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Biological protection of the main cereals against fungal specific diseases.

Our field experiments showed that the use of Czech biopreparations (Supresivit, Ibefungin and Polyversum) based on the following microorganisms: Trichoderma harzianum, Bacillus subtilts and Pythium oligandrum applied as the seed treatment, the spray on the plants and like the mixture with mineral fertilizers (NPK, ammonium sulphate) lead namely after the seed-treatment and after the application as the mixture with mineral fertilizers to the increasing of the yield about 3-5 % (spring barley, winter wheat). This increasing was given by depression of soil-borne phytopathogenic fungi of the genera: Fusarium, Drechslera (Helminthosporium), Pseudocercosporella (Tapesia), Gaeumannomyces, and partially Rhynchosporium. The number of the fungi on plant rests were also influenced. No effect was observed on smuts and rusts. The smuts needed the seed-treatment with Vitavax (carboxim, thiram, imazalil) which didn't influence the biological treatment with biopreparations. The doses of biopreparations were following: the mixture with mineral fertilizer - 0.1 g of biopreparation/ kg of mineral fertilizer, the seed treatment - 0.1 g of biopreparation/1 kg of the seed and in the spray 0.1 g of biopreparation/1 litre of distilled water. Biological biopreparations will be usefull probably also in the future because they don't create the harmfull residues. The impact of the biopreparations under different ways of soil tillage (conventional variant with ploughing, variants with different plant residues and variants without ploughing) was studied as well. The influence of different soil preparation on composition of the soil mycoflora and the influence on quantity of the pathogenic genera Fusarium, Drechslera and others was observed. Quantity of named genera was negatively influenced while yield and health status of the plants were influenced positively.

Agriculture↗

Alkyl- and alkenylresorcinols of wheat grains and their chemotaxonomic significance.

Resorcinolic lipid contents and homologue compositions in extracts isolated from soft winter, soft spring and hard (durum) wheat grains were evaluated by both instrumental and chromatography means. Resorcinol concentrations determined in wheat were diverse and varied in samples harvested within two consecutive vegetative years, whereas their homologue profiles were found to be rather invariable. The predominant alkylresorcinols identified in wheat grains were saturated 1,3-dihydroxy-5-n-heneicosylbenzene and 1,3-dihydroxy-5-n-nonadecylbenzene. 1,3-Dihydroxy-5-n-heptadecylbenzene and 1,3-dihydroxy-5-n-tricosylbenzene were also determined, whereas 1,3-dihydroxy-5-n-pentadecylbenzene and 1,3-dihydroxy-5-n-pentacosylbenzene were present in these extracts only in spurious amounts. Furthermore, our results show that alk(en)ylresorcinols may be useful as chemotaxonomic markers for a distinction between soft and hard wheat plants. Cluster analysis with Ward's amalgamation algorithm and five different distance linkage types clearly discriminated particular wheats into species- and cultivar-specific clusters, whereas the use of principal component analysis allowed us to specify, which of the variables analysed were decisive. This approach may be useful for both plant breeders and taxonomists to classify wheat species/cultivars.

Phylogeny↗

Powdery mildew resistance genes in wheat: verification of STS markers.

Five accessions of Aegilops speltoides and 67 European wheat cultivars (winter and spring) originating from the Czech Republic, Germany, Poland, Russia, Slovakia, United Kingdom, and 4 non-European wheat cultivars from Brazil and the USA were examined with molecular Sequence Tagged Site (STS) markers for resistance genes to powdery mildew: Pm 1, Pm 2, Pm 3 and Pm 13. All markers gave clear, repeatable results, although three of them (Pm 1, Pm 2 and Pm 3) appeared as not specific for resistance genes. Comparison of STS analysis results with Pm genes, postulated as the reaction type after inoculation with differential isolates of Erysiphe graminis f.sp. tritici (Blumeria graminis), revealed a high number of disparities. The marker for Pm 13 was not detected in any examined cultivar but was present in five accessions of Aegilops speltoides.

Journal Article↗

Expression profile of two storage-protein gene families in hexaploid wheat revealed by large-scale analysis of expressed sequence tags.

