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     

Production,morphology, and cytogenetic analysis of Elymus caninus (Agropyron caninum) x Triticum aestivum F1 hybrids and backcross-1 derivatives

Summary Intergeneric hybrids were produced between common wheat, Triticum aestivum (2n=6x=42, AABBDD) and wheatgrass, Etymus caninus (Agropyron caninum) (2n=4x=28, SSHH) — the first successful report of this cross. Reciprocal crosses and genotypes differed for percent seed set, seed development and F₁ hybrid plant production. With E. caninus as the pollen parent, there was no hybrid seed set. In the reciprocal cross, seed set was 23.1–25.4% depending upon wheat genotype used. Hybrid plants were produced only by rescuing embryos 12–13 days post pollination with cv Chinese Spring as the wheat parent. Kinetin in the medium facilitated embryo germination but inhibited root development and seedling growth. The hybrids were vigorous, self sterile, and intermediate between parents. These had expected chromosome number (2n=5x=35, ABDSH), very little chromosome pairing (0.51 II, 0.04 III) and some secondary associations. The hybrids were successfully backcrossed with wheat. Chromosome number in the BC₁ derivatives varied 54–58 with 56 as the modal class. The BC₁ derivatives showed unusually high number of rod bivalents or reduced pairing of wheat homologues. These were sterile and BC₂ seed was produced using wheat pollen.     

New hybrids between Agropyron and wheat

Summary Intergeneric hybrids of Triticum aestivum (2n=42,AABBDD) with Agropyron ciliare (2n= 28,SSYY), A. trachycaulum (2n=28,SSHH), A. yezoense (2n=28,SSYY) and A. scirpeum (2n=28) are reported for the first time. F₁ hybrids of T. aestivum were also produced with A. intermedium (2n=42,E₁E₁E₂E₂Z₁Z₁) and A. junceum (2n=14,JuJu). All wheat-Agropyron hybrids were obtained by embryo rescue technique. Cultivars and reciprocal crosses differed for seed set, seed development and F₁ plant production. The F₁ hybrids were sterile. Attempts to obtain amphiploids were unsuccessful. However, backcross derivatives were obtained with wheat as the recurrent parent.The level of chromosome pairing in A. trachycaulum x wheat, A. yezoense x wheat and wheat x A. junceum hybrids provided no evidence of homologous or homoeologous pairing. Mean pairing frequencies in A. ciliare x wheat, wheat x A. scirpeum and wheat x A. intermedium hybrids indicated homoeologous or autosyndetic pairing. Ph gene was more effective in regulating homoeologous pairing in A. yezoense x wheat hybrids than in A. ciliare x wheat hybrid. Chromosome pairing data of BC₁ derivatives indicated that either some of the wheat chromosomes were eliminated or Agropyron chromosomes caused reduced pairing of wheat homologues.Contribution No. 82-653-J, Department of Plant Pathology, Kansas State Agricultural Experiment Station, Manhattan, Kan, USA     

Morphology and cytology of intergeneric hybrids involvingLeymus multicaulis (Poaceae)

The morphology and meiotic behaviour of pollen mother cells were studied in hybrids involvingLeymus multicaulis (2n = 28) ×Psathyrostachys huashanica (2n = 14),L. multicaulis ×P. juncea (2n = 14), andL. secalinus (2n = 28) ×L. multicaulis. Chromosome pairing was almost identical in theL. multicaulis ×P. huashanica, andL. multicaulis ×P. juncea hybrids, in which it averaged 7.30 univalents + 6.69 bivalents + 0.096 trivalents and 7.48 univalents + 6.75 bivalents, respectively. The meiotic pairing in the two hybrids indicated that oneL. multicaulis genome was closely homologous with theP. huashanica andP. juncea genomes. BothP. huashanica andP. juncea are possibly donors of oneL. multicaulis genome. Chromosome pairing in theL. secalinus ×L. multicaulis hybrid averaged 4.49 univalents + 11.71 bivalents + 0.02 trivalents, indicating that the genomes ofL. multicaulis andL. secalinus are to some degree homologous. However, they are sufficiently differentiated to insure species distinctness.     

