Cannabis sativa: origin and history, glandular trichome development, and cannabinoid biosynthesis

Is Cannabis a boon or bane?Cannabis sativa has long been a versatile crop for fiber extraction(industrial hemp),traditional Chinese medicine(hemp seeds),and recreational drugs(marijuana).Cannabis faced global prohibition in the twentieth century because of the psychoactive properties of △^(9)-tetrahydrocannabinol;however,recently,the perspective has changed with the recognition of additional therapeutic values,particularly the pharmacological potential of cannabidiol.A comprehensive understanding of the underlying mechanism of cannabinoid biosynthesis is necessary to cultivate and promote globally the medicinal application of Cannabis resources.Here,we comprehensively review the historical usage of Cannabis,biosynthesis of trichome-specific cannabinoids,regulatory network of trichome development,and synthetic biology of cannabinoids.This review provides valuable insights into the efficient biosynthesis and green production of cannabinoids,and the development and utilization of novel Cannabis varieties.

The genome sequence of star fruit(Averrhoa carambola)

Oxalidaceae is one of the most important plant families in horticulture,and its key commercially relevant genus,Averrhoa,has diverse growth habits and fruit types.Here,we describe the assembly of a high-quality chromosomescale genome sequence for Averrhoa carambola(star fruit).Ks distribution analysis showed that A.carambola underwent a whole-genome triplication event,i.e.,the gamma event shared by most eudicots.Comparisons between A.carambola and other angiosperms also permitted the generation of Oxalidaceae gene annotations.We identified unique gene families and analyzed gene family expansion and contraction.This analysis revealed significant changes in MADS-box gene family content,which might be related to the cauliflory of A.carambola.In addition,we identified and analyzed a total of 204 nucleotide-binding site,leucine-rich repeat receptor(NLR)genes and 58 WRKY genes in the genome,which may be related to the defense response.Our results provide insights into the origin,evolution and diversification of star fruit.

A Research on the Behavior of a Polyurethane Polymer Waterproof Material Used in Bridge Geotechnical Applications

Polyurethane is enjoying a widespread use as a polymer-based waterproof material in civil engineering In the present study we consider a temperature-sensitive waterproof and moisture-permeable polyurethane material(PTPE-PU)characterized by one or more phase transition temperatures(critical temperatures).Near the critical temperature,the waterproof and moisture permeability of polyurethane undergo abrupt changes.The related stability,thermal performance,water resistance,hydrostatic pressure,and moisture permeability are investigated here considering a PTPE-PU traditionally used in bridge geotechnical engineering.The results show that the moisture permeability of the coated bridge rock and soil undergo sudden variations near the crystallization and melting temperature of the soft segment.The moisture permeability is 3000 g/(m^(2)d).The hydrostatic pressure of the coated bridge rock and soil is 3.5 kPa.

Crocus genome reveals the evolutionary origin of crocin biosynthesis

Crocus sativus (saffron) is a globally autumn-flowering plant, and its stigmas are the most expensive spice and valuable herb medicine. Crocus specialized metabolites, crocins, are biosynthesized in distant species, Gardenia (eudicot) and Crocus (monocot), and the evolution of crocin biosynthesis remains poorly understood. With the chromosome-level Crocus genome assembly, we revealed that two rounds of lineage-specific whole genome triplication occurred, contributing important roles in the production of carotenoids and apocarotenoids. According to the kingdom-wide identification, phylogenetic analysis, and functional assays of carotenoid cleavage dioxygenases (CCDs), we deduced that the duplication, site positive selection, and neofunctionalization of Crocus-specific CCD2 from CCD1 members are responsible for the crocin biosynthesis. In addition, site mutation of CsCCD2 revealed the key amino acids, including I143, L146, R161, E181, T259, and S292 related to the catalytic activity of zeaxanthin cleavage. Our study provides important insights into the origin and evolution of plant specialized metabolites, which are derived by duplication events of biosynthetic genes.

