Haider G, Kislinger T, Kutchan T M
Laboratorium für Molekulare Biologie, Universität München, Germany.
Biochem Biophys Res Commun. 1997 Dec 18;241(2):606-10. doi: 10.1006/bbrc.1997.7842.
Four barbiturates, barbituric acid, butethal, phenobarbital, and 2-thiobarbituric acid, of fourteen tested were found to induce accumulation of benzophenanthridine alkaloids in cell suspension cultures of the California poppy Eschscholzia california. When the plant cell suspension cultures were treated with 1 mM barbiturate, alkaloids accumulated to 100 mg/l within four days. This is a level comparable to that achieved with 300 microM concentration of the established secondary metabolite inducer methyl jasmonate. In contrast to methyl jasmonate, barbituric acid, and 2-thiobarbituric acid, butethal and phenobarbital treatment resulted in a different alkaloid profile, suggesting that only select cytochrome P-450 genes were activated by these latter two barbiturates. RNA gel blot analysis of barbiturate induced cell cultures confirmed that transcripts of at least two benzophenanthridine alkaloid biosynthetic genes cyp80b1 (encoding the cytochrome P-450-dependent monooxygenase (S)-N-methylcoclaurine 3'-hydroxylase) and bbe1 (encoding the covalently flavinylated berberine bridge enzyme) increased up to 5- to 7-fold over control values.
在对14种受试物质进行检测时发现，四种巴比妥酸盐，即巴比妥酸、布他比妥、苯巴比妥和2-硫代巴比妥酸，可诱导加州罂粟（Eschscholzia california）细胞悬浮培养物中苯并菲啶生物碱的积累。当植物细胞悬浮培养物用1 mM巴比妥酸盐处理时，生物碱在四天内积累至100 mg/l。这一水平与用300 microM浓度的已确定的次生代谢物诱导剂茉莉酸甲酯所达到的水平相当。与茉莉酸甲酯、巴比妥酸和2-硫代巴比妥酸不同，布他比妥和苯巴比妥处理导致了不同的生物碱谱，这表明只有特定的细胞色素P-450基因被后两种巴比妥酸盐激活。对巴比妥酸盐诱导的细胞培养物进行RNA凝胶印迹分析证实，至少两个苯并菲啶生物碱生物合成基因cyp80b1（编码细胞色素P-450依赖性单加氧酶（S）-N-甲基coclaurine 3'-羟化酶）和bbe1（编码共价黄素化的小檗碱桥酶）的转录本比对照值增加了5至7倍。
