Project description:Investigation of whole genome gene expression level changes in a Lysobacter enzymogenes OH11 delta-clp mutant, compared to the wild-type strain.
Project description:ngs2013_07_pcapsici-effecaps-phytophthora capsici-The analysed RNAseq concerned the oomycete Phytophthora capsici in growth on pepper plants. 1/ How the adaptation of a pathogen to a host depends on its gene expression? 2/ How the plant host impacts the expression of pathogen genes at the very beginning of infection?-Two isolates of Phytophthora capsici were used: the Pc107 isolate (called A for adapted to pepper from INRAE GAFL Avignon), and the Pc273 isolate (N for non-adapted to pepper, collected on pumpkin in the USA). Two accessions of pepper (Capsicum annuum L., the host) were used: Yolo Wonder (YW, PM0031), susceptible (S) to Phytophthora capsici, and Criollo de Morelos 334 (CM334, PM0702), partially resistant (R). Inoculations were performed, as described in Lefebvre and Palloix (1996), by putting on the wounded stem a plug of mycelium. Inoculated plants were transferred to a growth chamber at 24°C/22°C temperature on a 12h/12h light/dark cycle. At 24 hours-post-inoculation, 12 total RNA samples were extracted from inoculated plants for the 4 host-isolate interactions: R_A, S_A, R_N and S_N. Each sample consisted of six pooled stem fragments. The stem fragments are the 5-mm region immediately under the visible stem necrosis. Samples were flash-frozen in liquid-nitrogen and stored at -80ºC. They were ground in liquid nitrogen with a cold mortar and pestle. Total RNA was extracted using QIAGEN Rneasy Plant Mini Kit. RNA-seq libraries were constructed at IPS2 POPS platform (France) by TruSeq_Stranded_mRNA_SamplePrep_Guide_15031047_D protocol (Illumina®, California, USA). Sequencing was conducted on an Illumina Hiseq2000 hosted by Genoscope (Evry, France). The RNA-seq samples have been sequenced in paired-end (PE) with a sizing of 260 bp and a read length of 100 bases, lane repartition and barcoding giving approximately 35 million of paired-end reads per sample.
