Production and characterization of intergeneric hybrids between Brassica oleracea and a wild relative Moricandia arvensis

Intergeneric crosses were made between Brassica oleracea and Moricandia arvensis utilizing embryo rescue. Six F₁ hybrid plants were generated in the cross‐combination of B. oleracea × M. arvensis from 64 pods by the placenta‐embryo culture technique, whereas three plants were produced in the reciprocal cross from 40 pods by the ovary culture technique. The hybrid plants were ascertained to be amphihaploid with 2n = 23 chromosomes in mitosis and a meiotic chromosome association of (0–3)II + (17–23)I at metaphase I (M I). In the backcross with B. oleracea, some of these hybrids developed sesquidiploid BC₁ plants with 2n = 32 chromosomes that predominantly exhibited a meiotic configuration of (9II + 14I) in pollen mother cells. The following backcross of BC₂ plants to B. oleracea generated 48 BC₃ progeny with somatic chromosomes from 2n = 19 to 2n = 41. The 2n = 19 plants showed a chromosomal association type of (9II + 1I) and a chromosomal distribution type of (9¹/₂ + 9¹/₂) or (9 + 10) at M I and M II, respectively. These facts might suggest that they were monosomic addition lines (MALs) of B. oleracea carrying a single chromosome of M. arvensis that could offer potential for future genetic and breeding research, together with other novel hybrid progeny developed in this intergeneric hybridization.     

单体附加系甜菜及空中搭载实验材料相关性状的研究

为了探明远缘杂交甜菜单体附加系M14高频传递的遗传学机理,通过胚珠分离切割、花粉败育调查和含糖量单株检测等方法,对单体附加系甜菜及空中搭载实验材料的性状进行了分析研究。研究结果验证了单体附加系甜菜M14高频传递的遗传特性存在兼性无融合生殖现象。无融合生殖体的比例达96.5%,花粉败育程度高达90%以上,大孢子母细胞通过有丝分裂形成胚囊不需要精卵结合繁育后代;有性生殖体的比例占3.5%, 有性生殖体存在致死基因的可能性。单体附加系甜菜及空中搭载的实验样品含糖量均高于普通栽培甜菜。野生白花甜菜染色体携带的高糖、抗病、无融合生殖等基因决定了远缘杂交甜菜和空中搭载实验材料具有专属的遗传学性状。研究结果为进一步开拓野生白花甜菜有益基因进行染色体工程研究奠定了良好的基础。     

Moricandia arvensis cytoplasm based system of cytoplasmic male sterility in Brassica juncea: Reappraisal of fertility restoration and agronomic potential

Cytoplasmic male sterility (CMS) system based on the cytoplasm from Moricandia arvensis (mori) was investigated for fertility restoration and agronomic potential. Fertility restorer gene for mori CMS was introgressed from cytoplasm donor species as all the evaluated Brassica juncea genotypes (155) acted as sterility maintainers. The allosyndetic pairing between M^a and the A/B genome chromosomes in the monosomic addition plants (2n= 18II + 1M^a) facilitated the gene introgression. Partial fertility restoration (43–52% pollen grain stainability) in F₁ hybrids and absence of segregation for male sterility in F₂ progenies suggested gametophytic control of fertility restoration. The pollen fertility in the F₁ hybrids was, however, sufficient to ensure complete seed set upon bag selfing. Introgression from M. arvensis also helped in correction of chlorosis associated with mori cytoplasm in CMS and fertile alloplasmic B. juncea plants. Yield evaluation of thirty F₁ hybrids having the same nuclear genotype but varied male sterilizing cytoplasms (mori, oxy, lyr, refined ogu), in comparison to respective euplasmic hand bred control hybrids, allowed an estimate of yield penalty associated with different CMS systems. It ranged from 1.8% to 61.6%. Hybrids based on cytoplasmically refined ogu were most productive followed by those based on cytoplasmically refined mori CMS. The male sterility systems emanating from somatic hybridization were found superior than those developed from sexual hybridization.     

