Genomic and genetic relationships among species of Leymus (Poaceae: Triticeae) inferred from 18S-26S ribosomal genes

The 18S-26S ribosomal genes in three closely related species of Leymus (Poaceae: Triticeae) were examined using fluorescence in situ hybridization (FISH) and restriction fragment length polymorphism (RFLP). Both approaches revealed a close relationship between L. arenarius (8x = 56, northern European) and L. racemosus (4x = 28, central Eurasian), whereas L. mollis (4x = 28, northern American/Pacific) was distinct. Each species had three homologous pairs of major rDNA loci: a1, a2, and a3 for L. arenarius; m1, m2, and m3 for L. mollis; and r1, r2, and r3 for L. racemosus. Leymus arenarius had in addition three minor loci, a4, a5, and a6. The major loci of L. arenarius and L. racemosus were identical, indicating that the former species could have originated from the latter, via interspecific hybridization and/or polyploidy. The rDNA-RFLPs further indicated relationships of these species to other species of Leymus (L. karellini, 8x = 56 and L. angustus, 12x = 84) and Psathyrostachys (P. fragilis, P. huashanica, P. juncea, and P. lanuginosa, which are all diploids). A phenogram constructed from 20 BamHI, EcoRI, and DraI rDNA fragments revealed closer relationship between the two genera, Leymus and Psathyrostachys, than that among species within a genus.     

Phylogenetic relationships in Leymus (Poaceae: Triticeae) revealed by the nuclear ribosomal internal transcribed spacer and chloroplast trnL-F sequences

Using the nuclear ribosomal internal transcribed spacer (ITS) sequences and the chloroplasttrnL-F sequence, phylogeneic analysis was performed on 57 accessions of species in the tribe Triticeae including 13 Leymus species (N(s)) with different ploidy levels and 40 diploid species from 18 genera. The ITS sequences revealed that ployploid Leymus has close phylogentic relationships with Psathyrostachys and an undefined genus in Triticeae. The trnL-F tree demonstrated close relationships between certain Leymus species and Psathyrostachys, and other Leymus species distributed in North America were far from Psathyrostachys. Based on these results, it is unlikely that the unknown genome in Leymus species originated from one of the sampled diploid species in the present study. The maternal donor of all the Leymus species with a natural distribution in Eurasia were N(s) genome. Furthermore, Elymus californicus should be transferred from the genus Elymus to Leymus.     

Phylogenetic Analysis of Leymus (Poaceae: Triticeae) Inferred from Nuclear rDNA ITS Sequences

To investigate the phylogenetic relationships of polyploid Leymus (Poaceae: Triticeae), sequences of the nuclear rDNA internal transcribed spacer region (ITS) were analyzed for 34 Leymus accessions representing 25 species, together with three Psathyrostachys species (Ns genome), two Pseudoroegneria (St genome) species, Lophopyrum elongatum (E(e) genome), and Thinopyrum bessarabicum (E(b) genome). The phylogenetic analyses (maximum likelihood and Bayesian inference) supported two major clades, one including 21 Leymus species and three Psathyrostachys species, the other with nine Leymus species and four diploid species. The ITS RNA secondary structure of the Leymus species was compared with that of their putative diploid donor. It is suggested that (1) the species from the same areas or neighboring geographic regions are closely related to each other; (2) L. coreanus, L. duthiei, L. duthiei var. longearistatus, and L. komarovii are closely related to other Leymus species, and it is reasonable to transfer these species from the genus Hystrix to Leymus; (3) the ITS sequences of Leymus are evolutionarily distinct; (4) the different Leymus species and different distribution of a species derived their Ns genome from different Psathyrostachys species; and (5) there is a close relationship among Leymus, Pseudoroegneria, Lophopyrum, and Thinopyrum, but it is difficult to presume that the St, E(e), and E(b) genome may be the Xm genome donor of the Leymus species.     

Isolation, characterization, and analysis of Leymus-specific DNA sequences.

