32 results on '"Douchkov, D"'
Search Results
2. Nicotianamine synthase: Gene isolation, gene transfer and application for the manipulation of plant iron assimilation
- Author
-
Douchkov, D., Herbik, A., Koch, G., Mock, H.-P., Melzer, M., Stephan, U. W., and Bäumlein, H.
- Published
- 2002
3. Increased iron efficiency in transgenic plants due to ectopic expression of nicotianamine synthase
- Author
-
Douchkov, D., Hell, R., Stephan, U. W., Bäumlein, H., Horst, W. J., editor, Schenk, M. K., editor, Bürkert, A., editor, Claassen, N., editor, Flessa, H., editor, Frommer, W. B., editor, Goldbach, H., editor, Olfs, H. -W., editor, Römheld, V., editor, Sattelmacher, B., editor, Schmidhalter, U., editor, Schubert, S., editor, v. Wirén, N., editor, and Wittenmayer, L., editor
- Published
- 2001
- Full Text
- View/download PDF
4. High throughput RNAi to study the interaction between barley and powdery mildew fungi
- Author
-
Douchkov, D., Dong, W., Zierold, U., Nowara, D., Seiffert, U., and Schweizer, P.
- Published
- 2006
- Full Text
- View/download PDF
5. The barley (Hordeum vulgare) cellulose synthase-like D2 gene (HvCslD2) mediates penetration resistance to host-adapted and nonhost isolates of the powdery mildew fungus
- Author
-
Douchkov, D., Lueck, S., Hensel, G., Kumlehn, J., Rajaraman, J., Johrde, A., Doblin, M. S., Beahan, C. T., Kopischke, M., Fuchs, R., Lipka, V., Niks, R. E., Bulone, Vincent, Chowdhury, J., Little, A., Burton, R. A., Bacic, A., Fincher, G. B., Schweizer, P., Douchkov, D., Lueck, S., Hensel, G., Kumlehn, J., Rajaraman, J., Johrde, A., Doblin, M. S., Beahan, C. T., Kopischke, M., Fuchs, R., Lipka, V., Niks, R. E., Bulone, Vincent, Chowdhury, J., Little, A., Burton, R. A., Bacic, A., Fincher, G. B., and Schweizer, P.
- Abstract
Cell walls and cellular turgor pressure shape and suspend the bodies of all vascular plants. In response to attack by fungal and oomycete pathogens, which usually breach their host's cell walls by mechanical force or by secreting lytic enzymes, plants often form local cell wall appositions (papillae) as an important first line of defence. The involvement of cell wall biosynthetic enzymes in the formation of these papillae is still poorly understood, especially in cereal crops. To investigate the role in plant defence of a candidate gene from barley (Hordeum vulgare) encoding cellulose synthase-like D2 (HvCslD2), we generated transgenic barley plants in which HvCslD2 was silenced through RNA interference (RNAi). The transgenic plants showed no growth defects but their papillae were more successfully penetrated by host-adapted, virulent as well as avirulent nonhost isolates of the powdery mildew fungus Blumeria graminis. Papilla penetration was associated with lower contents of cellulose in epidermal cell walls and increased digestion by fungal cell wall degrading enzymes. The results suggest that HvCslD2-mediated cell wall changes in the epidermal layer represent an important defence reaction both for nonhost and for quantitative host resistance against nonadapted wheat and host-adapted barley powdery mildew pathogens, respectively., QC 20161111
- Published
- 2016
- Full Text
- View/download PDF
6. The barley (Hordeum vulgare) cellulose synthase-like D2 gene (HvCslD2) mediates penetration resistance to host-adapted and nonhost isolates of the powdery mildew fungus
- Author
-
Douchkov, D, Lueck, S, Hensel, G, Kumlehn, J, Rajaraman, J, Johrde, A, Doblin, MS, Beahan, CT, Kopischke, M, Fuchs, R, Lipka, V, Niks, RE, Bulone, V, Chowdhury, J, Little, A, Burton, RA, Bacic, A, Fincher, GB, Schweizer, P, Douchkov, D, Lueck, S, Hensel, G, Kumlehn, J, Rajaraman, J, Johrde, A, Doblin, MS, Beahan, CT, Kopischke, M, Fuchs, R, Lipka, V, Niks, RE, Bulone, V, Chowdhury, J, Little, A, Burton, RA, Bacic, A, Fincher, GB, and Schweizer, P
- Abstract
Cell walls and cellular turgor pressure shape and suspend the bodies of all vascular plants. In response to attack by fungal and oomycete pathogens, which usually breach their host's cell walls by mechanical force or by secreting lytic enzymes, plants often form local cell wall appositions (papillae) as an important first line of defence. The involvement of cell wall biosynthetic enzymes in the formation of these papillae is still poorly understood, especially in cereal crops. To investigate the role in plant defence of a candidate gene from barley (Hordeum vulgare) encoding cellulose synthase-like D2 (HvCslD2), we generated transgenic barley plants in which HvCslD2 was silenced through RNA interference (RNAi). The transgenic plants showed no growth defects but their papillae were more successfully penetrated by host-adapted, virulent as well as avirulent nonhost isolates of the powdery mildew fungus Blumeria graminis. Papilla penetration was associated with lower contents of cellulose in epidermal cell walls and increased digestion by fungal cell wall degrading enzymes. The results suggest that HvCslD2-mediated cell wall changes in the epidermal layer represent an important defence reaction both for nonhost and for quantitative host resistance against nonadapted wheat and host-adapted barley powdery mildew pathogens, respectively.
