Hybridizing <Emphasis Type="Italic">Brassica rapa</Emphasis> with wild crucifers <Emphasis Type="Italic">Diplotaxis erucoides</Emphasis> and <Emphasis Type="Italic">Brassica maurorum</Emphasis>

Two wide hybrids, Diplotaxis erucoides (2n = 14) × Brassica rapa (2n = 20) and B. maurorum (2n = 20) × B. rapa, were developed using the sequential ovary–ovule culture. Reciprocal crosses failed, possibly as a consequence of strong unilateral incompatibility. The F ₁ hybrids in each combination were completely male sterile and morphologically intermediate to the respective parents. DNA marker polymorphism and chromosome counts confirmed their hybrid nature. High frequency of bivalents in the F ₁ and the presence of trivalents/quadrivalents in the derived amphiploids suggested genomic duplications and homoeology of the parental genomes. Up to three homoeologous pairs between the D. erucoides (D^eD^e) and B. rapa (AA) genomes, and one between B. maurorum (B^mB^m) and B. rapa genomes were observed. Successful synthesis of the F ₁ hybrids and amphiploids of B. rapa with D. erucoides and B. maurorum, and allosyndetic chromosome pairing are expected to permit introgressions of desirable loci into the cultivated Brassica germplasm, especially for resistance to Alternaria brassicae and Albugo candida.     

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.     

Phenotypic and seed structural comparison in hybrids of different Brassica rapa and B. oleracea

Brassica napus is an important oil species with short history and narrow genetic background. Interspecific hybrids from crosses between B. oleracea and different B. rapa were obtained. We found the hybrids with white petal resembling B. oleracea, the flavonoid and phenolic content decreased in hybrids, agreeing with the expressional changes of flavonoid biosynthesis genes. Seed coat of hybrids resembled diploid parents, or partly resembled to each parent with a clear outline. The palisade layer in hybrids was thicker than parents, with similar pigment accumulation as B. oleracea but more than B. rapa. Differentially sized protein bodies (PBs) were found in hybrids. The radical and inner cotyledon of all hybrids were identified with larger but less PBs than parents. The average size of PBs in outer cotyledon of resynthesized B. napus was also larger than parents, but the number of PBs was not significantly reduced. The phenotypic and seed structural variations after polyploidization of B. napus would be interesting for genetic broadening and breeding of rapeseed.     

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.     

Meiotic chromosome pairing in intergeneric hybrids of colchicaceous ornamentals revealed by genomic in situ hybridization (GISH)

Diploid and triploid intergeneric hybrids obtained by crosses among Gloriosa superba ‘Lutea’ (2n = 2x = 22), G. ‘Marron Gold’ (2n = 4x = 44), Littonia modesta (2n = 2x = 22), and Sandersonia aurantiaca (2n = 2x = 24) were analyzed for their meiotic chromosome pairing in pollen mother cells by genomic in situ hybridization (GISH) with digoxigenin-labeled total DNA of one parent as probe. Chromosomes from each parent could be clearly distinguished in pollen mother cells of all the five intergeneric hybrids by GISH. For three diploid hybrids, L. modesta × G. superba ‘Lutea’ (2n = 2x = 22), L. modesta × S. aurantiaca (2n = 2x = 23) and S. aurantiaca × G. superba ‘Lutea’ (2n = 2x = 23), 0.04?0.27 autosyndetic bivalents (intragenomic pairing of non-homologous chromosomes) and 0.13?0.36 allosyndetic bivalents (intergenomic chromosome pairing) were observed per pollen mother cell, indicating that there are some homologous chromosomal regions within each genome and among the genomes of Gloriosa, Littonia and Sandersonia. Differences in the average number of allosyndetic bivalents per pollen mother cell among different genome combinations may reflect the evolutionary distances among the three genera, and Gloriosa and Littonia may be closely related to each other, while Sandersonia may have relatively distant relationships with Gloriosa and Littonia. For two triploid hybrids, L. modesta × G. ‘Marron Gold’ (2n = 3x = 33) and S. aurantiaca × G. ‘Marron Gold’ (2n = 3x = 34), no allosyndetic bivalents were observed. Based on the results obtained in the present study, possible utilization of the diploid and triploid intergeneric hybrids for further breeding of colchicaceous ornamentals is discussed.     

