First data on the phytoliths of some medicinal plants of Primorye region, Russian Federation.
Phytoliths (Greek, phyton--plant and lithos stone) --micrometer silicon particles are synthesized in plants have long been the subject of considerable scrutiny. First, the term "fitolitaries" was introduced by the German scientist, a foreign member of the Russian Academy of Sciences, the founder of micropaleontology Christian Gottfried Ehrenberg, who established their biogenic origin . A detailed study of silicon microparticles in plants was started only a century later. In Russia, the first studies on phytoliths were initiated by soil scientists .
By now, questions on phytoliths systematic are developed quite well, mainly of grasses [3-6]. It was found that the amount of accumulation in plants phytoliths depends mainly on the taxonomic affiliation and age of the plant . Differences within taxons are determined by several environmental factors, primarily by soil geochemistry . By now, experts suggested three basic versions of the functions of phytoliths in plants: structural, physiological, and protective [7, 8].
Structural function allows the plant to maintain the required density, thus preserve upright vertical position. Physiological function of phytoliths may manifest itself in that the silicon dioxide is involved in the neutralization of the excess of certain chemical elements such as aluminum, manganese, falling within the plants together with soil moisture, and reducing their toxic effects. The protective function of phytoliths allows the plant to withstand both pathogenic fungi and insects on the one hand and eating with the other herbivores [6, 7]. The method of phytoliths analysis has long been used for reconstruction of paleovegetation, paleolandscapes, and archaeology [4, 9, 10].
The present study is the first attempt to describe the phytoliths of the modern flora of Primorye. Earlier phytoliths in vegetation of Far East were hardly studied. The possibility of correlation of the medicinal properties of plants and morphotypes of phytoliths is of particular interest.
Materials and Methods
For the study, we chose nine species of medicinal plants and one type of fungus growing in the Primorsky region (Table 1).
Plants were collected in 2013 in Terneisky area (near the Sikhote-Alin and township Terni) and Anuchinsky area (near the village Anuchino) of Primorye.
Extraction of phytoliths was conducted according to the procedure of Piperno : The leaf samples were burned for 2 h in a muffle furnace at 450[degrees]C, and then cooled at room temperature. Thereafter washed with 10% solution of HCl, and then with concentrated nitric acid and distilled water. The studies were conducted using the hardware-software complex, which includes a dedicated computer, microscope AxioScope A1 (Zeiss, Germany), a scanner and a digital camera AxioCam 3. For statistics 150 phytoliths in each plant were morfometred. Definition of morphotypes of phytoliths was conducted by International Code for Phytolith Nomenclature 1.0 .
Results and Discussion
Berberis amurensis (Berberidaceae)
Widely distributed within the Far East, it is also found in Primorye and the Amur region. Alkaloid berberine was isolated from its leaves and bark. Tincture of the plants causes vasoconstriction, several increases blood clotting, strengthens contractions of the uterus. It is applied in obstetric practice to stop atonic postpartum hemorrhage associated with inflammatory diseases .
"Star-shaped" morphotypes of phytoliths were found in the leaves of Berberis, which are not found in other types of test plants (Figure 1) and their maximum amount with respect to other morphotypes is (48%). Phytoliths such as "irregular ruminate" (Figure 2) amounted to about 17%, "long cage"--14% and "smooth cylinders"--3%. Solitary met spherical shape, in the form of "short cells", and "cylinder", "box", and phytoliths of "epidermal hair cells". Morphometric parameters of phytoliths in Amur barberry are shown in the Table 2.
Schisandra chinensis (Schisandraceae)
The only species of this family, which is found on the territory of Russia (common in Primorsky and Khabarovsk territories, Southern Sakhalin, and on Shikotan, Kunashir, and other Kuril Islands). Tincture of Schisandra chinensis (Chinese magnolia vine) is used as a stimulant for the prevention of fatigue under heavy exertion .
To investigate, this plant's phytoliths were collected its different parts: vine, leaves, and berries. The greatest number and variety of morphotypes of phytoliths was found out in the leaves, vine in phytoliths met in the least (Figures 3-6).
