Source Of Traditional Medicine Research Articles

Greek flora as a source for the detection of natural compounds potentially effective in preventing post-menopausal osteoporosis

Osteoporosis is the most common metabolic bone disorder and remains an increasingly significant problem affecting post-menopausal women. It results from an imbalance between the processes of bone formation and resorption leading to decreased bone mass and increased risk of fracture. Treatments against osteoporosis include hormone replacement therapy (HRT) with estrogens, that unfortunately is combined to an increased hormone-dependant cancer risk, and selective estrogen receptor modulators (SERMs). Nowadays, numerous women use plant derived supplements to prevent postmenopausal diseases including osteoporosis. Despite Greece constituting one of the richest floral diversity regions, there are scarcely any reports connecting Greek flora herbs with prevention of osteoporosis. Therefore, the aim of this work is to identify and characterize plant extracts deriving from Greek flora, and isolated compounds that display significant SERM activity. Considering our research experience on natural products with estrogenic activities and traditional medicine sources, 44 plant species were selected and 91 extracts were prepared using conventional (maceration) and modern-green techniques (ASE, MAE and SFE). Preliminary phytochemical and biological screening revealed Rhamnus sp, Lupinus album, Psoralea bituminosa, Ceratonia siliqua and Cytisus villosus between the most capable extracts of inducing MC3T3-E1 differentiation to osteoblasts. Consequently, we proceeded to the isolation of the major compounds, which were structurally elucidated using NMR spectroscopy, and in vitro evaluated using a cell-based screening, concerning the differentiation of MC3T3-E1 cells to osteoblasts and their estrogenic-antiestrogenic properties. The obtained results, revealed derivatives of genistein, kaempferol and its glycosides, as the most promising agents for the potential prevention and treating of post-menopausal osteoporosis.

Herbal Remedies for Functional Dyspepsia and Traditional Iranian Medicine Perspective.

Context:Functional dyspepsia (FD) is a functional gastro-intestinal disorder with high prevalence. Among various treatment options, treatment by complementary and alternative medicines especially herbal remedies also practiced. Traditional Iranian medicine (TIM), a valuable resource of valid applied studies of ancient Iranian scholars, recommends numerous medicinal plants to treat dyspepsia symptoms. In this study, through investigation of TIM references, we aimed to identify medicinal plants for treatment of digestion insufficiency.Evidence Acquisition:In this qualitative study, dyspepsia symptoms including fullness, early satiety, bloating, nausea, and belching were checked under reliable sources of traditional medicine. Then medicinal plants recommended for the treatment of the symptoms were extracted from the books. Likewise, for investigating the pharmacological properties of medicinal plants used for the relieving of dyspepsia symptoms, electronic databases such as PubMed, Scopus, Google Scholar and some Iranian databases like SID and IranMedex were employed.Results:The study yielded 105 plants from 37 families which could treat various dyspepsia symptoms; fifty-seven plants, mainly from Apiaceae, Lamiaceae, Amaryllidaceae and Zingiberaceae had digestive effects. In this research, based on the information in TIM reference texts, we obtained 58 plants effective for bloating, 40 for nausea, 37 for appetite loss and 7 for belching. In human clinical trials conducted on medicinal plants effective for FD symptoms, 7 single plants were used.Conclusions:Finding the medicinal plants effective on digestion insufficiency based on TIM could suggest a better strategy for the relieving of dyspepsia symptoms. Traditional Iranian medicine prescribes medicinal plants based on each patient’s personal characteristics and practices multiple target therapies.

Neuropharmacological Aspects of Crocus sativus L.: A Review of Preclinical Studies and Ongoing Clinical Research.

Crocus sativus L. (Iridaceae) is an important member of the genus Crocus having high medicinal value. Its dried stigmas, known as "saffron" are being widely used form past many centuries as a food additive, coloring agent, flavoring agent and a potential source of traditional medicine. The stigmas along with other botanical parts of Crocus sativus are being extensively used in ethnomedical treatment of varied central nervous system diseases. In line with its ethnomedical importance, several preclinical studies have been carried out to validate its traditional uses, identify active principle(s), understand pharmacological basis of therapeutic action and explore novel medicinal uses. The bioactive components of Crocus sativus have been found to modulate several synaptic processes via direct/indirect interplay with neurotransmitter receptor functions, interaction with neuronal death/survival pathways and alteration in neuronal proteins expression. Many clinical studies proving beneficial effect of Crocus sativus in depressive disorders, Alzheimer's disease and some other neurological abnormalities have also been carried out. Based on the vast literature reports available, an attempt has been made to comprehend the fragmented information on neuropharmacological aspects, chemistry and safety of Crocus sativus. Although the plant has been well explored, but still a large scope of future preclinical and clinical research exist to explore its potential in neurological diseases, that has been discussed in depth in the present review.

