Comparative genetic diversity and structure of Rhus gall aphid Schlechtendalia chinensis and its host plant Rhus chinensis.

Investigating the population genetic structure of parasites and their host plants can provide valuable insights into their coevolutionary processes. In this study, we assessed and compared the population genetic diversity and structure of 12 Rhus gall aphid (Schlechtendalia chinensis) populations and their respective host plant (Rhus chinensis) using amplified fragment length polymorphism (AFLP) markers. Analysis of molecular variance (AMOVA) revealed that both the aphid and its host plant exhibited higher genetic variance within populations than among them, indicating that their coevolutionary history may have produced analogous patterns of population genetic structure. Additionally, we considered alternative factors that could contribute to this outcome, such as intraspecific gene flow, hybridization, or environmental influences. Our analysis did not reveal a significant correlation between genetic and geographic distances of either the aphid or host plant populations, leading us to reject the isolation-by-distance model as a plausible explanation for the demographic histories of these two species.

Megastigmus seed wasp damage on native Schinus terebinthifolia drupes in ecological restoration area in Brazil.

Megastigmus transvaalensis Hussey (Hymenoptera: Torymidae) parasitizes drupes of Rhus genus plants in Africa and Schinus (Anacardiaceae) in South America. This exotic wasp damages Schinus terebinthifolia Raddi drupes in native forests and ecological restoration areas in Brazil. The objective of the present study was to investigate the precipitation, temperature and relative humidity effects on M. transvaalensis flight activity, and to determine the parasitism rate and sex ratio of this wasp on S. terebinthifolia plants. The study was conducted with yellow sticky traps and S. terebinthifolia drupes collected in an ecological restoration area, from August 2014 to September 2015, in the Sorocaba municipality, São Paulo state, Brazil. Megastigmus transvaalensis populations were negatively correlated with maximum and minimum temperatures and precipitation, with population peaks at the end of May 2015, with 927 insects per evaluation (48.8 adults per trap). The M. transvaalensis sex ratio was higher in the laboratory (0.42) than in the field (0.08). The parasitism rate of S. terebinthifolia drupes by M. transvaalensis ranged from zero to 36.3% under natural environmental conditions. Megastigmus transvaalensis can be monitored with yellow sticky traps. Damage by M. transvaalensis in S. terebinthifolia drupes may decrease the germination of the seeds and the establishment of this plant in native and restoration ecological areas.

Variation and diversification of the microbiome of Schlechtendalia chinensis on two alternate host plants.

Schlechtendalia chinensis, a gall-inducing aphid, has two host plants in its life cycle. Its wintering host is a moss (typically Plagiomnium maximoviczii) and its main host is Rhus chinensis (Sumac), on which it forms galls during the summer. This study investigated bacteria associated with S. chinensis living on the two different host plants by sequencing 16S rRNAs. A total of 183 Operational Taxonomic Units (OTUs) from 50 genera were identified from aphids living on moss, whereas 182 OTUs from 49 genera were found from aphids living in Sumac galls. The most abundant bacterial genus among identified OTUs from aphids feeding on both hosts was Buchnera. Despite similar numbers of OTUs, the composition of bacterial taxa showed significant differences between aphids living on moss and those living on R. chinensis. Specifically, there were 12 OTUs from 5 genera (family) unique to aphids living on moss, and 11 OTUs from 4 genera (family) unique to aphids feeding in galls on R. chinensis. Principal Coordinate Analysis (PCoA) also revealed that bacteria from moss-residing aphids clustered differently from aphids collected from galls. Our results provide a foundation for future analyses on the roles of symbiotic bacteria in plant-aphid interactions in general, and how gall-specific symbionts differ in this respect.

Molecular mechanisms of tannin accumulation in Rhus galls and genes involved in plant-insect interactions.

For galling aphids and their hosts, tannins are crucial for plant-insect interactions and for protecting the host plant from herbivory. Due to their peculiar chemical characteristics, tannins from plant galls have been used for medical and chemical purposes for more than 2000 years. In this study, hydrolyzable tannin concentrations in galls increased from gall initiation (38.34% on June 21) to maturation (74.79% on August 8), then decreased gradually thereafter (58.83% on October 12). We identified a total of 81 genes (named as GTS1-81) with putative roles in gallotannin biosynthesis and 22 genes (TS1-22) in condensed tannin biosynthesis. We determined the expression profiles of these genes by real-time PCR over the course of gall development. Multiple genes encoding 1-beta-D-glucosyl transferases were identified, which may play a vital role in gallotannin accumulation in plant galls. This study is the first attempt to examine the molecular basis for the regulation of tannin accumulation in insect gallnuts. The differentially expressed genes we identified may play important roles in both tannin biosynthesis and plant-insect interactions.

Another look at the phylogenetic relationships and intercontinental biogeography of eastern Asian - North American Rhus gall aphids (Hemiptera: Aphididae: Eriosomatinae): Evidence from mitogenome sequences via genome skimming.

