Induction of an alloplasmic male-sterile Brassica oleracea by substituting cytoplasm from ‘Early Scarlet Globe’ radish (Raphanus sativus)

Summary Alloplasmic male-sterile Brassica oleracea L. was synthesized in a backcrossing program through amphidiploid Raphanobrassica by using Early Scarlet Globe radish (Raphanus sativus L.) as the donor of cytoplasm and B. oleracea broccoli and cabbage as recurrent pollen parents. Persistence of radish chromosomes and high female sterility were encountered in the first four backcrosses. Following use of colchiploid 4x broccoli as pollen parent, a BC₅ plant was obtained that had 2n=3x+1=28 chromosomes, improved seed set, and no radish traits. The BC₆ with recurrent 2x broccoli contained male-sterile plants with 2n=18 or 19 chromosomes, increased seed set, and broccoli morphology. Subsequent generations segregated for male-sterile and restored male-fertile plants, some with variable development of stamens and pollen. Leaf color of the alloplasmic plants, especially seedlings, was lighter green than normal.     

Chromosomal and genic structure of Brassicoraphanus related to seed fertility and the presentation of an instance of improvement of its fertility

Summary In order to elucidate the mechanism of low fertility of Brassicoraphanus, i.e., amphidiploids between Brassica japonica Sieb. and Raphanus sativus L., the chromosome number of 253 plants was studied during the 3rd–9th generations for their seed fertility. Meiotic irregularity showed no connection with degree of sterility. Brassicoraphanus consisted of euploids (2n=38), hyperploids (2n=39–43) and hypoploids (2n=34–37) with white or yellow flowers. The number of plants was highest in euploids and became lower as the chromosome number diverged from the euploid number. Further, seed fertility was highest and the range of its variation widest in euploids. The seed fertility of aneuploids became lower and its variation narrower in proportion to the number of chromosomes additional to or missing from the euploid number. Yellow-flowered plants were superior in seed fertility to white-flowered plants. Seed fertility of plants is primarily affected by their chromosome numbers and secondarily modified by genic effects. As a whole, seed fertility of Brassicoraphanus increased gradually and its variation widened with the advance of generations. This was explained mainly by the increase of balanced combinations of genes.     

The increase of seed fertility of Brassicoraphanus through cytological irregularity

Summary Amphidiploids (Brassicoraphanus) were produced by means of colchicine treatment of F₁ hybrids between Brassica japonica Sieb. and Raphanus sativus L. The cytology of the amphidiploids was studied from F₁ to F₃ generations. Some plants had the euploid chromosome number 2n=38, whereas others had the aneuploid number 2n=37. One or two of either quadrivalents or trivalents, as well as some univalents, were seen in most of the plants examined. All the plants showed a low seed fertility. In F₃ generation there arose some yellow-flowered plants, all of which showed a higher seed fertility than normal white-flowered plants. It is postulated that the change of flower colour might originate in the segmental exchange of only partially homologous chromosomes following multivalent formation. A gene causing white flower colour was perhaps closely linked to a gene causing sterility, and both genes were probably excluded together through the segmental exchange of the chromosomes. Therefore, it can be said that the increase of fertility was induced by cytological irregularity.     

Intergeneric crosses for the transfer of resistance to the beet cyst nematode from Raphanus sativus to Brassica napus

Summary The possibilities to transfer important traits and in particular resistance to the beet cyst nematode (Heterodera schachtii, abbrev. BCN) from Raphanus sativus to Brassica napus were investigated. For these studies B. napus, R. sativus, the bridging hybrid ×Brassicoraphanus (Raparadish) as well as offspring of the cross ×Brassicoraphanus (Raparadish) ×B. napus were used. Reciprocal crosses between B. napus and R. sativus were unsuccessful, also with the use of embryo rescue. Crosses between ×Brassicoraphanus as female parent and B. napus resulted in a large number of F₁ hybrids, whereas the reciprocal cross yielded mainly matromorphic plants. BC₁, BC₂ and BC₃ plants were obtained from backcrosses with B. napus, which was used as the male parent. F₁ hybrids and BC plants showed a large variation for morphology and male and female fertility. Cuttings of some F₁ and BC₁ plants, obtained from crosses involving resistant plants of ×Brassicoraphanus, were found to possess a level of resistance similar to that of the resistant parent. These results and indications for meiotic pairing between chromosomes of genome R with those of the genomes A and/or C suggest that introgression of the BCN-resistance of Raphanus into B. napus may be achieved.     

