Multi-omics analysis reveals the evolutionary origin of diterpenoid alkaloid biosynthesis pathways in Aconitum.

Diterpenoid alkaloids (DAs) have been often utilized in clinical practice due to their analgesic and anti-inflammatory properties. Natural DAs are prevalent in the family Ranunculaceae, notably in the Aconitum genus. Nevertheless, the evolutionary origin of the biosynthesis pathway responsible for DA production remains unknown. In this study, we successfully assembled a high-quality, pseudochromosome-level genome of the DA-rich species Aconitum vilmorinianum (A. vilmorinianum) (5.76 Gb). An A. vilmorinianum-specific whole-genome duplication event was discovered using comparative genomic analysis, which may aid in the evolution of the DA biosynthesis pathway. We identified several genes involved in DA biosynthesis via integrated genomic, transcriptomic, and metabolomic analyses. These genes included enzymes encoding target ent-kaurene oxidases and aminotransferases, which facilitated the activation of diterpenes and insertion of nitrogen atoms into diterpene skeletons, thereby mediating the transformation of diterpenes into DAs. The divergence periods of these genes in A. vilmorinianum were further assessed, and it was shown that two major types of genes were involved in the establishment of the DA biosynthesis pathway. Our integrated analysis offers fresh insights into the evolutionary origin of DAs in A. vilmorinianum as well as suggestions for engineering the biosynthetic pathways to obtain desired DAs.

Correction: Structural diversity, bioactivities, and biosynthesis of natural diterpenoid alkaloids.

Correction for 'Structural diversity, bioactivities, and biosynthesis of natural diterpenoid alkaloids' by Yong Shen et al., Nat. Prod. Rep., 2020, 37, 763-796, https://doi.org/10.1039/D0NP00002G.

Aconitum Diterpenoid Alkaloid Profiling to Distinguish between the Official Traditional Chinese Medicine (TCM) Fuzi and Adulterant Species Using LC-qToF-MS with Chemometrics.

The lateral roots of Aconitum carmichaelii, known in Chinese as fuzi, are officially recognized as a materia medica in the Chinese Pharmacopoeia and used culinarily to prepare herbal soups. A strategy combining UPLC-qToF-MS analysis of A. carmichaelii and its intraspecies and interspecies chemometrics study was developed to examine the distribution of Aconitum marker metabolites. Four diterpenoid alkaloids were recognized to be important markers in fuzi, and another 15 markers were identified to differentiate A. carmichaelii from adulterant species. The detected fuzi markers, mesaconitine (47) and hypaconitine (51), are known to be the principal toxins in this herb, while fuziline (6) and benzoylmesaconine (25) are associated with its medicinal properties. Additional marker compounds have been detected in other Aconitum species that are useful for identifying adulteration. This study provides a useful resource for detecting traditional Chinese medicine (TCM) adulterants and assisting in the quality control of botanical products in TCM and beyond.

Structural diversity, bioactivities, and biosynthesis of natural diterpenoid alkaloids.

Covering: 2009 to 2018. Diterpenoid alkaloids, originating from the amination of natural tetracyclic diterpenes, are a diverse class of compounds having complex structural features with many stereocenters. The important pharmacological activities and structural complexity of the diterpenoid alkaloids have long interested scientists due to their medicinal uses, infamous toxicity, and unique biosynthesis. Since 2009, 373 diterpenoid alkaloids, assigned to 46 skeletons, have been isolated and identified from plants mostly in the Ranunculaceae family. The names, classes, molecular weight, molecular formula, NMR data, and plant sources of these diterpene alkaloids are collated here. This review will be a detailed update of the naturally occurring diterpene alkaloids reported from the plant kingdom from 2009-2018, providing an in-depth discussion of their diversity, biological activities, pharmacokinetics, toxicity, application, evolution, and biosynthesis.

Enriched networks 'nucleoside/nucleotide and ribonucleoside/ribonucleotide metabolic processes' and 'response to stimulus' potentially conferred to drought adaptation of the epiphytic orchid Dendrobium wangliangii.

