The subcellular localization of proteins is a fundamental aspect of protein functions. Determining the subcellular localization is important for understanding the biological functions of proteins. Here, we developed a set of rice organelle marker lines, in which the expressing fluorescent organelle markers could be used as comparative standards in determining the subcellular localization of the protein of interest. We constructed green fluorescent protein (GFP)- and/or Discosoma sp. red fluorescent protein (DsRed)-tagged organelle markers targeted to the endoplasmic reticulum (ER), mitochondria, Golgi apparatus, peroxisome, actin cytoskeleton, plastid, tonoplast, plasma membrane, and nucleus, respectively. The utility of the rice marker lines for protein subcellular localization studies was demonstrated by detecting a nucleus-localized OsWRKY45 and a mitochondria-associated NbHxk1 in protoplasts of the GFP-OsH2B and the ScCOX4-DsRed lines, respectively. Using a sheath-inoculation method, followed by a live-cell imaging, we detected co-localization of a Magnaporthe oryzae PWL2:mCherry : NLS fusion with the nucleus marker in the GFP-OsH2B rice epidermal cells, confirming the translocation of the M. oryzae effector PWL2 into host cells, and further demonstrating the feasibility of using the organelle marker lines for studying dynamics of proteins in rice cells in the interactions between rice and pathogens. The set of organelle marker lines developed in the present study, provides a valuable resource for protein subcellular localization studies in rice.

Reinforcement bar is the main skeleton of building materials. In order to increase the strength and plasticity of reinforcing bar, Nb and V elements are added to reinforcing bar to refine grain size. Fine grain strengthening can improve the strength of reinforcing bar and also improve the plasticity and toughness of reinforcing bar. The strain produced by the external force is very small in the grain interior and grain boundary of fine grains, and the difference of phase transformation is very small, the deformation is relatively uniform, and the chance of cracking caused by stress concentration is small. Tensile test and metallographic, scanning and energy spectrum analysis are carried out on the seismic steel bar. It is concluded that adding Nb and V elements will make the strength of the steel bar. And the plasticity is improved. 1 KEY WORDNb And V Elements; Refine Grain Size; Seismic Steel Bar.