In situ hybridization as a rapid means to assess meiotic pairing and detection of alien DNA transfers in interphase cells of wide crosses involving wheat and rye

Summary The objectives of this study were to determine if biotin-labelled total genomic DNA of rye (Secale cereale L.) could be used to (i) preferentially label rye meiotic chromosomes in triticale and (ii) detect translocation stocks at interphase and/or early prophase by in situ hybridization. Welsh triticale, a wheat-rye segmental amphiploid, and Kavkaz wheat, a wheat-rye translocation were used. The results indicated that labelled chromosomes of rye and unlabelled chromosomes of wheat could be observed throughout all meiotic stages in the triticale. For Kavkaz wheat, the presence of the translocated 1RS chromosome arm of rye was detected at the interphase or very early prophase stage. Rapid assessment of feasibility of gene transfers and detection of alien DNA in somatic cells at the interphase stage by in situ hybridization allows for rapid decision-making and saves time and expense in plant breeding programs.Plant Research Centre Contribution No. 1276     

The mapping of highly-repeated DNA families and their relationship to C-bands in chromosomes of Secale cereale

The relationship between the chromosomal location of heterochromatin C-bands and of four non-homologous repeated sequence families constituting 8 to 12% of total rye DNA has been investigated in chromosomes of rye (Secale cereale) by in situ hybridisation. Three rye varieties, a set of rye disomic additions to wheat and a triticale were studied. Only centromeric and nucleolar organizer region (NOR) associated C-bands failed to display hybridisation to at least one of the sequences and many telomeric blocks of heterochromatin contained all four repeated sequence families. Both between-variety differences in the chromosomal distribution of repeated sequences, and intravarietal heterozygosities were frequently noted and are probably widespread. — Previously reported deletions of heterochromatin from King II rye chromosomes added to the Holdfast wheat complement were correlated with deletions of some, but not all, of the highly repeated sequence families. A previously unreported loss of some families from King II rye chromosome 4R/7R in a Holdfast wheat genetic background was detected. This loss was not associated with complete deletion of a C-band. A deletion has also probably occurred from the short arm telomere of 4R/7R in the triticale variety Rosner. It is suggested that the families of repeats in rye telomeric heterochromatin which are absent from wheat are selected against in the wheat genetic background.     

Structural variation in the heterochromatin of rye chromosomes in triticales

Summary Although Giemsa C-banding techniques have been used extensively for assaying cereal heterochromatin, a more specific technique for analyzing cereal heterochromatin has been developed recently with the isolation of DNA sequences present in heterochromatin and their employment in in situ hybridization to cereal chromosomes. A number of triticales were examined for the occurrence of modified rye chromosomes using the in situ hybridization technique. With a heterogeneous sequence probe the amount of rye heterochromatin appears to be relatively constant in wheat backgrounds but when a specific sequence probe was employed variation was observed. Whether this variation reflects polymorphism in rye or whether it is a result of adaption of the rye genome to coexistence with the wheat genome in triticales is discussed. — The triticale Rosner was examined in detail and it was established that the rye chromosome 2R had been replaced by the wheat chromosome 2D.     

Transmission of chromosomes through the eggs and pollen of triticale × wheat F1 hybrids

Summary The substitution patterns of rye chromosomes in hexaploid triticale × wheat F₂ hybrids, along with the transmission patterns of rye chromosomes through egg cells and pollen when several of the F₁ hybrids were test crossed to triticale and wheat were investigated. The data indicated that the rye chromosome transmission through both the egg and pollen was random in number and in composition. The test crosses suggested that it was best to use wheat pollen for the transmission of rye chromosomes through the egg cells of the F₁ hybrids and triticale egg cells for the transmission of rye chromosomes through F₁ hybrid pollen. A deviation from random segregation in the F₂ and the transmission rate was observed for rye chromosomes 1R, 4R/7R, and 6R. The transmission rates of 1R and 6R varied depending on the direction in which the cross was made. The results also indicated that there was little or no compensation between the R- and D-genomes and that the chromosomes of these two genomes appeared to be transmitted independently of each other.     