In vine, the most common phytoliths are "irregular ruminate" (hereinafter given names of morphotypes of phytoliths, according to International Code for Phytolith Nomenclature 1.0. --about 43%, "a sleek curved cylinder"-10%, and "short elongated cell"--7%. Singular encounter "short lobed cells", "short truncated cells", "smooth spheroids", "ellipsoids smooth", and "wrinkled plate".
In berries of Schisandra chinensis, "irregular smooth" phytoliths are dominated (about 55%). "Ellipsoid wrinkled" was found in 7%, other forms were similar to phytoliths of the vine.
Phytoliths in leaves were similar to phytoliths of berries and vines: 40% of them are "irregular ruminate", 8%--in the form of the "ellipsoid wrinkled", 7%--"parallelepiped elongated smooth".
Isolated phytoliths in leaves are more diverse than in the fruit and vine: it is "an elongated parallelepiped grungy", "smooth thinned parallele-piped with rounded edges", and "wrinkled spherical shape". Morphometric parameters of phytoliths from Schisandra chinensis are given in Table 3.
Rosa acicularis (Rosaceae)
In the Far East grows 11 kinds of wild rose, but only 5 of them are considered to be well studied. Accepted criterion of the value of wild roses as a source of medicinal raw materials is the presence of large amounts of ascorbic acid.
The main form of phytoliths in rose fruit is (Figure 7) "irregular ruminate" (about 34%); Further descending "ellipsoid wrinkled" (18%), "grungy parallelepiped thinned with rounded edges" (11%), "short elongated cell" (10%), "smooth cylinder" (9%). Singular encounter: "short saddle-shaped cell", "cell short truncated cone-shaped", "cylinder wrinkled", "smooth curved cylinder". Morphometric parameters of phytoliths in Rosa acicularis are given in Table 4.
Bergenia pacifica (Saxifragaceae)
Widespread in the Primorye and the Amur region on the outskirts of screes, on the upper edge of the forest or among shrubs. Usually forms thickets. Bergenia pacifica is very close to growth in Siberia leather bergenia and is overlooking a replacement for the Far East. The pharmacological study of drugs of leaves of leather bergenia found their ability to narrowing of blood vessels and compact vascular wall. It was also revealed their antimicrobial and anti-inflammatory action .
In the leaves of Bergenia pacifica was found the largest amount as compared with other plant species of phytoliths of rounded shapes (Figure 8): "spherical"--about 20%, as "ellipsoid"--about 13%, and "parallelepipeds with rounded edges--about 9%". Most phytoliths was "irreg-ular smooth"--about 37%. Solitary met two cylinder forms (flat and curved) in the form of "parallelepiped", as well as the "short elongated cells". Morphometric parameters of phytoliths from Bergenia pacifica are shown in Table 5.
Eleutherococcus senticosus (Araliaceae) Distribution area of eleutherococcus includes Primorye, Amur, and Sakhalin. Eleutherococcus has a strong stimulating effect. Reception of extract of this plant leads to a significant increase in mental and physical performance. Eleutherococcus' important property is its beneficial effect on the body's resistance to many pathogenic factors .
To study this type of phytoliths only leaves were collected. The vast majority of leaves found in phytoliths are irregular ruminate (Figure 9) and irregular smooth (Figure 10)--a total of 85%. Other forms were met in small amounts, "the subordinate epidermal the hair", "wrinkled cylinder", "smooth curved cylinder", "smooth ellipsoid". Morphometric parameters of phytoliths from Eleutherococcus senticosus are shown in Table 6.
Phellodendron amurense (Rutaceae)
Widespread in the Primorye and southern regions of the Amur region, it is used as a tonic, antiseptic, antipyretic, and styptic. In folk medicine of Nanai, fresh Amur cork is used for the treatment of chronic dermatitis and tinea. In China, ointments and powders from best Amur cork are successfully used for some forms of eczema. The leaves of velvet contain carotene--to 3.3 mg%, vitamin C (about 22 mg%) and E--12 mg% .
