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Objects: To evaluate the quality of the root of Ficus hirta Vahl, a pre-column derivation-high performance liquid chromatography (HPLC) method was established for the determination of 16 kinds of amino acids in the root of Ficus hirta from different origins and leaf shapes. Methods: The contents of 16 kinds of amino acids in Ficus hirta were determined by HPLC with phenyl isothiocyanate pre-column derivatization method. The separation was achieved using a ultimate amino acid column (4.6 mm×250 mm, 5 μm) under the gradient elution with the mobile phases composed of 0.1 mol/L sodium acetate buffer and acetonitrile at the flow rate of 1.0 mL/min, column temperature of 40 ℃ and UV detector wavelength at 254 nm. Results: The contents of 16 kinds of amino acids had a good linear relationship, and the correlation coefficients was more than 0.9992, while the average recovery rates (n=6) were 98.1%~105.9% and RSD were 0.67%~2.11%. The average contents of L-asparaginic acid, L-glutamic acid, L-serine, glycine, L-histidine, L-arginine, L-threonine, L-alanine, L-proline, L-tyrosine, L-valine, L-methionine, L-isoleucine, L-leucine, L-phenylalanine, L-lysine were respectively 3.308, 3.424, 2.750, 2.978, 1.302, 4.549, 2.778, 3.989, 3.818, 1.327, 3.732, 0.395, 2.925, 4.549, 2.873, 3.305 mg/g. There were differences in the content of total amino acid in Ficus hirta from different leaf shapes. Compared with leaf shapes, the three-finger shapet had the highest total amino acid, followed by seven-finger shpape, and then five-finger shape, and the last was undivided. The content of total amino acid in Ficus hirta from different producing areas were also different. The highest content was in Lingshan county of Qinzhou city of Guangxi province, followed by Yuannan town of Heyuan city of Guangdong province, and the lowest content was in Chengxi town of Longhai district of Zhangzhou city of Fujian province. Conclusion: 16 kinds of amino acids were separated by this method, which was reliable, accurate and had high recovery rate. It could be used for the determination of amino acids in Ficus hirta, and the content of amino acids could be used to evaluate the quality of Ficus hirta.

Ficus hirta, a well-known medicinal and edible fruit crop widely distributed in the temperate and subtropical areas of China, is rich of flavonoids. However, up to now, the genetic and molecular mechanisms of flavonoid biosynthesis in F. hirta are still unclear. In this study, we assembled the transcriptome of F. hirta using the third-generation sequencing technique single molecule real-time sequencing (SMRT-seq). Then, comparative transcriptome analysis was performed to investigate the fruit gene expression profiling differences among four F. hirta varieties with different lobed leaves (undivided (ud), three-lobed-finger (tf), five-lobed-finger (ff) and seven-lobed-finger (sf)) using RNA-seq. Totally, we obtained 534,375 full-length non-chimeric (FLNC) reads with an average length of 2178 bp by using SMRT-Seq, and 22,782,329, 22,361,639, 21,982,877, and 22,479,690 clean reads for each F. hirta fruit by using RNA-Seq, respectively. Of the assembled 48,399 transcripts, 21,971, 19,183, 21,834, 14,277, 16,255, 16,255 and 21,089 were successfully annotated by NR, SwissProt, KEGG, KOG, Pfam, GO and NT, respectively. And 96 and 21 transcripts were identified to be involved in the phenylpropanoid and flavonoid biosynthesis, respectively. Furthermore, we identified 6, 5, 7, 8, 13 and 14 flavonoid biosynthesis related DEGs in comparison of ud versus tf, ud versus sf, ud versus sf, tf versus ff, tf versus sf and ff versus sf, respectively. And the differential expression of 6 DEGs were confirmed by using quantitative real time PCR. Our study can provide basis for understanding metabolism differences of the bioactive phytochemicals especially flavonoids among different F. hirta varieties. [ABSTRACT FROM AUTHOR]

