https://link.springer.com/content/pdf/10.1186/s43141-019-0011-6.pdf

Molecular analysis of genetic diversity in population of Silybum marianum (L.) Gaertn in Egypt.

Substantial variation in phenotypic traits and ISSR fingerprinting was observed among twenty populations of Achillea fragrantissima (Forssk.) Sch. Bip. in Egypt. Such variation was reflected in the clustering of the examined populations into two major groups: one representing populations in the mountainous area of South Sinai and the other comprising populations growing at low elevations in the middle of Sinai and the desert west of the Suez Canal from Suez in the east to Cairo in the west. Five populations in the eastern part of Sinai near Nuwieba and Taba on the Gulf of Aqaba were loosely assigned to the first group. The populations growing at high elevations in South Sinai, under lower temperature and higher humidity, were characterized by a higher number of total and polymorphic ISSR markers compared with other populations. Unique ISSR markers were more often observed in the fingerprinting of seven populations including five populations growing in the high mountains of Saint Catherine in South Sinai and two populations growing at low elevations but at Wadi Hof south-east of Cairo. Interestingly, unique bands were found in the populations that possessed traits associated with larger plant size and seed yield as well as better vigour. These are important criteria for the selection of A. fragrantissima populations for conservation and sustainable commercial use.

Genetic diversity of Achillea fragrantissima in Egypt inferred from phenotypic variations and ISSR markers associated with traits of plant size and seed yield

Medicinal effects of Crepidiastrum denticulatum have been previously reported. However, the genomic resources of this species and its applications have not been studied. In this study, based on the next generation sequencing method (Miseq sequencing system), we characterize the chloroplast genome of C. denticulatum which contains a large single copy (84,112 bp) and a small single copy (18,519 bp), separated by two inverted repeat regions (25,074 bp). This genome consists of 80 protein-coding gene, 30 tRNAs, and four rRNAs. Notably, the trnT_GGU is pseudogenized because of a small insertion within the coding region. Comparative genomic analysis reveals a high similarity among Asteraceae taxa. However, the junctions between LSC, SSC, and IRs locate in different positions within rps19 and ycf1 among examined species. Also, we describe a newly developed single nucleotide polymorphism (SNP) marker for C. denticulatum based on amplification‐refractory mutation system (ARMS) technique. The markers, inferred from SNP in rbcL and matK genes, show effectiveness to recognize C. denticulatum from other related taxa through simple PCR protocol. The chloroplast genome-based molecular markers are effective to distinguish a potentially medicinal species, C. denticulatum, from other related taxa. Additionally, the complete chloroplast genome of C. denticulatum provides initial genomic data for further studies on phylogenomics, population genetics, and evolutionary history of Crepidiastrum as well as other taxa in Asteraceae.

The newly developed single nucleotide polymorphism (SNP) markers for a potentially medicinal plant, Crepidiastrum denticulatum (Asteraceae), inferred from complete chloroplast genome data.

Molecular pharmacognosy is based on molecular identification technologies and has provided an identification basis of crude drugs at the gene level. Molecular pharmacognosy includes the systematic assortment of varieties of traditional herbal medicine and Chinese herbs, and studies quality, standardization, conservation, and diversity of medicinal plants. The molecular pharmacognosy in twenty-first century embraces many items from the study of molecular taxonomy, phylogenetic evolution of medicinal plants and animals, conservation of endangered species, molecular identification of medicinal raw materials, controlling metabolic pathways and biosynthetic regulation of secondary metabolites in plants, conservation of biodiversity, genetic engineering, tissue culture technology, production of pollution-free medicinal plants, structure-activity relationships with a drug potential, etc. to genetic engineering technology based on molecular cloning and the related tissue culture technology, especially molecular marker technology based on PCR. Molecular pharmacognosy is a rising discipline combining molecular biology and pharmacognosy, and its development depends on systems biology with reference to genomics, proteomics, and metabolomics, and also involved methods of modern biotechnology. Molecular biology has become vital to drug discovery from medicinal plants, through the determination and accomplishment of appropriate screening assays directed toward physiologically relevant molecular targets. DNA markers based tools for molecular identification of traditional medicinal materials are considered more reliable for authentication of herbal materials.

Molecular Pharmacognosy—A New Borderline Discipline Between Molecular Biology and Pharmacognosy

Morphological characterization of diploid and triploid Acorus calamus (Acoraceae) from southern Western Siberia, parthenocarpy in sterile plants and occurrence of aneuploidy

The demand for herbal medicines is increasing day-to-day because of their widebiological activity, higher safety margin than synthetic drugs. Hence, it is important tocharacterize the medicinal herbs. For the characterization of medicinal plants it is necessaryto develop the sensitive and effective technology. Molecular markers can be employed forcharacterization, since the progress made in genetic markers has been exciting. DNA-basedmolecular markers have acted as very useful tools in various fields like taxonomy,physiology, embryology, plant breeding, ecology, genetic engineering etc. DNA basedmarkers have their applications in fingerprinting genotypes, determining the seed purityand in phylogenetic analysis by which the conservation of the plant can be made easy. Theinnovation of Polymerase Chain Reaction (PCR) made the development of DNA basedmarkers easier. Extensive research on molecular markers is in progress in many researchinstitutes all over the world. DNA-based molecular markers have a great utility in the druganalysis and widely used for the characterization of medicinally important plant species.This review is an attempt to evaluate critically the role of different DNA based markers inthe characterization of medicinal plants.

