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糖酸还原酶在氧化苦参碱内生真菌中的潜在作用。

Potential role for saccharopine reductase in swainsonine metabolism in endophytic fungus, Undifilum oxytropis.

机构信息

Molecular Biology Program, New Mexico State University, Las Cruces, NM 88003, USA.

出版信息

Fungal Biol. 2012 Aug;116(8):902-9. doi: 10.1016/j.funbio.2012.05.007. Epub 2012 Jun 16.

DOI:10.1016/j.funbio.2012.05.007
PMID:22862918
Abstract

Locoweed plants in the southwestern United States often harbour a slow-growing endophytic fungus, Undifilum oxytropis (Phylum: Ascomycota; Order: Pleosporales), which produces a toxic alkaloid, swainsonine. Consumption of U. oxytropis by grazing animals induces a neurological disorder called locoism for which the toxic alkaloid swainsonine has been reported to be the causal agent. Little is known about the biosynthetic pathway of swainsonine in endophytic fungi, but previous studies on non-endophytic ascomycetous fungi indicate that pipecolic acid and saccharopine are key intermediates. We have used degenerate primers, Rapid amplification of cDNA ends (RACE)-PCR and inverse PCR to identify the gene sequence of U. oxytropis saccharopine reductase. To investigate the role of this gene product in swainsonine metabolism, we have developed a gene deletion system for this slow-growing endophyte based on our recently established transformation protocol. A strain of U. oxytropis lacking saccharopine reductase had decreased levels of saccharopine and lysine along with increased accumulation of pipecolic acid and swainsonine. Thus, saccharopine reductase influences the accumulation of swainsonine and its precursor, pipecolic acid, in U. oxytropis.

摘要

美国西南部的疯草植物常寄生一种生长缓慢的内生真菌,即 Oxytropis unifila(真菌界:子囊菌门;目:腔孢目),其能产生一种有毒生物碱,即苦马豆素。食草动物食用 Oxytropis unifila 会引发一种神经紊乱疾病,称为疯草病,而苦马豆素已被报道为这种疾病的致病因子。关于内生真菌中苦马豆素的生物合成途径知之甚少,但先前对非内生子囊菌的研究表明,吡咯啉酸和蔗糖碱是关键的中间产物。我们使用简并引物、RACE-PCR 和反向 PCR 来鉴定 Oxytropis unifila 蔗糖碱还原酶的基因序列。为了研究该基因产物在苦马豆素代谢中的作用,我们根据最近建立的转化方案,为这种生长缓慢的内生真菌开发了一种基因缺失系统。缺乏蔗糖碱还原酶的 Oxytropis unifila 菌株的蔗糖碱和赖氨酸水平降低,同时吡咯啉酸和苦马豆素的积累增加。因此,蔗糖碱还原酶影响 Oxytropis unifila 中苦马豆素及其前体吡咯啉酸的积累。

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