Artificial synthesis of Brassica juncea Coss

Commonly cultivated mustard, Brassica juncea Coss, is an amphidiploid having in its genetic system the full 20-chromosome A genome (Brassica campestris) and the 16-chromosome B genome (Brassica nigra). Considerable natural variability exists under the A genome. These variations have been exploited for the artificial synthesis of B. juncea in order to breed improved mustard. The different combinations were studied both in their F₁'s and advanced amphidiploid generations in respect of their morphology, meiotic behaviour and fertility. Amphidiploids from leafy and rapiferous groups were generally bushy having arboreal habit. Some combinations from the leafy group result in types with luxuriant vegetative growth and can be used for fodder purposes. The amphiploids of ssp. rapifera did not give a swollen and enlarged root like the mother parent. None of the combinations from these two groups was promising in respect of oil and seed yield. Amphidiploids from the oleiferous group were both high seed and oil yielders and thus provide evidence that it formed one of the constituent parental species in the formation of oil yielding B. juncea.     

Investigations on the artificial synthesis of amphidiploids of Brassica tournefortii Gouan with the other elementary species of Brassica. I. Genomic relationships

With the object of studying the genomic relationships of Brassica tournefortii Gouan with the other elementary species of Brassica viz. B. campestris (2n=20, A genome), B. oleracea (2n=18, C genome) and B. nigra (2n=16, B genome), it has been hybridized with them. The percentage of F₁ hybrids formed, their morphology and meiotic behaviour have been described. Based upon crossability relationships and meiotic pairing in the F₁ hybrids, it is inferred that the D genome of B. tournefortii is more closely related to the A genome than to the B and C genomes. It may have been derived from the A genome which likewise shows a strong genetic isolation from B and C. The species has developed a strong genetic barrier in the course of its evolution and shows little crossability, high hybrid sterility and no gene flow with any of the other elementary species. The fact that it has not formed any natural amphidiploids with the elementary species which otherwise are formed in all combinations, is more evidence that it originated more recently than the A genome. It is presumed that B. tournefortii, being more distantly related to B. nigra than to other elementary species, may form stable artificial aphid resistant amphidiploids with the former.     

GENOME ANALYSIS OF AGROPYRON REPENS × AGROPYRON CRISTATUM SYNTHETIC HYBRIDS

Hexaploid A. repens, 2n = 42, and diploid A. cristatum, 2n = 14, were hybridized and gave rise to two 28-chromosome reciprocal hybrids. Approximately 1% of hand-emasculated florets of both parent species produced viable hybrid seed following controlled pollination. Early embryo abortion prevented greater hybrid seed set on A. repens, whereas failure of fertilization appeared to be the major cause of poor hybrid seed set on A. cristatum. Reciprocal differences in hybrid vegetative and spike morphology were striking. The A. repens × A. cristatum hybrid was vigorous, highly rhizomatous, and bore abundant spikes whose morphology was intermediate between that of the parent species. A. cristatum × A. repens hybrids were weak, non-rhizomatous with frequently-malformed spikes. Mean chromosome associations of 0.10 I, 20.10 II, and 0.43 IV were observed in 134 metaphase-I cells of A. repens. Subsequent meiotic stages were regular except for occasional laggards and bridges at anaphase I and II. Metaphase-I chromosome associations averaged 0.07 I and 6.97 II in 124 A. cristatum cells. Chromosome pairing in the hybrids was highly variable and averaged 11.45 I, 7.58 II, 0.44 III, and 0.02 IV per cell in 187 cells interpreted. From 5 to 14 laggards appeared in every hybrid cell at anaphase I. Bridges were observed in approximately 25% of the anaphase-I cells. Similar irregularities were observed at anaphase II. Pollen viability was estimated as 3%, and the hybrids failed to set viable seed. On the basis of chromosome pairing in the species itself and in the hybrids, A. repens was designated as a segmental autoallohexaploid with a genome formula of the type A₁A₁A₂A₂BB. Although A. repens and A. cristatum chromosomes paired occasionally, the genomes of the 2 species were essentially non-homologous. Some of the interpretational difficulties of genome analysis were discussed.     

