RESUMO
A new Phaeosphaeria sp. biotype was isolated from winter ryes in Poland during 1995. Two isolates, Sn23-1 and Sn48-1, were obtained from diseased leaves of cvs. Motto and Dankowskie, respectively. The rye Phaeosphaeria sp. represented by isolate Sn48-1 has similar pycnidiospore morphology and induces disease symptoms in cereals similar to Phaeosphaeria nodorum, the causal agent of Stagonospora nodorum blotch disease (4). The pathogen (Sn48-1) produces hyaline, cylindrical pycnidiospores that are mostly three-septate and measure 12.8 to 23.7 × 2.1 to 3.2 µm (average size = 16 × 2.6 µm) on water agar. A molecular comparison of several genes in isolates Sn23-1 and Sn48-1 revealed that the rye Phaeosphaeria sp. was different from P. nodorum. In the conserved alpha-box sequence (1,93 bp) of the MAT1-1 gene, a four nucleotide difference occurred between the wheat-biotype P. nodorum and isolates Sn23-1 and Sn48-1 (GenBank Accession Nos. AY072933 and AF322008). In addition, the length of the internal transcribed spacer (ITS) region of the nuclear rDNA was the same for the wheat-biotype P. nodorum and the two rye Phaeosphaeria sp. isolates. However, a six nucleotide discrepancy was found in the ITS region (GenBank Accession Nos. U77362 and AF321323). The beta-glucosidase (bgl1) and beta-tubulin (tubA) genes differ in length between the wheat-biotype P. nodorum and two rye Phaeosphaeria sp. isolates (2,3). The main difference was due to the intron sizes of these two genes. One extra nucleotide was found in the intron2 of the bgl1 gene (GenBank Accession Nos. AY683619 and AY683620) and the intron1 of the tubA gene (GenBank Accession Nos. AY786337 and AY786331), respectively, in these two rye Phaeosphaeria sp. isolates. Disease severity on the fifth leaf (GS15) of Polish wheat (Alba, Begra, and Liwilla), triticale (Bogo and Pinokio), and rye (Zduno) cultivars was assessed with one (resistant) to nine (susceptible) scales 14 days after inoculation. Aggressiveness of wheat-biotype P. nodorum isolate Sn26-1 and rye Phaeosphaeria sp. isolate Sn48-1 was significant (P < 0.01) in five cultivars except in the moderately resistant wheat cv. Liwilla. The rye Phaeosphaeria sp. isolate Sn48-1 severely affected Polish rye Zduno (8.3) and two triticale cultivars (6.5), while the infection by isolate Sn26-1 was moderate (3-4). On the contrary, the wheat-biotype P. nodorum isolate Sn26-1 was more aggressive on wheat (4.1 on moderately resistant Alba and 6.2 on highly susceptible Begra) than the rye Phaeosphaeria sp. isolate Sn48-1, which had a scale of 2.2 and 4.3, respectively. Under laboratory conditions, the rye isolate Sn48-1 was able to cross with the wheat-biotype P. nodorum isolate Sn26-1 that has an opposite mating-type (MAT1-2) gene, but few viable ascospores were produced (1). References: (1) P. C. Czembor and E. Arseniuk. Mycol. Res. 104:919, 2000. (2) A. Malkus et al. FEMS (Fed. Eur. Microbiol. Soc.) Lett. 249:49, 2005. (3) E. Reszka et al. Can. J. Bot. 83:1001, 2005. (4) M. J. Richardson and M. Noble. Plant Pathol. 19:159, 1970.
RESUMO
ABSTRACT Stagonospora nodorum blotch (SNB) often develops explosively on upper leaves and glumes of wheat. Inoculum for late season infections may arise from early disease foci in the lower canopy or from recent immigration of wind-dispersed ascospores. Research was conducted to determine if foci of SNB are present and secondary spread has occurred in fields before tiller elongation. We determined the incidence of infection by Stagonospora nodorum for plants sampled at the mid-tillering stage in 96 1-m(2) quadrats in each of two fields. Isolates of S. nodorum were recovered from 32 quadrats, one per infected plant where possible. Multilocus restriction fragment length polymorphism haplotypes were determined for each isolate. Of 55 isolates collected from one field, there were 22 distinct haplotypes. Diseased plants were aggregated in both fields; aggregates sometimes extended to adjacent quadrats. Plants within aggregates were often infected by the same haplotype, suggesting that secondary spread had occurred. Foci overlapped because some aggregates were infected by more than one haplotype. Our results show that genetically diverse populations of S. nodorum were already established in fields before canopy development and were comprised of sometimes overlapping foci undergoing clonal expansion.
