Northwestern Amazonia is home to a great degree of linguistic diversity, and the human societies in that region are part of complex networks of interaction that predate the arrival of Europeans. This study investigates the population and language contact dynamics between two languages found within this region, Yukuna and Tanimuka, which belong to the Arawakan and Tukanoan language families, respectively. We use evidence from linguistics, ethnohistory, ethnography and population genetics to provide new insights into the contact dynamics between these and other human groups in NWA. Our results show that the interaction between these groups intensified in the last 500 years, to the point that it is difficult to differentiate between them genetically. However, this close interaction has led to more substantial contact-induced language changes in Tanimuka than in Yukuna, consistent with a scenario of language shift and asymmetrical power relations. This At the centre of this large area, social exogamy and multilingualism create a marriage practice known as linguistic exogamy, in which marriages are forged between members of different patrilineal descent groups which are associated with different languages. These marriages bring together speakers of several Eastern Tukanoan (ET) and Arawakan languages, as well as the Carijona language of the Cariban family  –7. LinguThis paper examines the population and language contact dynamics between two languages found within this region, Yukuna and Tanimuka, which belong to the Arawakan and Tukanoan language families, respectively. Yukuna is the identity language of the Yukuna and Matapi groups, who number approximately 700–1000 people ; TanimukGiven that contact and admixture among human populations has been a constant throughout history ,16, globExtensive contact in the context of intermarriage can be expected to lead to different genetic patterns depending on sociocultural factors such as marriage practices and postmarital residence rules. For instance, in the absence of asymmetrical power relations or strong rules for endogamous versus exogamous marriage among the groups in contact, one would expect to find relatively few genetic differences. However, it has been shown that among small-scale human societies, intermarriage is often sex-biased, with a preference toward one of the sexes marrying into the other group. Since patrilocality is a frequent postmarital residence pattern among human societies, it is common to find women moving to the husband's ancestral territory –32. FurtIn this study, we offer novel insights into the population history and the dynamics of language change in NWA by bringing together evidence from linguistics, ethnohistory, ethnography and population genetics regarding Yukuna and Tanimuka, as well as their place within the broader NWA social panorama. Of course, there are important differences in the way culture, language and genes are transmitted and changed, and differences in the time scales on which these processes operate (for discussion see –40). The. 2To learn more about the contact situation between these groups, we adopt a multidisciplinary approach involving linguistic, sociocultural and genetic databases. Each of these types of data informs us about different sociocultural, demographic and historical aspects of the human societies of Northwestern Amazonia. Here we describe the kinds of data and the analytical approaches used for each type of evidence.. 2.1We collected previously reported genetic data from several indigenous groups from NWA , includia priori-defined number of ‘K’ ancestral source populations [K = 2 to K = 14 with 20 replicates for each K and used the cross-validation error procedure to find the ‘K’ that best describes the genotype data. We visualized ADMIXTURE results with the software pong [We used two analytical approaches with the genome-wide data. The first one uses allele-frequency-based methods and includes PCA (as implemented in the Eigensoft package ), model-ulations . We ran are pong .2-statistics, outgroup-f3-statistics, f4-statistics and f4-ratios . We ran SHAPEIT with options –burn 10, –prune 10 and –main 30 for iteration number with 500 conditioning states, leaving other parameters as default as previously reported [https://github.com/dangliu/Massim_project.The second approach uses haplotype-based methods, that is, long and uninterrupted blocks of DNA inherited from a set of common ancestors . We perfreported , which rreported . In addireported ,20. The With the uniparental data we focused on reconstructing haplotype networks, implemented in the R-package pegas v. 1.1 , using t. 2.2We started by compiling a large set of sociocultural data from ethnolinguistic groups in Western South America , designed to address a broad range of questions regarding the population history of this part of the continent. We subset this database to match the ethnolinguistic groups described in For this paper, given the importance of gender-specific sociocultural practices in NWA, we chose to separately analyse practices generally transmitted from fathers to sons, and those generally transmitted from mothers to daughters .  meaning, and grammatical contexts of noun classification marking strategies. 4.Case marking: the form and meaning of case markers. 5.Tense, aspect, modality, evidentiality marking: the form and meaning of markers that code temporal, aspectual, modal or evidential information. 6.NP syntax: the relative order of elements and morphosyntactic marking patterns of the noun phrase. 7.Clausal syntax: the relative order of elements and morphosyntactic marking patterns of the clause.We consulted written sources to collect data on the grammars of 18 NWA languages belonging to the Tukanoan and Arawakan language families . We collconstructions, using the term broadly to mean a recurring syntagmatic pattern of language that pairs a formal realization to an interpretation (form-meaning pair). It is a broad use of the term in the sense that it can be interpreted abstractly, as in the case of phonemes and allophones, for which it can be said that allophones are the formal realization (form) of a phoneme .These data have all been conceived as inventories that languages may have . We call the observational units for each of these inventories . 2.4After we standardized the databases described in §§ 2.2 and 2.3 using the glottospace R package , we calc. 3. 3.1a). Variation in PC1 separates speakers of Guahiban languages on the right from Yukuna and Nukak individuals on the left; Piapoco individuals form a cline towards the Guahiban-speaking groups, reflecting some genetic similarities with them. PC2 separates Nukak individuals at the bottom-left and a cluster of Yukuna, Matapi and Tanimuka individuals on the top-left of the plot. We observe that five out of six Tanimuka individuals cluster together with Yukuna and only one Tanimuka individual appears closer to a cluster formed by the rest of Eastern-Tukanoan (ET) speaking groups, Curripaco, four Piapoco individuals from the HGDP panel [Figure 1b shows the results for the K ancestral populations with the lowest cross-validation error . We provide the full result (i.e. K = 2–14) in the supplementary materials . In figure 1b we observe that Yukuna, Matapi and Tanimuka are assigned a similar ancestry profile. By contrast, other ET-speaking groups, the language family to which Tanimuka's language belongs, are assigned a mixture of ancestries. However, the ancestry component that is maximized in Yukuna, Matapi, and Tanimuka (blue ancestry) represents on average 46% of the ancestral components assigned to ETs (figure 1b).We used principal component analysis (PCA) to visualize the broad patterns of genetic variation among individuals and groups included in this study a. VariatDP panel  and Puinb). Pairwise f2-distances show the existence of several clusters showing high genetic similarities . This cFurthermore, we used a f4-statistic of the form f4 , where NAmong those groups, Carijona shows the highest positive f4-value. This result is consistent with oral histories of groups from the Apaporis and Mirití-Parana Rivers, who attribute a common origin to the Tanimuka and the Carijona ,88.We estimated admixture proportions in Tanimuka using an f4-ratio ; based ox-axis and recipient populations on the y-axis. Although Yukuna and Matapi appear as donors of ancestry to Tanimuka, it is the Curripaco who contribute more ancestry to Tanimuka, as well as to Matapi and Yukuna. By contrast, ET groups contribute less ancestry to Tanimuka than they do to Yukuna or Matapi. Also, Matapi individuals are recipients of diverse ancestries, since we observe consistent high mean values with multiple groups in NWA, the top donors being Murui, Puinave and Curripaco. However, as has been shown by Hellenthal et al. [We then investigated whether the observed genetic affinity patterns based on the correlations of allele frequencies were attributable to recent demographic events. For this, we used haplotype-based methods that inform us about events which occurred on the order of tens of generations to a couple of hundred generations up to the present ,73. Partl et al. , if a sol et al. .Figure Moreover, patterns of IBD sharing  show that Tanimuka, Yukuna and Matapi share a large number of very long IBD blocks both within and between them, indicating that these three groups have a large number of common ancestors during the last 500 years, a likely consequence of their extensive and continuous genetic admixture through this time period.. 3.1.1a,b) shows differences in how Tanimuka and Yukuna relate to each other and to other groups in NWA. For the mtDNA Yukuna seem to have wider connections to other groups than Tanimuka, as Yukuna show more shared and related haplotypes with different ethnolinguistic groups. Tanimuka mtDNA and Y-chromosome haplotypes are mainly shared with or closely related to haplotypes in the Yukuna, with just a few exceptions that show links with haplotypes in the Barasana, Curripaco and Desano and Tukano.Uniparental data , ‘a 1.Are Arawakan constructional profiles significantly different from Tukanoan ones? 2.Are Tanimuka and Yukuna significantly different from the respective family profiles? 