Interspecific hybrids between helianthus PetiolarisNutt. and H. Annuus L.: Effect of backcrossing on meiosis

Summary Interspecific substitutions of the nucleus of Helianthus annuus (2n=34) into the cytoplasm of H. petiolaris (2n=34) were obtained by successive backcrossing using cultivated sunflower, H. annuus, as the recurrent pollen parent.Meiosis in the F₁ was characterized by multivalents, suggesting that 10 of the 34 chromosomes were heterozygous for chromosomal interchanges. An additional pair of chromosomes also contained a paracentric inversion. Continued backcrossing resulted in rapid elimination of the meiotic aberrations evident in the F₁. In the BC₁, 1 of 11 plants had normal meiosis and by the BC₂, only 13 of 54 plants had meiotic aberrations similar to those of the F₁. However, trisomic progeny (2n=35) were found in 3 of the 11 BC₁ plants and 20 of the 54 BC₂ plants. No meiotic aberrations were observed in BC₃ or BC₄ plants. Plants with indehiscent anthers, and considered to be male sterile (M.S.), first occurred in the BC₁ and, by the BC₂, 51 of 54 plants were M.S. All 19 BC₃ and 16 BC₄ plants were M.S. Preliminary investigations suggest that the pollen from such plants is sterile and that the sterility is cytoplasmic rather than genetic.Disc-flower measurements were a useful technique for selecting samples at the correct stage of microsporogenesis, but could not be used to distinguish between successive backcrosses.     

The effect of introgressions of wheat D-genome chromosomes into 'Presto' triticale

Hexaploid triticale (X Triticosecale Wittmack) (2n= 6x= 42, AABBRR) and wheat (Triticum aestivum L.) (2n= 6x= 42, AABBDD) differ in their R and D-genomes. This produces differences in both agronomic and end-use quality characteristics. Our objective was to determine how introgressions of individual chromosomes from the D-genome of wheat affect these characteristics of a winter triticale 'Presto'. We studied the effects of 18 D-genome chromosome substitution lines, 15 sib-lines as controls, and five check cultivars at Lincoln, NE in 1996, using a randomized complete block design with two replications. The experiment was repeated at Lincoln and Mead, NE in 1997 and 1998 with 15 substitution lines that survived the first winter in Lincoln, along with their 12 control sibs and five check cultivars. Few D-genome chromosomes had positive effects. Chromosomes 2D, 4D, and 6D significantly reduced plant height when substituted for 2R, 4B, and 6R, respectively. No grain yield increases were associated with any of the D-genome chromosomes tested, but three substitutions decreased the grain yield. Depending on the allele of the hardness gene present, chromosome 5D increased or decreased kernel hardness when substituted for 5R or 5A, respectively. Introgressions of chromosomes 1D and 6D improved end-use quality characteristics of Presto. These results suggest that apart from beneficial effects of individual loci located on the D-genome chromosomes, no major benefit can be expected from D-genome chromosome substitutions.     

Production of intergeneric hybrids between Brassica juncea and Diplotaxis virgata through ovary culture, and the cytology and crossability of their progenies

The cytogenetic study was investigated in the intergeneric F₁ hybrid, F₂and backcross progenies (BC₁). The plants used were Brassica juncea(2n=36) and Diplotaxis virgata(2n=18). Three intergeneric F₁ hybrids between two species were produced through ovary culture. They showed 36 chromosomes. It might consist one genome of B. juncea and two genomes of D. virgata. The morphology of the leaves resembled that of B. juncea. The color of the petals was yellow that was like in D. virgata. The size of the petal was similar to that of B. juncea. The mean pollen fertility was15.3% and the chromosome associations in the first meiotic division were(0–1)_IV+(0–2)_III+(8–12)_II+(12–20)_I. Many F₂ and BC₁seeds were harvested after open pollination and backcross of the F₁ hybrids withB. juncea, respectively. The F₂seedlings showed different chromosome constitutions and the range was from 28 to54 chromosomes. Most seedlings had 38chromosomes followed by 36, 40 and 54. The BC₁ seedlings also showed different chromosome constitutions and the range was from 29 to 62. Most seedlings had both 40and 54 chromosomes followed by 36, 46 and52. In the first meiotic division of F₂ and BC₁ plants, a high frequency of bivalent associations was observed in all the various kinds of somatic chromosomes. Many F₃ and BC₂ seeds were obtained by self-pollination and open pollination of both F₂ and BC₁ plants, and by backcrossing both F₂ and BC₁plants with B. juncea, respectively,especially, three type progeny with 36, 40or 54 chromosomes. The somatic chromosomes of the F₃ and BC₂ plants were further investigated. The bridge plants between B. juncea and D. virgata with 36 chromosomes may be utilized for breeding of other Brassica crops as well as B. juncea. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