Novel interspecific hybridization between sweetpotato (<Emphasis Type="Italic">Ipomoea batatas</Emphasis> (L.) Lam.) and its two diploid wild relatives

Interspecific hybridization is an important approach to broaden the genetic base and create novel plant forms in breeding programs. However, interspecific hybridization in Ipomoea is very difficult due to the cross incompatibility. Here we report two novel interspecific F₁ hybrids between I. batatas (L.) Lam. (2n = 6x = 90) and two wild species, I. grandifolia (2n = 2x = 30) and I. purpurea (2n = 2x = 30). Hybridization was stimulated by applying plant growth hormones. Morphological, molecular and cytological tests were conducted to confirm their hybridity. We found that the two hybrids were quite distinctive in leaf color and morphology, and yielded intermediate sizes of storage roots compared to their respective parents. Inter-simple sequence repeat analysis showed that the unique DNA bands from the wild parents could be detected in these two hybrids. The cluster analysis also showed that the two F₁ hybrids were closer to I. batatas in phylogeny relationship. The number of chromosomes of the two hybrids was both 60, indicating that the hybrids were tetraploid. The meiotic configuration analysis of the H₁ of I. batatas × I. grandifolia revealed the occurrence of 17.58I + 14.28II + 1.36III + 2.48IV at metaphase I in average chromosome association per pollen mother cells (PMCs), 4.26I + 18.32II + 2.56III + 3.12IV was average meiotic configuration in the H₂ of I. batatas × I. purpurea. Both hybrids appeared to be polyads and multi-microcytes at tetrad phase and differed in their pollen fertility.     

Genetic effects on biomass yield in interspecific hybrids between Brassica napus and B. rapa

This study was conducted to estimate the genetic effects on biomass yield in the interspecific hybrids between Brassica napus and B. rapa, and to evaluate the relationship between parental genetic diversity and its effect on biomass yield of interspecific hybrids. Six cultivars and lines of oilseed B. napus and 20 cultivars of oilseed B. rapa from different regions of the world were chosen to produce interspecific hybrids using NC design II. Obvious genetic differences between B. rapa and B. napus were detected by RFLP. In addition, Chinese B. rapa and European B. rapa were shown genetically differences. Plant biomass yield from these interspecific hybrids were measured at the end of flowering period. Significant differences were detected among general combining ability (GCA) effects over two years and specific combining ability (SCA) effects differences were detected in 2000. The ratios of mean squares, (σ² _GCA(f) + σ² _GCA(m)) / (σ² _GCA(f) + σ² _GCA(m) + σ² _SCA), were 89% and 88% in 1999 and 2000, respectively. This indicates that both additive effects and non-additive effects contributed to the biomass yield of interspecific hybrids and the former played more important role. Some European B. rapa had significant negative GCA effects while many of Chinese B. rapa had significant positive GCA effects, indicating that Chinese B. rapa may be a valuable source for transferring favorable genes of biomass yield to B. napus. Significant positive correlation between parental genetic distance and biomass yield of interspecific hybrids implies that larger genetic distance results in higher biomass yield for the interspecific hybrids. A way to utilize interspecific heterosis for seed yield was discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Introgression of Resistance to Shattering in Brassica napus from Brassica juncea through Non-Homologous Recombination

Genetic information conffering non- shattering of siliques has been introgressed in rapeseed (Brassica napus) following; interspecific hybridization between Brassica juncea and B. napus. A reconstituted B. napus plant with complete non-dehiscence of its fruits was isolated in the BC-, generation. This plant had normal meiosis and formed 19 bivalents. Its seed fertility, however, was low (23 percent). It is suggested that the gene(s) for shattering-resistance were incorporated into a B. oleracea chromosome following allosyndetic; chromosome pairing and. segmental exchange between B. nigra and B. oleracea chromosomes in the initial interspecific AABC hybrid.     