Cytogenetics of a Hordeum vulgare x Elymus patagonicus hybrid (n=4x=28)

Summary Hordeum vulgare L. (2n=2x=14) was hybridized with Elymus patagonicus Speg. (2n=6x=42). The hybrid had 28 chromosomes, genomically represented as HSH₁H₂, and was perennial with a codominant phenotype. The chromosomes were meiotically associated as 19.6 univalents + 0.004 ring bivalents + 2.6 rod bivalents + 0.8 trivalents + 0.14 quadrivalents in 1,129 meiocytes, with a chiasma frequency of 4.77 per cell. The bivalent pairing presumably is an autosyndetic but modified expression of the H₁H₂ genomes of E. patagonicus, since ring bivalents were rare. This does not preclude the association of the H. vulgare H genome chromosomes with either H₁ and/or H₂ genomes of E. patagonicus to form bivalent or multivalent associations. A further evaluation of the genome homologies of H. vulgare, H. bogdanii, E. canadensis and E. patagonicus is proposed.     

Cytology of F1 hybrids and chromosome number of F2 and BC1 progeny of the cross Bromus riparius x B. inermis

Summary Hybrids between B. inermis Leyss (2n=8x=56) and B. riparius Rehm. (2n=10x=70) were easily made. The F₁ hybrids had a fertility of 20%–50% under open pollination and backcrossing to B. inermis. Chromosome pairing in B. riparius was predominantly as bivalents (29.04–33.85 per cell for plant means). Bivalents also predominated in the F₁ hybrid (2n=9x=63) and there was a high level of pairing with no reduction in chiasma frequency. It was impossible to estimate the frequency of auto-versus allosyndetic pairing. Chromosome pairing in a hybrid between B. arvensis (2n=2x=14) and B. riparius confirmed that the B. riparius complement is capable of complete autosyndetic pairing. Chromosome numbers in the F₂ progeny ranged from 2n=56 to 72 but they were skewed towards 2n=63 to 70. Backcrosses ranged from 2n=56 to 63, as expected, with the distribution skewed towards 2n=56. Selection towards the 2n=56 level would be difficult in the F₂. Empirical observation suggested that cytoplasm had a major influence on morphology in the backcrosses. Additional studies are required to determine the best breeding scheme to introgress germ plasm between B. inermis and B. riparius.     

Cytogenetic studies in the genus Zea

Summary New cytological evidence supporting x = 5 as the basic chromosome number of the genus Zea has been obtained as a consequence of our analysis of the meiotic configurations of Zea mays ssp. mays, Z. diploperennis, Z. perennis and of four F₁ artificial interspecific hybrids. Z. mays ssp. mays (2n = 20) presents regular meiosis with 10 bivalents (II) and is considered here as a typical allotetraploid (A₂A₂B₂B₂). In Z. diploperennis (2n = 20) 10II are formed in the majority of the cells, but the formation of 1III + 8II + 1I or 1III + 711 + 3I in 4% of the cells would indicate its segmental allotetraploid nature (A₁A₁B₁B₁). Z. perennis (2n = 40) had 5IV + 10II in 55% of the cells and would be considered as an auto-allooctoploid (A₁A₁A'₁A'₁C₁C₁C₂C₂). Z. diploperennis x Z. mays ssp. mays (2n = 20) presents 10II in ca. 70% of the cells and no multivalents are formed. In the two 2n = 30 hybrids (Z. mays ssp. mays x Z. perennis and Z. diploperennis x Z. perennis) the most frequent meiotic configuration was 5III + 5II + 5I and in 2n = 40 hybrid (Z. diploperennis x Z. perennis) was 5IV + 10II. Moreover, secondary association was observed in the three abovementioned tetraploid taxa (2n = 20) where one to five groups of two bivalents each at diakinesis-metaphase I was formed showing the affinities between homoeologous genomes. The results, as a whole, can be interpreed by assuming a basic x = 5 in this polyploid complex. The main previous contributions that support this working hypothesis are reviewed and its phylogenetic implications studied are discussed.     

