Identification of the lysine and histidine transporter family in Camellia sinensis and the characterizations in nitrogen utilization

In plants,the lysine and histidine transporter(LHT)family represent a class of proteins that mediate the uptake,translocation,and utilization of amino acids.The tea plant(Camellia sinensis)is a perennial evergreen with a relatively high level of amino acids.However,systematic identification and molecular characterization of the LHT gene family has rarely been reported in tea plants.In this study,22 CsLHTs were identified from the‘Shuchazao’genome and classified into two groups.The modeled three-dimensional structure and the conserved domains presented a high similarity among the LHTs proteins.Moreover,it was predicted that a few genes were conserved through the analysis of the physiochemical characters,structures and cis-elements in promoters.The expression patterns in tea plants revealed that CsLHT7 was mainly expressed in the roots,and CsLHT4 and CsLHT11 exhibited relatively high expression in both the roots and leaves.Moreover,the expression of all three genes could be induced by organic nitrogen.Additionally,heterogeneous expression of CsLHT4,CsLHT7 and CsLHT11 in Arabidopsis thaliana decreased the aerial parts biomass compared with that in WT plants while significantly increased the rosette biomass only for CsLHT11transgenic plants versus WT plants.Overall,our results provide fundamental information about CsLHTs and potential genes in N utilization for further analysis in tea plants.

Molecular cloning and functional analysis of two calcium associated cuticular protein genes in Neocaridina denticulata sinensis

The body surface of crustaceans is covered with a sturdy shell.The growth and development of crustaceans are realized through molting.Neocaridina denticulata sinensis is a suitable candidate for crustacean scientific research.Two calcium-associated cuticular protein genes,named NdCAP-1 and NdCAP-2,were obtained from N.denticulata sinensis.Molecular docking simulated the binding effect of both proteins and calcium ions.Semi-quantitative reverse transcription PCR results show that NdCAP-1 is expressed in D_(2-4) stage,NdCAP-2 expressed in D_(2-4) and A-B stages,and both were significantly expressed in the cephalothorax cuticle and pereiopods.Then,it was revealed that NdCAP-1 and NdCAP-2 are regulated by NdEcR-mediated 20 E signaling pathways.Knockdown of NdCAP-1 and NdCAP-2 was observed to cause surface defects.The recombinant proteins(rNdCAP-1 and rNdCAP-2),obtained by prokaryotic expression,had calcium-binding and chitin-binding ability,inhibited formation of calcium carbonate precipitate.These results show that calcium-associated cuticular proteins play important roles in cuticle formation and calcification.

Genetic analyses of ancient tea trees provide insights into the breeding history and dissemination of Chinese Assam tea(Camellia sinensis var.assamica)

Chinese Assam tea(Camellia sinensis var.assamica)is an important tea crop with a long history of cultivation in Yunnan,China.Despite its potential value as a genetic resource,its genetic diversity and domestication/breeding history remain unclear.To address this issue,we genotyped 469 ancient tea plant trees representing 26 C.sinensis var.assamica populations,plus two of its wild relatives(six and three populations of C.taliensis and C.crassicolumna,respectively)using 16 nuclear microsatellite loci.Results showed that Chinese Assam tea has a relatively high,but comparatively lower gene diversity(H_(S)=0.638)than the wild relative C.crassicolumna(H_S=0.658).Clustering in STRUCTURE indicated that Chinese Assam tea and its two wild relatives formed distinct genetic groups,with considerable interspecific introgression.The Chinese Assam tea accessions clustered into three gene pools,corresponding well with their geographic distribution.However,New Hybrids analysis indicated that 68.48%of ancient Chinese Assam tea plants from Xishuangbanna were genetic intermediates between the Puer and Lincang gene pools.In addition,10%of the ancient Chinese Assam tea individuals were found to be hybrids between Chinese Assam tea and C.taliensis.Our results suggest that Chinese Assam tea was domesticated separately in three gene pools(Puer,Lincang and Xishuangbanna)in the Mekong River valley and that the hybrids were subsequently selected during the domestication process.Although the domestication history of Chinese Assam tea in southwestern Yunnan remains complex,our results will help to identify valuable genetic resources that may be useful in future tea breeding programs.