Comparative analysis of the antibacterial, antifungal, antiproliferative and cyclic response element (CRE) induced expression of downstream luc gene activities of Monopterus albus and Channa straitus extracts

Background: In Malaysia, Monopterus albus is commonly found in rice fields, muddy ponds and swamp areas. Channa striatus have been widely used as a source of traditional medicines. The extracts of Malaysian local Monopterus albus and Channa straitus have been reported to have different bioactive properties and these properties can be used at molecular level as alternative tool for different disorders. Objectives: The comparative analysis of antibacterial, antifungal, antiproliferative and CRE induced expression of downstream luc gene activities of both the extracts were performed. Results: The bacteriostatic and antibacterial effects of both extracts were revealed beside higher antifungal activity of Channa straitus. The extracts from Monopterus albus showed higher levels of antiproliferative activity as compare to Channa straitus. The results were found supportive towards up regulation of hrluc by Monopterus albus extracts and down regulation by Channa striatus extracts. Novelty: This is the first report on comparing the bioactive properties of Monopterus albus and Channa straitus. Future scope: The results from this study demands a further research on identifying the bioactive molecules involve in these actions at a molecular level.

Knowledge and uses of African pangolins as a source of traditional medicine in Ghana.

Traditional medicine has been practised in Ghana for centuries with the majority of Ghanaians still patronising the services of traditional healers. Throughout Africa a large number of people use pangolins as a source of traditional medicine, however, there is a dearth of information on the use of animals in folk medicine in Ghana, in particular the use of pangolins. The aim of this study was to determine the prevalent use of pangolins and the level of knowledge of pangolin use among traditional healers in Ghana for the treatment of human ailments. Data was gathered from 48 traditional healers using semi-structured interviews on the traditional medicinal use of pangolin body parts in the Kumasi metropolis of Ghana. The cultural importance index, relative frequency of citation, informant agreement ratio and use agreement values were calculated to ascertain the most culturally important pangolin body part as well as the level of knowledge dissemination among traditional healers with regards pangolin body parts. Our study revealed that 13 body parts of pangolins are used to treat various medicinal ailments. Pangolin scales and bones were the most prevalent prescribed body parts and indicated the highest cultural significance among traditional healing practices primarily for the treatment of spiritual protection, rheumatism, financial rituals and convulsions. Despite being classified under Schedule 1 of Ghana’s Wildlife Conservation Act of 1971 (LI 685), that prohibits anyone from hunting or being in possession of a pangolin, our results indicated that the use of pangolins for traditional medicinal purposes is widespread among traditional healers in Ghana. A study on the population status and ecology of the three species of African pangolins occurring in Ghana is urgently required in order to determine the impact this harvest for traditional medical purposes has on their respective populations as current levels appear to be unmonitored and unsustainable.

Traditional Uses, Phytochemistry, and Pharmacology of Olea europaea (Olive).

Aim of the Review. To grasp the fragmented information available on the botany, traditional uses, phytochemistry, pharmacology, and toxicology of Olea europaea to explore its therapeutic potential and future research opportunities. Material and Methods. All the available information on O. europaea was collected via electronic search (using Pubmed, Scirus, Google Scholar, and Web of Science) and a library search. Results. Ethnomedical uses of O. europaea are recorded throughout the world where it has been used to treat various ailments. Phytochemical research had led to the isolation of flavonoids, secoiridoids, iridoids, flavanones, biophenols, triterpenes, benzoic acid derivatives, isochromans, and other classes of secondary metabolites from O. europaea. The plant materials and isolated components have shown a wide spectrum of in vitro and in vivo pharmacological activities like antidiabetic, anticonvulsant, antioxidant, anti-inflammatory, immunomodulatory, analgesic, antimicrobial, antiviral, antihypertensive, anticancer, antihyperglycemic, antinociceptive, gastroprotective, and wound healing activities. Conclusions. O. europaea emerged as a good source of traditional medicine for the treatment of various ailments. The outcomes of phytochemical and pharmacological studies reported in this review will further expand its existing therapeutic potential and provide a convincing support to its future clinical use in modern medicine.