Variations in chlorosis and potential usefulness of alloplasmic Brassica rapa with the cytoplasm of male sterile Brassica juncea

To select superior seed parents for vegetable hybrid seed production, we conducted interspecific crosses between male sterile Brassica juncea (2n = 36, AABB) and eight inbred lines of Brassica rapa (2n = 20, AA). Alloplasmic lines of B. rapa with the cytoplasm of B. juncea were developed from B. juncea × B. rapa hybrids by repeated backcrossing using B. rapa as the recurrent male parent until the BC₃ generation. Seed fertility, male sterility and chlorophyll content were investigated in these plants cultivated under four different temperature conditions (5, 10, 12 and 20°C). At 10°C, the alloplasmic lines of B. rapa with the cytoplasm of B. juncea were male sterile with partly chlorotic leaves. The alloplasmic B. rapa had lower chlorophyll a, chlorophyll b and carotenoid contents than those of the original B. rapa. The leaves recovered from chlorosis when the plants were cultivated at 20°C. An alloplasmic line of B. rapa (A6) is available as a seed parent for vegetable hybrid seed production and contributes seed fertility, slight chlorosis and stable male sterility.     

The Effect of A Genome Substitution on the Resistance of Brassica napus to Infection by Leptosphaeria maculans

Six accessions belonging to four subspecies of Brassica rapa, including three accessions of B. rapa subsp. sylvestris, were crossed with B. oleracea subsp. alboglabra in order to develop a series of synthetic B. napus lines with a common C genome but contrasting A genomes. Different A genomes had significant effects on the efficiency of B. napus resynthesis and the sexual compatibility of the synthetic lines with oilseed rape cultivars. The synthetic lines were used to investigate the effect of A genome substitution on the resistance of B. napus to infection by Leptosphaeria maculans, and to explore the potential for the use of wild forms of B. rapa in oilseed rape breeding programmes. Synthetic lines derived from two wild accessions of B. rapa, and their F₁ hybrids with oilseed rape cultivars, expressed high levels of resistance to L. maculans in glasshouse experiments. One of these lines also expressed high levels of resistance in field experiments in England and Australia when exposed to a genetically diverse pathogen population. All other synthetic lines and cultivars were highly susceptible in both glasshouse and field experiments. F₁ hybrids between oilseed rape cultivars and synthetic lines derived from B. rapa subsp. chinensis were significantly more susceptible than either parent.     

Meiotic nondisjunction in resynthesized <Emphasis Type="Italic">Brassica napus</Emphasis> and generation of aneuploids through microspore culture and their characterization

Meiotic nondisjunction during microsporegenesis can lead to aneuploid gametes formation and reduced pollen fertility in plants. This paper reports the prevalence of meiosis I nondisjunction in a resynthesized Brassica napus (AACC, 2n = 38) and its use for aneuploid production. Meiosis in the amphidiploids was characterized by high frequencies of univalents and multivalents per PMC at diakinesis/metaphase I and notably unbalanced chromosome segregations at anaphase I (AI). In all the plants observed, 18.95–44.3% of PMCs exhibited a segregation of 18:20 (n − 1:n + 1) at AI which was caused by nondisjunction of one bivalent or the distribution of two homologous univalents to the same pole. Meiosis proceeded normally after AI then, thus led to the formation of viable n − 1 and n + 1 gametes and high pollen fertility of these plants. Microspore culture was subsequently carried out using these plants in an attempt to isolate Brassica nullisomics. Four nullisomics (2n = 36), two nullihaploids (2n = 18) and one tetrasomic haploid (2n = 20) were identified cytologically and characterized morphologicaly and physiologically. Amplified fragment length polymorphism (AFLP) survey suggested that of the six nullisomics/nullihaploids, one nullihaploids lost one A-genome chromosome and the other five lost C genome chromosome(s). Altogether, different C-genome chromosomes were thought to have been lost in the nullisomics/nullihaploids. The mechanisms underlying the meiotic abnormalities and the implications of these B. napus nullisomics are discussed.     