A new source of cytoplasmic male sterility in sunflower

Summary Interspecific substitutions of the nucleus of Helianthus annuus (2n=34) cv. Saturn into the cytoplasm of H. petiolaris (2n=34) by successive backcrossing, resulted in progenies with indehiscent anthers containing white, rather than normal yellow, pollen. Further backcrossing by cv. Saturn failed to restore pollen shed, suggesting that the male sterility was cytoplasmic. In vivo germination tests of pollen from 23 such plants from eight BC₅ lines, indicated complete pollen sterility for 14 plants, but normal seed set, suggesting that female fertility was not affected. Meiosis in all plants examined was normal.Crosses between seven sources of pollen-fertility restorer, one collection of wild H. annuus, and an existing source of cytoplasmic male sterility, resulted in a high frequency of plants with normal pollen shed in all F₁ progenies. However, no normal pollen shed was evident in F₁ progenies for similar crosses between BC₅ male-steriles and three of the seven restorer sources, nor for the single wild H. annuus evaluated. The foregoing suggests that the backcross substitution lines are a new source of cytoplasmic male sterility. The inheritance of restoration of pollen shed was complex and not fully elucidated. Some data suggested that two independent, complementary, dominant genes were required, but others indicated two to three independent, dominant genes.     

Glandular hair densities in three perennial Medicago species

Summary Eight triazine resistant (Brassica napus x B. oleracea) x B. oleracea interspecific hybrids with chromosome numbers ranging from 25 to 27 were backcrossed a second time to B. oleracea but no seed was formed. However, in vitro embryo rescue on 77 developing ovules yielded nine BC₂ plants with chromosome numbers between 19 and 25 and in which the herbicide resistance was still strongly expressed. Three of these plants (NOH-8B2B1, 2n=20; NOH-8B2B3 and NOH-8B2B4, 2n=19) were backcrossed again to B. oleracea. Two of the three plants produced seed which germinated to produce triazine resistan BBC₃s with 18, 19 or 20 chromosomes. The triazine resistant B. campestris cytoplasm has now been stabilized in B. oleracea.     

Interspecific cytoplasm substitutions of an Indica strain of Oryza sativa L. and O. Glaberrima Steud

Summary To study differential nucleus-cytoplasm interactions between the two cultivated rice species, Oryza sativa and O. glaberrima, cytoplasmic substitution lines were made by using a glaberrima strain (G) and an Indica strain of sativa (S). The G cytoplasm had no adverse effect on pollen development when combined with the nucleus of S. On the other hand, when the S cytoplasm was combined with the G nucleus, the substitution line showed no seed set because of male sterility although the pollen grains were normally stained with I₂-KI solution. A dominant gene derived from S strain seemed to cause anther indehiscence in the substitution line. Further, a restorer gene (Rf _j)from Akebono of Japonica type was effective on pollen restoration in the male sterile line, suggesting that the S cytoplasm is the same as those of Japonica type in terms of a fertility-restoring system.This paper is Genetic studies of speciation in cultivated rice. 4.     

Molecular mapping and detection of the yellow rust resistance gene Yr26 in wheat transferred from Triticum turgidum L. using microsatellite markers

Yellow rust (stripe rust), caused by Puccinia striiformis Westend f. sp. tritici, is one of the most devastating diseases of wheat throughout the world. Wheat-Haynaldia villosa 6AL.6VS translocation lines R43, R55, R64 and R77, derived from the cross of three species, carry resistance to both yellow rust and powdery mildew. An F₂ population was established by crossing R55 with the susceptible cultivar Yumai 18. The yellow rust resistance in R55 was controlled by a single dominant gene, which segregated independently of the powdery mildew resistance gene Pm21 located in the chromosome 6VS segment, indicating that the yellow rust resistance gene and Pm21 are unlikely to be carried by the same alien segment. This yellow rust resistance gene was considered to beYr26, originally thought to be also located in chromosome arm 6VS. Bulked Segregation Analysis and microsatellite primer screens of the population F₂ of Yumai 18 × R55 identified three chromosome 1B microsatellite locus markers, Xgwm11, Xgwm18 and Xgwm413, closely linked to Yr26. Yr26 was placed 1.9 cM distal of Xgwm11/Xgwml8, which in turn were 3.2 cM from Xgwm413. The respective LOD values were 21 and 36.5. Therefore, Yr26 was located in the short arm of chromosome 1B. The origin and distribution of Yr26 was investigated by pedigree, inheritance of resistance and molecular marker analysis. The results indicated that Yr26 came from Triticum turgidum L. Three other 6AL.6VS translocation lines, R43, R64 and R77, also carried Yr26. These PCR-based microsatellite markers were shown to be very effective for the detection of the Yr26 gene in segregating populations and therefore can be applied in wheat breeding. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

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.     

