T-field image (H). (I ) A protoplast cell co-expressing OsAP65 FP (I
T-field image (H). (I ) A protoplast cell co-expressing OsAP65 FP (I) and a PVC marker RFP tVSR2 (J), a merged image (K), along with a bright-field image (L). Scale bars=10 m. (This figure is accessible in colour at JXB online.)needed for pollen germination and pollen tube development. When OsAP65 was disrupted, this substrate might not be degraded inside a timely manner, resulting in impaired pollen germination and pollen tube growth. However, the physiological function of OsAP65 won’t be entirely clear until its substrates are identified. A recent post showed that two rice AP genes, OsAP25 and OsAP37, that were promoted by ETERNAL TAPETUM 1, trigged programmed cell death in tapetal cells in rice anthers (Niu et al., 2013). OsAP65 might take part in a molecular pathway causing male sterility inside the exact same way as OsAP25 and OsAP37. Nevertheless, the present final results demonstrate a crucial function for OsAP65 in fertilization through its function in pollen tube development, but not pollen maturation.AcknowledgementsWe thank Dr Gynheung An (POSTECH, Korea) for providing the mutants, Dr Liwen Jiang (The Chinese University of Hong Kong, Hong Kong, China) for offering the PVC marker plasmid RFP tVSR2 and the Golgi marker plasmid Man1 FP, and Dr Jian Xu (Huazhong Agricultural University, China) for supplying the the mitochondrial marker plasmid F1-ATPase-:RFP. This work was supported by grants from the National 863 Project (2012AA10A303) and the National All-natural Science Foundation of China (30921091 and 31201190).5-HT6 Receptor site References Supplementary dataSupplementary data are obtainable at JXB on the web. Figure S1. Characterization on the OsAP65 T-DNA insertion line. Figure S2. PCR final results for genotyping the progeny of OsAP65+/plants. Figure S3. Features of OsAP65 protein. Figure S4. Schematic diagrams in the OsAP65 gene and complementation vector. Figure S5. Genetic analyses and genotyping in the T1 generation from OsAP65 transformation plants. Table S1. Primers for PCR evaluation. Table S2. Detailed facts of rice tissues in Fig. 5A.Asakura T, Watanabe H, Abe K, Arai S. 1995. Rice aspartic proteinase, oryzasin, HDAC1 Gene ID expressed during seed ripening and germination, has a gene organization distinct from these of animal and microbial aspartic proteinases. European Journal of Biochemistry 232, 773. Bi X, Khush GS, Bennett J. 2005. The rice nucellin gene ortholog OsAsp1 encodes an active aspartic protease devoid of a plant-specific insert and is strongly expressed in early embryo. Plant and Cell Physiology 46, 878. Chen J, Ouyang Y, Wang L, Xie W, Zhang Q. 2009. Aspartic proteases gene family members in rice: gene structure and expression, predicted protein functions and phylogenetic relation. Gene 442, 10818. Chen J, Ding J, Ouyang Y, et al. 2008. A triallelic technique of S5 is usually a big regulator in the reproductive barrier and compatibility ofA rice aspartic protease regulates pollen tube growth |indica aponica hybrids in rice. Proceedings from the National Academy of Sciences, USA 105, 114361441. Dai X, You C, Chen G, Li X, Zhang Q, Wu C. 2011. OsBC1L4 encodes a COBRA-like protein that affects cellulose synthesis in rice. Plant Molecular Biology 75, 33345. Davies DR. 1990. The structure and function from the aspartic proteinases. Annual Review of Biophysics and Biophysical Chemistry 19, 18915. de Graaf BHJ, Cheung AY, Andreyeva T, Levasseur K, Kieliszewski M, Wu H-m. 2005. Rab11 GTPase-regulated membrane trafficking is crucial for tip-focused pollen tube growth in tobacco. The Plant Cell 17, 256457.