中华乌塘鳢(Bostrychus sinensis)白细胞介素8基因的克隆及免疫应答表达分析

由细菌等感染引起的疾病对中华乌塘鳢(Bostrychus sinensis)养殖业的可持续发展造成严重威胁。白细胞介素8(Interleukin-8,IL-8)在调节炎症反应和抗感染免疫方面扮演着重要的作用。利用PCR扩增和克隆测序获得了中华乌塘鳢IL-8(命名为BsIL-8)cDNA序列。BsIL-8 cDNA序列全长1244 bp,包含576 bp的5’端非编码区序列(5’-UTR),312 bp的开放阅读框序列(ORF)和356 bp的3’端非编码区序列(3’-UTR)。BsIL-8共编码103个氨基酸,包含1个由18个氨基酸组成的信号肽和1个由62个氨基酸组成的SCY结构域。其中,SCY结构域包含1个CXC基序(Cys^(30)-Arg^(31)-Cys^(32))以及4个保守的半胱氨酸残基(Cys^(30)、Cys^(32)、Cys^(57)和Cys^(73))。序列分析结果表明,BsIL-8与大弹涂鱼(Boleophthalmus pectinirostris)的IL-8序列相似性和一致性最高(分别为92.4%和86.4%)。基因组织表达分布结果显示,BsIL-8基因在主要组织器官中呈泛在性表达,头肾BsIL-8基础表达量最高,肝脏次之,外周血最低。Poly(I:C)免疫刺激后,BsIL-8在4个重要免疫器官(脾脏、肝脏、头肾和外周血)中的表达量均发生了显著的上调(分别在刺激后3、12、24和24 h时表达量达到最高,P<0.01)。副溶血弧菌(Vibrio parahaemolyticus)感染后,BsIL-8基因在脾脏和外周血中的表达量均显著上调(均在感染后12 h时表达量达到最高,P<0.01),而在肝脏和头肾中的表达量均显著下调(均在感染后12 h时表达量达到最低,P<0.01)。综上所述,BsIL-8基因参与了中华乌塘鳢抗感染免疫应答过程。

Ankylosing spondylitis coexisting with Clonorchis sinensis infection: A case report

BACKGROUND Ankylosing spondylitis(AS)is a chronic immune-mediated inflammatory disease.The prevailing theory links AS onset to infections in susceptible individuals.Furthermore,infections may impair the immune responses.Numerous studies have investigated links between AS and various infections-bacterial,viral,fungal,and other microorganism infections.However,limited attention has been given to the association between AS and Clonorchis sinensis(C.sinensis)infection.CASE SUMMARY A 27-year-old male with a 10-yr history of AS presented to our hospital with inflammatory lower back pain as the primary manifestation.Ten years ago,the patient had achieved a stable condition after treatment with biological agents.However,he experienced a recurrence of lumbosacral pain with an unexplained cause 10 d before hospital admission.A lumbosacral magnetic resonance imaging(MRI)scan revealed bone marrow edema in the left sacroiliac joint,and laboratory indicators were elevated.Moreover,the presence of C.sinensis eggs was detected in the stool.The patient was prescribed praziquantel,resulting in the disap-pearance of C.sinensis eggs in subsequent routine stool tests and relief from lumbosacral pain.A follow-up MRI scan performed after 4 months revealed a reduction in bone marrow edema around the left sacroiliac joint.CONCLUSION C.sinensis infections could potentially trigger the exacerbation of AS.Clinicians should pay attention to investigating the presence of infections.INTRODUCTION Ankylosing spondylitis(AS)is a chronic inflammatory and rheumatic disease resulting from an imbalance between innate and acquired immune responses[1].While it can affect any part of the spine,its primary symptoms are persistent back pain and stiffness in the lower back and pelvis.The prevalence of AS per 10000 individuals is 23.8 in Europe,31.9 in North America,16.7 in Asia,10.2 in Latin America,and 7.4 in Africa[2].Infections commonly occur in the first 3 months and may act as potential triggers for the first symptoms of AS,often manifesting as gastrointestinal,urinary tract,and respiratory infections of microbiological origin[3,4].Clonorchis sinensis(C.sinensis)infection is a severe parasitic disease affecting millions globally,especially prevalent in China,South Korea,the Far East of Russia,and Vietnam,with an estimated 15 million cases[5].Transmission occurs through the consumption of undercooked freshwater fish containing metacercariae.Adult C.sinensis parasites then establish themselves within the human hepatobiliary system[6].C.sinensis infection triggers the activation of sphingosine 1-phosphate receptor 2,leading to the injury and fibrosis of the hepatobiliary[7].Recent research in a rat model found that C.sinensis infection increases the risk of hepatocellular carcinoma by stimulating hepatic progenitor cell proliferation[8].Complications of C.sinensis infection include cholestasis,cholangitis,biliary system fibrosis,and in severe cases,the development of cholangiocarcinoma[9].Consequently,the primary preventive measure is to abstain from consuming raw or undercooked freshwater fish.Praziquantel is the recommended and effective treatment for this infection[10].While there is existing literature on the coexistence of AS and parasitic infections,there is limited research specifically addressing the simultaneous presence of AS and C.sinensis infection.This case report details a rare scenario of AS coexisting with C.sinensis infection,underscoring the potential impact of C.sinensis infection on AS disease activity.