Hybrids of <Emphasis Type="Italic">Avena sativa</Emphasis> with two diploid wild oats (CIav6956) and (CIav7233) resistant to crown rust

Two diploid accessions of wild oat, CIav6956 and CIav7233, were identified as carrying seedling resistance to oat crown rust (caused by Puccinia coronata f. sp. avenae; Pca). Two vigorous interploidy F₁ hybrids were generated from crosses involving the hexaploid oat cultivar Wintaroo and the diploid oat Avena strigosa Schreb. accession CIav6956. An additional interploidy F₁ hybrid, designated “F1-Aa1”, was produced from a cross of Wintaroo and the diploid oat accession CIav7233. All three hybrids were more vigorous and taller than the cultivated parent Wintaroo. The three F₁ hybrids contained full chromosome complements from both parents (2n = 4x = 28), but no seeds were obtained when the three F₁ hybrids were selfed. Meiotic analyses of the hybrids indicated that they exhibited a high degree of inter-genome and intra-genome pairing. Trivalent configurations were detected in 95–96% of meiotic cells and a minimum of three bivalents was present in all cells. An average chiasma frequency of 7.2–7.9 per cell was observed for the three F₁ hybrids. A fourth F₁ hybrid was subsequently generated from a cross between the diploid oat accession CIav7233 and Wintaroo. One octaploid (2n = 8x = 56) was generated from this hybrid and progeny were resistant to two Pca races. The chromosome number of the octaploid progeny varied between 51 and 54 chromosomes. Development of a chromosome addition line(s) with the crown rust resistance should be possible from these partial-octaploids.     

Interspecific hybrids between <Emphasis Type="Italic">Dendranthema morifolium</Emphasis> (Ramat.) Kitamura and <Emphasis Type="Italic">D. nankingense</Emphasis> (Nakai) Tzvel. achieved using ovary rescue and their cold tolerance characteristics

Ovary rescue was employed to create six interspecific hybrids from the cross between Dendranthema morifolium (Ramat.) Kitamura ‘rm20-12’ (2n = 54) and its wild diploid relative D. nankingense (Nakai) Tzvel. (2n = 18). The morphology of the hybrids differed from that of either parent. The leaf length and width of all three D. morifolium × D. nankingense hybrids exceeded that of the parents, as did the plant height of two and the inflorescence diameter of another of the hybrids. One of the reciprocal hybrids was heterotic for leaf length and width. All the hybrids bore yellow flowers. The cold tolerance of five hybrids was significantly superior to that of their D. morifolium parent. Interspecific hybridization clearly provides an effective means of cultivar improvement in chrysanthemum.     

Homoeological relationships between the f chromosome of Brassica rapa and the e chromosome of Brassica oleracea

Eight plants of the putative double monosomic addition line (DMAL, 2n= 20) were developed by crossing a monosomic chromosome addition line of radish [f(A)‐type monosomic addition line (MAL) (2n= 19)] carrying the f chromosome of Brassica rapa (2n= 20, AA) with another [e(C)‐type MAL (2n= 19)] having the echromosome of Brassica oleracea (2n= 18, CC). The homoeological relationships between the two alien chromosomes were investigated by morphological, cytogenetic and random amplified polymorphic DNA (RAPD) analysis. Seventeen morphological traits that were not present in the radish cv. ‘Shogoin’ were observed in both MALs and these traits were substantially exhibited in DMAL plants. At the first metaphase of pollen mother cells (PMCs), the two parental MALs showed a chromosome configuration of 9II +1I, demonstrating impossibility of recombination between the R and the added chromosomes. The DMALs formed 10II in approximately 73% of PMCs, with one bivalent showing loose pairing between two chromosomes differing in size. In an attempt to identify the two MALs by RAPD‐specific markers using 26 selected random primers, 13 and 20 bands were specific for the f(A)‐type and the e(C)‐type MALs, respectively; 12 bands were common to both MALs (26.7%). In conclusion, the f chromosome of B. rapa is homoeologous to the e chromosome of B. oleracea. The genetic domain (genes) for 17 morphological traits are linked to each homoeologous chromosome bearing 27% of the corresponding RAPD markers.     

A cytogenetic study of the progenies of hybrids between Brassica napus and Brassica oleracea, Brassica bourgeaui, Brassica cretica and Brassica montana

In this cytogenetic study the progeny of all crosses were investigated in F₁, F₂ and backcross (BC₁) hybrids. Brassica napus and F₁ hybrids between B. napus and B. oleracea, and between B. napus and three wild relatives of B. oleracea (B. bourgeaui, B. cretica and B. montana). Each of the wild relatives has 18 somatic chromosomes. Interspecific F₁ hybrids were obtained through ovary culture mean. These had 28 and 37 chromosomes and their mean pollen fertility was 10.7% and 93.0%, respectively. Many F₂ and BC₁ seeds were harvested from the F₁ hybrids with 37 chromosomes after self‐pollination and open pollination of the F₁ hybrids, and backcrossing with B. napus. Many aneuploids were obtained in the F₂ and BC₁ plants. It is evident from these investigations that the F₁ hybrids may serve as bridge plants to improve B. napus and other Brassica crops.     