Abstract Background In brood site pollination mutualisms, pollinators are attracted by odours emitted at anthesis. In Ficus, odours of receptive figs differ among species and the specific pollinators generally only enter figs of their host species ensuring a pre-zygotic barrier to plant interspecific hybridisation. However, field observations recorded that, in Guangdong province in China, Valisia javana hilli, the local pollinator of F. hirta, entered and reproduced successfully in the figs of the closely related F. triloba on a regular basis. We propose that closely related Ficus species produce similar receptive fig odours. Under particular contexts of odours locally present, the receptive fig odours of non-host figs of a Ficus species may become attractive to pollinators of closely related Ficus species. We used the headspace technique to collect in situ receptive fig odours of F. triloba in a series of locations in China. Under controlled conditions, we tested the attraction of fig pollinating wasps from F. hirta and F. triloba to host figs and non-host figs in Y tube experiments. Results Receptive fig odours of F. triloba though different from those of F. hirta, were mainly composed of a same set of volatile organic compounds. When given the choice between receptive fig odours and air, the pollinating wasps were only attracted by their host’s odours. However, when given a choice between host and non-host figs the pollinators of F. hirta were equally attracted by the two odours while the pollinators of F. triloba tended to be more attracted by their host’s fig odours. Conclusions Receptive fig odours vary geographically within species and the differentiation of receptive fig odours between closely related Ficus species is often incomplete. This allows localised or occasional pollinator sharing following different modalities. Cross stimulation when wasps are exposed simultaneously to odours of host and non-host species may be important. While occasional pollinator sharing may play a marginal role when wasp populations are robust, it may ensure the provisioning of new pollinators from the closest relative of a Ficus species if its pollinators go extinct.

Abstract Background Wuzhimaotao (Radix Fici Hirtae) originates from the dry root of Ficus hirta (Moraceae), which is widely known as a medical and edible plant distributed in South China. As the increasing demand for Wuzhimaotao, the wild F. hirta has been extremely reduced during the past years. It is urgent to protect and rationally develop the wild resources of F. hirta for its sustainable utilization. However, a lack of genetic background of F. hirta makes it difficult to plan conservation and breeding strategies for this medical plant. In the present study, a total of 414 accessions of F. hirta from 7 provinces in southern China were evaluated for the population genetics using 9 polymorphic SSR markers. Results A mean of 17.1 alleles per locus was observed. The expected heterozygosity (He) varied from 0.142 to 0.861 (mean = 0.706) in nine SSR loci. High genetic diversity (H e = 0.706, ranged from 0.613 to 0.755) and low genetic differentiation among populations (G’ ST = 0.147) were revealed at population level. In addition, analysis of molecular variance (AMOVA) indicated that the principal molecular variance existed within populations (96.2%) was significantly higher than that among populations (3.8%). Meanwhile, the three kinds of clustering methods analysis (STRUCTURE, PCoA and UPGMA) suggested that the sampled populations were clustered into two main genetic groups (K = 2). Mantel test showed a significant correlation between geographic and genetic distance among populations (R 2 = 0.281, P

One undescribed lignan, one new natural product, along with fourteen known compounds, were isolated from the roots of Ficus hirta. The structures of the isolates were elucidated by comprehensive spectroscopic technologies, including UV, IR, HRESIMS, and NMR. The absolute configuration of 1 was determined by comparison of experimental and calculated ECD data. The cytotoxicity of all the compounds against HeLa and HepG2 cell lines was evaluated and compound 7 showed considerable cytotoxic effect towards HepG2 cells. Also, the apoptotic effect of 7 on HepG2 cells and the effect of 7 on the key proteins (p-JNK and p-p38) in MAPK (Mitogen-activated protein kinases) pathways were studied by flow cytometry and western blotting experiment. As a result, compound 7 induced the apoptosis of HepG2 cells, and dose-dependently increased the phosphorylation of JNK and p38. Thus, 7 might trigger HepG2 cells apoptosis via JNK/p38 MAPK signaling pathway. [ABSTRACT FROM AUTHOR]