Production and characterization of an alloplasmic and monosomic addition line of Brassica rapa carrying the cytoplasm and one chromosome of Moricandia arvensis

Intergeneric hybridization was performed between Moricandia arvensis and four inbred lines of Brassica rapa following embryo rescue. Three F₁ hybrid plants were developed from three cross combinations of M. arvensis × B. rapa, and amphidiploids were synthesized by colchicine treatment. Six BC₁ plants were generated from a single cross combination of amphidipolid × B. rapa ‘Ko1-303’ through embryo rescue. One BC₂ and three BC₃ plants were obtained from successive backcrossing with B. rapa ‘Ko1-303’ employing embryo rescue. Alloplasmic and monosomic addition lines of B. rapa (Allo-MALs, 2n = 21) were obtained from backcrossed progeny of three BC₃ plants (2n = 21, 22 and 23) without embryo rescue. An alloplasmic line of B. rapa (2n = 20) degenerated before floliation on 1/2 MS medium due to severe chlorosis. Allo-MALs of B. rapa (2n = 21) showed stable male sterility without any abnormal traits in vegetative growth and female fertility. Molecular analyses revealed that the same chromosome and cytoplasm of M. arvensis had been added to each Allo-MAL of B. rapa. This Allo-MAL of B. rapa may be useful material for producing cytoplasmic male sterile lines of B. rapa.     

Sexual hybridization between Capsella bursa‐pastoris (L.) Medik (♀) and Camelina sativa (L.) Crantz (♂) (Brassicaceae)

The development of transgenic oilseed Camelina sativa (2n = 40) and the potential for hybridization with its weedy relative Capsella bursa‐pastoris (2n = 36) necessitates a careful evaluation of the reproductive compatibility between the species. Here, we conducted over 1800 crosses (emasculation and manual pollination) to examine the ability of 10 Canadian C. bursa‐pastoris (♀) accessions to hybridize with five accessions of C. sativa (♂). Seven hybrids were confirmed among 586 putative hybrids screened with species‐specific markers, indicating a hybridization rate of 1.5 hybrids per 10 000 ovules pollinated. All seven hybrids had intermediate DNA content compared to their parents, were morphologically distinct, had low (1.9%) pollen fertility and failed to produce selfed or backcrossed seed. Given the abundance of C. bursa‐pastoris along field margins, hybrids will likely be generated in the wild, but they will be unable to establish lineages unless fertility is restored. The large number of crosses and the diversity captured by the use of multiple accessions resulted in strong statistical power and a high degree of confidence in the estimated hybridization rate.     

Incongruity of interspecific and intergeneric crosses involving Nicotiana and Petunia species that exhibit potential for somatic hybridization

Summary Extensive pollinations were made in attempts to produce selected intergeneric or interspecific hybrids using Nicotiana and Petunia species that exhibit potential for somatic hybridization. Pollination between pairs of species was done by standard and bud-pollination methods. Nicotiana alata or N. tabacum when pollinated reciprocally with selected Petunia species failed to produce hybrids. Likewise, no interspecific hybrids were obtained between Petunia parviflora and 4 Petunia species or P. hybrida. The type of incongruity existing between these species is discussed in relation to the production of hybrids by standard In vitro techniques as compared to somatic hybridization.On leave from the Department of Horticulture, Michigan State University, E. Lansing, Michigan, USA. supported by a European Molecular Biology Organization (EMBO) fellowship and The Fred C. Gloeckner Foundation, N. Y., USA.Supported by the Agricultural Research Council.     

Morphological and molecular analysis of androgenetic, selfed and backcrossed plants produced from a Hordeum vulgare x H, bulbosum hybrid

Anther culture (AC) was carried out on a fertile triploid hybrid between Hordeum vulgare L. (cultivated barley) and H. bulbosum L, (bulbous barley grass) to determine whether AC-derived regenerants differed from progeny obtained through selfing and backcrossing. Chromosome counts were carried out on all plants and DNA was extracted from them to prepare Southern blots for molecular analysis. To identify true recombinants, the blots were probed with rye repetitive sequence probes (pSc119.1 and pScl19.2). which hybridize strongly and specifically to H. bulbosum DNA. Twenty probes that detect single- or low-copy sequences were hybridized with Southern blots containing restricted DNA extracted from 25 AC-derived plants, 11 selfed and six backcrossed progeny that showed hybridizations with pScll9. Although restriction fragment length polymorphisms (RFLPs) were only observed using probes that map to four of the possible 14 chromosome arms, an introgression associated with chromosome 6HS was frequently observed among plants derived from AC. selfing and backcrossing. Plants from AC differed from selfed and backcrossed progeny in their chromosome number; unique RFLP bands that were occasionally observed may indicate chromosomal rearrangements.     