3.Are Tanimuka and Yukuna significantly different from each other?Our approach to the matter of language shift in linguistics is as follows: on the basis of linguistic distances between constructions (see §2.4), we can assess to what extent Tanimuka and Yukuna are more similar than expected (i.e. not significantly different), and to what extent Tanimuka and Yukuna differ from the corresponding trends in their respective language families. We, therefore, focus on the following three questions:1From these three questions, we can schematically represent six possible patterns, shown in 2 and Tanimuka and Yukuna themselves are not significantly different from each other (Q3 = N). An asymmetrical diffusion pattern holds if Arawakan and Tukanoan are significantly different (Q1 = Y), one of the languages is different (Q2 = Y) while the other is not (Q2 = N), and Tanimuka and Yukuna are not significantly different (Q3 = N).The historical scenarios that we focus on in this paper are convergence and language shift. These two scenarios would be compatible with patterns d and f, respectively. Scenario d (symmetric diffusion) can be identified if the Arawakan and Tukanoan profiles are significantly different from each other (Q1 = Y), Tanimuka and Yukuna are different from their respective family profiles (Q2 = Y for both languages),The typical outcomes and characteristics of convergence and shift are summarized in f) is not straightforward. The presence of pattern f can be a false positive for a historical scenario of shift, and the absence of pattern f can be a false negative. The reason for the former (pattern f is present but not indicative of shift) is because scenario f is also compatible with a situation of language maintenance, in which there is strong inequality in power relations and/or unidirectional bilingualism. Nor does the absence of pattern f exclude a shift scenario.Note, however, that the connection of (in particular) language shift  to pattefigure 5. 3.2.1Of the seven grammatical domains we investigated, three are consistent with a shift scenario. These are classifiers, person markers, and tense-aspect-modality-evidentiality (TAME). The latter shows this pattern most clearly, so we discuss it here. We tested this visual pattern by applying a permanova test, where we contrasted Arawakan (without Yukuna), Tukanoan (without Tanimuka), Yukuna and Tanimuka. The results are given in As can be seen in p = 0.012), but where Tanimuka is similar to Arawakan (p = 0.45), Yukuna is still significantly different from Tukanoan (p = 0.006), and Tanimuka and Yukuna show non-significant differences (p = 0.126).The other two areas of grammar where we find asymmetrical patterns in which Tanimuka has become Arawakanized, but Yukuna has not become Tukanoanized (or at least much less so) are classifiers and person markers. Nevertheless, the patterns are slightly different for these two datasets . The results for person markers are similar to those of TAME, except that the difference between Tanimuka and Arawakan is also non-significant. The classifiers show a slightly weaker asymmetrical pattern in that Yukuna is also significantly different from its Arawakan relatives . The phoneme-allophone database shows a pattern in which Tanimuka is both similar to Tukano (p = 1) and Arawak (p = 0.084), while Yukuna is different from both (p = 0.006 for both), suggesting a possible change within Tanimuka toward the Arawakan profile. The last two datasets show different patterns. Case shows parallel structures throughout the two families, including Tanimuka and Yukuna, as none of the groups are significantly different from each other. The only dataset that shows signs of convergence is the dataset on clausal structure, where Yukuna and Tanimuka are not significantly different from Tukanoan and Arawakan, respectively, and from each other .Of the remaining four datasets, two show weak asymmetric patterns. Noun phrase structure is significantly different for all pairs, except for Yukuna and its Arawakan sister languages, but Tanimuka does seem to have become more similar to (though still significantly different from) Yukuna . This is the pattern that we might expect if, as suggested by , the AraAs discussed in §2.2 and The NMDS plot in et al. [Unfortunately, subsetting the dataset to reflect only cultural practices transmitted from mothers to daughters presented some problems in the analysis. In particular, the high degree of homogeneity in women's cultural practices in the NWA  with our focal groups. Ethnohistorical accounts as well as the oral traditions of groups from the Mirití-Parana/Apaporis Rivers are consistent with this picture, suggesting that the Tanimuka arrived relatively recently to this area , perhapsJerí. Similar accounts are found among the Makuna, an ET-speaking group from the Apaporis River, who call the shared jaguar ancestor Yainakahí. However, the Tanimuka do not agree, and say instead that Yainakahí is only the ancestor of the Carijona and the non-indigenous Colombians [Furthermore, genetic data are also consistent with oral histories and mythology. For instance, according to Yukuna oral histories, the Tanimuka, the Carijona, and white people have the same mythical origin, the jaguar lombians ,88,96. Alombians  and the lombians , relativprototypical shift and prototypical convergence scenarios. There are many ways in which actual social scenarios can differ from the ones shown in this figure, and the theoretical opposition between convergence and shift is best regarded as a continuum. In particular, the historical effects of language shift can be all but absent under certain circumstances. Notably, if there is child bilingualism, with full access to L2 (the target language), shift may leave no trace of the original language. In addition, effects may be differential across different parts of the linguistic system. These issues make it difficult to predict outcomes of contact scenarios, and thus to reconstruct social scenarios from contemporary data. In most cases, a reconstruction can suggest a most likely scenario, and even for that, linguistic data need to be combined with data from other disciplines to complete the reconstruction of the social context.Linguistic data provide evidence of the pervasive contact influences on the Yukuna and Tanimuka languages. We observe signals of both convergence and asymmetrical impact on their languages, in which Tanimuka has been influenced by Yukuna much more than vice versa. This can be seen in Although there is no way of reconstructing a historical language shift that has left no trace in the linguistic data, we can  distinguish between maintenance and shift scenarios and thus reduce the risk of a false positive. In their aforementioned influential model of language contact and its outcomes, Thomason and Kaufman  make a bWanderwörter from Tupian, Cariban and unidentified sources. The cognates that are exclusively shared between Yukuna and Tanimuka-Letuama are suggestive of bilateral and historically recent contacts, not of long-term, intensive contact resulting in large amounts of borrowed lexicon . This reduces the likelihood of a maintenance scenario and thus a false positive in cases of asymmetrical diffusion  share 32 each with a common etymology. This corresponds to 70% of all 46 loans found in the Tanimuka-Letuama lexicon. From this, 9 are exclusively shared between Yukuna and Tanimuka. Among these, 6 are loans from Yukuna into Tanimuka-Letuama, while 3 have an unclear directionality. For the remaining 37 etyma in Tanimuka, 24 come from an Arawakan language other than Yukuna, and 13 are Similar signals are observed in the genetic data. f4-statistics show that, although Tanimuka is closely related to Yukuna, Tanimuka has additional signals of admixture with Carijona, ET groups and Nukak. However, our genetic analyses suggest that the genetic history of human groups in NWA is very complex, with individuals and groups showing genetic similarities to several other groups, irrespective of linguistic and cultural differences (e.g. figures Furthermore, uniparental genetic data suggest that these interactions were different for men and women. We observed that on the male-specific side, Yukuna and Tanimuka's Y-chromosome haplotypes are closely related and different from other Arawakan and ET haplotypes. By contrast, on the maternal side we observed that the Yukuna exhibit more haplotypes and these are related to haplotypes from different ethnolinguistic groups, while Tanimuka's mtDNA haplotypes are mainly shared with Yukuna or closely related to Yukuna haplotypes. This might suggest that Yukuna has been involved in a larger network of interactions with other ethnolinguistic groups in NWA, while the Tanimuka have primarily interacted with the Yukuna. However, this could be due to the large differences in sample size for the uniparental data . The Y-cIn conclusion, we cannot clearly reject either of the hypotheses proposed in this study, since the evidence available to us shows signals that are consistent both with language shift and convergence. Therefore, we argue that it is likely that both processes have operated through the long history of contact between Yukuna and Tanimuka and more broadly between Arawakan and Tukanoan-speaking groups in NWA. However, our data clearly show that Tanimuka and Yukuna look genetically more similar to each other than to their respective language family members and that the Tanimuka language has been influenced by Yukuna much more than vice versa, and although both signals are consistent with language shift, it does not rule out convergence with asymmetrical power relations. Genetically, Arawakan-speaking groups do not show strong genetic similarities to each other, with the exception of our sample of Piapoco and Achagua who live side-by-side. Instead, Arawakan groups tend to show genetic affinities with their non-Arawakan geographical neighbours, which is consistent with the idea that Arawakan groups were pivotal in maintaining regional systems of exchange with diverse ethnolinguistic groups, in which multilingualism and intermarriage were common features ,99,100.