- Published
- 2016
7. HyphArea - automated analysis of spatiotemporal fungal patterns
- Author
-
Baum, T., Navarro-Quezada, A., Knogge, W., Douchkov, D., Schweizer, P., Seiffert, U., and Publica
- Abstract
In phytopathology quantitative measurements are rarely used to assess crop plant disease symptoms. Instead, a qualitative valuation by eye is often the method of choice. In order to close the gap between subjective human inspection and objective quantitative results, the development of an automated analysis system that is capable of recognizing and characterizing the growth patterns of fungal hyphae in micrograph images was developed. This system should enable the efficient screening of different host-pathogen combinations (e.g., barley-. Blumeria graminis, barley-. Rhynchosporium secalis) using different microscopy technologies (e.g., bright field, fluorescence). An image segmentation algorithm was developed for gray-scale image data that achieved good results with several microscope imaging protocols. Furthermore, adaptability towards different host-pathogen systems was obtained by using a classification that is based on a genetic algorithm. The developed software sys tem was named . HyphArea, since the quantification of the area covered by a hyphal colony is the basic task and prerequisite for all further morphological and statistical analyses in this context. By means of a typical use case the utilization and basic properties of . HyphArea could be demonstrated. It was possible to detect statistically significant differences between the growth of an . R. secalis wild-type strain and a virulence mutant.
- Published
- 2011
8. A High-Throughput Gene-Silencing System for the Functional Assessment of Defense-Related Genes in Barley Epidermal Cells
- Author
-
Douchkov, D., primary, Nowara, D., additional, Zierold, U., additional, and Schweizer, P., additional
- Published
- 2005
- Full Text
- View/download PDF
9. Ectopic expression of nicotianamine synthase genes results in improved iron accumulation and increased nickel tolerance in transgenic tobacco
- Author
-
DOUCHKOV, D., primary, GRYCZKA, C., additional, STEPHAN, U. W., additional, HELL, R., additional, and BÄUMLEIN, H., additional
- Published
- 2004
- Full Text
- View/download PDF
10. Ectopic expression of nicotianamine synthase genes results in improved iron accumulation and increased nickel tolerance in transgenic tobacco.
- Author
-
Douchkov, D., Gryczka, C., Stephan, U.W., Hell, R., and Bäumlein, H.
- Subjects
- *
HEAVY metals , *HOMEOSTASIS , *AMINO acids , *PLANT nutrition , *ARABIDOPSIS thaliana , *TOBACCO , *TRANSGENIC plants - Abstract
Heavy metals are essential for basic cellular processes but toxic in higher concentrations. This requires the precise control of their intracellular concentrations, a process known as homeostasis. The metal-chelating, non-proteinogenous amino acid nicotianamine (NA) is a key component of plant metal assimilation and homeostasis. Its precise function is still unknown. Therefore, this article aims to contribute new information on thein vivofunction of NA and to evaluate its potential use for plant nutrition and crop fortification. For this purpose, a nicotianamine synthase gene ofArabidopsis thalianawas ectopically expressed in transgenic tobacco plants.The presence of extra copies of the nicotianamine synthase gene co-segregated with up to 10-fold elevated levels of NA in comparison with wild type. The increased NA level led to: (a) a significantly increased iron level in leaves of adult plants; (b) the accumulation of zinc and manganese, but not copper; (c) an improvement of the iron use efficiency in adult plants grown under iron limitation; and (d) an enhanced tolerance against up to 1 mmnickel.Taken together, the data predict that NA may be a useful tool for improved plant nutrition on adverse soils and possibly for enhanced nutritional value of leaf and seed crops. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
11. Proceedings zum Workshop “Genexpression – aktuelle Trends, neue Herausforderungen” 28.–29. Juli 2005, Universität Hannover, Hannover-Herrenhausen (Germany).
- Author
-
Douchkov, D., Dong, W., Zierold, U., Nowara, D., Seiffert, U., and Schweizer, P.
- Published
- 2006
- Full Text
- View/download PDF
12. Two pathogen loci determine Blumeria graminis f. sp. tritici virulence to wheat resistance gene Pm1a.
- Author
-
Kloppe T, Whetten RB, Kim SB, Powell OR, Lück S, Douchkov D, Whetten RW, Hulse-Kemp AM, Balint-Kurti P, and Cowger C
- Subjects
- Virulence genetics, Plant Diseases microbiology, Disease Resistance genetics, Triticum microbiology, Genome-Wide Association Study
- Abstract
Blumeria graminis f. sp. tritici (Bgt) is a globally important fungal pathogen of wheat that can rapidly evolve to defeat wheat powdery mildew (Pm) resistance genes. Despite periodic regional deployment of the Pm1a resistance gene in US wheat production, Bgt strains that overcome Pm1a have been notably nonpersistent in the United States, while on other continents, they are more widely established. A genome-wide association study (GWAS) was conducted to map sequence variants associated with Pm1a virulence in 216 Bgt isolates from six countries, including the United States. A virulence variant apparently unique to Bgt isolates from the United States was detected in the previously mapped gene AvrPm1a (BgtE-5612) on Bgt chromosome 6; an in vitro growth assay suggested no fitness reduction associated with this variant. A gene on Bgt chromosome 8, Bgt-51526, was shown to function as a second determinant of Pm1a virulence, and despite < 30% amino acid identity, BGT-51526 and BGTE-5612 were predicted to share > 85% of their secondary structure. A co-expression study in Nicotiana benthamiana showed that BGTE-5612 and BGT-51526 each produce a PM1A-dependent hypersensitive response. More than one member of a B. graminis effector family can be recognized by a single wheat immune receptor, and a two-gene model is necessary to explain virulence to Pm1a., (© 2023 The Authors. New Phytologist © 2023 New Phytologist Foundation This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.)
- Published
- 2023
- Full Text
- View/download PDF
13. High-throughput imaging of powdery mildew resistance of the winter wheat collection hosted at the German Federal ex situ Genebank for Agricultural and Horticultural Crops.
