Identification of wheat-Agropyron cristatum monosomic addition lines by RFLP analysis using a set of assigned wheat DNA probes

A non-radioactive digoxigenin-labelled DNA method was used successfully to identify RFLP markers in 54 Triticum aestivum cv Chinese Spring — Agropyron cristatum (2n=28, genome PPPP) P-genome monosomic addition lines. Southern analysis using a set of 14 DNA probes identifying each homoeologous chromosome arm, combined with two restriction enzymes HindIII and EcoRI, indicated that six A. cristatum chromosomes (1P, 2P, 3P, 4P, 5P and 6P) and five A. cristatum chromosome arms (2PS, 2PL, 5PL, 6PS and 6PL) have been individually added to the wheat genome. The added chromosomes of three lines were Agropyron translocated chromosomes. It was also found that two addition plants possessed an Agropyron-wheat translocation. These results showed that RFLP analysis using the set of assigned wheat probes was a powerful tool in detecting and establishing homoeology of alien A. cristatum chromosomes, or arms, added to wheat, as well as in screening the alien addition material. The creation of the monosomic addition lines should be useful for the transfer of disease-resistance genes from A. cristatum to wheat.     

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.     

Selection and identification of a spontaneous alien chromosome translocation in wheat

A wheat (Triticum aestivum L. emend Thell) disomic addition line (2n = 6x = 44), SH1–152–2, with a pair of Elytrigia pontica (Podp.) Holub 2n = 10x = 70 [syn. Agropyron elongatum (Host) P.B.] chromosomes controlling blue aleurone color was crossed with a short-statured spring wheat `Sonora 64' (T. aestivum). Isoline pairs of blue-disomic addition lines and nonblue euploid lines were produced by selecting plants segregating for blue aleurone for 12 generations. Nineteen of 20 blue aleurone lines were 2n = 44 addition lines, and one had 2n = 42 chromosomes. Several lines of evidence showed that this line had a spontaneous translocation in which the β arm of wheat chromosome 4A was replaced by an Elytrigia chromosome arm carrying the blue aleurone gene. The Elytrigia chromosome in SH1–152–2 appeared to be homologous with E. pontica chromosome 4el₁, which also carries the blue aleurone gene. It was concluded that the spontaneous translocation originated from simultaneous misdivision of univalents and subsequent reunion at the centromere of chromosome arm 4Aα with the Elytrigia chromosome arm.     

Utilization of blue-grained character in wheat breeding derived from Thinopyrum poticum

The blue grain trait in common wheat （Triticum aestivum L., 2n = 6x = 42, AABBDD）, which is caused by blue pigments in the aleurone layer, was originally derived from the tall wheatgrass （Thinopyrum ponticum Liu ＆ Wang = Agropyron elongatum, 2n = 10x = 70, StStStStEeEeEbEbEXEx） during chromosome engineering research. Over the last few decades, there have been continued interests in the genetic mechanism of this blue coloration and the practical utilization of the blue aleurone character as a phenotypic marker. This article reviews the research history and the recent progress of the studies on blue-grained wheat, with emphases on genetic and biochemical analysis and practical applications of blue-grained wheat.     

Haploidy from Hordeum interspecific crosses

Summary Hordeum arizonicum (2n=42) and H. lechleri (2n=42) were crossed with both H. bulbosum (2n=14 or 28) and H. vulgare (2n=14 or 28) and progeny plants were obtained through embryoculture. Crosses of arizonicum with diploid bulbosum invariably resulted in haploids (2n=21) of arizonicum, whereas arizonicum by tetraploid bulbosum or diploid vulgare crosses produced both hybrids and haploids of arizonicum. The lechleri by diploid bulbosum or diploid vulgare crosses resulted in haploids of lechleri, while lechleri by tetraploid bulbosum resulted in well differentiated embryos which failed to germinate.Hybrid embryos derived from the haploid producing crosses exhibit chromosome variability, suggesting that chromosome elimination leads to haploid formation.The results also indicate that the ratio of the parental genomes in the zygote is a critical factor which determines the chromosome elimination or stability in any cross combination. Furthermore, both arizonicum and lechleri appear to be of similar genetic strength in eliminating bulbosum and vulgare chromosomes. The possibility of stability factors in overcoming elimination and manipulation towards elimination are discussed.     