Trigenomic Bridges for Brassica Improvement

We introduce and review Brassica crop improvement via trigenomic bridges. Six economically important Brassica species share three major genomes (A, B, and C), which are arranged in diploid (AA, BB, and CC) and allotetraploid (AABB, AACC, and BBCC) species in the classical triangle of U. Trigenomic bridges are Brassica interspecific hybrid plants that contain the three genomes in various combinations, either triploid (ABC), unbalanced tetraploid (e.g., AABC), pentaploid (e.g., AABCC) or hexaploid (AABBCC). Through trigenomic bridges, Brassica breeders can access all the genetic resources in the triangle of U for genetic improvement of existing species and development of new agricultural species. Each of the three Brassica genomes occurs in several species, where they are distinguished as subgenomes with a tag to identify the species of origin. For example, the A subgenome in B. juncea (2n = AABB) is denoted as A^j and the A subgenome in B. napus (2n = AACC) as Aⁿ. Trigenomic bridges have been used to increase genetic diversity in allopolyploid Brassica crop species, such as a new-type B. napus with subgenomes from B. rapa (A^r) and B. carinata (C^c). Recently, trigenomic bridges from several sources have been crossed together as the ‘founders’ of a potentially new allohexaploid Brassica species (AABBCC). During meiosis in a trigenomic bridge, crossovers are expected to form between homologous chromosomes of related subgenomes (for example A^r and Aⁿ), but cross-overs may also occur between non-homologous chromosomes (for example between A and C genome chromosomes). Irregular meiosis is a common feature of new polyploids, and any new allotetraploid or allohexaploid Brassica genotypes derived from a trigenomic bridge must achieve meiotic stability through a process of diploidisation. New sequencing technologies, at the genomic and epigenomic level, may reveal the genetic and molecular basis of diploidization, and accelerate selection of stable allotetraploids or allohexaploids. Armed with new genetic resources from trigenomic bridges, Brassica breeders will be able to improve yield and broaden adaptation of Brassica crops to meet human demands for food and biofuel, particularly in the face of abiotic constraints caused by climate change.     

Somatic hybrids between Brassica juncea (L). Czern. and Diplotaxis harra (Forsk.) Boiss and the generation of backcross progenies

An attempt to transfer genes from droughttolerant Diplotaxis harra, a wild relative of Brassica species, to an elite oil-yielding cultivar, B-85, of mustard (Brassica juncea) was made through protoplast fusion, as the two plant systems are sexually incompatible. By following the standard protocol for PEG-mediated protoplast fusion followed by high pH, high Ca⁺⁺, DMSO treatment and appropriate cell-culture technique, 16 presumptive somatic hybrid plants could be regenerated. Chromosomal analysis of four such somatic hybrids revealed that three of them were asymmetric. Analysis of morphological characters, meiotic chromosomes, and esterase isoenzyme pattern revealed that all the somatic hybrids were different from each other. Furthermore four chromosomes of each genome could undergo homoeologous pairing at meiosis indicating the possibilities for genetic recombination and chromosomal rearrangements. Irregular distribution of chromosomes at anaphase-II at meiosis has been a consistent feature of these plants. Eventually, pollen of all the somatic hybrids showed complete infertility preventing the recovery of any selfed seed. Nevertheless, ovule fertility of one somatic hybrid was not totally impaired as it had set some seeds upon backcrossing with the B. juncea parent. The esterase isoenzyme banding pattern of 24 individual progeny plants of this backcross provided evidence for their recombinant nature. It was thus confirmed that a transfer of genetic traits from Diplotaxis harra to B. juncea had indeed taken place. Furthermore, it was conceptualised that a transfer of alien genes through the protoplast-fusion technique is primarily possible in situations where meiotic pairing of the chromosomes of the two participating genomes generates recombinant gametocytes which can pass through subsequent filial generations.     