- Author
-
Hinterberger V, Douchkov D, Lueck S, Reif JC, and Schulthess AW
- Subjects
- Agriculture, Crops, Agricultural, Genotype, Triticum, Plant Breeding
- Abstract
Background: Genebanks worldwide are transforming into biodigital resource centers, providing access not only to the plant material itself but also to its phenotypic and genotypic information. Adding information for relevant traits will help boost plant genetic resources' usage in breeding and research. Resistance traits are vital for adapting our agricultural systems to future challenges., Findings: Here we provide phenotypic data for the resistance against Blumeria graminis f. sp. tritici, the causal agent of wheat powdery mildew-a substantial risk to our agricultural production. Using a modern high-throughput phenotyping system, we infected and photographed a total of 113,638 wheat leaves of 7,320 winter wheat (Triticum aestivum L.) plant genetic resources of the German Federal ex situ Genebank for Agricultural and Horticultural Crops and 154 commercial genotypes. We quantified the resistance reaction captured by images and provide them here, along with the raw images., Conclusion: This massive amount of phenotypic data, combined with already published genotypic data, also provides a valuable and unique training dataset for the development of novel genotype-based predictions as well as mapping methods., (© The Author(s) 2023. Published by Oxford University Press GigaScience.)
- Published
- 2022
- Full Text
- View/download PDF
14. Mining for New Sources of Resistance to Powdery Mildew in Genetic Resources of Winter Wheat.
- Author
-
Hinterberger V, Douchkov D, Lück S, Kale S, Mascher M, Stein N, Reif JC, and Schulthess AW
- Abstract
Genetic pathogen control is an economical and sustainable alternative to the use of chemicals. In order to breed resistant varieties, information about potentially unused genetic resistance mechanisms is of high value. We phenotyped 8,316 genotypes of the winter wheat collection of the German Federal ex situ gene bank for Agricultural and Horticultural Crops, Germany , for resistance to powdery mildew (PM), Blumeria graminis f. sp. tritici , one of the most important biotrophic pathogens in wheat. To achieve this, we used a semi-automatic phenotyping facility to perform high-throughput detached leaf assays. This data set, combined with genotyping-by-sequencing (GBS) marker data, was used to perform a genome-wide association study (GWAS). Alleles of significantly associated markers were compared with SNP profiles of 171 widely grown wheat varieties in Germany to identify currently unexploited resistance conferring genes. We also used the Chinese Spring reference genome annotation and various domain prediction algorithms to perform a domain enrichment analysis and produced a list of candidate genes for further investigation. We identified 51 significantly associated regions. In most of these, the susceptible allele was fixed in the tested commonly grown wheat varieties. Eleven of these were located on chromosomes for which no resistance conferring genes have been previously reported. In addition to enrichment of leucine-rich repeats (LRR), we saw enrichment of several domain types so far not reported as relevant to PM resistance, thus, indicating potentially novel candidate genes for the disease resistance research and prebreeding in wheat., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hinterberger, Douchkov, Lück, Kale, Mascher, Stein, Reif and Schulthess.)
- Published
- 2022
- Full Text
- View/download PDF
15. Tillage shapes the soil and rhizosphere microbiome of barley-but not its susceptibility towards Blumeria graminis f. sp. hordei.
- Author
-
Bziuk N, Maccario L, Douchkov D, Lueck S, Babin D, Sørensen SJ, Schikora A, and Smalla K
- Subjects
- Ascomycota, Plant Diseases, RNA, Ribosomal, 16S genetics, Rhizosphere, Soil, Hordeum, Microbiota
- Abstract
Long-term agricultural practices are assumed to shape the rhizosphere microbiome of crops with implications for plant health. In a long-term field experiment, we investigated the effect of different tillage and fertilization practices on soil and barley rhizosphere microbial communities by means of amplicon sequencing of 16S rRNA gene fragments from total community DNA. Differences in the microbial community composition depending on the tillage practice, but not the fertilization intensity were revealed. To examine whether these soil and rhizosphere microbiome differences influence the plant defense response, barley (cultivar Golden Promise) was grown in field or standard potting soil under greenhouse conditions and challenged with Blumeria graminis f. sp. hordei (Bgh). Amplicon sequence analysis showed that preceding tillage practice, but also aboveground Bgh challenge significantly influenced the microbial community composition. Expression of plant defense-related genes PR1b and PR17b was higher in challenged compared to unchallenged plants. The Bgh infection rates were strikingly lower for barley grown in field soil compared to potting soil. Although previous agricultural management shaped the rhizosphere microbiome, no differences in plant health were observed. We propose therefore that the management-independent higher microbial diversity of field soils compared to potting soils contributed to the low infection rates of barley., (© The Author(s) 2021. Published by Oxford University Press on behalf of FEMS.)
- Published
- 2021
- Full Text
- View/download PDF
16. A small secreted protein from Zymoseptoria tritici interacts with a wheat E3 ubiquitin ligase to promote disease.
- Author
-
Karki SJ, Reilly A, Zhou B, Mascarello M, Burke J, Doohan F, Douchkov D, Schweizer P, and Feechan A
- Subjects
- Plant Diseases, Ubiquitin-Protein Ligases genetics, Ascomycota, Triticum genetics
- Abstract
Septoria tritici blotch (STB), caused by the ascomycete fungus Zymoseptoria tritici, is a major threat to wheat production worldwide. The Z. tritici genome encodes many small secreted proteins (ZtSSPs) that are likely to play a key role in the successful colonization of host tissues. However, few of these ZtSSPs have been functionally characterized for their role during infection. In this study, we identified and characterized a small, conserved cysteine-rich secreted effector from Z. tritici which has homologues in other plant pathogens in the Dothideomycetes. ZtSSP2 was expressed throughout Z. tritici infection in wheat, with the highest levels observed early during infection. A yeast two-hybrid assay revealed an interaction between ZtSSP2 and wheat E3 ubiquitin ligase (TaE3UBQ) in yeast, and this was further confirmed in planta using bimolecular fluorescence complementation and co-immunoprecipitation. Down-regulation of this wheat E3 ligase using virus-induced gene silencing increased the susceptibility of wheat to STB. Together, these results suggest that TaE3UBQ is likely to play a role in plant immunity to defend against Z. tritici., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Experimental Biology.)
