Avena Species Research Articles

Investigations of barley stripe mosaic virus as a gene silencing vector in barley roots and in Brachypodium distachyon and oat.

BackgroundGene silencing vectors based on Barley stripe mosaic virus (BSMV) are used extensively in cereals to study gene function, but nearly all studies have been limited to genes expressed in leaves of barley and wheat. However since many important aspects of plant biology are based on root-expressed genes we wanted to explore the potential of BSMV for silencing genes in root tissues. Furthermore, the newly completed genome sequence of the emerging cereal model species Brachypodium distachyon as well as the increasing amount of EST sequence information available for oat (Avena species) have created a need for tools to study gene function in these species.ResultsHere we demonstrate the successful BSMV-mediated virus induced gene silencing (VIGS) of three different genes in barley roots, i.e. the barley homologues of the IPS1, PHR1, and PHO2 genes known to participate in Pi uptake and reallocation in Arabidopsis. Attempts to silence two other genes, the Pi transporter gene HvPht1;1 and the endo-β-1,4-glucanase gene HvCel1, in barley roots were unsuccessful, probably due to instability of the plant gene inserts in the viral vector. In B. distachyon leaves, significant silencing of the PHYTOENE DESATURASE (BdPDS) gene was obtained as shown by photobleaching as well as quantitative RT-PCR analysis. On the other hand, only very limited silencing of the oat AsPDS gene was observed in both hexaploid (A. sativa) and diploid (A. strigosa) oat. Finally, two modifications of the BSMV vector are presented, allowing ligation-free cloning of DNA fragments into the BSMV-γ component.ConclusionsOur results show that BSMV can be used as a vector for gene silencing in barley roots and in B. distachyon leaves and possibly roots, opening up possibilities for using VIGS to study cereal root biology and to exploit the wealth of genome information in the new cereal model plant B. distachyon. On the other hand, the silencing induced by BSMV in oat seemed too weak to be of practical use. The new BSMV vectors modified for ligation-free cloning will allow rapid insertion of plant gene fragments for future experiments.

Phylogenetic investigation of Avena diploid species and the maternal genome donor of Avena polyploids

AbstractWe investigated the species and genome relationships among 82 Avena accessions representing thirteen diploid species (A and C genomes), six tetraploids (AB and AC genomes) and four hexaploids (ACD genome) to infer the evolutionary pathways in Avena using the plastid matK gene and the trnL‐F region. The matK and trnL‐F sequences pointed to the A‐genome diploid species as maternal parents of the polyploid species. Furthermore, different A‐genome diploid species might have served as the A genome donor of several different polyploid species. The probable ancestor of most hexaploid species, of the AC‐genome tetraploids, and of the AB‐genome species A. agadiriana was A. wiestii (As genome). The likely donor of the other three AB‐ genome tetraploids was A. hirtula (also As genome) and A. damascena (Ad genome) appears to be the A genome donor of the hexaploid A. fatua. The A genome origin of A. fatua differs from that for the other hexaploid Avena species, which is different from the common assumption that the hexaploid species evolved from a single hexaploid ancestor followed by gain or loss of domestication genes. Thus, several separate maternal lineages might be involved in different polyploid species.

A Set of New Simple Sequence Repeat and Avenin DNA Markers Suitable for Mapping and Fingerprinting Studies in Oat (Avena spp.)

This study was conducted to develop and test new markers for use in molecular characterization of oat (Avena spp.). Results are presented for two polymerase chain reaction (PCR)‐based markers targeting multiple avenin protein loci and 16 new microsatellite or simple sequence repeat (SSR) markers. The markers were mapped in two segregating oat (Avena sativa L.) populations, ‘Kanota’ × ‘Ogle’ (K×O) and ‘Terra’ × ‘Marion’ (T×M), and were also tested across a diversity panel consisting of 35 cultivated varieties of oat and representatives of five other Avena species. All markers were robust, multiallelic, and highly polymorphic. The SSR markers, some highlighting multiple loci, mapped throughout the genome, and the avenin markers mapped to their expected locations in K×O and T×M. Also as expected, the avenin markers were shown to be linked to the stem rust resistance gene Pg9 in the population ‘OT328’ × ‘Dumont’ (OT×Du). Practical applications in variety identification and germplasm diagnostic assays were explored. The markers were able to distinguish closely related lines for fingerprinting purposes and were also useful for determining whether variation within a line was the result of residual heterozygosity or contamination. In addition, the markers were found to be useful for forensic purposes and for determining whether crossing and backcrossing had been successful.