To discern expression patterns of individual storage-protein genes in hexaploid wheat (Triticum aestivum cv Chinese Spring), we analyzed comprehensive expressed sequence tags (ESTs) of common wheat using a bioinformatics technique. The gene families for alpha/beta-gliadins and low molecular-weight glutenin subunit were selected from the EST database. The alignment of these genes enabled us to trace the single nucleotide polymorphism sites among both genes. The combinations of single nucleotide polymorphisms allowed us to assign haplotypes into their homoeologous chromosomes by allele-specific PCR. Phylogenetic analysis of these genes showed that both storage-protein gene families rapidly diverged after differentiation of the three genomes (A, B, and D). Expression patterns of these genes were estimated based on the frequencies of ESTs. The storage-protein genes were expressed only during seed development stages. The alpha/beta-gliadin genes exhibited two distinct expression patterns during the course of seed maturation: early expression and late expression. Although the early expression genes among the alpha/beta-gliadin and low molecular-weight glutenin subunit genes showed similar expression patterns, and both genes from the D genome were preferentially expressed rather than those from the A or B genome, substantial expression of two early expression genes from the A genome was observed. The phylogenetic relationships of the genes and their expression patterns were not correlated. These lines of evidence suggest that expression of the two storage-protein genes is independently regulated, and that the alpha/beta-gliadin genes possess novel regulation systems in addition to the prolamin box.

Base Sequence↗

Chromosomal Location of Genes Encoding Barley (1-->3, 1-->4)-beta-Glucan 4-Glucanohydrolases.

Preparations of DNA from wheat (Triticum aestivum, cv Chinese Spring), barley (Hordeum vulgare, cv Betzes) and six euplasmic wheat-barley addition lines were digested to completion with restriction endonucleases and the products probed by Southern blot analysis using a cDNA-encoding barley (1-->3, 1-->4)-beta-glucanase isoenzyme II. It is shown that one of the barley (1-->3, 1-->4)-beta-glucanase genes is located on chromosome 1.

Journal Article↗

QTL analysis of fertility-restoration against cytoplasmic male sterility in wheat.

The cytoplasm of Triticum timopheevi causes cytoplasmic male sterility (CMS) in common wheat (T. aestivum) cv. 'Chinese Spring' (CS), and that of Aegilops kotschyi causes CMS in spelt wheat (T. spelta) var. duhamelianum (Sp). CS has fertility-restoring (Rf) genes against the latter cytoplasm and Sp has the ones against the former. To know the genetic system concerning to CMS, we crossed 66 F8 recombinant inbred lines (RILs) derived from a cross between CS and Sp as males to the alloplasmic lines of CS and Sp having the cytoplasms of T. timopheevi and Ae. kotschyi, respectively. The fertilities of respective F1 plants derived from the crosses were examined for QTL analysis. The major QTLs detected in both systems were located on the short arm of chromosome 1B. One minor QTL on chromosome 2B was also commonly detected in both of the systems, while other minor QTLs against T timopheevi cytoplasm were distributed on the chromosomes 2A, 4B, and 6A.

Animals↗

Genetics and evolution of multilocus isozymes in hexaploid wheat.

Aneuploid genetic studies of isozyme variation in cv Chinese Spring have disclosed that numerous enzymes of hexaploid wheat exist in multiple molecular forms as a direct consequence of polyploidy. Sixty-nine isozyme structural genes have been identified to date. Two of these belong to a duplicate set and at least 54 to triplicate sets of paralogous genes that are located one each in related chromosomes in different genomes. Each of these gene sets encodes either two or three isozymes. The role of regional gene duplication in the production of multilocus isozymes in hexaploid wheat is as yet poorly understood, although a considerable amount of indirect evidence suggests that a large number of isozymes are encoded by genes that were produced by ancient regional gene duplication events in a genome ancestral to the genomes now present in the species. A full assessment of the role of regional gene duplication in the production of hexaploid wheat isozymes must await further studies. The isozyme structural gene locations thus far determined indicate that the gene synteny relationships that existed in the ancestral wheat genome are in large part conserved in each of the three genomes of cv Chinese Spring and that the genetic content of most individual chromosome arms has also been in large part conserved.

Biological Evolution↗

Single-strand-preferring nuclease activity in wheat leaves is increased in senescence and is negatively photoregulated.