Interspecific hybridization of a multiploid mutant of durum wheat with rye and Triticum monococcum L. results in pentaploid hybrids

The multiploid mutant of durum wheat is a genotype that produces unreduced gametes. Our objective was to test the recovery of pentaploid hybrids in crosses of the mutant with rye and Triticum monococcum L. Compared with check crosses, the mutant had a two‐third reduction in percent seed set for rye crosses, but had only a slight decrease in crossability with T. monococcum. Pentaploid hybrids were associated with plump seeds of the mutant/rye cross, and with shrivelled seeds of the mutant/T. monococcum cross. We suggest that the endosperm balance number hypothesis explains the association of pentaploid hybrids with endosperm type. This association made for easy recovery of pentaploid hybrids from crosses to both species. Mature, plump seeds from the mutant/rye cross were germinated and pentaploid hybrids were recovered. One pentaploid hybrid was recovered for every 50.5 and 15.1 florets pollinated with rye and T. monococcum, respectively. Unreduced gametes in the multiploid mutant will facilitate interspecific hybridization by reducing the time to produce pentaploid plants.     

Congruity backcrossing as a means of creating genetic variability in self pollinated crops: seed morphology of Phaseolus vulgaris L. and P. acutifolius A. Gray hybrids

Summary Seeds of early generations of three reciprocal congruity-backcross (CBC) pedigrees, developed by backcrossing Phaseolus vulgaris-P. acutifolius hybrids to each of the parent species in alternate generations, exhibited a preponderance of traits (size, shape, color, and pattern) of the cytoplasmic parent. The large size of Red Cloud (V₁), the P. vulgaris parent common to all of the pedigrees, dominated pedigrees with V₁ as the cytoplasmic parent, while the small size and rounded or square shapes of the tepary parents, wild P. acutifolius var. acutifolius PI 263590 or G400445 (A₁₉), wild P. acutifolius var. latifolius PI 406622 (A₁₀), or cultivated P. acutifolius var. latifolius Serowi PI 319443 (A₉), were the majority phenotypes when P. acutifolius was the cytoplasmic parent. Continuing through the second cycle of CBC, that is the second backcross with each of the parent species or the fourth backcross, began an amelioration of the apparent cytoplasmic effect on gene expression, as reciprocal pedigrees became more alike, usually with intermediate expression of parental traits or the appearance of new traits. The large seed size of V₁ was recovered in hybrids with P. acutifolius cytoplasm and the kidney shape of V₁ became rare in hybrids with P. vulgaris cytoplasm. Although the tepary-bean parents represented two subspecies and both cultivated and wild P. acutifolius, the three sets of reciprocal-hybrid pedigrees with P. vulgaris Red Cloud are surprisingly similar. It may be that the exotic parent used to develop a CBC pedigree should be selected more for combining ability in the interspecific cross than for specific economic traits. While the number of generations (six or more) required to produce fertile, intermediate CBC hybrids (that did not require embryo rescue) may preclude routine use of this method by practical plant breeders, the crossability of advanced hybrids with both parental species and the amount of variability apparent in advanced-hybrids progenies suggests that CBC would be valuable for maintaining exotic germplasm in immediately useful forms.     

Field assessment of a second generation backcross (BC1 × Passiflora edulis) of passion fruit for agronomic performance and resistance to CABMV

The objective of this work was to characterize and identify among the passion fruit hybrids of the second generation backcross (BC2: BC1 × P. edulis), resistance to passion fruit woodiness disease (CABMV) with desirable agronomic attributes. Ninety‐two progeny of BC2 hybrids were evaluated for severity of the virus in conditions of field, physical and chemical characterization of the fruits and precocity of the plant in relation to the vegetative and reproductive phases. The data were analysed using multivariate statistics and mixed models (REML/BLUP) to estimate genetic parameters. The was conditioned by and , for number of fruits, fruit and peel weight and soluble solids and the effect of for virus severity to CABMV and effect of on the variables fruit length, fruit diameter, peel thickness and pulp weight. For the titratable acidity it had no . The progeny BC2‐A, BC2‐C, BC2‐E, BC2‐F and BC2‐H families can be selected for use in passion fruit breeding programmes by adding high vegetative vigour, early fruit production, resistance to CABMV and also desirable agronomic attributes.     

Simple Genetic Control of Hybrid Plant Development in Interspecific Crosses between Phaseolus vulgaris L. and P. acutifolius A. Gray

Crosses were performed between nine Phaseolus vulgaris lines (as females) and seven P. acutifolius lines (as- male to examine parental compatibility for the production of vigorous hybrid And backcross plants, in vitro embryo rescue techniques were required to secure hybrid and backcross proseny following interspecific crossing. Seedling development appeared to be dependent on which allele the P. vulgaris parent carried at an interspecific incompatibility locus. Seven of the nine P. vulgaris lines tested carried an allele at this locus which interacted with a nuclear factor in the P. acutifolius genome resulting in stunted, sub-lethal hybrids. The lines, ICA pijao' and ‘Sacramento Light Red Kidney’ did not carry this allele and produced vigorous hybrid progeny in combination with all P. acutifolius parents. Intensive backcrossing produced progeny which also segregated for sub-lethal and viable plant development. The observed segregation patterns suggest that a bridge crossing scheme would facilitate the introgression of P. acutifolius germplasm into incompatible P. vulgaris lines. Similarities, with an intraspecific incompatibility system are discussed.     

