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Genetic integrity of ex situ Genebank collections.

The genetic integrity of four accessions of the cross-pollinating species rye (Secale cereale L.) was investigated. Seeds available from the first and most recent regeneration cycles, multiplied 8, 12 (twice) or 14 times were fingerprinted using microsatellite markers. In all four accessions the allele numbers and frequencies changed after regeneration. Alleles present in the original seed sample were not detectable in the regenerated populations, whereas on the other hand, alleles were found in the recent seed sample, which were not observed in the investigated plants of the original one.

Alleles↗

The genetic diversity of components of rye hybrids.

The genetic variability between 5 open-pollinated varieties of rye (Secale cereale L) and between the components of rye hybrids was estimated using PCR-based marker analysis. The 22 maternal single crosses and 11 restorers were the components of hybrids investigated in the preofficial trials at the Plant Breeding and Acclimatization Institute in Radzików (PBAI), the Danko Breeding Co. Ltd (DBC) and in the Poznań Breeding Co. Ltd (PBC) during the 2001 growing season. The PCR system using semispecific primers targeting the intron-exon junction sequences of plant genes was applied for the evaluation of the genetic diversity of rye breeding materials. The genetic distances between varieties were relatively low. The coefficients of dissimilarity did not exceed 0,15. The highest average distance between maternal hybrids and restorers (0,28) was found in materials from PBAI, whereas in materials from PBC and DBC the average distances were lower and reached 0,21 and 0,20 respectively. The cluster analysis based on PCR data indicated that hybrid components that originated from different breeding centres exhibited different genetic characters.

Crops, Agricultural↗

[The effect of rye chromosomes on callus induction and regeneration in callus cultures of immature embryos of wheat-rye substitution lines, Triticum aestivum L. cultivar Saratovskaia 29/Secale cereale L. cultivar Onokhoiskaia].

The effect of individual rye chromosomes on the induction of callus and the character of its regenerating capacity was studied with cultured immature embryos of wheat-rye (Triticum aestivum L. cv. Saratovskaya 29-Secale cereale L. cv. Onokhoiskaya) substitution lines. The genotypic diversity of the substitution lines proved to significantly affect variation of parameters characterizing the major types of callus cultures, that is, frequencies of embryogenic calli, which are capable of shoot regeneration, and of morphogenic calli, which produce root structures. Functioning in the genotypic background of common wheat cultivar Saratovskaya, chromosomes 2R and 3R of rye cultivar Onokhoiskaya stimulated significantly the induction of embryogenic callus highly capable of shoot regeneration. Rye chromosome 2R present in place of chromosome 2D in the common wheat genome suppressed the induction of callus producing root structures. Rye chromosomes 1R and 6R suppressed the induction of embryogenic callus capable of shoot regeneration.

Chromosomes, Plant↗

[Mechanisms of meiotic restitution and their genetic regulation in wheat-rye polyhaploids].

Meiosis has been studied in partially fertile wheat-rye F1 hybrids yielded by crosses Triticum aestivum (Saratovskaya 29 variety) x Secale cereale L. (Onokhoiskaya variety) (4x = 28). Hybrid self-fertility proved to be caused by formation of restituted nuclei, which appear after equational segregation of univalent chromosome in AI and sister chromatid non-separation in AII of meiosis, as well as after AI blockage in three different ways. Both types of meiotic restitution were found in each hybrid plant. Expression of the "meiotic restitution" trait varied significantly in polyhaploids of the same genotype (ears of the same plants, anthers of the same ear, microsporocytes of the same anther). Chromatin condensation in prophase proved to be related to the division type and univalent segregation in AI. During reduction segregation of univalents in AI, sister chromatid cohesion and chromosome supercondensation remained unchanged. The results obtained suggest that in the remote hybrids with haploid karyotype of the parental origin (polyhaploids), the program of two-stage meiosis may be fundamentally transformed to ensure one instead of two divisions. We propose that meiotic restitution is a result of special genetic regulation of the kinetochore organization (both structural and functional) and chromatin condensation, i.e. of major meiotic mechanisms.

Haploidy↗

[Effects of accumulated temperature and planting density on tillering dynamics of forage rye].

