79. イネのサクラネチン生合成酵素OsNOMTの単離・同定(口頭発表,植物化学調節学会第46回大会)

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Sakuranetin, a flavonoid phytoalexin in rice, is biosynthesized from naringenin by naringenin 7-O-methyltransferase, NOMT. The NOMT activity is induced prior to sakuranetin accumulation, and antimicrobial activity of naringenin against M oryzae is enhanced by the methylation. These observations suggest that regulation mechanism of NOMT is one of the key steps for rice defense responses. However rice NOMT has not been identified. Previously, NOMT was partially purified from UV-treated wild-type rice leaves, but the major component in the purified fraction showed caffeic acid O-methyltransferase (OsCOMT) activity but not NOMT activity, suggesting that the rice NOMT (OsNOMT) is present as a minor component in the purified fraction. We therefore attempted to isolate OsNOMT using an OsCOMT1 mutant, oscomt1. Crude protein extracted from UV treated oscomt1 leaves was sequentially applied to ammonium sulfate precipitation, anion exchange purification and affinity purification using adenosine agarose. The NOMT activity in the partially purified sample was concentrated 400-fold compared with the starting sample. 2 OMTs, 0s04g0175900 and Os12g0240900, were identified by MALDI-TOF/TOF analysis from 40kDa protein on SDS-PAGE in the sample. We revealed that the recombinant protein of Os12g0240900 but not Os04g0175900 has NOMT activity. Furthermore Os12g0240900 expression was induced by JA treatment in rice leaves. With these results, we concluded that Os12g0240900 functions as OsNOMT.
2011-10-03

79. イネのサクラネチン生合成酵素OsNOMTの単離・同定(口頭発表,植物化学調節学会第46回大会)

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