- Published
- 2021
- Full Text
- View/download PDF
17. "Macrobot": An Automated Segmentation-Based System for Powdery Mildew Disease Quantification.
- Author
-
Lück S, Strickert M, Lorbeer M, Melchert F, Backhaus A, Kilias D, Seiffert U, and Douchkov D
- Abstract
Managing plant diseases is increasingly difficult due to reasons such as intensifying the field production, climatic change-driven expansion of pests, redraw and loss of effectiveness of pesticides, rapid breakdown of the disease resistance in the field, and other factors. The substantial progress in genomics of both plants and pathogens, achieved in the last decades, has the potential to counteract this negative trend, however, only when the genomic data is supported by relevant phenotypic data that allows linking the genomic information to specific traits. We have developed a set of methods and equipment and combined them into a "Macrophenomics facility." The pipeline has been optimized for the quantification of powdery mildew infection symptoms on wheat and barley, but it can be adapted to other diseases and host plants. The Macrophenomics pipeline scores the visible powdery mildew disease symptoms, typically 5-7 days after inoculation (dai), in a highly automated manner. The system can precisely and reproducibly quantify the percentage of the infected leaf area with a theoretical throughput of up to 10000 individual samples per day, making it appropriate for phenotyping of large germplasm collections and crossing populations., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this article., (Copyright © 2020 Stefanie Lück et al.)
- Published
- 2020
- Full Text
- View/download PDF
18. Identification of novel genetic factors underlying the host-pathogen interaction between barley (Hordeum vulgare L.) and powdery mildew (Blumeria graminis f. sp. hordei).
- Author
-
Pogoda M, Liu F, Douchkov D, Djamei A, Reif JC, Schweizer P, and Schulthess AW
- Subjects
- Alleles, Ascomycota isolation & purification, Ascomycota pathogenicity, Cluster Analysis, Gene Expression Regulation, Plant, Genetic Predisposition to Disease, Genome-Wide Association Study, Genotype, Linkage Disequilibrium, Mucoproteins genetics, Mucoproteins metabolism, Phenotype, Plant Diseases genetics, Plant Proteins genetics, Plant Proteins metabolism, Seedlings genetics, Virulence genetics, Ascomycota genetics, Hordeum genetics, Host-Pathogen Interactions genetics, Plant Diseases microbiology
- Abstract
Powdery mildew is an important foliar disease of barley (Hordeum vulgare L.) caused by the biotrophic fungus Blumeria graminis f. sp. hordei (Bgh). The understanding of the resistance mechanism is essential for future resistance breeding. In particular, the identification of race-nonspecific resistance genes is important because of their regarded durability and broad-spectrum activity. We assessed the severity of powdery mildew infection on detached seedling leaves of 267 barley accessions using two poly-virulent isolates and performed a genome-wide association study exploiting 201 of these accessions. Two-hundred and fourteen markers, located on six barley chromosomes are associated with potential race-nonspecific Bgh resistance or susceptibility. Initial steps for the functional validation of four promising candidates were performed based on phenotype and transcription data. Specific candidate alleles were analyzed via transient gene silencing as well as transient overexpression. Microarray data of the four selected candidates indicate differential regulation of the transcription in response to Bgh infection. Based on our results, all four candidate genes seem to be involved in the responses to powdery mildew attack. In particular, the transient overexpression of specific alleles of two candidate genes, a potential arabinogalactan protein and the barley homolog of Arabidopsis thaliana's Light-Response Bric-a-Brac/-Tramtrack/-Broad Complex/-POxvirus and Zinc finger (AtLRB1) or AtLRB2, were top candidates of novel powdery mildew susceptibility genes., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2020
- Full Text
- View/download PDF
19. siRNA-Finder (si-Fi) Software for RNAi-Target Design and Off-Target Prediction.
- Author
-
Lück S, Kreszies T, Strickert M, Schweizer P, Kuhlmann M, and Douchkov D
- Abstract
RNA interference (RNAi) is a technique used for transgene-mediated gene silencing based on the mechanism of posttranscriptional gene silencing (PTGS). PTGS is an ubiquitous basic biological phenomenon involved in the regulation of transcript abundance and plants' immune response to viruses. PTGS also mediates genomic stability by silencing of retroelements. RNAi has become an important research tool for studying gene function by strong and selective suppression of target genes. Here, we present si-Fi , a software tool for design optimization of RNAi constructs necessary for specific target gene knock-down. It offers efficiency prediction of RNAi sequences and off-target search, required for the practical application of RNAi. si-Fi is an open-source (CC BY-SA license) desktop software that works in Microsoft Windows environment and can use custom sequence databases in standard FASTA format.
