イネ完熟種子糊粉層におけるアントシアニン生合成に係わる転写制御遺伝子のトランジェントアッセイ系

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  • A System for Transient Assay of Transcriptional Factors Controlling Anthocyanin Biosynthesis at the Aleurone Layer of Mature Rice Seed
  • イネ カンジュク シュシ コフンソウ ニ オケル アントシアニン セイゴウセイ ニ カカワル テンシャ セイギョ イデンシ ノ トランジュントアッセイケイ

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Various combinations of temperature and time for soaking seed were tested to establish a system for transient assay of transcriptional factors controlling anthocyanin biosynthesis to prove that the isolated gene is an inducible gene for anthocyanin expression at the aleurone layer of mature rice seed. The aleurone of mature seed of RIB-Pl with purple pericarp was sufficiently exposed under five combinations. Both Cl and B-Peru, transcriptional factors controlling anthocyanin biosynthesis of maize, were introduced by particle bombardment into the aleurone exposed under these five conditions. The highest expression of anthocyanin was obtained at the aleurone of the seeds soaked at 28°C for 4 hours. The effect of concentration of plasmids with Cl and B-Peru for introduction into the aleurone of A-58 mature seed with white pericarp soaked at 28°C for 4 hours was examined. Both Cl and B-Peru were found to give high expression of anthocyanin at the same concentration. Especially, 5 μg of Cl with 5 μg of B-Peru and 5 lig of Cl with 10 μg of B-Peru showed the highest expression. In conclusion, soaking of mature rice seed at 28°C for 4 hours and introduction of 5 lig of Cl with 51μg of B-Peru into the exposed aleurone was found to be a highly efficient system for transient assay of transcriptional factors controlling anthocyanin biosynthesis.

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