Microscopic Analysis of the Aerial Parts of Potentilla bifurca L. Growing in Central Kazakhstan
G.T. Murzaliyeva1, A.M. Essedova2*, B.B. Zhakbayeva3, O. Zhanat3, M.Yu. Ishmuratova4
1Associate Professor, Candidate of Pharmaceutical Sciences, School of Pharmacy,
NJSC Karaganda Medical University, 100008, st. Gogol 40, Karaganda, Republic of Kazakhstan.
2Masters Degree, School of Pharmacy, NJSC Karaganda Medical University,
100008, St. Gogol 40, Karaganda, Republic of Kazakhstan.
3School of Pharmacy, NJSC Karaganda Medical University,
100008, st. Gogol 40, Karaganda, Republic of Kazakhstan.
4Professor, Candidate of Biological Sciences, NJSC Karaganda University Named after academician E.A. Buketov, 100026, st. Universitetskaya 28, Karaganda, Republic of Kazakhstan.
*Corresponding Author E-mail: esedova.aizhan@mail.ru
ABSTRACT:
This study presents the results of a detailed microscopic analysis of the aerial parts of Potentilla bifurca L., collected in June 2022 and 2024 from the Karkaraly Mountains (Karkaraly district, Karaganda region). The materials studied included leaves, inflorescences, and stems, which underwent thorough investigation using an Altami scanning microscope equipped with a digital camera (3.1 MP) at magnifications of 16ื4 and 16ื10. The analysis provided comprehensive insights into the anatomical structure of the plant, allowing for the identification of several key distinguishing features. During the microscopic examination, we identified the shape of epidermal cells on the leaves and flower petals, as well as the presence of both unicellular and multicellular trichomes on the stems, leaves, and peduncles. The form and arrangement of calcium oxalate druses in the leaf cells were also observed, providing additional structural details. Furthermore, an anomocytic stomatal complex was documented, which is a typical feature of this species. Special attention was given to the stem structure, where the complete absence of cambium was noted, marking it as an important anatomical diagnostic feature. Histochemical analysis of the aerial parts of the plant confirmed the presence of phenolic compounds and flavonoid groups in the aerial parts of the plant. The results of this study suggest that the identified microscopic features of Potentilla bifurca L. are not only consistent but also reliable for the accurate identification of this plant material, supporting its potential use in further pharmacological and botanical research.
KEYWORDS: Potentilla bifurca L., Microscopy, Aerial parts, Diagnostic features, Trichomes, Stomata.
INTRODUCTION:
The genus Potentilla L. is one of the largest genera in the Rosaceae family, comprising approximately 500 species, primarily distributed across the Northern Hemisphere, especially in temperate and subtropical regions. Potentilla species are of significant interest for medicine and pharmaceutical production due to their wide range of biologically active compounds. Several representatives of the genus have long been used in traditional medicine. However, the identification and chemical composition of the raw plant materials remain insufficiently studied, which limits the clear interpretation of their pharmacological effects and the establishment of their role as pharmacopoeial plants1,2.
Currently, the only representative of the genus described in the State Pharmacopoeia of the Russian Federation and other CIS countries is Potentilla erecta L.
Potentilla bifurca L. is a low-growing perennial shrub, reaches a height of 225cm, with woody underground stems and predominantly ascending or sprawling aerial stems. Previous morphological studies of this species have demonstrated its similarity to other members of the genus, characterized by a high degree of polymorphism depending on environmental conditions3,4.
In 2007, L.I. Sdobnina and E.Yu. Frolova described the anatomical and morphological features of plant materials collected near Penza, noting diagnostic characteristics in the stems, petioles, and epidermis5. Our study extends these investigations to include the upper and lower epidermis of leaves, transverse sections of leaves, stems, peduncles, sepals, and flower petals.
MATERIALS AND METHODS:
Collection and Identification of Plant Material:
The plant material used in this study comprised the aerial parts (leaves, inflorescences, and stems) of Potentilla bifurca L. collected in June 2022 and 2024 from the Karkaraly Mountains (Karkaraly district, Karaganda region).