The genome specific grain proteins and the phylogenetic interrelation between Triticum L., Elytrigia Desf., Elymus L. and Agropyron Gaertn

Summary Immunochemical protein studies show that nongliadins of the ethanol-soluble fraction (EF) provide the best biochemical information on genome interrelations in cereals. After electrophoresis (pH 3.2) these genome specific proteins are shown to fall within the category of -prolamines and albumins where inhibitors against -amylases are to be found. Although these genome specific proteins of the EF exhibit identical serological properties, they may differ in electrophoretic mobility. They may very well be controlled by different chromosomes within a certain, characteristic genome. Electrophoretic analyses will thus likely reveal interrelations between individual chromosomes or even chromosome segments while the precipitation spectrum of the EF will function as a marker of the whole genomes or at least of the major part of their genetic material. This advantage of the immunological technique has been explored in a study of the phylogenetic relations between different genomes belonging to Triticum L., Elytrigia Desf., Elymus L. and Agropyron Gaertn. Serological markers for the main genomes of Triticum were checked against species and genomes of the other three genera.More than 80 species belonging to these three genera were proved to be immunologically distinctive from the wheat genomes. Antigens produced against diploid Elytrigia species were not only checked with other species belonging to Elytrigia but also to Elymus and Agropyron. This inventory of interrelationships confirmed previous knowledge and also added some new information on the phylogeny of the polyploid representatives of Elytrigia and Elymus.     

Messenger RNA from isolated aleurone cells directs the synthesis of an alpha-galactosidase found in the endosperm during germination of guar (Cyamopsis tetragonaloba) seed

In cereals and some legumes the aleurone layer is a site of synthesis of enzymes which mobilize endosperm reserves. It has been established whether or not the aleurone cells of the seed endosperm of Cyamopsis tetragonaloba are a site of synthesis of -galactosidase. The isolation and cultivation of aleurone cells demonstrated that they contain mRNA which directs the synthesis and secretion of -galactosidase into the endosperm where along with a -mannanase it is responsible for the degradation of the galactomannan storage polymer. A method was developed to purify the mRNA from the aleurone cells of germinating seeds. This mRNA was analysed by: i) Northern blot hybridization using oligo-nucleotide mixed probes derived from the protein's NH₂-terminal amino acid sequence and ii) in vitro translation in a wheat germ system and detection of the -galactosidase protein using antibodies. The molecular mass of the protein synthesized in vitro is slightly larger (44 kDa) than that of the mature -galactosidase (40.5 kDa) which is as expected for the precursor of a secreted protein.     

Chromosomal location of genes controlling seed proteins in species related to wheat

Summary The seed proteins of Chinese Spring wheat stocks which possess single chromosomes from other plant species related to wheat have been separated by gel electrophoresis in the presence of sodium dodecyl sulphate. Marker protein bands have been detected for both arms of barley chromosome 5, chromosome E (= 1R) and B (= 2R) of rye, chromosomes A,B (= 1C^u) and C (= 5C^u) of Aegilops umbellulata and chromosomes I and III of Agropyron elongatum. These studies, and previous findings, indicate that chromosome 5 of barley, chromosome 1R of rye, chromosome I of Ag. elongatum and possibly chromosome 1C^u of Ae. umbellulata are similar to chromosomes 1A, 1B and 1D in hexaploid wheat in that they carry genes controlling prolamins on their short arms and genes controlling high-molecular-weight (apparent molecular weight greater than 86,000) seed protein species on their long arms. These findings support the idea that all these chromosomes are derived from a common ancestral chromosome and that they have maintained their integrity since their derivation from that ancestral chromosome.     