Morphological,cytological and reproductive characterization of tri-species hybrids (GOS) between <Emphasis Type="Italic">Pennisetum glaucum</Emphasis>, <Emphasis Type="Italic">P. orientale</Emphasis> and <Emphasis Type="Italic">P. squamulatum</Emphasis>

New tri-species hybrids (GOS) in the genus Pennisetum involving the cultivated species pearl millet (P. glaucum L.) and two wild species, viz. P. squamulatum Fresen and P. orientale L. C. Rich, are reported. Six hybrid plants were recovered after crossing a backcross hybrid (2n = 3x = 23, GGO) between P. glaucum (2n = 2x = 14, GG) and P. orientale (2n = 2x = 18, OO) with F₁s (2n = 6x = 42, GGSSSS) between P. glaucum (2n = 4x = 28, GGGG) and P. squamulatum (2n = 8x = 56, SSSSSSSS). The hybrids were perennial, morphologically intermediate to their parents, and represented characters from the three contributing species. The hybrids contained 2n = 44 chromosomes (GGGSSO) representing 21, 14 and nine chromosomes from P. glaucum, P. squamulatum and P. orientale, respectively. Meiotic and flow-cytometric analysis suggested origin of these hybrids from unreduced female and reduced male gametes. Average chromosome configuration (8.42_I + 14.32_II + 1.62_III + 0.52_IV) at Meiosis showed limited inter-genomic pairing indicating absence of significant homology between the three genomes. The hybrids were male sterile (except one) and highly aposporous. P. orientale was identified to induce apospory in hybrid background with P. glaucum at diploid and above levels, though it was quantitatively affected by genomic doses from sexual parent. A case of inducible and recurrent apospory is presented whereby a transition from Polygonum-type sexual embryo-sacs to Panicum-type aposporous embryo-sacs was observed in diploid interspecific hybrids. Results supported independent origin and partitioning of the three apomixis-components (apomeiosis, parthenogenesis, and functional endosperm development), reported for the first time in Pennisetum. Potential utilization of GOS hybrids in understanding genome interactions involved in complex traits, such as perenniality and apomixis, is discussed.     

A new method for producing allohexaploid Brassica through unreduced gametes

We trialled a two-step method of producing allohexaploid Brassica with three genomes (A, B, and C) derived from pair-wise crossing among three allotetraploid Brassica species. In the first step, the three allotetraploid Brassica species (Brassica juncea, A^jA^jB^jB^j; Brassica napus, AⁿAⁿCⁿCⁿ; and Brassica carinata, B^cB^cC^cC^c) were intercrossed in pairs to produce unbalanced trigenomic hybrids: A^jAⁿB^jCⁿ, B^jB^cA^jC^c and CⁿC^cAⁿB^c. In the second step, these hybrids were crossed with the complementary allotetraploid parent, that is, A^jAⁿB^jCⁿ × B^cB^cC^cC^c (B. carinata), B^jB^cA^jC^c × AⁿAⁿCⁿCⁿ (B. napus) and CⁿC^cAⁿB^c × A^jA^jB^jB^j (B. juncea). We hypothesised that the unbalanced trigenomic hybrids would produce high levels of unreduced gametes with the same genome composition as the hybrid. These unreduced gametes would unite with reduced gametes from the complementary allotetraploids to form allohexaploid Brassica progeny (A^jAⁿB^cB^jC^cCⁿ). From 112 s generation interspecific progeny, two progeny were shown by locus-specific simple sequence repeat markers to be near-allohexaploids derived from an unreduced gamete from CⁿC^cAⁿB^c and a reduced gamete from B. juncea (A^jB^j). One of these plants was highly self-fertile, had 50 chromosomes, and inherited patterns of marker alleles (A^jAⁿB^cB^jC^cCⁿ) that were predicted from first division restitution at meiosis in the CⁿC^cAⁿB^c parent. The second near-allohexaploid had 60 chromosomes, was sterile, and inherited patterns of marker alleles that indicated second division restitution in the CⁿC^cAⁿB^c parent. Trigenomic hybrid Brassica produced by these methods will be valuable bridges to move alleles between allotetraploid species, and may contribute useful meiotic stability alleles to a future allohexaploid species.     