Solanum lycopersicum L.) genetic basis of traits conferring HMs tolerance and yield are assessed when irrigated with waste or canal water. The North Carolina Mating II analysis illustrate the amount of gene action, nature, and inheritance pattern. Genetic components depict the involvement of non-additive, additive, and maternal genetic effects in HMs tolerance inheritance and yield. A noticeable increase in cumulative additive variance for the number of flowers  and the number of fruits  is recorded for tomato plants irrigated with wastewater, illustrating additive gene action. However, female and male (MSf/MSm) square ratios also show an association with cytoplasmic inheritance. For HMs tolerance, both additive and dominant variances appeared to be significant; cumulative dominance variance  is higher compared to additive variance  for nickel (Ni), chromium (Cr), lead (Pb), manganese (Mn), and zinc (Zn), respectively, indicating dominance gene action. The genotype RIOGRANDI accumulated and translocated fewer HMs to the aerial part of the plant compared to CLN-2418A and PB-017906, thus presenting a tolerant tomato genotype according to the hydroponic experiment. This also exhibited a differential pattern of gene action for HMs tolerance, suggesting that genotypes possess significant differences for HMs tolerance.Wastewater irrigation is a substitute for surface water scarcity, but traces of heavy metals (HMs) result in deleterious implications for soil, crop productivity, and in humans. Crops presenting HMs tolerance in genetic behavior are important for producing tolerant genotypes cultivated under wastewater irrigation. In the first part of this experiment, the results obtained previously are re-assessed in a hydroponic system and similar patterns and concentrations of HMs are found in different tomato organs. Following this trial, the tomato’s ( Along with additive and non-additive gene effects, maternal effects were also involved in controlling the number of flowers, the number of fruits, and HMs tolerance, indicating the interference of the cytoplasm. HM Involvement of maternal effects for multiple HM tolerance through the MTP gene family has also previously been reported by El-Sappah et al. 1/2 showed over-dominance for the control of HMs tolerance, indicating that both additive and non-additive genetic effects control HMs tolerance.Heritability is considered an excellent index for transmitting characteristics to subsequent generations. To realize improvement in yield-contributing traits, a plant breeder must assess the percentage of inheritance in a population for any particular trait. It also enlightens the efficiency of subsequent generations regarding the selected traits in any breeding experiment. Genotypic, as well as environmental variance, can be assumed to approximate the chances of heritability and genetic advance, which depicts the probability of genetypic differences concerning environmental variables. Heritability and genetic advance were high with high additive gene effects for the number of flowers and fruits; therefore, direct selection without hybridization may help improve these characteristics . These rHeritability and genetic advance estimates also provide information about yield and HM tolerance inheritance patterns. Traits possessing higher genetic advances with high or low heritability estimates are desirable because of their higher additive genetic effects; therefore, selection may be effective based on genetic advances. Higher heritability estimates with lower genetic advances are undesirable because of non-additive genetic effects, which show that environmental effects regulate trait expression. Heritability and genetic advance were high to moderate for Pb, Cr, Mn, and Ni tolerance but not Zn. Therefore, direct selection without hybridization may be helpful for these characteristics . BecauseEstimates for different genetic components revealed that an additive genetic effect was responsible for the inheritance of yield-related traits such as the number of flowers and the number of fruits per plant. In contrast, a dominant genetic effect was responsible for the inheritance of different HMs tolerance. From the above results, it was observed that along with additive and non-additive genetic effects, maternal effects were involved in controlling different traits such as yield and HMs tolerance in tomatoes, showing the involvement of cytoplasm. An evaluation of the many hybrids/accessions is recommended to develop low metal-accumulator tomato genotypes with a higher yield. Because the above results are from a limited set of tomato germplasm, they cannot be generalized to all tomato genotypes . Multiple genes influenced genetic variation for metal tolerance with both additive and non-additive effects, but for HM tolerance, dominant genetic effects were observed most commonly , which a3 for 3 h under lab conditions. The samples were heated at 250 °C on a heating block until the appearance of tinted yellow color of the solution in digestion flask. The solution was diluted with distilled water up to 50 mL and filtered for the analysis of HMs with atomic absorption spectrophotometer. Two HM tolerant/low metal-accumulator genotypes (PB-017906 and CLN-2418A) and one susceptible/high metal accumulator genotype (RIOGRANDI) were selected out of 44 genotypes in a screening trial based on HM accumulation in a field experiment  and then results were re-assessed in the hydroponic experiment. A similar pattern of HM concentrations “as in sewage water”  was mainSeeds of the selected tomato genotypes were sown in sand-filled seedling trays, and moisture content was maintained. After 25 days of seedling establishment, in November 2018, seedlings were transplanted to the field soil in a replicated trial under two treatments, i.e., wastewater and canal water. At maturity, the crossing was performed according to North Carolina Design-II matting to develop genetic material for assessing genetic components of variability. Analysis of water and soil used in current experiment given in Floral buds of all genotypes in both water treatments were covered with a butter paper bag and emasculated with forceps two days before blooming. Any opened flowers present on the same inflorescence were removed to avoid mating. Desired male flowers of all the genotypes in both water treatments were stored in clean plastic bottles, and anthers were collected and sun-dried, so they opened to release pollens for crossing. Pollens were applied to exposed stigma with a dissecting needle; to ensure a good seed set, the style was repeatedly dipped in a pollen-containing petri dish.At maturity, fruits of F1 and selected parents were collected in bags, crushed, and fermented. Seeds were washed with fresh water to remove flesh and skin. Floating pieces were removed gently, and washing was repeated many times by adding fresh water to remove flesh and gel. Clean seeds were collected, dried, and stored. 1 hybrids seeds were sown in seedling trays and transplanted in field after 25 days of the seedling establishment during November 2019 in a triplicated trial in two sets, i.e., canal water and wastewater. Data regarding HM and yield-related traits were recorded at maturity by harvesting and analyzing ripened fruit. Accumulation of nickel (Ni), zinc (Zn), manganese (Mn), chromium (Cr), and lead (Pb) in tomato fruit were determined by performing acid digestion as described by [To assess inheritance patterns and genetic studies for HMs tolerance and yield-related traits, HMs tolerant (PB-017906 and CLN-2418A), susceptible parental genotypes (RIOGRANDI), and Fribed by . For yieBS) and genetic advance (GA) were calculated using formulas [Data were analyzed statistically using the analysis of variance technique  to find formulas ,59. Valuformulas . Data weformulas .Under such circumstances of surface water scarcity, wastewater is used for irrigation purposes, but one of its drawbacks is contamination with HMs. Due to the high cost, treatment is not possible in underdeveloped countries such as Pakistan. One of the solutions to this problem is to develop genetically tolerant genotypes that hold genetic capability physiologically and possess molecular mechanisms to block HMs from entering the food chain. Genotypic variation exhibited a differential pattern of gene action for HMs tolerance, suggesting that genotypes possess significant differences for HMs tolerance. Since roots are the first organ for the hyperaccumulation of different HMs ions as shown in different tomato genotypes, particularly RIOGRANDI, therefore, the pattern of enzyme activities and other metal accumulation in the root system needs to be determined, and intermittent use of clean water between wastewater irrigation will not only reduce the heavy metal load but could also enhance the soil fertility.


Background and Objectives: Non-cancer chronic pain (CP) results from the interaction between genetic and environmental factors. Twin studies help to estimate genetic and environmental contributions to complex traits such as CP. To date, twin studies on the heritability of pain phenotypes have relied almost exclusively on specific diagnoses, neglecting pain intensity. This study aims to estimate the genetic and environmental contributions to CP occurrence as a wide phenotype and its intensity among a non-clinical population. Materials and Methods: A nationwide online survey was conducted in February 2020 on 6000 adult twins enrolled in the Italian Twin Registry. A five-item questionnaire, designed and validated by our study group, was administered to detect the CP condition along with its intensity, underlying causes or triggers, treatments, and self-perceived efficacy. The twin study design was used to infer the relative weight of genes and environment on CP occurrence and intensity, and biometrical modelling was applied to these phenotypes. Results: A total of 3258 twins, aged ≥18, replied to the online survey (response rate 54%). These included 762 intact pairs , of whom 750 pairs were subjected to biometrical modelling after the exclusion of pairs with either unknown zygosity or cancer-associated CP. Broad-sense heritability estimates were driven by non-additive genetic effects and were 0.36 (0.19–0.51) for CP occurrence and 0.31 (0.16–0.44) for CP intensity. No evidence emerged for either sex differences in genetic and environmental variance components or interactions of these components with age. Conclusions: Moderate non-additive genetic components were suggested for non-cancer CP occurrence and its intensity. These results encourage further research on the gene–gene interactions underlying CP liability and associated phenotypes, and also strengthen the need for prevention strategies to avoid CP occurrence or to decrease pain intensity. Non-cancer CP (referred to as CP in this work), i.e., pain persisting or recurring for more than three months , affectsDespite the fact that CP may actually refer to different phenotypes and diagnostic entities recently categorized within the International Classification of Diseases-11 (ICD-11) , there aIn this framework, the main aim of the present study is to estimate the genetic and environmental components of CP occurrence as a wide phenotype (irrespective of underlying causes/triggers) and its intensity, as well as to explore the possible sex- and age-moderation of these components in a sample of adult twins enrolled in the Italian Twin Registry (ITR) of the Istituto Superiore di Sanità.The present study was approved by the