Project description:Phytophthora spp. encode large sets of effector proteins and distinct populations of small RNAs (sRNAs). Reports suggest that pathogen-derived sRNAs can modulate the expression of plant defense genes. The experiments reported here were designed to shed light on impact of sRNAs in the potato-P. infestans interaction. We used the Argonaute or Ago1 from P. infestans tagged with GFP transformed into the 88069 strain to infect potato cv. Bintje plants. Collected leaf materials were used in co-immunoprecipitation experiments together with P. infestans harboring GFP (control GFP) and P. infestans mycelia grown on media (control mycelia). These three materials were sequenced at a Ion Proton platform. The reads length of 8-38 nt were adaptor-trimmed and mapped to the P. infestans genome and the Solanom tuberosum genome v4.04. Both P. infestans-associated and potato derived sRNAs were identified.
Project description:Deep sequencing of small RNAs from three Phytophthora species, P. infestans, P. ramorum and P. sojae, was done to systematically analyze small RNA-generating components of Phytophthora genomes. We found that each species produces two distinct small RNA populations that are predominantly 21- or 25-nucleotides long. We present evidence that 25-nucleotide small RNAs are short-interfering RNAs that silence repetitive genetic elements. In contrast, 21-nucleotide small RNAs are associated with inverted repeats, including a novel microRNA family, and may function at the post-transcriptional level. Phytophthora infestans mycelium small RNAs were sequenced and aligned to the P. infestans genome for analysis.
Project description:Phytophthora infestans is most notorious oomycete causing a devastating disease on tomato called late blight. The molecular mechanisms involved in host-parasite interaction is still unexplored well. Investigation of changes in gene expression profile after pathogen infection to find out the mechanisms involved in infection process Second full expanded leaves from both healthy tomato plants (non-inoculated) and diseased tomato plants inoculated with Phytophthora infestans inoculum were used to extract total RNA for microarry analysis 12 hours post inoculation time.
Project description:In both plants and animals, alternative splicing (AS) increases transcriptome and proteome diversity, and play roles in developmental and immune responses. However, the extent of host genome-wide AS changes and how these changes are modulated by adaptive pathogens during disease remain largely unknown. Here we examined AS changes in Phytophthora infestans infected Solanum lycopersicum leaves using paired-end illumina RNA-seq. Quite a number of AS events are independent of gene differential expression, indicating that AS changes may be an additional layer of plant immunity against pathogens. we also demonstrated that P. infestans regulates plant immunity by repressing the AS of positive regulator of plant immunity or activating the AS of susceptibility factor. Furthermore, we established a splicing reporter system and screened some P. infestans splicing regulatory effectors (SREs) that are involved in plant mRNA alternative splicing process. In addition, we demonstrated that the WY motif of SRE3 is required for AS activity and virulence function. Overall, our current data reveal that P. infestans delivers SREs to disrupts host AS regulation machinery and subsequently suppress host immunity.