Understanding how intraspecies divergence results in speciation has great importance for our knowledge of evolutionary biology. Here we applied population genomics approaches to a fig wasp species (Valisia javana complex sp 1) to reveal its intraspecies differentiation and the underlying evolutionary dynamics. With re-sequencing data, we prove the Hainan Island population (DA) of sp1 genetically differ from the continental ones, then reveal the differed divergence pattern. DA has reduced SNP diversity but a higher proportion of population-specific structural variations (SVs), implying a restricted gene exchange. Based on SNPs, 32 differentiated islands containing 204 genes were detected, along with 1,532 population-specific SVs of DA overlapping 4,141 genes. The gene ontology (GO) enrichment analysis performed on differentiated islands linked to three significant GO terms on a basic metabolism process, with most of the genes failing to enrich. In contrast, population-specific SVs contributed more to the adaptation than the SNPs by linking to 59 terms that are crucial for wasp speciation, such as host reorganization and development regulation. In addition, the generalized dissimilarity modeling confirms the importance of environment difference on the genetic divergence within sp1. Hence, we assume the genetic divergence between DA and the continent due to not only the strait as a geographic barrier, but also adaptation. We reconstruct the demographic history within sp1. DA shares a similar population history with the nearby continental population, suggesting an incomplete divergence. Summarily, our results reveal how geographic barriers and adaptation both influence the genetic divergence at population-level, thereby increasing our knowledge on the potential speciation of non-model organisms.

[Display omitted] • Four undescribed coumarins derivatives, including ficusalt A (1) and ficusalt B (2), and a pair of racemic coumarins, (±) ficudimer A (3a / 3b), were isolated from the roots of Ficus hirta. • Compound 5 had distinct cytotoxic activity against HeLa cells, induced their apoptosis, and significantly inhibited their invasion and migration. • Compound 5 obviously increased the cleavage of PARP, the phosphorylation level of p38 and JNK, and the expression of pro-apoptotic protein of Bax, whereas inhibited the expression of p-ERK and p-AKT. Four undescribed coumarin derivatives, ficusalt A (1) and ficusalt B (2), a pair of racemic coumarins, (±) ficudimer A (3a / 3b), along with ten known amides, were isolated from the roots of Ficus hirta. Their structures were elucidated by several spectroscopic data analyses, including HRESIMS, NMR, and X-ray single-crystal diffraction. The cytotoxic activities of all compounds against HeLa, HepG2, MCF-7, and H460 cell lines were detected using the MTT assay. Among these, 5 showed the highest activity against HeLa cells. Subsequently, the apoptotic, anti-invasive, and anti-migration effects of 5 on HeLa cells were determined by flow cytometer, transwell invasion assay, and wound-healing assay, respectively. The result suggested that 5 distinctly induced the apoptosis in HeLa cells and inhibited their invasion and migration. Further studies on anticancer mechanisms were conducted using Western blotting. As a result, 5 increased the cleavage of PARP and the expression of pro-apoptotic protein Bax. Moreover, 5 notably upregulated the phosphorylation of p38 and JNK, whereas inhibited the expression of p-ERK and p-AKT. Our results demonstrated that 5 could be a potential leading compound for further application in the treatment of cervical cancer. [ABSTRACT FROM AUTHOR]