Using equal design method, a field experiment was carried out at Wuqiao field station of China Agricultural University in Hebei Province to study the effects of different accumulated temperature before winter and planting density on tillering dynamics of forage rye (Secale cereale L.). The results showed that the tiller number per plant was affected by sowing date and seed density. In the prophase of forage rye, the tiller number per plant was mainly influenced by sowing date; while in the anaphase, it was mainly determined by seed density. Generally, earlier and fewer sowing norm could raise the tiller number in individual plant. In the prophase, metaphase, or anaphase of forage rye, the tiller number per unit area decreased with the postponing of sowing date, and increased with the increase of sowing norm. The sowing date and sowing norm had decisive effect on the number of effective stem per unit area. Suitable population structure was dependent on the increase of effective stem number per plant under the conditions of earlier sowing, and of effective stem number per unit area under the conditions of later sowing.

Regression Analysis↗

[Molecular cloning and sequence analyzing of cytoplasmic ribosomal protein gene OsRPS7 from rice (Oryza sativa)].

Using the cDNA of rye cytoplasmic ribosomal protein ScRPS7 as a query probe, a highly homologous rice genomic contig was obtained from Huada rice genome database. The full-length cDNA sequence of rice cytoplasmic ribosomal protein S7 was assembled by informatics based on the contig. Furthermore, with the two primers designed according to this assembled cDNA, the full-length cDNA of rice ribosomal protein was cloned by RT-PCR and named as OsRPS7. The cDNA was 919bp in length and contained a complete Open Reading Frame (ORF) of 576bp, encoding a protein of 192 amino acid residues. The deduced amino acids of OsRPS7 showed 88%,72% and 72% identity with those from Secale cereale, Arabidopsis thaliana and Brassica oleracea, respectively. The genome structure of OsRPS7 was analyzed, and its function was predicted in this paper.

Amino Acid Sequence↗

[Actinomycete complexes in the rhizosphere of winter rye on soddy podzolic soil].

The structure of actinomycete complexes in the rhizosphere of winter rye (Secale cereale L.) varieties originating from different ecosystems and geographical zones was studied in field experiments on soddy podzolic soil. In addition to streptomycetes, the complexes studied contained actinomycetes of the genera Micromonospora and Streptosporangium, represented at high occurrence and comparable abundance rates. The rhizosphere of most of the studied rye varieties was dominated by micromonosporas. The antifungal potential of mycelial prokaryotes associated with winter rye was assessed. The taxonomic and functional structure of actinomycete complexes was shown to be similar in rye varieties originating from the nonchernozem zone of Russia. However, the actinomycete complex proved to be much different in the rye variety that developed in the steppe zone of the lower Volga region.

Actinobacteria↗

[Molecular analysis of the triticale lines with different Vrn gene systems using microsatellite markers and hybridization in situ].

Hexaploid triticale (x Triticosecale Wittmack) lines were examined using molecular markers and the hybridization in situ technique. Triticale lines were generated based on wheat varieties differing by the Vrn gene systems and the earing times. Molecular analysis was performed using Xgwm and Xrms microsatellite markers with the known chromosomal localization in the common wheat Triticum aestivum, and rye Secale cereale genomes. Comparative molecular analysis of triticale lines and their parental forms showed that all lines contained A and B genomes of common wheat and also rye homeologous chromosomes. In the three lines the presence of D genome markers, mapped to the chromosomes 2D and 7D, was demonstrated. This was probably the consequence of the translocations of homeologous chromosomes from wheat genomes, which took part during the process of triticale formation. The data obtained by use of genomic in situ hybridization supported the data of molecular genetic analysis. In none of the lines wheat--rye translocations or recombinations were observed. These findings suggest that the change of the period between the seedling appearance and earing time in triticale lines compared to the initial wheat lines, resulted from the inhibitory effect of rye genome on wheat vernalization genes.

Chromosomes, Plant↗

Rye SCAR markers for male fertility restoration in the P cytoplasm are also applicable to marker-assisted selection in the C cytoplasm.

The study aimed at testing the usefulness of recently developed SCAR markers on rye (Secale cereale L.) chromosome 4R in hybrid breeding based on the C source of male sterility-inducing cytoplasm. Of 10 markers studied, 4 revealed polymorphisms between 2 inbred lines (544cms-C and Ot0-20) crossed to develop F2 and BC1 mapping populations. Analyses performed on 94 F2 and 93 BC1 plants allowed to extend a formerly constructed genetic map of chromosome arm 4RL. Three SCAR markers (SCP14M55, SCP15M55 and SCP16M58) were mapped in the vicinity of gene Rfc1, which restores male fertility in the C cytoplasm. The 3 tested SCAR markers proved to be effective in marker-assisted selection (MAS) for male fertility/sterility.

Breeding↗

[Effects of sodium nitroprusside on mitochondrial function of rye and wheat root tip under aluminum stress].

