Five New Oleanane Triterpene Saponins from Camellia petelotii and Their Alpha-glucosidase Inhibitory Activity.

Five new triterpenoid glycosides, named campetelosides A-E (1-5), together with three known compounds, chikusetsusaponin IVa (6), umbellatoside B (7), and silvioside E (8) were isolated from the leaves of Camellia petelotii (Merr.) Sealy. Their chemical structures were determined by interpretations of HR-ESI-MS and NMR spectra. In addition, compounds 1-8 were evaluated for their α-glucosidase inhibitory effects. Compounds 1-3 significantly showed α-glucosidase inhibitory activity with IC50 values of 166.7±6.0, 45.9±2.6, and 395.3±10.5 µM, respectively, compared to that of the positive control, acarbose, with an IC50 value of 200.4±10.5 µM.

A new oleanane-type triterpenoid saponin with α-glucosidase inhibitory activity from Camellia nitidissima.

A new oleanane-type triterpenoid saponin, 21ß, 22α-di-O-angeloyl-15α, 16α, 28-trihydroxyolean-12-ene 3ß-O-α-L-rhamnopyranosyl-(1→3)-α-D-xylopyranosyl-(1→3)-ß-D-glucopyranoside (1), together with five known compounds (2-5), were isolated from Camellia nitidissima. Their structures were elucidated based on spectroscopic methods, including extensive NMR and MS spectra. Compound 1 showed potential inhibitory activity on α-glucosidase with the IC50 values of 185.9 ± 44.5 µmol/L.

Phylotranscriptomics of Theaceae: generic-level relationships, reticulation and whole-genome duplication.

BACKGROUND AND AIMS: Theaceae, with three tribes, nine genera and more than 200 species, are of great economic and ecological importance. Recent phylogenetic analyses based on plastomic data resolved the relationships among the three tribes and the intergeneric relationships within two of those tribes. However, generic-level relationships within the largest tribe, Theeae, were not fully resolved. The role of putative whole-genome duplication (WGD) events in the family and possible hybridization events among genera within Theeae also remain to be tested further. METHODS: Transcriptomes or low-depth whole-genome sequencing of 57 species of Theaceae, as well as additional plastome sequence data, were generated. Using a dataset of low-copy nuclear genes, we reconstructed phylogenetic relationships using concatenated, species tree and phylogenetic network approaches. We further conducted molecular dating analyses and inferred possible WGD events by examining the distribution of the number of synonymous substitutions per synonymous site (Ks) for paralogues in each species. For plastid protein-coding sequences , phylogenies were reconstructed for comparison with the results obtained from analysis of the nuclear dataset. RESULTS: Based on the 610 low-copy nuclear genes (858 606 bp in length) investigated, Stewartieae was resolved as sister to the other two tribes. Within Theeae, the Apterosperma-Laplacea clade grouped with Pyrenaria, leaving Camellia and Polyspora as sister. The estimated ages within Theaceae were largely consistent with previous studies based mainly on plastome data. Two reticulation events within Camellia and one between the common ancestor of Gordonia and Schima were found. All members of the tea family shared two WGD events, an older At-γ and a recent Ad-ß; both events were also shared with the outgroups (Diapensiaceae, Pentaphylacaceae, Styracaceae and Symplocaceae). CONCLUSIONS: Our analyses using low-copy nuclear genes improved understanding of phylogenetic relationships at the tribal and generic levels previously proposed based on plastome data, but the phylogenetic position of the Apterosperma-Laplacea clade needs more attention. There is no evidence for extensive intergeneric hybridization within Theeae or for a Theaceae-specific WGD event. Land bridges (e.g. the Bering land bridge) during the Late Oligocene may have permitted the intercontinental plant movements that facilitated the putative ancient introgression between the common ancestor of Gordonia and Schima.

The specific DNA barcodes based on chloroplast genes for species identification of Theaceae plants.