Assessment of distribution and diversity of Avena sterilis L. and Avena fatua L. in cereal crops of Greece based on a 3-year survey and selected morphological traits

Under the view of species genetic erosion and the need of better crops, wild species could become important germplasm sources for breeding. At present, several wild Avena species are used as donors of valuable characters in oat breeding. In this study, wild oat distribution and diversity in wheat, barley and oat fields of Greece was examined by means of a 3-year survey in some typical cereal producing regions in the central part of the country. Greenhouse experiments were also conducted in order to compare seedlings grown from wild oat accessions of these regions under the same conditions. A total of 151 fields were surveyed and 158 accessions of wild oats were studied in terms of some morphological traits. The occurrence and distribution of A. sterilis was very high, while a noticeable and first-time analytically reported presence of A. fatua was evident in some of the surveyed wheat fields. The high diversity of wild oat was furthermore indicated by means of the pot experiments. The high degree of phenotypic variation in A. sterilis could be attributed to the earlier introduction of this species in the sampling area, compared to A. fatua. Wild oat distribution and diversity in a region seems crucial for the cereal production and is an important parameter which must be considered in evaluating efficacy of weed control strategies, while genes controlling these characters could be potentially used to improve cultivated forms of oats or other cereals.

Comparative cytogenetic analysis of Avena macrostachya and diploid C-genome Avena species

The chromosome set of Avena macrostachya Balansa ex Coss. et Durieu was analyzed using C-banding and fluorescence in situ hybridization with 5S and 18S-5.8S-26S rRNA gene probes, and the results were compared with the C-genome diploid Avena L. species. The location of major nucleolar organizer regions and 5S rDNA sites on different chromosomes confirmed the affiliation of A. macrostachya with the C-genome group. However, the symmetric karyotype, the absence of "diffuse heterochromatin" and the location of large C-band complexes in proximal chromosome regions pointed to an isolated position of A. macrostachya from other Avena species. Based on the distribution of rDNA loci on the C-genome chromosomes of diploid and polyploid Avena species, we propose a model of the chromosome alterations that occurred during the evolution of oat species.

Genomic and polyploid evolution in genus Avena as revealed by RFLPs of repeated DNA sequences

Phylogenetic relationships and genome affinities were investigated by utilizing all the biological Avena species consisting of 11 diploid species (15 accessions), 8 tetraploid species (9 accessions) and 4 hexaploid species (5 accessions). Genomic DNA regions of As120a, avenin, and globulin were amplified by PCR. A total of 130 polymorphic fragments were detected out of 156 fragments generated by digesting the PCR-amplified fragments with 11 restriction enzymes. The number of fragments generated by PCR-amplification followed by digestion with restriction enzymes was almost the same as those among the three repeated DNA sequences. A high level of genetic distance was detected between A. damascena (Ad) and A. canariensis (Ac) genomes, which reflected their different morphology and reproductive isolation. The A. longiglumis (Al) and A. prostrata (Ap) genomes were closely related to the As genome group. The AB genome species formed a cluster with the AsAs genome artificial autotetraploid and the As genome diploids indicating near-autotetraploid origin. The A. macrostachya is an outbreeding autotetraploid closely related with the C genome diploid and the AC genome tetraploid species. The differences of genetic distances estimated from the repeated DNA sequence divergence among the Avena species were consistent with genome divergences and it was possible to compare the genetic intra- and inter-ploidy relationships produced by RFLPs. These results suggested that the PCR-mediated analysis of repeated DNA polymorphism can be used as a tool to examine genomic relationships of polyploidy species.

