中国长白山乌头属5种植物表观结构植纹鉴定研究
摘要
本文采用植纹专利技术,对长白山地区66科98属248种植物成熟叶片的植纹特征进行研究,将它们在光学显微镜下的叶表皮特征归纳为15种植纹类型,即无规则型、不定型、大小不等型、平列型、横列型、十字交叉型、平列不等型、辐射型、围绕型、双耳型、交叉不规则型、保卫细胞参差不等型、单子叶植物具长短细胞的哑铃型、单子叶植物不具长短细胞的肾型和单子叶植物具长短细胞的肾型。结果表明:结合植纹类型、叶片表皮细胞垂周壁类型以及表皮附属物类型可以对长白山地区药用植物进行有效的鉴别。
随后运用统计学原理,选取长白山地区6种珍稀药用植物(高山红景天(Rhodiola sachalinensis A. Ber.),黄芩(Scutellaria baicalensis Georgi.),射干(Belamcanda Chinesis (L. ) DC.),长白棘豆(Oxytropis anertii Nakai ex Kitag.),独角莲(Typhonium giganteum Engl.)和防风(Saposhnikovia divaricata (Turcz.) Schischk.))以及乌头属5种植物(北乌头(Aconitum kusnezoffii Reichb.),草地乌头(Aconitum umbrosum (Korsh.) Kom.),黄花乌头(Aconitum coreanum (Lévl.) Rapaics),吉林乌头(Aconitum kirinense Nakai)和细叶黄乌头(Aconitum barbatum Pers.))为研究对象,测量其各项植纹特征参数并进行统计分析。实验结果表明,从3个层次(植纹特征,量化后各植纹特征的平均值,各植纹特征间特有的显著相关性和最优的线性回归方程)应用植纹技术进行鉴别,具有极高的准确性和可信性。
对上述乌头属5种植物茎和叶的解剖学研究结果表明:北乌头、草地乌头和细叶黄乌头的茎中维管束为拟散生维管束,即不具有束中形成层,具有厚壁组织所围成的鞘状结构;韧皮纤维外侧具有环状分泌道结构;所有维管束排列成内外两轮,星散分布在茎的基本组织当中。而吉林乌头和黄花乌头的茎中维管束为环状排列的外韧无限维管束,具有束中形成层,没有厚壁鞘和分泌组织结构。另外,北乌头的叶片为不具有栅栏组织的等面叶类型;草地乌头和细叶黄乌头叶片的栅栏组织分化不明显。这3种植物的叶片内具有环状分泌道结构,并且分泌道紧邻输导组织和平脉叶肉细胞。黄花乌头叶片的通气组织非常发达,叶肉细胞彼此几乎不联系;吉林乌头的叶片具有非常典型的栅栏组织柱状细胞。此2种乌头属植物不具有环状分泌道结构。
研究还发现,5种乌头属植物都具有适应湿生环境的解剖结构特征,即茎的表皮细胞外切向壁角质层和机械组织不发达;叶肉通气组织发达且彼此连接成网状;叶片表皮含有大量的异形胞,角质层不发达;有的种类叶片下表皮气孔向外凸出等。实验结果证实,植物的结构与其所处的环境是相适应的。
此外,本文运用数值分类学原理,选用66个包括植物宏观、解剖以及植纹特征在内的各类性状,对上述5种乌头属植物进行聚类分析。结果表明,草地乌头和细叶黄乌头具有非常近的亲缘关系,而同为牛扁亚属牛扁组的吉林乌头与它们具有明显的差异,因此建议将吉林乌头单列一系。相比之下,北乌头与草地乌头和细叶黄乌头这类植物有着较近的亲缘关系。
针对乌头属药用植物中多糖成分研究的不足,以乌头属最著名的药用植物川乌(乌头Aconitum carmichaeli Debx.的主根),附子(乌头Aconitum carmichaeli Debx.的侧根)和草乌(北乌头Aconitum Kusnezoffii Reichb.的主根)为实验材料,对其中的多糖成分进行含量、组成以及生物学活性研究。结果表明,3种乌头多糖AWP(川乌),LWP(附子)和KWP(草乌),除了产率上的差别以外,糖含量和单糖组成基本一致,都主要由葡萄糖组成,说明这是一类葡聚糖。
抗肿瘤和免疫调节作用测定结果表明,高低剂量的3种乌头多糖不仅能够显著抑制小鼠体内肿瘤生长,还能够拮抗化疗药物环磷酰胺(Cy)对机体免疫功能的抑制,提高机体的非特异性免疫,细胞和体液免疫,且都存在一致的剂量依赖关系,因而,推测乌头多糖是通过提高机体免疫能力来抑制体内肿瘤生长的。
乌头多糖组成和药理学活性研究表明,无论相同乌头属药用植物的主根和侧根,还是不同植物的相同药用部位,所含有的乌头多糖都是组成和活性非常相近的一类活性多糖,是除乌头碱以外另一种重要的活性物质,作为抗肿瘤药物和免疫调节剂具有广阔的应用前景。
In the present study, the abaxial leaf epidermises of 66 families, 98 genera, 248 species medicinal plants in Changbai Mountain were observed with optical microscopy using the Plant Print technology. The characters of the Plant Print were summarized to be 15 types, including anomocytic, indefinite, anisocytic, paracytic, laterocytic, staurocytic, inequipolar, actinocytic, cyclocytic, auritate, decussate irregular, guard cell disparate, dumbbell-shape guard cell monocotyledon with long short epidermis cells, reniform guard cell monocotyledon and reniform guard cell monocotyledon with long short epidermis cells. The results indicated that medicinal plants in Changbai Mountain could be effectively identified according to the types of Plant Print characters, anticlinal wall and appendages of the leaf epidermises.