More than 600 species in over 40 genera have been identified in family Theaceae worldwide. The accurate identification of Theaceae plants can ensure the market economic order, and it plays a vital role in achieving the sustainable utilization of germplasm resources. DNA barcoding, one of the most potential species identification technologies at present, has advanced in the rapid, accurate and repetitive discrimination of species. In this study, matK + ndhF + ycf1 was observed as the optimal combined candidate gene sequence of DNA barcodes by analyzing genetic information of four single chloroplast DNA sequences, including matK, rbcL, ndhF and ycf1, as well as six combined gene sequences. Subsequently, the experiments were performed on phylogenetic analysis based on genetic distance to study the phylogenetic relationship of Theaceae plants and evaluate the species identification accuracy of matK + ndhF + ycf1. Lastly, the species-specific DNA barcodes were designed by searching the variable sites (one type of single nucleotide polymorphism sites) for the accurate identification of Camellia amplexicaulis, Franklinia alatamaha, Gordonia brandegeei and Stewartia micrantha. The previous methods of screening and testing candidate gene sequences were optimized, and innovation was made in the above methods. The process of making visual DNA barcodes was standardized. Besides, DNA barcoding technology increased the accuracy of species identification and DNA barcoding was analyzed in accordance with the theories of population genetics (e.g., neutral theory of molecular evolution). The results of the study will lay a basis for the identification and protection of Theaceae species and germplasm resources. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01175-7.

Analytical Assessment of Bioelements in Various Types of Black Teas from Different Geographical Origins in View of Chemometric Approach.

A comprehensive approach to the mineral composition of black teas of different origins was studied using the Flame Atomic Absorption Spectrometry (FAAS) method, supported by chemometric tools including Principal Component Analysis PCA) and Classification and Regression Trees (CART). Significant differences between the teas from seven countries (Japan, Nepal, Kenya, Iran, Sri Lanka, India, and China) were shown. K was the main element determined in all teas, with an average concentration of 11,649 mg/kg, followed by Ca, Mg and Mn. In general, regarding all investigated black teas, the element content was ranked in the following order: K > Ca > Mg > Mn > Fe > Na > Zn > Cu. The applied chemometric methods allowed us to recognize black tea clusters based on their mineral composition and place of cultivation, and allowed us to find correlations between particular elements in black teas. The performed analyses revealed interesting correlations between the concentration of various elements in black teas: K was negatively correlated with Na, Fe, Mn and Cu; K was positively correlated with the content of Ca and Mg. Significant positive correlations between Mn and Fe and Mn and Zn in the studied black tea samples were also revealed. It was shown that mineral composition may be a significant factor regarding the origin of the black tea, not only considering the country, but also the region or province.

The Role of Extracting Solvents in the Recovery of Polyphenols from Green Tea and Its Antiradical Activity Supported by Principal Component Analysis.

Green tea contains a variety of biologically active constituents that are widely used in the pharmaceutical and food industries. Among them, simple catechins constitute a major group of compounds that is primarily responsible for the high biologic activity of green tea extracts. Therefore, the application of optimized extraction conditions may result in obtaining high value extracts. The main purpose of the study was to compare the content of polyphenols, mainly catechins, and the antioxidant activity of green tea extracts obtained by three different extraction methods: simple maceration, ultrasound extraction and accelerated solvent extraction using six various solvent systems. The quality of the extracts was evaluated by LC-ESI-Q-TOF-MS methodologies and spectrophotometric determinations. The obtained results revealed that catechins' extraction efficiency was identical for the three techniques studied. However, larger quantitative differences among the samples were observed when using different solvents. The total content of major catechins and gallic acid was within a very wide range of 10.2-842 mg/L. Ethyl acetate was by far the least effective extractant, regardless of the extraction technique used. After all, the solvent system composed of ethanol:water (1:1 v/v) was proven to be the best to recover catechins and to deliver extracts with the highest antiradical activity.

A high-density genetic map of Schima superba based on its chromosomal characteristics.