RAPD and SSR analysis of wild oats (Avena species) from North West frontier Province of Pakistan

Genetic diversity of 10 biotypes of wild oats (Avena species) which differ in certain morphological and physiological characters were analyzed using 9 Randomly Amplified Polymorphic DNA (RAPD) and 2 Simple Sequence Repeat (SSR) primer sets. RAPD and SSR primers revealed different levels of genetic polymorphisms among the various biotypes. Range of genetic distances estimated by using individual primers was 0 – 100 % but on an average genetic distances were in the range of 30 - 65% indicating that these biotypes were genetically very diverse and possess a high degree of polymorphism. Key words: Wild oats, DNA, Polymorphism, RAPD, SSR.

Relationships among Avena species as revealed by consensus chloroplast simple sequence repeat (ccSSR) markers

Consensus chloroplast simple sequence repeat (ccSSR) makers were used to assess the genetic variation and genetic relationships of 80 accessions from 25 taxa of the genus Avena. Fifteen out of 16 ccSSR markers (93.75%) were polymorphic. A total of 51 alleles were detected at the 16 ccSSR loci. The number of alleles per locus ranged from 1 to 6, with an average of 3.2 alleles. Among these ccSSR loci, the highest polymorphism information content (PIC) value was 0.754, while the lowest PIC value was 0. The mean genetic similarity index among the 80 Avena accessions was 0.545, ranging from 0.188 to 1.000. To assess the usefulness of ccSSRs in separating and distinguishing between haplome (genome) groups, we used ordination by canonical discriminant analysis and classificatory discriminant analysis. Although discriminant analysis separated the haplome groups unequivocally, it was up to 69% predictive of correctly classifying an individual plant whose haplome(s) is unknown in the case where it belonged to the A haplome group, 75% where it belonged in the AC group, and almost 80% where it belonged in the ACD group. The analysis of genetic similarity showed that diploid species with the A haplome were more diverse than other species, and that the species with the As haplome were more divergent than other diploid species with the A haplome. Among the species with the C haplome, A. clauda was more diverse than A. eriantha and A. ventricosa. In the cluster analysis, we found that the Avena accessions with the same genomes and/or belonging to the same species had the tendency to cluster together. As for the maternal donors of polyploid species based on this maternally inherited marker, A. strigosa served as the maternal donor of some Avena polyploidy species such as A. sativa, A. sterilis and A. occidentalis from Morocco. A. fatua is genetically distinct from other hexaploid Avena species, and A. damascena might be the A genome donor of A. fatua. Avena lusitanica served as the maternal parents during the polyploid formation of the AACC tetraploids and some AACCDD hexaploids. These results suggested that different diploid species were the putative A haplome donors of the tetraploid and hexaploid species. The C genome species A. eriantha and A. ventricosa are largely differentiated from the Avena species containing the A, or B, or D haplomes, whereas A. clauda from different accessions were found to be scattered within different groups.

The origin of the C-genome and cytoplasm of Avena polyploids

The contribution of C-genome diploid species to the evolution of polyploid oats was studied using C-genome ITS-specific primers. SCAR analysis among Avena accessions confirmed the presence of C-genome ITS1-5.8S-ITS2 sequences in the genome of AACC and AACCDD polyploids. In situ hybridization and screening of more than a thousand rRNA clones in Avena polyploid species containing the C-genome revealed substantial C-genome rRNA sequence elimination. C-genome clones sequenced and Maximum Likelihood Parsimony analysis revealed close proximity to Avena ventricosa ITS1-5.8S-ITS2 sequences, providing strong evidence of the latter's active role in the evolution of tetraploid and hexaploid oats. In addition, cloning and sequencing of the chloroplastic trnL intron among the most representative Avena species verified the maternal origin of A-genome for the AACC interspecific hybrid formation, which was the genetic bridge for the establishment of cultivated hexaploid oats.