Genome Analysis of the Ancient Tracheophyte Selaginella tama#scina Reveals Evolutionary Features Relevant to the Acquisition of Desiccation Tolerance

Resurrection plants, which are the ＂gifts＂ of natural evolution, are ideal models for studying the genetic basis of plant desiccation tolerance. Here, we report a high-quality genome assembly of 301 Mb for the diploid spike moss Selaginella tamariscina, a primitive vascular resurrection plant. We predicated 27 761 protein-coding genes from the assembled S. tarnariscina genome, 11.38% （2363） of which showed signifi- cant expression changes in response to desiccation. Approximately 60.58% of the S. tamariscina genome was annotated as repetitive DNA, which is an almost 2-fold increase of that in the genome of desiccation- sensitive Selaginella moellendorffii. Genomic and transcriptomic analyses highlight the unique evolution and complex regulations of the desiccation response in S. tamariscina, including species-specific expan- sion of the oleosin and pentatricopeptide repeat gene families, unique genes and pathways for reactive oxygen species generation and scavenging, and enhanced abscisic acid （ABA） biosynthesis and potentially distinct regulation of ABA signaling and response. Comparative analysis of chloroplast genomes of several Selaginella species revealed a unique structural rearrangement and the complete loss of chloroplast NAD（P） H dehydrogenase （NDH） genes in S. tamariscina, suggesting a link between the absence of the NDH com- plex and desiccation tolerance. Taken together, our comparative genomic and transcriptomic analyses reveal common and species-specific desiccation tolerance strategies in S. tamariscina, providing signifi- cant insights into the desiccation tolerance mechanism and the evolution of resurrection plants.

西红花苷生源途径解析及其合成生物学研究进展

天然产物是新药研发的重要资源,具有显著的抗疟疾、抗肿瘤、抗病毒感染等作用.珍稀名贵中药西红花的活性成分为西红花苷,属于脱辅基类胡萝卜素类化合物,具有保护中枢神经系统及预防心脑血管疾病的药理活性.西红花苷资源匮乏、价格高昂,极大限制其药物开发及临床应用.本文从西红花苷的化学表征、生物合成途径解析及合成生物学研究进行综述,展示了西红花苷的光谱学和质谱学特征,阐述其完整生物合成途径的解析,介绍了利用微生物细胞工厂生产5种西红花苷的最新突破.据此,进一步提出开展不同物种西红花苷生物合成途径关键酶的进化机制、合成生物学关键技术探索和条件优化等研究,以推动西红花苷工业化生产及新药研发,为西红花苷来源的中药资源可持续发展提供理论基础和方法指导.

Large-scale analysis of small RNAs derived from traditional Chinese herbs in human tissues

Plant-derived microRNAs have recently been reported to function in human blood and tissues. Controversy was immediately raised due to possible contamination and the lack of large sample sizes. Here, we report thousands of unique small RNAs derived from traditional Chinese medicine(TCM) herbs found in human blood cells and mouse lung tissues using a large-scale analysis.We extracted small RNAs from decoctions of 10 TCM plants(Ban Zhi Lian, Chai Hu, Chuan Xin Lian, Di Ding Zi Jin, Huang Qin, Jin Yin Hua, Lian Qiao, Pu Gong Ying, Xia Ku Cao, and Yu Xing Cao) and obtained millions of RNA sequences from each herb. We also obtained RNA-Seq data from the blood cells of humans who consumed herbal decoctions and from the lung tissues of mice administered RNAs from herbal decoctions via oral gavage. We identified thousands of unique small RNA sequences in human blood cells and mouse lung tissues. Some of these identified small RNAs from Chuan Xin Lian and Hong Jing Tian could be mapped to the genomes of the herbs, confirming their TCM plant origin. Small RNAs derived from herbs regulate mammalian gene expression in a sequence-specific manner, and thus are a superior novel class of herbal drug components that hold great potential as oral gene-targeted therapeutics, highlighting the important role of herbgenomics in their development.