大鹅观草与蛇河披碱草属间杂种的细胞遗传学研究

为探讨大鹅观草(Roegneria grandis,2n=4x=28)的染色体组组成,为其正确的分类处理提供细胞学依据。该研究通过人工远缘杂交,成功获得3株大鹅观草与蛇河披碱草(Elymus wawawaiensis,2n=4x=28)属间杂种F1植株。杂种植株形态介于两亲本之间,不育。亲本及杂种经I2-IK溶液染色后进行花粉育性检测,结果显示Roegneria grandis和Elymus wawawaiensis的花粉可育,育性高达94.6%和90.5%;杂种F1不育。花粉母细胞减数分裂中期I染色体配对结果显示,亲本花粉母细胞配对正常,均形成14个二价体,以环状二价体为主,Roegneria grandis有频率很低(0.04/细胞)的单价体出现;杂种F1平均每个花粉母细胞形成6.46个二价体,变化范围为5~8;在观察的83个花粉母细胞中,有35.2%的花粉母细胞形成了7个二价体,形成6个二价体的细胞占42.59%,较少细胞形成8个二价体;平均每个细胞形成14.66个单价体,变化范围为10~18;平均每细胞观察到0.14个三价体;杂种花粉母细胞染色体构型为14.66 I+6.46 II+0.14 III;平均每细胞交叉数为9.83,C值为0.35。结果表明:(1)R.grandis与Elymus wawawaiensis有一组染色体组同源的St染色体组,另外一组染色体组不是St或者H染色体组,Roegneria grandis的染色体组组成不是St Stg;(2)较低频率的三价体(平均0.14个/细胞),可能是由于R.grandis的St和Y染色体组间具有一定的同源性,也可能是染色体易位等原因导致,对于Y染色体组的起源还需深入地研究;(3)在不同地理来源的披碱草属和鹅观草属物种中St染色体组同源性不同,R.grandis与来自于北美的Elymus lanceolatus与E.wawawaiensis的St染色体组较与分布于亚洲的E.sibiricus和E.caninus的St染色体组同源性反而更高,其原因还需要进一步地研究。     

A Moricandia arvensis– based cytoplasmic male sterility and fertility restoration system in Brassica juncea

 A cytoplasmic male-sterility system has been developed in mustard (Brassica juncea) following repeated backcrossings of the somatic hybrid Moricandia arvensis (2n=28, MM)+B. juncea (2n=36, AABB), carrying mitochondria and chloroplasts from M. arvensis, to Brassica juncea. Cytoplasmic male-sterile (CMS) plants are similar to normal B. juncea; however, the leaves exhibit severe chlorosis resulting in delayed flowering. Flowers are normal with slender, non-dehiscent anthers and excellent nectaries. CMS plants show regular meiosis with pollen degeneration occurring during microsporogenesis. Female fertility was normal. Genetic information for fertility restoration was introgressed following the development of a M. arvensis monosomic addition line on CMS B. juncea. The additional chromosome paired allosyndetically with one of the B. juncea bivalents and allowed introgression. The putative restorer plant also exhibited severe chlorosis similar to CMS plants but possessed 89% and 73% pollen and seed fertility, respectively, which subsequently increased to 96% and 87% in the selfed progeny. The progeny of the cross of CMS line with the restorer line MJR-15, segregated into 1 fertile : 1 sterile. The CMS (Moricandia) B. juncea, the restorer (MJR-15), and fertility restored F₁ plants possess similar cytoplasmic organellar genomes as revealed by ‘Southern’ analysis. Received: 17 September 1997 / Accepted: 18 February 1998     

Morphological, yield, cytological and molecular characterization of a bread wheat X tritordeum F1 hybrid

