IC4R011-Epigenomic-2015-11001567

Project Title

• Epigenomic modification in rice controls meiotic recombination and segregation distortion

Plant Culture & Treatment

• F1 plants were obtained by crosses between wildtype NB or asDDM1 (female) and KS (male), and grown in a greenhouse. The effects of DDM1 knockdown and TSA treatment on chromatin modification were confirmed by Southern blot analysis with a probe for centromeric repeats (Supplementary Fig. S1b and c). Four F1 plants for each sample [F1 (NB 9 KS),TSA-treated F1 (NB 9 KS) and F1 (asDDM1 9 KS)]were grown in a greenhouse and self-pollinated. Five panicles were taken from each F1 plant, and 15 seeds per panicle were sown in soil. A total of 190 plants[(three F1 plants 9 5 panicles 9 10 seeds) ? (one F1 plant 9 4 panicles 9 10 seeds) = 190] were selected randomly, and genomic DNA was extracted. Genotyping of the F2 progenies was done by PCR using primers shown in Supplementary Table S3. Germination rates were at least 91.3 % for F2 (NB 9 KS),93.6 % for TSA-treated F2 (NB 9 KS) and 93.6 % for F2 (asDDM1 9 KS). The frequencies of morphologically aberrant plants (albino, striped leaf and pale green leaf) were 1.3 % for F1 (NB 9 KS), 0.7 % for TSA-treated F1 (NB 9 KS) and 3.0 % for F1(asDDM1 9 KS). These plants were not used for further analysis.
• F1 seeds were sterilized with 2.5 % sodium hypochlorite for 20 min, washed three times with sterile water and imbibed in sterile water or 100 lM TSA (Sigma)at 30 �C overnight. Imbibed seeds were washed three times with sterile water and grown on Murashige–

Skoog medium/0.4 % agar at 30 �C for 7–10 days without TSA. Seedlings were then transferred to soil in a greenhouse and grown further to obtain self-pollinated seeds.

Labs working on this Project

• Agrogenomics Research Center, National Institute of Agrobiological Sciences, Kannondai 2-1-2,Tsukuba 305-8602, Japan
• Institute of Society for Techno-Innovation of Agriculture,Forestry, and Fisheries, Tsukuba 305-0854, Japan
• Institute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, Japan

Corresponding Author

• Yoshiki Habu:habu@affrc.go.jp