Abstract Background While the communities constituted by phytophageous insects and their parasites may represent half of all terrestrial animal species, understanding their diversification remains a major challenge. A neglected idea is that geographic phenotypic variation in a host plant may lead to heterogeneous evolutionary responses of the different members of the associated communities. This could result in diversification on a host plant by ecological speciation in some species, leading to geographic variation in community composition. In this study we investigated geographic variation of inflorescence receptacle size in a plant, Ficus hirta, and how the hymenopteran community feeding in the inflorescences has responded. Our predictions were:1)Inflorescence size variation affects wasp species differently depending on how they access oviposition sites.2)In some affected lineages of wasps, we may observe vicariant, parapatric species adapted to different inflorescence sizes. Results We show that fig (the enclosed inflorescence of Ficus) wall thickness varies geographically. The fig-entering pollinating wasp was not affected, while the parasites ovipositing through the fig wall were. Two parapatric species of Philotrypesis, exhibiting strikingly different ovipositor lengths, were recorded. One species of Sycoscapter was also present, and it was restricted, like the shorter-ovipositor Philotrypesis, to the geographic zone where fig walls were thinner. Conclusions Previous work on fig wasps suggested that parapatric geographic ranges among congenerics were due to adaptation to variation in abiotic factors, complemented by interspecific competition. Our results show that parapatric ranges may also result from adaptation to variation in biotic factors. Within an insect community, differences among species in their response to geographic phenotypic variation of their host plant may result in geographically heterogeneous community structure. Such heterogeneity leads to heterogeneous interaction networks among sites. Our results support the hypothesis that plant geographic phenotypic variation can be a driver of diversification in associated insect communities, and can complement other diversification processes.

Endosymbionts living in plants and insects are pervasive. Ficus (Moraceae) has very special inflorescences (which we also call figs) enclosed like an urn, and such inflorescence is usually parasitized by fig wasps. Ficus breeds fig wasp larvae in its figs and adult fig wasps pollinate for Ficus, Ficus and its obligated pollinator formed fig-fig wasp mutualism. Previous studies have found that this confined environment in figs may have provided protection for fig wasps and that this has left some imprints on the genome of fig wasps during the coevolution history of figs and fig wasps. Research on the diversity of both bacteria and fungi in figs are fewer. Our study explored the diversity of endosymbionts in Ficus hirta figs. We utilized high-throughput sequencing and biological database to identify the specific microorganism in figs, then conducted microorganism communities’ diversity analysis and function annotation analysis. As a result, we identified the dominant endosymbionts in figs, mainly some insect internal parasitic bacteria and fungi, plant pathogen, endophytes, and saprotroph. Then we also found bacteria in Ficus hirta figs were more diversified than fungi, and bacteria communities in female figs and functional male figs were different. These findings may give us more insight into the coevolution and interaction among endosymbiont, fig, and fig wasp.

Repeated column chromatography of the n-hexane extract of Ficus hirta leaves (Moraceae) led to isolation of a new oleanane triterpene, 3β-hydroxy-11-oxo-olean-12-enyl-3-stearate (1) in addition to three known compounds, taraxerol (2), 3β-acetoxy-11α-methoxy-12-ursene (3) and 3β-acetoxy-11α-hydroxy-12-ursene (4). Their structures were elucidated by spectroscopic methods and by comparison with data reported in the literatures. [ABSTRACT FROM AUTHOR]

The dry root (Radix Fici Hirtae) of Ficus hirta has been used as a traditional herbal medicine in Ling nan regions of China for a long time. As its large market demand, the wild resources of F. hirta have sharply reduced. It is necessary to conduct the study of conservation genetics. However, there is still lack of complete genome information for the research on evolutionary biology, population genetics and phylogeography of this species. Here, we sequenced the complete chloroplast (CP) genome of F. hirta using Next Generation Sequencing technology (NGS). The CP genome of F. hirta is 160,374 bp in length, which contains a large single-copy (LSC) region of 88,446 bp, a small sing-copy (SSC) region of 18,134 bp, and two inverted repeat (IRa and IRb) regions of 26,897 bp. A total of 130 genes were successfully annotated containing 85 protein-coding genes, 37 tRNA genes and 8 rRNA genes. Phylogenetic analysis support genus Ficus is monophyletic and F. hirta is closely related to F. carica within this genus.

