Journal of Medicinal Plants Biotechnology

Abstract The classical methods of plant genetic transformation have bottlenecks to achieve plants with interesting genotypes, while achieving remarkable success. In the process of gene transfer to plants, biomolecules must be transferred to the plant cell nucleus by passing through the tough and multi-layered plant cell wall. Nanotechnology has been widely used in medicine and life sciences, and various nanomaterials such as single walled carbon nanotubes (CNTs) have been used as non-viral carriers for the transfer of biomolecules for targeted delivery. In this study, nanocarriers were prepared after functionalization of SWCNTs and changing the surface of SWCNTs. Then pDNA containing mgfp5-ER reporter gene was loaded on the functionalized SWCNTs. In the next step, gene transfer to plant cells was carried out by the prepared SWCNTs@pDNA nanocarrier. The success of gene transfer was evaluated by the fluorescent signal of mgfp5-ER reporter gene. The results shown the preparation of a nanocarrier with a high capacity to loading pDNA on its surface and the ability of the prepared nanocarrier to pass through the cell wall and successfully transfer pDNA into the plant cell. So, this method can be considered as a relatively simple method, independent of the plant species and without the need for special laboratory equipment for gene transfer to walled plant cells. Therefore, nanobiotechnology and nanomaterials can be a window of hope for improving and increasing the efficiency of conventional methods of gene transfer to plant cells.

Abstract Plants synthesize a huge variety of secondary metabolites with complex chemical composition that not only have medicinal value but are also important to deal with biotic and abiotic stress conditions. The aim of current study was to investigate the impact of heat shock treatment on the production of secondary metabolites in callus obtained from of Echinacea purpurea plant. For this purpose, seeds were grown on MS medium. Then root explants were prepared from 45 days seedlings and transferred to MS medium containing 0/5 and 1 mg/L 2,4-D and Kin. Heat shock treatment at 35 °C for different time durations (10, 20, 30, 40, 50 and 60 min) was daily applied on 2 months old callus. After a week, the samples were collected and different biochemical analysis was performed. The results showed that the highest amount of total phenol, total flavonoid, soluble suger, MDA, chicoric acid, chlorogenic acid and caffeic acid was observed in samples that were treated for 60 min at 35 °C. On the other hand, the present data revealed that heat shock treatment caused a significant increase in catalase and peroxidase activity, but there was not any significant difference between different treatments. As a conclusion, heat shock treatment can be used as a new method to enhance secondary metabolites production in Echinacea in tissue culture contion.

Abstract Drogonhead from the Lamiaceae family is an important plant with many uses . This research was performed in pots and done in a randomized complete block design (RCBD) in a split-split plot layout with three replications. Irrigation regimes (D1-D3: Field Capacity, 50% and 75% evaporation from basin class A) in main plots, superabsorbent nano-polymer application (S1-S2: use and not use) in sub plots and treatments of fertilizers in sub-sub plots were done. Over two consecutive years, the obtained results showed that the use of treatments increased the characters and made the significant effectiveness. In two years, the highest shoot dry matter (54.43 g.plant-1), zinc of shoot (68.12 µg/g dry weight), iron of shoot (75.68 µg/g dry weight), phenol (1.22 mg.Kg Fw-1), chlorophyll content (2.55 mg.g-1 DW) and essential oil content (1.12 %) made by grow-more fertilizers specially grow-more 1 (20-20-20) in combination with Fe/Zn Libfer with irrigation regime of 25% evaporation. The lowest values for morpho-physiological and phytochemical characters such as shoot dry matter (19.82 g.plant-1), zinc of shoot (25.43 µg/g dry weight), iron of shoot (38.43 µg/g dry weight), phenol (0.33 mg.KgFw-1), chlorophyll content (1.88 mg.g-1 DW) and essential oil content (0.28 %) obtained by plants treated with irrigation regime of 75% evaporation from basin class A (D3) and no application of fertilizers. Generally the application of grow-more fertilizer 1 (20-20-20) with consumption of zinc and iron fertilizers can be a good strategy to improve morpho-physiological characters and essential oil quantity and quality of D.moldavica in same climates.

Abstract Assessing the genetic diversity in the population is the prerequisite to start and develop plant breeding projects. The genus Hedera L. occupies forest understories and riparian vegetation in temperate latitudes throughout Europe, N Africa and Asia. Therefore, we collected and analyzed 35 Hedera genotype, from five provinces of Iran regions. Our aims were 1) to assess genetic diversity among some of Irainian Hedera cultivars 2) is there a correlation between species genetic and geographical distance? 3) Genetic structure of populations and taxa. We showed significant differences in quantitative morphological characters in plant species. Hedera colchica depicted unbiased expected heterozygosity (UHe) in the range of 0.19. Shannon information was high (0.39) in Hedera helix cultivars. Hedera colchica cultivars showed the lowest value, 0.23. Gene flow (Nm) was relatively low (0.29) in Hedera cultivars. The Mantel test showed correlation (r = 0.86, p=0.0001) between genetic and geographical distances. We reported high genetic diversity, which clearly shows the among some of Irainian Hedera cultivars can adapt to changing environments since high genetic diversity is linked to species adaptability. Present results highlighted the utility of RAPD markers and morphometry methods to investigate genetic diversity in Hedera cultivars.