Development of commercially valuable traits in hexaploid triticale lines with <Emphasis Type="Italic">Aegilops</Emphasis> introgressions as dependent on the genome composition

Introgressive hybridization is an efficient means to improve the genetic diversity of cultivated cereals, including triticale. To identify the triticale lines with Aegilops introgressions, genotyping was carried out with ten lines obtained by crossing hexaploid triticale with genome-substitution forms of the common wheat cultivar Aurora: Aurolata (AABBUU), Aurodes (AABBSS), and Aurotika (AABBTT). The genome composition of the triticale lines was studied by in situ hybridization, and recombination events involving Aegilops and/or common wheat chromosomes were assumed for nine out of the ten lines. Translocations involving rye chromosomes were not observed. Substitutions for rye chromosomes were detected in two lines resulting from crosses with Aurolata. Genomic in situ hybridization (GISH) with Ae. umbellulata DNA and molecular genetic analysis showed that chromosome 1R was substituted with Ae. umbellulata chromosome 1U in one of the lines and that 2R(2U) substitution took place in the other line. Fluorescence in situ hybridization (FISH) with the Spelt1 and pSc119.2 probes revealed a translocation from Ae. speltoides to the long arm of chromosome 1B in one of the two lines resulting from crosses with Aurodes and a translocation in the long arm of chromosome 7B in the other line. In addition, the pSc119.2 probe revealed chromosome 1B rearrangements in four lines resulting from crosses with Aurolata and in a line resulting from crosses with Aurotika. The lines were tested for main productivity parameters. A negative effect on all productivity parameters was demonstrated for Ae. umbellulata chromosome 2U. The overwinter survival in all of the lines was similar to or even higher than in the original triticale cultivars. A substantial increase in winter resistance as compared with the parental cultivar was observed for the line carrying the T7BS-7SL translocation. The line with the 1R(1U) chromosome substitution seemed promising for the baking properties of triticale.     

Characterisation of rye B chromosome DNA by DNA/DNA hybridisation

The DNAs of wheat and rye plants with rye B chromosomes have been compared with wheat, rye and oats DNAs by DNA/DNA hybridisation. The presence of DNA from B chromosomes made no significant difference to the proportion of repeated sequence DNA. The repeated sequence fractions of these cereal DNAs were quantitatively divided into eight different groups on the basis of the amount of DNA/DNA hybridisation occurring between the different DNAs. Rye A and B chromosomes contained similar proportions of three of the groups. These results, together with estimates of the thermal stabilities of all the renatured DNA duplexes suggest that rye B chromosome DNA is very similar to rye A chromosome DNA in the proportion and heterogeneity of its repeated sequences.     

八倍体小黑麦×普通小麦杂种后代群体中的染色体易位

用改良的Giemsa C-带技术以单株为基础分析了八倍体小黑麦×普通小麦的杂种BC_1,F_(?)和F_(?)代植株的核型。在鉴定了C-带核型的1098株杂种后代植株中,发现了78条小麦-黑麦和277条黑麦-黑麦易位染色体。在不同的世代和株系中,小麦-黑麦染色体易位率变化在4.35—14.07%之间,平均7.10%;黑麦-黑麦染色体易位率在0.48—52.78%之间,平均25.23%。鉴定的小麦-黑麦易位染色体涉及了黑麦的14条不同的染色体臂和小麦的A、B和D组染色体。易位的48.57%发生在小麦和黑麦的部分同源染色体之间,51.43%发生在非部分同源染色体之间。不同的黑麦染色体臂参与易位的频率不同。小麦-黑麦染色体易位主要发生在杂种的早期世代,使用适当的选择技术在F_3获得了纯合的易位植株。文中讨论了快速选育易位系的技术和它们在小麦育种中的应用问题。     

Identification of Alien Chromatin and Ribosomal DNA in Wheat by in Situ Hybridization

In situ hybridization was carried out to somatic cells of hexaploid Triticale “Badger”, lB/IR translocation line “Ning 8026” and IR(ID) substitution line “84056-1-36-1” using biotin-labelled total rye genomic DNA and wheat rDNA as probes, the results were as follows: 1. The probe containing the total genomic DNA from rye hybridized to the entire length of all rye chromosomes, as a result of the formation of a brown precipitate over the sites of hybridization, the rye chromosomes could be distinguished from wheat chromosomes counterstained by Wright’s solution, the distinguishable appearance of the wheat and rye chromosomes resulted in an efficient method of detecting rye chromosome or segments in wheat. 2. When the probe PTA 71 containing wheat ribosomal DNA was used to hybridize to somatic chromosomes of "Badger" and “84056-1-36-1”, six signals in “Badger” and eight in “84056-1-36-1” were observed on lB, 6B, 1R and SD, among which lB and 6B showed large in situ signals corresponding to many copies of the genes. 3. The expression behavior of wheat rDNA was found in interphase cells by in situ hybridization.     