BACKGROUND: Schima superba (Theaceae) is a popular woody tree in China. The obscure chromosomal characters of this species are a limitation in the development of high-density genetic linkage maps, which are valuable resources for molecular breeding and functional genomics. RESULTS: We determined the chromosome number and the karyotype of S. superba as 2n = 36 = 36 m, which is consistent with the tribe Schimeae (n = 18). A high-density genetic map was constructed using genotyping by sequencing (GBS). A F1 full-sib with 116 individuals and their parents (LC31 × JO32) were sequenced on the Illumina HiSeq™ platform. Overall, 343.3 Gb of raw data containing 1,191,933,474 paired-end reads were generated. Based on this, 99,966 polymorphic SNP markers were developed from the parents, and 2209 markers were mapped onto the integrated genetic linkage map after data filtering and SNP genotyping. The map spanned 2076.24 cM and was distributed among 18 linkage groups. The average marker interval was 0.94 cM. A total of 168 quantitative trait loci (QTLs) for 14 growth traits were identified. CONCLUSIONS: The chromosome number and karyotype of S. superba was 2n = 36 = 36 m and a linkage map with 2209 SNP markers was constructed to identify QTLs for growth traits. Our study provides a basis for molecular-assisted breeding and genomic studies, which will contribute towards the future research and genetic improvement of S. superba.

Applications of Tea (Camellia sinensis) and its Active Constituents in Cosmetics.

Studies on the cosmetic applications of plant extracts are increasingly appearing in the scientific literature, which is due to the growing popularity of skincare products around the world. In the light of the observed changes, a return to natural treatment and skincare with cosmetics free of harmful substances or toxic preservatives is visible. Currently, tea extracts, due to their rich composition and various biological actions, play an important role among the dietary supplements and cosmetics. This review is intended to collect the reports on the properties of the tea plant, its extracts and preparations in cosmetology: for skin care products and for the treatment of selected dermatological diseases. Particular attention is paid to its antioxidant, anti-hyaluronidase, anti-inflammatory, slimming, hair-strengthening, photoprotective and sealing blood vessels properties.

Phylogeny, historical biogeography, and diversification of angiosperm order Ericales suggest ancient Neotropical and East Asian connections.

Inferring interfamilial relationships within the eudicot order Ericales has remained one of the more recalcitrant problems in angiosperm phylogenetics, likely due to a rapid, ancient radiation. As a result, no comprehensive time-calibrated tree or biogeographical analysis of the order has been published. Here, we elucidate phylogenetic relationships within the order and then conduct time-dependent biogeographical and diversification analyses by using a taxon and locus-rich supermatrix approach on one-third of the extant species diversity calibrated with 23 macrofossils and two secondary calibration points. Our results corroborate previous studies and also suggest several new but poorly supported relationships. Newly suggested relationships are: (1) holoparasitic Mitrastemonaceae is sister to Lecythidaceae, (2) the clade formed by Mitrastemonaceae + Lecythidaceae is sister to Ericales excluding balsaminoids, (3) Theaceae is sister to the styracoids + sarracenioids + ericoids, and (4) subfamilial relationships with Ericaceae suggest that Arbutoideae is sister to Monotropoideae and Pyroloideae is sister to all subfamilies excluding Arbutoideae, Enkianthoideae, and Monotropoideae. Our results indicate Ericales began to diversify 110 Mya, within Indo-Malaysia and the Neotropics, with exchange between the two areas and expansion out of Indo-Malaysia becoming an important area in shaping the extant diversity of many families. Rapid cladogenesis occurred along the backbone of the order between 104 and 106 Mya. Jump dispersal is important within the order in the last 30 My, but vicariance is the most important cladogenetic driver of disjunctions at deeper levels of the phylogeny. We detect between 69 and 81 shifts in speciation rate throughout the order, the vast majority of which occurred within the last 30 My. We propose that range shifting may be responsible for older shifts in speciation rate, but more recent shifts may be better explained by morphological innovation.

Two new ent-kaurane diterpenes from the stems of Eurya chinensis.

Two new ent-kaurane diterpenes (1-2), together with five known analogs, were isolated from the stems of Eurya chinensis. The structures of new compounds were established by extensive analysis of mass spectrometric and 1D and 2D NMR spectroscopic data. Compound 3 exhibited noticeable anti-inflammatory activity as denoted by inhibiting LPS-induced nitric oxide (NO) production in RAW264.7 cells with an IC50 value of 7.82 µM. Compound 4 showed potent cytotoxic activity against human cancer cell lines NCI-H46, HepG2 and SW480 with IC50 values ranging from 7.45 to 8.54 µM.

Insights into the historical assembly of East Asian subtropical evergreen broadleaved forests revealed by the temporal history of the tea family.