Genomic Southern hybridization using labeled total genomic DNA of Leymus mollis as probe showed intense hybridization signals on all restriction enzyme digested DNA from five species of Leymus Hochst., and four species of Psathyrostachys Nevski. Experiments using the same L. mollis probe, but with unlabeled blocking DNA from Psathyrostachys, showed no hybridization at all. These two genera evidently had the same genomic content. Southern hybridization without blocking allowed identification of DNA fragments abundant in Leymus and Psathyrostachys. Fragments potentially specific to Leymus were cloned. Five repetitive DNA clones from L. mollis and L. arenarius were characterized: pLmIs1, pLmIs44, pLmIs51, pLmIs53, and pLaIs56. These clones hybridized to both Leymus and Psathyrostachys on Southern blots - no clone hybridized to only one of these genera. Both Southern blot and fluorescence in situ hybridization (FISH) experiments showed that all the clones contained dispersed repetitive sequences. They painted all and whole chromosomes uniformly except at centromeres, telomeres, and nucleolar organiser regions. Three of these clones, i.e., pLmIs1, pLmIs44, and pLmIs53, were essentially specific to Leymus and Psathyrostachys - little or no hybridization was detected in other genera such as Triticum, Hordeum, Thinopyrum, or Elymus. Sequence analysis further revealed that the clones were part of retroelements. In particular, the clone pLmIs44 produced hybridization profiles suitable for analysis of genetic relatedness among species. The present study shows that Leymus and Psathyrostachys share the same basic genome, Ns, and therefore provides strong evidence for combining these two genera.     

Genetic and genomic relationships in Leymus Hochst

Genetic and genomic relationships among three taxonomically related species of Leymus, northern European L. arenarius (octoploid, 2n = 56), northern American/Pacific L. mollis (tetraploid, 2n = 28) and central Eurasian L. racemosus (tetraploid, 2n = 28), were examined using molecular and cytogenetic methods. The amplified fragment length polymorphism (AFLP) analysis clearly differentiated Icelandic populations of L. arenarius from Alaskan populations of L. mollis. The former group is more genetically homogeneous than the latter. Leymus arenarius in Iceland has a common gene pool and a relatively recent origin. The Alaskan L. mollis, on the other hand, is probably a glacial survival that has accumulated high level of genetic variation and has differentiated into subspecies. Analysis of the 18S-26S ribosomal genes, by restriction fragment length polymorphism (RFLP) and fluorescence in situ hybridization (FISH), revealed a very close relationship between the octoploid northern European L. arenarius and the tetraploid Eurasian L. racemosus, such that the former could have originated from the latter, probably via interspecific hybridization. Leymus-specific DNA sequences were isolated and used for analyzing genetic relatedness among five Leymus species and four Psathyrostachys species. The RFLP analysis of retrotransposon sequence pLm44 and ribosomal clone pTa71 clearly revealed a close relationship between these two genera, i.e. higher variation was found within genera than between them. The results support the previous notion that Leymus is autopolyploid having all genomes being designated Ns as in Psathyrostachys, but a major taxonomic revision of this group would require analysis of more species.     

Identifying the genome of wood barley Hordelymus europaeus (Poaceae: Triticeae)

Wood barley, Hordelymus europaeus, was compared with other Triticeae species by Southern and fluorescence in situ hybridisation using total genomic DNA and repetitive sequences as probes. On Southern blots, the total genomic probe from H. europaeus hybridised strongly to DNA of its own species and to Leymus and Psathyrostachys, indicating the presence of Ns genome in H. europaeus. Furthermore, the total genomic probe from P. fragilis hybridised to DNA of H. europaeus as much as to all of the Psathyrostachys and Leymus species examined. Ns genome-specific DNA sequences isolated from L. mollis (pLmIs1, pLmIs44 and pLmIs53) hybridised essentially to H. europaeus and all of the species of Leymus and Psathyrostachys. Chromosomal localization of these clones on H. europaeus confirmed the presence of Ns genome-specific DNA on all chromosomes, indiscriminately. Under moderate hybridisation stringency the Ns genome-specific probes, together with repetitive sequences pTa71 and pAesKB7, produced species-specific RFLP banding profiles on Southern blots. A phenetic tree based on these profiles revealed a distinct Ns species cluster within the Triticeae, represented by Leymus and Psathyrostachys species. Hordelymus europaeus belonged to this Ns cluster. Chromosomal mapping of the 18S-25S and the 5S ribosomal genes, together with the repetitive sequence pLrTaiI, corroborated that H. europaeus was most probably related to Leymus, especially the European/Eurasian members of sect. Leymus. In an attempt to identify the genome of H. europaeus, different approaches were employed; the results clearly showed that wood barley had the Ns basic genome and nothing else.     

