Comparative transcriptome analysis and simple sequence repeat marker development for two closely related Isodon species used as ‘Xihuangcao’ herbs

Purpose: To facilitate the molecular identification of original plants, resolve taxonomic problems and identify standards for ‘Xihuangcao’-based products on the market.Methods: A transcriptomic analysis of two closely related species, i.e., Isodon serra (Maxim.) (IS) and I. lophanthoides (Buch.-Ham. ex D. Don) Hara, was conducted by using the Illumina HiSeq 2500 platform, and expressed sequence tag-derived simple sequence repeat (EST-SSR) markers were developed based on these transcriptomes.Results: In total, 149,650 and 103,221 contigs were obtained, with N50 values of 1,400 and 1,516, from the IS and I. lophanthoides RNA-Seq datasets, respectively. These contigs were clustered into 107,777 and 68,220 unigenes, which were functionally annotated to identify the genes involved in therapeutic components. In total, 14,138 and 11,756 EST-SSR motifs were identified, and of these motifs, 7,453 and 6,428 were used to design primers for IS and I. lophanthoides, respectively. After PCR verification and fluorescence-based genotyping, 24 SSR markers with bright bands, high polymorphism, and single amplification were obtained and used to identify closely related Isodon species/varieties.Conclusion: These data could help herbal scientists identify high-quality herbal plants and provide a reference for genetic improvement and population genetic and phylogenetic studies investigating ‘Xihuangcao’ herbs.Keywords: Xihuangcao, Transcriptome, EST-SSR, Molecular marker

Analysis of genetic variability in patchouli cultivars (Pogostemon cablin Benth.) by using RAPD Markers

The genetic relationships among patchouli cultivars were determined by using Random Amplified Polymorphic DNA (RAPD) technology. Among 45 decamer random primers used for PCR reactions, 10 primers showed reproducible results. Out of 98 amplification products recorded, 16.7 per cent were monomorphic and 83.3 per cent were polymorphic. The highest dissimilarity (7.35) was detected between KSM 4 and 5 and the least 3.61 between KSM 2 and 3. Dendrogram constructed by cluster analysis of RAPD markers using Unweighted Pair Group Method of Arithmetic Averages  (UPGMA) produced two major clusters ‘A’ and ‘B’. Cluster ‘A’ consisted of five cultivars that segregated into two sub clusters ‘A1’ and ‘A2’. Overall, RAPD analysis revealed the existence of considerable genetic variations in patchouli cultivars. This information regarding genetic variability at the molecular level could be used to identify and develop genetically unique germplasm that complements existing cultivars

Isodon Diterpenoids, Derivatives and their Pharmacological Potentials-A Review

The searches of various literatures have shown the genus Isodon to be a source of different compounds with interesting biological activities. The genus has provided many efficacious herbal medicines that are used in various countries including China; therefore, it has become the centre of attention to phytochemistry and pharmacology researchers. There are many reports on chemical and biological aspects of Isodon species, especially in China and other parts of the world; however, reports on African Isodon species are scanty. Since the literature indicates the genus to be rich in diterpenoids with potential therapeutic activities as revealed herein, with African species waiting to be explored, it is the responsibility of the phytochemists and pharmacologists to fill this knowledge gap. Herein, ethnomedicinal uses of some of Isodon plants in various traditional medicine systems, phytochemistry of the genus from 2016 to date, synthesis of Isodon diterpenoids and derivatives are discussed. Keywords: Isodon diterpenoids; Natural Products; Isodon phytochemistry; Herbal Medicines; Diterpenoids synthesis

Chemical Authentication of Botanical Ingredients: A Review of Commercial Herbal Products.

Chemical methods are the most important and widely used traditional plant identification techniques recommended by national and international pharmacopoeias. We have reviewed the successful use of different chemical methods for the botanical authentication of 2,386 commercial herbal products, sold in 37 countries spread over six continents. The majority of the analyzed products were reported to be authentic (73%) but more than a quarter proved to be adulterated (27%). At a national level, the number of products and the adulteration proportions varied very widely. Yet, the adulteration reported for the four countries, from which more than 100 commercial products were purchased and their botanical ingredients chemically authenticated, was 37% (United Kingdom), 31% (Italy), 27% (United States), and 21% (China). Simple or hyphenated chemical analytical techniques have identified the total absence of labeled botanical ingredients, substitution with closely related or unrelated species, the use of biological filler material, and the hidden presence of regulated, forbidden or allergenic species. Additionally, affecting the safety and efficacy of the commercial herbal products, other low quality aspects were reported: considerable variability of the labeled metabolic profile and/or phytochemical content, significant product-to-product variation of botanical ingredients or even between batches by the same manufacturer, and misleading quality and quantity label claims. Choosing an appropriate chemical technique can be the only possibility for assessing the botanical authenticity of samples which have lost their diagnostic microscopic characteristics or were processed so that DNA cannot be adequately recovered