Subtropical evergreen broadleaved forests (EBLFs) inhabit large areas of East Asia. Although paleovegetation reconstructions have revealed that the subtropical EBLFs existed in Southwest China during the Miocene, the historical construction of these forests remains poorly known. Here, we used the tea family (Theaceae), a characteristic component of the subtropical EBLFs, to gain new insights into the assembly of this important biome. Using a robust phylogenetic framework of Theaceae based on plastome and nuclear ribosomal DNA sequence data, the temporal history of the family was reconstructed. Data from other characteristic components of subtropical EBLFs, including Fagaceae, Lauraceae and Magnoliaceae, were also integrated. Most of the essential elements of the subtropical EBLFs appear to have originated around the Oligocene-Miocene (O-M) boundary. However, small woody lineages (e.g. Camellia, Hartia) from Theaceae were dated to the late Miocene. Accelerated net diversification rates within Theaceae were also detected near the O-M transition period and the late Miocene. Our results suggest that two independent intensifications of the East Asian summer monsoon (EASM) around the O-M boundary and the late Miocene may have facilitated the historical assembly of the subtropical EBLFs in East Asia.

Nectar properties and the role of sunbirds as pollinators of the golden-flowered tea (Camellia petelotii).

PREMISE OF THE STUDY: Properties of floral nectar have been used to predict if a plant species is pollinated by birds. To see whether winter-flowering plants evolve nectar properties corresponding to bird pollinators, nectar properties of several Camellia species (including the golden-flowered tea), as well as the role of floral visitors as effective pollinators, were examined. METHODS: Potential pollinators of Camellia petelotii were identified at different times of day and under various weather conditions. A bird exclusion experiment was used to compare the pollination effectiveness of birds and insects. Nectar sugar components (fructose, glucose, and sucrose) from C. petelotii growing wild and another seven Camellia species and 22 additional cultivars (all in cultivation) were examined by high-performance liquid chromatography (HPLC). KEY RESULTS: The sunbird Aethopyga siparaja and honeybees were the most frequent floral visitors to C. petelotii. Honeybee visits were significantly reduced in cloudy/rainy weather. The fruit and seed set of flowers with birds excluded were reduced by 64%, indicating that bird pollination is significant. For the wild populations of C. petelotii, a bagged flower could secrete 157 µL nectar; this nectar has a low sugar concentration (19%) and is sucrose-dominant (87%). The eight Camellia species and 22 cultivars had an average sugar concentration of around 30% and a sucrose concentration of 80%, demonstrating sucrose-dominant nectar in Camellia species. CONCLUSIONS: The nectar sugar composition of Camellia species was characterized by sucrose dominance. In addition, the large reduction in seed set when birds are excluded in the golden-flowered tea also supports the suggestion that these winter-flowering plants may have evolved with birds as significant pollinators.

Triterpene saponins and megastigmane glucosides from Camellia bugiamapensis.

Two new triterpene saponins, camelliosides I and J (1 and 2), two new megastigmane glycosides, camellistigosides A and B (3 and 4), and two known megastigmane glycosides, icariside B1 (5) and (6S,9R)-roseoside (6), were isolated from a methanol extract of the Camellia bugiamapensis leaves using various chromatographic separation techniques. Their structures were elucidated based on spectroscopic analyses, including HR ESI MS, CD, 1D and 2D NMR. Their inhibitory effects on LPS-induced NO production in RAW264.7 cells were evaluated. This is the first report of the chemical constituents and biological activity of C. bugiamapensis.

Anti-inflammatory and antimicrobial coumarins from the stems of Eurya chinensis.

Two new coumarins, named (±)-euryacoumarin A (1) and 6-demethylobtusinin (2), and one new natural coumarin, named euryacoumarin B (3), along with two known compounds, scopoletin (4) and obtusinol (5), were isolated from the stems of Eurya chinensis. Their structures were elucidated by means of extensive spectroscopic methods and comparison with data reported in the literatures. Compound 1 exhibited significant inhibition of LPS-induced nitric oxide (NO) production in RAW264.7 cells with IC50 value of 35.64 ± 1.73 µM, and showed marginal antibacterial activities against Bacillus subtilis and B. cereus with MIC values of 50.59 ± 2.12 and 35.42 ± 0.96 µM, respectively.