大赖草染色体组构成的生化分析

本文通过对不同来源大赖草及其可能供体物种的EST、GOT、SOD同工酶及HMW-GS的分析, 发现不同来源大赖草的同工酶及蛋白图谱呈一定差异,但其中为不同来源大赖草所共有的一些谱带, 同时也存在于具N染色体组物种的分析图谱中,而在具J染色体组的物种中却没有特异性分布,推断大赖草中含有N染色体组,但可能不含J染色体组。 Abstract:The zymogram phenotypes of Esterase(EST),Glutamate oxalocate transminase(GOT),Superoxide dismutase(SOD)and the banding patterns of high molecular weight glutenin subunits(HMW-GS) were determined for different Leymus racemosus accessions and their putative donor species,It was found that although the zymogram phenotypes and the HMW-GS banding patterns appeared polymorphism,common bands for different Lr accessions were found in the species with N genome,but not in those with J genome,suggesting that the Psathyrostachys species with N genome must be phylogenetically closer to Lr,while the Thinopyrum species with J genome are farther to it.     

Fluorescence in situ Hybridization and Southern Blot Analyses on Leymus racemosus and Related Species

Fluorescence in situ hybridization (FISH) was carried out in somatic cells of racemosus (lam.) Tzvel. and Thinopyrum junceum ( Savul. & Rayss) A. Love using Th. bessarabicum ( Savul. & Rayss) A. Love genomic DNA as probe. Fourteen pairs of chromosomes in L. racemosus gave positive signal, and only seven of fourteen pairs of chromosomes in Th. junceum showed signal. In FISH probed with PHv62, the chromosomes in Th. bessarabicum and L. racemosus hybridized with PHv62, whereas Th. Bessarabicum showed positive signal in four pairs of chromosomes, and the latter in thirteen pairs of chromosomes. No positive signal was observed in chromosomes of Psathyrostachys juncea (Fisch.) Nevski and Th. junceum. The results of Southern hybridization probed with PHv62 were similar to those of in situ hybridization. Twelve alien chromosome lines of T. aestivum-L, racemosus were detected by PHv62 probing. In most of the alien chromosome lines, PHv62 hybridized in fragment of Leymus chromosomes, except for the line containing chromosomes 5Lr # 1 and 1 1Lr # 1. It is inferred that Th. bessarabicum may involve in the formation of species. Non the less, significant changes have occurred during the evolution of Leymus genomes.     

Phylogeny and molecular evolution of the DMC1 gene in the polyploid genus Leymus (Triticeae: Poaceae) and its diploid relatives

The level and pattern of nucleotide variation in duplicate genes provide important information on the evolutionary history of polyploids and divergent processes between homoeologous loci within lineages. Leymus, a group of allopolyploid species with the NsXm genomes, is a perennial genus with a diverse array of morphology, ecology, and distribution in Triticeae. To estimate the phylogeny and molecular evolution of a single-copy DMC1 gene in Leymus and its diploid relatives,DMC1 homoeologous sequences were isolated from the sampled Leymus species and were analyzed with those from 30 diploid taxa representing 18 basic genomes in Triticeae. Sequence diversity patterns and genealogical analysis suggested that: (i) different Leymus species might derive their Ns genome from different Psathyrostachys species; (ii) Pseudoroegneria has contributed to the nuclear genome of some Leymus species, which might result from recurrent hybridization or incomplete lineage sorting; (iii) the Xm genome origin of Leymus could differ among species; (iv) rapid radiation and multiple origin might account for the rich diversity, numbers of species, and wide ecological adaptation of Leymus species; and (v) the DMC1 sequence diversity of the Ns genome in Leymus species was lower than that in the Psathyrostachys diploids, while the level of DMC1 sequence diversity in Leymus was higher than that in diploid Pseudoroegneria. Our results provide new insight on the evolutionary dynamics of duplicate DMC1 genes, polyploid speciation, and the phylogeny of Leymus species.     

赖草(Leymus secalinus(Georgi)Tzvel.)中一个染色体标记的克隆与鉴定

通过构建Cot-1 DNA文库以及利用荧光原位杂交技术,从赖草(Leymus secalinus(Georgi)Tzvel.)中克隆获得一个在染色体多个部位具有点状杂交信号的重复序列。进一步对该序列在赖草基因组进行克隆和分析表明该序列为一个具有90 bp的重复单元(p Ls-90),并在基因组中呈串联状排列。利用p Ls-90对赖草进行分子核型分析,结果表明:p Ls-90在染色体的着丝粒、近着丝粒、臂间以及近端粒区均有分布,而且在每对染色体上信号分布模式不尽相同,结合染色体臂比可以清楚地对赖草的14对(28条)染色体进行识别。p Ls-90不仅可作为赖草种质鉴定和利用中染色体识别的理想标记,也可作为研究小麦族不同物种染色体组进化的有力工具。     