Production and cytogenetics of intergeneric hybrids between the three cultivated Brassica diploids and Orychophragmusviolaceus

It has been proposed that both complete and partial separation of the parental genomes during mitosis and meiosis occurs in the intergeneric hybrids between Orychophragmus violaceus (2n=24) and the three cultivated Brassica tetraploids (B. napus, B. carinata and B. juncea). The hypothesis has been that this and the variations in chromosome numbers of these hybrids and their progenies result from the different roles of the A, B and C genomes originating from Brassica. To test this hypothesis, we produced hybrids between O. violaceus and the cultivated Brassica diploids. The hybrids with B. oleracea (2n=18, CC) had an intermediate morphology, but their petals were purple like those of O. violaceus. They were sterile and had the expected chromosome number (2n=21) in their mitotic and meiotic cells. The hybrid with B. campestris (2n=20, AA) was morphologically intermediate, except for its partial fertility and its yellow petals, which were similar to those of B. campestris. It was mixoploid (2n=23–42), and cells with 2n=34 were most frequent. Partial separation of parental genomes during mitosis, leading to the addition of O. violaceus chromosomes to the B. campestris complement, was proposed to explain the findings in the mitotic and meiotic cells of the hybrid and its progeny. In crosses with B. nigra (2n=16, BB), the majority of the F₁ plants were of the maternal type (2n=16), a small fraction had B. nigra morphology but were mixoploids (2n=16–18), predominantly with 2n=16 cells and three plants, each with a specific morphology, were mixoploids consisting of cells with varying ranges of chromosome numbers (2n=17–26, 11–17 and 14–17). The origin of these different types of plants was inferred to be a result of the complete and partial separation of parental genomes and the loss of O. violaceus chromosomes. Our findings in the three crosses suggest that the A genome was more influential than the C genome with respect to complete genome separation during mitosis and meiosis of the hybrids with B. napus. Possible complete and partial genome separation during mitotic divisions of the hybrids with B. carinata was mainly attributed to the role of the B genome. The combined roles of the A and B genomes would thus contribute to the most variable chromosome numbers of mitotic and meiotic cells in the hybrids with B. juncea and their progenies. The possible cytological mechanisms pertaining to these hybrids and the potential of genome separation in the production of Brassica aneuploids and homozygous plants are discussed. Received: 8 February 1998 / Accepted: 12 March 1999     

Broadening the avenue of intersubgenomic heterosis in oilseed Brassica

Accumulated evidence has shown that each of the three basic Brassica genomes (A, B and C) has undergone profound changes in different species, and has led to the concept of the “subgenome”. Significant intersubgenomic heterosis was observed in hybrids between traditional Brassica napus and first generation lines of new type B. napus. The latter were produced by the partial introgression of subgenomic components from different species into B. napus. To increase the proportion of exotic subgenomic components and thus achieve stronger heterosis, lines of first generation new type B. napus were intercrossed with each other, and subjected to intensive marker-assisted selection to develop the second generation of new type B. napus. The second generation showed better agronomic traits and a higher proportion of introgression of subgenomic components than did the first generation. Compared with the commercial hybrid and the hybrids produced with the first generation new type B. napus, the novel hybrids showed stronger heterosis for seed yield during the 2 years of field trials. The extent of heterosis showed a significant positive correlation with the introgressed subgenomic components in the parental new type B. napus. To increase the content of the exotic subgenomic components further and to allow sustainable breeding of novel lines of new type B. napus, we initiated the development of a gene pool for new type B. napus that contained a substantial amount of genetic variation in the A^r and C^c genome. We discuss new approaches to broaden the avenue of intersubgenomic heterosis in oilseed Brassica.     