The fruits of Ficus hirta (FH) display strong antifungal activity against Penicillium italicum and Penicillium digitatum. In order to optimize the extraction conditions of antifungal extracts from FH fruit, various extraction parameters, such as ethanol concentration, extraction time, solvent to solid ratio and temperature, were chosen to identify their effects on the diameters of inhibition zones (DIZs) against these two Penicillium molds. Response surface methodology (RSM) was applied to obtain the optimal combination of these parameters. Results showed that the optimal extraction parameters for maximum antifungal activity were: 90% (v/v) ethanol concentration, 65 min extraction time, 31 mL/g solvent to solid ratio and 51 °C temperature. Under the abovementioned extraction conditions, the experimental DIZs values obtained experimentally were 57.17 ± 0.75 and 39.33 ± 0.82 mm, which were very close to the values of 57.26 and 39.29 mm predicted by the model. Further, nine kinds of phytopathogens were tested in vitro to explore the antifungal activity of the FH extracts. It was found for the first time that the FH extracts showed significant inhibition on the growth of P. italicum, A. citri, P. vexans, P. cytosporella and P. digitatum.

The mutualism of figs and their pollinating fig wasps is widely regarded as a model for coevolved mutualism. A high degree of host specificity is ensured by female wasps only being attracted by their specific fig tree species through the volatile organic compounds (VOCs) released by the figs when they are ready to be pollinated. However, very little is known about the molecular mechanisms underlying the production of VOCs and how pollinators respond to these VOCs. Here we present transcriptome sequencing data from VOC-treated fig wasps and control fig wasps. Using Illumina paired-end sequencing, approximately 6.47 Gbp and 6.48 Gbp high quality reads were generated for fig wasps that had been exposed or not to VOCs of their host fig. After read trimming, the de novo assembly of both types of reads produced 58,192 unigenes with an average length of 817 bp. Then functional annotation and GO enrichment analysis was performed by aligning all-unigenes with public protein databases including NR, SwissProt, and KEGG. Differentially expressed genes (DEGs) were investigated using the RPKM method. Overall, 16 up-regulated genes and 13 down-regulated genes were identified. We further performed GO enrichment and metabolic pathway enrichment analyses. One gene involved in the synoptic vesicle cycle and two genes coding for odorant binding proteins (OBP) are likely to have potential impacts on the response of fig wasps to the VOCs emitted by their host figs. This is the first transcriptome sequencing of a fig wasp in the presence of VOCs of its host figs using the next-generation sequencing technology. Our studies suggest that the expression of some genes in the olfactory neural system of the fig wasps is affected by the VOCs released from the figs. This suggests the presence of a dynamic molecular system of detection and hence response to host plant VOCs. As such our findings provide indications for further mechanistic studies on the fig-fig wasp interactions. [ABSTRACT FROM AUTHOR]

Phytochemical investigation of Ficus hirta Vahl. (Moraceae) fruits led to isolate two carboline alkaloids (1 and 2), five sesquiterpenoids/norsesquiterpenoids (3-7), three flavonoids (8-10), and one phenylpropane-1,2-diol (11). Their structures were elucidated by the analysis of their 1D and 2D NMR, and HR-ESI-MS data. All of the isolates were isolated from this species for the first time, while compounds 2, 4-6, and 8-11 were firstly reported from the genus Ficus. Antifungal assay revealed that compound 8 (namely pinocembrin-7-O-β-D-glucoside), a major flavonoid compound present in the ethanol extract of F. hirta fruits, showed good antifungal activity against Penicillium italicum, the phytopathogen of citrus blue mold caused the majority rotten of citrus fruits. [ABSTRACT FROM AUTHOR]