Genome-Wide Identification of ABCC Gene Subfamily Members and Functional Analysis of CsABCC11 in Camellia sinensis

The ATP-binding cassette(ABC)transporter is a gene superfamily in plants.ATP-binding cassette subfamily C(ABCC)protein is a multidrug resistance-associated(MRP)transporter.They play various roles in plant growth,development,and secondary metabolite transport.However,there are few studies on ABCC transporters in tea plants.In this study,genome-wide association study(GWAS)analysis of epigallocatechin gallate(EGCG)content in 108 strains of Kingbird revealed that CsABCCs may be involved in EGCG transport.We identified 25 CsABCC genes at the genomic level of the tea plant,their phylogenetic tree,gene structure,targeted miRNA and other bioinformatics were analyzed.The expression patterns of CsABCCs in eight different tissues and abiotic stress indicate that they have potential roles in regulating the growth,development,and defense of tea plants.The correlation analysis revealed that the expression of the CsABCC11 gene was closely related to the EGCG content in tea buds of 108 strains of the Kingbird,and the subcellular localization experiments in tobacco showed that CsABCC11 protein was localized on the plasma membrane.The virus-induced gene silencing(VIGS)strategy in tea plants further verified that CsABCC11 was involved in EGCG accumulation.Our study laid a foundation for studying the biological function of CsABCC and provided a new candidate molecular marker gene for further EGCG-related variety breeding,which will be of great interest to breeders.

Analysis of NPK in Camellia sinensis, Gliricidia sepium, and Musa acuminata Biomasses for Preparation of an Organic Fertilizers Formula for Young Tea Plants (Musa acuminata) and Studying of Their Nutrient Release Capacity in the Biodegradation Process

This study investigates the nitrogen (N), phosphorous (P), and potassium (K) contents in raw biomasses of Camellia sinensis, Gliricidia sepium, and Musa acuminata. Therein, the highest N and P content was seen in Camellia sinensis 116.80 ± 0.08 mg and 66.00 ± 0.14 mg respectively. The highest K content (106.80 ± 0.04 mg) was observed in Musa acuminata. Next, all three types of plant materials were allowed to decompose in water for 3 weeks, and a sample from each was analyzed for NPK after the 1st, 2nd, and 3rd week during decomposition. A significant increase in the release of N, P, and K by the Camellia sinensis to water (P Musa acuminate were not significantly changed (P > 0.05) over time. The ratio for N:P:K was calculated for raw biomass samples and decomposed samples to find the best fitting N:P:K ratio to apply to young tea plants as organic fertilizers. In addition to that, the microbial insight of these organic compounds was analyzed by observing how microbial population increased with decomposition by the enumeration of the total microbial count. A considerable increment in total microbial count was observed up to 3.28 × 106, 1.21 × 1010, 2.18 × 108, and 6.49 × 107 CFU/ml for Camellia sinensis, Gliricidia sepium, Musa accuminata (leaves), and Musa accuminata (trunk) respectively. The presence of phosphate solubilizing bacteria (PSB) and nitrogen solubilizing bacteria (NSB) throughout the decomposition period was confirmed by their growth on NBRIP and a modified nutrient medium that was specifically designed for the identification of ammonifiers respectively. Prepared fertilizer samples were applied to young tea plants that were grown in the Mawanella area in Sri Lanka (7°15'12.42"N 80°26'47.62"E) and according to the results, it is clear that fertilizer mixture 1 (N:P:K, 10:5:10, tea dust + Gliricidia + banana trunk) and fertilizer mixture 2 (N:P:K, 10:5:10, tea dust + Gliricidia + banana leaves) has the potential to increase the growth of young tea plants.