The morphological, yield, cytological and molecular characteristics of bread wheat X tritordeum F₁ hybrids (2n =6x = 42; AABBDH^ch) and their parents were analysed. Morphologically, these hybrids resembled the wheat parent. They were slightly bigger than both parents, had more spikelets per spike, and tillered more profusely. The hybrids are self-fertile but a reduction of average values of yield parameters was observed. For the cytological approach we used a double-target fluorescencein situ hybridization performed with total genomic DNA fromHordeum chilense L. and the ribosomal sequence pTa71. This technique allowed us to confirm the hybrid nature and to analyse chromosome pairing in this material. Our results showed that the expected complete homologous pairing (14 bivalents plus 14 univalents) was only observed in 9.59% of the pollen mother cells (PMCs) analysed. Some PMCs presented autosyndetic pairing of Hch and A, B or D chromosomes. The average number of univalents was higher in the wheat genome (6.8) than in the Hch genome (5.4). The maximum number of univalents per PMC was 20. We only observed wheat multivalents (one per PMC) but the frequency of trivalents (0.08) was higher than that of quadrivalents (0.058). We amplified 50 RAPD bands polymorphic between the F₁ hybrid and one of its parents, and 31 ISSR polymorphic bands. Both sets of markers proved to be reliable for DNA fingerprinting. The complementary use of morphological and yield analysis, molecular cytogenetic techniques and molecular markers allowed a more accurate evaluation and characterization of the hybrids analysed here.     

Inflorescence culture of wheat-Agropyron hybrids: Callus induction,plant regeneration,and potential in overcoming sterility barriers

Segments of young inflorescences of Triticum aestivum cv. Chinese Spring (CS), its F₁ hybrids with Agropyron trachycaulum and A. scirpeum and backcross derivatives with A. yezoense, A. intermedium and A. junceum, and of a A. yezoense x T. aestivum cv. Wichita hybrid were cultured. Different parts of young spikelets of A. trachycaulum x CS F₁ and A. yezoense x Wichita F₁ 's were also cultured. Percent callus induction was lower in wheat than in the wheat-Agropyron hybrids or backcross derivatives. Percent callus induction from different organs in both hybrids was in the descending order of whole spikelet, spikelet without glumes, rachis, and glumes. No plants could be regenerated from calli of wheat and backcross derivatives except those of CS x A. intermedium combination. Callus induction in hybrids varied from 54 to 84% and plant regeneration from 14 to 31%. The regenerants required no vernalization. Variants including one with top-dense spikes and another with elongated spikelets were recovered. Out of eight A. trachycaulm x CS hybrid regenerants, one had anthers and stigma as opposed to neutral flowers of the original hybrid.     

Hybridization between Triticum aestivum L. and Agropyron michnoi Roshev.

Summary Intergeneric hybrids between Triticum aestivum cv Chinese Spring (2n=6x=42, AABBDD) and Agropyron michnoi Roshev. (2n=4x=28, PPPP) were obtained by embryo culture. Their spike characteristics were similar to those of common wheat but, unlike their parents, they were long-awned. The average meiotic chromosome pairing at MI of F₁ hybrids was: 6.39 I +3.75 rodII+8.64 ringII+0.81 III+0.30 IV+0.04 V, the bivalent and multivalent formation of which was much higher than expected from the genomic formulae. It is especially worthwhile to note that the F₁ hybrids were self-fertile, self set being 0.15%, and seeds were easily obtained from the backcross of f₁ plants with hexaploid and tetraploid wheats; here the seed set was more than 20.0%. The polyploid taxa and the position of A. Michnoi in Agropyron are discussed.     

Diplotaxis catholica + Brassica juncea somatic hybrids: molecular and cytogenetic characterization

Summary Intergeneric somatic hybrids Diplotaxis catholica (2n=18) + Brassica juncea (2n=36) were produced by fusing mesophyll protoplasts of the former and hypocotyl protoplasts of the latter using polyethylene glycol. Out of 52 somatic embryos, 24 produced plants of intermediate morphology. Cytological analysis of 16 plants indicated that 15 were symmetric hybrids carrying 54 chromosomes, the sum of the parental chromosome numbers. One hybrid was asymmetric with 45 chromosomes. Nuclear hybridity of five putative hybrids was confirmed by the Southern hybridization pattern of full length 18s-25s wheat nuclear rDNA probe which revealed the presence of Hind III fragments characteristic of both the parental species. The hybridization pattern of mitochondria specific gene probe cox I indicated that three of the hybrids carried B. juncea mitochondria and one carried mitochondria of D. catholica. Presence of novel 3.5 kb Hind III and 4.8 kb Bgl II fragments suggested the occurrence of mtDNA recombination in one of the hybrids. The hybrids were pollen sterile. However, seeds were obtained from most of the hybrids by back crossing with B. juncea.     