AFLP variation in 25 Avena species

Current molecular characterization of ex situ plant germplasm has placed more emphasis on cultivated gene pools and less on exotic gene pools representing wild relative species. This study attempted to characterize a selected set of germplasm accessions representing various Avena species with the hope to establish a reference set of exotic oat germplasm for oat breeding and research. The amplified fragment length polymorphism (AFLP) technique was applied to screen 163 accessions of 25 Avena species with diverse geographic origins. For each accession, 413 AFLP polymorphic bands detected by five AFLP primer pairs were scored. The frequencies of polymorphic bands ranged from 0.006 to 0.994 and averaged 0.468. Analysis of molecular variance revealed 59.5% of the total AFLP variation resided among 25 oat species, 45.9% among six assessed sections of the genus, 36.1% among three existing ploidy levels, and 50.8% among eight defined genome types. All the species were clustered together according to their ploidy levels. The C genome diploids appeared to be the most distinct, followed by the Ac genome diploid A. canariensis. The Ac genome seemed to be the oldest in all the A genomes, followed by the As, Al and Ad genomes. The AC genome tetraploids were more related to the ACD genome hexaploids than the AB genome tetraploids. Analysis of AFLP similarity suggested that the AC genome tetraploid A. maroccana was likely derived from the Cp genome diploid A. eriantha and the As genome diploid A. wiestii, and might be the progenitor of the ACD genome hexaploids. These AFLP patterns are significant for our understanding of the evolutionary pathways of Avena species and genomes, for establishing reference sets of exotic oat germplasm, and for exploring new exotic sources of genes for oat improvement.

Molecular diversity of the 5S rRNA gene and genomic relationships in the genus Avena (Poaceae: Aveneae)

The molecular diversity of the rDNA sequences (5S rDNA units) in 71 accessions from 26 taxa of Avena was evaluated. The analyses, based on 553 sequenced clones, indicated that there were 6 unit classes, named according to the haplomes (genomes) they putatively represent, namely the long A1, long B1, long M1, short C1, short D1, and short M1 unit classes. The long and short M1 unit classes were found in the tetraploid A. macrostachya, the only perennial species. The long M1 unit class was closely related to the short C1 unit class, while the short M1 unit class was closely related to the long A1 and long B1 unit classes. However, the short D1 unit class was more divergent from the other unit classes. There was only one unit class per haplome in Avena, whereas haplomes in the Triticeae often have two. Most of the sequences captured belonged to the long A1 unit class. Sequences identified as the long B1 unit class were found in the tetraploids A. abyssinica and A. vaviloviana and the diploids A. atlantica and A. longiglumis. The short C1 unit class was found in the diploid species carrying the C genome, i.e., A. clauda, A. eriantha, and A. ventricosa, and also in the diploid A. longiglumis, the tetraploids A. insularis and A. maroccana, and all the hexaploid species. The short D1 unit class was found in all the hexaploid species and two clones of A. clauda. It is noteworthy that in previous studies the B genome was found only in tetraploid species and the D genome only in hexaploid species. Unexpectedly, we found that various diploid Avena species contained the B1 and D1 units. The long B1 unit class was found in 3 accessions of the diploid A. atlantica (CN25849, CN25864, and CN25887) collected in Morocco and in 2 accessions of A. longiglumis (CIav9087 and CIav9089) collected in Algeria and Libya, respectively, whereas only 1 clone of A. clauda (CN21378) had the short D1 unit. Thus there might be a clue as to where to search for diploids carrying the B and D genomes. Avena longiglumis was found to be the most diverse species, possibly harboring the A, B, and C haplomes. The long M1 and short M1 are the unit classes typical of A. macrostachya. These results could explain the roles of A. clauda, A. longiglumis, and A. atlantica in the evolution of the genus Avena. Furthermore, one clone of the tetraploid A. murphyi was found to have sequences belonging to the short D1 unit class, which could indicate that A. murphyi might have been the progenitor of hexaploid oats and not, as postulated earlier, A. insularis. The evolution of Avena did not follow the molecular clock. The path inferred is that the C genome is more ancient than the A and B genomes and closer to the genome of A. macrostachya, the only existing perennial, which is presumed to be the most ancestral species in the genus.

