1、第 43 卷第 2 期2023 年 4 月林 产 化 学 与 工 业Chemistry and Industry of Forest ProductsVol.43 No.2Apr.2023 收稿日期:2021-11-29 基金项目:国家自然科学基金资助项目(31300157);山西省自然科学基金项目(无编号);山西师范大学研究生科技创新项目(2020XSY025)作者简介:赵 猛(1980),男,山东济宁人,副教授,博士,研究方向为植物解剖学;E-mail:zhaomeng 。doi:10.3969/j.issn.0253-2417.2023.02.014Anatomical Structur
2、e of Gelsemium elegans Benth andthe Antibacterial Action of AlkaloidsZHAO MengZHAO Meng,LU Meiling,ZHAO Yating,WANG Yunxia,XUE Qiongqiong,WANG Xueqin(Life Science College,Shanxi Normal University,Taiyuan 030000,China)Abstract:In the present study,the microstructure of Gelsemium elegans vegetative or
3、gans was observed,and the alkaloids in plants were localized.Also,the antibacterial activity of the alkaloids was explored,and the antibacterial mechanism of the alkaloids was analyzed.The anatomical study of the structuralcharacteristics of G.elegans vegetative organs was carried out with anatomica
4、l methods,and through histochemical staining,thealkaloids in the plant were localized.Microscopic observation results showed that the structure of G.elegans performed the typicalcharacteristics of the dicotyledonous plants;the leaf of G.elegans was dorsiventral,and there were starch granules deposit
5、ed instem pith.The histochemical results showed that the vegetative organs of G.elegans contained alkaloids.Alkaloids were mainlydistributed in the mesophyll tissue of leaves.In the stem,alkaloids were mainly distributed in stem epidermis,cortex cells andphloem parenchyma cells.Also,alkaloids were d
6、istributed in parenchyma cells of pith and ray cells.Yellow-brown sedimentationalso could be observed in the cork cambium,the inner layer,the cortex,phloem parenchyma and ray cells of the rhizome.Theantibacterial activity results showed that the G.elegans alkaloid had antibacterial activity against
7、Listeria monocytogenesATCC19115,Staphylococcus aureus ATCC25923,Staphylococcus aureus CMCC26003,Escherichia coli ATCC25922,Escherichiacoli O157,Pseudomonas aeruginosa ATCC27853,and Pseudomonas aeruginosa CMCC10104.With the increase of alkaloidconcentration,the antibacterial effect was also enhanced.
8、The inhibitory effect on Listeria monocytogenes was the most obvious.After treatment with its minimum inhibitory and lethal concentrations,the morphology and internal structure of Staphylococcusaureus ATCC25923 and Escherichia coli O157 showed that both two kinds of bacteria changed in different deg
9、rees,such as roughsurface,depression,rupture of the cell membrane and cell wall,overflow of internal substance and even cell lysis.Themorphology of S.aureus changed at the minimum inhibitory concentration and the cell wall of E.coli was stratified.Thisindicated that G.elegans alkaloid destroyed the
10、permeability of the cell membrane,impeded the formation of the cell wall,andlysed the bacteria.The cell death at the minimum lethal concentration might be caused by the loss and cleavage of the mechanicalstrength and permeability of the wall.Key word:Gelsemium elegans Benth;anatomical structure;alka
11、loid distribution;bacteriostatic effect;bacteriostatic mechanismCLC number:TQ35 Document code:A Article ID:0253-2417(2023)02-0105-11引文格式:赵猛,路美玲,赵雅婷,等.钩吻解剖结构及钩吻生物碱抑菌作用研究(英文)J.林产化学与工业,2023,43(2):105-115.钩吻解剖结构及钩吻生物碱抑菌作用研究赵 猛,路美玲,赵雅婷,王云霞,薛琼琼,王雪琴(山西师范大学 生命科学学院,山西 太原 030000)摘 要:对钩吻营养和繁殖器官进行解剖学方面研究,并对植物体内
12、的生物碱的分布位置进行定位,此外,还探索了钩吻生物碱的抑菌活性,并对其抑菌机制进行分析研究。研究结果表明:钩吻结构符合一般双子叶植物的典型特征,叶为106 林 产 化 学 与 工 业第 43 卷异面叶,茎圆形,表皮有气孔,髓中有淀粉粒沉积。组织化学染色研究显示,钩吻全株含有生物碱,生物碱类物质主要分布在叶的叶肉组织中,尤其栅栏组织含量较多,维管束中少或无;在茎中,生物碱类物质主要分布在茎的表皮、皮层细胞、韧皮薄壁细胞中,另外,在茎髓周围的薄壁细胞及髓射线细胞中也分布有生物碱类物质;在钩吻根状茎的木栓形成层、栓内层、皮层、韧皮部的薄壁细胞及射线细胞中均可观察到。通过滤纸片法、二倍微量稀释法测定钩
13、吻生物碱的抑菌活性和最小抑菌浓度,结果显示,钩吻生物碱具有广谱抗菌性,对李斯特氏菌 ATCC19115、金黄色葡萄球菌 ATCC25923、金黄色葡萄球菌 CMCC26003、大肠杆菌 ATCC25922、大肠杆菌 O157、铜绿假单胞菌 ATCC27853、铜绿假单胞菌 CMCC10104 这7 种供试菌都具有抑菌活性,且随着生物碱浓度的增加,抑菌效果也随之加强,其中对李斯特菌抑菌效果最明显。选择革兰氏阳性菌金黄色葡萄球菌 ATCC25923 和革兰氏阴性菌大肠杆菌 O157,测定出它们的最小致死质量浓度(MBC)分别为 500 和 750 g/L,利用透射电子显微镜观察钩吻生物碱在其最小抑
14、菌浓度和最小致死浓度处理前后金黄色葡萄球菌ATCC25923 和大肠杆菌 O157 菌体形态及内部结构的变化,结果发现 2 种菌体都发生了不同程度的改变,具体表现在菌体表面粗糙,出现凹陷,细胞膜和细胞壁发生破裂,内容物溢出甚至引发细胞裂解,发现在最小抑菌浓度下金黄色葡萄球菌形态发生改变,而大肠杆菌的细胞壁也出现分层现象,这表明生物碱破坏了菌体细胞壁的形成,因为细菌形状受到肽聚糖囊泡影响。在最小致死浓度下细胞死亡可能是由于壁的机械强度和渗透性的丧失并发生裂解所引起的。关键词:钩吻;解剖结构;生物碱分布;抑菌作用;抑菌机制Gelsemium elegans is evergreen vine of
15、 Gelsemium.Gelsemium was first attributed to the Apocynaceae.Later,according to the relationship between the number of chromosomes and phylogeny,it was revised to beattributed to Loganiaceae,and then it was separately formed into a family,the Gelsemiaceae1-3.G.elegansis highly toxic,and its main act
16、ive ingredient is steroidal alkaloid,which can significantly inhibit centralnervous system activity4-6.There are three species of Gelsemium plants in the world,including G.elegans inChina and G.sempervirens,G.rankinii in North America7-8.China has a long history of the application ofG.elegans,but it is mainly for external use,such as eliminating swelling4.At present,lots of studies havebeen conducted on the chemical composition,toxicology and pharmacological effects of G.elegans.Thestudies indic
