钩吻解剖结构及钩吻生物碱抑菌作用研究

doi:10.3969/j.issn.0253-2417.2023.02.014

摘要/Abstract

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

Abstract:

In the present study, the microstructure of Gelsemium elegans vegetative organs 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 structural characteristics of G. elegans vegetative organs was carried out with anatomical methods, and through histochemical staining, the alkaloids in the plant were localized. Microscopic observation results showed that the structure of G. elegans performed the typical characteristics of the dicotyledonous plants; the leaf of G. elegans was dorsiventral, and there were starch granules deposited in stem pith. The histochemical results showed that the vegetative organs of G. elegans contained alkaloids. Alkaloids were mainly distributed in the mesophyll tissue of leaves. In the stem, alkaloids were mainly distributed in stem epidermis, cortex cells and phloem parenchyma cells. Also, alkaloids were distributed in parenchyma cells of pith and ray cells. Yellow-brown sedimentation also could be observed in the cork cambium, the inner layer, the cortex, phloem parenchyma and ray cells of the rhizome. The antibacterial activity results showed that the G. elegans alkaloid had antibacterial activity against Listeria monocytogenes ATCC19115, Staphylococcus aureus ATCC25923, Staphylococcus aureus CMCC26003, Escherichia coli ATCC25922, Escherichia coli O157, Pseudomonas aeruginosa ATCC27853, and Pseudomonas aeruginosa CMCC10104. With the increase of alkaloid concentration, the antibacterial effect was also enhanced. 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 Staphylococcus aureus ATCC25923 and Escherichia coli O157 showed that both two kinds of bacteria changed in different degrees, such as rough surface, depression, rupture of the cell membrane and cell wall, overflow of internal substance and even cell lysis. The morphology of S. aureus changed at the minimum inhibitory concentration and the cell wall of E. coli was stratified. This indicated that G. elegans alkaloid destroyed the permeability of the cell membrane, impeded the formation of the cell wall, and lysed the bacteria. The cell death at the minimum lethal concentration might be caused by the loss and cleavage of the mechanical strength and permeability of the wall.

Key words: Gelsemium elegans Benth, anatomical structure, alkaloid distribution, bacteriostatic effect, bacteriostatic mechanism

中图分类号:

TQ35

赵猛, 路美玲, 赵雅婷, 王云霞, 薛琼琼, 王雪琴. 钩吻解剖结构及钩吻生物碱抑菌作用研究[J]. 林产化学与工业, 2023, 43(2): 105-115.

Meng ZHAO, Meiling LU, Yating ZHAO, Yunxia WANG, Qiongqiong XUE, Xueqin WANG. Anatomical Structure of Gelsemium elegans Benth and the Antibacterial Action of Alkaloids[J]. Chemistry and Industry of Forest Products, 2023, 43(2): 105-115.

图/表 7

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Table 1

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strain	inhibition zone diameter of G.elegans alkaloids at different concn./mm
strain	500 g/L	250 g/L	125 g/L	62.5 g/L
P. aeruginosa ATCC27853	11.00±0.23	10.00±0.23	9.00±0.00	7.00±0.00
P. aeruginosa CMCC10104	12.50±0.05	11.00±0.25	9.50±0.21	7.50±0.00
E. coli ATCC25922	13.20±0.00	13.00±0.05	11.00±0.00	7.50±0.25
E. coli O157	13.50±0.35	13.00±0.25	12.00±0.00	8.50±0.00
S. aureus ATCC25923	13.50±0.35	13.50±0.00	12.00±0.25	8.00±0.00
S. aureus CMCC26003	13.80±0.35	13.50±0.35	12.50±0.00	8.00±0.25
L.monocytogenes ATCC19115	15.00±0.05	14.00±0.21	13.00±0.12	9.00±0.26

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