Chemical examination of the liquid culture of Endomelanconiopsis endophytica A326 isolated from the Chinese folk medicine Ficus hirta resulted in the isolation of two new xyloketals named xyloketals K and L (1–2) and three known analogs (3–5) including a new natural product (5). Their structures were determined on the basis of extensive spectroscopic analysis. All compounds were evaluated for their cytotoxic activities against the SF-268, MCF-7, NCI-H460, and HepG-2 tumor cell lines. Nonetheless, no significant activity was observed. [ABSTRACT FROM AUTHOR]

Two new secondary metabolites, endomeketals A–B (1–2), a new natural product (3), and a known compound (4) were isolated from the ethyl acetate extract of the endophytic fungus Endomelanconiopsis endophytica A326 derived from Ficus hirta. Their structures were determined on the basis of extensive spectroscopic analysis. All compounds were evaluated for their cytotoxic activities against SF-268, MCF-7, NCI-H460, and HepG-2 tumor cell lines. However, no compound showed cytotoxic activity against these human tumor cell lines. [ABSTRACT FROM AUTHOR]

Historical abiotic factors such as climatic oscillations and extreme climatic events as well as biotic factors have shaped the structuring of species' genetic diversity. In obligate species-specific mutualisms, the biogeographic histories of the interacting species are tightly linked. This could be particularly true for nuclear genes in the Ficus-pollinating wasp mutualistic association as the insects disperse pollen from their natal tree. In this study, we compare spatial genetic structure of plant and pollinator for the Ficus hirta- Valisia javana association throughout southeast China including Hainan Island, for both nuclear and cytoplasmic markers. We show that dispersal of the insect leads to plant and insect presenting similar signatures of lack of genetic isolation by distance for nuclear genes on the continent over a distance of 1000 km. But we also show that the demographic histories of plant and insect are strikingly different. This is in agreement with extreme climatic events leading to transient regional extinctions of the insects, associated with local survival of the plants. We also observe evidence of genetic differentiation for both wasps and fig-tree between the continent and Hainan Island, although the Qiongzhou Strait is only on average 30 km wide, suggesting that geographic isolation by itself has not been sufficient to generate this differentiation. Hence, our results suggest that in highly dispersive mutualistic systems, isolation-by-dispersal limitation across a geographic barrier could be supplemented by isolation by adaptation, and maybe by coevolution, allowing further genetic divergence. In such systems, species may frequently be composed of a single population. [ABSTRACT FROM AUTHOR]

Background: Root of Ficus hirta (RFH) is widely consumed in China as a plant-derived popular food. However, contents of the active constituents of RFH are unknown, and the chemical as well as bioactive properties of RFH may be affected by growing area. In order to ensure the standard efficacy of health products made with RFH, its active constituents should firstly be determined and, secondly, a means of assessing samples for their contents of these constituents is needed. Results: Four active components, including two coumarins, namely psoralen and bergapten, and two flavonoids, namely luteolin and apigenin, in twenty RFH samples were quantified using a new ultra performance liquid chromatography coupled with photodiode array detector and mass spectrometry (UPLC-PAD-MS) method, and the content level in descending order was psoralen > bergapten > luteolin > apigenin. Chromatographic fingerprint similarity evaluation and cluster analysis were used to assess geographical origin of RFH, and the results revealed a high level of similarity for the tested RFH samples obtained from Hainan, Guangdong, Guangxi provinces and Hong Kong. 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay was conducted to evaluate the antioxidant potencies of the four components, and the results clearly demonstrated that luteolin was most effective; apigenin exhibited a moderate potency, whereas psoralen and bergapten possessed little effect against free radical reactions. Structure-activity relationship of the components was elucidated, and the 3'-hydroxyl group of luteolin was found to be directly responsible for its antioxidant activity. Conclusion: The present UPLC-PAD-MS method and DPPH radical scavenging assay performed well for the purpose of constituent quantification and antioxidant assay. Global profiles were highly similar for RFH samples from different origins. Both the coumarins and flavonoids were involved in the health benefit of RFH. [ABSTRACT FROM AUTHOR]