Ethics Board of the Istituto Superiore di Sanità on 14 April 2020. Participants underwent an informed consent procedure.A nationwide online survey, using the LimeSurvey Professional Platform , was conducted in February 2020 on 6000 MZ and DZ adult twins of Caucasian origin previously enrolled in the population-based ITR . CurrentTwin pairs were classified as MZ or DZ based on standard questions on physical resemblance and frequency of confusion of the twins by family members and strangers . This isThe Brief Five-Item Chronic Pain Questionnaire was used for CP assessment. It is a validated brief self-administered measure, particularly suitable to detect persistent states of pain and related characteristics in general population surveys . It is cThe socio-demographic and CP-related characteristics of the study sample were summarized using means (with standard deviations and ranges) for continuous variables and percentages for categorical variables. Descriptive analyses were performed by the Stata software version 16 .Biometrical structural equation model-fitting analyses were conducted, in accordance with the assumptions of the twin design, to estimate the heritability and the environmental component of CP occurrence and intensity.For the analysis of CP intensity, the following scale was considered: 0 = no pain, 1 = very mild/mild, 2 = moderate, 3 = severe/very severe, with the “very mild” and “very severe” responses of the original scale combined, respectively, with the “mild” and “severe” ones due to low numbers. As in other commonly used scales of pain intensity , the numerical rating scale (NRS)), two endpoints from “no pain” to “very severe pain” were considered.Given the dichotomous and ordinal nature of occurrence and intensity respectively, liability-threshold models were applied . Using tAmong the 6000 invited twins, 3258 replied to the online survey (response rate 54%). This response rate was substantially higher than that generally observed in previous ITR surveys. No significant differences emerged between the respondents and non-respondents with respect to the main socio-demographic characteristics . Among the 3258 twins, there were 1524 twins from 762 intact pairs, plus 1734 unmatched twins. Given that correlation and model-fitting analyses of twin data mainly exploit within-pair information, only twins from the 762 intact pairs were considered. No major differences were detected between twins from intact pairs and unmatched twins with respect to zygosity or to socio-demographic and CP-related profiles. All 762 pairs were included in the descriptive analysis, while 750 pairs included in the correlation analysis and structural equation modelling, after the exclusion of 7 pairs with unknown zygosity and 5 pairs for which CP occurrence was declared to be associated with a tumor diagnosis.The main socio-demographic and CP-related characteristics are summarized in The mean age was 39 years (range 18–82 years), 34% of the twins were male, and 57% were MZ. Their education level was medium–high, with more than 50% of subjects having at least a three-year university degree. CP prevalence was about 24%, with 60% of CP-affected subjects reporting a moderate pain intensity and 18% declaring a severe or very severe pain. As regards the causes of pain, trauma without surgery was the most reported origin (40%), followed by “another illness diagnosed by a doctor” (35%) and a “poorly defined disease” (21%). Among twins suffering from CP, 74% reported being under  treatment, and of these, 86% declared that they had received beneficial medication effects.The results of the correlation and model-fitting analyses are shown in p = 0.91 for CP occurrence; p = 0.87 for CP intensity). Furthermore, no evidence emerged for interactions of these components with age.For both CP occurrence and intensity, within-pair MZ correlations were remarkably higher than DZ correlations, with the latter being of vanishingly low magnitude. A similar pattern was observed when stratifying the sample by zygosity and sex; however, a strong random variability prevented the easy interpretation of the estimates, in particular for a higher (yet not significantly higher) correlation in unlike-sex DZ twins compared with same-sex DZ twins. The correlation pattern did not suggest a role for shared  effects on trait liabilities, but did suggest a key role for non-additive (possibly epistatic) genetic influences. Accordingly, the ADE structural equation models  provided a better description of the data than the ACE models . Under these models, for both CP occurrence and intensity, a negligible additive genetic contribution was found in the sex-unstratified analysis, and in both males and females. Broad  heritability estimates were basically driven by non-additive genetic effects, with values in the two-group analysis of 0.36 (A = 0) and 0.31 (A = 0.02) for CP occurrence and intensity, respectively. No sex differences were detected in the genetic and environmental variance components  and to promote healthy lifestyles, including exercise, diet and nutrition, stress management, self-care, and appropriate health-seeking behaviors . SpecifiIt should also be emphasized that the genetic and environmental components of CP occurrence and intensity were found not to be moderated by sex or age. Therefore, given the increasing prevalence of CP with age, this finding draws attention to possible age-related differences in gene–environment interaction, namely a higher genetic responsiveness to environmental stressors and triggers in older than in younger adults. Furthermore, the invariant etiological architecture of CP across sex and age may provide the rationale for combining cohorts with different demographic characteristics, at least among the Italian population, in order to have a higher statistical power for hypothesis testing. More importantly, if population-specific effects could be excluded at least between countries with similar cultural contexts, then demographically heterogeneous samples could also be combined across twin registers from these countries to investigate CP-related hypotheses on very large cohorts of twins. To this end, information from the Italian twin sample is currently being integrated with data from the Murcia Twin Registry, obtained with a similar survey and with the same CP questionnaire, and possible etiological heterogeneity is being tested to evaluate the feasibility of combined analyses.A major weakness of this study is the sample size, which, though respectable, was limited by the categorical nature of the variables, and did not allow reliable testing for sex differences in genetic and environmental effects, or for age differences, by exploring these effects in different age groups. Furthermore, with this sample size, it was considered over-ambitious to apply a multivariate design to incorporate information on CP comorbidities such as depression symptomatology or sleep problems, which were also collected for the study subjects. This limitation could be overcome through the above-mentioned collaboration between the Italian and the Murcia Twin Registries. Another limitation concerns the ethnic background of the study sample, which was characterized by adult twins of Caucasian origin only. Therefore, the findings of our study cannot be directly extended to other ethnic groups.This is the first twin study exploring the etiological bases of CP as a wide phenotype and its intensity in a non-clinical population of both male and female subjects using a comprehensively validated self-report questionnaire. Our results show similar etiologies for CP occurrence and intensity, with moderate genetic influences and large individual-specific environmental effects, and no evidence for age- or sex-moderation. Genetic influences are likely to encompass epistatic gene–gene interactions that should be further unraveled and quantified. The environmental effects draw attention to prevention opportunities that have currently only partially been exploited.


Fusarium oxysporum f. sp. conglutinans (FOC), the causal agent of cabbage fusarium wilt, is a serious threat to cabbage production in northern China, and most Chinese FOC isolates were identified as FOC race 1 (FOC1). To better understand the genetic diversity of FOC1 in northern China, we collected FOC isolates from five provinces in northern China and identified them as FOC1 through pathogenicity and race test. To evaluate the genome-level diversity of FOC1, we performed a genome assembly for a FOC1 isolate (FoYQ-1) collected from Yanqing, Beijing, where cabbage fusarium wilt was first reported in China. Using resequencing data of FOC1 isolates, we conducted a genome-wide SNP (single nucleotide polymorphism) analysis to investigate the genetic diversity and population structure of FOC1 isolates in northern China. Our study indicated that Chinese FOC1 can be grouped into four populations and revealed that the genetic diversity of FOC1 were closely associated with geographical locations. Our study further suggests that genetic differentiation occurred when FOC1 spread to the northwest provinces from Beijing Province in China. The FOC1 genetic diversity based on whole-genome SNPs could deepen our understanding of FOC1 variation and provide clues for the control of cabbage fusarium wilt in China. Fusarium oxysporum (FO) is one of soil-inhabiting fungus causing vascular wilt or root rot in over 120 economically important crops worldwide [formae speciales (f. sp.), and some formae speciales of FO can be further divided into several physiological races [forma specialis of FO are often polyphyletic. Based on sequence similarity for EF-1a gene and ITS sequences, individuals from different formae speciales may be more closely related to each other than individuals in the same forma specialis, suggesting that pathogenicity to certain plant hosts has independently arisen multiple times from distinct lineages [The fungus lineages . This molineages .Fusarium oxysporum f. sp. conglutinans (FOC), the causal agent of cabbage Fusarium wilt, is a worldwide threat to cabbage production, resulting in severe economic losses [c losses . Based oc losses . FOC1 isc losses , while rc losses ,9. Cabbac losses . Especiac losses . In 2001c losses , and thec losses . Cabbagec losses .Some studies showed that with the spread of FOC1 in northern China, the pathogenicity of FOC1 is differentiated, which indicates that FOC1 is differentiated ,14. TherThus far, most of the genetic diversity research in FO is to investigate the distribution of different physiological races. Physiological races also differentiate during the process of pathogen spread . KnowledIn this study, we selected three regions in Beijing Province , two regions in Hebei Province (Baoding and Xintai), two regions in Shanxi Province (Jinzhong and Taiyuan), two regions in Shaanxi Province (Weinan and Xi’an), and two regions in Gansu Province (Dingxi and Yuzhong) for collecting the diseased cabbage, respectively . For eacThe morphological analyses of isolated strains were performed to check colony morphology, mycelia, and conidia characteristics. As shown in We further performed a PCR-based molecular detection on the isolates by using FOC-specific primers. Each of 26 FOC isolates can be amplified a 346 bp FOC-specific DNA fragment, which is the same as that of 52,557 and 58,385, and the 346 bp fragment was not detected in other five FO . As a rePathogenicity and a race test of FOC isolates were carried out on nine cabbage cultivars, including two differential