Antimicrobial assay and phyto-chemical analysis of Solanum nigrum complex growing in Kenya

Solanum nigrum complex is a group of plants used as indigenous vegetables and also a source of traditional medicine in Kenya and other parts of the world. The main objective of this study was to determine the antimicrobial property and phytochemical composition of S. nigrum complex. Samples of S. nigrum complex were collected from Kenya. The plants were dried under the shade and then ground to a fine powder. Crude extracts were prepared from the plants using methanol and evaluated for antifungal and antibacterial activities against the fungi Fusarium culmorum, Fusarium avenaceum, Fusarium moniliforme and bacteria Staphylococcus aureus, Salmonella typhi, Pseudomonas aureginosa, Proteus mirabilis, Shigella spp., Pseudomonas syringae, Escherichia coli and Basillus subtilis. Minimum inhibition concentration (MIC) of the samples against active microbes was determined. Phytochemical composition was analysed. T-test was used to analyse the significance of the activity indexes of the extracts against the different groups of the microbes. The samples collected were found to belong to the species S. nigrum, Solanum scabrum and Solanum villosum which all belong to the S. nigrum complex. The samples showed a considerable antibacterial and antifungal activity against the tested microbes. The highest antibacterial activity was 29.00 mm which was shown by S. villosum species against S. aureus. The antibacterial and antifungal activities were dose dependent with low MIC values as such 0.09 g/ml given by S. nigrum Mill species against all eight tested bacteria. The antimicrobial activity was associated with the wide array of phytochemical compounds observed in the samples and they include tannins, saponins, flavonoids, steroids, terpenes, phenolic compounds, alkaloids and cardiac glycosides. It was concluded that the S. nigrum complex is rich in many phytochemical compounds like saponins, flavanoids, steroids, glycosides, terpenoids among others which are responsible for the antimicrobial activities of the plants. It can be used not only in treatment of plant fungal infections but also in the management of bacterial human diseases and further research is recommended. Key words: Solanum nigrum complex, phytochemical, antimicrobial.

노니 뿌리 추출물의 매염제 및 매염 조건별 실크 염색성 고찰

Morinda citrifolia, commercially known as Noni, is a tree that grows widely throughout the Pacific, and is recognized as one of the most significant sources of traditional medicines among Pacific Island societies. All parts of the plant have traditional and/or modern uses, including roots and bark(dyes, medicine), trunks(firewood, tools), and leaves and fruits(food, medicine). The bark and the roots of the tree contain red and yellow pigments respectively, which are both used in the manufacture of dyes. Dyes from Noni are being used traditionally, to color clothing and fabrics. In this study, mordanting effects on the dyeing properties of Noni root extract on silk fabrics were investigated.

Genetic analysis of litchi (Litchi chinensis Sonn.) in southern China by improved random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR)

Litchi (Litchi chinensis Sonn., L. chinensis), a type of tree growing in most areas of southern China, produces an edible fruit that is also a source of traditional medicine. Genetic identification of litchi species or cultivars using molecular markers is very important. In this study, a total of six litchi samples from Fujian, Hainan, Guangdong, Guangxi and Sichuan province, as well as one wild Dimocarpus confinis (D. confinis) sample from Guangxi province were collected for genetic analysis. The cluster dendrograms were constructed for genetic analysis on the basis of DNA amplification results by RAPD and ISSR. The improved RAPD amplified DNA with consistent and clear banding patterns. A total of 176 bands were found, indicating a 72.7 % polymorphism in L. chinensis DNA samples. Significant genetic distances were found among the different species or cultivars, with an index of similarity coefficient ranging from 0.59 to 0.87. Similar to RAPD results, ISSR analysis of the L. chinensis DNA samples showed a range of 0.70-0.93 similarity coefficients. The genetic distance between Hainan sample and Sichuan samples was the farthest, which is consistent with their geographic distance. Furthermore, the index of similarity coefficient between D. confinis and L. chinensis was 0.35-0.41 by RAPD and 0.38-0.48 by ISSR, indicating that these two species have significant genetic difference. This study reveals the high level of genetic differences between different litchi species or cultivars, and confirms the significance of the improved RAPD method in genetic characterization of organisms. Taken together, the improved RAPD combined with ISSR analysis can be used frequently for the genetic diversity, germplasm resources preservation, molecular-assisted breeding, and genetic characterization of various organisms.