In fruits of Amur cork 60% were found phytoliths of "irregular ruminate" (Figure 11). In much smaller numbers rounded forms were present, including about 11%--"ellipsoid wrinkled", 6%--"spherical granular form", and cylinders ("smooth" and "smooth curved") about 6%. Singular encounter "parallelepiped elongated grungy", "grungy thinned with rounded edges", and "multicellular polyhedra". Morphometric parameters of phytoliths in Amur cork are shown in Table 7.
Viburnum sargentii (Caprifoliaceae)
It grows in Primorye, Amur, Sakhalin, and the South Kuril Islands. It is very close to the plant of used in medicine cranberry tree that on the Far East is not found. Assumed that Sargent cranberrybush viburnum replaces here the cranberry tree and can be used on an equal footing with it. For therapeutic purposes, the bark and fruit of Viburnum is used. One of the important effects of drugs is the bark of Viburnum--increasing of the blood clotting. There is experimental evidence that the blood clotting increases by leaves, flowers, and fruits of this plant. Under experimental conditions, an antimicrobial effect of fruit of Viburnum was revealed .
We investigated the fruit and leaves of viburnum (Figure 12). And in the fruit and leaves prevailed phytoliths of "irregular smooth" and "irregular ruminate"--40%, "smooth curved cylinders" --12%, "grungy thinned parallelepiped with rounded edges"--9%, "the subordinate epidermal the hair"--7%, "ellipsoid wrinkled"--5%, "smooth cylinder"--4%. A small number of "smooth cylinder" (6%), "smooth ellipsoid" (5%), and "a box with attenuated smooth square edges" (4%) was also met. Morphometric parameters of phytoliths of Sargent viburnum are shown in Table 8.
Ribes mandshuricum (Grossulariaceae)
Widespread in China, Korea, and the south of the Far East. Found in separate instances and small thickets. Ribes mandshuricum (Manchu currant) contains vitamins, potassium, magnesium, and manganese. It is used as a general tonic.
Typical composition of phytoliths of currant is slightly different from other samples studied (Figures 13 and 14). The predominant type of phytoliths is "irregular ruminate" (57%), significantly few encountered phytoliths as "thinning rough parallelepiped with rounded edges" (10%) and "smooth ellipsoids" (9%). Phytoliths "irregular smooth" and "cylinders smooth curved" up to 4% of total number of phytoliths. Single or in small amounts met the following types of phytoliths: "adventitious epidermal hair cells", "plate", "short truncated cone-shaped phytoliths", and "smooth cylinders". Morphometric parameters of phytoliths of Manchu currant are given in Table 9.
Aronia melanocarpa (Rosaceae)
Homeland of black chokeberry is the Eastern part of North America. In 1935, it was brought to Altai, and was then extended to other regions of Russia. This plant is applied in therapeutic and prophylactic purposes within pathological conditions involving increased permeability and fragility of capillaries, diabetes, bleeding diathesis, capillary toxicosis, radiation sickness [13, 14].
Samples of black chokeberry are quantitatively poor with phytoliths compared to samples of other studied plants. Morphotypes of phytoliths are also limited compared with other plants, the predominant form that has not been identified. Most often the phytoliths in the form of the "ellipsoid wrinkled" (11%), "smooth cylinder" (9%), "short elongated cells" (6%) were met. Other phytoliths met in approximately equal amounts are in the forms of "thinned parallelepiped with rounded smooth edges" as well as "square-edged", "epidermal adnexal hair cells", "cells truncated short", "spherical granule cells", "polyhedra multicellular", and uncertain morphotypes (Figures 15 and 16). Morphometric parameters of phytoliths of black chokeberry are shown in Table 10.
Panax ginseng (Araliaceae)
It is herbaceous perennial medicinal plant that grows in Asia and North America. It is mainly used as an adaptogen and as a tonic drug, but in recent years was found new uses [15-17]. In Korea and China, ginseng root is also used in cooking.
The most common phytoliths in leaves refers to the types (Figures 17-19) of irregular smooth (35.34%) and irregular ruminate (18%), parallelepiped (16%), smooth oval (13.3%), and lanceolate smooth (10.7%). The least common types are "all smooth" and "long smooth barrel" by 0.33%.
