Transfer of resistance to race 2 of Plasmodiophora brassicae from Brassica napus to cabbage (B. oleracea ssp. capitata). III. First backcross and F2 progenies from interspecific hybrids between B. napus and B. oleracea ssp. capitata

Summary The first backcross and F₂ progenies from triploid F₁ and tetraploid F₁ hybrids between B. napus and 2x and 4x B. oleracea ssp. capitata (cabbage) were studied for their general morphology, resistance to race 2 of the clubroot pathogen, chromosome number and meiotic chromosome behavior. No linkage was apparent between resistance and the major morphological characters. Unreduced gametes played a large part in the successful formation of seed of the B₁ and F₂ progeny. B₁ plants with low chromosome numbers were selected for use in recurrent backcrosses. The potential use of anther culture to extract gametic progenies from resistant B₁ and F₂ plants with higher chromosome numbers was suggested. The presence of homoeologous pairing observed in all the plants is considered advantageous for selecting suitable progeny in later generations.     

Transfer of resistance to race 2 of Plasmodiophora brassicae from Brassica napus to cabbage (B. oleracea var. capitata). II. Meiosis in the interspecific hybrids between B. napus and 2x and 4x cabbage

Summary Meiosis in 14 interspecific F₁ hybrids with three chromosomal levels (triploid, tetraploid, hexaploid; 2n=28, 37 and 55) between Brassica napus L. and 2x and 4x cabbage (B. oleracea var. capitata L.) was studied. The oleracea genome from B. napus maintained close homology with the c genome of cabbage while the campestris genome of B. napus showed partial homology with the c genome contained in the hybrids. Genotypic influence on chromosome pairing was indicated. Structural chromosome differences and spontaneous chromosome breakage and reunion were suggested as causes for the abnormalities which related to the unbalance of the genotypes. The divergence of the genomes of B. napus and B. oleracea and the need for the qualification of the term secondary association were discussed.Contribution No. J. 673, Research Station, Agriculture Canada, St. Jean, Québec.     

The transfer of triazine resistance from Brassica napus L. to B. oleracea L. II. Morphology,fertility and cytology of the F1 hybrid

Summary F₁ hybrids of triazine resistant Brassica napus and triazine susceptible B. oleracea were morphologically intermediate to the parent species. Of 49 hybrids examined, 44 had 28 chromosomes, two had 37, one had 38 and two had 56. The 38-chromosome plant was thought to be a matromorph, the others, A₁C₁C (28), A₁C₁CC (37) or A₁A₁C₁C₁CC (56) type hybrids. Pollen stainability averaged 9.0% in the sesquidiploid, 32.0% in the tetraploids and 89.5% in the hexaploids. All the interspecific hybrids were resistant to 1.0×10^-4 mol L^-1 atrazine. The sesquidiploid hybrids produced gametes with chromosome numbers ranging from 9 to 17 and the tetraploid hybrid gametes had chromosome numbers from 15 to 22. Most hybrids produced self-seed. The partial fertility of these hybrids may permit their backcrossing to one or both parents.     

Transfer of resistance to race 2 of Plasmodiophora Brassicae from Brassica napus to cabbage (B. oleracea var. Capitata). I. Interspecific hybridization between B. napus and B. oleracea var. Capitata

Summary Interspecific hybridization between Brassica napus L. (2n=38, a₁a₁c₁c₁) and B. oleracea var. capitata L. (2x- and 4x-cabbage; 2n=2x=18, cc and 2n=4x=36, cccc) was carried out for the purpose of transferring clubroot disease resistance from the amphidiploid species to cabbage. Nineteen hybrids with three different chromosome levels (2n=28, a₁c₁c; 2n=37, a₁c₁cc and 2n=55, a₁c₁cccc) were obtained. The F₁ plants were mostly intermediate between the two parents but as the number of c genomes in the hybrids increased, the more closely the hybrids resembled the cabbage parent. All F₁ hybrids were resistant when tested against race 2 of Plasmodiophora brassicae wor. The complete dominance of resistance over susceptibility suggested that the gene(s) controlling resistance to this particular race of the clubroot pathogen is probably located on a chromosome of the a genome in Brassica.Contribution No. J654.     

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.     