Microscopic Authentication of Commercial Herbal Products in the Globalized Market: Potential and Limitations

Herbal products are marketed and used around the globe for their claimed or expected health benefits, but their increasing demand has resulted in a proportionally increase of their accidental contamination or intentional adulteration, as already confirmed with DNA-based methods. Microscopy is a traditional pharmacopoeial method used for plant identification and we systematically searched for peer-reviewed publications to document its potential and limitations to authenticate herbal medicines and food supplements commercially available on the global market. The overall authenticity of 508 microscopically authenticated herbal products, sold in 13 countries, was 59%, while the rest of 41% were found to be adulterated. This problem was extending over all continents. At the national level, there were conspicuous differences, even between neighboring countries. These microscopically authenticated commercial herbal products confirm that different magnifying instruments can be used to authenticate crude or processed herbal products traded in the global marketplace. The reviewed publications report the successful use of different magnifying instruments, single or in combinations with a second one, with or without a chemical or DNA-based technique. Microscopy is therefore a rapid and cost-efficient method, and can cope with mixtures and impurities. However, it has limited applicability for highly processed samples. Microscopic authentication of commercial herbal products will therefore contribute to raise public awareness for the extent of adulteration and the need to safeguard consumer safety against the challenges of globalization

Anti-Inflammatory ent-Kaurane Diterpenoids from Isodon serra

Ten new ent-kaurane diterpenoids, including two pairs of epimers 1/2 and 4/5 and a 6,7-seco-ent-kauranoid 10, were obtained from the aerial parts of Isodon serra. The structures of the new compounds were confirmed by extensive spectroscopic methods and electronic circular dichroism (ECD) data analysis. An anti-inflammatory assay was applied to evaluate their nitric oxide (NO) inhibitory activities by using LPS-stimulated BV-2 cells. Compounds 1 and 9 exhibited notable NO production inhibition with IC50 values of 15.6 and 7.3 μM, respectively. Moreover, the interactions of some bioactive diterpenoids with inducible nitric oxide synthase (iNOS) were explored by employing molecular docking studies.https://pubs.acs.org/journal/jnprdf2021-09-29hj2021Plant Production and Soil Scienc

Pollen Morphology of Chinese Medicine Xihuangcao and Its Related Species

用光学显微镜和扫描电镜对中药溪黄草４种基源植物的花粉形态进行了观察和比较研究．结果表明：光学显微镜下该四种植物的花粉形态相似，赤道面观长球形至扁球形，极面观６裂圆形，具６沟，外壁具粗网状纹饰，而扫描电镜下４种花粉在外壁纹饰、萌发沟等方面存在着明显差异，这些差异可为近缘种的鉴定以及该类生药的鉴定提供依据．Pollen morphology of four original plants of Chinese Medicine Xihuangcao was examined by light microscope (LM) and scanning electron microscope (SEM ) for the first time. Pollen grains are spheroidal or subspheroidal,prolate or subprolate in shape and 6-lobed-circular in polar view, 6-loppate; The ornamentation of exine is coarsely reticulate. Under SEM, apparent differences can be observed about ornamentation of pollen exine which may be regarded as new basis to identify and classify Chinese Medicine Xihuangcao and its Related Species.广东省自然科学基金!（940474）;; 国家自然科学基金资助项目!（39870461

DiFFerentiation of Original Plants of Xihuangcao by Using Random AmpliFied Polymorphic DNA (RAPD) Markers

采用rAPd技术对中药溪黄草类4种原植物进行鉴定研究，结果表明rAPd技术是一种有效的中药鉴定方法，对药材的“地道性”研究有重要意义Four original plants of Xihuangcao have been diFFerentiated From each other by random ampliFied polymorphic DNA polymorphism.The results show that RAPD technique is an eFFective method For identiFication and quality study of Chinese medicinal materials.广东省自然科学基金;国家杰出青年基