Single-strand-preferring nucleases (EC 3.1.30.1) selectively cleave internucleotide bonds in single-stranded regions of predominantly duplex DNA and DNA.RNA hybrids and extensively degrade denatured DNA and RNA. The functions of single-strand-preferring nuclease in plants are unknown. We have monitored this nuclease activity in flag leaves of wheat (Triticum aestivum L. cv. Chinese Spring) undergoing natural senescence and in primary leaves of wheat seedlings undergoing dark-induced senescence. In falg leaves, nuclease activity remained at basal levels during the first 2 weeks after anthesis, while chlorophyll content increased to a maximum. Nuclease activity then rose in concert with a decline in chlorophyll, reaching a 16-fold elevation at 5 weeks post-anthesis, when 53% of the chlorophyll had been lost. When 8-day-old wheat seedlings were induced to senesce by placing them in darkness, nuclease activity rose without apparent lag, reaching a 13-fold elevation in 7 days, when 61% of the chlorophyll had been lost. The increase in nuclease activity was reversible upon reexposure of seedlings to light, a decline beginning without apparent lag. Reversibility was complete for plants that had been held in darkness for 5 days, with activity returning to the control level in 2 days. These senescence-related changes in nuclease activity, measured in conventional assays, were consistent with concomitant analysis by activity staining of sodium dodecyl sulfate/polyacrylamide gels. We conclude that an increase in single-strand-preferring nuclease activity is closely associated with wheat leaf senescence and that nuclease activity is subject to negative photoregulation.

Journal Article↗

Aneuploid analyses of three chlorophyll abnormalities in Emmer wheat.

Of the various chlorophyll abnormalities that occur in polyploid wheats, genetic bases of only two types, chlorina and virescence, are known. Here, for the first time, the chromosomal bases of three other chlorophyll abnormalities, striato-virescence, delayed virescence, and albino, which occur in Emmer wheat (2n = 4x = 28, genome constitution AABB) are reported. A set of disomic substitution lines of Langdon durum was used for chromosome identification. All three abnormalities are controlled by two duplicated recessive genes (carrier chromosome in parentheses); striato-virescence by sv1 (3A) and sv2 (2A), delayed virescence by dv1 (2B) and dv2 (supposedly 2A), and albino by abn1 (2A) and abn2 (2B). Genes on the same chromosome are located in different loci and are recombined with each other. The Chinese Spring cultivar of common wheat (2n = 6x = 42, genome constitution AABBDD) carries wild-type homoeoalleles for these abnormalities; Sv3 (2D), Dv3 (2D), and Abn3 (2D).

Aneuploidy↗

Fluorescent in situ hybridization and C-banding analyses of highly repetitive DNA sequences in the heterochromatin of rye (Secale montanum Guss.) and wheat incorporating S. montanum chromosome segments.

The molecular characterization of C-banded regions of Secale montanum Guss. by means of in situ hybridization was performed in order to provide new information about their chromosome structure relative to cultivated rye, Secale cereale L. Accurate identification of individual chromosomes was achieved using simultaneous and (or) successive fluorescent in situ hybridization (FISH) and C-banding. FISH identification was performed using total rye DNA, three highly repetitive rye DNA sequences (pSc119.2, pSc74, and pSc34), and the ribosomal RNA probes pTa71 (18S, 5.8S, and 26S rDNA) and pTa794 (5S rDNA). FISH was also used to identify the chromosome segment involved in two spontaneous translocation lines recovered from a 'Chinese Spring'--S. montanum wheat-rye addition line. FISH analysis revealed the exact translocation breakpoints and allowed the identification of the transferred rye segments. The value of this type of analysis is discussed.

Chromosome Banding↗

Variation in transformation frequencies among six common wheat cultivars through particle bombardment of scutellar tissues.

The transformation technique in common wheat has already been established by using microprojectile bombardment and scutellar tissues of immature embryos. In this study, in vitro culture response of immature embryos and the production of transgenic wheat plants were examined in six common wheat cultivars, i.e., Chinese Spring, Akadaruma, Haruhikari, Shiroganekomugi, Norin 12, and Norin 61. In all genotypes, more than seven hundred immature embryos were bombarded with a plasmid containing a bialaphos-resistant gene under control of the rice actin 1 gene. (Act1) promoter. Although the transient expression of the reporter gene encoding beta-glucuronidase following the rice Act1 promoter was similar in five of the six cultivars tested, the frequency of stable transformation varied with the genotype. The frequency of transformation was the highest in Akadaruma and Norin 12 of the six wheat cultivars; independently transformed plants were produced from 1.4% and 1.7% of bombarded embryos, respectively. On the other hand, the immature embryos of Norin 61 and Shiroganekomugi showing low efficiency of in vitro culture generated no transgenic plants. This variation of the transformation frequency was generally caused by the difference in the in vitro culture response with the genotype, rather than the efficiency of the introduction of the transgene into wheat cells by particle bombardment.