The Plant Print characters (the length, width, perimeter and area of guard cells, as well as the perimeter and area of subsidiary cells, stomatal index and density) of 6 rare and precious medicinal plants (including Rhodiola sachalinensis A. Ber., Scutellaria baicalensis Georgi., Belamcanda Chinesis (L.) DC., Oxytropis anertii Nakai ex Kitag., Typhonium giganteum Engl. and Saposhnikovia divaricata (Turcz.) Schischk.), which belonging to different families and genus were measured and statistically analyzed, as well as 5 species of Aconitum plants, including A. kusnezoffii Reichb., A.umbrosum (Korsh.) Kom., A. coreanum (Lévl.) Rapaics, A. kirinense Nakai and A. barbatum Pers. The results indicated that based on 3 levels of characters, including Plant Print characteristics, the means of quantitated Plant Print characteristics, and significant correlations and optimal equations of linear regression between the characteristics, the identification of plants by Plant Print technology and statistics analysis had extremely high accuracy and reliability, no matter the plants without any relationships in family and genus or difficult to be discriminated in macroscopy taxonomy.
The stems and leave of the 5 Aconitum species were applied to do the anatomical studies using optical microscopic technique. The result of anatomical study on stems showed that vasculars of stems of A. kusnezoffii, A.umbrosum and A. barbatum were quasi atactosteles, which did not have fascicular cambium, but contained bundle sheath composed of sclerenchyma. The stems had annular secretory canals outside of phloem fibres. All the vasculars arrayed two rings and dispersively distributed in elementary tissue of stem; vasculars of stems of A. coreanum and A. kirinense were open bundles with intradascicular cambium, but without bundle sheath and bundle sheath.
The result of anatomical study on leave showed that leave of A. kusnezoffii were isobilateral leave without palisade tissue; palisade tissue of A.umbrosum and A. barbatum did not obviously differentiated. The above 3 species had annular secretory canals, and their mesophyll cells scarcely connected with each other. Aerating tissue of leave of A. coreanum was well developed; leave of A. coreanum had typical palisade tissue. A. coreanum and A. kirinense did not have annular secretory canals.
Moreover, the anatomical structures of stems and leave of 5 Aconitum species had wet habitat-structural characters: mechanical tissue and corneous layer in tangential wall of epidermal cell of stems were less developed; aerating tissue of diachyma was developed, and air cavity connected to be reticulated; epidermis of leave had large amount of heterocyst, corneous layer was less developed; stoma in abaxial leaf epidermises of some species were outer projectured. It is confirmed that structure and habitat are unified.