Transcriptome analysis reveals self-incompatibility in the tea plant (Camellia sinensis) might be under gametophytic control.

BACKGROUND: Self-incompatibility (SI) is under genetic control and prevents inbreeding depression in angiosperms. SI mechanisms are quite complicated and still poorly understood in many plants. Tea (Camellia sinensis L.) belonging to the family of Theaceae, exhibits high levels of SI and high heterozygosity. Uncovering the molecular basis of SI of the tea plant may enhance breeding and simplify genomics research for the whole family. RESULTS: The growth of pollen tubes following selfing and crossing was observed using fluorescence microscopy. Self-pollen tubes grew slower than cross treatments from 24 h to 72 h after pollination. RNA-seq was employed to explore the molecular mechanisms of SI and to identify SI-related genes in C. sinensis. Self and cross-pollinated styles were collected at 24 h, 48 h and 72 h after pollination. Six RNA-seq libraries (SP24, SP48, SP72, CP24 CP48 and CP72; SP = self-pollinated, CP = cross-pollinated) were constructed and separately sequenced. In total, 299.327 million raw reads were generated. Following assembly, 63,762 unigenes were identified, and 27,264 (42.76 %) unigenes were annotated in five public databases: NR, KOG, KEGG, Swiss-Port and GO. To identify SI-related genes, the fragments per kb per million mapped reads (FPKM) values of each unigene were evaluated. Comparisons of CP24 vs. SP24, CP48 vs. SP48 and CP72 vs. SP72 revealed differential expression of 3,182, 3,575 and 3,709 genes, respectively. Consequently, several ubiquitin-mediated proteolysis, Ca(2+) signaling, apoptosis and defense-associated genes were obtained. The temporal expression pattern of genes following CP and SP was analyzed; 6 peroxidase, 1 polyphenol oxidase and 7 salicylic acid biosynthetic process-related genes were identified. The RNA-seq data were validated by qRT-PCR of 15 unigenes. Finally, a unigene (CL25983Contig1) with strong homology to the S-RNase was analyzed. It was mainly expressed in styles, with dramatically higher expression in self-pollinated versus cross-pollinated tissues at 24 h post-pollination. CONCLUSIONS: The present study reports the transcriptome of styles after cross- and self-pollination in tea and offers novel insights into the molecular mechanism behind SI in C. sinensis. We believe that this RNA-seq dataset will be useful for improvement in C. sinensis as well as other plants in the Theaceae family.

Camellia nanningensis sp. nov.: the earliest fossil wood record of the genus Camellia (Theaceae) from East Asia.

A new species Camellia nanningensis was described on the basis of well-preserved mummified wood from the upper Oligocene Yongning Formation of Nanning Basin in Guangxi Province, South China. This represents the most ancient fossil wood assigned to Camellia, and the earliest fossil record of the family Theaceae in China. This fossil material shows that Camellia occurred in China as early as the late Oligocene, suggesting more ancient radiation of this genus than estimated by molecular dating.

Two new 28-nor-oleanane-type triterpene saponins from roots of Camellia oleifera and their cytotoxic activity.

Two new 28-nor-oleanane-type triterpene saponins, oleiferoside U (1), and oleiferoside V (2) were isolated from the 50% EtOH extract of the roots of Camellia oleifera C. Abel. Their structures were elucidated as camellenodiol 3ß-O-ß-d-galactopyranosyl-(1→2)-ß-d-xylopyranosyl-(1→2)-[ß-d-galactopyranosyl-(1→3)]-ß-d-glucuronopyranoside and camellenodiol 3ß-O-ß-d-galactopyranosyl-(1→3)-ß-d-xylopyranosyl-(1→2)-[ß-d-galactopyranosyl-(1→3)]-ß-d-glucuronopyranoside. Their chemical structures were established mainly on the basis of integrated spectroscopic techniques. In vitro, cytotoxic activities of the two new triterpene saponins were evaluated against three human tumor cell lines (A549, SMMC-7721, and MCF-7) using the MTT assay. Both of them showed a certain cytotoxic activities toward the tested cell lines and gave IC50 values in the range of 45.04-63.22 µM.