Cytogenetic studies on an intergeneric hybrid betweenAgropyron tsukushiense andElymus mollis

Elymus mollis is distributed widely from Korea to Japan, Kamchatka and Alaska, the northern part of U.S.S.R., and Northern and Eastern Canada, Greenland and Iceland. This species is tetraploid (2n=28). A strain of this species collected in Hokkaido was crossed withAgropyron tsukushiense var.transiens collected in Mishima. From this cross, 22 F₁ plants were produced. Crossability calculated from the number of hybrid plants produced and the number of floret pollinated was 30.6%. The shape of the F₁ spikes was of theAgropyron type but the glumes were hairy as were those of theElymus parent. One of the characteristics distinguishingElymus fromAgropyron is the production of two spikelets at almost all nodes of the rachis. This character was not expressed in the F₁ plants. All pollen grains of the F₁ plants were completely abortive. The average chromosome pairing at the MI of the PMCs of the F₁ amounted to 2.03 bivalents and 30.95 univalents. Almost all bivalents ranging from one to seven were rod-shaped connected with interstitial or terminal chiasma. These results indicate a lack of genomic homology between the three genomes ofA. tsukushiense and the two genomes ofE. mollis. Contribution No. 37 from the Plant Germ-plasm Institute, Faculty of Agriculture, Kyoto University, Kyoto, Japan.     

Introgression of chromosomes of Festuca arundinacea var. glaucescens into Lolium multiflorum revealed by genomic in situ hybridisation (GISH)

An F₁ hybrid (n=4x=28) between the tetraploid species Festuca arundinacea var. glaucescens (GGG′G′) and a synthetic tetraploid Lolium multiflorum (LmLmLmLm) was backcrossed to diploid L. multiflorum to produce triploid (2n=3x=21) BC₁ hybrids (LmLmG). At metaphase I of meiosis the triploids had a preponderance of ring bivalents and univalents with some linear and frying-pan trivalents. Genomic in situ hybridisation (GISH) differentiated the Festuca chromosomes from Lolium and revealed that the bivalents were exclusively between Lolium homologues, while the univalents were Festuca. Despite the limited amount of homoeologous chiasmata pairing in the triploids, some recombinant chromosomes were recovered in the second backcross when the hybrids were further crossed to diploid L. multiflorum. The progeny from the second backcross was predominantly diploid. Genotypes with recombinant chromosomes and chromosome additions involving an extra Festuca chromosome were identified using GISH. Changes in plant phenotype were related to the presence of Festuca chromatin. Received: 20 September 2000 / Accepted: 05 January 2001     

Intergeneric hybridization between Diplotaxis siifolia,a wild species and crop brassicas

Summary Attempts were made to obtain intergeneric hybrids between Diplotaxis siifolia, a wild species, and cultivars of Brassica (B. campestris, B. juncea, and B. napus). The crosses showed unilateral incompatibility. When the wild species was used as female parent, pollen germination and pollen tube growth were normal, but hybrid seeds aborted due to post-fertilization barriers. Reciprocal crosses (cultivars as female parent) showed strong pre-fertilization barriers; although pollen grains showed germination, pollen tubes failed to enter the stigma. Hybrids were realized in two of the crosses, D. siifolia x B. juncea and D. siifolia x B. napus, through ovary culture. The hybrids were multiplied in vitro by multiplication of axillary shoots, or somatic embryogenesis. Detailed studies were carried out on the hybrid D. siifolia x B. juncea. F₁ hybrids had shrivelled anthers and were pollen sterile. Amphiploids of this hybrid showed 60% pollen fertility and produced seeds upon self-pollination as well as backcross pollination with the pollen of B. juncea.     