Molecular Cytogenetic Identification of Triticum aestivum-Secale cereale Substitution and Addition Lines

Two substitution lines, designated as 930498 and 930483, and one addition line, designated as 930029, via Fo immature embryo culture of Triticum aestivum x octoploid triticale ( x Triti-cosecale Wittmack) were identified. Fluorescence in situ hybridization (FISH) using total genomic DNA of rye ( Secale cereale L. ) as probe corroborated the existence of rye chromosomes, further confirmed through chromosome paring at meiotic metaphase 1, C-banding and glutenin SDS- PAGE. The results demonstrated that the two substitution lines are ID/IR, and the addition line is also IR addition. Rye chromosomes that are distinct to the red-colored wheat chromosomes appear yellow-green at mitotic metaphase after FISH.     

Painting the rye genome with genome-specific sequences

We used rye-specific repetitive DNA sequences in fluorescence in situ hybridization (FISH) to paint the rye genome and to identify rye DNA in a wheat background. A 592 bp fragment from the rye-specific dispersed repetitive family R173 (named UCM600) was cloned and used as a FISH probe. UCM600 is dispersed over the seven rye chromosomes, being absent from the pericentromeric and subtelomeric regions. A similar pattern of distribution was also observed on the rye B chromosomes, but with weaker signals. The FISH hybridization patterns using UCM600 as probe were comparable with those obtained with the genomic in situ hybridization (GISH) procedure. There were, however, sharper signals and less background with FISH. UCM600 was combined with the rye-specific sequences Bilby and pSc200 to obtain a more complete painting. With these probes, the rye chromosomes were labeled with distinctive patterns; thus, allowing the rye cultivar 'Imperial' to be karyotyped. It was also possible to distinguish rye chromosomes in triticale and alien rye chromatin in wheat-rye addition and translocation lines. The distribution of UCM600 was similar in cultivated rye and in the wild Secale species Secale vavilovii Grossh., Secale sylvestre Host, and Secale africanum Stapf. Thus, UCM600 can be used to detect Secale DNA introgressed from wild species in a wheat background.     

Wheat Neocentromeres Found in F1 Triticale × Tritordeum Hybrids (AABBRH<Superscript>ch</Superscript>) After 5-Azacytidine Treatment

The DNA hypomethylation effect of 5-azacytine (5-AC; a cytosine analog) is widely known. This agent has been used for rRNA gene expression studies of Triticeae amphiploids and hybrids regarding rye rRNA genes suppression caused by the wheat nucleolar dominance phenomenon. However, this situation is reverted by 5-AC treatment which activates rye rRNA gene expression as it has been intensively observed in triticale. For nucleolar dominance studies, we produced F1 multigeneric hybrids (AABBRH^ch; 2n = 6x = 42) from crosses between the triticale cultivar ‘Corgo’ (AABBRR; 2n = 6x = 42) and the tritordeum cultivars HT9 and HT31 (AABBH^chH^ch; 2n = 6x = 42). The hybrid seeds were germinated in a low concentration of 5-AC (treatment) and in distilled water (nontreated control plants). Silver nitrate staining performed in one 5-AC-treated F1 hybrid revealed a reduced number of interphase cells with seven nucleoli, metaphases with eight Ag-NORs, and neocentromeres in the long arm of three wheat chromosomes. Nontreated hybrids presented six Ag-NORs per mitotic metaphase cell and a maximum of six nucleoli per interphase because of the 1R Ag-NOR suppression. No neocentromere was found in the control F1 hybrid plants. Both treated and nontreated seedlings were subsequently evaluated by fluorescent in situ hybridization performed with genomic and repetitive DNA probes to identify H^ch and rye genomes, to confirm Ag-NORs location, and to detect inactive rDNA loci. DAPI counterstaining was also helpful for the detection of neocentromeres in the long arm of three wheat chromosomes. This study allowed us to suggest that 5-AC treatment specifically induced wheat neocentromeres in the F1 multigeneric triticale × tritordeum hybrids.     

