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)玉米籽粒培养体系。实验材料制备流程：手动切取授粉后3天（days after pollination，DAP）果穗的中央三分之一区段（图A），剥离籽粒并保留少量穗轴组织（图B），确保留存足够的穗轴组织以向未成熟籽粒供应养分。将所得未成熟籽粒进行灭菌处理，随后分别接种于高蔗糖培养基（150 mM蔗糖、1% MS琼脂培养基（Murashige and Skoog agar medium））或缺蔗糖培养基（仅含1% MS琼脂培养基）的培养皿中，籽粒间保持均匀间距以确保养分竞争条件一致（图C）。将培养体系置于24 ℃黑暗环境中培养48小时，随后转移至含蔗糖的培养基中继续培养48小时。每间隔8~12小时收取重复样本：将籽粒从穗轴组织上剥离（图D、E），称量鲜重后立即置于液氮（liquid nitrogen，N₂）中速冻保存。（XLSX格式，310 KB）