GISH and AFLP analyses of novel Brassica napus lines derived from one hybrid between B. napus and Orychophragmus violaceus

New Brassica napus inbred lines with different petal colors and with canola quality and increased levels of oleic (∼70%, 10% higher than that of B. napus parent) and linoleic (28%) acids have been developed in the progenies of one B. napus cv. Oro × Orychophragmus violaceus F₅ hybrid plant (2n=31). Their genetic constituents were analyzed by using the methods of genomic in situ hybridization (GISH) and amplified fragments length polymorphism (AFLP). No intact chromosomes of O. violaceus origin were detected by GISH in their somatic cells of ovaries and root tips (2n=38) and pollen mother cells (PMCs) with normal chromosome pairing (19 bivalents) and segregation (19:19), though signals of variable sizes and intensities were located mainly at terminal and centromeric parts of some mitotic chromosomes and meiotic bivalents at diakinesis or chromosomes in anaphase I groups and one large patch of chromatin was intensively labeled and separated spatially in some telophase I nuclei and metaphase II PMCs. AFLP analysis revealed that substantial genomic changes have occurred in these lines and O. violaceus–specific bands, deleted bands in ‘Oro’ and novel bands for two parents were detected. The possible mechanisms for these results were discussed.     

Somatic hybridization between Brassica juncea and Moricandia arvensis by protoplast fusion

Intergeneric somatic hybrids have been produced between Brassica juncea (2n=36, AABB) cv. RLM-198 and Moricandia arvensis (2n=28, MM) by protoplast fusion. Hypocotyl protoplasts of B. juncea were fused with mesophyll protoplasts of M. arvensis using polyethylene glycol. Fusion frequency, estimated on the basis of differential morphological characterstics of parental protoplasts was about 5%. Of the 156 calli obtained, four calli produced shoots intermediate in morphology between the parents. Hybrid nature of the plants was confirmed using wheat nuclear rDNA probe. Hybridization of total DNA with a mitochondrial DNA probe carrying 5s–18s rRNA genes of maize showed that the mitochondria of the somatic hybrids were derived from the wild species M. arvensis. Meiosis in the only hybrid that produced normal flowers revealed the occurrence of 64 chromosomes, the sum of chromosomes of parental species. Inspite of complete pollen sterility, siliquas were produced in this hybrid by back-crossing with B. juncea. These siliquas on in vitro culture produced 12 seeds.     

Production,morphology, and cytogenetics of Triticum aestivum (L.) Thell × Elymus scabrus (R. Br.) Love intergeneric hybrids obtained by in ovulo embryo culture

Summary Intergeneric hybrids were produced between common wheat, Triticum aestivum (2n=6x=42, AABBDD), and an apomictic Triticeae species, Elymus scabrus (syn. Agropyron scabrum) (2n=6x=42, HHSSSS), the first successful report of this cross. Nine tiny, underdeveloped, and structureless embryos were obtained in vitro only by in ovulo embryo culture at 4 days after pollination, which gave rise to five mature hybrid plants. All the hybrid plants were vigorous and possessed a phenotype intermediate to the two parents. There were 2n=6x=42 (ABDHSS) somatic chromosomes in the hybrids. There was little or no homology between the parental genomes, as shown by an overall meiotic chromosome association of 32.83 I + 4.08 rod II + 0.21 ring II + 0.18 III + 0.02 IV. The hybrids were completely sterile and so far backcrosses to wheat parent have not been successful. Alternate approaches to induce gene transfer(s) from E. scabrus to wheat are being attempted.Agriculture Canada Contribution No. 398.     

A study of morphology,cytology and sterility in interspecific hybrids and amphidiploids of Nicotiana knightiana X N. umbratica

Summary Interspecific hybrids and amphidiploids of Nicotiana knightiana Goodspeed (n= 12)x N. umbratica Burbidge (n = 23) resembled either parent in some characters and were intermediate in other characters. The F₁ hybrids (2n = 35) showed mostly univalents during meiosis, while the amphidiploids (2n = 70) formed bivalents almost regularly. The former were completely sterile and the latter fully male fertile but predominantly female sterile. This female sterility was due to disintegration of the embryo sacs leading to collapsed ovules. The few fertile ovules, however, showed normal development of embryo sac and embryo. The occurrence of fertile and sterile ovules was believed to be due to segregation of the genes governing sterility.