Mechanism prediction of Astragalus mongholicus Bunge and Angelica sinensis Diels in treating interstitial lung disease based on network pharmacology and molecular docking

Objective:To investigate the mechanism by which Astragalus mongholicus Bunge(AM),and Angelica sinensis Diels(AS)act in interstitial lung disease(ILD)based on computational prediction.Methods:We screened the ingredients of AM and AS in PubMed,the Web of Science,China National Knowledge Infrastructure(CNKI)Databases,etc.Then obtained the potential effective components.By sharing the same molecular with ILD,we got the possible target genes for ILD treatment and constructed components–targets–disease network with Cytoscape software.The CTD(Comparative Toxicogenomics Database)database was used for GO and KEGG enrichment analysis of these target genes.Results:59 active ingredients that can be druggable were chosen from AM,67 active ingredients were chosen from AS.77 overlapping target genes for AM and ILD and 36 overlapping target genes for AS and ILD were acquired.The hub targets of AM were PTGS2,PTGS1,CDK2,MAOA,ESR1,TOP2A,GSK3B,ESR2,PPARG,NOS2,The hub targets of AS were PTGS2,GABRA1,PTGS1,CHRM1,SLC6A2,ADRA1B,ADRAIA,ADRB2,CHRM3,GABRA2,CHRM2.Quercetin,kaempferol,daidzein,pavilion,7-Hydroxycoumarin,and 5-Hydroxycoumarin were the main active ingredients which have more effective targets.Prediction of the protein-protein interaction network showed PTGS2,GSK3B,PPARG,etc.,were the important predicted targets.The enriched KEGG pathways,including the Immune System,Metabolism of lipids and lipoproteins,Cytokine Signaling in the Immune system,Generic Transcription Pathway,The interleukin pathway,Metabolism of proteins,PI3K-Akt signaling pathway,Metabolic pathways,Innate Immune System,Neuroactive ligand-receptor interaction,Metabolism,GPCR downstream signaling,Amine ligand-binding receptors,Class A/1,Calcium signaling pathway.Molecular docking showed that quercetin,kaempferol,daidzein,pavilion,7-Hydroxycoumarin,5-Hydroxycoumarin had good binding activities with PTGS2 and GSK3B,which mainly mediated PI3K/Akt and other important signaling pathways in the pathogenesis of ILD.Conclusion:The components in AS and AM share some common targets,such as PTGS2.AM and AS may ameliorate ILD through the PI3K-Akt signaling pathway which is mediated by GSK3B.PTGS2,PPARG may also be vital target genes in the treatment of ILD with AM and AS.

Geographical differences of leaf traits of the endangered plant Litsea coreana Levl.var.sinensis and its relationship with climate

Seventeen morphological and anatomical characteristics of the leaves were selected from five natural populations to explore the variation in leaf traits of Lits ea core ana var.sinensis and the effects of geographical environment on these variations.Nested analysis of variance,multiple comparisons,principal component analysis(PCA),and correlation analysis were conducted to explore the variations within and between populations and their correlation with geographical and climatic factors.Significant differences in the 17 leaf traits were observed within and among populations.On average,the relative contribution of within population variation to total variation was 24.8%,which was lower than among population variation(54.6%).The average differentiation coefficient of the traits was 65.8%,and the average coefficient of variation 11.8%,ranging from6.7%for main vein thickness to 21.4%for petiole length.The PC A results showed that morphological characteristics were divided into two categories,and the level of variation was greater than that of leaf anatomy.Most of the leaf traits were significantly correlated with geography and climate and showed a gradual variation with longitude,latitude,and altitude.In areas with high temperatures,less rainfall,and strong seasonal rainfall,the leaves are larger,longer and thicker.This study shows that variations in leaf traits of L.coreana var.sinensis mainly come from variations among populations.The level of trait differentiation among populations is high and the level of variation within populations low.These findings help further understand leaf morphological characteristics of this species and can provide a valuable reference for the protection and sustainable utilization of this natural resource.

