Lens orientalis contributes quantitative trait loci and candidate genes associated with Ascochyta blight resistance in lentil
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ABSTRACT:
PROVIDER: PRJNA736090 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR14762100_1.fastq.gz | Fastqsanger.gz | |||
| SRR14762100_2.fastq.gz | Fastqsanger.gz | |||
| SRR14762200_1.fastq.gz | Fastqsanger.gz | |||
| SRR14762200_2.fastq.gz | Fastqsanger.gz | |||
| SRR14762101_1.fastq.gz | Fastqsanger.gz |
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Project description:Using a cDNA microarray themed on Ascochyta-Pulse interaction resistance response was studied in two lentil varieties, specifically in response to A. lentis inoculation in a highly resistant (ILL7537) and highly susceptible (ILL6002) lentil variety. The experiments were conducted in reference design, where samples from mock-inoculated controls acted as references against post-inoculation samples and the plants were grown using a uniform and standardized experimental system that minimized environmental effects. Robust and high quality data was obtained through the use of three biological replicates (including a dye-swap), the inclusion of negative controls, and stringent selection criteria for differentially expressed genes including a fold change cutoff determined by self-self hybridizations, Students t-test and FDR (Fasle Discovary Rate) multiple testing correction (P<0.05). Microarray observations were validated by quantitative real time RT-PCR using the RNA from one of the bioassay used in the original microarray experiment. Ninety genes were differentially expressed in ILL7537 and 95 genes were differentially expressed in ILL6002. The expression profiles of the two varieties showed substantial difference in type and time of genes that were expressed in response to A. lentis. The resistant variety showed early up-regulation of PR proteins and other defence related genes. The susceptible genotype showed mainly down-regulation of defence related genes. The microarray experiment, the first in lentil, conducted with a small number of genes themed on Asochyta and Pulse interactions was able to identify different components of the defence mechanism by comparing the transcriptional profiles of the susceptible and resistant genotypes. This study will thus form the basis of future experiments to elaborate and corroborate the genomics of lentils defence to A. lentis. Keywords: time course, disease state analysis
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Project description:Australian lentil production is affected by several major biotic constraints including Ascochyta blight (AB), caused by Ascochyta lentis, a devastating fungal disease. Cultivation of AB resistant cultivars, alongside agronomic management including fungicide application, is the current most economically viable control strategy. However, the breakdown of AB resistance in cultivars, such as Northfield and Nipper, suggests the need for introgression of new and diverse resistance genes. Successful introgression entails an understanding of the genetic basis of resistance. In this context, a biparental mapping population derived from a cross between a recently identified AB resistant accession ILWL 180 (Lens orientalis) and a susceptible cultivar ILL 6002 was produced. A genetic linkage map was constructed from single-nucleotide polymorphism markers generated using a genotyping-by-sequencing transcript approach. Genetic dissection of the mapping population revealed a major quantitative trait loci (QTL) region nested with three QTLs on linkage group 5 and explained 9.5-11.5 percent (%) of phenotypic variance for AB resistance. Another QTL was identified on LG2 with phenotypic variance of 9.6%. The identified QTL regions harbored putative candidate genes potentially associated with defense responses to A. lentis infection. The QTL analysis and the candidate gene information are expected to contribute to the development of diagnostic markers and enable marker-assisted resistance selection in lentil breeding programmes.
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Project description:Using microarray technology and a set of chickpea (Cicer arietinum L.) unigenes, grasspea (Lathyrus sativus L.) ESTs and lentil (Lens culinaris Med.) resistance gene analogs, the ascochyta blight (Ascochyta rabiei (Pass.) L.) resistance response was studied in four chickpea genotypes, including resistant, moderately resistant, susceptible and wild relative (Cicer echinospermum L.) genotypes. The experimental system minimized environmental effects and was conducted in reference design, where samples from mock-inoculated controls acted as references against post-inoculation samples. Robust data quality was achieved through the use of three biological replicates (including a dye-swap), the inclusion of negative controls, and strict selection criteria for differentially expressed genes including a fold change cutoff determined by self-self hybridizations, Students t test and multiple testing correction (P<0.05). Microarray observations were also validated by quantitative RT-PCR. The time-course expression patterns of 756 microarray features resulted in differential expression of 97 genes in at least one genotype at one time-point. K-means clustering grouped the genes into clusters of similar observations for each genotype, and comparisons between A. rabiei-resistant and susceptible genotypes revealed potential gene 'signatures' predictive of effective A. rabiei resistance. These genes included several pathogenesis-related proteins, SNAKIN2 antimicrobial peptide, proline-rich protein, disease resistance response protein DRRG49-C, environmental stress-inducible protein, leucine-zipper protein, polymorphic antigen membrane protein, as well as several unknown proteins. The potential involvement of these genes and their pathways of induction are discussed. This study represents the first large-scale gene expression profiling in chickpea, and future work will focus on functional validation of the genes of interest. Keywords: time course disease state analysis
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