The influence of the rye genome on expression of heat shock proteins in triticale

Summary The heat shock protein profiles from Secale cereale L. cv Imperial, Triticum aestivum L. cv Chinese Spring, S. cereale x T. aestivum amphiploid, and the seven disomic S. cereale addition lines to T. aestivum were used to compare the wheat, rye, and triticale Heat Shock Protein profiles and to study the influence of the rye genome on heat shock protein expression in triticale. Three-day-old seedlings were heat shocked for 2 h at 40 °C in the presence of ³⁵S-methionine, and polypeptides from root tissues were subjected to one- or two-dimensional gel electrophoresis. The wheat and rye heat shock protein profiles each consisted of > 150 heat shock proteins, of which 94 were sufficiently reproducible to construct a standard map. There were 11 unique rye heat shock proteins compared to 22 unique wheat heat shock proteins. The triticale heat shock protein profile resembled the rye parent more than the wheat parent. There were 22 heat shock proteins expressed uniquely by wheat that were not expressed in triticale. Rye chromosomes 1 and 3 exhibited a substantial repressive influence on the expression of 95% of the unique wheat heat shock proteins in triticale, while rye chromosome 4 appeared to have the least repressive influence on expression of the unique wheat heat shock proteins in triticale.Mention of a trade name or proprietary product does not constitute a guarantee, warranty, or recommendation of the product by the United States Department of Agriculture or the University of Missouri and does not imply its approval to the exclusion of other products that may be suitable     

Development of commercially valuable traits in hexaploid triticale lines with Aegilops introgressions as dependent on the genome composition

Introgressive hybridization is an efficient means to improve the genetic diversity of cultivated cereals, including triticale. To identify the triticale lines with Aegilops introgressions, genotyping was carried out with ten lines obtained by crossing hexaploid triticale with genome-substitution forms of the common wheat cultivar Avrora: Avrolata (AABBUU), Avrodes (AABBSS), and Avrotika (AABBTT). The genome composition of the triticale lines was studied by in situ hybridization, and recombination events involving Aegilops and/or common wheat chromosomes were assumed for nine out of the ten lines. Translocations involving rye chromosomes were not observed. Substitutions for rye chromosomes were detected in two lines resulting from crosses with Avrolata. Genomic in situ hybridization (GISH) with Ae. umbellulata DNA and molecular genetic analysis showed that chromosome 1R was substituted with Ae. umbellulata chromosome 1U in one of the lines and that 2R(2U) substitution took place in the other line. Fluorescence in situ hybridization (FISH) with the Spelt 1 and pSc119.2 probes revealed a translocation from Ae. speltoides to the long arm of chromosome 1B in one of the two lines resulting from crosses with Avrodes and a translocation in the long arm of chromosome 7B in the other line. In addition, the pSc119.2 probe revealed chromosome 1B rearrangements in four lines resulting from crosses with Avrolata and in a line resulting from crosses with Avrotika. The lines were tested for main productivity parameters. A negative effect on all productivity parameters was demonstrated for Ae. umbellulata chromosome 2U. The overwinter survival in all of the lines was similar to or even higher than in the original triticale cultivars. A substantial increase in winter resistance as compared with the parental cultivar was observed for the line carrying the T7BS-7SL translocation. The line with the 1R(1U) chromosome substitution seemed promising for the baking properties of triticale.     

Conversion of a RAPD-generated PCR product,containing a novel dispersed repetitive element,into a fast and robust assay for the presence of rye chromatin in wheat

Bulk segregant analysis was used to obtain a random amplified polymorphic DNA (RAPD) marker specific for the rye chromosome arm of the 1BL.1RS translocation, which is common in many high-yielding bread wheat varieties. The RAPD-generated band was cloned and end-sequenced to allow the construction of a pair of oligonucleotide primers that PCR-amplify a DNA sequence only in the presence of rye chromatin. The amplified sequence shares a low level of homology to wheat and barley, as judged by the low strength of hybridization of the sequence to restriction digests of genomic DNA. Genetic analysis showed that the amplified sequence was present on every rye chromosome and not restricted to either the proximal or distal part of the 1RS arm. In situ hybridization studies using the amplified product as probe also showed that the sequence was dispersed throughout the rye genome, but that the copy number was greatly reduced, or the sequence was absent at both the centromere and the major sites of heterochromatin (telomere and nucleolar organizing region). The probe, using both Southern blot and in situ hybridization analyses, hybridized at a low level to wheat chromosomes, and no hybridizing restriction fragments could be located to individual wheat chromosomes from the restriction fragment length polymorphism (RFLP) profiles of wheat aneuploids. The disomic addition lines of rye chromosomes to wheat shared a similar RFLP profile to one another. The amplified sequence does not contain the RIS 1 sequence and therefore represents an as yet undescribed dispersed repetitive sequence. The specificity of the amplification primers is such that they will provide a useful tool for the rapid detection of rye chromatin in a wheat background. Additionally, the relatively low level of cross-hybridization to wheat chromatin should allow the sequence to be used to analyse the organization of rye euchromatin in interphase nuclei of wheat lines carrying chromosomes, chromosome segments or whole genomes derived from rye.     

