific morphological traits, 2n = 19 plants were classified into eight types (a‐h). When 25 selected primers were used in polyacrylamide gel electrophoresis, random amplified polymorphic DNA (RAPD) markers derived from B. rapa for each type of MAL were detected in numbers between three for e‐type and 16 for b‐type. RAPD markers specific for each type alone were from one (OPE 05‐344) for h‐type to nine for b‐type. In the g‐type, no marker specific to this type alone was observed. However, 19 bands were common between at least two types. These MAL plants exhibited predominantly the chromosome configuration of 9II + 1I at MI of PMCs, pollen and seed fertility being the same level as the radish cv. ‘Shogoin’. From the morphological traits and DNA markers, eight different MAL types among 10 expected were identified.     

Marker-assisted backcross selection in an interspecific <Emphasis Type="Italic">Cucumis</Emphasis> population broadens the genetic base of cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis> L.)

Cucumber (Cucumis sativus L.) is a major cucurbit vegetable species whose genetic base has been drastically reduced during its domestication. The crop’s narrow genetic base (3–12% DNA polymorphism) has resulted from the use of limited genetic material and intense selection during plant improvement. Recently, however, interspecific hybridization has been successful in Cucumis via mating of C. hystrix Chakr. and C. sativus, which resulted in the amphidiploid C. hytivus. We report herein a marker-assisted strategy for increasing genetic diversity in cucumber through introgression backcrossing employing C. hytivus. The comparatively late-flowering but high-yielding, indeterminate, monoecious line WI 7012A (P₁; donor parent) derived from a C. hytivus × C. sativus-derived line (long-fruited Chinese C. sativus cv. Beijingjietou) was initially crossed to the determinate, gynoecious C. sativus line WI 7023A (P₂; recurrent parent 1), and then advanced backcross generation progeny (BC₂) were crossed with the gynoecious indeterminate line WI 9-6A (P₃; recurrent parent 2). More specifically, a single F₁ individual (P₁ × P₂) was backcrossed to P₂, and then BC progeny were crossed to P₂ and P₃, where marker-assisted selection (MAS) for genetic diversity (8 mapped and 16 unmapped markers; designated Sel) or no selection (designated NSel) was applied to produce BC₃P₂ (Sel) and BC₃P₃ (Sel), and BC₂P₂ (NSel) and BC₂P₂S₁ (NSel) progeny. Relative vegetative growth, number of lateral branches (LB), days to flowering (DF), yield (fruit number), and fruit quality [as measured by length:diameter (L:D) and endocarp:total diameter (E:T) ratios] were assessed in parents and cross-progeny. DF varied from ~20 (BC₃P₂Sel) to ~25 days (BC₂P₃Sel) among the populations examined, where progeny derived from P₂ possessed the shortest DF. Differences in cumulative yield among the populations over six harvests were detected, varying from ~8 fruits per plant in BC₃P₂ (Sel) to ~39 fruits per plant in BC₂P₃ (Sel). Although the vigorous vegetative growth of line P₁ was observed in its backcross progeny, highly heterozygous and polymorphic backcross progeny derived from P₃ were comparatively more vigorous and bore many high-quality fruit. Response to selection was detected for LB, DF, L:D, and E:T, but the effectiveness of MAS depended upon the parental lines used. Data indicate that the genetic diversity of commercial cucumber can be increased by introgression of the C. hystrix genome through backcrossing.     

Genomic Affinity Relationships in the Genus Avena

Many interspecific hybrids between diploid, tetraploid and hexaploid Avena species were obtained without artificial embryo cultures. Most of the hybrids showed variable chromosome pairing with univalents, bivalents and multiple associations, even in PMC in the same hybrid, at first metaphase; making ii very difficult to understand the degree of genome affinity between parent species, and especially between different species. The basic or theoretical chromosome pairing consisting of bivalents and univalents is presented, in which the chromosome ratios (%) statistically agree with observations. In addition, the per cent frequency of chromosomes forming multivalents is calculated. On the bases of these numerical data, the genomic affinity relationships are discussed for present and previously reported interspecific hybrids or Avena.     