Identification of Haynaldia villosa chromosomes added to wheat using a sequential C-banding and genomic in situ hybridization technique

Genomic in situ hybridization (GISH) offers a convenient and effective method for cytological detection, but can not determine the identity of the chromosomes involved. We integrated C-banding with GISH to identify Haynaldia villosa chromosomes in a wheat background. All chromosomes of H. villosa showed C-bands, either in telomeric regions or in both telomeric and centromeric regions, which allowed unequivocal identification of each H. villosa chromosome. The seven pairs of H. villosa chromosomes were differentiated as 1–7 according to their characteristic C-bands. Using a sequential C-banding and GISH technique, we have analyzed somatic cells of F₃ plants from the amphiploid Triticum aestivum-H. villosa x Yangmai 158 hybrids. Three plants (94009/5-4,94009/5-8 and 94009/5-9) were shown to contain H. villosa chromosome(s). 94009/5-4 (2n = 45) had three H. villosa chromosomes (2, 3 and 4); 94009/5-8 (2n = 45) possessed one chromosome 4 and a pair of chromosome 5, and 94009/5-9 (2n = 43) was found to have one chromosome 6 of H. villosa. The combination of GISH with C-banding described here provides a direct comparison of the cytological and molecular landmarks. Such a technique is particularly useful for identifying and localizing alien chromatin and DNA sequences in plants.     

Production and cytogenetic analysis of BC1, BC2, and BC3 progenies of an intergeneric hybrid between Triticum aestivum (L.) Thell. and tetraploid Agropyron cristatum (L.) Gaertn.

Summary Intergeneric hybrids between Triticum aestivum cv Chinese Spring and Agropyron cristatum 4x (2n= 5x=35, ABDPP genomes) with a high level of homoeologous meiotic pairing between the wheat chromosomes were backcrossed 3 times to wheat. Pollination of the F₁ hybrid with Chinese Spring resulted in 22 BC₁ seeds with an average seed set of 1.52%. Five BC₁ plants with 39–41 chromosomes were raised using embryo rescue techniques. Chromosome pairing in the BC₁ was characterized by a high frequency of multivalent associations, but in spite of this there was no evidence of homoeologous pairing between chromosomes of wheat and those of Agropyron. All of the plants were self sterile. The embryo rescue technique was again essential to produce 39 BC₂ plants with chromosome numbers ranging from 37 to 67. The phenomenon of meiotic non-reduction was also observed in the BC₃ progenies. In this generation male and female fertility greatly increased, and meiotic pairing was fairly regular. Some monosomic (2n=43) and double monosomic (2n=44) lines were produced. Analysis of these progenies should permit the extraction of the seven possible wheat-Agropyron disomic addition lines including those with the added chromosomes carrying the genes involved in meiotic non-reduction and in suppression of Ph activity.     

A self-fertile trigeneric hybrid,Triticum aestivum ×Agropyron michnoi ×Secale cereale

Trigeneric hybrids between the (Triticum aestivum ×Agropyron michnoi) F₁ (CM, 2n=5x=35; ABDPP) and two winter rye (Secale cereale L., 2n=2x=14; RR) cultivars, Wugong 774 and AR-132, were synthesized. Such trigeneric hybrids could be used to transfer resistance genes for powdery mildew from rye to CM and subsequently to common wheat and to identify (1) the effects of the P genome ofAgropyron on the self-fertility of the hybrids and (2) the differences in genetic background between rye cultivars with marked differences in pollinating habit. The trigeneric hybrids varied widely in morphology and showed a high level of resistance to such diseases as barley yellow dwarf virus (BYDV), stripe rust, leaf rust, stem rust, and powdery mildew. Selfed and many backcross derivatives were obtained from the trigeneric hybrids. The results indicated that rye cvs Wugong 774 and AR132 arose from different gene pools and that the P genome ofAgropyron carries gene(s) responsible for chromosome segregation, leading to functional gamete formation and self-fertility of the hybrids. The F₂ and BC₁ plants could be obtained in two ways — fusion of the unreduced gametes and the assumed apomixis of unreduced female gametes in the trigeneric hybrid plant II-4 — which indicates that this trigeneric hybrid may be a special genetic stock. Chromosome pairing in the trigeneric hybrids and ways of producing wheat/rye and wheat/Agropyron translocations are discussed.     