Biomonitoring for traditional herbal medicinal products using DNA metabarcoding and single molecule, real-time sequencing

Global concerns have been paid to the potential hazard of traditional herbal medicinal products(THMPs). Substandard and counterfeit THMPs, including traditional Chinese patent medicine, health foods, dietary supplements, etc. are potential threats to public health. Recent marketplace studies using DNA barcoding have determined that the current quality control methods are not sufficient for ensuring the presence of authentic herbal ingredients and detection of contaminants/adulterants. An efficient biomonitoring method for THMPs is of great needed. Herein, metabarcoding and single-molecule, realtime(SMRT) sequencing were used to detect the multiple ingredients in Jiuwei Qianghuo Wan(JWQHW), a classical herbal prescription widely used in China for the last 800 years. Reference experimental mixtures and commercial JWQHW products from the marketplace were used to confirm the method. Successful SMRT sequencing results recovered 5416 and 4342 circular-consensus sequencing(CCS) reads belonging to the ITS2 and psb A-trn H regions. The results suggest that with the combination of metabarcoding and SMRT sequencing, it is repeatable, reliable, and sensitive enough to detect species in the THMPs, and the error in SMRT sequencing did not affect the ability to identify multiple prescribed species and several adulterants/contaminants. It has the potential for becoming a valuable tool for the biomonitoring of multi-ingredient THMPs.

A Highly Pathogenic Strain of Porcine Deltacoronavirus Caused Watery Diarrhea in Newborn Piglets

Porcine deltacoronavirus(PDCoV) is a newly identified virus that causes watery diarrhea in newborn piglets and results in significant economic losses to the pig industry. Since first reported in Hong Kong in 2012, PDCoV has been subsequently detected in USA, South Korea, Thailand, and China's Mainland. Here we isolated a strain of PDCoV, named CHN-GD-2016,from the intestinal content of a diseased newborn piglet with severe diarrhea in a pig farm in Guangdong, China. PDCoV CHN-GD-2016 could be identified by immunofluorescence with PDCoV specific rabbit antisera, and typical crown-shaped particles with spiky surface projections of this PDCoV were observed with electron microscopy. Genomic analysis showed that the PDCoV CHN-GD-2016 was closely related to other Chinese PDCoV strains, with the highest sequence similarity with the strain CHN/Tianjin/2016. Importantly, inoculation of newborn piglets with 1×10~5 TCID_(50) of CHN-GD-2016 by oral feeding successfully reproduced clear clinical symptoms, including vomiting, dehydration, and severe diarrhea in piglets. In addition, the virus RNA in rectal swabs from 1 to 7 days post inoculation was detected, macroscopic and microscopic lesions in small intestine were observed, and viral antigen was also detected in the small intestines with immunohistochemical staining. Collectively, the data show in this study confirms that PDCoV is present in Guangdong,China and is highly pathogenic in newborn piglets.

Trends in herbgenomics

From Shen Nong's Herbal Classic(Shennong Bencao Jing) to the Compendium of Materia Medica(Bencao Gangmu) and the first scientific Nobel Prize for the mainland of China, each milestone in the historical process of the development of traditional Chinese medicine(TCM) involves screening, testing and integrating. After thousands of years of inheritance and development,herbgenomics(bencaogenomics) has bridged the gap between TCM and international advanced omics studies, promoting the application of frontier technologies in TCM. It is a discipline that uncovers the genetic information and regulatory networks of herbs to clarify their molecular mechanism in the prevention and treatment of human diseases. The main theoretical system includes genomics, functional genomics, proteomics, transcriptomics, metabolomics, epigenomics, metagenomics, synthetic biology, pharmacogenomics of TCM, and bioinformatics, among other fields. Herbgenomics is mainly applicable to the study of medicinal model plants, genomic-assisted breeding, herbal synthetic biology, protection and utilization of gene resources, TCM quality evaluation and control, and TCM drug development. Such studies will accelerate the application of cutting-edge technologies, revitalize herbal research, and strongly promote the development and modernization of TCM.