The 5 Aconitum species were applied to cluster analysis with 11 macroscopy binary characters of plants according to numerical taxonomy theory. The result of numerical taxonomy was consistent with that of traditional taxonomy, indicating the numerical taxonomy method used in the present study had high reliability. So we chose 66 characters, including morphologic characteristics, anatomical characteristics of aerial parts nutritive organ and Plant Print characteristics to do cluster analysis. The result indicated that A. umbrosum and A. barbatum had very close genetic relationships, but A. kirinense, which also belongs to Subgen. Praconitum, Sect. Praconitum, exhibited obvious difference from A. umbrosum and A. barbatum. While A. kusnezoffii showed close genetic relationships with the group of A. umbrosum and A. barbatum. Therefore, morphologic characteristics, anatomical characteristics of aerial parts nutritive organ and Plant Print characteristics were quantized and calculated to generate the quantitative comparison between the 5 Aconitum species according to the mathematic model, which is a scientific, objective and precise taxonomy compared to traditional taxonomy.
Radix Aconiti (“Chuanwu”, the mother roots of A. carmichaeli Debx.), Radix Aconiti Lateralis (“Fuzi”, the daughter roots of A. carmichaeli) and Radix Aconiti Kusnezoffii (“Caowu”, the mother roots of A. Kusnezoffii) are the most commonly used traditional Chinese medicines (TCM) of Aconitum species. In the present study, the water-soluble polysaccharides of Radix Aconiti, Radix Aconiti Lateralis and Radix Aconiti Kusnezoffii, were extracted, and named as AWP, LWP and KWP, respectively. The chemical analyses indicated the water-soluble polysaccharides of the three materials were all mainly composed of glucose, implying they were glucans.
The antitumor and immunomodulatory assays showed that AWP, LWP and KWP not only had good antitumor activities, but also could enhance the host’s immunity including nonspecific immunity, cellular immunity and humoral immunity, and restore the antitumor drug-suppressed immune function.
The composition and pharmacological activity exhibited polysaccharides from Aconitum species, whatever from mother roots or daughter roots of the same plant, or mother roots of different plants, was a sort of bioactive substances. Therefore, the polysaccharides from Aconitum species might be conveniently exploited to be good immune stimulating modifiers and had great potential to be applied in the tumor therapy.
随后运用统计学原理,选取长白山地区6种珍稀药用植物(高山红景天(Rhodiola sachalinensis A. Ber.),黄芩(Scutellaria baicalensis Georgi.),射干(Belamcanda Chinesis (L. ) DC.),长白棘豆(Oxytropis anertii Nakai ex Kitag.),独角莲(Typhonium giganteum Engl.)和防风(Saposhnikovia divaricata (Turcz.) Schischk.))以及乌头属5种植物(北乌头(Aconitum kusnezoffii Reichb.),草地乌头(Aconitum umbrosum (Korsh.) Kom.),黄花乌头(Aconitum coreanum (Lévl.) Rapaics),吉林乌头(Aconitum kirinense Nakai)和细叶黄乌头(Aconitum barbatum Pers.))为研究对象,测量其各项植纹特征参数并进行统计分析。实验结果表明,从3个层次(植纹特征,量化后各植纹特征的平均值,各植纹特征间特有的显著相关性和最优的线性回归方程)应用植纹技术进行鉴别,具有极高的准确性和可信性。