- Published
- 2019
- Full Text
- View/download PDF
20. Evolutionarily conserved partial gene duplication in the Triticeae tribe of grasses confers pathogen resistance.
- Author
-
Rajaraman J, Douchkov D, Lück S, Hensel G, Nowara D, Pogoda M, Rutten T, Meitzel T, Brassac J, Höfle C, Hückelhoven R, Klinkenberg J, Trujillo M, Bauer E, Schmutzer T, Himmelbach A, Mascher M, Lazzari B, Stein N, Kumlehn J, and Schweizer P
- Subjects
- Alleles, Base Sequence, Gene Expression Regulation, Plant, Gene Silencing, Genes, Plant, Genetic Markers, Hordeum genetics, Plant Diseases genetics, Plant Diseases microbiology, Plant Proteins metabolism, Plants, Genetically Modified, Protein Binding, Quantitative Trait, Heritable, Conserved Sequence genetics, Disease Resistance genetics, Evolution, Molecular, Gene Duplication, Host-Pathogen Interactions genetics, Poaceae genetics
- Abstract
Background: The large and highly repetitive genomes of the cultivated species Hordeum vulgare (barley), Triticum aestivum (wheat), and Secale cereale (rye) belonging to the Triticeae tribe of grasses appear to be particularly rich in gene-like sequences including partial duplicates. Most of them have been classified as putative pseudogenes. In this study we employ transient and stable gene silencing- and over-expression systems in barley to study the function of HvARM1 (for H. vulgare Armadillo 1), a partial gene duplicate of the U-box/armadillo-repeat E3 ligase HvPUB15 (for H. vulgare Plant U-Box 15)., Results: The partial ARM1 gene is derived from a gene-duplication event in a common ancestor of the Triticeae and contributes to quantitative host as well as nonhost resistance to the biotrophic powdery mildew fungus Blumeria graminis. In barley, allelic variants of HvARM1 but not of HvPUB15 are significantly associated with levels of powdery mildew infection. Both HvPUB15 and HvARM1 proteins interact in yeast and plant cells with the susceptibility-related, plastid-localized barley homologs of THF1 (for Thylakoid formation 1) and of ClpS1 (for Clp-protease adaptor S1) of Arabidopsis thaliana. A genome-wide scan for partial gene duplicates reveals further events in barley resulting in stress-regulated, potentially neo-functionalized, genes., Conclusion: The results suggest neo-functionalization of the partial gene copy HvARM1 increases resistance against powdery mildew infection. It further links plastid function with susceptibility to biotrophic pathogen attack. These findings shed new light on a novel mechanism to employ partial duplication of protein-protein interaction domains to facilitate the expansion of immune signaling networks.
- Published
- 2018
- Full Text
- View/download PDF
21. Altered Expression of Genes Implicated in Xylan Biosynthesis Affects Penetration Resistance against Powdery Mildew.
- Author
-
Chowdhury J, Lück S, Rajaraman J, Douchkov D, Shirley NJ, Schwerdt JG, Schweizer P, Fincher GB, Burton RA, and Little A
- Abstract
Heteroxylan has recently been identified as an important component of papillae, which are formed during powdery mildew infection of barley leaves. Deposition of heteroxylan near the sites of attempted fungal penetration in the epidermal cell wall is believed to enhance the physical resistance to the fungal penetration peg and hence to improve pre-invasion resistance. Several glycosyltransferase (GT) families are implicated in the assembly of heteroxylan in the plant cell wall, and are likely to work together in a multi-enzyme complex. Members of key GT families reported to be involved in heteroxylan biosynthesis are up-regulated in the epidermal layer of barley leaves during powdery mildew infection. Modulation of their expression leads to altered susceptibility levels, suggesting that these genes are important for penetration resistance. The highest level of resistance was achieved when a GT43 gene was co-expressed with a GT47 candidate gene, both of which have been predicted to be involved in xylan backbone biosynthesis. Altering the expression level of several candidate heteroxylan synthesis genes can significantly alter disease susceptibility. This is predicted to occur through changes in the amount and structure of heteroxylan in barley papillae.
- Published
- 2017
- Full Text
- View/download PDF
22. An LRR/Malectin Receptor-Like Kinase Mediates Resistance to Non-adapted and Adapted Powdery Mildew Fungi in Barley and Wheat.
- Author
-
Rajaraman J, Douchkov D, Hensel G, Stefanato FL, Gordon A, Ereful N, Caldararu OF, Petrescu AJ, Kumlehn J, Boyd LA, and Schweizer P
- Abstract
Pattern recognition receptors (PRRs) belonging to the multigene family of receptor-like kinases (RLKs) are the sensing devices of plants for microbe- or pathogen-associated molecular patterns released from microbial organisms. Here we describe Rnr8 (for Required for non-host resistance 8 ) encoding HvLEMK1, a LRR-malectin domain-containing transmembrane RLK that mediates non-host resistance of barley to the non-adapted wheat powdery mildew fungus Blumeria graminis f.sp. tritici . Transgenic barley lines with silenced HvLEMK1 allow entry and colony growth of the non-adapted pathogen, although sporulation was reduced and final colony size did not reach that of the adapted barley powdery mildew fungus B. graminis f.sp. hordei . Transient expression of the barley or wheat LEMK1 genes enhanced resistance in wheat to the adapted wheat powdery mildew fungus while expression of the same genes did not protect barley from attack by the barley powdery mildew fungus. The results suggest that HvLEMK1 is a factor mediating non-host resistance in barley and quantitative host resistance in wheat to the wheat powdery mildew fungus.
- Published
- 2016
- Full Text
- View/download PDF
23. Down-regulation of the glucan synthase-like 6 gene (HvGsl6) in barley leads to decreased callose accumulation and increased cell wall penetration by Blumeria graminis f. sp. hordei.
- Author
-
Chowdhury J, Schober MS, Shirley NJ, Singh RR, Jacobs AK, Douchkov D, Schweizer P, Fincher GB, Burton RA, and Little A
- Subjects
- Arabidopsis genetics, Hordeum microbiology, Phylogeny, Plant Epidermis cytology, Plant Epidermis metabolism, Plant Proteins genetics, Plant Proteins metabolism, RNA Interference, RNA, Messenger genetics, RNA, Messenger metabolism, Transformation, Genetic, Ascomycota physiology, Cell Wall microbiology, Down-Regulation genetics, Gene Expression Regulation, Plant, Genes, Plant, Glucosyltransferases genetics, Hordeum enzymology, Hordeum genetics
- Abstract
The recent characterization of the polysaccharide composition of papillae deposited at the barley cell wall during infection by the powdery mildew pathogen, Blumeria graminis f. sp. hordei (Bgh), has provided new targets for the generation of enhanced disease resistance. The role of callose in papilla-based penetration resistance of crop species is largely unknown because the genes involved in the observed callose accumulation have not been identified unequivocally. We have employed both comparative and functional genomics approaches to identify the functional orthologue of AtGsl5 in the barley genome. HvGsl6 (the barley glucan synthase-like 6 gene), which has the highest sequence identity to AtGsl5, is the only Bgh-induced gene among the HvGsls examined in this study. Through double-stranded RNA interference (dsRNAi)-mediated silencing of HvGsl6, we have shown that the down-regulation of HvGsl6 is associated with a lower accumulation of papillary and wound callose and a higher susceptibility to penetration of the papillae by Bgh, compared with control lines. The results indicate that the HvGsl6 gene is a functional orthologue of AtGsl5 and is involved in papillary callose accumulation in barley. The increased susceptibility of HvGsl6 dsRNAi transgenic lines to infection indicates that callose positively contributes to the barley fungal penetration resistance mechanism., (© 2016 University of Adelaide. New Phytologist © 2016 New Phytologist Trust.)