Interspecific hybridization of Brassica napus and B. juncea through ovary,ovule and embryo culture

Summary Employing in vitro culture of ovaries, ovules and embryos, interspecific hybrids have been obtained amongst two important oilseed crops, Brassica napus x B. juncea and their reciprocal. The test-tube hybrid plants have been transferred to the field, and reared to maturity. The F₁ seeds obtained from the hybrid ovaries showed normal germination, and the hybrid plants exhibited a range of variation of characters.     

Chromosome and chromosomal arm locations of genes for resistance to Septoria glume blotch in wheat cultivar Cotipora

Summary Septoria glume blotch, caused by Stagonospora nodorum, is an important disease of wheat (Triticum aestivum). Separate genetic mechanisms were found to control flag leaf and spike resistance. Genes for resistance to S. nodorum were located on different chromosomes in the few wheat cultivars studied. These studies only partially agree on the chromosome locations of gene in wheat for resistance to S. nodorum, and chromosomal arm locations of such genes are not known. The objectives of this study were to determine the chromosome and chromosomal arm locations of genes that significantly influence resistance to S. nodorum in wheat cultivar Cotipora. Monosomic analysis showed that flag leaf resistance was controlled by genes on chromosomes 3A, 4A, and 3B whereas the spike resistance was controlled by genes on chromosomes 3A, 4A, 7A, and 3B (P=0.01). Additionally, genes on chromosomes 6B and 5A influenced the susceptibility of the flag leaf and spike reactions, respectively (P=0.01). Telocentric analysis showed that genes on both arms of chromosome 3A, and the long arms of chromosomes 4A and 3B were involved in the flag leaf resistance whereas genes on both arms of chromosome 4A, the short arm of chromosome 3A, and the long arm of chromosome 3B conferred spike resistance.     

Intergeneric hybridization of Diplotaxis erucoides with Brassica napus. I. cytogenetic analysis of F1 and BC1 progeny

Summary Intergeneric hybrids (F₁) Diplotaxis erucoides (DeDe) x Brassica napus (AACC) and the first backcross to B. napus (BC₁) have been obtained through in vitro culture of excised ovaries. The chromosome numbers of F₁ and BC₁ plants proved the occurrence of unreduced gametes. The study of metaphase I chromosome pairing showed that autosyndesis in De genome and allosyndesis between De and A/C genomes might exist. The male fertility of the F₁ plants was low. Some male-sterile plants were found in F₁ and BC₁ progeny. The possibilities of creating addition lines B. napus-D. erucoides and of obtaining a new cytoplasmic male sterility in B. napus are discussed.     

Cytological analyses of hybrids and derivatives of hybrids between durum wheat and Thinopyrum bessarabicum,using multicolour fluorescent GISH*

The wheat progenitors and other wild relatives continue to be important sources of genes for agronomically desirable traits, which can be transferred into durum wheat (Triticum turgidum; 2n = 4x = 28; AABB genomes) cultivars via hybridization. Chromosome pairing in durum × alien species hybrids provides an understanding of genomic relationships, which is useful in planning alien gene introgression strategies. Two durum cultivars, ‘Lloyd’ and ‘Langdon’, were crossed with diploid wheatgrass, Thinopyrum bessarabicum (2n = 2x = 14; JJ), to synthesize F₁ hybrids (2n = 3x = 21; ABJ) with Ph1. ‘Langdon’ disomic substitution 5D(5B) was used as a female parent to produce F₁ hybrids without Ph1, which resulted in elevation of pairing between durum and grass chromosomes – an important feature from the breeding standpoint. The F₁ hybrids were backcrossed to respective parental cultivars and BC₁ progenies were raised. ‘Langdon’ 5D(5B) substitution × Th. bessarabicum F₁ hybrids were crossed with normal ‘Langdon’ to obtain BC₁ progeny. Chromosome pairing relationships were studied in F₁ hybrids and BC₁ progenies using both conventional staining and fluorescent genomic in situ hybridization (fl‐GISH) techniques. Multicolour fl‐GISH was standardized for characterizing the nature and specificity of chromosome pairing: A–B, A–J and B–J pairing. The A–J and B–J pairing will facilitate gene introgression in durum wheat. Multicolour fl‐GISH will help in characterizing alien chromosome segments captured in the durum complement and in their location in the A and/or B genome, thereby accelerating chromosome engineering research.     