In the roots of ginseng prevailed only one type of phytoliths--irregular ruminate (98.6%) (Figure 19). Isolated finds (1.4%) were of "round smooth" morphotype. Most of "irregular ruminate" phytoliths (85.3%) were very small in size (7.57 [+ or -] 2.09 [micro]m x 9.79 [+ or -] 2.77 [micro]m).
We think this fact deserves serious attention. Given that the medicinal properties and high content of biologically active substances (panaxosides, xatriols, polyacetylenes) has only ginseng root, we can assume that there is a relationship between the biosynthesis of organic components and silicon microcrystals. Morphometric parameters of phytoliths of ginseng are shown in the Table 11.
Images of all phytoliths of investigated plants are shown in Figures 1-19.
This study is the first attempt to describe phytoliths in modern medicinal plants of Primorye. In the process of definition, the species-specific phytoliths have not been identified except stellate type of Amur barberry leaves.
It should be noted that the morphometric analysis of the data gave interesting additional information. First, the size of phytoliths differs by no more than in 10 times. For example, the minimum mean square of phytoliths found in Sargent viburnum (146.39 [micro][m.sup.2]), and the maximum--in black chokeberry (1367.32 [micro][m.sup.2]). The same range of indicators in different plants within the same order is observed for the other morphometric parameters: perimeter, length, and width.
The most important question that the answer must be found--is there a correlation between the type and the number of phytoliths and plant exhibit medicinal properties? We believe that there must be a relationship between metabolism and conjugated silicon biogeochemical cycles, possibly associated with lipid metabolism.
This work was supported by Scientific Foundation of Far Eastern Federal University, President's Grant for young scientists MK-1547.2013.5 and Ministry of Education and Science of the Russian Federation (No. 14.594.21.0006).
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Published: 2nd Nov 2014
Kirill Sergeevich Golokhvast (1) *, Vladimir Victorovich Chaika (1), Sergey Maksimovich Ugay (1), Ivan Vladimirovich Seredkin (2), Igor Eduardovich Pamirsky (3)
(1) Far Eastern Federal University, 8 Sukhanova Street, Vladivostok 690990, Russian Federation.
(2) Pacific Institute of Geography FEB RAS, 7 Radio Street, Vladivostok 690041, Russian Federation.
(3) Blagoveshchensk State Pedagogical University, 50 Lenina Street, Blagoveshchensk 675000, Russian Federation.
Corresponding author: Far Eastern Federal University, 8 Sukhanova Street, Vladivostok 690990, Russian Federation.
Table 1: List of plants of Primorsky region, in which phytoliths were studied. Families Species Parts of the plant Schisandraceae Schisandra chinensis Vine, berry, leaf Saxifragaceae Bergenia pacifica Leaf Berberidaceae Berberis amurensis Leaf Caprifoliaceae Viburnum sargentii Leaf, berry Grossulariaceae Ribes mandshuricum Leaf Rutaceae Phellodendron amurense Fruit Rosaceae Aronia melanocarpa Fruit Rosa acicularis Fruit Araliaceae Eleutherococcus senticosus Leaf Panax ginseng Leaf, root Table 2: Morphometry of phytoliths of Berberis amurensis. Characteristics Quantity of Area Perimeter morphotypes ([[micro] ([micro]m) m.sup.2]) Average 8 648.59 129.04 Minimum 67.13 35.22 Maximum 9035.07 1307.23 Characteristics Width Length ([micro]m) ([micro]m) Average 