hosts (“XiaQiang” and “Zhonggan18”), and standard FOC1  and FOC2  were used as control. As shown in Combined with the above results, we confirmed that all FOC isolates were the FOC1.We selected FoYQ-1, which is a FOC1 isolate from Yanqing, Beijing Province, where cabbage fusarium wilt was first reported in China, for genome sequencing. The FoYQ-1 genome size was estimated to be 67 Mb on the basis of the k-mer statistics  from a pA total of 18,319 protein-coding genes were predicted for FoYQ-1. Among the predicted genes, 15,202 (80.51%) genes were functionally annotated. BUSCO assessment of the FoYQ-1 genome showed that 1325 (92.05%) of the gene models were complete , suggestforma specialis strains were used to construct phylogenetic trees, respectively  and Fowe (host is melon) clustered with FOC1 isolated collected from Xingtai on ITS phylogenetic tree. Focub and Foco (host is cotton) are clustered with FOC1 isolated from Xi’an on EF-1α tree. It is not clear whether some other forma specialis of FO and FOC1 are genetically close, or phylogenetic trees constructed by ITS and EF-1a sequences cannot distinguish the genetic distance between them. The EF-1a phylogenetic trees showed that some FOC1 isolates from the same province were clustered together. For example, FoBD-1, FoBD-2, FoXT-1, and FoXT-2, collected from Hebei Province, were clustered together. However, clustering according to geographical regions was not found in the ITS phylogenetic tree. Our study suggested that EF-1α phylogenetic trees may better reflect the genetic relationship of FOC1 from different geographical regions compared with ITS phylogenetic trees.The ITS and the EF-1α sequence of all 26 FOC1 isolates showed the highest identities  with published FO strains sequences when blasted in the NCBI database . These Iectively a,b. BothThe genetic variation and population structure among the FOC1 in northern China were evaluated by resequencing the 26 isolates. We obtained a total of 31.2 Gb, with median depth of 17-fold and 99.21% coverage of the assembled FoYQ-1 genome. After aligning the reads to the FoYQ-1 genome, we identified a total of 264,218 SNPs, with an average of 4.24 SNPs per kb. A total of 13,578 SNPs (5.18%) were located in the coding regions, among which 7098 were non-synonymous SNPs. We used the SNPs to construct a population structure for FOC1 by admixture and explored the population structure of FOC using different delta K  values  a. As shoIn the control of fusarium wilt, resistant varieties are the most effective measures. Therefore, accurate identification of FO physiological races is essential for disease control . CabbageThe genetic diversity of FO has also been widely studied. The translation EF-1a and ITS sequences were used in these studies, and many reports showed that compared with ITS sequence, the EF-1a was a suitable genetic marker to distinguish between species. However, many studies have shown that EF-1a cannot fully reflect the genetic structure of FO ,18,19. Hforma specialis of FO. This chromosome determines the forma specialis type of FO. Transferring the special chromosome to different FO can change the forma specialis of FO. More and more studies have shown that the physiological race of FO is determined by effectors on the same special chromosome [Ma et al. found that the genome of FO is divided into core genome and accessory genome . The accromosome ,26,27. Promosome . TherefoIn conclusion, we provided genome data of native FOC1 and first used genome-wide SNPs to analyze the genetic diversity and population structure of FOC1 isolates collected from northern China. Our results revealed that the genetic groups of FOC1 were closely associated with geographical locations of pathogen collection and indicate that genetic differentiation occurred when FOC1 spread to the northwest provinces form Beijing Province in China. Our work contributed to the knowledge of FOC1 differentiation in northern China and can provide useful information for cabbage resistance breeding in China.The strains from Fujian of China were a friendly provision from Xiao Rong-feng of the Fujian Academy of Agriculture Science. The strains from Zhejiang of China were a friendly provision from Zhang Zheng-guang of the Zhejiang University. The single-spore cultures of FOC1 were isolated from diseased plants in Beijing, Gansu, Shanxi, Hebei, and Shaanxi Provinces cabbage fields .We selected 11 counties of Beijing, Hebei, Shanxi, Gansu, and Shaanxi Provinces in northern China as sampling points. In each county, diseased plants were collected from 2 to 3 diseased fields in different villages. As a result, a total of 26 sampling regions were selected. FOC isolates were collected from diseased plant tissues according to traditional protocol , and theA total of nine cabbage cultivars were used as differential hosts to identify races and pathogenicity of the FOC1 isolates. In nine cabbage cultivars, “Xiaqiang”, “Huifeng7”, and “Zhonggan18” are resistant (R) to FOC race 1 (FOC1) but susceptible (S) to FOC race 2 (FOC2). “Huifeng6”, “Lutailang”, “Zhonggan96”, and “Zhenqi” are R to FOC1 and R to FOC2. “Hanchun1” and “Zhonggan21” are S to FOC1 and S to FOC2.For single-molecule real-time (SMRT) sequencing, only subreads with length more than 500 bp and RQ value higher than 0.75 were retained for future analysis. Canu (v1.6)  assembleMolecular analysis was performed including the FOC-specific, ITS sequence, and EF-1a sequence. The primers for FOC-specific were 5′-TCAATGATAGTGACAAGGGTTT and 5′-AATTTGCTGTGATAGGTGGAT, which can amplify a FOC-specific DNA fragment with size of 346 bp . The priThe nucleotide sequences of the ITS region and EF-1a gene were aligned with ClustalW. Phylogenetic tree was generated by the neighbor-joining (NJ) method with default parameters from the alignment of the nucleotide sequences with MEGA10. Bootstrap analysis with 1000 replications was performed to assess group support.For resequencing, a total of 26 isolates were sequenced, and 5 ug DNA for each isolate was used for library construction. The libraries with 350 bp insert size were sequenced on the Illumina HiSeq 2500, generating 150 bp paired-end reads. Raw reads were subjected to removal of adaptors and trimmed of low-quality bases. We obtained a total of 31.2 Gb clean data, with median depth of 17× coverage for each isolate. The clean data of each individual were mapped to the FoYQ-1 genome using BWA (Version 0.6.2). Then, samtools  was used to transform the format and sort the result of BWA. The variations were called using GATK  according to the GenotypeGVCFs pipeline. Plink software with the parameters -geno 0.2, -maf 0.05 was used to filter the variations and obtain high-quality SNPs and small indels.The ADMIXTURE software was used to construct a population structure of FOC1 isolates (with the parameters –C 0.001 –j10 –seed1). A custom R script was used to confirm and plot the optimal population structure.


In the last few decades, a dramatic increase in the global prevalence of allergic diseases and asthma was observed. It was hypothesized that diet may be an important immunomodulatory factor influencing susceptibility to allergic diseases. Fermented food, a natural source of living microorganisms and bioactive compounds, has been demonstrated to possess health-promoting potentials and seems to be a promising strategy to reduce the risk of various immune-related diseases, such as allergic diseases and asthma. The exact mechanisms by which allergic diseases and asthma can be alleviated or prevented by fermented food are not well understood; however, its potential to exert an effect through modulating the immune response and influencing the gut microbiota has been recently studied. In this review, we provide the current knowledge on the role of diet, including fermented foods, in preventing or treating allergic diseases and asthma. The prevalence of allergic diseases and asthma has increased significantly in the last decades, especially in the western world . This phPresently in the western world, we observe the fashion for including natural ingredients in the diet, which increases the popularity of fermented food. However, fermented food has been a part of the human diet for centuries. In the past, fermentation was a method of natural preservation of meat, fish, vegetables, fruits, and dairy products. The benefits of a diet containing fermented milk were first described by the Nobel Prize winning microbiologist Ilya Metchnikoff over a century ago. He noticed that people who regularly consumed fermented yogurts regularly lived longer, and proposed that this may be connected with the beneficial action of lactobacilli . In receIn this paper, we present the current knowledge on the role of fermented food in asthma and respiratory allergies, with special attention given to the possibilities of their application as a potential primary prevention mechanism for these diseases. We present the latest experimental and clinical research and discuss the mechanisms of action of different types of fermented food in allergic diseases and asthma. Treating and preventing asthma and respiratory allergies with a diet containing fermented products appears to be a promising strategy.Lactobacillus, Lactococcus, and Tetragenococcus, are widely present in many fermented foods, especially fermented milk and soy products [Staphylococcus, Enterococcus, Brevibacterium, and Propionibacterium [Bacillus are present in legume-based fermented foods, such as natto, miso, and soy sauce, made from soybeans [Saccharomyces) involved in the fermentation of wine and beer bread [Penicillium, Aspergillus, and Rhizopus, are widely associated with dairy, meat, and soy product fermentation [Fermented foods are generally defined as foods and beverages created through desired microbial growth and enzymatic conversion of food components . The widproducts ,12. Aparacterium ,10,13. Ssoybeans ,10. Amoner bread . Differeentation ,10,11,13Microorganisms specify the type and outcomes of fermentation processes that can be categorized as alcoholic and anaerobic lactic acid fermentations, which occur in yoghurt, kimchi, and wine, or aerobic fermentations used by fungi in the processing of soy sauce and miso and by AAB in the production of vinegar and kombucha ,11,14.Fermentation can also be classified on the basis of the primary source of fermentative microorganisms ,8. NaturOf note, certain fermented products  are commonly rid of the fermentation microorganisms before being distributed and may not comprise living microorganisms at the consuming stage. They should be categorized as “foods made by fermentation” ,9. LastlFermentation is an effective method of food processing which provides many valuable qualities, namely extended shelf-life, improvement of safety, attractive flavor and texture, enhanced nutritional and functional properties, and health benefits ,8,17. Ep6 and 109 microbial cells per gram [Primarily, fermented foods have been found to contain living, functional microorganisms in notable quantities, ranging between at least 10per gram ,22. Studper gram ,22. Funcper gram ,23.Lactobacillus spp. and S. thermophiles, as it has been shown that cultured dairy products consistently contained the highest levels of LAB [Lactobacillus plantarum and Lactobacillus rhamnosus are also incorporated in oat-based fermented beverages that also contain the prebiotic fiber β–glucan [Lactobacillus plantarum isolated from sauerkraut and kimchi [Lactobacillus fermentum) classified as probiotics [10 microbial cells, which is