- Published
- 2016
- Full Text
- View/download PDF
24. The barley (Hordeum vulgare) cellulose synthase-like D2 gene (HvCslD2) mediates penetration resistance to host-adapted and nonhost isolates of the powdery mildew fungus.
- Author
-
Douchkov D, Lueck S, Hensel G, Kumlehn J, Rajaraman J, Johrde A, Doblin MS, Beahan CT, Kopischke M, Fuchs R, Lipka V, Niks RE, Bulone V, Chowdhury J, Little A, Burton RA, Bacic A, Fincher GB, and Schweizer P
- Subjects
- Arabidopsis genetics, Cell Wall metabolism, Gene Expression Regulation, Plant, Gene Silencing, Glucosyltransferases metabolism, Hordeum enzymology, Plant Epidermis metabolism, Plant Proteins chemistry, Plant Proteins genetics, Plants, Genetically Modified, Polysaccharides metabolism, Sequence Analysis, DNA, Ascomycota physiology, Genes, Plant, Glucosyltransferases genetics, Hordeum genetics, Hordeum microbiology, Host-Pathogen Interactions genetics, Plant Diseases microbiology
- Abstract
Cell walls and cellular turgor pressure shape and suspend the bodies of all vascular plants. In response to attack by fungal and oomycete pathogens, which usually breach their host's cell walls by mechanical force or by secreting lytic enzymes, plants often form local cell wall appositions (papillae) as an important first line of defence. The involvement of cell wall biosynthetic enzymes in the formation of these papillae is still poorly understood, especially in cereal crops. To investigate the role in plant defence of a candidate gene from barley (Hordeum vulgare) encoding cellulose synthase-like D2 (HvCslD2), we generated transgenic barley plants in which HvCslD2 was silenced through RNA interference (RNAi). The transgenic plants showed no growth defects but their papillae were more successfully penetrated by host-adapted, virulent as well as avirulent nonhost isolates of the powdery mildew fungus Blumeria graminis. Papilla penetration was associated with lower contents of cellulose in epidermal cell walls and increased digestion by fungal cell wall degrading enzymes. The results suggest that HvCslD2-mediated cell wall changes in the epidermal layer represent an important defence reaction both for nonhost and for quantitative host resistance against nonadapted wheat and host-adapted barley powdery mildew pathogens, respectively., (© 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.)
- Published
- 2016
- Full Text
- View/download PDF
25. Discovery of genes affecting resistance of barley to adapted and non-adapted powdery mildew fungi.
- Author
-
Douchkov D, Lück S, Johrde A, Nowara D, Himmelbach A, Rajaraman J, Stein N, Sharma R, Kilian B, and Schweizer P
- Subjects
- Adaptation, Biological, Gene Expression Regulation, Plant, Gene Silencing, Genes, Plant, Hordeum microbiology, Plant Diseases genetics, Quantitative Trait Loci, Ascomycota physiology, Disease Resistance, Hordeum genetics, Hordeum immunology, Plant Diseases microbiology
- Abstract
Background: Non-host resistance, NHR, to non-adapted pathogens and quantitative host resistance, QR, confer durable protection to plants and are important for securing yield in a longer perspective. However, a more targeted exploitation of the trait usually possessing a complex mode of inheritance by many quantitative trait loci, QTLs, will require a better understanding of the most important genes and alleles., Results: Here we present results from a transient-induced gene silencing, TIGS, approach of candidate genes for NHR and QR in barley against the powdery mildew fungus Blumeria graminis. Genes were selected based on transcript regulation, multigene-family membership or genetic map position. Out of 1,144 tested RNAi-target genes, 96 significantly affected resistance to the non-adapted wheat- or the compatible barley powdery mildew fungus, with an overlap of four genes. TIGS results for QR were combined with transcript regulation data, allele-trait associations, QTL co-localization and copy number variation resulting in a meta-dataset of 51 strong candidate genes with convergent evidence for a role in QR., Conclusions: This study represents an initial, functional inventory of approximately 3% of the barley transcriptome for a role in NHR or QR against the powdery mildew pathogen. The discovered candidate genes support the idea that QR in this Triticeae host is primarily based on pathogen-associated molecular pattern-triggered immunity, which is compromised by effector molecules produced by the compatible pathogen. The overlap of four genes with significant TIGS effects both in the NHR and QR screens also indicates shared components for both forms of durable pathogen resistance.
- Published
- 2014
- Full Text
- View/download PDF
26. The conserved oligomeric Golgi complex is involved in penetration resistance of barley to the barley powdery mildew fungus.