Abstract The Ferula gummosa plant is a perennial and valuable species from the Apiaceae family, native to Iran. The significance of this plant lies in its resin, known as galbanum. Dormancy of the seeds is one of the main challenges in its growth, taking at least 9 months under natural conditions. The aim of this experiment was to shorten the dormancy period of this plant using artificial seed technology. In this experiment, somatic embryos of F .gummosa at the heart-shaped and torpedo stages were obtained from leaf explants in an MS1/2 medium containing 1 mg/L of BAP hormone and 1 mg/L of NAA hormone. Additionally, the embryonic part of the seed was isolated and encapsulated using a 2% sodium alginate solution, resulting in the production of artificial seeds. The results showed that the encapsulated somatic embryos at the torpedo stage showed the greatest seedling length after 15 and 30 days, and the germination of these seeds was faster compared to other seeds. The seeds obtained from the embryonic part also showed acceptable results. The findings of this experiment indicate that considering the time required for producing artificial seeds of F. gummosa, this method is quicker and more suitable for the propagation of this plant.

Abstract

Abstract Borage, generally known as Iranian Borage flower (scientific name Echium amoenum Fisch. & C.A. Mey.), is among the important, well-known, and widely used medicinal plants in human medicinal history and holds a special place in various cultures and folklores, particularly in Iran. This plant appears to rank after thyme in the list of the most consumed plants in Iran. Across the world, the flowers and leaves of E. amoenum are utilized for medicinal purposes, including the treatment of stress, cardiovascular diseases, cough, and lung disorders, as well as for their tonic and sedative properties. This plant is native to limited regions of northern and northwestern Iran and the Caucasus and it also grows in many parts of Europe, Western Asia, and North America, where it is often found growing wild along riverbanks. Iranian borage is considered one of the most valuable and popular medicinal plants in Iran. The Boraginaceae family comprises more than 130 genera and 2300 species. The Echium genus includes 67 species. According to global research, Iranian cowpea contains natural bioactive compounds with health-promoting effects, including antioxidant, antibacterial, antiviral, antidiabetic, anti-inflammatory, soothing, and immunoregulatory properties. In traditional medicine, the petals of this plant are used as a diuretic, pain reliever, diaphoretic, and blood pressure reducer. The principal constituents of this plant are polyphenols, rosmarinic acid, and flavonoids. This plant has seeds rich in alpha-linolenic and gammalinolenic essential fatty acids, which are among the fatty acids required for the formation of prostaglandins involved in the synthesis of the nerve myelin sheath. For this reason, it is used in the preparation of medicinal supplements to prevent neurological diseases such as MS. Genetic diversity, as well as the use of biotechnological methods such as cell culture and callus culture in this plant, is among the approaches for producing secondary metabolites that has been studied by numerous researchers and is discussed in this article. Most of the conducted research has been based on micro-proliferation. The nutritional medium used was Murashige and Skoog, and most of the hormones applied were 42-dichloroethoxyacetic acid, naphthalene acetic acid, and 6-benzylaminourea, or a combination of them.

Abstract Helicobacter pylori, a Gram-negative bacterium, is a major causative agent of peptic ulcers and gastrointestinal cancers. To neutralize gastric acid and ensure survival in the harsh stomach environment, H. pylori relies on urease enzyme activity, which converts urea into ammonia. Hence, urease inhibitors play a critical role in combating H. pylori and managing associated diseases, including gastritis and other gastrointestinal disorders. This study evaluated the urease-inhibitory and antioxidant activities of Cydonia oblonga Mill. (quince) fruit and seed gel extract, aiming to identify natural sources of effective urease inhibitors. The urease inhibition potential was assessed using the Berthelot method (targeting jack bean urease), while antioxidant activity was measured via the 2, 2-Diphenyl-1-Picrylhydrazyl (DPPH) free radical scavenging assay. Both extracts were prepared with 80% ethanol, and inhibitory effects were quantified through enzyme inhibition percentages and IC₅₀ values for urease, alongside DPPH radical scavenging percentages. The results revealed that the seed gel extract exhibited stronger urease inhibition (IC₅₀ = 136.6 ± 7.5 μg/mL) compared to the fruit extract (IC₅₀ = 435.3 ± 10.8 μg/mL). Antioxidant activity was higher in the fruit extract (85.23%) than in the seed gel (79.66%). These findings highlight Cydonia oblonga through its suitable antioxidant, and urease-inhibitory activity for developing safer therapeutic strategies against H. pylori infection and preventing relevant complications.

