The Initiation Switch for the Seed Formation in Rice

Methods

Summary

1.      Cloning the genes of interest with the inducible promoters into the specific vector pTF101.1 (offered by Dr. Kan Wang, Iowa State University), respectively, and then transfer these constructs into Agrobacteria.

2.      Transform rice callus with the Agrobacteria and culture the transformed callus into seedling plants (to be carried out by Iowa State University).

3.      Grow the transformed rice seedling in growth chamber and identify the expected mutant plants.

4.      Observe the fluorescent signals from rice egg cells and zygotes at different stages using the fluorescent microscope, so as to characterize the level change of KRP and Fb3 proteins and their interaction in the initiation of rice seed formation.

Challenges

The transformation in rice requires very complicated facilities and professional skills which are not available at our campus. That is why we have to cooperate with Dr. Kan Wang, Iowa State University with the world-recognized best service in rice transformation. However, it is too expensive for us.

Pre Analysis Plan

Once the above stated signals are viewed, it will be the first convincing evidence that KRP5  protein is the substrate of Fb3 at the beginning of post-zygotic cell division, indicating this crucial cell cycle process is regulated by Fb3 protein in rice seed formation. In case that the observation result is not as expected, we may have to turn to transient transformation in mass isolated rice egg cells and zygotes with the same constructs or considering other candidate F-box and KRP proteins and their interactions.