Polyploidization as a retraction force in plant genome evolution: sequence rearrangements in triticale

Background

Polyploidization is a major evolutionary process in plants where hybridization and chromosome doubling induce enormous genomic stress and can generate genetic and epigenetic modifications. However, proper evaluation of DNA sequence restructuring events and the precise characterization of sequences involved are still sparse.

Methodology/Principal Findings

Inter Retrotransposons Amplified Polymorphism (IRAP), Retrotransposons Microsatellite Amplified Polymorphism (REMAP) and Inter Simple Sequence Repeat (ISSR) largely confirmed the absence of any intraspecific variation in wheat, rye and triticale. The comparative analysis of banding profiles between wheat and rye inbred lines revealed 34% of monomorphic (common to both parental species) bands for the ten different primer combinations used. The analysis of triticale plants uncovered nearly 51% of rearranged bands in the polyploid, being the majority of these modifications, due to the loss of rye bands (83%). Sequence analysis of rye fragments absent in triticale revealed for instance homology with hydroxyproline-rich glycoproteins (HRGP), a protein that belongs to a major family of inducible defence response proteins. Conversely, a wheat-specific band absent in triticale comprises a nested structure of copia-like retrotransposons elements, namely Claudia and Barbara. Sequencing of a polyploid-specific band (absent in both parents) revealed a microsatellite related sequence. Cytological studies using Fluorescent In Situ Hybridization (FISH) with REMAP products revealed a widespread distribution of retrotransposon and/or microsatellite flanking sequences on rye chromosomes, with a preferential accumulation in heterochromatic sub-telomeric domains.

Conclusions/Significance

Here, we used PCR-based molecular marker techniques involving retrotransposons and microsatellites to uncover polyploidization induced genetic restructuring in triticale. Sequence analysis of rearranged genomic fragments either from rye or wheat origin showed these to be retrotransposon-related as well as coding sequences. Further FISH analysis revealed possible chromosome hotspots for sequence rearrangements. The role of chromatin condensation on the origin of genomic rearrangements mediated by polyploidization in triticale is also discussed.     

Visualization of Secale cereale DNA in wheat germ plasm by fluorescent in situ hybridization

Homozygous wheat/rye (1BL/1RS or 1AS/ 1RL) translocation lines have significantly contributed to wheat production, and several other wheat/rye translocation lines show a potential promise against biotic and abiotic stresses. Detecting the presence of rye at the chromosome level is feasible by C-banding and isozyme protocols, but the diagnostic strength of genomic in situ hybridization for eventually analyzing smaller DNA introgressions has greater significance. As a first step we have applied the genomic in situ hybridization technique to detect rye chromosomes in a wheat background using germ plasm of agricultural significance. By this method rye contributions to the translocations 1BL/1RS, 1AL/1RS, 5AS/5RL and 6BS/6RL could be identified. Differential labelling has further enabled the detection of rye and Thinopyrum bessarabicum chromosomes in a trigeneric hybrid of Triticum aestivum/Th. bessarabicum//Secale cereale.     

Use of RFLPs to determine the chromosome composition of tetraploid triticale (A/B)(A/B)RR.

Tetraploid triticale, (A/B)(A/B)RR (2n = 28), is a botanical novelty, an amphiploid composed of a diploid rye and a 14 chromosome wheat genome made up of chromosomes of the A and B genomes of tetraploid wheat. Restriction fragment length polymorphism (RFLP) markers were used to elucidate the chromosome composition of the mixed wheat genome of 35 different tetraploid triticale lines. Of 128 possible A/B chromosome pair combinations, only 6 were found among these lines, with a prevalence of the 1A, 2A, 3B, 4B, 5B, 6B, and 7B karyotype. In most triticale lines stable wheat genomes made up of only homologous A or B genome chromosome pairs were identified, however, in some lines homoeologous chromosome pairs were found. In this paper we demonstrate that RFLPs can be used successfully as an alternative to C-banding for the identification of the chromosome composition of tetraploid triticale and discuss the possible selective advantage of specific chromosome composition.     

