IC4R012-RNA-Seq-2012-22347394

Project Title

Transcriptomic Analysis of Rice (Oryza sativa) Developing Embryos Using the RNA-Seq Technique

Rice (O. sativa L.ssp. indica cv.9311) plants were grown in a greenhouse at Wuhan University (30u330N, 114u190E), China. Some spikelets were tagged at the initiation of pollination, and harvested at 3–5, 7, and 14 DAP. Before 3 DAP, the zygote reiterates cell divisions to form a globular embryo with no apparent morphological differentiation. At 5 DAP, the first leaf primordium is visible on the opposite side of the coleoptile, and the shoot apical meristem becomes dome-shaped at 7 DAP [32]. Morphological maturity of the rice embryo was noted at approximately 14 DAP. Since the number of embryos at 3 DAP was extremely low, a mixture of embryos at 3–5 DAP was collected as one sample. The three-staged embryos were taken from spikelets with microdissection needles under a dissection microscope (Olympus, Tokyo, Japan) according to the manual microdissection method and frozen immediately in liquid nitrogen for total RNA extraction. Three embryo clusters were collected from 15 different plants at 3–5, 7, and 14 DAP during the growing season. The embryos were randomly selected from each cluster and pooled with embryos from other plants at the same greenhouse site, resulting in three independent pools for each developmental stage.

State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan, China