The Rhus gall aphids are sometimes referred to as subtribe Melaphidina (Aphididae: Eriosomatinae: Fordini) and comprise a unique group that forms galls on the primary host plants, Rhus. We examined the evolutionary relationships within the Melaphidina aphids using sequences of the complete mitochondrial genome and with samples of 11 of the 12 recognized species representing all six genera. Bayesian, maximum likelihood and parsimony analyses of the mitochondrial genome data support five well-supported clades within Melaphidina: (1) Nurudea (except N. ibofushi), (2) Schlechtendalia-Nurudea ibofushi, (3) Meitanaphis-Kaburagia, (4) Floraphis, and (5) Melaphis. Nurudea shiraii and N. yanoniella are sister to each other, but N. ibofushi is nested within Schlechtendalia. The Nurudea shiraii-N. yanoniella clade is sister to the large clade of the remaining taxa of Melaphidina aphids. The Bayesian and maximum likelihood analyses support the North American Melaphis rhois as sister to the clade of Floraphis-Kaburagia-Meitanaphis-Schlechtendalia from eastern Asia, whereas the parsimony analysis suggests Melaphis sister to Floraphis with low support (bootstrap support 38%), and the amino acid data weakly place it sister to Schlechtendalia-Nurudea ibofushi. The Melaphis position needs to be further tested with nuclear data. Meitanaphis flavogallis is sister to Kaburagia species instead of grouping with Meitanaphis elongallis. Using the Bayesian method, the North American Melaphis was estimated to have diverged from its closest Asian relatives around 64.6 (95% HPD 59.4-69.8) Ma, which is in the early Paleocene near the Cretaceous and Paleogene boundary (K/Pg boundary). At the K/Pg boundary, mass extinctions caused many types of insect-plant associations to disappear, and these extinctions may explain some of the difficulties in the phylogenetic placement of Melaphis within the analyses.

Protective Effect of Gallotannin-Enriched Extract Isolated from Galla Rhois against CCl4-Induced Hepatotoxicity in ICR Mice.

To investigate the toxicity, protective effects, and action mechanism of gallotannin-enriched extracts isolated from Galla Rhois (GEGR) against carbon tetrachloride (CCl4)-induced hepatotoxicity in Institute for Cancer Research (ICR) mice, alterations in serum biochemical indicators, histopathological structure, antioxidative status, hepatic apoptosis-related proteins, and liver fibrosis regulating factors were measured in mice pretreated with GEGR for five days before CCl4 injection. The GEGR/CCl4 treated group showed decreased levels of three serum marker enzymes (ALP, AST, and ALT) representing liver toxicity, although LDH levels remained constant. Necrotic area indicating hepatic cell death significantly inhibited, while malondialdehyde (MDA) concentration and superoxide dismutase (SOD) expression were dramatically recovered in the GEGR preadministrated group. In mechanism analyses of GEGR, the formation of active caspase-3 and enhancement of Bax/Bcl-2 expression was effectively inhibited in the GEGR/CCl4 treated group. The level of pro-inflammatory cytokines, TNF-α and IL-6, as well as the phosphorylation of p38 and JNK in the TNF-α downstream signaling pathway was rapidly recovered in the GEGR/CCl4 treated group, while anti-inflammatory cytokine (IL-10) increased slightly in the same group. Furthermore, the GEGR/CCl4 treated group showed a significant decrease in collagen accumulation results from alleviation of MMP-2 expression, TGF-ß1 secretion and the phosphorylation of Smad2/3. Taken together, these results suggest that GEGR may induce remarkable protective effects against hepatic injury induced by CCl4 treatment through upregulation of the anti-inflammatory and antioxidant system.

Potential Wound Healing Activities of Galla Rhois in Human Fibroblasts and Keratinocytes.

Wound healing is a complex process orchestrated by the regeneration of the epithelium and the remodeling of the extracellular matrix through processes like collagen deposition. Galla Rhois has been widely used in traditional Korean medicine for its various pharmacological effects, including an anticoccidial effect, however, little is known about its healing activity. The purpose of this study was to determine the effects of Galla Rhois ethanol extract (GRE) on wound healing activities, including H2O2-induced oxidative stress, cell migration, and lactate dehydrogenase (LDH) release assays using human keratinocyte (HaCaT) and dermal fibroblasts (CCD-986SK). In addition, total soluble collagen deposition and collagen gene expression for Type I and III collagen were evaluated in CCD-986SK. Total tannin and flavonoid contents for GRE were measured. GRE induced a significant increase in the number and migration of cells, along with a decrease in cell death and LDH release. In addition, it also induced the over-expression of collagen Type I and III mRNA and caused increased synthesis of total soluble collagen. The contents of total tannin and flavonoid for GRE were 55.7% ([Formula: see text][Formula: see text]mg/g) and 62.9% ([Formula: see text][Formula: see text]mg/g), respectively. The results suggest that GRE can cause accelerated wound healing by increasing cell survival, proliferation, migration, and collagen synthesis along with a potential anti-oxidant property. This evidence provides novel insight into natural therapy for tissue injury.

Melanogenesis inhibition by gallotannins from Chinese galls in B16 mouse melanoma cells.

