- Publication type
- Meeting Abstract MeSH
Biochemical characteristics of patatin proteins purified by ion-exchange and affinity chromatography from tubers of 20 potato cultivars were studied to evaluate their genotype differences with respect to utility groups, table potato cultivars (TPCs) and processing potato cultivars (PPCs). Both groups of cultivars showed similar values of protein content in dry matter (3.98-7.39%) and of patatin relative abundance (5.40-35.40%). Three mass levels (∼40.6, 41.8, and 42.9 kDa) of purified patatins were found by MALDI-TOF MS within all cultivars. Differences among mass levels corresponding with the mass of sugar antenna (∼1.2 kDa) confirmed the previous concept of different glycosylation extentsin patatin proteins. It was showed that the individual types of patatin varying in their masses occur in the patatin family in a ratio specific for each of the cultivars, with the lowest mass type being the major one. Electrophoretic analyses demonstrated wide cultivar variability in number of patatin forms. Especially 2D-PAGE showed 17-23 detected protein spots independently on the utility group. Specific lipid acyl hydrolase (LAH) activity of purified patatins from the individual tested cultivars varied between 0.92 and 5.46 μmol/(min mg). Patatin samples within most of the TPCs exhibited higher values of specific LAH activity than samples of PPCs. It may be supposed that individual patatin forms do not have similar physiological roles.
- MeSH
- Species Specificity MeSH
- Glycosylation MeSH
- Plant Tubers chemistry MeSH
- Carboxylic Ester Hydrolases chemistry isolation & purification metabolism MeSH
- Molecular Weight MeSH
- Protein Isoforms chemistry isolation & purification metabolism MeSH
- Plant Proteins analysis chemistry isolation & purification metabolism MeSH
- Solanum tuberosum chemistry MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
Cereal crops such as wheat and maize have large repeat-rich genomes that make cloning of individual genes challenging. Moreover, gene order and gene sequences often differ substantially between cultivars of the same crop species. A major bottleneck for gene cloning in cereals is the generation of high-quality sequence information from a cultivar of interest. In order to accelerate gene cloning from any cropping line, we report 'targeted chromosome-based cloning via long-range assembly' (TACCA). TACCA combines lossless genome-complexity reduction via chromosome flow sorting with Chicago long-range linkage to assemble complex genomes. We applied TACCA to produce a high-quality (N50 of 9.76 Mb) de novo chromosome assembly of the wheat line CH Campala Lr22a in only 4 months. Using this assembly we cloned the broad-spectrum Lr22a leaf-rust resistance gene, using molecular marker information and ethyl methanesulfonate (EMS) mutants, and found that Lr22a encodes an intracellular immune receptor homologous to the Arabidopsis thaliana RPM1 protein.
- MeSH
- Biotechnology MeSH
- Genetic Linkage MeSH
- Edible Grain genetics MeSH
- Cloning, Molecular methods MeSH
- Triticum genetics MeSH
- Genes, Plant genetics MeSH
- Publication type
- Journal Article MeSH
Unraveling and exploiting mechanisms of disease resistance in cereal crops is currently limited by their large repeat-rich genomes and the lack of genetic recombination or cultivar (cv)-specific sequence information. We cloned the first leaf rust resistance gene Rph1 (Rph1a) from cultivated barley (Hordeum vulgare) using "MutChromSeq," a recently developed molecular genomics tool for the rapid cloning of genes in plants. Marker-trait association in the CI 9214/Stirling doubled haploid population mapped Rph1 to the short arm of chromosome 2H in a physical region of 1.3 megabases relative to the barley cv Morex reference assembly. A sodium azide mutant population in cv Sudan was generated and 10 mutants were confirmed by progeny-testing. Flow-sorted 2H chromosomes from Sudan (wild type) and six of the mutants were sequenced and compared to identify candidate genes for the Rph1 locus. MutChromSeq identified a single gene candidate encoding a coiled-coil nucleotide binding site Leucine-rich repeat (NLR) receptor protein that was altered in three different mutants. Further Sanger sequencing confirmed all three mutations and identified an additional two independent mutations within the same candidate gene. Phylogenetic analysis determined that Rph1 clustered separately from all previously cloned NLRs from the Triticeae and displayed highest sequence similarity (89%) with a homolog of the Arabidopsis (Arabidopsis thaliana) disease resistance protein 1 protein in Triticum urartu In this study we determined the molecular basis for Rph1-mediated resistance in cultivated barley enabling varietal improvement through diagnostic marker design, gene editing, and gene stacking technologies.