Nuclear genetic control of variation in simazine tolerance in oilseed brassicas

Summary Brassica napus is a natural allotetraploid derived from the diploid species B. rapa L. (syn. campestris L.) and B. oleracea L. Somatic hybrids synthesized from highly heterozygous lines of these two diploid species were evaluated for fertility. The hybrids were obtained from two fusion experiments which differed in the B. rapa full-sibling parent used as the source of protoplasts. Both B. rapa siblings were lelf-incompatible (SI) yet contained different S-alleles; the B. oleracea species parent was self-compatible (SC). Eight tetraploid hybrids examined had very high female and male fertility; eight hybrids with higher ploidy had low fertility. Hybrids derived from one B. rapa sibling were self-incompatible, whereas those derived from the other B. rapa sibling were fully self-compatible. These data suggest that the different S-alleles of each B. rapa sibling displayed varying penetrance relative to the SC of the B. oleracea parent when combined in B. napus.Abbreviations SC self-compatibility - SI self-incompatibility     

Disease response of resynthesized Brassica napus L. lines carrying different combinations of resistance to Plasmodiophora brassicae Wor.

Resistance responses of resynthesized Brassica napus lines to infection with Plasmodiophora brassicae were investigated. Lines that were derived from interspecific crosses between clubroot-resistant B. rapa and resistant B. oleracea exhibited very broad and effective resistance in both greenhouse and field tests. When clubroot resistance was introduced into resynthesized lines from the B. oleracea parent only, the plants were mainly susceptible. Interspecific hybrids from the most resistant parental genotypes, i.e. B. campestris ECD-04 and the B. oleracea cultivars ECD-15 or ‘Bohmerwaldkohf’, were used to initiate a B. napus resistance-breeding programme. These artificial rapeseed lines were resistant to isolates that were virulent on all B. napus differential lines and/or parental lines. Preliminary segregation analysis suggests that their resistance is due to at least two dominant and unlinked genes. In some cases progenies from selfed resynthesized plants exhibited resistance reactions that differed from those of the parental hybrid plant; this may have been the result of cytological instability.     

Improving ovary and embryo culture techniques for efficient resynthesis of <Emphasis Type="Italic">Brassica napus</Emphasis> from reciprocal crosses between yellow-seeded diploids <Emphasis Type="Italic">B. rapa</Emphasis> and <Emphasis Type="Italic">B. oleracea</Emphasis>

The primary aim of this study was to optimize in vitro culture protocols to establish an efficient reproducible culture system for different Brassica interspecific crosses, and to synthesize yellow-seeded Brassica napus (AACC) for breeding and genetical studies. Reciprocal crosses were carried out between three B. rapa L. ssp. oleifera varieties (AA) and five accessions of B. oleracea var. acephala (CC). All the parental lines were yellow-seeded except one accession of B. oleracea. Hybrids were obtained through either ovary culture from crosses B. rapa × B. oleracea, or embryo culture from crosses B. oleracea × B. rapa. A higher rate of hybrid production was recorded when ovaries were cultured at 4–7 days after pollination (DAP). Of different culture media, medium E (MS with half strength macronutrients) showed good response for ovaries from all the crosses, the highest rate of hybrid production reaching 45% in B. rapa (1151) × B. oleracea (T₂). In embryo culture, the hybrid rate was significantly enhanced at 16–18 DAP, up to 48.1% in B. oleracea (T₃) × B. rapa (JB₂). The combinations of optimal DAP for excision and media components increased recovery of hybrids for ovary and embryo culture, and constituted an improved technique for B. rapa × B. oleracea crosses. In addition, yellow seeds were obtained from progenies of two crosses, indicating the feasibility of developing yellow-seeded B. napus through the hybridization between yellow-seeded diploids B. rapa and B. oleracea var. acephala.