Dendrobium wangliangii is an endangered and epiphytic orchid with tolerance to seasonally extreme arid conditions and occurs exclusively in the hot-dry valley area of southwestern China. To reveal its molecular basis responsible for ecological adaptation, large-scale transcriptome sequencing was performed using Illumina sequencing with pooled mRNA extracted from whole plants and pseudobulbs during drought and rainy seasons. Based on the target transcript selection, the differentially expressed genes were related to 8 well-known drought-tolerant categories, and to morphological traits in resistance to water stress including pseudobulbs and roots. Further gene ontology enrichment analysis revealed that 'nucleoside/nucleotide and ribonucleoside/ribonucleotide metabolic processes' and 'response to stimulus' were the two most important aspects in resistance to drought stress with respect to the whole plant. In addition, the difference in the number and category of differentially expressed genes in whole plant and stem suggested the involvement of genes specifically localized in the stem, such as GTP-binding protein, lipases, signaling related transcripts and those involved in the ATP metabolic process. The comprehensive analysis of the epiphytic orchid in response to water deprivation indicates that integral tactics lead to active adaptation as a basal defense response to drought stress by the endangered epiphyte, including the collaboration of metabolic processes, responses to a various stimulus and other candidate genes contribute to its extreme drought tolerance. Insights from this study can be further utilized to understand stress-responsive genes in other medicinally important species and to improve the drought tolerance of food crops.

Prognostic value of the age, creatinine, and ejection fraction score for non-infarct-related chronic total occlusion revascularization after primary percutaneous intervention in acute ST-elevation myocardial infarction patients: A retrospective study.

OBJECTIVE: It is not known if ACEF scores could evaluate the prognosis of recanalization of non-infarct-related coronary arteries (non-IRA) with chronic total occlusions (CTO) in patients who successfully underwent primary PCI. The objective of the current study was to assess the prognostic value of ACEF scores in acute ST-segment elevation myocardial infarction (STEMI) patients with non-IRA CTO after successful primary PCI. METHODS: There were 2952 STEMI patients who underwent successful primary PCI from January 2006 to December 2014 in our hospital, among them 377 patients had a non-IRA CTO lesion. The patients were divided into successful CTO-PCI group (n = 221) and failed/non-attempted CTO-PCI group (n = 156). Patients were stratified based on the ACEF tertiles. Primary end points measured in the current study were major adverse cardiac events (MACE) defined as the composite of all-cause death, nonfatal myocardial infarction, ischemia-driven coronary revascularization and hospitalization for heart failure at 1 year. RESULTS: The incidence of MACE, all-cause death and cardiac death were higher in the failed/non-attempted CTO-PCI group (P < 0.001). In the successful CTO-PCI group, the cumulative 1-year incidences of MACE and all-cause death were decreased compared to those in the failed/non-attempted CTO-PCI group (log-rank P < 0.001). The risk for MACE was reduced in the successful CTO-PCI group compared to the failed/non-attempted CTO-PCI group in patients with low and intermediate ACEF scores (log-rank P = 0.02). CONCLUSIONS: Successfully staged CTO-PCI could gain advantageous clinical outcomes in those patients with low or intermediate ACEF scores.

Identification of Potential Biomarkers from Aconitum carmichaelii, a Traditional Chinese Medicine, Using a Metabolomic Approach.

Despite their well-known toxicity, Aconitum species are important traditional medicines worldwide. Aconitum carmichaelii, known in Chinese as (fuzi), is an officially recognized traditional Chinese medicine with characteristic analgesic and anti-inflammatory activities, whose principal pharmacological ingredients are considered as aconitine-type diterpene alkaloids. Notwithstanding the long-recorded use of A. carmichaelii in traditional Chinese medicine, no single-entity aconitum alkaloid drug has been developed for clinical use. UPLC-Q-TOF-MS was used to investigate the marker compounds that can be used to differentiate A. carmichaelii from seven other Aconitum species collected in Yunnan Province. Nontargeted principle component analysis scores plots found that all the tested Aconitum species clustered into three distinct groups, and A. carmichaelii was significantly different chemically than the other seven species. Furthermore, the primary and lateral roots of A. carmichaelii also showed significant differences. Using orthogonal partial least squares discriminate analysis analysis, eight marker compounds were identified, including 14-acetylkarakoline, aconitine, carmichaeline, fuziline, hypaconitine, mesaconitine, neoline, and talatisamine. Four of these aconitum alkaloids, fuziline, hypaconitine, mesaconitine, and neoline, showed significant analgesic activity in a dose-dependent manner compared to the negative and positive controls. However, hypaconitine, mesaconitine, and neoline exhibited significant acute toxicity activity, while fuziline showed no acute toxicity in mice, suggesting the relative safety of this alkaloid. This study provides a good example of how to differentiate an authentic medicinal plant from common adulterants using a metabolomics approach, and to identify compounds that may be developed into new drugs.

The Role of Phyto-Melatonin and Related Metabolites in Response to Stress.