suggested to be necessary to change the gut microbiota and supply potential health benefits [Many species present in fermented foods are genetically similar to or share physiological traits with strains used as probiotics; hence, the health benefits of fermented foods may be due to probiotic activity ,7,16. Yos of LAB ,16,20,24β–glucan . Probiotd kimchi ,26,27,28obiotics . It can obiotics ,31. Intebenefits .Additionally, microbial activity during food fermentation results in the improvement of food safety and nutritional composition. Numerous species of LAB derived from fermented vegetables and milk products produce antimicrobial end-products, such as organic acids, ethanol, bacteriocin, and nisin, and thereby diminish the risk of pathogen contamination ,23,24,34Fermentation-related microorganisms have the ability to synthesize numerous metabolites and enhance the food with new nutrients and bioactive molecules that can exhibit either beneficial or harmful properties to human health . In partFinally, according to observational and interventional studies, fermented foods can also modulate the gastrointestinal microbiota composition or function and redress the dysbiosis ,60. AmonBased on the knowledge of health-promoting components in fermented food and their potential ability to interplay with the immune system and the intestinal microbiota, the intake of fermented foods seems to be a promising strategy to reduce the risk of several disorders, including immune-related diseases, such as allergic diseases and asthma.The exact mechanisms by which allergic diseases and asthma might be reduced or prevented by consuming fermented foods are uncertain. The most widely accepted theories are that fermented foods lead to anti-allergic effects either by modifying/influencing the gut microbiota or systemically modulating the immune response. Additionally, fermentation is an efficient approach for reducing or eliminating food allergenicity.Immunomodulation applies to the concept that fermented foods can change the activity of both the local and systemic immune system, leading to upregulation or downregulation of the response to a successive challenge . The immFirstly, fermented food components might modify mucosal immunity and restore or enhance the barrier function of epithelium, either in the gut or lung. Indeed, fermentation-derived metabolites could increase the expression of tight junction proteins between epithelial cells and regulate the mitogen-activated protein kinases (MAPK) signaling pathway, thereby inhibiting the permeability of allergens via paracellular diffusion route ,65,66,67Bifidobacterium, Lactobacillus, and Akkermansia, and altering the ratio of Firmicutes to Bacteroides [Diet is an important modifiable factor influencing the gut microbiota . Fermentteroides ,59,60,92teroides ,94,95,96teroides . The preteroides ,95. Anotteroides ,94. SCFAteroides ,96,98. Iteroides . Based oTo date, the various types of fermented foods have been extensively studied using in vitro experiments or animal models to support their immunomodulatory and anti-allergic effects. Clinical studies on fermented foods have also been conducted, although there are considerably fewer.Lactobacillus bulgaricus, Streptococcus thermophilus, and Lactobacillus paracasei strains was observed to mitigate respiratory allergy in ovoalbumin (OVA)-sensitised BALB/c adult mice [Fermented dairy products such as yogurt, kefir, and cheese have received special attention because these products remain the main source of living microorganisms, mainly LAB, that are capable of modifying the immune response ,16,20,24ult mice . The maiult mice . More reult mice .Lb. kefiranofaciens and Lb. kefiri have also been shown to exhibit anti-allergenic effects regarding Th1/Th2 balance in the favor of Th1 cells by induction of IFN-γ, TNF-α, IL-6, IL-12, and IL-1β in murine peritoneal macrophages [Lb. kefiranofaciens M1 isolated from kefir grains was found to reduce the levels of IL-5, while increasing the levels of IL-12, elevating the percentage of CD4+CD25+ Tregs, and inhibiting the IgE production in OVA-sensitized Th2-polarized mice [Lb. kefiranofaciens can prevent the development of allergen-induced sensitization by an increase of Treg activity, a suppression of Th2  and Th17 cytokines in splenocytes and BAL fluid, and an inhibition of specific IgE production in serum. It was also clearly demonstrated that Lb. kefiranofaciens is able to suppress the features of the asthmatic phenotype, including AHR to methacholine, airway inflammation, eosinophil infiltration in the lung, the hypersecretion of mucus, and goblet and smooth muscle cell hyperplasia in OVA-allergic asthma mice [Lb. kefiranofaciens, have also been shown to modulate the gut microbiota composition by stimulating the growth of Lactobacillus, Lactococcus, and Bifidobacterium and decreasing Proteobacteria and Enterobacteriaceae abundance, as revealed in several animal studies. Moreover, kefir consumption altered the Firmicutes/Bacteroidetes ratio which has been associated with maintaining homeostasis [The kefir-derived LAB strains rophages ,72. Addihma mice . Kefir aeostasis ,101,102.Lactobacillus helveticus isolated from fermented milk and some types of hard or semi-hard cheese were selected as an immunomodulatory effect strain. Experimental study in a murine pollen allergy model indicated that Lactobacillus helveticus LH2171 significantly diminished antigen-induced and total IgE levels in the blood. Additionally, Lactobacillus helveticus LH2171 inhibited the proliferation of immune cells and cytokine secretion in the submandibular lymph node and upregulated the expression of IL-10 and Foxp3, a marker of Tregs, in Peyer’s patches [Lactobacillus helveticus LH2171 using the antigen-stimulated murine naïve splenocytes in vitro and demonstrated the upregulation of IFN-γ and IL-10 secretion, with the suppressive effect of this strain on IL-4 and IL-13 production. Surprisingly, the impact on Treg cells differentiation as well as inhibition of systemic anaphylactic reaction was not observed. Further analysis using a murine model for pollen allergy revealed that Lactobacillus helveticus LH2171 suppresses pollen allergy symptoms, including face scratching and sneezing frequency. However, the alleviation of symptoms was not accompanied by inhibition of antigen-specific IgE production [Recently,  patches . Makino oduction .Lactobacillus helveticus LH2171 strain for 12 weeks significantly improved ocular and nasal symptoms, including the number of sneezes. However, no significant differences were found in concentrations of Th1 and Th2 cytokine and total IgE or antigen-specific IgE levels in the serum [Lactobacillus helveticus LH2171 for 16 weeks improved nasal discomfort and congestion in subjects with perennial allergy. The nasal and blood eosinophil counts and the eosinophil dynamics were markedly decreased in the Lactobacillus helveticus LH2171 group compared to the placebo group, suggesting that the lower degree of “stuffy nose” in this group was possibly due to suppression of eosinophils [A randomized, double-blind, placebo-controlled study conducted in adult patients with mite or house dust allergy indicated that the daily consumption of drinkable yogurt containing he serum . More reinophils .Lactobacillus species on asthma via gut microbiome modulation has been investigated in the murine house dust mite (HDM)-induced model of asthma. These findings indicated that Lb. reuteri isolated from fermented milk was the most effective, and modulated gut microbiota by increasing the abundance of Lactobacillus, Bifidobacterium, and Enterococcus. L. reuteri altered the gut microbial function toward increased SCFA, mainly butyrate production, and in turn decreased total IgE, inhibited Th2-associated proinflammatory cytokines , and reduced airway inflammation [Lactobacillus casei strains, broadly used in fermented dairy products, exhibited preventive effects on HDM-induced asthma in mice by modulating both the immune response and the gut microbiota composition. Lb. casei strains decreased the number of lymphocytes, eosinophils, and neutrophils and the levels of Th2 cytokines  and Th17 cytokine (IL-17A) as well as chemokine (such as eotaxin-1) in BALF. In addition, Lb. casei strains inhibited both HDM-specific IgG1 and total IgE levels in the serum. However, the results are not consistent for all strains, as it has been shown that L. casei1 did not affect the production of inflammatory cytokines and L. casei1 has no impact on immunoglobulin levels, while L. casei3 exhibited the highest preventive effect. Furthermore, all five L. casei strains improved the diversity of gut microbiota mostly by promoting the growth of Firmicutes and increasing the SCFA (acetate and propionate) contents [Lactobacillus species, especially L. reuteri and L. casei3 supplementation, may become an effective approach to prevent asthma and other allergic diseases.More recently, the preventive effect of six ammation . A furthcontents . Taken tDairy products are a rich source of conjugated linoleic acids (CLA), a class of unsaturated fatty acids, and their content can be significantly enriched by LAB fermentation ,57. In tFermentation of different plants leads to the synthesis of various bioactive compounds that are not present in raw products, including dietary fiber metabolites, phospholipids, isoflavones, phenolic acids, saponins, and phytic acids . The immCordyceps militaris fermented with a Pediococcus pentosaceus strain demonstrated anti-allergic activity by suppressing the release of proinflammatory cytokines, inhibition of the signal transduction cascade accountable for type I hypersensitive allergic reaction in IgE/Ag-activated RBL-2H3 cells, and decreasing infiltration of immune cells and skin swelling in the in vivo BALB/c mice model [Ramulus mori, Salvia plebeia, and Anthriscus sylvestris in the experimental study using cells from the human lung epithelium in vitro and an OVA/LPS-induced asthma mouse model in vivo. The administration of this extract significantly inhibited the secretion of proinflammatory cytokines , which in turn decreased IgE level and inflammatory cell infiltration in the BALF and hence alleviated lung tissue inflammation [Asparagus cochinchinensis has been found to exhibit the dose-depended anti-asthmatic effects expressed through decreasing specific IgE level as well as reduction of immune cells infiltration, secretion of mucus, and respiratory epithelium remodeling in vivo in the OVA-challenged asthma mice model [Platycodon grandiflorum root extracts containing triterpenoid saponin platycodin D inhibited IL-17E cytokine production, and in turn the numbers of eosinophils and total cells BALF obtained from LPS/OVA-induced asthma mice, which indicates their effect on reducing airway inflammation [Experimental studies provide growing evidence for the anti-allergic and anti-asthmatic activity of fermented products based on herbal components or extracts derived from medicinal plants or fungi. For example, well-known medicinal fungus ce model . These ece model . The antammation . The antammation ,48. Sapoammation ,23. Indece model . Recentlammation .Panax spp., has been used around the world as the functional food and herbal medicine for treatment of allergic diseases including asthma and allergic rhinitis. Scientific research strongly supports the anti-allergic properties of ginseng extracts, which are additionally enforced by the fermentation [Panax ginseng extract reinforced its anti-allergic effects in vitro and in