Genetic stability and maintenance of Raphanus sativus lines with an added Brassica rapa chromosome

The genetic stability and maintenance of Raphanus sativus‐Brassica rapa monosomic chromosome addition lines (a‐h‐types MALs, 2n = 19, BC₂), developed by backcrossing the synthesized amphidiploid Raphanobrassica (Raphanus sativus × Brassica rapa, 2n = 38, RRAA) with R. sativus cv. ‘Shogoin’ (2n = 18, RR), was investigated. Transmission of the added alien chromosome through selected smaller seeds (SSS) and the inheritance of morphological traits and random amplified polymorphic DNA (RAPD)‐specific markers together with meiotic chromosome configuration and seed fertility were also investigated for three successive generations (BC₃ to BC₅). The distinctive traits and the RAPD‐specific markers of the eight types of MAL were substantially inherited and stably maintained throughout three generations, although a few variant plants (2n =18) resembling MALs (2n = 19) and hyperploidal plants (2n = 26 and 2n = 37) were generated in the earlier generations of BC₃ and BC₄ in comparison with BC₅. The average transmission rates for three generations ranged from 26% for both the b‐type and the d‐type to 44% for the e‐type through SSS. On the other hand, the transmission rates through randomly selected seeds (RSS) were lower, ranging from 6.5% for the f‐type to 23.5% for the b‐type. In meiosis, more than 90% of PMCs showed the 9II +1I pairing configuration at metaphase I throughout three generations. For seed fertility, when backcrossed with the radish cv. ‘Shogoin’, the values were approximately 180% to 500% with the mode around 300% with the seed harvested from a pod increasing with the advancing generations. Genetic recombination between the radish chromosomes and the added chromosome is probably rare, suggesting that the added chromosome is mostly maintained unaltered in the background of the radish genome.     

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.     

Analysis of stability for some characters in Kabuli Chickpea

Summary The meiotic behaviour of a hybrid between Triticum aestivum and the amphiploid Hordeum chilense x T. turgidum conv. durum, was studied using a C-banding staining method. This hybrid has the genome formula of AA BB D Hch with 2n=6x=42 chromosomes. The durum wheat chromosomes (genomes A and B) were easily recognized, whereas the D and Hch chromosomes were recognized as a whole. Meiotic pairing was homologous, as expected (14 bivalents from A and B genomes +14 univalents from D and Hch genomes). However, some pollen mother cells at metaphase-I presented pseudobivalents that could have been caused by either homoeologous or autosyndetic pairing amongst D and Hch chromosomes.     

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.     

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.     

New C-banding pattern for chromosome identification in cucumber (Cucumis sativus L.)

A chromosome study of cucumber, C. sativus L., was performed using orcein and C-banding techniques. The diploid and tetraploid plants investigated here showed the somatic chromosome numbers 2n=14 and 28, respectively. The haploid chromosome complement was composed of five metacentric and two submetacentric chromosomes. All C. sativus chromosomes had clearly visible C-bands, and each chromosome could be identified unequivocally after C-banding staining, with 13 C-bands appearing in the haploid complement. The haploid complement had a 44.9% ratio of total C-band length to total chromosome length. Chromosomes 1, 2, 4, 5 and 7 had stable C-bands. Three large, dark C-bands appeared at the proximal regions of chromosomes 1 and 2. Chromosome I had quite a large C-band and with a 68.4% ratio of C-band length to short arm length. Chromosome 2 also had quite a large C-band in the pericentromeric region with a 57.6% ratio of C-band length to the full length of this chromosome and possessed an elongated primary constriction in early metaphase. In prometaphase, chromosome 2 showed that the long arm was completely separated from the short arm. The number of secondary constrictions could not be clearly observed because these chromosomes are small and they could not be counted in every metaphase cell. However, six chromosomes seemed to have secondary constrictions in the diploid plants. Two silver-stained bands were observed at primary constrictions of two of the large chromosomes.