Crossability and cytology of hybrid progenies in the cross between Brassica campestris and three wild relatives of B. oleracea,B. bourgeaui,B. cretica and B. montana

Summary Crossability and cytology were examined in F₁, F₂, B₁ and hybridsplants of F₁ hybrids of Brassica campestris and three wild relatives of B. oleracea, B. bourgeaui, B. cretica and B. montana, respectively. The F₂ plants were obtained after self-and open pollination of the F₁ hybrids. The B₁ and hybrid plants were produced after the F₁ hybrids backcrosses with B. campestris and crossed with B. napus, respectively. After crossing the F₁ hybrids, many seeds of the F₂, B₁ and hybrid plants were harvested. Multivalent formation was high in the chromsome configuration for the PMCs of F₂, B₁ and hybrid plants, suggesting that crossing over might occur between them. Many different types of aneuploids were obtained in the progenies of the F₂, B₁ and hybrid plants. It is suggested that different types of normal egg cells may be produced by one-by-one or little-by-little chromosome addition. The possibility is discussed of gene transfer from B. bourgeaui, B. cretica and B. montana, to cultivated plants, B. campestris and B. napus.     

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.     

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.     

Inheritance of internode length,plant form,and annual habit in a cross of cabbage and broccoli (Brassica oleracea var. capitata L. and var. italica Plenck.)

Summary When an inbred line of cabbage, Brassica oleracea L. var. capitata L., was crossed with an inbred line of broccoli B. oleraceae var. italica, the F₁ progeny were vigorous late annuals. All F₁ × broccoli backcross plants and 92% of the 3260 F₂ plants were annuals, while 40% of the F₁ × cabbage backcross plants were biennials. Annual habit is thus dominant and controlled by more than a single gene. Number of days to bud appearance in annuals varied continuously, and was primarily additive in inheritance. F₁ data suggested partial dominance for lateness but this was not supported by the F₂. Internode length was also continuous in distribution and primarily additive in inheritance, but with some dominance for short internodes in the F₁. Cabbage head forming ability was recessive and multigenic, with 2% of the F₂ plants forming heads, of which none were of commercial type and about half bolted as annuals. There was a significant chi square association between biennial habit and tendency for cabbage head formation. Clasping habit of terminal leaves was recessive to open leaves, multigenic, and associated with both cabbage heading and biennial habit.Technical Paper 4836, Oregon Agricultural Experiment Station; from an M.S. thesis by the senior author.     

Interspecific cross of Brassica oleracea var. alboglabra and B. napus : effects of growth condition and silique age on the efficiency of hybrid production, and inheritance of erucic acid in the self-pollinated backcross generation

Interspecific hybrids were produced from reciprocal crosses between Brassica napus (2n = 38, AACC) and B. oleracea var. alboglabra (2n = 18, CC) to introgress the zero-erucic acid alleles from B. napus into B. oleracea. The ovule culture embryo rescue technique was applied for production of F₁ plants. The effects of silique age, as measured by days after pollination (DAP), and growth condition (temperature) on the efficiency of this technique was investigated. The greatest numbers of hybrids per pollination were produced under 20°/15°C (day/night) at 16 DAP for B. oleracea (♀) × B. napus crosses, while under 15°/10°C at 14 DAP for B. napus (♀) × B. oleracea crosses. Application of the ovule culture technique also increased the efficiency of BC₁ (F₁ × B. oleracea) hybrid production by 10-fold over in vivo seed set. The segregation of erucic acid alleles in the self-pollinated backcross generation, i.e. in BC₁S₁ seeds, revealed that the gametes of the F₁ and BC₁ plants carrying a greater number of A-genome chromosomes were more viable. This resulted in a significantly greater number of intermediate and a smaller number of high-erucic acid BC₁S₁ seeds.     

Production of 27-, 28-, and 56-chromosome apomictic hybrid derivatives between pearl millet (2n=14) and Pennisetum squamulatum (2n=54)

Summary An interspecific hybridization program designed to transfer gene(s) controlling apomixis from Pennisetum squamulatum Fresen. (2n=6x=54) to induced tetraploid (2n=4x=28) cultivated pearl millet, Pennisetum americanum (L.) Leeke resulted in four offtype plants, two with 27 chromosomes and two with 28 chromosomes. These plants were found among 217 spaced plants established from open-pollinated seed of an apomictic 21-chromosome polyhaploid (2n=21) plant derived from an apomictic interspecific hybrid (2n=41) between tetraploid pearl millet and Pennisetum squamulatum. It appeared that a 21- (or 20-) chromosome unreduced egg from the apomictic polyhaploid united with a 7-chromosome pearl millet (2n=2x=14) gamete to produce a 28- (or 27-) chromosome offspring. Meiotic chromosome behavior was irregular averaging from 3.60 to 4.05 bivalents per microsporocyte in the 27- and 28-chromosome hybrids. The 27- or 28-chromosome hybrids, like the 21-chromosome female parent, shed no pollen, but set from 1.8 to 28 seed per panicle when allowed to outcross with pearl millet. Progeny of the 28-chromosome hybrids were uniform and identical to their respective female parents, indicating that apomixis had been effectively transferred through the egg. In addition, a 56-chromosome plant resulting from chromosome doubling of a 28-chromosome hybrid was identified. Pollen was 68 per cent stainable and the plant averaged 2.3 selfed seeds per panicle. Chromosomes of the 56-chromosome plant paired as bivalents (x=10.67) or associated in multivalents. Three to nine chromosomes remained unpaired at metaphase I. Multiple four-nucleate embryo sacs indicated the 56-chromosome hybrid was an obligate apomict. The production of 27-, 28-, and 56-chromosome hybrid derivatives were the results of interspecific hybridization, haploidization, fertilization of unreduced apomictic eggs, and spontaneous chromosome doubling. These mechanisms resulted in new unique genome combinations between x=7 and x=9 Pennisetum species.     