Abstract Salinity stress, as one of the major challenges in agriculture, disrupts water and nutrient uptake by accumulating sodium and chloride ions in the soil, leading to reduced plant growth and yield. This stress damages plant cells through oxidative stress and disrupts metabolic processes. In this study, the effects of different salinity levels (0, 2.5, and 5 dS/m) and salicylic acid foliar application (0, 0.5, and 1 mM) on purslane (Portulaca oleracea) were investigated using a completely randomized factorial design with three replications. The results showed that salinity reduced plant growth and negatively affected chlorophyll a and b content. However, the application of salicylic acid (1 mM) mitigated these adverse effects by improving potassium uptake. Although salinity increased sodium and chloride accumulation, salicylic acid improved plant tolerance by reducing oxidative stress and maintaining photosynthetic efficiency. In conclusion, this study demonstrates that salicylic acid can serve as an effective strategy to enhance purslane's resistance under saline conditions.

Abstract The medicinal plant garlic (Allium sativum L.) is known for its synthesis of allicin, a defense molecule that exhibits various biological activities. Traditional medicine uses this medicinal plant to alleviate numerous diseases due to the wide range of effects of garlic. Allicin is produced from alliin during the cutting of garlic by the activity of the enzyme allinase and is effective against a wide range of microorganisms. Allicin is hydrophobic in nature, can efficiently cross cell membranes, and behaves as an active sulfur species inside cells. It is a physiologically active molecule with the ability to oxidize the thiol groups of glutathione and between cysteine residues in proteins. Allicin is physiologically active in microbial, plant, and mammalian cells. Allicin can inhibit the proliferation of bacteria and fungi in a dose-dependent manner or kill cells completely. In addition, in mammalian cell lines, including cancer cells, allicin causes cell death and inhibits cell proliferation. In plants, allicin prevents seed germination and reduces root growth. The study showed that allicin has a wide range of pharmacological activities. Allicin has antimicrobial, antioxidant, anticancer properties, the ability to reduce cardiovascular diseases, improve the immune system and regulate blood sugar. The antimicrobial and antioxidant functions of allicin are due to the reaction of allicin with the thiol group of various enzymes, respectively; to inhibit the metabolism of cysteine protease and to trap free radicals. Allicin prevents the occurrence and progression of cancer by blockin

Abstract Catharanthus roseus is an important medicinal plant containing alkaloids such as vinblastine and vincristine, which have strong cell division inhibitory activity and can serve as a colchicine alternative in polyploidization programs. This study aimed to evaluate the effects of aqueous and ethanolic leaf extracts of C. roseus on cell division in meristematic cells of onion (Allium cepa) roots. Extracts at four concentrations (0, 20, 50, and 100%) were applied to onion bulbs. The experiment was conducted as a completely randomized factorial design with three replications. Traits assessed included root length and number, rooting time, mitotic and phase indices, chromosome counts, and cytological abnormalities such as multinucleation and chromatid bridges. Results showed that increasing extract concentration reduced root length, and rooting was delayed in the ethanolic extract, with the greatest delay observed at 100% concentration. The highest prophase index was recorded at 20% extract for both types, while the highest telophase index occurred at 100% ethanolic extract. Mitotic index peaked at 20% and 50% ethanolic extracts. Chromosomal abnormalities, including stickiness, breakage, and duplication, were mainly observed in 50% ethanolic extract, whereas chromatid bridges appeared only in 50% aqueous extract. These findings indicate that C. roseus alkaloids can influence mitosis and may be applied in breeding programs and polyploidization of medicinal plants.

Abstract Abstract
Plant tissue culture is recognized as a key tool in various research fields, especially in the area of medicinal plants. These techniques are employed for mass propagation, conservation, and production of secondary metabolites in medicinal plants. There are various methods for in-vitro culture, including micropropagation, axillary bud culture, organ culture, root and callus culture, organogenesis, somatic embryogenesis, and cell suspension culture. Since cell suspension culture and callus are typically preferred for producing plant chemicals, following root and shoot cultures as well as somatic embryogenesis, these methods play a significant role in optimizing the production of secondary metabolites. However, one of the major challenges in plant tissue culture is the potential occurrence of somaclonal variation, which can result from genetic mutations or changes in epigenetic markers. These variations particularly arise in highly differentiated explants and during the callus stage. Additionally, the occurrence of somaclonal variation may pose a barrier to successful in-vitro propagation and preservation of germplasm. This issue is especially critical in cases where maintaining the genetic and biochemical characteristics of medicinal plants is important. In the present study, the potential somaclonal variations resulting from the tissue culture of medicinal plants and their implications for the production of secondary metabolites are examined and discussed. This research can contribute to a better understanding of the challenges and opportunities associated with the use of tissue culture for the production of medicinal plants and valuable metabolites.