organism: Globodera pallida
description: Protocols: In vitro infection assays were performed as described by Goverse et al. (2000) with additions made by Zheng et al. (2022) and Schaveling et al., (2025). In brief, we used cuttings of the susceptible potato cultivar Desirée, which were grown on Gamborg B5 medium supplemented with either 20 g/L or 1.5 g/L of sucrose. After 14 days, cuttings were inoculated with 100 G. pallida Rookmaker J2s per plate.  We harvested infected roots at 1, 3, 6, and 9 days post inoculation (dpi). At 1 and 3 dpi, infected root segments of 15 plants were pooled per sample and at 6 and 9 dpi, infected root segments of 10 plants were pooled per sample.  Total RNA was extracted with the Maxwell 16 LEV-plant RNA kit (Promega, USA), according to the manufacturer's instructions. RNA samples were send for mRNA sequencing to BGI Genomics (Hong Kong, China). 1. Take a certain amount of total RNA samples, and use oligo dT beads to enrich mRNA with poly A tail; 2. mRNA molecules were fragmented into small pieces; 3. The fragmented mRNA was synthesized into first strand cDNA using random primers; 4. The second strand cDNA was synthesized with dUTP instead of dTTP; 5. The synthesized cDNA was subjected to end-repair and 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments; 6. Perform PCR amplification; 7. Library quality control; 8. Library circularization; 9. The library was amplified to make DNA nanoball (DNB); 10. Sequencing on DNBSEQ (DNBSEQ Technology) platform.
attributes.tag: ENA first public
attributes.value: 2025-11-25
attributes.tag: INSDC center name
attributes.value: Wageningen University and Research
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-16117:B_3.2_1,5
attributes.tag: age
attributes.value: 3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: collection date
attributes.value: not collected
attributes.tag: developmental stage
attributes.value: parasitic juvenile stage
attributes.tag: genotype
attributes.value: Potato variety Desiree
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: individual
attributes.value: Globodera pallida
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: infected root tissue
attributes.tag: replicate
attributes.value: 3
attributes.tag: sample name
attributes.value: E-MTAB-16117:B_3.2_1,5
attributes.tag: scientific_name
attributes.value: Globodera pallida
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: Rookmaker
attributes.tag: sucrose
attributes.value: 1.5