Molecular cytogenetics of Leymus: Mapping the Ns genome-specific repetitive sequences

The objective of this paper is to summarize the work in my group on FISH (fluorescent in situ hybridization) mapping of Ns-specific repetitive DNA sequences fromLeymus and discuss the results in the context of classification based on the genome system currently used among Triticeae researchers. The key question here is whether the genome composition of a tetraploid Leymus species should be NsXm or NsNs (Ns₁Ns₂). Different types of Leymus-specific dispersed retroelement-like repeats have been isolated and characterized. Because the sequences occur in significantly high copy number in Leymus, based on strong hybridization signal in Southern blots, they are considered essentially specific to Leymus. They are also abundant in Psathyrostachys, the progenitor of Ns genome in Leymus. These dispersed repeats are found to distribute over the whole of all Leymus chromosomes, without any differentiation between chromosomes that have been suggested to be of different genomic origins, meaning that all genomes in Leymus are the same. GISH (genomic in situ hybridization) experiments on Leymus chromosomes using Psathyrostachys genomic DNA as probes further support the NsNs (Ns₁Ns₂) genome constitution for Leymus. The Xm genome of an unknown origin might have been there in the beginning of the allopolyploidization process, but the Ns genome-specific elements must have spread predominantly and rapidly across genomes, thus homogenizing the nuclear genomes of Leymus. I present here for the first time evidence that Ns-specific dispersed repeats can spread in a very short time, from Leymus over to wheat in Triticum × Leymus hybrids growing in artificial conditions.     

Genetic diversity and phylogeny in Hystrix (Poaceae, Triticeae) and related genera inferred from Giemsa C-banded karyotypes

The phylogenetic relationships of 15 taxa from Hystrix and the related genera Leymus (NsXm), Elymus (StH), Pseudoroegneria (St), Hordeum (H), Psathyrostachys (Ns), and Thinopyrum (E) were examined by using the Giemsa C-banded karyotype. The Hy. patula C-banding pattern was similar to those of Elymus species, whereas C-banding patterns of the other Hystrix species were similar to those of Leymus species. The results suggest high genetic diversity within Hystrix, and support treating Hy. patula as E. hystrix L., and transferring Hy. coreana, Hy. duthiei ssp. duthiei and Hy. duthiei ssp. longearistata to the genus Leymus. On comparing C-banding patterns of Elymus species with their diploid ancestors (Pseudoroegneria and Hordeum), there are indications that certain chromosomal re-arrangements had previously occurred in the St and H genomes. Furthermore, a comparison of the C-banding patterns of the Hystrix and Leymus species with the potential diploid progenitors (Psathyrostachys and Thinopyrum) suggests that Hy. coreana and some Leymus species are closely related to the Ns genome of Psathyrostachys, whereas Hy. duthiei ssp. duthiei, Hy. duthiei ssp. longearistata and some of the Leymus species have a close relationship with the E genome. The results suggest a multiple origin of the polyploid genera Hystrix and Leymus.     

大赖草总DNA转化小麦的分子证据

用来自大麦组的４个高度重复序列克隆了ｐＨｖ７１６１,ｐＨｖ７１７８９,ｐＨｖ７１９１、ｐＨｖ７２９３,经地高辛和同位素２种方法标记后作为探针,对新疆大赖草（供体）、春麦７６１（受体）以及用大赖草总ＤＮＡ通过花粉管通道转化成功的大穗转化株基因组在高度严谨条件下进行了分子杂交。结果表明,这４个探针可以探查出基因内一种具有主串产重复单位的散在重复序列。比较受共体和转化体的杂交图谱,发现在转化株中出现了     

Absence of the J genome in Leymus species (Poaceae: Triticeae): evidence from DNA hybridization and meiotic pairing.