Plant hormone candidate melatonin has been widely studied in plants under various stress conditions, such as heat, cold, salt, drought, heavy metal, and pathogen attack. Under stress, melatonin usually accumulates sharply by modulating its biosynthesis and metabolic pathways. Beginning from the precursor tryptophan, four consecutive enzymes mediate the biosynthesis of tryptamine or 5-hydroxytryptophan, serotonin, N-acetylserotonin or 5-methoxytryptamine, and melatonin. Then, the compound is catabolized into 2-hydroxymelatonin, cyclic-3-hydroxymelatonin, and N¹-acetyl-N²-formyl-5-methoxyknuramine through 2-oxoglutarate-dependent dioxygenase catalysis or reaction with reactive oxygen species. As an ancient and powerful antioxidant, melatonin directly scavenges ROS induced by various stress conditions. Furthermore, it confreres stress tolerance by activating the plant's antioxidant system, alleviating photosynthesis inhibition, modulating transcription factors that are involved with stress resisting, and chelating and promoting the transport of heavy metals. Melatonin is even proven to defense against pathogen attacks for the plant by activating other stress-relevant hormones, like salicylic acid, ethylene, and jasmonic acid. Intriguingly, other precursors and metabolite molecules involved with melatonin also can increase stress tolerance for plant except for unconfirmed 5-methoxytryptamine, cyclic-3-hydroxymelatonin, and N¹-acetyl-N²-formyl-5-methoxyknuramine. Therefore, the precursors and metabolites locating at the whole biosynthesis and catabolism pathway of melatonin could contribute to plant stress resistance, thus providing a new perspective for promoting plant stress tolerance.

[Testing methods for seed quality of Bletilla striata].

In order to provide a basis for establishing seed testing rules and seed quality standard of Bletilla striata, the seed quality of B.striata from different producing area was measured referring to the Rules for Agricultural Seed Testing(GB/T 3543-1995).The results showed that the seeds of B.striata passed through 20-mesh sieve for purity analysis.The weight of seeds was measured by 1000-seed method and the water content was measured at the higher temperature (133±2) ℃ for 3 hours.The seeds were cultured on the wet filter paper at 30 ℃ for 4-20 days in light for germination testing.The method of testing seed viability was that seeds were dipped into 1% TTC solution for 7 hours at temperature of 40 ℃.

[Double mulching application for Panax notoginseng growing seedlings].

OBJECTIVE: In order to improve the irrigation for Panax notginseng growing seedlings, different mulching ways were carried out to investigate the effects of double mulching. METHOD: Field experiment was applied to study soil moisture, soil temperature and bulk density of different mulching ways while the germination rate and seedlings growth also were investigated. RESULT: Compared with the traditional single mulching with pine leaves or straw, double mulching using plastic film combined with pine leaves or straw could reduce 2/3 volumes of irrigation at the early seedling time Double mulching treatments didn't need to irrigate for 40 days from seeding to germination, and kept soil moisture and temperature steady at whole seedling time about 30% and 9.0-16.6 degrees C, respectively. The steady soil moisture and temperature benefited to resist late spring cold and germinate earlier while kept germination regularly, higher rate and seedlings quality. In contrast, single mulching using pine leaves or straw had poor soil moisture and temperature preserving, needed to irrigate every 12-day, meanwhile dropped the germination and booming time 14 days and 24-26 days, respectively, reduced germination rate about 11.3%-8.7%. However, single pine leaves mulching was better than straw mulching. In addition, though better effects of soil moisture and temperature preserving as well as earlier and higher rate of germination with single plastic films mulching had, some disadvantages had also been observed, such as daily soil temperature changed greatly, seedling bed soil hardened easily, more moss and weeds resulted difficulty in later management. CONCLUSION: To the purpose of saving water and labor as well as getting higher germination rate and seedlings quality, double mulching using plastic films combined pine leaves at the early time and single mulching removing plastic films at the later time is suggested to apply in the growing seedlings of P. notoginseng.

[Species and ecological control of disease on cultivated Gentiana rigescens in Yunnan].

OBJECTIVE: To find out variety of the fungal diseases of cultivated Gentiana rigescens and provide important basis for prevention. METHODS: The diseases were diagnosed based on field investigate, symptoms observation, pathogen isolation, determination the size of morphological and verification following the Koch's Postulate procedures. RESULTS: Anthracnose (Colletotrichum gloeosporioides), grey mould (Botrytis cinerea), brown spot (Alternaria tenuis), rust (Aecidiumpers), circular spot (Pestalotiopsis), leaf blight (Stemphylium, Ascochyta, Pleospora) and nematodes (Heterodera spp., Meloidogyne spp.) were found on Gentiana rigescens. Anthracnose was the first main disease, the diseased plant rate was over 40% and disease severity was 4 - 5 degree and second disease was rust, incidence of rate was less 10% and other diseases rate was not enough 2%. CONCLUSION: All these diseases on Gentiana rigescens are reported for the first time and Gentiana rigescens is the new host plant of the diseases.