Inheritance of internal pigmentation in green cabbage (Brassica oleracea var. Capitata L.)

Summary Internal anthocyanin pigmentation (IP) in otherwise normally green cabbage occurs in a number of Oregon State University breeding lines. Extracted pigment, tested for spectral absorption and for color reactions with lead acetate and aluminium chloride, was similar but not necessarily identical to pigment extracted from red cabbage cultivar Redman. When IP line R52 was crossed with normal green line C70, the F₁, F₂ and backcross progenies indicated that IP at the intensity found in R52 was determined by a single factor in homozygous condition, with intermediate levels of IP expressed by the heterozygous genotypes. Modifying factors also appear to influence the level of IP. In the cross R52 (IP)×R51 (normal green), expression of IP in the F₁ was much reduced. The F₂ failed to fit the expected 3 IP: 1 green ratio due to an excess of green plants, but instead, closely fit a 9:7 ratio. This may have resulted from incomplete expression of IP because of modifiers, rather than from the effects of a second major gene. An allele at the A (anthocyanin) locus of B. oleracea is tentatively proposed and designated A ^IP or a ^IP pending further identification.Oregon Agricultural Experiment Station Technical Paper No. 4690.     

Inheritance of seed colour in Brassica campestris L. and breeding for yellow-seeded B. napus L.

Summary Seed colour inheritance was studied in five yellow-seeded and one black-seeded B. campestris accessions. Diallel crosses between the yellow-seeded types indicated that the four var. yellow sarson accessions of Indian origin had the same genotype for seed colour but were different from the Swedish yellow-seeded breeding line. Black seed colour was dominant over yellow. The segregation patterns for seed colour in F₂ (Including reciprocals) and BC₁ (backcross of F₁ to the yellow-seeded parent) indicated that the black seed colour was conditioned by a single dominant gene. Seed colour was mainly controlled by the maternal genotype but influenced by the interplay between the maternal and endosperm and/or embryonic genotypes. For developing yellow-seeded B. napus genotypes, resynthesized B. napus lines containing genes for yellow seed (Chen et al., 1988) were crossed with B. napus of yellow/brown seeds, or with yellow-seeded B. carinata. Yellow-seeded F₂ plants were found in the crosses that involved the B. napus breeding line. However, this yellow-seeded character did not breed true up to F₄. Crosses between a yellow-seeded F₃ plant and a monogenomically controlled black-seeded B. napus line of resynthesized origin revealed that the black-seeded trait in the B. alboglabra genome was possibly governed by two independently dominant genes with duplicated effect. Crossability between the resynthesized B. napus lines as female and B. carinata as male was fairly high. The sterility of the F₁ plants prevented further breeding progress for developing yellow-seeded B. napus by this strategy.     

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.     

Giemsa N-banding pattern in cabbage and Chinese kale

Summary In cabbage (Brassica oleracea var. capitata) and Chinese kale (B. oleracea alboglabra) four types of N-bands can be distinguished: pericentromeric, telomeric (terminal), intercalary and satellite bands. Typical NOR bands were not observed. The pericentromeric bands appear at the pericentric regions, possibly even at the centromeres of all chromosomes. Telomeric bands are observed on the short arms of chromosomes 1,5 and 6 in cabbage and chromosomes 1 and 5 in Chinese kale. Intercalary bands stained weakly in the long arms of chromosome 3 in cabbage and chromosome 2 in Chinese kale. Satellite bands cover the entire satellites in both Brassica species. The N-banding pattern is very similar in appearance to the C-banding pattern in both species and much more convenient to apply.     

The allelic relationship between A IP(internal pigmentation) and A RC (red cabbage) in Brassica oleracea var. capitata L.

Summary Internal anthocyanin pigmentation (IP), previously reported to be controlled by a single gene dominant to aa of normal green cabbage, was studied further to determine its relationship to A ^RC A ^RC of red cabbage. When IP line R52 was crossed with an early red cabbage line. F₁ heads were pigmented throughout, but pigment intensity was intermediate. Subjective classification of F₂ plants by pigmentation intensity and distribution scores gave a ratio of 1 intense red throughout (red cabbage):2 medium red throughout:1 medium to light red, restricted to the central portion of the head (IP). The genotypes A ^RC A ^RC: A ^RC A ^IP: A ^IP A ^IP. respectively, are proposed to explain these 3 phenotypic classes. F₂ progenies contained no normal green plants, supporting the conclusion that A ^IP and A ^RC are alleles.Technical Paper 7052, Oregon Agricultural Experiment Station.     