A likely autotetraploidization event shaped the Chinese mahogany(Toona sinensis)genome

Chinese mahogany(Toona sinensis) is of considerable medical and economic importance, and its genome has been deciphered. However, the process underlying its polyploidy is unclear, and the chromosomal evolutionary trajectory is poorly understood. Here, by reanalysing the T.sinensis genome, we found evidence of a tetraploidization event(T. sinensis special tetraploidization, TST) that occurred approximately 15-17 million years ago(MYA) after the core eudicot-common hexaploidization(ECH or gamma) event. We characterized the synonymous nucleotide substitution rates(Ks values) of collinear genes and found that T. sinensis genes affected by the TST evolve at a slower rate than Acer yangbiense genes. Furthermore, we identified homologous genes related to polyploidization and speciation and constructed multiple alignments with different reference genomes. Notably, the significant balance of gene retention and loss characterized in the two TST-derived subgenomes suggests an autopolyploid nature of the TST. Moreover, we deduced the chromosomal karyotypes of the two subgenomes and identified 7chromosomal fusions that have shaped the T. sinensis genome;more information is available on a newly constructed karyotype platform(http://www.cgrpoee.top/Toona_sinensis/index.html). The T. sinensis genome preserves the ancestral chromosome structure of dicotyledons well and could serve as a good reference for understanding genomic changes in other Meliaceae and Sapindales plants. In addition, we verified that tandem duplication and the ECH have promoted the expansion of terpene synthase(TPS) genes;conversely, the TST seems to have inhibited expansion of these genes. This present effort has clarified the polyploidy events of the T. sinensis genome, filled gaps in the history of karyotype evolution, and laid a solid foundation for further genomic studies in the Meliaceae research community and beyond.

Identifi cation of sesquiterpene synthase genes in the genome of Aquilaria sinensis and characterization of anα-humulene synthase

Sesquiterpenes are the major pharmacodynamic components of agarwood,a precious traditional Chinese medicine obtained from the resinous portions of Aquilaria sinensis trees that form in response to environmental stressors.To characterize the sesquiterpene synthases responsible for sesquiterpene production in A.sinensis,a bioinformatics analysis of the genome of A.sinensis identifi ed six new terpene synthase genes,and 16 sesquiterpene synthase genes were identifi ed as type TPS-a in a phylogenetic analysis.The expression patterns for eight of the sesquiterpene synthase genes after treatment with various hormones or hydrogen peroxide were analyzed by real-time quantitative PCR.The results suggest that 100μM methyl jasmonate,ethephon,(±)-abscisic acid or hydrogen peroxide could be eff ective short-term eff ectors to increase the expression of sesquiterpene synthase genes,while 1 mM methyl salicylate may have long-term eff ects on increasing the expression of specifi c sesquiterpene synthase genes(e.g.,As-SesTPS,AsVS,AsTPS12 and AsTPS29).The expression changes in these genes under various conditions refl ected their specifi c roles during abiotic or biotic stresses.Heterologous expression of a novel A.sinensis sesquiterpene synthase gene,AsTPS2,in Escherichia coli produced a major humulene product,so AsTPS2 is renamed AsHS1.AsHS1 is diff erent from ASS1,AsSesTPS,and AsVS,for mainly producingα-humulene.Based on the predicted space conformation of the AsHS1 model,the small ligand molecule may bind to the free amino acid by hydrogen bonding for the catalytic function of the enzyme,while the substrate farnesyl diphosphate(FPP)probably binds to the free amino acid on one side of the RxR motif.Arg450,Asp453,Asp454,Thr457,and Glu461 from the NSE/DTE motif and D307 and D311 from the DDxxD motif were found to form a polar interaction with two Mg^(2+)clusters by docking.The Mg^(2+)-bound DDxxD and NSE/DTE motifs and the free RXR motif are jointly directed into the catalytic pocket of AsHS1.Comparison of the tertiary structural models of AsHS1 with ASS1 showed that they diff ered in structures in several positions,such as surrounding the secondary catalytic pocket,which may lead to diff erences in catalytic products.Based on the results,biosynthetic pathways for specifi c sesquiterpenes such asα-humulene in A.sinensis are proposed.This study provides novel insights into the functions of the sesquiterpene synthases of A.sinensis and enriches knowledge on agarwood formation.