Molecular markers in characterization of medicinal plants: An overview

The use of plants and plant products for medicine is being practiced from time immemorial. The advent of herbal drug technology has led to the production of medicines and pharmaceutical products from herbs. Secondary metabolites from plants are important economically as drugs, fragrances, pigments, food additives and pesticides. In order to protect consumers from adulteration and for conservation of the plants; authentication and identification of medicinal plants is essential. Traditionally, morphological, phytochemical and analytical techniques are used. In recent times molecular marker based methods are used widely. With the advent of PCR based techniques minute quantities of DNA in order of nanograms are sufficient for molecular analysis. These techniques are more advantageous than the classical morphology based or chemical and analytical methods. Recently the nontranscribed sequence (NTS) region of 5S rRNA has been employed for studying intra specific discrimination. In this short review an attempt has been made to study the use of 5S -rRNA NTS region for molecular fingerprinting in plants.

Molecular characterization of medicinal and aromatic plants by 5S rRNA NTS and PCR RFLP- A mini review

Acorus calamus L. (Family: Acoraceae) is a well-known traditional, endangered, medicinal and aromatic plant mainly found in India and China. The plant is also widely used in industrial, pharmaceutical and food industries. In the present study, 20 different accessions of Indian A. calamus were subjected to the study of genetic diversity (RAPD), and cytogenetic and phytochemical (β-asarone) analysis. For RAPD analysis, 9 primers were chosen, which generated 107 DNA fragments. The average percentage of polymorphism was recorded to be 67.23%. The primer OPA 12 showed the highest (100%) polymorphism, whereas the lowest (38.2%) polymorphism was observed for the primer OPBB 6. The polymorphism information content (PIC) values ranged 0.44 (OPA 7) to 0.18 (OPA11), while marker index (MI) values ranged 4.74 (OPA 7) to 0.36 (OPA 11). A dendogram was constructed by UPGMA method and the robustness of the tree was confirmed by bootstrap analysis with 1000 pseudo samples. For cytogenetic analysis, the 20 A. calamus accessions were screened for their ploidy status. The accessions were found to be either diploid or triploids. The phytochemical analysis of β-asarone content was determined through by HPLC method. The β-asarone concentration varied in the range of 2.2 to 7.2 mg/100 mg. The results of present study indicated the presence of low level of polymorphism among the A. calamus accessions of South India and North-East India. The phytochemical and cytogenetic analysis revealed that both diploid and triploid have low concentration of β-asarone irrespective of their geographical location.

http://nopr.niscair.res.in/bitstream/123456789/41025/1/IJBT%2015%284%29%20560-567.pdf

Genetic diversity analysis and chemical profiling of Indian Acorus calamus accessions from South and North-East India

Acorus calamus is a well-known medicinal and aromatic plant used for many industrial and pharmaceutical purposes. In our current study, twenty different accessions of Acorus calamus collected from North-East India and South India have been studied and an attempt has been made to classify them according to their ploidy level by cytogenetic analysis and to correlate them via PCR-RFLP technique for the NonTranscribed Spacer (NTS) region of the 5S-rRNA gene. β-asarone which is the major phyto-chemical found in Acorus sp. when present in higher concentrations was found to be carcinogenic. The scope of this study is to identify diploid Acorus calamus accessions as they are reported to have less concentration of β-asarone from those of triploids. Cytoyping was performed by staining of root metaphase chromosomes. For PCR-RFLP analysis, the NTS region of 5S rRNA gene was amplified by using primers, designed from the coding regions of 5S rRNA gene. The PCR products were digested with EcoRI restriction enzyme. Triploid accessions showed restriction digestion at 586 bp resulting in an additional fragment of 127 bp, which is absent in diploids. Ten diploid accessions and 10 triploid accessions were identified by cytogenetic technique and PCR-RFLP method. The sequences of triploid and diploid were analyzed using bioinformatics tools.

/pdf/an-alternative-method-for-cytotype-determination-in-acorus-1h1k59lmvm.pdf

Tharachand C

Papers

Molecular markers in characterization of medicinal plants: An overview

Molecular characterization of medicinal and aromatic plants by 5S rRNA NTS and PCR RFLP- A mini review

Genetic diversity analysis and chemical profiling of Indian Acorus calamus accessions from South and North-East India

An Alternative Method for Cytotype Determination in Acorus calamus L., Accessions by PCR- RFLP Technique