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对有毒和无毒品种的L.愈伤组织提取物进行化学分析

Chemical analysis of callus extracts from toxic and non-toxic varieties of L.

作者信息

Leyva-Padrón Gerardo, Vanegas-Espinoza Pablo Emilio, Evangelista-Lozano Silvia, Del Villar-Martínez Alma Angélica, Bazaldúa Crescencio

机构信息

Centro de Desarrollo de Productos Bióticos, Instituto Politécnico Nacional, Yautepec, Morelos, México.

出版信息

PeerJ. 2020 Nov 11;8:e10172. doi: 10.7717/peerj.10172. eCollection 2020.

Abstract

L. belongs to Euphorbiaceae family, and it synthesizes flavonoid and diterpene compounds that have showed antioxidant, anti-inflammatory, anticancer, antiviral, antimicrobial, antifungal and insecticide activity. Seeds of this plant accumulate phorbol esters, which are tigliane type diterpenes, reported as toxic and, depending on its concentration, toxic and non-toxic varieties has been identified. The aim of this work was to characterize the chemical profile of the extracts from seeds, leaves and callus of both varieties (toxic and non-toxic) of , to verify the presence of important compounds in dedifferentiated cells and consider the possibility of using these cultures for the massive production of metabolites. Callus induction was obtained using NAA (1.5 mg L) and BAP (1.5 mg L) after 21 d for both varieties. Thin layer chromatography analysis showed differences in compounds accumulation in callus from non-toxic variety throughout the time of culture, diterpenes showed an increase along the time, in contrast with flavonoids which decreased. Based on the results obtained through microQTOF-QII spectrometer it is suggested a higher accumulation of phorbol esters, derived from 12-deoxy-16-hydroxy-phorbol ( 365 [M+H]), in callus of 38 d than those of 14 d culture, from both varieties. Unlike flavonoids accumulation, the MS chromatograms analysis allowed to suggest lower accumulation of flavonoids as the culture time progresses, in callus from both varieties. The presence of six glycosylated flavonoids is also suggested in leaf and callus extracts derived from both varieties (toxic and non-toxic), including: apigenin 6---L-arabinopyranosyl-8- -D-xylopyranoside ( 535 [M+H]), apigenin 4'--rhamnoside ( 417 [M+H]), vitexin ( 433 [M+H]), vitexin 4'--glucoside-2″--rhamnoside ( 741 [M+H]), vicenin-2 ( 595 [M+H]), and vicenin-2,6″--glucoside ( 757 [M+H]).

摘要

L. 属于大戟科,它能合成黄酮类和二萜类化合物,这些化合物已显示出抗氧化、抗炎、抗癌、抗病毒、抗菌、抗真菌和杀虫活性。这种植物的种子积累佛波酯,它们是替告皂苷元型二萜,据报道有毒,并且根据其浓度,已鉴定出有毒和无毒品种。这项工作的目的是表征这两个品种(有毒和无毒)的种子、叶子和愈伤组织提取物的化学特征,验证去分化细胞中重要化合物的存在,并考虑使用这些培养物大量生产代谢物的可能性。两个品种在21天后使用萘乙酸(1.5毫克/升)和苄氨基嘌呤(1.5毫克/升)诱导出愈伤组织。薄层色谱分析表明,在整个培养期间,无毒品种愈伤组织中化合物的积累存在差异,二萜类化合物随时间增加,而黄酮类化合物则减少。基于通过微型四极杆飞行时间串联质谱仪获得的结果,表明在38天的愈伤组织中,来自12-脱氧-16-羟基佛波醇(365 [M+H])的佛波酯积累量高于14天培养的愈伤组织,两个品种均如此。与黄酮类化合物的积累不同,质谱色谱图分析表明,随着培养时间的延长,两个品种愈伤组织中黄酮类化合物的积累量较低。在来自两个品种(有毒和无毒)的叶子和愈伤组织提取物中也表明存在六种糖基化黄酮类化合物,包括:芹菜素6---L-阿拉伯吡喃糖基-8- -D-木吡喃糖苷(535 [M+H])、芹菜素4'- -鼠李糖苷(417 [M+H])、牡荆素(433 [M+H])、牡荆素4'- -葡萄糖苷-2″--鼠李糖苷(741 [M+H])、异荭草苷(595 [M+H])和异荭草苷-2,6″--葡萄糖苷(757 [M+H])。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a7b6/7666564/ca459f051351/peerj-08-10172-g001.jpg

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