RNA-seq analysis of Rhizophagus sp. strain HR1 under P-deficient condition
The frozen mycelia (20 to 40 mg FW) obtained at time zero and 4 h after Pi application were ground with a metal-cone in a 3-ml O-ring sealed plastic tube (Yasui Kikai, Osaka, Japan) using Multi-Beads Shocker (Yasui Kikai) at 2,000 rpm for 4 ? 10 s, during which the samples were cooled in liquid nitrogen at each interval. Total RNA was extracted with RNeasy Plant Mini Kit (Qiagen), followed by RNase-free DNase I (Qiagen) treatment. Sequencing libraries (n = 3 for each time point) were constructed using 500 ng of total RNA with TruSeq RNA Sample Prep kit (Illumina) following the manufacturer?s instructions.