The production of alien monosomic additions in Beta vulgaris as a source for the introgression of resistance to beet root nematode (Heterodera schachtii) from Beta species of the section Patellares

Summary Experiments were carried out for adding the chromosome carrying resistance to beet root nematode (Heterodera schachtii) from the wild Beta species of the section Patellares (B. procumbens, B. webbiana and B. patellaris) to the genome of B. vulgaris. Preliminary experiments indicated that crosses between the wild species and B. vulgaris cultivars of the mangold type yielded on average more viable F₁ hybrids than crosses with sugar and fodderbeet. However, crossability varied strongly between individual parental combinations. It was concluded that most types of B. vulgaris can be hybridized with the wild species of the section Patellares if a sufficient number of pair-crosses is made. Crosses between diploid cultivars or species of the section Vulgares and diploid wild species of the section Patellares yielded many hybrids which, however, were highly sterile. From crosses between tetraploid B. vulgaris and the wild species a great number of viable allotriploid and allotetraploid hybrids was obtained. In the backcross progenies of allotriploid hybrids 26% alien monosomic additions occurred, of which 4.1% carried the resistance bearing chromosome of B. procumbens or B. patellaris. The programme will be continued by sereening progenies of the resistant monosomic addition plants for the occurrence of resistant disomic introgression products.     

Wheat chromosome engineering at the 4x level: the potential of different alien gene transfers into durum wheat

Summary With the aim of making the point on feasibility and relative success of alien transfers into durum wheat via chromosome engineering, three transfer works, differing in origin and content of the alien introduction and in the transfer strategy adopted, are described. For the transfer of a powdery mildew resistance gene, Pm13, originating from Aegilops longissima and previously transferred to common wheat chromosome 3B, as well as for that of the leaf rust resistance gene Lr19 and its associated Yp (yellow pigment) gene, deriving from Ag. elongatum and introduced into 7A, the common wheat recombinants were employed as donors, from which the alien segments were homologously transferred into durum genotypes. On the other hand, for the transfer of common wheat chromosome ID seed storage protein genes, ph1 mediated homoeologous recombination was repeatedly induced. This resulted in loss of individuals, including potentially desirable recombinants, probably due to imbalances created by the ph1 condition. However, recovered Gli-D1/Glu-D3 tetraploid recombinants exhibited normal transmission and fertility. Preliminary evidence indicates a normal behaviour also for Glu-D1 5+10 putative recombinants. Similarly, there was no negative impact from the transfer of the Pm13 gene, which has been successfully pyramided into Pm4a durum varieties. On the contrary, transfer of the Ag. elongatum segment showed normal female but almost no male transmission in one durum genotype. This in spite of the fact that the alien segment, proved to be, through in situ hybridization, considerably longer than previously believed, should contain an Sd-1 gene, causing preferential transmission in common wheat. While its behaviour is being checked in other durum genotypes, shortening of the alien segment, through ph1 induced recombination, is also being carried out. Possible causes of the severe negative selection that this alien transfer seemingly encounters at the tetraploid level are discussed.     

Establishment and characterization of Brassica juncea–Orychophragmus violaceus additions, substitutions and introgressions

Intergeneric hybridizations between cultivated Brassica species and Orychophragmus violaceus have been shown to be an efficient approach to produce Brassica aneuploids. Herein B. juncea–O. violaceus additions, substitutions and introgressions were obtained among progenies of partial hybrids after B. juncea (2n = 36) was pollinated by O. violaceus (2n = 24) and they expressed some traits from O. violaceus or novel for two parents. The single O. violaceus chromosome which was added to or substituted into the B. juncea chromosome complement was distinguishable most easily in pollen mother cells at anaphase I (AI) from its darker staining and more condensed form. The one pair of the O. violaceus chromosome in the substitutions paired and segregated regularly with others from B. juncea, and the single one formed one bivalent with one B. juncea chromosome and showed normal segregation and was more darkly stained in some AI group. Stable introgressions with 2n = 36 gave amplified fragment-length polymorphism (AFLP) profiles mainly similar to those of the female B. juncea parent, but fragments specific for O. violaceus could be found in every plants together with those deleted in B. juncea and novel bands for two parents. The mechanisms behind these unusual results are discussed.