Transfer of Brassica tournefartii (TT) genes to allotetraploid oilseed Brassica species (B. juncea AABB, B. napus AACC, B. carinata BBCC): homoeologous pairing is more pronounced in the three-genome hybrids (TACC, TBAA, TCAA, TCBB) as compared to allodiploids (TA, TB, TC)

For the transfer of genes from B. tournefortii (TT) to the allotetraploid oilseed brassicas, B. juncea AABB, B. carinata BBCC and B. napus AACC, B. tournefortii was first crossed with the three basic diploid species, B. campestris (AA), B. nigra (BE) and B. oleracea (CC), to produce the allodiploids TA, TB and TC. These were tetraploidized by colchicine treatment to produce the allotetraploids TTAA, TTBB and TTCC, which were further crossed with B. juncea and B. napus to produce three-genome hybrids with substitution-type genomic configurations: TACC, TBAA and TCAA. These hybrids along with another hybrid TCBB produced earlier, the three allodiploids, their allotetraploids and the four diploid parent species were studied for their male meiotic behaviour. The diploid parent and the allotetraploids (TTAA, TTBB and TTCC) showed regular meiosis although the pollen viability was generally low in the allotetraploids. In the allodiploids (TA, TB and TC) only some end-to-end associations were observed without any clearly discernible chiasmata or exchange points. Chromosomes involved in end-to-end associations were randomly distributed at the metaphase/anaphase-I stages. In contrast, the three-genome hybrids (TACC, TBAA, TCAA and TCBB) showed normal bivalents whose number exceeded the expected bivalent values. Bivalents arising out of homoeologous pairing were indistinguishable from normal pairs by their disjunction pattern but could be distinguished on the basis of the heteromorphy of the homoeologous chromosomes. The three-genome hybrids could be backcrossed to allotetraploid oilseed brassicas as they had some fertility. In contrast, the allodiploids could neither be selfed nor back-crossed. On the basis of their meiotic stability, in terms of more pronounced homoeologous pairing and fertility for backcrossing, the three-genome configurations provide the best possible situation for the introgression of alien genes from the secondary gene pool to the allotetraploid oilseed crops B. juncea, B. napus and B. carinata.     

A novel intergeneric hybrid in the Triticeae: Triticum aestivum x Psathyrost achys juncea

Summary Two hybrid embryos of intergeneric origin between Triticum aestivum cv Fukuho (2n=6x=42, AABBDD) and Psathyrostachys juncea (2n=2x=14, NN) were successfully rescued. One hybrid plant had the expected chromosome number of 28 (ABDN), whereas the second plant had 35 chromosomes. The average meiotic chromosome pairing in the 35-chromosome hybrid was 21.87 univalents + 6.38 bivalents + 0.11 trivalents + 0.009 quadrivalents, which indicates that two copies of the N genome were present. Chromosome pairing in the 28-chromosome hybrid was low (1.35 bivalents), and pointed out the lack of homology between the wheat genomes and the P. juncea genome. These new hybrids showed some necrosis and chlorosis, which caused severe floral abortion in the plant that had 35 chromosomes. These problems became gradually less severe after 18 months.Contrib. no. 372     

SYNTHETIC HYBRIDS OF NEW WORLD AND OLD WORLD AGROPYRONS. IV. TETRAPLOID AGROPYRON SPICATUM F. INERME X TETRAPLOID AGROPYRON DESERTORUM

Emasculated and unemasculated crosses of tetraploid A. spicatum f. inerme X A. desertorum yielded four hybrids. The hybrids were morphologically intermediate between the parent species but resembled A. desertorum more closely than A. spicatum. Both parents behaved cytologically as autoploids. Mean chromosome associations of 0.04 I, 8.60 II, 0.01 III, and 2.67 IV were observed at diakinesis in the 28-chromosome A. spicatum. The A. desertorum parent contained 30 chromosomes, 2 of which were likely supernumeraries, and averaged 0.03 I, 9.85 II, and 2.57 IV at diakinesis. Three hybrids contained 30 chromosomes, and one had 29. The most common chromosome association in the 30-chromosome hybrids was 2 I and 14 II; and the average was 3.00 I, 13.40 II, 0.06 III, and 0.01 IV. A. spicatum and A. desertorum chromosomes were usually distinguishable from each other in the hybrid cell on the basis of size. All pairing in the hybrids was attributed to autosyndesis within parental genomes. A. spicatum, A. desertorum, and their hybrids were represented by genome formulas of SSSS, CCCC, and SSCC, respectively. The hybrids produced 5 to 439 seeds under open pollination. Three controlled crosses between the hybrids yielded 2, 5, and 23 seeds, respectively, on 10 maternal spikes in each cross. The prospects of developing a fertile, cytologically stable allotetraploid species from the hybrids appear favorable.     