Background: Wuzhimaotao (Radix Fici Hirtae) originates from the dry root of Ficus hirta (Moraceae), which is widely known as a medical and edible plant distributed in South China. As the increasing demand for Wuzhimaotao, the wild F. hirta has been extremely reduced during the past years. It is urgent to protect and rationally develop the wild resources of F. hirta for its sustainable utilization. However, a lack of genetic background of F. hirta makes it difficult to plan conservation and breeding strategies for this medical plant. In the present study, a total of 414 accessions of F. hirta from 7 provinces in southern China were evaluated for the population genetics using 9 polymorphic SSR markers., Results: A mean of 17.1 alleles per locus was observed. The expected heterozygosity (He) varied from 0.142 to 0.861 (mean = 0.706) in nine SSR loci. High genetic diversity (H e  = 0.706, ranged from 0.613 to 0.755) and low genetic differentiation among populations (G' ST  = 0.147) were revealed at population level. In addition, analysis of molecular variance (AMOVA) indicated that the principal molecular variance existed within populations (96.2%) was significantly higher than that among populations (3.8%). Meanwhile, the three kinds of clustering methods analysis (STRUCTURE, PCoA and UPGMA) suggested that the sampled populations were clustered into two main genetic groups (K = 2). Mantel test showed a significant correlation between geographic and genetic distance among populations (R 2  = 0.281, P < 0.001). Pollen flow, seed flow and/or geographical barriers might be the main factors that formed the current genetic patterns of F. hirta populations., Conclusions: This is a comprehensive study of genetic diversity and population structure of F. hirta in southern China. We revealed the high genetic diversity and low population differentiation in this medicinal plant and clarified the causes of its current genetic patterns. Our study will provide novel insights into the exploitation and conservation strategies for F. hirta., (© 2022. The Author(s).)

A new prenylcoumarin, 5-methoxyl-4,2'-epoxy-3-(4', 5'-dihydroxyphenyl)-linear pyranocoumarin (1), and a new flavonoid, 3-acetyl-3,5,4'-trihydroxy-7-methoxylflavone (2), were isolated from the roots of Ficus hirta. Their structures were elucidated by spectroscopic methods including 1D-, 2D- NMR and HR-ESI-MS. [ABSTRACT FROM AUTHOR]

Endosymbionts living in plants and insects are pervasive. Ficus (Moraceae) has very special inflorescences (which we also call figs) enclosed like an urn, and such inflorescence is usually parasitized by fig wasps. Ficus breeds fig wasp larvae in its figs and adult fig wasps pollinate for Ficus, Ficus and its obligated pollinator formed fig-fig wasp mutualism. Previous studies have found that this confined environment in figs may have provided protection for fig wasps and that this has left some imprints on the genome of fig wasps during the coevolution history of figs and fig wasps. Research on the diversity of both bacteria and fungi in figs are fewer. Our study explored the diversity of endosymbionts in Ficus hirta figs. We utilized high-throughput sequencing and biological database to identify the specific microorganism in figs, then conducted microorganism communities' diversity analysis and function annotation analysis. As a result, we identified the dominant endosymbionts in figs, mainly some insect internal parasitic bacteria and fungi, plant pathogen, endophytes, and saprotroph. Then we also found bacteria in Ficus hirta figs were more diversified than fungi, and bacteria communities in female figs and functional male figs were different. These findings may give us more insight into the coevolution and interaction among endosymbiont, fig, and fig wasp. [ABSTRACT FROM AUTHOR]