RESUMO
A repetitive sequence designated WE35 was isolated from wheat genomic DNA. This sequence consists of a 320-bp repeat unit and represents approximately 0.002% of the total wheat DNA. It is unidirectionally distributed either continuously or discretely in the genome. Ladder-like banding patterns were observed in Southern blots when the wheat genomic DNA was restricted with endonuclease enzymes EcoRI, HincII, NciI, and NdeI, which is characteristic for tandemly organized sequences. Two DNA fragments in p451 were frequently associated with the WE35 repetitive unit in a majority of lambda wheat genomic clones. A 475-bp fragment homologous to the 5'-end long terminal repeat (LTR) of cereal retroelements was also found in some lambda wheat genomic clones containing the repetitive unit. Physical mapping by fluorescence in situ hybridization (FISH) indicated that one pair of wheat chromosomes could be specifically detected with the WE35 positive probe p551. WE35 can be considered a chromosome-specific repetitive sequence. This repetitive unit could be used as a molecular marker for genetic, phylogenetic, and evolutionary studies in the tribe Triticeae.
Assuntos
DNA de Plantas/genética , Genoma de Planta , Sequências Repetitivas de Ácido Nucleico , Triticum/genética , Sequência de Aminoácidos , Sequência de Bases , Southern Blotting , Mapeamento Cromossômico , DNA de Plantas/isolamento & purificação , Hibridização in Situ Fluorescente , Dados de Sequência Molecular , Alinhamento de Sequência , Análise de Sequência de DNA , Triticum/ultraestruturaRESUMO
A new Zeta class glutathione S-transferases (GST) gene, pGST, has been cloned from wheat for the first time by the differential display PCR (DD-PCR) method. The genomic sequence of pGST, TA-GSTZ1, contains nine exons that encode a polypeptide of 213 amino acids and eight introns. The deduced amino acid sequence of TA-GSTZ1 as well as the exon:intron placement are more similar to the GSTs of the Zeta class than to the two wheat GSTs reported earlier. The pGST cDNA gene product expressed in Escherichia coli and purified by affinity chromatography showed typical Zeta class GST and glutathione peroxidase activities. Sequence polymorphism in the 3' untranslated region (UTR) of TA-GSTZ1 gene in wheat has been discovered. In this study, an 89 bp sequence is present in the 3' UTR of TA-GSTZ1gene in 16 wheat cultivars but absent in the other five. Although the biological importance of this polymorphism is unknown, it can be useful as a genetic marker in wheat breeding.
Assuntos
Genes de Plantas , Glutationa Transferase/genética , Triticum/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Evolução Molecular , Humanos , Dados de Sequência Molecular , RatosRESUMO
Two partial 1-aminocyclopropane-1-carboxylic acid (ACC) synthase cDNA clones (pWAS1, 1089 bp; and pWAS3, 779 bp) were isolated by polymerase chain reaction (PCR) using cDNA to total mRNA purified from etiolated wheat seedlings as template and degenerate oligonucleotides synthesized based on the regions of the ACC synthase amino acid sequence that are highly conserved among different plants. Northern analysis showed that the expression of the corresponding genes are differentially regulated. While the transcripts of pWAS1 were found in all the tissues of wheat that were tested with a maximum level at the early stages of spike development, pWAS3 mRNA was present almost exclusively in the root. A 5590 bp genomic clone, TA-ACS2, corresponding to pWAS3 cDNA has been isolated. The TA-ACS2 sequence consists of a 589-bp 5'-upstream region, 2743 bp of transcribed region with four exons and three introns and a 3'-downstream region of 2257 bp. Expression in Escherichia coli confirmed the ACC synthase activity of TA-ACS2 polypeptide. Sequence comparisons show that the two wheat ACC synthases are more similar to each other and to the rice ACC synthase, OS-ACS1, at the nucleotide level than at the amino acid level. The amino acid sequence of TA-ACS2 is most similar (66.1% identity) to that of broccoli. The chromosomal location of both wheat ACC synthase genes have been determined by aneuploid analysis. TA-ACS1 is located on the short arm of chromosomes 7A and 7D and on the long arm of chromosome 4A. TA-ACS2 is located on the long arm of homoeologous group 2 chromosomes.
Assuntos
Genes de Plantas , Isoenzimas/genética , Liases/genética , Raízes de Plantas/enzimologia , Triticum/genética , Sequência de Aminoácidos , Sequência de Bases , Mapeamento Cromossômico , Clonagem Molecular , DNA Complementar/genética , Escherichia coli/genética , Expressão Gênica , Isoenzimas/biossíntese , Liases/biossíntese , Liases/classificação , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , RNA Mensageiro/análise , RNA de Plantas/análise , Proteínas Recombinantes/biossíntese , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Distribuição Tecidual , Triticum/enzimologiaRESUMO
The MAV-PS1 and P-PAV isolates of barley yellow dwarf virus (BYDV) are serologically related, but not identical. Both are transmitted by the aphid Macrosiphum avenae, but P-PAV is also transmitted by Rhopalosiphum padi. To evaluate the basis for these and other differences, overlapping clones from cDNA libraries representing the genome of each isolate were characterized by restriction enzyme digestion and by hybridization, and subsequently sequenced. Each genome has six positive strand open reading frames (ORFs) which are similar to those identified from a BYDV isolate from Australia (Vic-PAV). The greatest diversity between MAV-PS1 and P-PAV sequences was found in ORFs located in the 3' half of the respective genomes, in particular ORFs 5 and 6, suggesting that these regions of the genome may be involved in the properties that differentiate MAV-PS1 and P-PAV. Sequence comparisons between P-PAV and Vic-PAV showed a high degree of identity in that all ORFs showed greater than 90% amino acid similarity, except ORF6 which had only 69% similarity.