Sodium nitroprusside (SNP) could ameliorate the inhibition effect of Al on root growth of rye (Secale cereale L. cv. King) and wheat (Triticum aestivum L. cv. Jinmai47). Respiratory rate, P/O, OPR (oxygenated phosphate rate), R(3) (oxygen consumption rate with ADP and substrate present), R(4) (oxygen consumption rate with substrate) and RCR (respiratory control ratio, R(3)/R(4)) of root tips from rye and wheat decreased, as well as the activities of H(+)-ATPase, H(+)-PPase, Na(+)-K(+)-ATPase, Ca(2+)-ATPase and Mg(2+)-ATPase, but they increased with SNP treatment. It showed that mitochondrial respiratory functions of root tips from rye and wheat were damaged, phosphorylation was un-coupled by Al, but that of rye was less than that of wheat. Rye has high Al-resistance ability than wheat. SNP is one of donor of NO, it is suggested that NO can ameliorate remarkably respiratory dysfunction resulted from Al stress, so that NO can ameliorate the inhibition effect of Al on plant growth.

Aluminum↗

[Colonization of plant rhizosphere by actinomycetes of different genera].

The survival of environmental isolates of actinomycetes introduced with the seeds of agricultural plants in the root-free soil and in the rhizosphere and rhizoplane was studied. Different strategies of colonization of the rhizosphere were revealed for the representatives of the genera Streptomyces, Micromonospora, and Streptosporangium, the organisms typical for the moderate climate rhizosphere. The plants of winter rye (Secale cereale L.) inoculated with actinomycetes were shown to have growth advantages, while the cow clover plants (Trifolium pratense L.) had no growth advantages compared to uninoculated plants. The role of the plant component in the interaction with mycelial prokaryotes is discussed.

Actinobacteria↗

[Production of wheat-rye substitution lines and identification of chromosome composition of karyotypes using C-banding, GISH, and SSR markers].

Based on the cross (Triticum aestivum L. x Secale cereale L.) x T. aestivum L., wheat-rye substitution lines (2n = 42) were produced with karyotypes containing, instead of a pair of homologous wheat chromosomes, a homeologous pair of rye chromosomes. The chromosome composition of these lines was described by GISH and C-banding methods, and SSR analysis. The results of genomic in situ hybridization demonstrated that karyotype of these lines included one pair of rye chromosomes each and lacked wheat--rye translocations. C-banding and SSR markers were used to identify rye chromosomes and determine the wheat chromosomes at which the substitution occurred. The lines were designated 1R(1D), 2R(2D)2, 2R(2D)3, 3R(3B), 6R(6A)2. The chromosome composition of lines IR(1A), 2R(W)1, 5R(W), 5R(5A), and 6R(W)1, which were earlier obtained according to the same scheme for crossing, was characterized using methods of telocentric analysis, GISH, C-banding, and SSR analysis. These lines were identified as 1R(1A), 2R(2D)1, 5R(5D), 5R(5A), and 6R(6A)1, C-banding of chromosomes belonging to line 1R(1A) revealed the presence of two translocated chromosomes (3DS.3DL-del. and 4AL.W) during simultaneous amplification of SSR markers located on 3DL and 4AS arms. The "combined" long arm of the newly derived chromosome 4A is assumed to be formed from the long arm of chromosome 4AS itself and a deleted segment 3DL. All examined lines are cytologically stable, except for 3R(3B), which does not affect the stability of rye 3R chromosome transfer. Chromosome identification and classification of the lines will permit them to be models for genetic studies that can be used thereafter as promising "secondary gene pools" for the purpose of plant breeding.

Chromosome Banding↗

[Number and structure of actinomycetes complexes in the rhizosphere winter rye, oat and red clover].

The actinomycetes complexes in the rhizosphere of three agricultural plants by using the methods of luminescense microscopy and cup sowing were investigated. It was established, that concentration of prokaryotic biomass and biomass of actinomycetes mycelium in rhizosphere of plants is higher than in free from the radicals to soil. Rhizosphera of the oat (Avena sativa L.) and red clover (Trifolium pratense L.) is colonized by Streptomyces, Micromonospora and olygospore species. Dominante actinomycetes of winter rye (Secale cereale L.) are classified into the genera Micromonospora. It was shown that numbers and biomass of actinomycetes mycelium were fond to decreased, diversity of actinomycetes in contrast is increased in the series: "winter rye--oat--red clover". In connection with ecological safety the capability of increase with prokaryotes naturally disease suppressive soil and stability of plants to pathogen is discussed.

Actinobacteria↗

Inhibition of adenine nucleotide transport in mitochondria of winter rye seedlings.