Fertility↗

Multiplex PCR identification of wheat HMW glutenin subunit genes by allele-specific markers.

Wheat bread-making quality is closely correlated with composition and quantity of gluten proteins, in particular with high-molecular weight (HMW) glutenin subunits encoded by the Glu-1 genes. A multiplex polymerase chain reaction (PCR) method was developed to identify the allele composition of HMW glutenin complex Glu-1 loci (Glu-A1, Glu-B1 and Glu-D1) in common wheat genotypes. The study of multiplex PCR to obtain a well-balanced set of amplicons involved examination of various combinations of selected primer sets and/or thermal cycling conditions. One to three simultaneously amplified DNA fragments of HMW glutenin Glu-1 genes were separated by agarose slab-gel electrophoresis and differences between Ax1, Ax2* and Axnull genes of Glu-A1 loci, Bx6, Bx7 and Bx17 of Glu-B1, and Dx2, Dx5 and Dy10 genes of Glu-D1 loci were revealed. A complete agreement was found in identification of HMW glutenin subunits by both multiplex PCR analysis and SDS-PAGE for seventy-six Polish cultivars/strains of both spring and winter common wheat. Rapid identification of molecular markers of Glu-1 alleles by multiplex PCR can be an efficient alternative to the standard separation procedure for early selection of useful wheat genotypes with good bread-making quality.

Alleles↗

[Study on chromosomes aberration in wheat-rye disomics addition lines induced by the gametocidal chromosome 2C].

In the present study,Chinese Spring-Imperial (1 R-7R) wheat-rye disomic addition lines were hybridized with Chinese Spring-2C (derived from Aegilops cylindrica) disomic addition lines. The F1 hybrids were examined by mitotic and meiotic analysis. There were observed abnormal chromosome configurations. A total of 430 F2 plants were obtained by self-pollination. Chromosomes aberrations, such as translocation, deletions, isobrachial and dicentromere chromosomes, are identified in F2 individual plants by C-banding combined with fluorescent in situ hybridization (FISH). Additionally, chromosome spontaneous substitutions such as 2C substituting for wheat chromosomes 2A, 2B and 2D were also observed. The rule and frequency of chromosome aberration in F2 are the following: 22 out of 430 F2 plants (5.11%) were found involving aberration rye chromosomes. Among them, 10 plants were identified as wheat-rye chromosome translocation lines comprising 2.3%. Rye chromosome deletions comprised 12 of them (2.79%). 3 isobrachial aberrations were detected (about 0.7%), too. Most of the translocation lines are with wheat centromere, only one of them is with rye centromere. Rye chromosome aberrations occurred unevenly among homoeologous groups. There were 5 in 1R, 3 in 2R, 1 in 3R, 3 in 4R, 6 in 5R and 4 in 6R. The majority of the translocation lines are terminal translocation. 54 out of the total 430 progenies are wheat deletions,and 27 are distributed in the A group, 20 in the B group and 7 in the D group respectively. Finally,we discussed the possible cause for the uneven chromosome aberration among homoeologous groups in wheat and rye as well as the effect characteristics of 2C on wheat and rye chromosome.

Chromosome Aberrations↗

Importance of insoluble-bound phenolics to antioxidant properties of wheat.

Two commercial samples of soft (70% Canadian Eastern soft red spring and 30% Canadian Eastern soft white winter) and hard (90% Canadian western hard red spring and 10% Canadian Eastern hard red winter) wheats were used to obtain different milling fractions. Phenolics extracted belonged to free, soluble esters and insoluble-bound fractions. Soluble esters of phenolics and insoluble-bound phenolics were extracted into diethyl ether after alkaline hydrolysis of samples. The content of phenolics was determined using Folin-Ciocalteu's reagent and expressed as ferulic acid equivalents (FAE). The antioxidant activity of phenolic fractions was evaluated using Trolox equivalent antioxidant capacity, 2,2-diphenyl-1-picrylhydrazyl radical scavenging, reducing power, oxygen radical absorbance capacity, inhibition of oxidation of human low-density lipoprotein cholesterol and DNA, Rancimat, inhibition of photochemilumenescence, and iron(II) chelation activity. The bound phenolic content in the bran fraction was 11.3 +/- 0.13 and 12.2 +/- 0.15 mg FAE/g defatted material for hard and soft wheats, respectively. The corresponding values for flour were 0.33 +/- 0.01 and 0.46 +/- 0.02 mg FAE/g defatted sample. The bound phenolic content of hard and soft whole wheats was 2.1 (+/-0.004 or +/-0.005) mg FAE/g defatted material. The free phenolic content ranged from 0.14 +/- 0.004 to 0.98 +/- 0.05 mg FAE/g defatted milling fractions of hard and soft wheats examined. The contribution of bound phenolics to the total phenolic content was significantly higher than that of free and esterified fractions. In wheat, phenolic compounds were concentrated mainly in the bran tissues. In the numerous in vitro antioxidant assays carried out, the bound phenolic fraction demonstrated a significantly higher antioxidant capacity than free and esterified phenolics. Thus, inclusion of bound phenolics in studies related to quantification and antioxidant activity evaluation of grains and cereals is essential.