Genomic survey of bZIP transcription factor genes related to tanshinone biosynthesis in Salvia miltiorrhiza

Tanshinones are a class of bioactive components in the traditional Chinese medicine Salvia miltiorrhiza, and their biosynthesis and regulation have been widely studied. Current studies show that basic leucine zipper(bZIP) proteins regulate plant secondary metabolism, growth and developmental processes. However, the b ZIP transcription factors involved in tanshinone biosynthesis are unknown.Here, we conducted the first genome-wide survey of the b ZIP gene family and analyzed the phylogeny,gene structure, additional conserved motifs and alternative splicing events in S. miltiorrhiza. A total of 70 Smb ZIP transcription factors were identified and categorized into 11 subgroups based on their phylogenetic relationships with those in Arabidopsis. Moreover, seventeen Smb ZIP genes underwent alternative splicing events. According to the transcriptomic data, the Smb ZIP genes that were highly expressed in the Danshen root and periderm were selected. Based on the prediction of b ZIP binding sites in the promoters and the co-expression analysis and co-induction patterns in response to Ag^+ treatment via quantitative real-time polymerase chain reaction(qRT-PCR), we concluded that Smb ZIP7 and Smb ZIP20 potentially participate in the regulation of tanshinone biosynthesis. These results provide a foundation for further functional characterization of the candidate Smb ZIP genes, which have the potential to increase tanshinone production.

Comparative Genome Analysis of Scutellaria baicalensis and Scutellaria barbata Reveals the Evolution of Active Flavonoid Biosynthesis

Scutellaria baicalensis(S.baicalensis)and Scutellaria barbata(S.barbata)are common medicinal plants of the Lamiaceae family.Both produce specific flavonoid compounds,including baicalein,scutellarein,norwogonin,and wogonin,as well as their glycosides,which exhibit antioxidant and antitumor activities.Here,we report chromosome-level genome assemblies of S.baicalensis and S.barbata with quantitative chromosomal variation(2 n=18 and 2 n=26,respectively).The divergence of S.baicalensis and S.barbata occurred far earlier than previously reported,and a whole-genome duplication(WGD)event was identified.The insertion of long terminal repeat elements after speciation might be responsible for the observed chromosomal expansion and rearrangement.Comparative genome analysis of the congeneric species revealed the species-specific evolution of chrysin and apigenin biosynthetic genes,such as the S.baicalensis-specific tandem duplication of genes encoding phenylalanine ammonia lyase and chalcone synthase,and the S.barbata-specific duplication of genes encoding 4-Co A ligase.In addition,the paralogous duplication,colinearity,and expression diversity of CYP82 D subfamily members revealed the functional divergence of genes encoding flavone hydroxylase between S.baicalensis and S.barbata.Analyzing these Scutellaria genomes reveals the common and species-specific evolution of flavone biosynthetic genes.Thus,these findings would facilitate the development of molecular breeding and studies of biosynthesis and regulation of bioactive compounds.

The Wolfiporia cocos Genome and Transcriptome Shed Light on the Formation of Its Edible and Medicinal Sclerotium

Wolfiporia cocos(F. A. Wolf) has been praised as a food delicacy and medicine for centuries in China. Here, we present the genome and transcriptome of the Chinese strain CGMCC5.78 of W. cocos. High-confidence functional prediction was made for 9277 genes among the 10,908 total predicted gene models in the W. cocos genome. Up to 2838 differentially expressed genes(DEGs)were identified to be related to sclerotial development by comparing the transcriptomes of mycelial and sclerotial tissues. These DEGs are involved in mating processes, differentiation of fruiting body tissues, and metabolic pathways. A number of genes encoding enzymes and regulatory factors related to polysaccharide and triterpenoid production were strikingly regulated. A potential triterpenoid gene cluster including the signature lanosterol synthase(LSS) gene and its modified components were annotated. In addition, five nonribosomal peptide synthase(NRPS)-like gene clusters,eight polyketide synthase(PKS) gene clusters, and 15 terpene gene clusters were discovered in the genome. The differential expression of the velevt family proteins, transcription factors,carbohydrate-active enzymes, and signaling components indicated their essential roles in the regulation of fungal development and secondary metabolism in W. cocos. These genomic and transcriptomic resources will be valuable for further investigations of the molecular mechanisms controlling sclerotial formation and for its improved medicinal applications.