Inhibition of adenine nucleotide transport was studied in mitochondria of winter rye (Secale cereale) seedlings. It has been shown that plant mitochondria have a translocase similar to the adenine-nucleotide translocase of animal mitochondria, since atractyloside, at a concentration of 40 micro M, inhibits Chance's State 3 respiration 100%. It has been demonstrated that palmitoyl-CoA--formed by a mitochondrial system for activation of fatty acids--lowers the rate of State 3 respiration during succinate oxidation, if palmitate, CoA and ATP are introduced into the incubation medium simultaneously. This can be explained by an inhibition of transport of adenine nucleotides through the internal mitochondrial membrane. The inhibition in question is counteracted by addition of carnitine.

Atractyloside↗

Purification and some properties of a nuclease from rye germ nuclei.

1. An endonuclease has been isolated from the nuclei of rye (Secale cereale L) germ and partially purified. The enzyme shows optimum activity over the pH range 5.4-7.4 towards both DNA and RNA, and has no phosphomonoesterase or phosphodiesterase activity. 2. DNA is degraded by the rye germ nuclease to oligonucleotides of similar size, and RNA to oligonucleotides and mononucleotides containing a C-terminal 5'-phosphate group. 3. The rate of hydrolysis of nuclear acids by the enzyme decreases in the following order: native DNA greater than denatured DNA greater than RNA. Synthetic polynucleotides are hydrolysed at a rate decreasing in the order: poly(A) greater than poly(U) greater than poly(C) greater than poly(G).

Cell Nucleus↗

Alkaline ribonuclease from rye germ cytosol.

1. Alkaline ribonuclease (pH optimum 7.6) was isolated from rye (Secale cereale L) germ cytosol and partially purified; the preparation was devoid of other nucleolytic activities. 2. The enzyme is a typical endonuclease hydrolysing all phosphodiester bonds in RNA, yielding ultimately purine and pyrimidine nucleoside 2',3'-cyclic phosphates and the corresponding 3'-phosphates. Upon extensive digestion of synthetic polyribonucleotides, pyrimidine, but not purine, nucleoside 3'-phosphates are formed. The enzyme does not hydrolyse synthetic purine cyclic nucleotides. 3. The enzyme does not depolymerize double-stranded complexes of poly(A) and poly(U). 4. Susceptibility to photooxidation and inhibition by 2-hydroxy-5-nitrobenzyl bromide and N-bromosuccinimide implies the involvement of tryptophan residue in the active centre of the enzyme.

Binding Sites↗

[Pollen development in the anthers of several cereal strains and hybrids during cultivation in vitro].

In order to induce androgenesis in vitro anthers of some cereals were cultivated. The highest number of proembryos was obtained in the hybrid Triticale in F3 generation on Blayder's medium supplemented with 3 and 12% sucrose. Proembryos represented multi-nuclear and multicellular formations which stagnated at the globular stage of development. Origin of roots from calluses was not accompanied by formation of buds. There was no formation of embryos from pollen grains in case of lines Triticum aestivum and Secale cereale.

Edible Grain↗

Isolation and characterization of genome-specific DNA sequences in Triticeae species.

Two contrasting genome-specific DNA sequences were isolated from Aegilops speltoides (wild goat grass) and Hordeum chilense (wild barley), each representing more than 1% of the genomes. These repetitive DNA fragments were identified as being genome-specific before cloning by genomic Southern hybridization (using total genomic DNA as a probe), and hence extensive screening of clones was not required. For each fragment, up to six recombinant plasmid clones were screened and about half were genome-specific. Clone pAesKB52 from Ae. speltoides was a 763 bp EcoRI fragment, physically organized in simple tandem repeats and shown to localize to sub-telomeric chromosome regions of species with the Triticeae S-genome by in situ hybridization to chromosomes. The sequence data showed an internal duplication of some 280 bp, which presumably occurred before sequence amplification and dispersion, perhaps by unequal crossing-over or reciprocal translocation. In situ hybridization showed that the sequence distribution varied between closely related (S-genome) species. Clone pHcKB6 was a 339 bp DraI fragment from H. chilense, also tandemly repeated but more variable; loss of the DraI site resulting in a ladder pattern in Southern blots which had little background smear. In situ hybridization showed that the tandem repeats were present as small clusters dispersed along all chromosome arms except at a few discrete regions including the centromeres and telomeres. The clone hybridized essentially specifically to the H-genome of H. chilense and hence was able to identify the origin of chromosomes in a H. chilense x Secale africanum hybrid by in situ hybridization.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