Optimisation of Ultrasound-Assisted Extraction Conditions for Phenolic Content and Antioxidant Capacity from Euphorbia tirucalli Using Response Surface Methodology.

Euphorbia tirucalli (E. tirucalli) is now widely distributed around the world and is well known as a source of traditional medicine in many countries. This study aimed to utilise response surface methodology (RSM) to optimise ultrasonic-assisted extraction (UAE) conditions for total phenolic compounds (TPC) and antioxidant capacity from E. tirucalli leaf. The results showed that ultrasonic temperature, time and power effected TPC and antioxidant capacity; however, the effects varied. Ultrasonic power had the strongest influence on TPC; whereas ultrasonic temperature had the greatest impact on antioxidant capacity. Ultrasonic time had the least impact on both TPC and antioxidant capacity. The optimum UAE conditions were determined to be 50 °C, 90 min. and 200 W. Under these conditions, the E. tirucalli leaf extract yielded 2.93 mg GAE/g FW of TPC and exhibited potent antioxidant capacity. These conditions can be utilised for further isolation and purification of phenolic compounds from E. tirucalli leaf.

노니 뿌리 추출물의 pH 조건별 실크 염색성 고찰

Morinda citrifolia, commercially known as Noni, is a tree that grows widely throughout the Pacific, and is recognized as one of the most significant sources of traditional medicines among Pacific Island societies. The bark and the roots of the tree contain red and yellow pigments respectively, which are both used in the manufacture of dyes. Dyes from Noni are being used traditionally, to color clothing and fabrics. In this study, dyeing properties of Noni root extracts on silk fabrics at various pH conditions were investigated.

A Phytochemical, Pharmacological and Clinical Profile of Paederia foetida and P. scandens

Paederia foetida and P. scandens are two important and well explored Paederia species (Rubiaceae). P. foetida, which grows mainly in China, Bangladesh, India and Mauritius, has been used in folk medicine for the treatment of inflammation, piles, and diarrhea, while P. scandens is used to treat aches, jaundice, dysentery and dyspepsia as a folk medicine in the southern region of China, Vietnam, India and Japan. This review covers the comprehensive knowledge of the traditional medicinal uses, phytochemistry, pharmacology, toxicology and clinical studies of P. foetida and P. scandens. Phytochemical studies revealed the presence of iridoids, flavonoids, volatile oil, and other metabolites in these two species, which possess versatile bioactivities like antinociceptive, anti-inflammatory, antidiarrheal, antitussive and antitumor activities. An injection developed from P. scandens has been clinically used as an analgesic drug. P. foetida and P. scandens have emerged as a good source of traditional medicines. Available scientific references reveal that the biological properties of these two Paederia species have been evaluated by modem pharmacological studies. However, bioguided isolation of active constituent responsible for the medical uses, as well as study of their structure-activity relationship and mode of actions, is urgently needed.

Ameliorative effect of epigallocatechin gallate on sodium fluoride induced oxidative stress mediated metabolism in rat