Microscopic Analysis:
Samples of raw plant material (leaves, stems, and flowers) were fixed in a mixture of glycerin:96% ethanol:distilled water in a 1:1:1 ratio (Strauss-Fleming solution)6,7. Cross-sections of leaves, petioles, and stems were prepared manually. Microscopic slides of stems were made from the middle part of generative shoots, for leaves from the central part of the terminal leaf fragment, and for sepals from the middle region8,9. Surface preparations of the upper and lower epidermis of leaves and flower petals were also made. Sections were clarified using glycerin and photographed with the Altami scanning microscope at 16ื4 and 16ื10 magnifications10,11. Measurements and image processing were performed using Altami Studio and PhotoScape software12,13,14.
Histochemical Analysis:
Histochemical analysis was conducted on slides prepared from aerial parts of Potentilla bifurca L.15,16,17. Freshly prepared reagents were applied to detect specific chemical compounds, and the resulting images were captured using the Altami scanning microscope18,19,20.
RESULTS AND DISCUSSION:
Microscopic Analysis:
Anatomical Structure of the Leaf
The upper and lower epidermis of Potentilla bifurca L. leaves consists of oval cells with slightly wavy walls (Figure 1), covered by a thin cuticle layer. Epidermis cells over the veins are elongated with nearly straight walls (Figure 2). Few simple trichomes are scattered across the surface, with their bases forming rosettes of adjoining epidermis cells. The stomata are small, sparse, and of the anomocytic type, with a stomatal index of 13.3. Calcium oxalate druses are rounded and darkly stained, are well visible.
Figure 1. Upper (left) and lower (right) epidermis of Potentilla bifurca L. leaves. Magnification: 16ื10. 1 - calcium oxalate druses, 2 - trichome base, 3 - epidermal cells, 4 - trichome.
Figure 2. Surface preparation of Potentilla bifurca L. leaf. Epidermal region above the midvein. Magnification: 16ื10.
1 - Epidermal cells above the leaf vein, 2 - primary epidermal cells.
The leaf in cross-section is flat and dorsiventral (Figure 3). The cells of the upper and lower epidermis are oval in shape, with the cells of the lower epidermis being larger than those of the upper. A cuticle layer is observed on both sides. Trichomes are simple, thin, and can be unicellular or multicellular, predominantly located on the lower side of the leaf. The palisade mesophyll is two-layered, while the spongy mesophyll is multilayered. The vascular bundles are small, collateral, and of a closed type. Small calcium oxalate druses are visible in the mesophyll.
Figure 3. Cross-sections of Potentilla bifurca L. leaf (left) and sepal (right). Magnifications: 16ื10 and 16ื4, respectively. 1 - trichome, 2 - upper epidermis, 3 - palisade mesophyll, 4 - spongy mesophyll, 5 - lower epidermis.
Sepal:
The sepal of Potentilla bifurca L. is flat and isolateral (Figure 3). The cells of the upper and lower epidermis are rounded-rectangular, with sparse simple trichomes. The surface is covered with a cuticle layer. Beneath the epidermis on both sides lies a single-layered palisade mesophyll, with spongy tissues being almost absent. The vascular bundles are small, rounded, collateral, and of a closed type.
Anatomical Structure of the Peduncle:
The peduncle of Potentilla bifurca L. is circular in cross-section (Figure 4). Its periphery is surrounded by a single-layered epidermis with sparse, simple, multicellular trichomes measuring 1020ตm in length. The epidermal cells are rounded-rectangular with thickened outer walls, and the surface is covered by a cuticle layer.
Beneath the epidermis lies a multilayered chlorenchyma, 68 ตm thick. Below the chlorenchyma is a layer of cortical parenchyma, consisting of large, loosely arranged cells. The vascular zone is separated from the cortex by a single-layered endodermis. The vascular zone is of a non-bundled type, represented by a ring of phloem and a ring of xylem vessels. Above the phloem are strands of sclerenchyma. The central part of the peduncle is filled with cells of medullary parenchyma.
Figure 4. Cross-sections of the peduncle (left) and stem (right) of Potentilla bifurca L. Magnifications: 16ื4 and 16ื10, respectively. 1 - trichome, 2 - epidermis, 3 - chlorenchyma, 4 - xylem, 5 - phloem, 6 - endodermis, 7 - medullary parenchyma, 8 - sclerenchyma.
