Dynamics of tolerant potato-potato virus Y interaction
ABSTRACT: Time series response of potato cv. Désirée, which is tolerant to PVY infection, was analysed in both inoculated as well as upper non-inoculated leaves. Additionally, transgenic plants deficient in accumulation of salicylic acid (NahG- Désirée) were studied in the same setting. 2 genotypes, mock and virus inoculated plants, inoculated (1-7dpi) and non-inoculated (1-11 dpi) leaves; one-colour design
Project description:Our aim was to investigate involvement of salicylic acid signalling in Ny-mediated hypersensistive response by comparison of transcriptomic response to the Potato Virus Y in potato plants of genotypes Rywal, bearing Ny-1 allele and NahG-Rywal, unable to accumulate salicylic acid in three time points (1, 3 and 6 days post inoculation) after viral inoculation. Inoculated leaves of mock- and virus- inoculated plants (four each) were sampled 1, 3 and 6 days after inoculation (dpi). POCI microarrays were used (one-color design)
Project description:Diploid potato plants expressing non-necrotic resistance (nnr) or which were susceptible (S) to Potato virus A (PVA) were inoculated with PVA and samples from the inoculated leaves were collected 0h, 6h, 9h, 12h, 24h and 48h post-inoculation. Control plants were mock-inoculated. The comparisons on the microarrays were made nnr vs. S and nnr vs. nnr-mock. Abbreviations: R = resistant (same as ra and nnr), RM = resistant-mock, S = susceptible, A, B and C are three independent experiments.
Project description:The late blight pathogen, Phytophthora infestans has a broad host range within the Solanaceae family, including yellow potato (Solanum phureja). The disease caused by P. infestans in S. phureja is poorly understood and is a major concern in Colombia. Expressed Sequence Tag (EST) libraries obtained from a normalized library constructed from healthy plant tissue revealed high levels of sequence similarity between S. phureja and S. tuberosum. Then, utilizing Serial Analysis of Gene Expression and high-throughput sequencing (SAGE-Solexa), we characterized yellow potato gene expression during infection by P. infestans. Four-week-old yellow potato plants were inoculated with P. infestans and were collected at 12 and 72 hours post inoculation for RNA extraction. We detected differentially expressed genes by comparing inoculated to non-inoculated and resistant to susceptible plants. The discovery and characterization of the proteins mediating this host–pathogen interaction enable the understanding of the pathosystem and is the key for developing resistant plants. Keywords: SAGE-Solexa, inoculation response, transcript profiling, Solanum phureja, Phytophthora infestans Four-week-old yellow potato (Solanum phureja) plants were inoculated with Phytophthora infestans and were collected and flash frozen in liquid nitrogen at 12 and 72 hours post inoculation, as well as mock inoculated, for RNA extraction. 2 yellow potato cultivars (resistant and susceptible) were used for each experiment. Mock inoculated plants were collected in each replicate. RNA obtained from each of the three biological replicates was pooled to obtain a single RNA sample for each timepoint X cultivar combination. A total of 6 different SAGE libraries were thus obtained. For all libraries, Illumina sequencing was performed at Canada´s Michael Smith Genome Sciences Centre.
Project description:Potato Late blight is one the most important crop diseases worldwide. Even though potato has been studied for many years, the potato disease late blight still has a huge negative effect on the potato production. A total of three commercially available field potato cultivars of different resistance to late blight infection: Kuras (moderate), Sarpo Mira (highly resistant) and Bintje (very suseptable) under controlled green house growing conditions innoculated with a diversity of P. infestans populations. We used label-free quantitative proteomics to investigate the infection with P. infestans in a time-course study over 258 hours. Several key issues limits proteome analysis of potato leaf tissue4–6. Firstly, the immense complexity of the plant proteome which is further complicated by the presence of highly abundant proteins, such as ribulose bisphosphate carboxylase/oxygenase (RuBisCO). Secondly, plant leaf and potato in particular contain abundant levels amounts of phenols and polyphenols which hinder or, unless precautions are taken, completely prevent a successful protein extraction.
Project description:In the present study molecular interactions between potato plants, Colorado potato beetle (CPB) larvae and Potato virus YNTN (PVYNTN) were investigated by analyzing gene expression in potato leaves. mRNA samples of secondary PVYNTN-infected (CPB_PVY) and healthy potato plants (CPB_H) cultivar Igor and of RNAi coi1-silenced (CPB_coi1) and non-transformed (CPB_NT) potato plants cultivar Desiree collected 24 h post CPB infestation and respective control non-infested samples (CONT_PVY, CONT_H, CONT_coi1, CONT_NT).
Project description:In order to analyze the production of small RNA (sRNA) by viroids upon infecting the plants, the tomato plants (Lycopersicum esculentum cv. Rutgers) were inoculated with the variants of Potato spindle tuber viroid (PSTVd). After 21-days of post inoculation, total RNA was extracted and subjected for deep-sequencing using Illumina platform. The primers were trimmed and only 21- to 24-nt long small RNAs were filtered after quality check of the raw data. The filtered 21- to 24-nt was mapped against the genomic and antigenomic strands of the respective PSTVd variants using standard pattern matching algorithm. The profiling of viroid derived sRNA (vd-sRNA) revealed that the viroids are susceptible to host RNA silencing mechanism. Evaluation of the vd-sRNA production in PSTVd infected tomato plants by high-throughput sequencing of small RNAs.
Project description:A comparison between two suppression subtractive hybridization cDNA libraries. One library is enriched with cDNAs from Potato virus A (PVA) resistant potato genotypes (non-necrotic resistance, nnr) 24h post-inoculation and the other library from PVA-susceptible potato 24h post-inoculation.
Project description:Feather pecking is a major welfare problem in egg production. It may be caused by genetic, physiological and environmental factors. The main aim of this study was to uncover variability in gene expression between individuals from high (HFP) and for low feather pecking (LFP) line using Chicken Gene Expression Microarrays (Agilent Technologies). Samples were assorted to two groups, each containing 9 biological replicates from high feather pecking (HFP) and low feather pecking (LFP) line.
Project description:To identify the non-host responsive genes, a time-series based global expression profiling was performed in barley using Agilent chip. Seven-day-old barley plants of cv. Vada were inoculated with spore density of 50-80 conidia mm-2 of Bgh or Bgt, and the abaxial epidermis of inoculated primary leaves or from non-inoculated control leaves was peeled 6, 12, 24 and 74 h after inoculation. Total RNA was extracted by using the RNeasy plant mini kit with on-Column DNase digestion (Qiagen, Hilden, Germany), and hybridized to a 44K Agilent oligonucleotide custom array of barley.
Project description:To identify the nonhost resistance genes, a time-series based global expression profiling was performed in wheat using Agilent gene expression 44K microarray array. Seven-day-old wheat plants of cv. Renan was inoculated with spore density of 50-80 conidia mm-2 of adapted (Bgt) or non-adapted (Bgh) powdery mildew pathogen, and the abaxial epidermis of inoculated primary leaves or from non-inoculated control leaves was peeled at 6, 12, 24 and 74 h after inoculation. Total RNA was extracted by using the RNeasy plant mini kit with on-Column DNase digestion (Qiagen, Hilden, Germany), and hybridized to a 44K Agilent oligonucleotide custom array of wheat.