aunalysis of the Genome Sequence of the Medicinal Plant Salvia miltiorrhiza

Dear Editor Salvia miltiorrhiza Bunge （Danshen） is a medicinal plant of the Lamiaceae family, and its dried roots have long been used in traditional Chinese medicine with hydrophilic phenolic acids and tanshinones as pharmaceutically active components （Zhang et al., 2014; Xu et al., 2016）. The first step of tanshinone biosynthesis is bicyclization of the general diterpene precursor （E,E,E）-geranylgeranyl diphosphate （GGPP） to copalyl diphosphate （CPP） by CPP synthases （CPSs）, which is followed by a cyclization or rearrangement reaction catalyzed by kaurene synthase-like enzymes （KSL）. The resulting intermediate is usually an olefin, which requires the insertion of oxygen by cytochrome P450 mono-oxygenases （CYPs） for the final production of diterpenoids （Zi et al., 2014）. While the CPS, KSL, and several early acting CYPs （CYP76AH1, CYP76AH3, and CYP76AK1） for tanshinone biosynthesis have been identified in S. miltiorrhiza （Gao et al., 2009; Guo et al., 2013, 2016; Zi and Peters, 2013）, the majority of the overall biosynthetic pathway, as well as the relevant regulatory factors associated with tanshinone production, remains elusive （Figure 1B）.

Herbgenomics: Decipher molecular genetics of medicinal plants

Medicinal plants synthesize a huge repertoire of specialized compounds as protective metabolites when they are confronted with complex abiotic and biotic conditions.Humans have used medicinal plants to treat ailments and maintain health throughout civilization.Paleontological studies have shown that the application of medicinal plants,such as Ephedra altissima and Centaurea solstitialis,could be dated back to 60,000 years ago,since their fossils were found in the tomb of prehistoric Neanderthals.

Narrower atomic filter at 422.7 nm based on thermal Ca beam

So far, the only Ca magneto-optic atomic filters reported are based on a heated Ca cell, and the transmission spectrum obtained is double-peaked with 3 GHz effective passband limited by Doppler broadening. A magneto-optic atomic filter with single-peaked transmission spectrum of sub-Doppler bandwidth based on the thermal Ca atomic beam is firstly experimentally demonstrated. The fast Ca dipole transition(1S0–1P1) at 422.7 nm, which matches a strong solar Fraunhofer line, was utilized. A folded ninebeam traveling wave configuration is employed, and the maximum transmission efficiency is measured to be 7.3 %.A sub-Doppler transmission bandwidth of 590 MHz, much narrower than the 2.2 GHz Doppler width in the heated atomic cell, is obtained for the first time.

Dyngo-4a通过影响dynamin 2寡聚体的形成来保护老鼠少受轮状病毒感染导致的伤害

轮状病毒是全球范围内导致5岁以下儿童严重腹泻的病原物之一.虽然已通过审批的数个轮状病毒疫苗可以保护50%以上的人群免受轮状病毒侵害,现有的轮状病毒腹泻对症治疗手段主要是补锌和补充水分,还急需针对轮状病毒腹泻的特异性靶向治疗手段以快速缓解病人的痛苦.以大规模药物筛选为手段,我们发现dyngo-4a,一个发动蛋白2(dynamin 2)的抑制剂,可以显著抑制轮状病毒感染导致的细胞病变.而且dyngo-4a还可以抑制轮状病毒在仔鼠肠道内的复制及其复制导致的腹泻.采用小干扰RNA(siRNA)敲除发动蛋白2,而非发动蛋白1,可以显著抑制轮状病毒感染,说明dyngo-4a主要通过影响发动蛋白2的功能来抑制轮状病毒.机制层面,dyngo-4a通过作用于发动蛋白2并引起发动蛋白2寡聚体异常积累来发挥其抗轮状病毒感染的功能.由此,dyngo-4a是潜在的特异性靶向治疗轮状病毒腹泻的候选药物.