Differential metabolites and their transcriptional regulation in seven major tea cultivars (Camellia sinensis) in China

Various genetic and biochemical characteristics exist in tea plant cultivars,and they largely determine production suitability and tea quality.Here,we performed transcriptomic and metabolomic analyses of young shoots of seven tea cultivars and identified major regulatory transcription factors(TFs)for the characteristic metabolites in different cultivars based on weighted gene co-expression network analysis(WGCNA).Phenotypically,we found that‘Tieguanyin’(TGY)and‘Fujian Shuixian’(FJSX),which are suitable for oolong tea,had higher catechin contents.The metabolites of‘Jinxuan’(JX)were more prominent,especially the contents of phenolic acids,flavonoids,terpenes,and tannins,which were higher than those of the other six cultivars.Moreover,‘Fudingdabai’(FDDB),which is suitable for white tea,was rich in amino acids,linolenic acid,and saccharides.At the molecular level,hydroxycinnamoyl CoA quinate hydroxycinnamoyl transferase(HCT)(CsTGY12G0001876,and CsTGY06G0003042)led to the accumulation of chlorogenic acid in TGY.The main reason for the higher l-ascorbic acid content in FJSX was the high expression levels of L-galactono-1,4-lactone hydrogenase(GalLDH)(CsTGY13G0000389)and Myo-inositol oxygenase(MIOX)(CsTGY14G0001769,and CsTGY14G0001770),which were regulated by WRKY(CsTGY11G0001197).Furthermore,FDDB,‘Longjing 43’(LJ43),‘Shuchazao’(SCZ)and‘Baihaozao’(BHZ)had higher free fatty acid contents,among which MYB(CsTGY14G0002344)may be a hub gene for the regulation of palmitoleic acid accumulation.More importantly,we found that the shoots of TGY were green with purple,mainly due to the accumulation of anthocyanins and the downregulation of the Mg-protoporphyrin IX nonomethyl ester cyclase(MPEC)(CsTGY10G0001989)gene that affects chlorophyll synthesis.These results will provide a theoretical reference for tea cultivar breeding and suitability.

Identification of suitable reference genes for quantitative gene expression analysis in clam Cyclina sinensis under salinity stress and Vibrio infection

The appropriate reference gene is a prerequisite for accurate normalization of gene expression level,and research on suitable reference genes in clam Cyclina sinensis is scarce.To improve the situation,we selected five commonly used housekeeping genes,including β-actin,Elongation factor 1-α(EF1-α),Glyceraldehyde-3-pho sphate dehydrogenase(GAPDH),40S ribosomal protein S18(RPS18),and Tubulin a(TUB-α),then evaluated their expression stability in different adult tissues and under different experimental treatments(salinity stress and Vibrio parahaemolyticus infection).Their expression stability was analyzed by three frequently used programs,geNorm,NormFinder,and BestKeeper.This analysis indicated that multiple genes should be used for normalization,and we concluded that the reference gene combination GAPDH-RPS18-β-actin,should be used for qRT-PCR analysis in different tissues of C.sinensis under normal physiological conditions.For the clams under salinity stress and Vibrio infection,EF1-α-GAPDHRPS18 was recommended as the gene combination for qRT-PCR normalization.TUB-αwas generally poorly ranked by all programs,and should not be used in future studies.This study should provide fundamental support for accurate quantitative gene expression analysis of this species.

Natural variation of main biochemical components,morphological and yield traits among a panel of 87 tea [Camellia sinensis(L.) O. Kuntze] cultivars

Many attentions have been previously focused to identify the multiple biochemical components related to tea quality and health benefits,however, the natural variation of biochemical components present in tea germplasm has not been adequately evaluated. In this study, the main biochemical components, leaf morphological and yield characteristics were evaluated for four rounds of tea leaves in a panel of 87 elite tea cultivars suitable for black, green, or oolong tea. Significant variations were observed among the tea cultivars, as well as seasonal differences in the levels of the free amino acid(FAA), caffeine(CAF), tea polyphenols(TP), water extract(WE) and TP to FAA ratio(TP/FAA). Results showed that the average levels of FAA showed a seasonal change, with the highest level of 4.0% in the 1st spring tea in the cultivars suitable for green tea and the lowest of 3.2% in summer tea in the cultivars suitable for black tea. The average CAF content was highest 3.2% in the cultivars suitable for oolong tea in the 1st spring and the lowest 2.5% in the cultivars suitable for green tea in summer. Limited seasonal and varietal variations were noticed in the average levels of WE among the three categories of tea. In addition, significant natural variation of the morphological characteristics, bud length varying from 2.5 cm to 8.7 cm, bud density from 190.3 buds · m-2to 1 730.3 buds · m-2, mature leaves biomass from 128.4 kg · hm-2to 2 888.4 kg · hm-2, and yield component traits of 100 buds(one bud with two leaves) dry weight from 3.7 g to37.7 g, tea yield/round from 444.6 kg · hm-2to 905.3 kg · hm-2, were observed. The aim of our evaluation was not only to identify the advantages of seasonal and clonal variations but also to provide a new viewpoint for their further application. Representative accessions were selected from the germplasm to promote the establishment of an inherent biochemical constituent expressing the quality of black, green, and oolong tea. The findings might be utilized to establish early selection criteria to enhance the tea breeding and production program.