Alterations and Abnormal Mitosis of Wheat Chromosomes Induced by Wheat-Rye Monosomic Addition Lines

Background

Wheat-rye addition lines are an old topic. However, the alterations and abnormal mitotic behaviours of wheat chromosomes caused by wheat-rye monosomic addition lines are seldom reported.

Methodology/Principal Findings

Octoploid triticale was derived from common wheat T. aestivum L. ‘Mianyang11’×rye S. cereale L. ‘Kustro’ and some progeny were obtained by the controlled backcrossing of triticale with ‘Mianyang11’ followed by self-fertilization. Genomic in situ hybridization (GISH) using rye genomic DNA and fluorescence in situ hybridization (FISH) using repetitive sequences pAs1 and pSc119.2 as probes were used to analyze the mitotic chromosomes of these progeny. Strong pSc119.2 FISH signals could be observed at the telomeric regions of 3DS arms in ‘Mianyang11’. However, the pSc119.2 FISH signals were disappeared from the selfed progeny of 4R monosomic addition line and the changed 3D chromosomes could be transmitted to next generation stably. In one of the selfed progeny of 7R monosomic addition line, one 2D chromosome was broken and three 4A chromosomes were observed. In the selfed progeny of 6R monosomic addition line, structural variation and abnormal mitotic behaviour of 3D chromosome were detected. Additionally, 1A and 4B chromosomes were eliminated from some of the progeny of 6R monosomic addition line.

Conclusions/Significance

These results indicated that single rye chromosome added to wheat might cause alterations and abnormal mitotic behaviours of wheat chromosomes and it is possible that the stress caused by single alien chromosome might be one of the factors that induced karyotype alteration of wheat.     

Meiotic pairing in hybrids between tetraploid Triticale and related species: new elements concerning the chromosome constitution of tetraploid Triticale

Summary Two F5 strains of tetraploid triticale (2n= 4x=28), obtained from 6x triticaleX2 rye progenies, were crossed with diploid and tetraploid rye, some durum and bread wheats, and various 8x and 6x triticale lines. Meiosis in the different hybrid combinations was studied. The results showed that the haploid complement of these triticales consists of seven chromosomes from rye and seven chromosomes from wheat. High frequencies of PMCs showing trivalents were observed in hybrids involving the reference genotypes of wheat and triticale. These findings proved that several chromosomes from the wheat component have chromosome segments coming from two parental wheat chromosomes. The origin of these heterogeneous chromosomes probably lies in homoeologous pairing occurring at meiosis in the 6x triticaleX2x rye hybrids from which 4x triticale lines were isolated. A comparison among different hybrids combinations indicated that the involvement of D-genome chromosomes in homoeologous pairing is quite limited. In contrast, meiotic patterns in 4x triticale X 2x rye hybrids showed a quite high pairing frequency between some R chromosomes and their A and B homoeologues.     

DNA content of wheat monosomics at interphase estimated by flow cytometry

 Two complete, independently maintained sets of 21 monosomic wheat lines derived from cv. ‘Chinese Spring’ were analyzed for their DNA content at the G1 stage with flow cytometry. The DNA content of individual chromosomes was estimated by subtracting the value of a monosomic line from that of euploid wheat. Our data show that the estimated 2C DNA of individual wheat chromosomes in 21 monosomics at the G1 stage ranges from about 0.58 pg in chromosome 1D to approximately 1.12 pg in chromosome 3A. The A genome (2C=6.15 pg) seems to contain more DNA than the B (2C=6.09 pg) and D (2C=5.05 pg) genomes. Analysis of variance showed significant differences (α=0.01) in DNA content both among homoeologous groups and among genomes. Our estimates of interphase DNA content of wheat chromosomes from monosomic lines were poorly correlated to the chromosome sizes at metaphase (r=0.622, P≤0.01). This poor correlation might be due to differential coiling among chromosomes during cell division, possible bias of fluorochrome binding to heterochromatin, or heterogeneity among monosomic lines. Finally, flow cytometry may aid but cannot replace cytological checks in aneuploid maintenance. Received: 21 January 1997 / Accepted: 23 June 1997