Expansion of PCR‐based Marker Resources in Oat by Surveying Genome‐Derived SSR Markers from Barley and Wheat

Identifying polymerase chain reaction (PCR)‐based markers in crop genomes and amplifying them with specific primer pairs has provided convenient molecular markers for mapping projects. Oat (Avena sativa L.) lags behind other crops in the utilization of PCR‐based markers due to limited development of genomic and genetic resources in Avena species. We surveyed 356 genome‐derived simple sequence repeat (SSR) markers from wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.), chosen on the basis of even dispersal across different chromosomes, to search for an alternate method of expanding the PCR‐based marker pool in oat. Primer pairs for these SSR markers were tested for amplification and polymorphism between parental lines from Ogle1040/TAM‐O‐301 (OT) and Kanota/Ogle157 (KO) mapping populations. Eighty‐nine of 210 wheat primer pairs (42%) and 56 of 146 barley primer pairs (38%) successfully amplified sequences in oat. Forty‐five percent of the amplified markers, representing 19% of the total markers, showed polymorphism between parental lines of at least one mapping population. The polymorphism was primarily the presence or absence of a product band. Fifteen PCR products from 10 primer pairs were tested for reproducibility by amplifying each marker in the OT population. When assayed with the same PCR conditions used in the survey, the segregation ratio of 14 markers did not differ from the 1:1 ratio expected for a single locus. This study indicates that genomic SSR primer pairs from wheat and barley may be a good way to efficiently generate PCR‐based DNA markers for oat genetics research.

On evolutionary pathways of Avena species

This article presents literary review and results analysis of evaluation of representative set of oat accessions of all Avena L. species. Results of complex study of major morphological characters and utilization of the karyotype structure data confirmed by the results of RAPD and avenin spectrum analysis are presented. Relationships of genomes of different Avena species at each ploidy level are discussed. Two genomes form the base of all Avena species, namely the A and C genomes. Results of the evaluation of several characteristics of the oat species and their geographical distribution are analysed. Probable evolutionary pathway of Avena species are suggested. Most likely the centres of origin of genus Avena L. are determined.

Optimising sporulation and virulence in Drechslera avenacea

Studies were conducted on agar media to optimise sporulation of Drechslera avenacea, a fungal pathogen being evaluated as a biological control agent for Avena species (wild oats). Conidium production was affected by nutrition, pH, temperature and light conditions. Of the agar media tested, Czapek Dox agar (CZA) and half-strength oatmeal agar (½OMA) were the only media where sporulation occurred at all temperatures tested under a 12-h light:12-h dark photoperiod (L/D). The optimum temperature for conidium production was 20°C on ½OMA, whereas there was no optimum temperature on CZA. Under a 12-h near-ultraviolet (NUV):12-h dark photoperiod (NUV/D), similar numbers of conidia were produced on CZA at 6.66, 14.56, and 22.78 W m−2, whereas on ½OMA conidium production was the highest at 14.56 W m−2. When NUV/D and L/D conditions were compared, similar numbers of conidia where produced on CZA, whereas ½OMA conidium production was superior under the NUV/D photoperiod. Considerable variation in sporulation and degree of virulence of D. avenacea was detected among isolates from different geographic areas. The most virulent conidia were obtained on ½OMA at 20°C incubated under continuous illumination NUV light. Therefore, the most suitable conditions for conidium production of D. avenacea were growth for 1 week on ½OMA at 20°C under continuous NUV at an intensity of 14.56 W m−2. Under these conditions, 1.1×105 conidia mL−1 were produced which is the highest sporulation yet reported for any Drechslera spp., which are traditionally poor sporulators.

Cloning and characterization of resistance gene analogs from Avena species

Sequences analogous to plant resistance genes of the NBS-LRR class were cloned from the genomic DNA of 11 Avena species with different genomes and levels of ploidy. Three pairs of degenerate primers were used, based on conserved DNA sequence motifs belonging to the NBS domain, and 33 sequences were identified. These were subdivided into 7 classes depending on nucleotide sequence identity. Despite the high level of degeneracy, the primers behaved in a highly selective way; the majority of sequences from the different species obtained with every primer combination showed strong identity and were considered homologous. For most species, only one sequence of each class was identified in each genome, suggesting that duplicated sequences are fairly divergent. The strong identity among specific NBS sequences precludes any conclusions being made on the evolution of these species. The genomic organization of the RGA sequences was explored using those of A. strigosa as probes in Southern blots involving digested DNA from 15 Avena species. The hybridization patterns showed wide diversity both among sequences within a species and among species for each sequence. However, the dendrogram generated using the RFLPs showed relationships among species to be in good agreement with those previously established using other molecular markers.