Stem:
The stem of Potentilla bifurca L. in cross-section is round with small ribs (Figure 4). Epidermal cells are bead-shaped with thickened outer walls. Sparse simple trichomes are observed on the surface. Beneath the epidermis lies chlorenchyma, consisting of 23 layers of cells, with localized areas of angular collenchyma. Below the chlorenchyma is a multilayered parenchyma. The vascular system is of a ring type, consisting of a ring of sclerenchyma, a thin layer of phloem, and chains of xylem. Cambium is absent. The central part of the stem is filled with loose medullary parenchyma cells.
Flower Petal
The petal consists of small oval epidermal cells tightly adjacent to each other (Figure 5). Darkly stained veins and occasional pollen grains are well-visible on the surface. Sparse simple trichomes are scattered across the surface, with their bases forming "rosettes" of adjoining epidermal cells. Stomata are small, sparse, and of the anomocytic type (Figure 5).
Figure 5. Fragment of the corolla and flower preparation of Potentilla bifurca L. from the surface. Magnification: 10ื16. 1 - primary epidermis cells, 2 - pollen grains, 3 - trichome base, 4 - trichome, 5 - vein.
Microscopic examination of the aerial parts of Potentilla bifurca L. revealed the following structural features:
The shape of epidermal cells in the leaves and flower petals.
The presence of simple unicellular and multicellular trichomes on the stem, leaf, and peduncle preparations.
The form and distribution of calcium oxalate druses in the leaf structure.
Compared to Potentilla bifurca L. from Penza, the material studied in this research exhibited an anomocytic stomatal complex, absence of cambium in the stem, and lack of a described petiole.
Histochemical Analysis:
The preparations of the aerial part of Potentilla bifurca L. exhibit characteristic chemical reactions during histochemical analysis with the use of various reagents, resulting in specific color changes (Table 1).
Table 1. Histochemical Analysis of Aerial Parts of Potentilla bifurca L.
|
S. No. |
Components |
Reagents |
Staining Color |
Stem |
Peduncle |
Leaf |
Sepal |
Petal |
|
1 |
Essential oils |
Methylene blue |
Blue |
- |
- |
- |
- |
- |
|
2 |
Sesquiterpene lactones |
Vanillin + conc. H2SO4 |
Yellow |
- |
- |
- |
- |
- |
|
3 |
Flavonoids |
1% alcohol solution of FeCl3 |
Black-blue-green |
+ |
+ |
+ |
+ |
+ |
|
4 |
Phenolic compounds |
10% alcohol solution of K2Cr2O7 |
Brown, yellow |
+ |
+ |
+ |
+ |
+ |
|
5 |
Phenolic compounds |
3% alcohol solution of FeCl3 |
Yellow-brown |
+ |
+ |
+ |
+ |
+ |
|
6 |
Polysaccharides |
10% solution of thymol + conc. H2SO4 |
Orange-red |
- |
- |
- |
- |
- |
|
7 |
Starch |
Lugol's reagent |
Blue |
- |
- |
- |
- |
- |
|
8 |
Alkaloids |
Dragendorff's reagent |
Black |
- |
- |
- |
- |
- |
CONCLUSION:
This study encompasses the microscopic analysis of the aerial organs of Potentilla bifurca L. Distinctive microscopic features were identified, including the shape of epidermal cells in leaves and flower petals, the presence of trichomes on stems, leaves, and peduncles, and the form and arrangement of calcium oxalate druses in leaf structure. Histochemical analysis confirmed the presence of phenolic compounds and flavonoid groups in the aerial parts. These diagnostic features establish a reliable basis for the identification of Potentilla bifurca L. as a raw plant material.
CONFLICT OF INTEREST:
No conflict of interest declared.
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Received on 24.12.2024 Revised on 21.04.2025 Accepted on 25.06.2025 Published on 01.10.2025 Available online from October 04, 2025 Research J. Pharmacy and Technology. 2025;18(10):4977-4980. DOI: 10.52711/0974-360X.2025.00719 ฉ RJPT All right reserved
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