ena-STUDY-JIC-10-04-2021-11:19:08:652-778
Sequencing of Phakopsora pachyrhizi isolate PpUFV02
The Phakopsora pachyrhizi PpUFV02 genome was sequenced with the PromethION platform of Oxford nanopore technology (Oxford, UK). The raw sequencing data of 110 Gbp was generated and was base-called using ONT Albacore 2.1. The resulting assembly was polished with the 50x Illumina PCR-free 150 bp PE reads.Four weeks old soybean plants were sprayed thoroughly on the abaxial surface of the leaves, and the plants were kept at 100% relative humidity in the dark for 24 h. After 24 h, plants were kept at 22°C and 16/8-h light/dark cycle. The leaf samples were collected from un-inoculated plants (0h) and infection-stages at 12, 24, 36, 72 and 168 HPI. Infection assay was performed in three biological replicates and three plants were used for each replicate. All the samples were stored in liquid Nitrogen after collection and kept in -80°C for further processing. Spores were harvested after 14 days post-inoculation and used for the RNA extraction. The urediniospores were germinated in vitro on the water surface in a square petri dish and kept for 6h at 24°C. The germinated-urediniospores were collected in a falcon tube and snap freeze in liquid nitrogen. The samples were freeze-dried and kept in -80°C for further processing.
The Phakopsora pachyrhizi PpUFV02 genome was sequenced with the PromethION platform of Oxford nanopore technology (Oxford, UK). The raw sequencing data of 110 Gbp was generated and was base-called using ONT Albacore 2.1. The resulting assembly was polished with the 50x Illumina PCR-free 150 bp PE reads.Four weeks old soybean plants were sprayed thoroughly on the abaxial surface of the leaves, and the plants were kept at 100% relative humidity in the dark for 24 h. After 24 h, plants were kept at 22°C and 16/8-h light/dark cycle. The leaf samples were collected from un-inoculated plants (0h) and infection-stages at 12, 24, 36, 72 and 168 HPI. Infection assay was performed in three biological replicates and three plants were used for each replicate. All the samples were stored in liquid Nitrogen after collection and kept in -80°C for further processing. Spores were harvested after 14 days post-inoculation and used for the RNA extraction. The urediniospores were germinated in vitro on the water surface in a square petri dish and kept for 6h at 24°C. The germinated-urediniospores were collected in a falcon tube and snap freeze in liquid nitrogen. The samples were freeze-dried and kept in -80°C for further processing.