Os06g0643300

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The combination of RISBZ1 and RPBF play critical roles in global processes, including SSP gene expression, during seed development.

Annotated Information

Function

1.combinatorial interactions between RISBZ1 and RPBF in the transcriptional apparatus confer a synergistic effect on many rice SSP genes, encoding glutelins, prolamins andaglobulin, an observation that points to the potential of rice seed for extremely high SSP gene expression.

2. To examine the effect of RISBZ1 and RPBF on seed quality, we measured the total protein, starch and total lipid contents. As observed on SDS-PAGE analysis (Figure 2a), the protein concentrations were lower in KD lines.

3.In wild-type seeds, the surfaces of the endosperm cross sections were crystallized, with a glossy sheen (Figure 4i).

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Expression

1.In wild-type seeds, the accumulation of RISBZ1 was very high until 14 days after flowering (DAF), then radically declined to low or undetectable levels (Figure 1c). RPBF accumulation was detectable throughout the endosperm development, and reached a maximum at 14 DAF (Figure 1c).

2. In wild-type seed,RISBZ1 was highly expressed until 7 and 14 DAF (Figure 1d).RPBF expression reached a maximum at 14 DAF, declining afterwards to approximately the same level as found at 7 DAF (Figure 1d).

3. In both KD-RISBZ1 and KD-RPBF seeds, glutelin GluA was slightly decreased, and the level of GluB was comparable with that of the wild type. The GluD level was enhanced in KD-RISBZ1 and completely lost in KD-RPBF.(figure2)

Figure x1.jpg


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Localization

RISBZ1 and RPBF are localized in the nucleus, and interact in rice protoplasts. When immunofluorescently detected with anti-HA and 4¢,6-diamidino-2-phenylindole (DAPI) staining, it was shown that bothRISBZ1andRPBF were localized in the nucleus. (Figure 5a)


Figure x5a.jpg

Mutation

1.In KDRISBZ1 seed,RISBZ1expression was much greater at 7 DAF, but was subsequently lower than that of wild-type seed (Figure 1d). RPBFexpression was slightly higher at 7 DAF, but was lower at 14 and 21 DAF than in wild-type seed (Figure 1d).RISBZ1expression was similar in KD-RPBF and KD-RISBZ1 seeds (Figure 1d).

2.The seed phenotypes of KD-RISBZ1, KD-RPBF and KD-RISBZ1/KD-RPBF were opaque and wrinkled (Figure 4a–h), with KD-RISBZ1 being the least affected, and KD-RISBZ1/ KD-RPBF being severely affected (Figure 4a–h).


Figure x4a-h.jpg

Labs working on this gene

1.Transgenic Crop Research and Development Centre, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan.

2 Graduate School of Science and Technology, Chiba University, Matsudo, Chiba 271-8510, Japan

References

1.Taiji Kawakatsu , Masayuki P. Yamamoto, Satoru M. Touno et al(2009). Compensation and interaction between RISBZ1 and RPBF during grain filling in rice, The Plant Journal(2009)59, 908–920.

2. Adachi, T., Izumi, H., Yamada, T., Tanaka, K., Takeuchi, S., Nakamura, R. and Matsuda, T.(1993) Gene structure and expression of rice seed allergenic proteins belonging to the alphaamylase/trypsin inhibitor family.Plant Mol. Biol.21, 239–248.

Structured Information