27.34 45.68 Minimum 9.43 11.72 Maximum 257.89 459.61 Table 3. Morphometry of phytoliths of Schisandra chinensis. Characteristics Quantity of Area Perimeter morphotypes ([[micro] ([micro]m) m.sup.2]) Average 12 473.23 88.41 Minimum 184.04 43.87 Maximum 5021.88 622.4 Characteristics Width Length ([micro]m) ([micro]m) Average 20.79 31.25 Minimum 13.89 17.61 Maximum 114.34 226.62 Table 4: Morphometry of phytoliths of Rosa acicularis. Characteristics Quantity of Area Perimeter morphotypes ([[micro] ([micro]m) m.sup.2]) Average 9 1150.98 165.74 Minimum 308.68 66.99 Maximum 7653.45 922.83 Characteristics Width Length ([micro]m) ([micro]m) Average 60.37 31.93 Minimum 22.89 17.91 Maximum 236.16 136.79 Table 5: Morphometry of phytoliths of Bergenia pacifica. Characteristics Quantity of Area Perimeter morphotypes ([[micro] ([micro]m) m.sup.2]) Average 8 472.99 86.44 Minimum 108.55 43.50 Maximum 6041.47 791.65 Characteristics Width Length ([micro]m) ([micro]m) Average 21.83 30.05 Minimum 12.45 13.93 Maximum 173.73 279.06 Table 6: Morphometry of phytoliths of Eleutherococcus senticosus. Characteristics Quantity of Area Perimeter Width Length morphotypes ([[micro] (Mm) (Mm) (Mm) m.sup.2]) Average 5 362.44 81.16 20.63 28.02 Minimum 156.63 55.82 14.37 21.24 Maximum 1673.64 231.72 59.13 80.98 Table 7: Morphometry of phytoliths of Phellodendron amurense. Characteristics Quantity of Area Perimeter morphotypes ([[micro] ([micro]m) m.sup.2]) Average 8 973.33 132.3 Minimum 234.11 36.72 Maximum 7006.22 836.41 Characteristics Width Length ([micro]m) ([micro]m) Average 49.12 29.78 Minimum 22.97 16.33 Maximum 343.47 145.59 Table 8: Morphometry of phytoliths of Viburnum sargentii. Characteristics Quantity of Area Perimeter morphotypes ([[micro] ([micro]m) m.sup.2]) Average 10 146.39 47.78 Minimum 63.72 30.94 Maximum 239.41 64.55 Characteristics Width Length ([micro]m) ([micro]m) Average 12.69 16.40 Minimum 8.70 10.00 Maximum 16.70 22.26 Table 9: Morphometry of phytoliths of Ribes mandshuricum. Characteristics Quantity of Area Perimeter morphotypes ([[micro] ([micro]m) m.sup.2]) Average 9 371.8 78.39 Minimum 105.79 43.23 Maximum 5165.8 744.82 Characteristics Width Length ([micro]m) ([micro]m) Average 19.84 28.14 Minimum 11.31 15.11 Maximum 176.24 277.86 Table 10: Morphometry of phytoliths of Aronia melanocarpa. Characteristics Quantity of Area Perimeter morphotypes ([[micro] ([micro]m) m.sup.2]) Average 9 1367.32 140.38 Minimum 175.63 47.82 Maximum 102253.7 818.12 Characteristics Width Length ([micro]m) ([micro]m) Average 30.32 52.1 Minimum 14.54 15.21 Maximum 251.58 922.11 Table 11: Morphometry of phytoliths of Panax ginseng. Characteristics Quantity of Area morphotypes ([[micro]m.sup.2]) Average 7 434.08 [+ or -] 255.17 Minimum 162.65 [+ or -] 82.2 Maximum 4857.22 [+ or -] 1710.35 Characteristics Perimeter Width ([micro]m) ([micro]m) Average 90.89 [+ or -] 48.69 15.72 [+ or -] 6.99 Minimum 50.73 [+ or -] 18.02 13.02 [+ or -] 4.1 Maximum 764.09 [+ or -] 260.93 22.42 [+ or -] 6.81 Characteristics Length ([micro]m) Average 27.67 [+ or -] 8.92 Minimum 15.88 [+ or -] 4.63 Maximum 346.45 [+ or -] 113.12
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|Title Annotation:||Research Article|
|Author:||Golokhvast, Kirill Sergeevich; Chaika, Vladimir Victorovich; Ugay, Sergey Maksimovich; Seredkin, Iva|
|Publication:||Biology and Medicine|
|Date:||Jul 1, 2014|
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