对上述乌头属5种植物茎和叶的解剖学研究结果表明:北乌头、草地乌头和细叶黄乌头的茎中维管束为拟散生维管束,即不具有束中形成层,具有厚壁组织所围成的鞘状结构;韧皮纤维外侧具有环状分泌道结构;所有维管束排列成内外两轮,星散分布在茎的基本组织当中。而吉林乌头和黄花乌头的茎中维管束为环状排列的外韧无限维管束,具有束中形成层,没有厚壁鞘和分泌组织结构。另外,北乌头的叶片为不具有栅栏组织的等面叶类型;草地乌头和细叶黄乌头叶片的栅栏组织分化不明显。这3种植物的叶片内具有环状分泌道结构,并且分泌道紧邻输导组织和平脉叶肉细胞。黄花乌头叶片的通气组织非常发达,叶肉细胞彼此几乎不联系;吉林乌头的叶片具有非常典型的栅栏组织柱状细胞。此2种乌头属植物不具有环状分泌道结构。
研究还发现,5种乌头属植物都具有适应湿生环境的解剖结构特征,即茎的表皮细胞外切向壁角质层和机械组织不发达;叶肉通气组织发达且彼此连接成网状;叶片表皮含有大量的异形胞,角质层不发达;有的种类叶片下表皮气孔向外凸出等。实验结果证实,植物的结构与其所处的环境是相适应的。
此外,本文运用数值分类学原理,选用66个包括植物宏观、解剖以及植纹特征在内的各类性状,对上述5种乌头属植物进行聚类分析。结果表明,草地乌头和细叶黄乌头具有非常近的亲缘关系,而同为牛扁亚属牛扁组的吉林乌头与它们具有明显的差异,因此建议将吉林乌头单列一系。相比之下,北乌头与草地乌头和细叶黄乌头这类植物有着较近的亲缘关系。
针对乌头属药用植物中多糖成分研究的不足,以乌头属最著名的药用植物川乌(乌头Aconitum carmichaeli Debx.的主根),附子(乌头Aconitum carmichaeli Debx.的侧根)和草乌(北乌头Aconitum Kusnezoffii Reichb.的主根)为实验材料,对其中的多糖成分进行含量、组成以及生物学活性研究。结果表明,3种乌头多糖AWP(川乌),LWP(附子)和KWP(草乌),除了产率上的差别以外,糖含量和单糖组成基本一致,都主要由葡萄糖组成,说明这是一类葡聚糖。
抗肿瘤和免疫调节作用测定结果表明,高低剂量的3种乌头多糖不仅能够显著抑制小鼠体内肿瘤生长,还能够拮抗化疗药物环磷酰胺(Cy)对机体免疫功能的抑制,提高机体的非特异性免疫,细胞和体液免疫,且都存在一致的剂量依赖关系,因而,推测乌头多糖是通过提高机体免疫能力来抑制体内肿瘤生长的。
乌头多糖组成和药理学活性研究表明,无论相同乌头属药用植物的主根和侧根,还是不同植物的相同药用部位,所含有的乌头多糖都是组成和活性非常相近的一类活性多糖,是除乌头碱以外另一种重要的活性物质,作为抗肿瘤药物和免疫调节剂具有广阔的应用前景。
In the present study, the abaxial leaf epidermises of 66 families, 98 genera, 248 species medicinal plants in Changbai Mountain were observed with optical microscopy using the Plant Print technology. The characters of the Plant Print were summarized to be 15 types, including anomocytic, indefinite, anisocytic, paracytic, laterocytic, staurocytic, inequipolar, actinocytic, cyclocytic, auritate, decussate irregular, guard cell disparate, dumbbell-shape guard cell monocotyledon with long short epidermis cells, reniform guard cell monocotyledon and reniform guard cell monocotyledon with long short epidermis cells. The results indicated that medicinal plants in Changbai Mountain could be effectively identified according to the types of Plant Print characters, anticlinal wall and appendages of the leaf epidermises.
The Plant Print characters (the length, width, perimeter and area of guard cells, as well as the perimeter and area of subsidiary cells, stomatal index and density) of 6 rare and precious medicinal plants (including Rhodiola sachalinensis A. Ber., Scutellaria baicalensis Georgi., Belamcanda Chinesis (L.) DC., Oxytropis anertii Nakai ex Kitag., Typhonium giganteum Engl. and Saposhnikovia divaricata (Turcz.) Schischk.), which belonging to different families and genus were measured and statistically analyzed, as well as 5 species of Aconitum plants, including A. kusnezoffii Reichb., A.umbrosum (Korsh.) Kom., A. coreanum (Lévl.) Rapaics, A. kirinense Nakai and A. barbatum Pers. The results indicated that based on 3 levels of characters, including Plant Print characteristics, the means of quantitated Plant Print characteristics, and significant correlations and optimal equations of linear regression between the characteristics, the identification of plants by Plant Print technology and statistics analysis had extremely high accuracy and reliability, no matter the plants without any relationships in family and genus or difficult to be discriminated in macroscopy taxonomy.