Nowadays herbal medicines of skin-whitening cosmetics are popular with women. We attempted to find the whitening activity compounds present in many herbal medicines used for this purpose and discuss their mechanisms in melanin biosynthesis. The 70% acetone extracts of 10 kinds of herbs were investigated for their mushroom tyrosinase activity inhibition. Among these 10 extracts, Chinese galls showed inhibitory activity against tyrosinase, with a 50% inhibitory concentration (IC(50)) value of 22 microg/ml. In a B16 mouse melanoma cell culture assay, Chinese galls dose-dependently inhibited melanin biosynthesis. Using ultraviolet A (UVA) or alpha-melanocyte-stimulating hormone (alpha-MSH) to stimulate B16 cells after Chinese gall treatment, the melanin biosynthesis of B16 cells was inhibited in a dose-dependent manner. The active compounds of Chinese galls were isolated by column chromatography, and the melanin biosynthesis inhibition in B16 melanoma cells was measured. Three gallotannins, 2,3,4,6-tetra-O-galloyl-D-glucopyranose, 1,2,3,6-tetra-O-galloyl-beta-D-glucopyranose, and 1,2,3,4,6-penta-O-galloyl-beta-D-glucopyranose, were isolated from Chinese gall extract, and their IC(50) values of tyrosinase inhibition activity were 54, 30, and 15 muM, respectively. By the mushroom tyrosinase inhibition kinetics assay, the three gallotannins were all determined to be non-competitive inhibitors. These results indicated that Chinese galls inhibit melanin biosynthesis, associated with hyperpigmentation and can be used as skin-whitening cosmetics for skin care.

Comparative population structure of Chinese sumac aphid Schlechtendalia chinensis and its primary host-plant Rhus chinensis.

Most of our current understanding of comparative population structure has been come from studies of parasite-host systems, whereas the genetic comparison of gallnut-aphids and their host-plants remain poorly documented. Here, we examined the population genetic structure of the Chinese sumac aphid Schlechtendalia chinensis and its unique primary host-plant Rhus chinensis in a mountainous province in western China using inter-simple sequence repeat (ISSR) markers. Despite being sampled from a mountainous geographic range, analysis of molecular variance (AMOVA) showed that the majority of genetic variation occurred among individuals within populations of both the aphid and its host. The aphid populations were found to be structured similarly to their primary host populations (F (ST) values were 0.239 for the aphid and 0.209 for its host), suggesting that there are similar patterns of gene flow between the populations of the aphid and between populations of its host-plant. The genetic distances (F (ST)/1 - F (ST)) between the aphid populations and between its host-plant populations were uncorrelated, indicating that sites with genetically similar host-plant populations may not always have genetically similar aphid populations. The lack of relationships between genetic and geographical distance matrices suggested that isolation by distance (IBD) played a negligible role at this level. This may be mainly attributed to the founder effect, genetic drift and the relative small spatial scale between populations.

Effect of compounds of Galla chinensis on remineralisation of initial enamel carious lesions in vitro.

OBJECTIVE: To evaluate the effect of compounds of Galla chinensis on the remineralisation of initial enamel carious lesions in vitro. METHODS: Sixty bovine enamel blocks with early lesions were prepared and randomly divided into six treatment groups. The lesions were subjected to a pH-cycling regime for 12 days. Each daily cycle included 4x1min applications with one of six treatments; 1000ppm F aq. (as NaF, positive control); deionized water (negative control); or 4000ppm aqueous solutions of four G. chinensis extracts (GCEs); GCE, GCE-B, GCE-B1, or GCE-B2. Surface enamel microhardness was measured on the enamel blocks before and after demineralisation, and after pH-cycling, and percentage surface microhardness recovery (%SMHR) was calculated. The enamel specimens were then sectioned (thickness ca. 80microm) and examined by polarized light microscopy. RESULTS: All samples rehardened significantly compared to baseline. Fluoride had a significantly greater effect than all other treatments. In the GCEs groups, %SMHR was significantly greater than DDW for the GCE, GCE-B and GCE-B1 groups. There was no significant difference between the GCE-B2 group and DDW. Polarized light microscopy showed that the thickness of the surface layer increased obviously in all specimens including NaF group, GCE group, GCE-B group and GCE-B1 group. Negative birefringent band appeared in the lesions body and the depth of the lesions was obviously reduced. CONCLUSION: The present study has demonstrated the potential of three GCEs (GCE, GCE-B and GCE-B1) to effect net rehardening of artificial carious lesions under dynamic pH-cyclic conditions.

[Study on gallnuts development and its tannin amount].

OBJECTIVE: To find out the suitable time for collecting gallnuts. When Kaburgaia rhusicola parasailed on leaflets of Rhus potainii, the parts of the leaflets developed into gallnuts. METHOD: The regulation of development and tannin amount during whole growing stage were studied. RESULT: K. rhusicola lived in gallnuts for about 80 days. The gallnuts developed quickly during earlier stage and slowly during later stage. Gallnuts maturated from 12th, July to 30th, July, which had no relations with the size and the time for forming, but had relations with development at earlier stage. It developed quickly with earlier maturation and developed slowly with later maturation. Its tannin amount kept stable during the whole growing stage. CONCLUSION: The best harvesting days are those shortly after 7th, July.