Production and cytogenetics of Triticum aestivum L. hybrids with some rhizomatous Agropyron species

Summary Intergeneric hybrids between Triticum aestivum L. and conventional rhizomatous Agropyron species were produced in variable frequencies. They were recovered in high percentage frequencies for T. aestivum cultivars with A. acutum (14.6%), A. intermedium (48.0%), A. pulcherrimum (53.3%), and A. trichophorum (46.6%). The crossability percentages with the highly crossable cultivar Chinese Spring for these Agropyron species accessions were 33.12%, 65.0%, 53.3%, and 65.4%, respectively. Autosyndetic associations of two of their three genomes gave mean meiotic chromosome association data of 17.0 I (univalents) +1.53 II (ring bivalents) + 7.04 II (rod bivalents) +1.43 III (trivalents) +0.05 IV (quadrivalents) +0.01 IV (pentavalents) for A. acutum and of 21.8 I + 1.56 II (rings) +7.22 II (rods) +0.84 III + 0.04 IV for A. intermedium. Chromosome pairing at metaphase I was comparatively lower for A. pulcherrimum (34.4 I + 0.2 II (rings) +3.4 II (rods) +0.14 III) and A. trichophorum (36.7 I + 0.35 II (rings) +2.26 II (rods) + 0.04 III) hybrids with T. aestivum. Hybrids of wheat with A. campestre and A. repens were obtained in low frequency. Direct crossing did not permit T. aestivum/ A. desertorum hybridization. However, by utilizing the 2n=10x=70 A. repens/A. desertorum amphiploid as the pollen source, hybridization with T. aestivum did indeed occur. Aneuploidy was prevalent in this hybrid combination while all other hybrid combinations were apparently normal.     

Relationships of Agropyron intermedium chromosomes determined by chromosome pairing and alcohol dehydrogenase isozymes in common wheat background

Summary The relationships of Agropyron intermedium chromosomes in two wheat-Agropyron addition series were determined. Chromosome pairing behaviour revealed that the alien chromosome in lines TAF-2 and L7 of Vilmorin-A. intermedium set are homologous to the alien chromosomes in lines P and C of the Caribo-A. intermedium set respectively. Localization of alcohol dehydrogenase isozyme genes in Vilmorin-Agropyron addition line L4 and in Caribo-Agropyron line O indicated relationships with wheat chromosomes of homoeologous group 4.     

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.     

Chromosome painting of Amigo wheat

Chromosome painting using multicolor fluorescence in situ hybridization showed that, in addition to the T1AL·1RS translocation derived from rye, a segment from chromosome 3Ae#1 of Agropyron elongatum (2n=10x =70), is present in Amigo wheat. The Agropyron chromosome segment is located on the satellite of chromosome 1B and the translocation chromosome is designated as T1BL·1BS-3Ae#1L. T1BL·1BS-3Ae#1L was inherited from Teewon wheat and carries resistance genes to stem rust (Sr24) and leaf rust (Lr24). The Agropyron chromosome segments in different Sr24/Lr24 carrier wheat lines, including Agent, TAP 48, TAP 67, Teewon, and Amigo, showed a diagnostic C-band, and were derived from the same chromosome, 3Ae#1.     