The stems and leave of the 5 Aconitum species were applied to do the anatomical studies using optical microscopic technique. The result of anatomical study on stems showed that vasculars of stems of A. kusnezoffii, A.umbrosum and A. barbatum were quasi atactosteles, which did not have fascicular cambium, but contained bundle sheath composed of sclerenchyma. The stems had annular secretory canals outside of phloem fibres. All the vasculars arrayed two rings and dispersively distributed in elementary tissue of stem; vasculars of stems of A. coreanum and A. kirinense were open bundles with intradascicular cambium, but without bundle sheath and bundle sheath.
The result of anatomical study on leave showed that leave of A. kusnezoffii were isobilateral leave without palisade tissue; palisade tissue of A.umbrosum and A. barbatum did not obviously differentiated. The above 3 species had annular secretory canals, and their mesophyll cells scarcely connected with each other. Aerating tissue of leave of A. coreanum was well developed; leave of A. coreanum had typical palisade tissue. A. coreanum and A. kirinense did not have annular secretory canals.
Moreover, the anatomical structures of stems and leave of 5 Aconitum species had wet habitat-structural characters: mechanical tissue and corneous layer in tangential wall of epidermal cell of stems were less developed; aerating tissue of diachyma was developed, and air cavity connected to be reticulated; epidermis of leave had large amount of heterocyst, corneous layer was less developed; stoma in abaxial leaf epidermises of some species were outer projectured. It is confirmed that structure and habitat are unified.
The 5 Aconitum species were applied to cluster analysis with 11 macroscopy binary characters of plants according to numerical taxonomy theory. The result of numerical taxonomy was consistent with that of traditional taxonomy, indicating the numerical taxonomy method used in the present study had high reliability. So we chose 66 characters, including morphologic characteristics, anatomical characteristics of aerial parts nutritive organ and Plant Print characteristics to do cluster analysis. The result indicated that A. umbrosum and A. barbatum had very close genetic relationships, but A. kirinense, which also belongs to Subgen. Praconitum, Sect. Praconitum, exhibited obvious difference from A. umbrosum and A. barbatum. While A. kusnezoffii showed close genetic relationships with the group of A. umbrosum and A. barbatum. Therefore, morphologic characteristics, anatomical characteristics of aerial parts nutritive organ and Plant Print characteristics were quantized and calculated to generate the quantitative comparison between the 5 Aconitum species according to the mathematic model, which is a scientific, objective and precise taxonomy compared to traditional taxonomy.
Radix Aconiti (“Chuanwu”, the mother roots of A. carmichaeli Debx.), Radix Aconiti Lateralis (“Fuzi”, the daughter roots of A. carmichaeli) and Radix Aconiti Kusnezoffii (“Caowu”, the mother roots of A. Kusnezoffii) are the most commonly used traditional Chinese medicines (TCM) of Aconitum species. In the present study, the water-soluble polysaccharides of Radix Aconiti, Radix Aconiti Lateralis and Radix Aconiti Kusnezoffii, were extracted, and named as AWP, LWP and KWP, respectively. The chemical analyses indicated the water-soluble polysaccharides of the three materials were all mainly composed of glucose, implying they were glucans.
The antitumor and immunomodulatory assays showed that AWP, LWP and KWP not only had good antitumor activities, but also could enhance the host’s immunity including nonspecific immunity, cellular immunity and humoral immunity, and restore the antitumor drug-suppressed immune function.
The composition and pharmacological activity exhibited polysaccharides from Aconitum species, whatever from mother roots or daughter roots of the same plant, or mother roots of different plants, was a sort of bioactive substances. Therefore, the polysaccharides from Aconitum species might be conveniently exploited to be good immune stimulating modifiers and had great potential to be applied in the tumor therapy.
引文
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