Naik Tanushree, Vanitha Shanadrahalli Chandrashekaraiah, Rajvanshi Pradumn Kumar, Chandrika Manjegowda, Kamalraj Subban, Jayabaskaran Chelliah
Department of Biochemistry, Indian Institute of Science, Bangalore, 560 012, India.
Curr Microbiol. 2018 Feb;75(2):206-212. doi: 10.1007/s00284-017-1367-y. Epub 2017 Oct 24.
Eighteen endophytic fungi were isolated from various tissues of Datura metel and genes encoding for putrescine N-methyltransferase (PMT), tropinone reductase 1 (TR1) and hyoscyamine 6β-hydroxylase (H6H) were used as molecular markers for PCR-based screening approach for tropane alkaloids (TAs) producing endophytic fungi. These fungi were identified taxonomically by sequence analysis of the internal transcribed spacer region (ITS1-5.8S-ITS2) and also based on morphological characteristics of the fungal spore as Colletotrichum boninense, Phomopsis sp., Fusarium solani, Colletotrichum incarnatum, Colletotrichum siamense and Colletotrichum gloeosporioides. The production of TAs hyoscyamine and scopolamine by the fungi has been ascertained using chromatography and spectroscopy methods by comparison with the standards. Among the fungi, the highest yields of hyoscyamine (3.9 mg/L) and scopolamine (4.1 mg/L) were found in C. incarnatum culture. This is the first report of endophytic fungi possess the PMT, TR1 and H6H genes and produces TAs. These endophytic fungi have significant potential to be applied in fermentation technology to meet the demands for TAs economically.
从白花曼陀罗的不同组织中分离出18种内生真菌，并将编码腐胺N-甲基转移酶（PMT）、托品酮还原酶1（TR1）和莨菪碱6β-羟化酶（H6H）的基因用作基于PCR的筛选方法的分子标记，以筛选产生托烷生物碱（TA）的内生真菌。通过对内部转录间隔区（ITS1-5.8S-ITS2）的序列分析，并根据真菌孢子的形态特征，将这些真菌分类鉴定为博宁炭疽菌、拟茎点霉属、茄病镰刀菌、肉色炭疽菌、暹罗炭疽菌和胶孢炭疽菌。通过与标准品比较，采用色谱和光谱方法确定了真菌产生的TA莨菪碱和东莨菪碱。在这些真菌中，肉色炭疽菌培养物中莨菪碱（3.9 mg/L）和东莨菪碱（4.1 mg/L）的产量最高。这是关于内生真菌拥有PMT、TR1和H6H基因并产生TA的首次报道。这些内生真菌在发酵技术中具有巨大的应用潜力，能够经济地满足对TA的需求。