Control of common scab disease can be reached by resistant cultivars or suppressive soils. Both mechanisms are likely to translate into particular potato microbiome profiles, but the relative importance of each is not known. Here, microbiomes of bulk and tuberosphere soil and of potato periderm were studied in one resistant and one susceptible cultivar grown in a conducive and a suppressive field. Disease severity was suppressed similarly by both means yet, the copy numbers of txtB gene (coding for a pathogenicity determinant) were similar in both soils but higher in periderms of the susceptible cultivar from conducive soil. Illumina sequencing of 16S rRNA genes for bacteria (completed by 16S rRNA microarray approach) and archaea, and of 18S rRNA genes for micro-eukarytes showed that in bacteria, the more important was the effect of cultivar and diversity decreased from resistant cultivar to bulk soil to susceptible cultivar. The major changes occurred in proportions of Actinobacteria, Chloroflexi, and Proteobacteria. In archaea and micro-eukaryotes, differences were primarily due to the suppressive and conducive soil. The effect of soil suppressiveness × cultivar resistance depended on the microbial community considered, but differed also with respect to soil and plant nutrient contents particularly in N, S and Fe.
- MeSH
- Actinobacteria classification genetics growth & development pathogenicity MeSH
- Archaea classification genetics growth & development pathogenicity MeSH
- Chloroflexi classification genetics growth & development pathogenicity MeSH
- Nitrogen metabolism pharmacology MeSH
- Eukaryotic Cells metabolism MeSH
- Virulence Factors genetics metabolism MeSH
- Genotyping Techniques MeSH
- Microbiota genetics MeSH
- Disease Susceptibility immunology MeSH
- Plant Diseases immunology microbiology MeSH
- Disease Resistance drug effects MeSH
- Proteobacteria classification genetics growth & development pathogenicity MeSH
- Soil Microbiology * MeSH
- RNA, Ribosomal, 16S genetics MeSH
- RNA, Ribosomal, 18S genetics MeSH
- Sulfur metabolism pharmacology MeSH
- Solanum tuberosum drug effects immunology microbiology MeSH
- Iron metabolism pharmacology MeSH
- Crops, Agricultural MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
In view of the threat posed by climate change, we studied the influence of temperature, precipitation, cultivar characteristics, and technical management measures on the occurrence of phytopathogenic fungi in wheat during 2009-2013. This work involved experiments at two sites differing in average temperatures and precipitation. Temperature and precipitation appear to influence differences in the spectrum of phytopathogenic fungi at the individual sites. In 2009 (the warmest year), Alternaria triticina was dominant. In 2010 (having the smallest deviations from the average for individual years), Septoria tritici dominated. In 2011, Puccinia triticina was most prominent, while in 2012, the genus Drechslera (Pyrenophora) and in 2013, S. tritici and Drechslera tritici-repentis (DTR) dominated. Temperature and precipitation levels in the individual spring months (warmer March to May) played a large role, especially for the leaf rust P. triticina in 2011. A change of only 1 °C with different precipitation during a year played a significant role in changing wheat's fungal spectrum. Cluster analysis showed the differences between single pathogenic fungi on wheat in a single year due to temperature and precipitation. Alternaria abundance was strongly influenced by year (p < 0.001) while locality was significant only in certain years (2012, 2013; p = 0.004 and 0.015, respectively). The same factors were revealed to be significant in the case of Puccinia, but locality played a role (p < 0.001) in different years (2011, 2013). The abundance of S. tritici and Pyrenophora tritici-repentis (Drechslera tritici-repentis) was influenced only by year (p < 0.001).