vivo. Ginseng fermented with Lactobacillus plantarum more strongly inhibited secretion of FcεRI-mediated β-hexosaminidase and IL-4 in mast cells and IgE-mediated passive cutaneous anaphylaxis in mice compared with unfermented extracts [Ginseng, a dried root of the entation . It was extracts . Lee et extracts . Furtherextracts . Less coextracts . In addiextracts . Ginsenoextracts .Double-blind, placebo-controlled study of the safety and efficacy of fermented red ginseng has confirmed the significant improvement in both nasal congestions symptoms and overall quality of life after a 4-week medication in patients with persistent allergic rhinitis. In addition, skin reactivity to perennial allergens was markedly decreased in treated patients, suggesting that fermented red ginseng can effectively suppress allergic inflammatory reactions .Lactococcus lactis A17 isolated from fermented cabbage in OVA-sensitized BALB/c mice in vivo model and was related to the modulation of TLR4 expression and subsequent suppression of IL-4 cytokine and specific IgE production [Lactobacillus plantarum extracted from traditional Chinese sauerkraut Paocai inhibited allergen-induced infiltration of the inflammatory cells in the lung tissues of BALB/c mice in vivo via the Tregs-dependent mechanism. All tested strains induced the Th1-associated cytokines; however, only one strain (Lp) has been shown to be effective in inhibiting the Th2 cytokine IL-4 production, suggesting the strain-dependent effect. Surprisingly, the expected effects on the levels of specific IgE were not found [Lactobacillus plantarum K-1 isolated from kimchi can mitigate allergic diseases, such as anaphylaxis, atopic dermatitis, and rhinitis, by suppressing the expression of TNF-α and IgE-switching cytokine IL-4 via inhibition of NF-κB and AP-1 signaling pathways [Lactobacillus sakei, another probiotic strain extracted from kimchi, was observed to restore the skin epidermis, diminish the skin lesions, and reduce IgE level in in vivo mouse models of DNCB-induced atopic dermatitis. Oral administration of this strain decreased the number of CD4+CD25+ T cells and B cells and suppressed the Th2-assosiated cytokines by in vitro stimulation of tolerogenic DCs and further promotion of Treg differentiation [Lactobacillus sakei modulated the structure of gut microbiota, decreasing AD-related gut bacteria  abundance while increasing the relative abundance of Ruminococcus that might be associated with Treg activation, and significantly reduced symptoms in atopic dermatitis mice [Sauerkraut, one of the most popular and oldest types of fermented cabbage, was considered a functional food with an immunomodulatory activity, as it is a unique source of vitamins, phenolic compounds, and in particular, LAB ,67. The oduction . Moreoveot found . Kimchi,pathways . Lactobantiation ,69. In atis mice .Pediococcus pentosaceus exhibit a suppressive impact on IgE/Ag-mediated type I hypersensitivity reactions through inhibition of mast cell degranulation, reduction of proinflammatory cytokine expression, and negative regulation of the FcεRI-mediated signaling pathway [LAB-fermented germinated brown rice is another plant food which looks promising in terms of the treatment of IgE-dependent allergic diseases, as it contains a high level of ferulic acid and protocatechuic acid, both known to have anti-allergic activities . In addi pathway .Saccharomyces cerevisiae. In the in vivo study model, oral intake of the fermented multi-fruit beverage was demonstrated to change the Th1/Th2 balance by enhancement of Th1-type immune response with subsequent attenuation of Th2-related cytokine IL-4 and IL-5 secretion in OVA-sensitized BALB/c mice [Lactobacillus casei-fermented blueberry pomace revealed potential gut-improving functions through suppressing the growth of Escherichia coli and Enterococcus and altering the ratio of Firmicutes and Bacteroidetes as well as the stimulation of Bifidobacterium, Ruminococcus, Lactobacillus, and Akkermansia genera. In addition, fermented blueberries increased the production of SCFA, especially acetic, butyric, and lactic acids [L.casei-fermented blueberry pomace [Fermented fruits are a rich source of diverse micronutrients, dietary fiber, and phytochemicals such as anthocyanins, flavanols, resveratrol, and phenolic acids that have been found to present multiple health-promoting benefits ,67. A reB/c mice . The immB/c mice . BerriesB/c mice . EvidencB/c mice . Berry pB/c mice ,67. Receic acids . These fy pomace .Lactobacillus plantarum YIT 0132 against allergic symptoms in an animal model of Japanese cedar pollinosis, which is the most common type of allergic rhinitis in Japan. In addition, the daily intake of citrus juice fermented with Lactobacillus plantarum alleviates symptoms and enhances quality of life in patients with Japanese cedar pollinosis during the natural pollen season [L.plantarum-fermented juice. Furthermore, these benefits were associated with the immunomodulating activities of Lactobacillus plantarum, as the reduction of Th2 cells, total IgE, and eosinophil cationic protein level was observed [Harima-Mizusawa et al. demonstrated the beneficial effect of citrus juice fermented with n season . Furtherobserved .Aspergillus oryzae and lactic acid bacteria (Pediococcus parvulus and Enterococcus faecium), is an excellent source of isoflavones and has been shown to suppress peanut hypersensitivity on a murine model of peanut allergy as evidenced by a reduction of Th2 cytokines  and peanut-specific IgE production [Numerous fermented soybean foods from different parts of Asia, namely tempeh, natto, miso, douche, and soy sauce, have long been consumed due to their well-known health benefits ,119. Evioduction . Recentloduction . The autoduction .Lactobacillus spp., Bacillus strains (B. subtilis or B. licheniformis), Enterococcus faecium, and Leuconostoc spp. [Bacteroidetes, Clostridium leptum, and Bacteroides fragilis and altered or decreased the ratio of Firmicutes/Bacteroidetes [Bifidobacterium, Lactobacillus, Escherichia coli, and Enterococcus species [Cheonggukjang (fermented soybean paste), a traditional Korean fermented dish also called natto, tempeh, and douchi in other Asian regions, has recently emerged as a functional food with immunomodulation properties . Cheonggtoc spp. . The anttoc spp. . Later stoc spp. . Additiotoc spp. . These etoc spp. . Tempeh,roidetes . Moreove species .Questionnaire-based studies have reported that ingestion of foods with increased content of soy genistein might be correlated with better clinical control of asthma symptoms and improvement of lung function in adult patients ,125. HowRecently, the pea (Pisum sativum), belonging to legumes, has received special attention, as it offers an opportunity to replace animal protein with a healthier, economically viable, and environmentally friendly source of protein. Pea seeds contain high amounts of essential amino acids and proteins, dietary fiber, minerals, vitamins, and phytochemicals that are related to their health beneficial properties . ExperimWhen summarizing the evidence from the studies reviewed here, it seems rational to conclude that the consumption of fermented foods can play a promising role in treatment or even prevention of asthma and respiratory allergies. The vast majority of published studies provide data supporting the positive anti-inflammatory, anti-allergic, and immunomodulatory potential of different fermented foods ,61,62,67Evidence from epidemiological studies supports the hypothesis that dietary habits affecting the intestinal microflora may contribute to reducing the risk for developing allergies. One example is an anthroposophic lifestyle, characterized by high intake of fermented vegetables and biodynamic food, restrictive use of antibiotics, incomplete immunization, and prolonged breastfeeding . In one Data from the large Korea National Health and Nutrition Examination Survey (KNHANES) study conducted to assess the association between dietary factors and asthma in adults demonstrated a significant inverse correlation between kimchi consumption and the prevalence of adult asthma. It has been observed that patients with asthma reported a lower consumption of kimchi and fish compared to those without asthma . In the When summarizing the results from epidemiological studies, it is important to highlight the potential limitation of this research method. Questionnaire-based assessment of exposure creates the possibility of recall or selection bias, as well as the risk of misunderstanding the question about the type of exposure. Furthermore, the correlations between outcomes and exposure found in the observational studies, even if biologically plausible, do not necessarily imply a causal relationship and require further analysis before making detailed conclusions.Fermented foods have been considered as an essential element of the human diet for thousands of years, enriching it with nutritional compounds. Presently, the consumption of fermented foods is gaining increasing interest as an important dietary method for improving human health. Their excellent biological activities, in particular anti-inflammatory and immunomodulatory properties, are associated with high content of live microorganisms, phytochemicals, and bioactive compounds. Following the growing evidence from epidemiological studies, the Metchnikoff’s original theory can be extrapolated to the possibility that fermented foods could represent a promising strategy of treatment and prevention of asthma and respiratory allergies. As reviewed, evidence from in vitro experiments and animal trials reported in the literature thus far has shown that fermented foods could exert effects against allergic airway inflammation, possibly through modulating the local and systemic immune response and modifying the gut microbiota. Although the concept of the food–gut axis, and further, the gut–lung axis has been the subject of intense study in recent years, detailed mechanistic studies are still required to obtain insight on the association between diet, in particular fermented foods, and gut and lung microbiota composition and their impact on the immune response. Furthermore, most of our knowledge in this field is based solely on experimental studies, while there are only limited clinical evidence. Therefore, in the future, there is a critical need for preventive and treatment-oriented, well-designed, randomized, controlled clinical trials and broad population studies to provide support to the evidence found on the effectiveness of fermented food on asthma and respiratory allergies. These studies should focus on well-defined disease phenotypes, diverse immune-related parameters, the gut–lung microbiome composition, and analysis by metagenomics and metabolomics. However, designing and repeating studies on fermented foods may be a challenge due to the high variability of microbial cultures and ingredients present even in the same types of fermented foods. Nevertheless, it has been suggested that this effort is important, as the benefits of whole fermented foods are bigger than the sum of their individual components. Such efforts are particularly needed, as diet is one of the most modifiable and readily available interventions for every person and may represent a safe, non-invasive, non-pharmacological addition to disease management. Meanwhile, fermented foods remain a promising strategy for the treatment and prevention of asthma and respiratory allergies that could be included into dietary guidelines in the future.


