Plant growth Conditions

Plant growth Conditions 

For protoplast generation and collection of root slices, maize seeds were incubated in 3% (v/v) sodium hypochlorite for 8 min to sterilize the seeds. Then, seeds were then transferred to a sheet of wetted heavy-weight germination paper, rolled and covered with aluminum foil to prevent roots from exposure to direct light. Rolled paper was placed in a 2-gallon plastic container with at least 500ml of tap water and kept inside a Percival high light chamber for 7 days with a dark-light cycle of 16 hr light at 27ºC and 8hr dark at 24ºC and 50% humidity (also see https://currentprotocols.onlinelibrary.wiley.com/doi/abs/10.1002/cppb.20072). For plant propagation and crosses, maize plants were grown inside a greenhouse with controlled light and temperature under a 16 hr light at 28oC and 8hr dark at 22oC for 3 months. Setaria seeds were sterilized as described before and germinated on square plates containing 0.5X Murashige and Skoog (MS) medium. Plates were kept in a Percival high light chamber for 14 days with a dark-light cycle of 16 hr light at 27ºC and 8hr dark at 24ºC and 50% humidity until collection of root tissue for microscopic analyses. Setaria plants used for seed bulking and genetic crosses were grown in a walk-in chamber exposed to the same light/dark cycle and temperature as described above. All non-transgenic plants were Zea mays B73 background.

Sequencing platform

Library concentration was determined by quantitative PCR (qPCR) and sequenced with an Illumina NextSeq 550 platform using a 1x150 high-output configuration for replicate 1-3 (2 libraires per chips) or a Novaseq 6000 chip SP V2.5, 1x100 high-output for replicates 4-9 (4 libraries per chip)