Population dynamics and protection strategies of Litsea coreana Levl.var.sinensis with Guizhou Province in China as an example

Studying the dynamics of plant populations is crucial to the conservation and management of endangered plants.The diameter at breast height(DBH),tree height,crown width,and environmental factors of a total of 15 plots in five populations in Guizhou Province,China were measured in this study.The method of substituting diameter at breast height for age was used to divide the population into 7 age classes.The age structure,static life table,and relationship between the population characteristics and environmental factors of the Litsea coreana Levl.var.sinensis population were studied.Moreover,the time series model was used to predict the population dynamics.Results showed that the age structure of population in Zhengan County(Population ZA)had an inverted Jshape with abundant seedlings and insufficient middle-aged and elderly individuals.The age structures of populations in Daozhen County(Population DZ),Kaiyang County(KY),Meitan County(Population MT),and Xishui County(Population XS)were spindle-shaped with few young individuals and many middle-aged and elderly individuals.The mortality rate of each population fluctuated with the increase in age class,and the populations were extremely unstable.The survival curve was close to the Deevey III type,and the degree of deviation was related to the lack of young individuals.After the age stages II,IV,and VI,the number of aged plants in the different populations initially increased then decreased,and population stability was difficult to maintain.Redundancy analysis showed that the DBH,tree height and crown width of the population decreased with the increase of annual mean temperature,and the population density was restricted by soil nutrient conditions to a certain extent.The results of this work provide scientific basis for the conservation and sustainable utilization of natural populations of L.coreana var.sinensis in Guizhou Province,China.

The high-resolution three-dimensional(3D)chromatin map of the tea plant(Camellia sinensis)

Dear Editor,Tea plant(Camellia sinensis(L.)O.Kuntze)is one of the world’s most important non-alcoholic beverages,with great economic,health,and cultural value.Recently,several genomes of C.sinensis var.assamica(CSA)(Yunkang 10),C.sinensis var.sinensis(CSS)(Shuchazao,Biyun,Longjing 43,Tieguanyin,Huangdan),and ancient tea plant(DASZ)have been deciphered[1-3].

Monitoring Thosea sinensis Walker in Tea Plantations Based on UAV Multi- Spectral Image

Thosea sinensis Walker(TSW)rapidly spreads and severely damages the tea plants.Therefore,finding a reliable operational method for identifying the TSW-damaged areas via remote sensing has been a focus of a research community.Such methods also enable us to calculate the precise application of pesticides and prevent the subsequent spread of the pests.In this work,based on the unmanned aerial vehicle(UAV)platform,five band images of multispectral red-edge camera were obtained and used for monitoring the TSW in tea plantations.By combining the minimum redundancy maximum relevance(mRMR)with the selected spectral features,a comprehensive spectral selection strategy was proposed.Then,based on the selected spectral features,three classic machine learning algorithms,including random forest(RF),support vector machine(SVM),and k-nearest neighbors(KNN)were used to construct the pest monitoring model and were evaluated and compared.The results showed that the strategy proposed in this work obtained ideal monitoring accuracy by only using the combination of a few optimized features(2 or 4).In order to differentiate the healthy and TSW-damaged areas(2-class model),the monitoring accuracies of all the three models were computed,which were above 96%.The RF model used the least number of features,including only SAVI and Bandred.In order to further discriminate the pest incidence levels(3-class model),the monitoring accuracies of all the three models were computed,which were above 80%,among which the RF algorithm based on SAVI,Band_(red),VARI__(green),and Band_(red_edge) features achieve the highest accuracy(OAA of 87%,and Kappa of 0.79).Considering the computational cost and model accuracy,this work recommends the RF model based on a few optimal feature combinations to monitor and distinguish the severity of TSW in tea plantations.According to the UAV remote sensing mapping results,the TSW infestation exhibited an aggregated distribution pattern.The spatial information of occurrence and severity can offer effective guidance for precise control of the pest.In addition,the relevant methods provide a reference for monitoring other leaf-eating pests,effectively improving the management level of plant protection in tea plantations,and guaranting the yield and quality of tea plantations.