Extent of genetic variability of endosperm esterases in Triticum aestivum L. 2n=6x=42

Summary Genetic variability of endosperm esterase has been studied in 42 cultivars of Triticum aestivum L. 2n=6x=42. Different techniques, including sequential electrophoresis and electrofocusing, have been used with various substrates and esterase inhibitors. The electrophoretic patterns in each cultivar are described. Chromosomal location using the nullitetrasomic and ditelosomic lines of Chinese Spring was carried out in order to relate and/or locate the esterase genes to specific chromosomes. Most of the esterase isozymes located were in the long arm of the chromosomes of the homoeology group 3; but we have found six located in the short arms, five of them in the chromosome 3AS and one in the 3DS. This location increases the number of esterase genes described, because no esterase genes had been described so far in short arms of chromosomes of the homoeology group 3. The genetic control is discussed and, according to our results, between 12 and 15 loci, organized in five compound loci, control the endosperm esterases in wheat. Also one modifier gene modifying the mobility of two esterase bands and present in all the cultivars studied is postulated.This work was supported by a personal grant (L. Rebordinos) from the P.F.P.I. and by an institutional grant from the C.A.I.C.Y.T. (PB85-0153)     

C-banding pattern and polymorphism of Aegilops caudata and chromosomal constitutions of the amphiploid T. aestivum — Ae. caudata and six derived chromosome addition lines

Summary C-banding patterns were analysed in 19 different accessions of Aegilops caudata (= Ae. markgrafii, = Triticum dichasians) (2n = 14, genomically CC) from Turkey, Greece and the USSR, and a generalized C-banded karyotype was established. Chromosome specific C-bands are present in all C-genome chromosomes, allowing the identification of each of the seven chromosome pairs. While only minor variations in the C-banding pattern was observed within the accessions, a large amount of polymorphic variation was found between different accessions. C-banding analysis was carried out to identify Ae. caudata chromosomes in the amphiploid Triticum aestivum cv Alcedo — Ae. caudata and in six derived chromosome addition lines. The results show that the amphiploid carries the complete Ae. Caudate chromosome complement and that the addition lines I, II, III, IV, V and VIII carry the Ae. caudata chromosome pairs B, C, D, F, E and G, respectively. One of the two SAT chromosome pairs (A) is missing from the set. C-banding patterns of the added Ae. caudata chromosomes are identical to those present in the ancestor species, indicating that these chromosomes are not structurally rearranged. The results are discussed with respect to the homoeologous relationships of the Ae. caudata chromosomes.     

Intergeneric hybrids between Triticum crassum and Hordeum vulgare

Summary Intergeneric hybrids between Triticum crassum (2n=6x=42) and Hordeum vulgare cv. Bomi were obtained at a frequency of 15% of pollinated florets. Meiotic chromosome pairing in the hybrids was not different from that observed in a polyhaploid of T. crassum indicating negligible pairing between chromosomes of the two species and secondly that the genome of H. vulgare had no effect on intergenomic pairing in T. crassum.Contribution No. 646 Ottawa Research Station     

A monoclonal antibody chromosome marker analysis used to locate a loose smut resistance gene in wheat chromosome 6A

Many genes have been located in wheat chromosomes, yet little is known about the location of genes for resistance to Ustilago tritici, which causes loose smut. Crosses were made between the loose smut susceptible alien substitution lines Cadet 6Ag(6A) and Rescue 6Ag(6A) (lines in which Agropyron chromosome 6 is substituted by wheat chromosome 6A) and four cultivars resistant to U. tritici race T19: Cadet, Kota, Thatcher and TD18. The segregating progeny were tested for reaction to race T19 and for the level of binding with a monoclonal antibody specific to a chromosome 6A-coded seed protein. The antibody, which does not bind to seed protein extracts in the absence of the 6A chromosome, was used as a chromosome marker. An association was established between resistance to race T19 and the presence of chromosome 6A for each of the cultivars tested, indicating that resistance to race T19 resides in chromosome 6A. Ustilago tritici race T19 resistance in Cadet appears to be located in the short arm of chromosome 6A, based on the evaluation of the Cadet 6A long ditelosomic stock, which was susceptible, and the Cadet 6A-short: 6-Agropyron- short alien translocation stock, which was resistant.     

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