Unique chromosome behavior and genetic control in Brassica×Orychophragmus wide hybrids: a review

Researchers recognized early that chromosome behavior, as other morphological characters, is under genetic control and gave some cytogenetical examples such as the homoeologous chromosome pairing in wheat. In the intergeneric sexual hybrids between cultivated Brassica species and another crucifer Orychophragmus violaceus, the phenomenon of parental genome separation was found under genetic control during mitosis and meiosis. The cytogenetics of these hybrids was species-specific for Brassica parents. The different chromosome behavior of hybrids with three Brassica diploids (B. rapa, B. nigra and B. oleracea) might contribute to the different cytology of hybrids with three tetraploids (B. napus, B. juncea and B. carinata). The finding that genome-specific retention or loss of chromosomes in hybrids of O. violaceus with B. carinata and synthetic Brassica hexaploids (2n=54, AABBCC) is likely related to nucleolar dominance gives new insight into the molecular mechanisms regarding the cytology in these hybrids. It is proposed that the preferential expressions of genes for centromeric proteins from one parent (such as the well presented centromeric histone H3) are related with chromosome stability in wide hybrids and nucleolar dominance is beneficial to the production of centromere-specific proteins of the rRNAs-donor parent and to the stability of its chromosomes.     

Tumbleweed (Salsola, section Kali) species and speciation in California

Tumbleweeds (Salsola species, section Kali) are road side and rangeland pest plants throughout the 48 contiguous states in the US. Three described tumbleweed species and two undescribed Salsola taxa occur in California. The known species are Russian thistle, Salsola tragus, introduced from Eurasia in the 1800s, Russian barbwire thistle, S. paulsenii, which grows in the desert regions of California, and is also native to Eurasia, and the recently identified S. kali subspecies austroafricana, possibly native to South Africa. Our goals were to investigate karyology, genome size, and molecular genetic affinities of the described species and the other taxa within their ranges in California using recently developed microsatellite loci, dominant nuclear DNA markers (RAPD and ISSR), and DNA sequence data. Chromosome counts and genome size assessments made with flow cytometry were compared. These analyses indicated that one undescribed taxon is a new allopolyploid hybrid between S. tragus and S. kali subspecies austroafricana, and the other undescribed taxon appears to be a complex hybrid involving all three described species. The invasion potentials for the hybrid taxa are unknown. Tumbleweeds are the focus of biological controls efforts but the identification of suitable agents for the hybrid taxa may be problematic because of the large amount of genetic variability encompassed within this evolving Salsola complex.     

Seed glucosinolates of fourteen wild Brassica species

Seed glucosinolates have been determined for 14 wild Brassica species, by micro-scale GC analysis of silylated derivatives. Of these, 12 were investigated for the first time. The majority of taxa exhibited high alkenylglucosinolate levels, although prop-2-enylglucosinolate appears to be generally absent. Other known methionine-derived glucosinolates predominate in B. tournefortii, B. elongata and B. deflexa. Phenylalanine-derived 4-hydroxybenzylglucosinolate is characteristic of section Brassicaria plants and represents the first finding of this glucosinolate in authenticated Brassica material.     