Grafting: a potential method to reveal the differential accumulation mechanism of secondary metabolites.

Plant secondary metabolites make a great contribution to the agricultural and pharmaceutical industries. Their accumulation is determined by the integrated transport of target compounds and their biosynthesis-related RNA, protein, or DNA. However, it is hard to track the movement of these biomolecules in vivo. Grafting may be an ideal method to solve this problem. The differences in genetic and metabolic backgrounds between rootstock and scion, coupled with multiple omics approaches and other molecular tools, make it feasible to determine the movement of target compounds, RNAs, proteins, and DNAs. In this review, we will introduce methods of using the grafting technique, together with molecular biological tools, to reveal the differential accumulation mechanism of plant secondary metabolites at different levels. Details of the case of the transport of one diterpene alkaloid, fuziline, will be further illustrated to clarify how the specific accumulation model is shaped with the help of grafting and multiple molecular biological tools.

The uncoupled anterior and posterior spinal ligament tension (UAPLT) - An improvement to three-dimensional spring model of adolescent idiopathic scoliosis (AIS) pathogenesis.

The pathogenesis of Adolescent Idiopathic Scoliosis (AIS) remains unclear. Previous research proposed that ligament laxity is a clinical feature that can be easily overlooked in patients with AIS. We speculated a new hypothesis which is an improvement of our three-dimensional spring model hypothesis of AIS pathogenesis. The tethered string in the spring model stimulates the spinal ligament instead of spinal cord. Spinal overgrowth in the adolescent age leads to higher tension of posterior spinal ligament. And the ligament laxity leads to lower tension of anterior spinal ligament. This uncoupled anterior and posterior spinal ligament tension maybe the key cause of AIS.

Identification, Biological Activities and Biosynthetic Pathway of Dendrobium Alkaloids.

Dendrobium is a genus of flowering plants belonging to the Orchidaceae family with more than 1,400 species. Many Dendrobium species have been used as medicinal plants in several Asian countries for thousands of years. Alkaloids were reported as the major biological markers due to their complex chemical compositions and various types. In this review, we summarized the structural types of alkaloids, their pharmacological activities, as well as the mechanisms of biological activities. More than sixty alkaloids were isolated and identified from the Dendrobium genus. Moreover, the pharmacological effects of Dendrobium alkaloids as hepatic lipid and gluconeogenesis regulation, as neuroprotection, and as anti-tumor, anti-inflammatory, anti-diabetes, and anti-virus factors were described. Besides, the total chemical synthesis of dendrobine is provided, while the biosynthetic pathway of dendrobine has been proposed based on the functions of associated genes. For applications of these invaluable herbs, more researches on the extraction of biological markers from compounds are needed. Further confirmation of the proposed biosynthetic pathways is anticipated as well.

Clerodane diterpenoids and prenylated flavonoids from Dodonaea viscosa.

Repeated column chromatography of the EtOAc-soluble fraction of the aerial parts of Dodonaea viscosa led to the isolation of two new modified clerodanes, methyl dodovisate A (1) and methyl dodovisate B (2), two new prenylated flavonoids, 5,7,4'-trihydroxy-3',5'-di(3-methylbut-2-enyl)-3,6-dimethoxyflavone (10) and 5,7,4'-trihydroxy-3'-(4-hydroxy-3-methylbutyl)-5'-(3-methylbut-2-enyl)-3,6-dimethoxyflavone (11), together with eight known compounds, dodonic acid (3), hautriwaic acid (4), hautriwaic lactone (5), (+)-hardwickiic acid (6), 5alpha-hydroxy-1,2-dehydro-5,10-dihydroprintzianic acid methyl ester (7), strictic acid (8), dodonolide (9), and aliarin (12). The structures of the new compounds were elucidated by spectroscopic data analysis. Compounds 1-9 and 11 were evaluated on larvicidal activity against the fourth-instar larvae of Aedes albopictus and Culex pipens quinquefasciatus.

Sesquiterpenoids from Pilea cavaleriei subsp. crenata.