Quality Standard of Oxyria sinensis Hemsl

[Objectives]To establish the quality standard of Oxyria sinensis Hemsl.[Methods]The method in Chinese Pharmacopoeia(2020 edition)was used to determine the moisture,total ash,acid-insoluble ash and alcohol-soluble extracts of O.sinensis.The qualitative identification of medicinal materials was carried out by microscopic identification method and thin layer chromatography(TLC).The contents of hyperoside and isoquercitrin were determined by high performance liquid chromatography(HPLC).[Results]The total amount of moisture,total ash,acid-insoluble ash,alcohol-soluble extract,hyperoside and isoquercitrin of 12 batches of medicinal materials were 8.56%-11.53%,11.26%-15.48%,1.62%-2.56%,17.75%-26.09%,and 0.036%-0.42%,respectively.The microscopic features were obvious,and pores,fibers,calcium oxalate clusters,calcium oxalate square crystals,cork cells could be observed.Fluorescent spots of the same color were displayed at the positions corresponding to the hyperoside reference substance.[Conclusions]Establishing relatively complete quality standards for O.sinensis medicinal materials can provide a reference for the quality control of medicinal materials.

Research progress on the application of the main components of Cuscuta sinensis in the urogenital system

Cuscuta sinensis is a traditional Chinese medicine commonly used in traditional medicine,known for its liver and kidney tonifying effects,sperm fixing and urine reduction properties,fetus calming abilities,eye brightening qualities,and diarrhea stopping capabilities.Modern pharmacological studies have focused on its chemical components such as flavonoids,polysaccharides,alkaloids,steroids,and other bioactive compounds.These studies have revealed its various pharmacological effects,including hormone secretion regulation,ovulation promotion,sperm protection,testicular development promotion,and sperm formation.It is extensively utilized in the treatment of urological and reproductive system diseases,such as male sexual dysfunction,polycystic ovary syndrome,premature ovarian failure,weak spermatozoa,and infertility.This article provides a review of recent advancements in the application of Cuscuta sinensis in the genitourinary system,aiming to enhance the clinical application of Cuscuta sinensis preparations.

Mechanism of Cordyceps sinensis in atherosclerosis treatment based on network pharmacology and molecular docking analysis

Background:The present study intented to delve into the molecular mechanism of Cordyceps sinensis(C.sinensis)in treating atherosclerosis by combining network pharmacology and molecular docking analysis.Methods:We searched the databases including Traditional Chinese Medicines Systems Pharmacology Database and Analysis Platform,PubChem,and PharmMapper to screen out the active chemical ingredients of C.sinensis and the corresponding targets.The String database was used for the matching normalization of results,and the software Cytoscape 3.7.2 was adopted to establish the C.sinensis-active components-targets of action-disease network.The databases of Online Mendelian Inheritance in Man database,GeneCards,Therapeutic Target Database,and DisGNET were searched to yield the major targets of atherosclerosis(AS),which were matched with the active component targets of C.sinensis to identify the potential therapeutic targets.The String database was utilized to set up the protein-protein interaction network,and Cytoscape software was applied for topological analysis,which was followed by the Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes signaling pathway analysis based on the DAVID database.Finally,the core components of C.sinensis and the targets of action were confirmed via molecular docking on AutoDock Vina and PyMOL.Results:In total,7 bioactive ingredients of C.sinensis were identified from Traditional Chinese Medicines Systems Pharmacology Database and Analysis Platform database and 319 predicted targets were obtained,231 of which were associated with AS.The core targets involved in AS treatment with C.sinensis included MAPK1,SRC,PIK3R1,AKT1,and HSP90AA1.The enrichment analysis unveiled the primary pathways involved in these processes,such as pathways in cancer and lipid and atherosclerosis.Moreover,through molecular docking,it was found that the active ingredients of C.sinensis presented with strong binding capacities with their corresponding targets,and the strongest binding capacity was observed between peroxyergosterol and SRC.Conclusions:The present study revealed at the molecular level that C.sinensis played its role in AS treatment through multiple drug active components,targets of action and pathways,which would point out the direction and provide theoretic basis for future research.