Ca2+ sensor-mediated ROS scavenging suppresses rice immunity and is exploited by a fungal effector
This dataset contains all original data for the disease resistance phenotypes in wild-type, rod1, ROD1-OE, rod1 complemented with Ca2+ binding mutants, RIP1 CRISPR mutants, RIP1-OE, APIP6 CRISPR mutants, APIP6-OE, ROD1-SNP, and rod1 complemented with AvrPiz-t. This dataset is related to: https://doi.org/10.1016/j.cell.2021.09.009.
Steps to reproduce
Blast fungal inoculation was conducted as previously described (Deng et al., 2017; Zhai et al., 2019). Briefly, M. oryzae (isolate TH12) was cultured on complete agar medium at 25 C for 10 days to produce spores. Leaves at tillering stage were punch-inoculated with spores diluted in distilled H2O at a concentration of 100000 spores/mL. For blast inoculation in seedlings, two-week-old plants were sprayed with blast spore suspensions. Inoculated seedlings were kept in a dew growth chamber at 26 C for 24 h in darkness, and then grown at 26 C with 12 h/12 h (day/night) and 90% relative humidity for 7 days. Lesion lengths were measured and fungal growth was determined using quantitative PCR (Kawano et al., 2010) to measure the amount of Pot2 transposon DNA, which was normalized to the rice ubiquitin gene (LOC_Os03 g13170). Sheath blight inoculation was carried out at tillering stages in the paddy field as previously reported with slight modification (Yin et al., 2018). Briefly, Rhizoctonia solani AG1-IA (isolate RH-9) was cultured on PDA (potato-dextrose-agar) plates for 2 d at 28 C. Short (0.8-1.0 cm) wooden toothpicks were sterilized and co-incubated with fungal plugs for 3 d at 28_C. The fungi growing toothpicks were then inserted into the third leaf sheath. Sheath blight symptom was recorded at 7 dpi with more than 20 sheaths each genotype (2 sheaths per plant) analyzed. For the Xoo infection assay, bacterial strain PXO99A was grown on a peptone sucrose agar (PSA) medium at 28_C for 3 days. The bacteria were collected and then suspended in sterilized water at a concentration of OD600 = 1.0, which were used to infect two month-old plants (tillering stage) by the leaf-clipping method (Gao et al., 2017). Lesion length was measured at 14 dpi. All fungal and bacterial infections were repeated independently at least three times.