Genomic Configuration of the Autotetraploid Oat Species Avena macrostachya Inferred from Comparative Analysis of ITS1 and ITS2 Sequences: on the Oat Karyotype Evolution during the Early Events of the Avena Species Divergence

To examine the genomic structure of Avena macrostachya, internal transcribed spacers, ITS1 and ITS2, as well as nuclear 5.8S tRNA genes from three oat species with AsAs karyotype (A. wiestii, A. hirtula, and A. atlantica), and those from A. longiglumis (AlAl), A. canariensis (AcAc), A. ventricosa (CvCv), A. pilosa, and A. clauda (CpCp) were sequenced. All species of the genus Avena examined represented a monophyletic group (bootstrap index = 98), within which two branches, i.e., species with A- and C-genomes, were distinguished (bootstrap indices = 100). The subject of our study, A. macrostachya, albeit belonging to the phylogenetic branch of C-genome oat species (karyotype with submetacentic and subacrocentric chromosomes), has preserved an isobrachyal karyotype, (i.e., that containing metacentric chromosomes), probably typical of the common Avena ancestor. It was suggested to classify the A. macrostachya genome as a specific form of C-genome, Cm-genome. Among the species from other genera studied, Arrhenatherum elatius was found to be the closest to Avena in ITS1 and ITS structure. Phylogenetic relationships between Avena and Helictotrichon remain intriguingly uncertain. The HPR389153 sequence from H. pratense genome was closest to the ITS1 sequences specific to the Avena A-genomes (p-distance = 0.0237), while the differences of this sequence from the ITS1 of A. macrostachya reached 0.1221. On the other hand, HAD389117 from H. adsurgens was close to the ITS1 specific to Avena C-genomes (p-distance = 0.0189), while its differences from the A-genome specific ITS1 sequences reached 0.1221. It seems likely that the appearance of highly polyploid (2n = 12-21x) species of H. pratense and H. adsurgens could be associated with interspecific hybridization involving Mediterranean oat species carrying A- and C-genomes. A hypothesis on the pathways of Avena chromosomes evolution during the early stages the oat species divergence is proposed.

Identification of indigenous pasture and the effect of time of harvesting and nitrogen fertilizer in the northwestern Ethiopian highlands

More than 80 natural pasture species: 55 grasses, seven legumes and 23 others, were identified in different grassland types and altitudes in the northwestern Ethiopian highlands. Andropogon, Avena, Eragrostis, Eleusine, Cynodon, Cyperus, Digitaria, Hyparrhenia, Pennisetum, Setaria, Trifolium and Medicago species were dominant. The dry matter yield (DMY) of natural pasture was significantly affected by the month of harvesting and the highest yield was in November. Application of nitrogen fertilizer significantly increased the DMY of natural pasture. Copyright © 2005 Whurr Publishers Ltd.

Isolation and identification of Triticeae chromosome 1 receptor-like kinase genes (Lrk10) from diploid, tetraploid, and hexaploid species of the genus Avena.

