Additional file 7: Figure S4. of The role of Tre6P and SnRK1 in maize early kernel development and events leading to stress-induced kernel abortion

Maize kernel culture. Kernels were prepared by hand excision of the center one-third of 3 DAP ears (A), and removal of kernels and a small portion of cob tissue (B), maintaining sufficient cob tissue to permit nutrients to be taken up by the immature kernel. Immature kernels were sterilized and grown in sucrose rich (150 mM Sucrose, 1% MS agar), or sucrose deficient media (1% MS agar) on petri dishes with equal spacing to ensure that there was equal competition for nutrients (C). Embryos were kept at 24 °C in the dark for 48 h, then transferred to plates with sucrose for another 48 h. Replicate samples are removed every 8–12 h, kernels were removed from cob tissue (D & E), weighed, and immediately frozen in liquid N2. (XLSX 310 kb)