Assuntos
DNA Viral/química , Vírus de Plantas/genética , RNA Viral/química , Proteínas Virais/genética , Sequência de Aminoácidos , Sequência de Bases , Biblioteca Gênica , Hordeum/microbiologia , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Fases de Leitura Aberta , Vírus de Plantas/classificação , Mapeamento por Restrição , Proteínas Virais/químicaRESUMO
Barley yellow dwarf virus (BYDV) can be separated into two groups based on, among other criteria, serological relationships that are presumably governed by the viral capsid structure. Nucleotide sequences for the coding regions of coat proteins of approximately 22 K were identified for the MAV-PS1, P-PAV (group 1) and NY-RPV (group 2) isolates of BYDV. The MAV-PS1 and P-PAV coat protein sequences shared 71% deduced amino acid similarity whereas that of the NY-RPV isolate shared no more than 51% similarity with either the MAV-PS1 or the P-PAV sequence. Other comparisons showed that these and other BYDV coat protein sequences examined to date share a high degree of identity with those identified from other luteoviruses. Among luteovirus coat protein sequences in general, several highly conserved domains were identified whereas other domains differentiate MAV-PS1 and PAV isolates from NY-RPV and other luteoviruses. Sequence similarities and differences among BYDV coat proteins (approx. 22K) are consistent with the serological relationships exhibited by these viruses. Amino acid sequence comparisons between BYDV isolates that share common aphid vectors indicate that it is unlikely that these coat proteins are involved in aphid specificity.
Assuntos
Capsídeo/genética , Genes Virais , Vírus de Plantas/genética , Proteínas Estruturais Virais/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , DNA Viral/genética , Biblioteca Gênica , Hordeum , Dados de Sequência Molecular , Fases de Leitura Aberta , Plasmídeos , Homologia de Sequência do Ácido NucleicoRESUMO
The utilization and conversion of D-xylose, D-xylulose, L-arabinose, and xylitol by yeast strains have been investigated with the following results: (1) The majority of yeasts tested utilize D-xylose and produce polyols, ethanol, and organic acids. The type and amount of products formed varies with the yeast strains used. The most commonly detected product is xylitol. (2)The majority of yeasts tested utilize D-xylulose aerobically and fermentatively to produce ethanol, xylitol, D-arabitol, and organic acids. The type and amount of products varies depending upon the yeast strains used. (3) Xylitol is a poor carbon and energy source for most yeasts tested. Some yeast strains produce small amounts of ethanol from xylitol. (4) Most yeast strains utilize L-arabinose, and L-arabitol is the common product. Small amounts of ethanol are also produced by some yeast strains. (5) Of the four substrates examined, D-xylulose was the perferred substrate, followed by D-xylose, L-arabinose, and xylitol. (6) Mutant yeast strains that exhibit different metabolic product patterns can be induced and isolated from Candida sp. Saccharomyces cerevisiae, and other yeasts. These mutant strains can be used for ethanol production from D-xylose as well as for the study of metabolic regulation of pentose utilization in yeasts.
RESUMO
Four yeasts (Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida utilus, and Rhodotorula toruloides) were tested for their ability to grow and consume D-glucose, D-xylose, D-xylulose, and D-xylitol. Sequential utilization of substrates was observed when D-glucose as mixed with D-xylulose as the carbon source. Catabolite inhibition was tentatively concluded to be responsible for this regulatory mechanism. D-Glucose was also found to inhibit the utilization of D-xylose and D-xylitol in C. utilus and R. toruloides. D-Xylose, D-xylitol, and D-xylulose were consumed simultaneously by R. toruloides and C. utilus.
Assuntos
Ascomicetos/metabolismo , Candida/metabolismo , Fungos Mitospóricos/metabolismo , Monossacarídeos/metabolismo , Rhodotorula/metabolismo , Saccharomyces cerevisiae/metabolismo , Schizosaccharomyces/metabolismo , Transporte Biológico , Galactose/metabolismo , Glucose/metabolismo , Cinética , Especificidade da Espécie , Xilitol/metabolismo , Xilose/metabolismo , Xilulose/metabolismoRESUMO
A high-d-xylulose mixture (d-xylose-d-xylulose = 33:67) was prepared from the cold ethanol extract of preisomerized d-xylose solution (d-xylose-d-xylulose = 77:23). Fusarium oxysporum f. sp. lini and Aspergillus niger were demonstrated to preferentially utilize d-xylose in the mixture of d-xylose and d-xylulose. Chromatographically pure d-xylulose was thus obtained in 90% yield. A high-d-xylulose mixture was also incubated with Rhodotorula toruloides, Klebsiella pneumoniae, Candida utilis, or Mucor rouxii.d-Xylose and d-xylulose were simultaneously consumed. When borate was added to the mixture, a d-xylulose-borate complex was formed, and it could be used to protect d-xylulose from being utilized.