- Author
-
Ostertag M, Stammler J, Douchkov D, Eichmann R, and Hückelhoven R
- Subjects
- Plant Diseases genetics, Plant Immunity genetics, Ascomycota pathogenicity, Golgi Apparatus metabolism, Hordeum metabolism, Hordeum microbiology, Plant Diseases microbiology, Plant Immunity physiology
- Abstract
Membrane trafficking is vital to plant development and adaptation to the environment. It is suggested that post-Golgi vesicles and multivesicular bodies are essential for plant defence against directly penetrating fungal parasites at the cell wall. However, the actual plant proteins involved in membrane transport for defence are largely unidentified. We applied a candidate gene approach and single cell transient-induced gene silencing for the identification of membrane trafficking proteins of barley involved in the response to the fungal pathogen Blumeria graminis f.sp. hordei. This revealed potential components of vesicle tethering complexes [putative exocyst subunit HvEXO70F-like and subunits of the conserved oligomeric Golgi (COG) complex] and Golgi membrane trafficking (COPIγ coatomer and HvYPT1-like RAB GTPase) as essential for resistance to fungal penetration into the host cell., (© 2012 THE AUTHORS. MOLECULAR PLANT PATHOLOGY © 2012 BSPP AND BLACKWELL PUBLISHING LTD.)
- Published
- 2013
- Full Text
- View/download PDF
27. HyphArea--automated analysis of spatiotemporal fungal patterns.
- Author
-
Baum T, Navarro-Quezada A, Knogge W, Douchkov D, Schweizer P, and Seiffert U
- Subjects
- Microscopy, Fluorescence, Ascomycota pathogenicity, Hordeum cytology, Hordeum microbiology, Plant Diseases microbiology, Software
- Abstract
In phytopathology quantitative measurements are rarely used to assess crop plant disease symptoms. Instead, a qualitative valuation by eye is often the method of choice. In order to close the gap between subjective human inspection and objective quantitative results, the development of an automated analysis system that is capable of recognizing and characterizing the growth patterns of fungal hyphae in micrograph images was developed. This system should enable the efficient screening of different host-pathogen combinations (e.g., barley-Blumeria graminis, barley-Rhynchosporium secalis) using different microscopy technologies (e.g., bright field, fluorescence). An image segmentation algorithm was developed for gray-scale image data that achieved good results with several microscope imaging protocols. Furthermore, adaptability towards different host-pathogen systems was obtained by using a classification that is based on a genetic algorithm. The developed software system was named HyphArea, since the quantification of the area covered by a hyphal colony is the basic task and prerequisite for all further morphological and statistical analyses in this context. By means of a typical use case the utilization and basic properties of HyphArea could be demonstrated. It was possible to detect statistically significant differences between the growth of an R. secalis wild-type strain and a virulence mutant., (Copyright © 2010 Elsevier GmbH. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
28. Convergent evidence for a role of WIR1 proteins during the interaction of barley with the powdery mildew fungus Blumeria graminis.
- Author
-
Douchkov D, Johrde A, Nowara D, Himmelbach A, Lueck S, Niks R, and Schweizer P
- Subjects
- Amino Acid Sequence, Ascomycota growth & development, Gene Expression Regulation, Plant genetics, Gene Expression Regulation, Plant physiology, Hordeum genetics, Molecular Sequence Data, Phylogeny, Plant Epidermis genetics, Plant Epidermis metabolism, Plant Epidermis microbiology, Plant Proteins chemistry, Plant Proteins classification, Plant Proteins genetics, Quantitative Trait Loci, RNA Interference, Sequence Homology, Amino Acid, Ascomycota physiology, Hordeum metabolism, Hordeum microbiology, Plant Proteins metabolism
- Abstract
Pathogen attack triggers a multifaceted defence response in plants that includes the accumulation of pathogenesis-related proteins and their corresponding transcripts. One of these transcripts encodes for WIR1, a small glycine- and proline-rich protein of unknown function that appears to be specific to grass species. Here we describe members of the HvWIR1 multigene family of barley with respect to phylogenetic relationship, transcript regulation, co-localization with quantitative trait loci for resistance to the barley powdery mildew fungus Blumeria graminis (DC.) E.O. Speer f.sp. hordei, the association of single nucleotide polymorphisms or gene haplotypes with resistance, as well as phenotypic effects of gene silencing by RNAi. HvWIR1 is encoded by a multigene family of moderate complexity that splits up into two major clades, one of those being also represented by previously described cDNA sequences from wheat. All analysed WIR1 transcripts accumulated in response to powdery mildew attack in leaves and all mapped WIR1 genes were associated with quantitative trait loci for resistance to B. graminis. Moreover, single nucleotide polymorphisms or haplotypes of WIR1 members were associated with quantitative resistance of barley to B. graminis, and transient WIR1 gene silencing affected the interaction of epidermal cells with the pathogen. The presented data provide convergent evidence for a role of the HvWIR1a gene and possibly other family members, during the interaction of barley with B. graminis., (Copyright © 2010 Elsevier GmbH. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
29. HIGS: host-induced gene silencing in the obligate biotrophic fungal pathogen Blumeria graminis.
- Author
-
Nowara D, Gay A, Lacomme C, Shaw J, Ridout C, Douchkov D, Hensel G, Kumlehn J, and Schweizer P
- Subjects
- Gene Expression Regulation, Fungal, Hordeum genetics, Molecular Sequence Data, Plants, Genetically Modified genetics, Plants, Genetically Modified microbiology, RNA, Antisense metabolism, RNA, Double-Stranded metabolism, RNA, Fungal genetics, RNA, Plant metabolism, Triticum genetics, Ascomycota genetics, Hordeum microbiology, Host-Pathogen Interactions, RNA Interference, Triticum microbiology
- Abstract
Powdery mildew fungi are obligate biotrophic pathogens that only grow on living hosts and cause damage in thousands of plant species. Despite their agronomical importance, little direct functional evidence for genes of pathogenicity and virulence is currently available because mutagenesis and transformation protocols are lacking. Here, we show that the accumulation in barley (Hordeum vulgare) and wheat (Triticum aestivum) of double-stranded or antisense RNA targeting fungal transcripts affects the development of the powdery mildew fungus Blumeria graminis. Proof of concept for host-induced gene silencing was obtained by silencing the effector gene Avra10, which resulted in reduced fungal development in the absence, but not in the presence, of the matching resistance gene Mla10. The fungus could be rescued from the silencing of Avra10 by the transient expression of a synthetic gene that was resistant to RNA interference (RNAi) due to silent point mutations. The results suggest traffic of RNA molecules from host plants into B. graminis and may lead to an RNAi-based crop protection strategy against fungal pathogens.