The DNA sequence of an extracellular (EXC) domain of an oat (Avena sativa L.) receptor-like kinase (ALrk10) gene was amplified from 23 accessions of 15 Avena species (6 diploid, 6 tetraploid, and 3 hexaploid). Primers were designed from one partial oat ALrk10 clone that had been used to map the gene in hexaploid oat to linkage groups syntenic to Triticeae chromosome 1 and 3. Cluster (phylogenetic) analyses showed that all of the oat DNA sequences amplified with these primers are orthologous to the wheat and barley sequences that are located on chromosome 1 of the Triticeae species. Triticeae chromosome 3 Lrk10 sequences were not amplified using these primers. Cluster analyses provided evidence for multiple copies at a locus. The analysis divided the ALrk EXC sequences into two groups, one of which included AA and AABB genome species and the other CC, AACC, and CCCC genome species. Both groups of sequences were found in hexaploid AACCDD genome species, but not in all accessions. The C genome group was divided into 3 subgroups: (i) the CC diploids and the perennial autotetraploid, Avena macrostachya (this supports other evidence for the presence of the C in this autotetraploid species); (ii) a sequence from Avena maroccana and Avena murphyi and several sequences from different accessions of A. sativa; and (iii) A. murphyi and sequences from A. sativa and Avena sterilis. This suggests a possible polyphyletic origin for A. sativa from the AACC progenitor tetraploids or an origin from a progenitor of the AACC tetraploids. The sequences of the A genome group were not as clearly divided into subgroups. Although a group of sequences from the accession 'SunII' and a sequence from line Pg3, are clearly different from the others, the A genome diploid sequences were interspersed with tetraploid and hexaploid sequences.

Molecular and morphological characterisation of the Heterodera avenae species complex (Tylenchida: Heteroderidae)

Abstract Species of the Heterodera avenae complex, including populations of H. arenaria, H. aucklandica, H. australis, H. avenae, H. filipjevi, H. mani, H. pratensis and H. ustinovi, obtained from different regions of the world were analysed with PCR-RFLP and sequencing of the ITS-rDNA, RAPD and light microscopy. Phylogenetic relationships between species and populations of the H. avenae complex as inferred from analyses of 70 sequences of the ITS region and of 237 RAPD markers revealed that the cereal cyst nematode H. avenae is a paraphyletic taxon. The taxonomic status of the Australian cereal cyst nematode H. australis based on sequences of the ITS-rDNA and RAPD data is confirmed. Morphometrical and ITS-rDNA sequence analyses revealed that the Chinese cereal cyst nematode is different from other H. avenae populations infecting cereals and is related to H. pratensis. Bidera riparia Kazachenko, 1993 is transferred to the genus Heterodera as H. riparia (Kazachenko, 1993) comb. n. As a consequence, H. riparia Subbotin, Sturhan, Waeyenberge & Moens, 1997 becomes a junior secondary homonym and is renamed as H. ripae nom. nov. Morphological, morphometrical characters and RFLP profiles for identification of the nine species presently placed in the H. avenae species complex are given.

Fluorescence in situ hybridization mapping of Avena sativa L. cv. SunII and its monosomic lines using cloned repetitive DNA sequences.

Fluorescent in situ hybridization (FISH) employing multiple probes was used with mitotic or meiotic chromosome spreads of Avena sativa L. cv. SunII and its monosomic lines to produce physical chromosome maps. The probes used were Avena strigosa pAs120a (which hybridizes exclusively to A-genome chromosomes), Avena murphyi pAm1 (which hybridizes exclusively to C-genome chromosomes), A. strigosa pAs121 (which hybridizes exclusively to A- and D-genome chromosomes), and the wheat rDNA probes pTa71 and pTa794. Simultaneous and sequential FISH employing two-by-two combinations of these probes allowed the unequivocal identification and genome assignation of all chromosomes. Ten pairs were found carrying intergenomic translocations: (i) between the A and C genomes (chromosome pair 5A); (ii) between the C and D genomes (pairs 1C, 2C, 4C, 10C, and 16C); and (iii) between the D and C genomes (pairs 9D, 11D, 13D, and 14D). The existence of a reciprocal intergenomic translocation (10C-14D) is also proposed. Comparing these results with those of other hexaploids, three intergenomic translocations (10C, 9D, and 14D) were found to be unique to A. sativa cv. SunII, supporting the view that 'SunII' is genetically distinct from other hexaploid Avena species and from cultivars of the A. sativa species. FISH mapping using meiotic and mitotic metaphases facilitated the genomic and chromosomal identification of the aneuploid chromosome in each monosomic line. Of the 18 analyzed, only 11 distinct monosomic lines were actually found, corresponding to 5 lines of the A genome, 2 lines of the C genome, and 4 lines of the D genome. The presence or absence of the 10C-14D interchange was also monitored in these lines.