Antioxidants↗

[Effect of plant residues on the parasitic activity of soilborne pathogens and the saprophytic microflora of the soil. III. Influence of rye and rape grown as winter catch crop on the incidence of Cercosporella herpotrichoides Fron (author's transl)].

In a five-years field trial, the influence of two winter catchcrops, rye and rape, upon the incidence of Cercosporella herpotrichoides has been studied. The winter catch-crops had been repeated three times, followed either by stubble-crop or without in fall of the fourth year. The variants (blocks) had been completed by two variants without catch-crop, either carefully cultivated or overgrown with weeds. Each of the four blocks had been divided three times to be grown with three different crop sequences containing 80%, 60% (with the insusceptible corn), or 40% haulm fruit, respectively. Each of the 12 variants had been divided into two parts by continuous fallow stripe, one part being artificially infested with the pathogen in the beginning (1st winter wheat). In the 2nd, 3rd, and 4th year, only spring crops were grown, followed by winter wheat in the 5th year. The final exploration resulted in a medium to heavy degree of infection, being significantly higher in the 80% cereal crop sequence than in the 60% and 40% sequences. Concerning the catch-crops, infection in the rye variants was equal or significantly lower than in the carefully cultivated variants without catch-crop. On the other hand, infection of the rape variants had become significantly higher than that of the catch-crop-free ones, about as heavily as in the weed-infested variants. From fall 1966 to summer 1969, the development of the saprophytic soil microflora and several other edaphic parameters had been investigated. Remarkable differences in intensity of disintegration have been observed under the influence of the main crops, cereals or potatoes, respectively, but no differences could be stated resulting from the matter of disintegration--residues of rye or rape--, which might interpret the contrasting effects on the pathogen. Too, no indication for antagonistic activity of bacteria or actinomycetes has been noted. The results are compared with known facts of Cercosporella and of other soil borne pathogens. The difference in biology of Cercosporella on the one hand, and of root infecting fungi on the other hand seems to be most important to explain the results obtained.

Actinomycetales↗

The export of amino acid in the phloem is altered in wheat plants lacking the short arm of chromosome 7B.

Grain protein content is one of the major determinants of the baking and nutritional quality of wheat. It has previously been reported that the ditelosomic line of wheat (Triticum aestivum L.) CSDT7BL, where the short arm of chromosome 7B is missing, shows a lower grain protein concentration than the normal line, but a similar grain yield. In the present paper the growth and nitrogen (N) metabolism of wheat plants cv. Chinese Spring (CS) and its ditelosomic line CSDT7BL were compared. When plants were grown to maturity in pots with different N supplements, the wild-type line showed a higher grain protein concentration and a lower straw N concentration than the ditelosomic line at every N level analysed, suggesting a deficiency in the N remobilization capacity. When 15-d-old plants were grown in a growth cabinet in pots with sand, and supplied with nutrient solutions of different nitrate concentrations, the ditelosomic line showed no differences in N uptake per unit of root dry weight, nitrate reductase activity, nitrate, total N concentration or free amino acid concentration. However, the ditelosomic line showed a decreased capacity to export amino acids in the phloem under high N, independently of the N source. This deficiency was also observed under dark-induced senescence. The diminished export of amino acids to the phloem was principally caused by a decrease in the export of Glu, Asp, and Gln. It is suggested that the decrease in grain protein concentration in the ditelosomic line is a consequence of defective export in the phloem of these amino acids.

Amino Acids↗