利用荧光显微镜识别近缘物种--以柿树属(Genus Persicaria Mill)为例

A. Gudkova, A. S. Chistyakova, A. I. Slivkin, A. S. Bolgov
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One of such selective methods for diagnosing plant materials, along with the well-known and used in pharmaceutical analysis, is luminescence microscopy.Aim. The goal was to study the features of the luminescence of closely related species of the genus Persicaria Mill. to improve the process of their identification and standardization of official types of raw materials.Materials and methods. For work, 10 species of the genus Persicaria Mill. were used from families Polygonaceae Juss., most common in the Central Chernozem region: P. maculosa Gray, P. tomentósa Bicknell, P. lapathifólia Delarbre, P. nodósa Opiz, P. scábra Moldenke, P. brittingeri Opiz, P. hydropiper Delarbre, P. minor Opiz, P. amphibia Delarbre, P. amphibia var. terrestris Munshi & Javeid. The plants were cut at a height of about 15 cm from the soil during mass flowering in the summer of 2020 in the Voronezh region, dried in the shade. 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引用次数: 0

摘要

导言。药用植物材料的标准化和质量评估问题是当前最重要的问题之一。为此,需要对一系列参数进行评估,其中最主要的是对所分析植物对象真实性的评估。柿属(Persicaria Mill.)植物的典型问题是在存在近缘物种的情况下如何确定正式物种,其中只有两个物种被用于医疗实践,而且这些植物的药材都有规范性文件。可用于医疗用途的植物物种与杂质的形态和解剖特征相似,因此需要使用更多的现代分析方法。发光显微镜是诊断植物材料的选择性方法之一,也是众所周知的药物分析方法。目的是研究密切相关的柿属植物的发光特征,以改进其鉴定过程和官方原料类型的标准化。工作中使用了蓼科 Juss.中的 10 个柿属物种,它们在切尔诺泽姆中部地区最为常见:P. maculosa Gray、P. tomentósa Bicknell、P. lapathifólia Delarbre、P. nodósa Opiz、P. scábra Moldenke、P. brittingeri Opiz、P. hydropiper Delarbre、P. minor Opiz、P. amphibia Delarbre、P. amphibia var. terrestris Munshi & Javeid。2020 年夏天,在沃罗涅日地区大量开花时,在植株距土壤约 15 厘米处剪下,在阴凉处晾干。在研究之前,植物被分成不同的形态部分(茎、叶、叶柄、花铃、花、果),为此使用了标本馆的干燥样本,并在玻璃载玻片上拉直后对各个部分进行了检查。为了评估物体的发光情况,使用了带有发光喷嘴外壳的 Micromed-3 Lum 显微镜(俄罗斯)。结果表明,大毛状体、传导系统、气孔的保卫细胞、细胞壁的增厚以及某些物种的花托和腺体内容物的强烈荧光是所有研究物种的共同特征。所研究物种的组织发光有许多特征:P. hydropiper 的所有形态部位都有许多明亮发光的花托和腺体。对于 P. maculosa 而言,首次确定了沿叶脉存在大型花托。brittingeri 的花被、腺体和花铃丝状毛清晰可见,并发出黄绿色的荧光。Tomentósa 的叶腺体与其他物种不同,不发光。P. scábra 和 P. tomentósa 的花被呈黄橙色。P. lapathifólia的膜状毛状体清晰可见,呈褐色。在 P. amphibia var. terrestris 中,数量最多的长束毛状体发出明亮的绿色光芒。首次将发光显微镜方法应用于识别参数的分析和鉴定,建立了新的诊断特征并将其可视化,从而能够最准确地鉴定柿属物种。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Luminescence Microscopy in the Identification of Closely related Species Using the Example of the Genus Persicaria Mill.
Introduction. The issue of standardization and quality assessment of medicinal plant materials is currently one of the most important. To do this, a number of parameters are evaluated, the primary of which is the assessment of the authenticity of the analyzed plant object. The problem of identifying official species in the presence of closely related species is typical for plants belonging to the genus Persicaria Mill., among which only two species are used in medical practice and there is regulatory documentation for the herbs of these plants. The similarity of morphological and anatomical features of plant species acceptable for medical use and impurities interprets the need to use additional modern methods of analysis. One of such selective methods for diagnosing plant materials, along with the well-known and used in pharmaceutical analysis, is luminescence microscopy.Aim. The goal was to study the features of the luminescence of closely related species of the genus Persicaria Mill. to improve the process of their identification and standardization of official types of raw materials.Materials and methods. For work, 10 species of the genus Persicaria Mill. were used from families Polygonaceae Juss., most common in the Central Chernozem region: P. maculosa Gray, P. tomentósa Bicknell, P. lapathifólia Delarbre, P. nodósa Opiz, P. scábra Moldenke, P. brittingeri Opiz, P. hydropiper Delarbre, P. minor Opiz, P. amphibia Delarbre, P. amphibia var. terrestris Munshi & Javeid. The plants were cut at a height of about 15 cm from the soil during mass flowering in the summer of 2020 in the Voronezh region, dried in the shade. Previously, before the study, the plants were divided into morphological components (stems, leaves, petioles, bells, flowers, fruits), for which herbarium samples were used in dried form and individual parts were examined, straightened on a glass slide. To assess the luminescence of objects, a Micromed-3 Lum microscope (Russia) with a luminescent nozzle housing was used.Results and discussion. It was revealed that the intense fluorescence of large trichomes, the conducting system, guard cells of stomata, thickening of cell walls, and, in some species, the contents of receptacles and glands, is common to all studied species. A number of characteristic features of tissue luminescence for the studied species have been revealed: P. hydropiper has numerous brightly luminous receptacles and glands on all morphological parts. For P. maculosa, the presence of large receptacles along the veins was established for the first time. The perianth, glandules, and filiform hairs of the bell of P. brittingeri are clearly visible and have a yellowish-green luminescence. The leaf glands of P. tomentósa, unlike other species, do not have luminescence. The perianths of P. scábra and P. tomentósa have a yellowish-orange glow. The membranous trichomes are clearly visible in P. lapathifólia and stand out with a brownish tint. The largest number of long tufted trichomes with a bright greenish glow is characteristic of P. amphibia var. terrestris.Conclusion. The method of luminescence microscopy was first applied to the analysis and identification of identification parameters, new diagnostic features were established and visualized, allowing the most accurate identification of species of the genus Persicaria.
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