Three new humulane-type sesquiterpenes, 8-O-(p-coumaroyl)-5beta-hydroperoxy-1(10)E,4(15)-humuladien-8alpha-ol (1), 8-O-(3-nitro-p-coumaroyl)-1(10)E,4(15)-humuladien-5beta,8alpha-diol (2) and 8-O-(p-coumaroyl)-1(10)E,4(5)E-humuladien-8-ol (3), and a new copaborneol derivative, 1-O-p-coumaroyl-copaborneol (4), have been isolated from the methanol extract of Pileacavaleriei Lévl. subsp. crenata C. J. Chen. Their structures were elucidated using spectroscopic methods. Cytotoxic and antimicrobial activities of the isolated compounds were evaluated.

Pyrrolidinoindoline alkaloids from Selaginella moellendorfii.

Eight new pyrrolidinoindoline alkaloids (1-8) were isolated from the whole plant of Selaginella moellendorfii. Their structures were determined by mass spectrometry, 1D and 2D NMR spectroscopy, and chemical interconversions. These alkaloids have a 3-carboxybut-2-enyl group at C-3a and two methyl groups at N-8. The possible biogenetic route from selaginellic acid (1) to neoselaginellic acid (6) was postulated and chemically mimicked. Tautomerization between 6 and 6a was observed. Selected compounds were evaluated for antibacterial, cytotoxic, and acetylcholinesterase inhibitory activities.

Chemical constituents in roots of Osbeckia opipara.

OBJECTIVE: To study the chemical constituents of the roots of Osbeckia opipara. METHOD: Repeated column chromatography over silica gel, RP-18 and Sephadex LH-20, and preparative thin layer chromatography(PTLC) were used to isolate the compounds, whose structures were determined by spectroscopic methods by direct comparing spectral data with those reported references. RESULT: From the MeOH extract of the roots O. opipara, twelve compounds were isolated and identified as follows: lasiodiplodin (1) , de-O-methyllasiodiplodin (2), 2, 3- dihydro-2-hydroxy-2, 4-dimethyl-5-trans-propenylfuran-3-one (3), integracin (4), 5alpha, 8alpha-epidioxy-(22E, 24R)-ergosta-6, 22-dien-3beta-ol (5), 3, 3', 4'-tri-O-methylellagic acid (6), 5-hydroxymethyl furaldehyde (7), vomifolio (8) , betulintic acid (9), 2alpha-hydroxyursolic acid (10), (24R)-stigmast-4-ene-3-one (11), and eugenitin (12). CONCLUSION: Compounds 1-12 were isolated from O. opipara for the first time.

Jaboticabin and Related Polyphenols from Jaboticaba ( Myrciaria cauliflora) with Anti-inflammatory Activity for Chronic Obstructive Pulmonary Disease.

Myrciaria cauliflora (jaboticaba) is an edible fruit common in Brazil that has been used for treating respiratory diseases, including chronic tonsillitis and asthma. This study explores the distribution of an anti-inflammatory depside, jaboticabin, in different parts of the jaboticaba plant as well as major polyphenols from the wood of jaboticaba, some with biological activity similar to jaboticabin. The peel of the fruit was found to be the major source of jaboticabin. This is the first phytochemical study of the wood of M. cauliflora. The antioxidant-activity-guided fractionation strategy successfully identified 3,3'-dimethylellagic acid-4- O-sulfate from jaboticaba wood. This ellagic acid derivative, in a manner similar to jaboticabin, showed antiradical activity and inhibited the production of the chemokine interleukin-8 after treating the human small airway epithelial cells with cigarette smoke extract. The human intestinal Caco-2 cell studies demonstrated the jaboticabin transport in vitro. The polyphenols, jaboticabin and 3,3'-dimethyellagic acid-4- O-sulfate, from jaboticaba were both found to exhibit anti-inflammatory activities, thus suggesting the potential use of these compounds or even the fruits themselves for chronic obstructive pulmonary disease.

Chemical constituents of Aeschynanthus bracteatus and their weak anti-inflammatory activities.

Chemical examination of the EtOAc extract from the aerial parts of Aeschynanthus bracteatus led to isolation of four phenylethanol glycosides, aeschynanthosides A-D (1-4), and 55 known constituents, including 8 phenylethanoids, 23 phenols, 5 lignans, 7 flavonoids, 9 terpenoids, and 4 others. Their structures were elucidated mainly by detailed spectroscopic studies and comparison with published data. All 59 compounds were isolated for the first time from an Aeschynanthus species. The isolates were also tested for inhibitory activities against LPS-induced NO production in RAW 264.7 macrophages. Aeschynanthoside D (4) and naringenin (41), within the concentration arrange tested (50-100 microg/mL), showed very weak dose-dependent effects with the inhibition rate of 24.2%, 35.4%, 66.0%, and 9.5%, 40.1%, 65.0%, respectively, relative to positive controls.