A Brassica campestris-alboglabra addition line and its use for gene mapping,intergenomic gene transfer and generation of trisomics

Summary Brassica campestris-alboglabra monosomic addition lines were developed from a trigenomic Brassica hybrid (2 n=3 x=29, AAC) obtained by backcrossing a resynthesized B. napus (2 n=4 x=38, AACC) line to its parental B. campestris (2 n=2 x=20, AA) line. One addition line was characterized genetically with three loci specific for the alien chromosome and cytologically by meiotic analysis. The following results were obtained. (1) The same chromosome in the B. alboglabra (2 n= 2 x=18, CC) genome carried the three loci, E _c, W _c and Lap-1 C ^c, which control the biosynthesis of erucic acid, white flower colour and the faster migrating band of leucine aminopeptidase, respectively. The linear order and possible positions of the three loci were inferred. The meiotic behaviour of the alien chromosome was documented and its transmission frequency was assessed. (2) Intergenomic recombination frequently occurred in the monosomic addition line, resulting in the introgression of one or two loci from the alien chromosome into the B. campestris genome. (3) B. campestris trisomics were found in the progeny of the monosomic addition line. (4) The removal of the other eight C-genome chromosomes from the trigenomic Brassica hybrid led to a dramatic increase in the erucic acid content of the monosomic addition line. (5) No offspring of the trigenomic Brassica hybrid showed evidence of intergenomic recombination and introgression of the W _c locus into the B. campestris genome. It is questioned whether such a difference might be due to a possible regulating mechanism for homoeologous chromosome pairing.     

Somatic hybrids with substitution type genomic configuration TCBB for the transfer of nuclear and organelle genes from Brassica tournefortii TT to allotetraploid oilseed crop B. carinata BBCC

Oilseed crop Brassica carinata BBCC is a natural allotetraploid of diploid species B. nigra BB and B. oleracea CC. To transfer the nuclear and organelle genes in a concerted manner from an alien species, B. tournefortii TT, to B. carinata, we produced somatic hybrids with genomic configuration TCBB using B. nigra and B. oleracea stocks that carried selectable marker genes. B. tournefortii TT was sexually crossed with hygromycin-resistant B. oleracea CC. Protoplasts isolated from shoot cultures of hygromycin-resistant F₁ hybrids of B. tournefortiixB. oleracea TC were fused with protoplasts of kanamycin-resistant B. nigra BB. In two different fusion experiments 80 colonies were obtained through selection on media containing both hygromycin and kanamycin. Of these, 39 colonies regenerated into plants. Analysis of 15 regenerants by random amplified polymorphic DNA (RAPD) markers showed the presence of all three genomes, thereby confirming these to be true hybrids. Restriction fragment length polymorphism (RFLP) analysis of organelle genomes with heterologous chloroplast (cp)and mitochondrial (mt) DNA probes showed that the chloroplast genome was inherited from either of the two parents while mitochondrial genomes predominantly showed novel configurations due to either rearrangements or intergenomic recombinations. We anticipate that the TCBB genomic configuration will provide a more conducive situation for recombination between the T and C genomes during meiosis than the TTCCBB or TCCBB type configurations that are usually produced for alien gene transfer. The agronomic aim of producing TCBB hybrids is to transfer mitochondrial genes conferring cytoplasmic male sterility and nuclear genes for fertility restoration from B. tournefortii to B. carinata.     

Crossovers Get a Boost in Brassica Allotriploid and Allotetraploid Hybrids

Meiotic crossovers are necessary to generate balanced gametes and to increase genetic diversity. Even if crossover number is usually constrained, recent results suggest that manipulating karyotype composition could be a new way to increase crossover frequency in plants. In this study, we explored this hypothesis by analyzing the extent of crossover variation in a set of related diploid AA, allotriploid AAC, and allotetraploid AACC Brassica hybrids. We first used cytogenetic methods to describe the meiotic behavior of the different hybrids. We then combined a cytogenetic estimation of class I crossovers in the entire genome by immunolocalization of a key protein, MutL Homolog1, which forms distinct foci on meiotic chromosomes, with genetic analyses to specifically compare crossover rates between one pair of chromosomes in the different hybrids. Our results showed that the number of crossovers in the allotriploid AAC hybrid was higher than in the diploid AA hybrid. Accordingly, the allotetraploid AACC hybrid showed an intermediate behavior. We demonstrated that this increase was related to hybrid karyotype composition (diploid versus allotriploid versus allotetraploid) and that interference was maintained in the AAC hybrids. These results could provide another efficient way to manipulate recombination in traditional breeding and genetic studies.     