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.     

The mechanism of increased seed fertility accompanied with the change of flower colour in Brassicoraphanus

Summary In Brassicoraphanus (amphidiploids between Brassica japonica Sieb. and Raphanus sativus L.), yellow-flowered plants that occurred among originally white-flowered plants showed an increased seed fertility. It is assumed that the gene Y (yellow-flower gene) from Brassica and the gene W (white-flower gene) from Raphanus are located at corresponding loci of only partially homologous chromosomes. W is dominant (epistatic) over Y. The normal white-flowered plants have the genotype YYWW. A YYYW-plant was found, which is assumed to have arisen through crossing-over following multivalent formation. In the progeny of this plant, yellow-flowered plants (YYYY) as well as white-flowered plants (YYWW, YYYW) appeared. The gene for flower colour is closely linked to a gene which controls the development of embryos (or endosperm). This gene promotes the development of embryos in homozygous condition. Therefore, the embryo having only the yellow-flower gene can develop more easily into viable seed than the embryo having the white-flower gene. It is also possible that the sterility of white-flowered plants is caused by a discordance between the cytoplasm of Brassica and W (or genes linked to W) of Raphanus.     

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.     

Development of new cytoplasmic genetic male sterile lines through Indica × Japonica hybridization in rice

Summary From 28 Indica-Japonica crosses, two Indica cultivars, V.20B and Sattari were identified to possess male sterile cytoplasm with fertility restoring genes. It was possible to develop a new Japonica cytoplasmic genetic male sterile line (Zhunghua-1) on Indica male sterile cytoplasm (V 20B) by repeated backcrossing the complete pollen sterile plants of V 20B x Zhunghua-1 to the recurring male parent, Zhunghua-1. The study indicated that it would be possible to develop male sterile lines rom indica-japonica crosses only when there is sufficient amount of reciprocal differences with respect to pollen sterility. Further, it was inferred that it would be easier to develop Japonica male sterile lines on Indica cytoplasm than developing Indica male sterile line with japonica cytoplasm.     

Cytomorphological studies in an intergeneric hybrid between Gossypium hirsutum L. (2n=52) and Hibiscus panduraeformis Burm.

Summary An intergeneric hybrid (2n=38) between Gossypium hirsutum L. (2n=52) × Hibiscus panduraeformis Burm. (2n=24) was obtained by pollinating about 2000 flower buds of G. hirsutum var. Gregg Male Sterile with pollen from H. panduraeformis. The F₁ hybrid was intermediate in plant habit, but possessed gossypol glands and nectaries on the leaves, bolls containing seeds with fuzz and lint as dominant characters of G. hirsutum. Flowers with yellow corolla and anthers; purple petal spot, profuse growth of epidermal hairs on all plant parts including the boll sutures, and jassid tolerance were dominant characters of H. panduraeformis. The partial fertility of the F₁ indicated the possibilities of combining jassid and drought tolerance of H. panduraeformis with the desired economic characters of G. hirsutum for rainfed cultivation.The F₁ hybrid showed various meiotic irreguarities and about 40% pollen sterility. Formation of the normal bivalents occurred quite frequently suggesting a close relationship between the parental species. The sterility observed in the hybrid may be due to small structural differences between the chromosomes of the two genera and meiotic abnormalities.     

The transfer of triazine resistance from Brassica napus L. to B. oleracea L. III. First backcross to parental species

Summary Atrazine resistant Brassica napus × B. oleracea F₁ hybrids were backcrossed to both parental species. The backcrosses to B. napus produced seeds in both directions but results were much better when the F₁ hybrid was the pollen parent. Backcrosses to B. oleracea failed completely but BC₁s were rescued by embryo culture both from a tetraploid hybrid (2n = 4x = 37; A₁C₁CC) and sesquidiploid hybrids (2n = 3x = 8; A₁C₁C). Progeny of crosses between the tetraploid hybrid and B. oleracea had between 25 and 28 chromosomes. That of crosses between the sesquidiploid hybrid and B. oleracea had between 21 and 27. A few plants that had chromosome counts outside the expected range may have originated from either diploid parthenogenesis, unreduced gametes or spontaneous chromosome doubling during in vitro culture. Pollen stainability of the BC₁s ranged from 0% to 91.5%. All the BC₁s to B. oleracea were resistant to atrazine.