Background: The present study was designed to examine the ant hyperglycemic and antihyperlipidemic effect of Epigallocatechin gallate (EGCG) on fluoride induced oxidative changes in the metabolisms of rats. Methods: The control group received the vehicles only. EGCG alone treated group received the EGCG orally at a dose of 40mg/kg in 10% tween 80 for 4 weeks. The Fl treated group received NaF (25 mg/kg) orally in saline for 4weeks. EGCG + Fl treated group received the EGCG at a dose of 40mg/kg/bw along with NaF. Results: The levels of serum SGOT, SGPT, ALP, ACP and plasma glucose, G6PD, cholesterol, triglycerides, free fatty acids, phospholipids, low density lipoprotein, very low density lipoprotein were significantly increased in fluoride intoxicated rats. Furthermore, a significant increase HMG CoA with decreased glycogen, hexokinase, total protein, albumins, high density lipoprotein, lipoprotein lipase, and lecithin cholesterol acyl transferase was observed in fluoride treated rats with altered histological changes. Pre-oral administration of EGCG at a dose of 40 mg/kg bw to fluoride intoxicated rats for a period of 28 days resulted in a significant reduction in the cholesterol, triglycerides, free fatty acids, phospholipids, low density lipoprotein, very low density lipoprotein, and elevation of high density lipoprotein with hexokinase. Moreover, lipoprotein lipase, lecithin cholesterol acyl transferase, total protein, and albumins were also increased with decreased serum markers and HMG CoA after EGCG treatment. The histological findings of liver also significantly improved with EGCG pretreatment. Conclusions: These results indicate that the natural dietary antioxidant EGCG showed significant protective effect against Fl- induced oxidative stress mediated metabolic alternations in rats. This may be due to its antihyperlipidemic and ant hyperglycemic property; it will provide an accessible and cheap traditional medicine source for treatment of Fl mediated environmental and occupational ailments. Keywords : EGCG, Fluoride, Metabolisms, Lipids, Anti-Hyperlipidemia ROS, Rat.

Anticonvulsant potential of some traditional medicinal plants

Epilepsy has now become the most serious brain disorder. A number of synthetic antiepileptic drugs are available in practice, however their effectiveness does not grip true with the entire population suffering from epilepsy. Traditional systems of medicine are popular in developing countries and most of the population relies on traditional medicines for their primary health care need. Medicinal plants to be an important source of traditional medicines. Various plants are used for the treatment of epilepsy in traditional system of medicines and various plants are yet to be scientifically investigated. Phyto-constituents have been the basis of treatment of human diseases including epilepsy. Herbal products are extensively used for the treatment of many diseases worldwide and where allopathic fails or has severe side effects. Psycho neural drugs are also have very serious side effects like physical dependence, tolerance, deterioration of cognitive function and effect on respiratory, digestive and immune system. So the treatments through herbal medicines are widely used across the world due to their wide applicability and therapeutic efficacy with least side effects, which in turn has accelerated the research regarding natural therapy. In this review we have summarized some herbal anti-epileptics.

Prospect ofPongamia pinnata(Karanja) in Bangladesh: A Sustainable Source of Liquid Fuel

Energy is the basic requirement for the existence of human being in today’s digital world. Indigenous energy of Bangladesh (especially natural gas and diesel) is basically used in power generation and depleting hastily to meet the increasing power demand. Therefore, special emphasis has been given to produce alternative liquid fuel worldwide to overcome the crisis of diesel.Pongamia pinnata(karanja) may be an emerging option for providing biooil for biodiesel production. Although karanja biooil has been used as a source of traditional medicines in Bangladesh, it can also be used for rural illumination. This paper outlines the medical and energy aspects ofPongamia pinnata. It has been assessed that Bangladesh can utilize about 128.95 PJ through Pongamia cultivation in unused lands. The paper reviews the potentiality ofPongamia pinnataas a source of biodiesel and its benefits in Bangladesh. The paper also revives that, about 0.52 million tons of biodiesel can be produced only utilizing the unused lands per year in sustainable basis as it reduces CO2, CO, HC, and NOxemission compared to pure diesel.

Diversity and Regeneration Aspects of Medicinal Plants at Devimane, Uttara Kannada District, Karnataka, Central Western Ghats

Diversity and Regeneration Aspects of Medicinal Plants at Devimane, Uttara Kannada District, Karnataka, Central Western Ghats Plants form an important source of traditional medicines for a major portion of population living in the tropical countries, since time immemorial. The Western Ghats of India is ranked among the 34 global biodiversity hotspots and is a storehouse of exemplarily rich biodiversity including numerous medicinal plants. However, excessive anthropogenic pressures on this ecosystem have resulted in severe depletion of natural resources and hence, this study was taken up to highlight the medicinal plant diversity and their regeneration aspects in Devimane area of Uttara Kannada district in Karnataka. Forest survey was carried out using a transect-based quadrat method revealing the presence of 348 plant species and about 40% of them are endemic to the Western Ghats-SriLanka region.

Foeniculum vulgare Mill: a review of its botany, phytochemistry, pharmacology, contemporary application, and toxicology.