The dry root (Radix Fici Hirtae) of Ficus hirta has been used as a traditional herbal medicine in Ling nan regions of China for a long time. As its large market demand, the wild resources of F. hirta have sharply reduced. It is necessary to conduct the study of conservation genetics. However, there is still lack of complete genome information for the research on evolutionary biology, population genetics and phylogeography of this species. Here, we sequenced the complete chloroplast (CP) genome of F. hirta using Next Generation Sequencing technology (NGS). The CP genome of F. hirta is 160,374 bp in length, which contains a large single-copy (LSC) region of 88,446 bp, a small sing-copy (SSC) region of 18,134 bp, and two inverted repeat (IRa and IRb) regions of 26,897 bp. A total of 130 genes were successfully annotated containing 85 protein-coding genes, 37 tRNA genes and 8 rRNA genes. Phylogenetic analysis support genus Ficus is monophyletic and F. hirta is closely related to F. carica within this genus. [ABSTRACT FROM AUTHOR]

The active ingredients of Ficus hirta and Hypericum perforatum were collected from Traditional Chinese Medicine Database and Analysis Platform(TCMSP) and related papers. The potential targets of these two medicinal herbs were searched from HERB database, and those associated with microvascular angina were screened out from GeneCards, Online Mendelian Inheritance in Man(OMIM), Therapeutic Target Database(TTD), and HERB. Cytoscape was used to construct a protein-protein interaction(PPI) network of the common targets shared by the two herbs and microvascular angina based on the data of String platform. Metascape was employed to identify the involved biological processes and pathways enriched with the common targets. Cytoscape was used to draw the &quot;active ingredient-target-pathway&quot; network. AutoDock Vina was used to dock the core ingredients with the key targets. A total of 19 potential active ingredients and 71 potential targets were identified to be associated with microvascular angina. Bioinformatics analysis showed that phosphatidylinositol-3-kinase/protein kinase B(PI3 K-AKT), interleukin-17(IL17), hypoxia-inducible factor 1(HIF-1) and other signaling pathways were related to the treatment of microvascular angina by F. hirta and H. perforatum. Molecular docking results showed that β-sitosterol, luteolin and other ingredients had strong affinity with multiple targets including mitogen-associated protein kinase 1(MAPK1), epidermal growth factor receptor(EGFR) and so on. These findings indicated that F. hirta and H. perforatum may regulate PI3 K-AKT, IL17, HIF-1 and other signaling pathways by acting on multiple targets to alleviate oxidative stress, inhibit inflammatory response, regulate angiogenesis, and improve vascular endothelium and other functions. This study provides reference for in vitro and in vivo studies of the treatment of microvascular angina.

[Display omitted] • A pair of undescribed enantiomers (1a / 1b) was isolated from the roots of Ficus hirta. • Compound 4 showed enhanced cytotoxicity on HeLa, MCF-7 and H460 cells. • Compound 4 induced HeLa cells apoptosis and increased the PARP cleavage. • Compound 4 increased the expression of p-JNK and p-p38. • Compound 4 reduced the protein levels of p-AKT and p-ERK. A pair of undescribed enantiomers, (±) ficusflavonid A (1a / 1b), along with five known analogues, were isolated from the roots of Ficus hirta. Their structures were determined by the analysis of extensive spectroscopic data (including UV, IR, HRESIMS and NMR). Two enantiomers (1a and 1b) were successfully separated by chiral chromatographic column and their absolute configurations were assigned by the comparison of experimental and calculated ECD data. The cytotoxicity of all the isolates against HeLa, MCF-7, HepG2 and H460 cell lines were evaluated by MTT assay. Among them, 4 suppressed the proliferation of HeLa cells with the IC 50 value of 28.88 μM. Furthermore, the apoptotic effect of 4 on HeLa cells and the level of several crucial proteins in AKT/MAPKs signaling pathways were analyzed by flow cytometer and western blot assay. As a result, 4 induced HeLa cell apoptosis in a dose dependent manner and significantly increased the protein levels of p-JNK and p-p38, whereas distinctly reduced the expression of p-AKT, and p-ERK. Thus, compound 4 might induce HeLa cells apoptosis via MAPK and AKT signaling pathways, which could be considered as a potential leading compound for the development of anticancer drugs. [ABSTRACT FROM AUTHOR]