Interspecific hybridization inBrassica: Application of flow cytometry for analysis of ploidy and genome composition in hybrid plants

Interspecific hybrids from the crosses betweenBrassica campestris, B. carinata, B. juncea andB. napus were obtained throughin vitro ovary and ovule culture. F₁ hybrids were studied morphologically and flow cytometry was used to estimate 2C nuclear DNA content both in parentalBrassica species and their hybrids. It was found that in comparison with the A genome, the B and the C genomes ofBrassica contained 26.9 % and 43.9 % more DNA, respectively. This finding may be used to distinguish interspecific hybrids containing various genome combinations. It was concluded that flow cytometric analysis of nuclear DNA content might be useful tool inBrassica breeding.     

Intergeneric (intersubtribe) hybridization between Moricandia arvensis and Brassica A and B genome species by ovary culture

Summary Intergeneric hybrids between Moricandia arvensis (C₃–C₄ intermediate species) and Brassica A and B genome species (B. campestris and B. nigra) were produced via ovary culture. When M. arvensis was used as a female parent, the hybrid embryo yield (0.25–0.45 embryo per pollination) was similar between two genomes, regardless of the male parent. The reciprocal hybrid using B. campestris as a female was also obtained, although yield of embryo was lower (0.02 embryo per pollination). On the other hand, no hybrids were obtained without the in vitro technique. As most hybrid embryos could not develop normal shoots, plants were regenerated by inducing shoots on the cultured hypocotyl. The hybrid nature of the regenerated plant was confirmed morphologically and cytogenetically. A certain amount of bivalents (2.52-2.71) in the hybrids indicated the existence of partial chromosome homology between two genera. The present results indicate that ovary culture is an effective technique for overcoming the crossing barrier between M. arvensis and Brassica cultivated species.     

Hybrids and backcross progenies between wheat (Triticum aestivum L.) and apomictic Australian wheatgrass [Elymus rectisetus (Nees in Lehm.) A. Löve & Connor]: karyotypic and genomic analyses

Wheat (Triticum aestivum L., 2n = 6x = 42, AABBDD) florets were emasculated and pollinated using two apomictic wheatgrass [Elymus rectisetus (Nees in Lehm.) A. Love & Connor, 2n = 6x = 42, SSYYWW] accessions, one of which produces 2n pollen. A 2n = 42 (B_II) hybrid and four 2n = 63 (B _III) hybrids were obtained. The spike morphology of the B _II hybrid was intermediate to that of its parents. The pollen mother cells (PMCs) of this hybrid contained on average 38.361 and 1.62 II, which was consistent with its disparate genome composition (ABDSYW). Its pollen failed to stain and no BC₁ progeny was obtained. The B _III hybrids (reduced egg fertilized with unreduced sperm) were grasslike and had a full complement of E. rectisetus chromosomes, the synapsis of which was slightly impaired by wheat haplome and/or cytoplasm. Their PMCs contained on average 16.30 II, 25.72 I, and 1.54 multivalents (III plus IV). Pollen stainability in these hybrids was low (<1%), and when they were used as females, one 54- and 60-chromosome BC₁ were obtained. A mean of 13.25 II was observed in PMCs of the 54-chromosome BC₁ and pollen stainability was 10%. Pollen stainability in the 60-chromosome BC₁ was only 5%. The use of 2n-pollen-producing E. rectisetus accession accelerated hybrid and BC₁ formation and may accelerate the ultimate transfer of apomixis to wheat.