Foeniculum vulgare Mill commonly called fennel has been used in traditional medicine for a wide range of ailments related to digestive, endocrine, reproductive, and respiratory systems. Additionally, it is also used as a galactagogue agent for lactating mothers. The review aims to gather the fragmented information available in the literature regarding morphology, ethnomedicinal applications, phytochemistry, pharmacology, and toxicology of Foeniculum vulgare. It also compiles available scientific evidence for the ethnobotanical claims and to identify gaps required to be filled by future research. Findings based on their traditional uses and scientific evaluation indicates that Foeniculum vulgare remains to be the most widely used herbal plant. It has been used for more than forty types of disorders. Phytochemical studies have shown the presence of numerous valuable compounds, such as volatile compounds, flavonoids, phenolic compounds, fatty acids, and amino acids. Compiled data indicate their efficacy in several in vitro and in vivo pharmacological properties such as antimicrobial, antiviral, anti-inflammatory, antimutagenic, antinociceptive, antipyretic, antispasmodic, antithrombotic, apoptotic, cardiovascular, chemomodulatory, antitumor, hepatoprotective, hypoglycemic, hypolipidemic, and memory enhancing property. Foeniculum vulgare has emerged as a good source of traditional medicine and it provides a noteworthy basis in pharmaceutical biology for the development/formulation of new drugs and future clinical uses.

Anticonvulsant potential of some medicinal plants and their beneficial properties

Epilepsy has now become the most serious brain disorder. A number of synthetic antiepileptic drugs are available in practice, however their effectiveness does not grip true with the entire population suffering from epilepsy. Traditional systems of medicine are popular in developing countries and most of the population relies on traditional medicines for their primary health care need. Medicinal plants to be an important source of traditional medicines. Various plants are used for the treatment of epilepsy in traditional system of medicines and various plants are yet to be scientifically investigated. Phytoconstituents have been the basis of treatment of human diseases including epilepsy. Herbal products are extensively used for the treatment of many diseases worldwide and where allopathic fails or has severe side effects. Psycho neural drugs are also have very serious side effects like physical dependence, tolerance, deterioration of cognitive function and effect on respiratory, digestive and immune system. So the treatments through herbal medicines are widely used across the world due to their wide applicability and therapeutic efficacy with least side effects, which in turn has accelerated the research regarding natural therapy. In this review we have summarized some herbal antiepileptics.

A strategy for trade monitoring and substitution of the organs of threatened animals

The use of threatened animals as a source of traditional medicines is accelerating the extinction of such species and imposes great challenges to animal conservation. In this study, we propose a feasible strategy for the conservation of threatened medicinal animals that combines trade monitoring and the search for substitutes. First, DNA barcoding provides a powerful technique for monitoring the trade of animal species, which helps in restricting the excessive use and illegal trade of such species. Second, pharmacological tests have been adopted to evaluate the biological equivalence of threatened and domestic animals; based on such testing, potential substitutes are recommended. Based on a review of threatened animal species and their substitutes, we find that the search for substitutes deserves special attention; however, this work is far from complete. These results may be of great value for the conservation of threatened animals and maintaining the heritage of traditional medicine.

Molecular cloning and development of RAPD-SCAR markers for Dimocarpus longan variety authentication

As an edible fruit and source of traditional medicine, D. longan is grown in most areas of Southern China. Identification of D. longan cultivars by using molecular markers is important genetically. In this study, we cloned fragments from improved randomly amplified polymorphic DNA (RAPD), and developed stably diagnostic sequence-characterized amplified region (SCAR) markers. The specific RAPD bands of D. longan cultivars from Guangxi, with size ranging from 500 bp to 900 bp were gel-purified, cloned and sequenced. Four clones named LY2-1, LY4-7, LY4-8 and LY5-2 were identified. In order to investigate whether the fragments were specific for the species, four pairs of SCAR primers were then designed. PCR amplifications were conducted to analyze 18 samples including different D. longan cultivars and other species. The specific bands with expected sizes were amplified in five D. longan samples but not in others. To identify and characterize the difference between D. longan and D. confinis, PCR amplifications were performed again. The specific bands with expected sizes were found in D. longan but not in D. confinis by SCAR markers LY2-1, LY4-7 and LY5-2, respectively. These results showed that our developed SCAR markers could be very useful as a specific D. longan variety authentication. Therefore, our study provides an effective and precise PCR-based diagnostic method and markers to identify D. longan species.