To test the presence of a J genome in the type species of Leymus, L. arenarius, its total genomic DNA and that of tetraploids L. mollis, L. salinus ssp. salmonis, L. ambiguus, L. chinensis, L. secalinus, L. alaicus ssp. karataviensis, and L. innovatus were probed with the 277-bp insert of pLeUCD2, which can hybridize with the J, S, and P but not with the N, R, V, Q, I, T, and ABD genomes. The DNA probe hybridized with PalI- or TaqI-digested total DNAs from Thinopyrum elongatum (JeJe diploid) and T. elongatum x Psathyrostachys juncea (JeN hybrid), but not with those from L. arenarius (NNNNXXXX octoploid) and all tetraploid Leymus species (NNXX). Attempts to cross diploid Thinopyrum and tetraploid Leymus species yielded only one triploid hybrid, T. elongatum x L. salinus ssp. salmonis. Meiotic chromosome associations at metaphase I of pollen mother cells in the triploid hybrid averaged 19.69 univalents, 0.64 bivalents, and 0.01 trivalents per cell. Chromosome pairings in the tetraploid hybrids of L. mollis x L. salinus ssp. salmonis, and the reciprocal cross, indicate that L. mollis and L. salinus ssp. salmonis shae the same genomic constitution. Both the DNA probe and genome analysis results confirm the absence of the J genome in the seven additional Leymus species tested. Meiotic data indicated that tetraploid Leymus species could not have the genome formula N1N1N2N2; thus their genome formulas should remain as NNXX until the source of X is identified.     

Development of Wheat-alien Lines with Added Leymus racemosus Chromosomes and 6VS/6AL Translocation Chromosomes

By chromosome C-banding and bi-color fluorescence in situ hybridization (FISH) using digoxigenin-labelled total genomic DNA of Leymus racemosus (Lam.) Tzvel. and biotinylated total genomic DNA of Haynaldia villosa (L.) Schur as probes, three wheat-alien lines with L. racemosus Lr.7 addition and H.　villosa 6VS/6AL translocated chromosomes, and eight lines with L.　racemosus Lr.14 addition and H.villosa 6VS/6AL translocated chromosomes were respectively identified from DALr.7×T6VS/6AL (93G51-4×P64) and DALr.14×T6VS/6AL （94G15×P64）F2 or F3 hybrids. Fluorescein-isothiocyanate-conjugated avidin and rhodamine-conjugated sheep anti-digoxigenin Fab fragment were used in bi-color FISH detection. The chromosomes of L.racemosus and 6VS fragment of H.　villosa were simultaneously detected by their red and green fluorescence. Powdery mildew and scab resistance were also evaluated. The result showed that the obtained plants had high resistance to these two diseases. The potential usage of bi-color FISH in identifying chromatin of L.racemosus and H.villosa was discussed.     

赖草的种间和属间杂种胚胎培养研究

以小麦族赖草属植物的６个种：赖草（Ｌｅｙｍｕｓｓｅｃａｌｉｎｕｓ）、窄颖赖草（Ｌ．ａｎｇｕｓｔｕｓ）、单小穗赖草（Ｌ．ａｅｍｕｌａｎｓ）、多枝赖草（Ｌ．ｍｕｌｔｉｃａｕｌｉｓ）、大赖草（Ｌ．ｒａｃｅｍｏｓｕｓ）和天山赖草（Ｌ．ｔｉａｎｓｈａｎｉｃｕｓ）作试验材料,分别组配成５个种间杂交组合,并与新麦草属（Ｐｓａｔｈｙｒｏｓｔａｃｈｙｓ）、鹅观草属（Ｒｏｅｇｎｅｒｉａ）以及Ｔｈｉｎｏｐｙｒｕｍ属组配成９个属间杂交组合。应用胚胎培养技术,离体培养杂种胚胎,成功地拯救了赖草植物的种间和属间杂种。试验结果表明,杂种胚胎的胚龄、培养基的营养组成以及杂种基因型对离体胚胎的培养有明显影响。     

Phylogeny of Hystrix and related genera (Poaceae: Triticeae) based on nuclear rDNA ITS sequences

The taxonomic status of Hystrix and phylogenetic relationships among Hystrix and its related genera of Pseudoroegneria (St), Hordeum (H), Psathyrostachys (Ns), Elymus (StH), Leymus (NsXm), Thinopyrum bessarabicum (E(b)) and Lophopyrum elongatum (E(e)) were estimated from sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA. The type species of Hystrix, H. patula, clustered with species of Pseudoroegneria, Hordeum, Elymus, Th. bessarabicum and Lo. elongatum, while H. duthiei ssp. duthiei, H. duthiei ssp. longearistata, H. coreana and H. komarovii were grouped with Psathyrostachys and Leymus species. The results indicate that: (i) H. patula is distantly related to other species of Hystrix, but is closely related to Elymus species; (ii) H. duthiei ssp. duthiei, H. duthiei ssp. longearistata, H. coreana and H. komarovii have a close affinity with Psathyrostachys and Leymus species, and H. komarovii might contain the NsXm genome of Leymus; and (iii) the St, H and Ns genomes in Hystrix originate from Pseudoroegneria, Hordeum and Psathyrostachys, respectively, while the Xm in Hystrix and Leymus has a complex relationship with the E or St genomes. According to the genomic system of classification in Tiritceae, it is reasonable to treat Hystrix patula as Elymus hystrix L, and the other species of Hystrix as species of a section of Leymus, Leymus Sect. Hystrix.     