- MeSH
- Rain MeSH
- Fungi isolation & purification MeSH
- Plant Diseases microbiology MeSH
- Triticum microbiology MeSH
- Temperature MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
- Publication type
- Meeting Abstract MeSH
In Italy a particularly valuable chestnut is "Marrone di Roccadaspide", a protected geographical indication (PGI) product, deriving from a Castanea sativa cultivar, typical of Salerno province in Campania region. As chestnut industrial processes yield a large amount of shell by-products, in this study the possibility to retrain this waste food as potential source of bioactives was investigated. The ability of "Marrone di Roccadaspide" shell MeOH extract to modulate the pro-inflammatory transcriptional factor NF-κB after LPS stimulation, along with the antioxidant activity by a cell-based in vitro test, were evaluated. To correlate the NF-κB inhibition (67.67% at 5 μg/mL) and the strong antioxidant activity to the chemical composition, an analytical approach based on LC-ESI/LTQOrbitrap/MS/MSn along with NMR characterization of isolated compounds was developed. The identification of hydrolysable and condensed tannins, along with flavonoids, phenol glucosides, ellagic acid derivatives, and triterpenoids was accomplished. The most representative compounds were quantitatively analyzed by LC-ESI/QTrap/MS/MS, showing bartogenic acid as the compound occurring in the highest amount (103.08 mg/100 g shells). With the aim to explore the possibility to employ chestnut shells as suitable source of bioactives for the preparation of functional ingredients, the chemical composition and the antioxidant activity of "eco-friendly" extracts (EtOH and EtOH:H2O 7:3) was finally evaluated, showing a high superimposability of the EtOH:H2O (7:3) extract to the MeOH extract.
- MeSH
- Antioxidants analysis MeSH
- Fagaceae chemistry MeSH
- Phenols analysis MeSH
- Flavonoids analysis MeSH
- Glucosides analysis MeSH
- Evaluation Studies as Topic MeSH
- Hydrolyzable Tannins analysis MeSH
- Calibration MeSH
- Ellagic Acid analysis MeSH
- Humans MeSH
- NF-kappa B metabolism MeSH
- Nuts chemistry MeSH
- Proanthocyanidins analysis MeSH
- Cell Proliferation MeSH
- Rationalization * MeSH
- Reactive Oxygen Species MeSH
- Plant Extracts analysis MeSH
- Tandem Mass Spectrometry MeSH
- Tannins analysis MeSH
- Triterpenes analysis MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Italy MeSH
In a three-year period (2000-2002) total tocols (tocopherols and tocotrienols), content of vitamin E and its isomers (alpha-, beta+gamma-, delta-tocopherols and tocotrienols) were assessed in grain of 13 barley genotypes. The highest content of tocols (60.3-67.6 mg kg(-1)) and content of vitamin E (Vitamin E equivalent-18.0-20.1 mg kg(-1)) were determined in the waxy varieties Wanubet, Wabet, and Washonubet. Standard varieties, i.e. of a malting type (Krona and Kompakt), had statistically significantly lower content of tocols (49.9 and 53.6 mg kg(-1)) and vitamin E (15.7-16.1 mg kg(-1)) compared to the waxy varieties. The hulless waxy variety Washonubet had statistically significantly higher total content of tocols (67.6 mg kg(-1)) and alpha- tocotrienols isomer (42.1 mg kg(-1)) versus all other genotypes in the set. Chemical treatment and fertilization statistically significantly increased the content of tocols (by 4.7 mg kg(-1)), vitamin E (by 1.9 mg kg(-1)), isomer alpha-tocopherol (by 0.9 mg kg(-1)) and isomer alpha- tocotrienols (by 3.3 mg kg(-1)). The average values of alpha-tocopherols and alpha-tocotrienols in the set were 6.7 mg kg(-1) and 29.7 mg kg(-1), respectively. Some of the reciprocal lines created by us from the malting and waxy varieties are suitable for food use for high contents of all tocopherols and alpha-tocotrienols.
- MeSH
- Financing, Organized MeSH
- Plants, Genetically Modified MeSH
- Genotype MeSH
- Hordeum genetics chemistry MeSH
- Humans MeSH
- Nutritive Value MeSH
- Tocopherols analysis MeSH
- Tocotrienols analysis MeSH
- Agriculture methods MeSH
- Check Tag
- Humans MeSH
- Publication type
- Comparative Study MeSH