The occurrence and distribution of Tuckerella japonica (Acari: Tuckerellidae) on tea bushes, Camellia sinensis and C. assamica, in Alabama, Georgia and South Carolina, USA.

Adults, immatures and eggs of Tuckerella japonica (Ehara) were collected from unknown clones or varieties of Camellia sinensis (L.) O. Kuntze tea bushes in the Clemson University Farm, Coastal Research and Education Center, Charleston, South Carolina; from Assam hybrids in The Caw Caw Nature Preserve in Ravenel, SC; from C. sinensis and C. assamica (Masters) in the Charleston Tea Plantation on Wadmalaw Island, SC; C. sinensis in the Fairhope Tea Plantation in Fairhope, Alabama; and from C. sinensis 'Rosea' and a C. sinensis and C. assamica hybrid in Savannah and Ellabell, Georgia, between 1994 and 2015. This mite was consistently collected from 1-, 2- and 3+-year-old wood of tea plants with significantly greater numbers collected from 2-year-old wood. All stages of the mite were found within longitudinally split areas of the wood where underlying green bark tissues were exposed. As 1-year-old wood matured, there was increased splitting of the bark with increased mite presence. Mature green fruit (= developing seed pods) of tea were also frequented by T. japonica between June-July and October and their numbers were no greater than those on 1- or 3+-year wood. When the fruit were small (March-May) or as they hardened in late fall, they were not suitable feeding sites for this mite. Very few T. japonica were collected from 50 mature, inner or outer leaf samples with none usually found. Tuckerella japonica has multiple, overlapping generations and occurs on tea throughout the year in Alabama, Georgia and South Carolina, USA.

Two triterpenoid glycosides from the roots of Camellia oleifera and their cytotoxic activity.

Two new triterpenoid saponins, oleiferoside N and oleiferoside O, were isolated from the EtOH extract from the roots of Camellia oleifera C. Abel. Their structures were elucidated as 16α-acetoxy-21ß,22α-O-diangeloyloxy-23,28-dihydroxyolean-12-ene 3ß-O-ß-D-xylopyranosyl-(1 → 3)-α-L-arabinopyranosyl-(1 → 3)-ß-D-glucuronopyranoside (1), 16α-acetoxy-21ß-O-angeloyloxy-23,28-dihydroxy-22α-O-(2-methylbutanoyloxy)olean-12-ene 3ß-O-ß-D-xylopyranosyl-(1 → 3)-α-L-arabinopyranosyl-(1 → 3)-ß-D-glucuronopyranoside (2), on the basis of spectroscopic analyses (IR, ESI-MS, HR-ESI-MS, 1D and 2D NMR) and acid hydrolysis. Both were characterized to be oleanane-type saponins with sugar moieties linked to C-3 of the aglycone. Cytotoxic activities of two saponins were evaluated against four human tumor cell lines (A549, B16, BEL-7402, and MCF-7) by using the MTT in vitro assay. Compounds 1 and 2 showed moderate cytotoxic activities toward the tested cell lines.

The complete chloroplast genome of Polyspora axillaris (Theaceae).

Polyspora axillaris (Roxb. ex Ker Gawl.) Sweet 1825, is a shrub or tree that is about 9 meters tall in the Theaceae family, mainly distributed in China and Vietnam, and it is widely used as a green tree species in many regions owing to its rapid growth and good adaptability. It is rich in various beneficial extracts for humans, but there are limited studies on it. In this study, we sequenced and annotated the complete plastome of P. axillaris. The chloroplast genome length of P. axillaris is 156,770 bp, with a total of 132 genes, including 37 tRNA genes, 8 rRNA genes and 87 protein-coding genes. The complete chloroplast genome of P. axillaris contains two Inverted Repeats (IRs) of 26,077 bp, a Large Single-Copy (LSC) region of 86,286 bp and a Small Single-Copy (SSC) region of 18,330 bp. The overall G/C content in the chloroplast is 37.3%. Phylogenetic inference shows that P. axillaris formed a sister relationship with P. hainanensis, along with 10 Theaceae species. The research result of P. axillaris will contribute to the genetic preservation of the species and the phylogenetic study of Polyspora.