Phylogenetic relationships in Leymus (Triticeae; Poaceae): Evidence from chloroplast trnH-psbA and mitochondrial coxII intron sequences

Leymus Hochst. is a polyploid genus with a diverse array of morphology, cytology, ecology, and distribution in Triticeae. To investigate the phylogenetic relationships and maternal genome donor of polyploid Leymus, the chloroplast trnH-psbA region and mitochondrial coxII intron sequences of 33 Leymus taxa were analyzed with those of 36 diploid perennial species representing 19 basic genomes in Triticeae. The results showed that reticulate evolution occurred in Leymus species, with the cytoplasmic lineage of Leymus contributed by different progenitors. Interspecific relationships of Leymus were also elucidated on the basis of orthologous comparison. Our data suggested that: (i) due to incomplete lineage sorting and/or difference in the pattern of chloroplast and mitochondrial inheritance, the genealogical conflict between the two genealogical patterns suggest the contribution of Psathyrostachys Nevski, Agropyron J. Gaertn, Eremopyrum (Ledeb.) Jaub. & Spach, Pseudoroegneria (Nevski) Á. Löve, Thinopyrum Á. Löve, and Lophopyrum (Host) Á. Löve to the cytoplasmic lineage of Leymus; (ii) there is a close relationship among Leymus species from the same area or neighboring geographic regions; (iii) L. coreanus (Honda) K. B. Jensen & R. R.-C. Wang, L. duthiei (Stapf) Y. H. Zhou & H. Q. Zhang ex C. Yen, J. L. Yang & B. R. Baum, L. duthiei var. longearistatus (Hack.) Y. H. Zhou & H. Q. Zhang ex C. Yen, J. L. Yang & B. R. Baum, and L. komarovii(Roshev.) C. Yen, J. L. Yang & B. R. Baum are closely related to other Leymus species, and it is reasonable to transfer these species from the genus Hystrix Moench to Leymus; (iv) Leymus species from North America are closely related to L. coreanus from the Russian Far East and L. komarovii from northeast China but are evolutionarily distinct from Leymus species from Central Asia and the Qinghai–Tibet Plateau. The occurrence of multiple origin and introgression could account for the rich diversity and ecological adaptation of Leymus species.     

Chromosomal localization of two novel repetitive sequences isolated from the Chenopodium quinoa Willd. genome

The chromosomal organization of two novel repetitive DNA sequences isolated from the Chenopodium quinoa Willd. genome was analyzed across the genomes of selected Chenopodium species. Fluorescence in situ hybridization (FISH) analysis with the repetitive DNA clone 18-24J in the closely related allotetraploids C. quinoa and Chenopodium berlandieri Moq. (2n = 4x = 36) evidenced hybridization signals that were mainly present on 18 chromosomes; however, in the allohexaploid Chenopodium album L. (2n = 6x = 54), cross-hybridization was observed on all of the chromosomes. In situ hybridization with rRNA gene probes indicated that during the evolution of polyploidy, the chenopods lost some of their rDNA loci. Reprobing with rDNA indicated that in the subgenome labeled with 18-24J, one 35S rRNA locus and at least half of the 5S rDNA loci were present. A second analyzed sequence, 12-13P, localized exclusively in pericentromeric regions of each chromosome of C. quinoa and related species. The intensity of the FISH signals differed considerably among chromosomes. The pattern observed on C. quinoa chromosomes after FISH with 12-13P was very similar to GISH results, suggesting that the 12-13P sequence constitutes a major part of the repetitive DNA of C. quinoa.     

Development of Triticum aestivum Leymus racemosus Translocation Lines by Irradiating Adult Plants at Meiosis

Two alien translocation lines referring to chromosome Lr. 7 of Leymus racemosus (lam.) Tzvel. were produced from M2 of Triticum aestivum-L, racemosus Lr. 7 monosomic addition lines irradiated with 60Co- γ rays of 600-1125R does shortly before meiosis. Among them, T02 was identified to contain largely-alien, partly-wheat translocation chromosomes, T08 contains a centric fusion translocation chromosome of TLr. 7S (L?) ·4AL on the basis of Giemasa C-banding and fluorescent in situ hybridization (FISH).