An S-allele survey of cabbage (Brassica oleracea var. Capitata)

Summary A total of 31 S-alleles was found in a survey of 197 cabbage plants representing 11 cultivars of diverse type. Most of these S-alleles also occurred in either kale or Brussels sprouts, but five of them have not been found previously and apparently occur only in cabbage. A more detailed study of five cultivars of spring cabbage showed only 12 S-alleles in all, with 6–10 S-alleles in four older cultivars and only 3 S-alleles in the newer more highly selected cultivar. S2 was by far the commonest S-allele, as it is in B. oleracea as a whole. The highly recessive alleles S5 and S15 were not particularly common in cabbage and this may partly explain why the sib problem in F₁ hybrids is apparently less in cabbage than in Brussels sprouts. Three cases were found in which an S-allele was completely recessive in both the stigma and the pollen. The problems for the breeder created by this rather unusual situation are discussed.     

Intergeneric hybrid between Brassica napus and Diplotaxis harra through ovary culture and the cytogenetic analysis of their progenies

Brassica napus (2n = 38) and Diplotaxis harra (2n = 26) were used to investigate gene transfer from D. harra to B. napus. Intergeneric F₁ hybrids (dihaploid 2n = 32 chromosomes) were obtained through ovary culture. The chromosome associations in the first meiotic division was (0–2)_III + (2–10)_II + (12–28)_I. Many seeds were harvested in the F₁ hybrid after backcrossing with B. napus, and from open pollination of the F₁ hybrid. Somatic chromosome numbers of BC₁ and hybrid plants varied from 2n = 26 to 52. In the first meiotic division, high frequencies of bivalent association and relatively low pollen fertility were observed. BC₂ plants generated from the BC₁ plants with 2n = 38 chromosomes, 69.6% showed 2n = 38 chromosomes. Many aneuploids with addition and deletion of chromosomes were also obtained. A bridge plant between B. napus and D. harra with 2n = 32 chromosomes should be valuable material for the breeding of brassica crops.     

Stabilization of new types of diploids (2n=22, 24) through selfing of aneuploids (2n=21, 22) derived from crossing of sesquidiploids (2n=29, AAC) and Brassica campestris (2n=20 AA)

Summary Aneuploids with 2n=21 and 2n=22 derived from crossing of sesquidiploids (2n=29, AAC) and Brassica campestris (2n=20, AA) were selfed successively in order to follow the changes in chromosome number of the progenies for three consecutive generations. Progenies with 2n=22, 23 and 24 obtained after selfing of S₀ generation and the succeeding S₁, S₂ and S₃ generations were analyzed in terms of pollen stainability, % seed set as well as cytogenetically based on meiotic behaviour with the aim of determining the possibility of addition of one or more alien chromosomes into n=10 species which may lead to differentiation of single or plural disomic addition lines. The generation of aneuploids with 2n=21 progressed in such a way that most plants seem to revert to the 2n=20 chromosome number of B. campestris after selfing. From 2n=22 aneuploids, however, the succeeding progenies showed high frequency of plants with two additional chromosomes which accounted for 50.6% and 52.9% of total S₃ progenies via 2n=22 and 2n=24 S₂ generations, respectively. The meiotic behaviour of these progenies indicated evidence for a rule governing the frequency distribution of chromosome number among these addition lines and high possibility to breed such disomic plants with 2n=22. A method of selecting stable aneuploids was suggested in addition to the possible role of pollination biology at various processes of such breeding program.     

Evidence for the production of unreduced gametes by tetraploid Rosa hybrida L.

The objective of this study was to determine the mode of inheritance of field resistance to downy mildew (Peronospora parasitica (Pers. ex Fr.) Fr.) in broccoli (Brassica oleracea var.italica) at the adult plant stage. The F₁, F₂ and F₃ progeny of resistant and susceptible plants of broccoli were tested in the field under natural infection, in central Portugal, from August to December in two successive years. The plants were evaluated for resistance to downy mildew at maturity using a five-class scale of increasing susceptibility to the disease, which took into account the number of infected leaves and the size of the sporulating lesions. The F₁ was completely resistant, the F₂ segregated a clear 3 resistant: 1susceptible and the F₃ confirmed the F₂ segregation, which suggests a dominant character controlled by a single locus. This resistance has good potencial for direct use in commercial broccoli breeding or for transfer to other Brassica vegetables. This revised version was published online in August 2006 with corrections to the Cover Date.