- Published
- 2010
- Full Text
- View/download PDF
30. A set of modular binary vectors for transformation of cereals.
- Author
-
Himmelbach A, Zierold U, Hensel G, Riechen J, Douchkov D, Schweizer P, and Kumlehn J
- Subjects
- Gene Expression, Genetic Markers, Molecular Sequence Data, Phenotype, Plants, Genetically Modified, Promoter Regions, Genetic, RNA Interference, Genetic Engineering, Genetic Vectors, Hordeum genetics, Transformation, Genetic, Triticum genetics
- Abstract
Genetic transformation of crop plants offers the possibility of testing hypotheses about the function of individual genes as well as the exploitation of transgenes for targeted trait improvement. However, in most cereals, this option has long been compromised by tedious and low-efficiency transformation protocols, as well as by the lack of versatile vector systems. After having adopted and further improved the protocols for Agrobacterium-mediated stable transformation of barley (Hordeum vulgare) and wheat (Triticum aestivum), we now present a versatile set of binary vectors for transgene overexpression, as well as for gene silencing by double-stranded RNA interference. The vector set is offered with a series of functionally validated promoters and allows for rapid integration of the desired genes or gene fragments by GATEWAY-based recombination. Additional in-built flexibility lies in the choice of plant selectable markers, cassette orientation, and simple integration of further promoters to drive specific expression of genes of interest. Functionality of the cereal vector set has been demonstrated by transient as well as stable transformation experiments for transgene overexpression, as well as for targeted gene silencing in barley.
- Published
- 2007
- Full Text
- View/download PDF
31. An integrated approach for the comparative analysis of a multigene family: the nicotianamine synthase genes of barley.
- Author
-
Perovic D, Tiffin P, Douchkov D, Bäumlein H, and Graner A
- Subjects
- Amino Acid Sequence, Chromosome Mapping, Molecular Sequence Data, Alkyl and Aryl Transferases genetics, Genomics, Hordeum enzymology, Hordeum genetics, Multigene Family
- Abstract
Recent genomic projects reveal that about half of the gene repertoire in plant genomes is made up by multigene families. In this paper, a set of structural and phylogenetic analyses have been applied to compare the differently sized nicotianamine synthase (NAS) gene families in barley and rice. Nicotianamine acts as a chelator of iron and other heavy metals and plays a key role in uptake, phloem transport and cytoplasmic distribution of iron, challenging efforts for the breeding of iron-efficient crop plants. Nine barley NAS genes have been mapped, and co-linearity of flanking genes in barley and rice was determined. The combined analyses reveal that the NAS multigene family members in barley originated through at least one duplication event that occurred before the divergence of rice and barley. Additional duplications appear to have occurred within each of the species. Although we detected no evidence for positive selection of recently duplicated genes within species, codon-based tests revealed evidence for positive selection having contributed to the divergence of some amino acids. The integrated comparative and phylogenetic analysis improved our current view of NAS gene family evolution, might facilitate the functional characterization of individual members and is applicable to other multigene families.
- Published
- 2007
- Full Text
- View/download PDF
32. Stable expression of a defense-related gene in wheat epidermis under transcriptional control of a novel promoter confers pathogen resistance.
- Author
-
Altpeter F, Varshney A, Abderhalden O, Douchkov D, Sautter C, Kumlehn J, Dudler R, and Schweizer P
- Subjects
- Ascomycota growth & development, Electrophoresis, Polyacrylamide Gel, Gene Expression Regulation, Plant, Immunity, Innate genetics, Oxidoreductases genetics, Oxidoreductases metabolism, Peroxidase genetics, Peroxidase metabolism, Plant Diseases microbiology, Plant Epidermis microbiology, Plant Leaves genetics, Plant Leaves metabolism, Plant Leaves microbiology, Plants, Genetically Modified, RNA, Messenger genetics, RNA, Messenger metabolism, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Reverse Transcriptase Polymerase Chain Reaction, Transcription, Genetic genetics, Triticum metabolism, Triticum microbiology, Plant Diseases genetics, Plant Epidermis genetics, Promoter Regions, Genetic genetics, Triticum genetics
- Abstract
Tissue-specific or regulated expression of transgenes is desirable in order to prevent pleiotropic side effects of putatively harmful transgene products as well as loss of energy resources due to unnecessary accumulation of transgene products. Epidermis-specific expression would be useful for many defense-related genes directed against attack by fungal pathogens that enter the plant body by direct penetration through the epidermis. In an approach to enhance resistance of wheat to the powdery mildew fungus Blumeria graminis f.sp. tritici, a novel epidermis-specific promoter was developed and used for expression of two defense-related genes. A 2.3 kb fragment of the wheat GstA1 promoter in combination with an intron-containing part of the wheat WIR1a gene was found to drive strong and constitutive transient expression in wheat epidermis. This promoter-intron combination was used for overexpression of oxalate oxidase 9f-2.8 and TaPERO peroxidase, two defense-related wheat genes expressed in inner leaf tissues. Expression studies of several transgenic lines by in situ oxalate-oxidase staining, RNA and protein blot analyses, as well as real-time PCR, demonstrated strong and constitutive transgene expression in the shoot epidermis. Transient as well as stable over-expression of the TaPERO peroxidase gene in wheat epidermis under the control of the GstA1i promoter resulted in enhanced resistance against Blumeria graminis f.sp. tritici, whereas oxalate-oxidase overexpression had no effect in either system. The data suggest that the wheat GstA1 promoter in combination with the WIR1a intron is useful for transgenic approaches to fungal disease resistance in cereals.
- Published
- 2005
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.