Gasterosteus aculeatus) to a rare derived habitat—nutrient‐depleted acidic lakes—based on whole‐genome sequence data. Sequencing marine stickleback from six locations across the Atlantic Ocean then allows us to infer that the frequency of these derived variants in the ancestral habitat is unrelated to the likely opportunity for gene flow of these variants from acidic‐adapted populations. This result is consistent with the selective neutrality of derived variants within the ancestor. Our study thus supports an underappreciated explanation for the maintenance of standing genetic variation, and calls for a better understanding of the fitness consequences of adaptive variation across habitats and genomic backgrounds.Adaptation to derived habitats often occurs from standing genetic variation. The maintenance within ancestral populations of genetic variants favourable in derived habitats is commonly ascribed to long‐term antagonism between purifying selection and gene flow resulting from hybridization across habitats. A largely unexplored alternative idea based on quantitative genetic models of polygenic adaptation is that variants favoured in derived habitats are neutral in ancestral populations when their frequency is relatively low. To explore the latter, we first identify genetic variants important to the adaptation of threespine stickleback fish ( Adaptation to acidic waters probably involves numerous traits, but particularly obvious elements include the reduction of external skeletal armour and body size in some acid‐adapted stickleback populations relative to their ancestor (and to standard freshwater‐adapted stickleback)  Figure . The fun) Figure  . An exc) Figure , an isla) Figure , the eas) Figure . This pa) Figure . What reTo address this question, we here use whole‐genome sequence data to examine SGV in marine stickleback across the Atlantic Ocean. We hypothesize that if the presence of SGV relevant to acidic adaptation in marine stickleback around North Uist reflects a balance between gene flow and purifying selection, the frequency of alleles favoured in acidic habitats should be elevated in marine stickleback breeding around North Uist compared to marine stickleback sampled from more distant locations Figure , top. Th22.1Gasterosteus aculeatus. We chose 20 individuals from each of these freshwater populations at random and combined their DNA to equal molarity without PCR (polymerase chain reaction)‐enrichment into either an acidic or a basic pool of 100 individuals each. The goal of this pooling  was to obtain relatively precise allele frequency estimates for acidic versus basic stickleback in general, while ignoring allele frequencies within each specific population. To nevertheless have access to individual genotypes and haplotype information, we additionally chose two individuals from each acidic and basic population at random for individual sequencing (indSeq).A precondition for our analysis of SGV in marine stickleback was the initial identification of genetic polymorphisms important to acidic adaptation. For this, we considered five acidic and five basic lakes from North Uist from which individual DNA was already available , Ireland (IR), The Netherlands (NL), Germany (DE), Iceland (IS) and Eastern Canada (CA) Figure ,2019b, iThe 47 total DNA libraries  were paired‐end sequenced to 150 bp together on a single S4 flow cell of an Illumina NovaSeq 6000 instrument, producing a genome‐wide median read depth per base pair of 85× on average across the pools, and of 16× across the individuals . From the alignments, we derived nucleotide counts (pileups) for all genome‐wide positions by using the pileup function from the Rsamtoolsrpackage , constructed maximum‐likelihood genealogies, and visualized them as unrooted phylograms. Node support was determined based on 500 bootstrap iterations. As an alternative to phylograms, we also considered exploring population structure by ordination . However, the proportion of variation captured by the first ordination axes was consistently small (~8% or less). We therefore considered ordination an ineffective tool for pattern recognition.Based on the genotype files, genealogies were generated by using the 2.4AFD. Positions qualified as high‐differentiation SNPs if they showed AFD ≥ 0.85, approximately corresponding to the top 0.01 percent of the AFD distribution. This AFD threshold was more stringent than in Haenel et al. (AFD within 0.1% of the genome‐wide median) between the acidic and the basic pool. The latter criterion ensured that these SNPs did not tag genome regions (consistently) involved in acidic adaptation. At each of the adaptive SNPs, we then defined the nucleotide predominant in the acidic pool as the “acidic allele,” and determined and graphed the frequency of these alleles in all six marine sample pools. An analogous analysis was performed for the baseline SNPs, here defining the acidic allele as the one relatively more common in the acidic than the basic pool. Our prediction was that if genetic variation at the adaptive SNPs in marine stickleback reflects gene flow–selection balance, the frequency of the acidic alleles at these markers (but not at the baseline SNPs) should be elevated in marine stickleback sampled on North Uist. As a resource, we additionally compiled all genes located within a 100‐kb window centred at each adaptive SNP.To identify alleles important to the adaptation of stickleback to acidic habitats, we performed genome‐wide differentiation mapping between the acidic and basic sample pools. That is, we scanned the poolSeq SNPs for positions exhibiting extremely high global differentiation between stickleback from acidic vs. basic lakes. The reason why we did not define genetic variation important for acidic adaptation simply as SNPs highly differentiated between acidic and marine fish is that this would mostly have uncovered genetic variation important to marine–freshwater divergence in general. Such variation is abundant in North Uist stickleback Figure . Our focl et al.  (0.70) bl et al. . With thAFD = 0.96), the adaptive SNP tagging the genome region showing the strongest acidic–basic differentiation in a previous investigation .   Figure . These a) Figure , based o) Figure . However) Figure . Haploty) Figure . Across At the adaptive SNPs, marine stickleback generally exhibited lower frequencies for the alleles characteristic of acidic fish  than for the alleles typical of the basic populations (median frequency 0.70) Figure . Also, tall marine samples (n = 21; indicated in Table Overall, these findings are in line with observations in Haenel et al.  and indiThe finding of similar frequencies of alleles important to adaptation to acidic waters across Atlantic marine stickleback challenges perpetual antagonism between gene flow and purifying selection (Bassham et al., An important caveat to consider is that although acidic habitats and the associated stickleback ecomorphs Figure  are exce4Adaptation commonly occurs from standing genetic variation, but how this variation is maintained in ancestral populations is little explored. We have here presented observational evidence suggesting that, overall, genetic variants important to adaptation to a highly derived habitat are maintained at moderate frequencies within the ancestral habitat. These variants do not appear to occur in higher frequencies in geographical regions where ancestral populations have a higher opportunity for gene flow from derived populations. We thus conclude that long‐term gene flow–selection balance is an incomplete explanation for the maintenance of SGV. Instead, we propose that purifying selection of these variants in the ancestral habitat subsides as their frequency decreases, thus allowing their neutral persistence. This novel perspective on the maintenance of SGV should now be scrutinized by controlled experimental work quantifying the fitness consequences of individual genetic variants across different habitats and genomic backgrounds.The authors declare no competing interests.D.B. and Q.H. conceived the study; A.M. provided all freshwater and marine samples from North Uist; Q.H. performed wet laboratory work; Q.H., D.B. and L.G. wrote code and analysed genomic data; Q.H. and D.B. interpreted the results and wrote the manuscript.Supplementary MaterialClick here for additional data file.Supplementary MaterialClick here for additional data file.


Camellia oleifera germplasm resources were collected from Guizhou Province, but the fruit morphological variation and genetic diversity of C. oleifera germplasm resources remain unclear. The genetic diversity of C. oleifera germplasms resources in Guizhou was studied based on fruit traits and simple sequence repeat (SSR) molecular markers to build a core collection. This paper aims to provide a scientific basis for the collection, management, development, and utilization of C. oleifera resources in Guizhou province. The variation coefficients among and within varieties of seven fruit phenotypic traits of C. oleifera ranged from 11.79% to 61.76% and from 8.15% to 42.31%, respectively, showing rich phenotypic variation. Furthermore, 12 SSR markers were used to analyze the genetic diversity. These primers generated 214 polymorphic bands, and the average number was 17.833. The average number of effective alleles (Ne), Shannon’s information index (I), observed heterozygosity (Ho), expected heterozygosity (He), polymorphic information content (PIC), and major allele frequency (MAF) were 8.999, 2.301, 0.965, 0.50, 0.836, and 0.238, respectively. The results showed that 12 SSR markers had high polymorphism, and the genetic diversity of 167 C. oleifera germplasm resources was high. Based on SSR molecular marker information and fruit traits clustering, 167 C. oleifera germplasm resources were divided into three groups. When constructing core collections based on fruit traits and molecular marker information, the PowerCore-25 of core collections greatly preserves fruit traits and improves genetic diversity. This paper can provide a reference for the genetic diversity and fruit traits variation of C. camellia germplasm resources in Guizhou Province. It is significant for establishing a core collection, thus promoting germplasm innovation and the development of the oil tea industry in Guizhou.Many  C. oleifera, belongs to the genus Camellia of the Teaceae family [C. camellia, olive (Olea europaea), oil palm (Elaeis guineensis Jacq.), and coconut (Cocos nucifera L.) are four important woody oil plants around the world [C. oleifera has high genetic diversity and phenotypic variation, and the study of genetic diversity is the basis of breeding better varieties. Therefore, understanding the genetic diversity of C. oleifolia and constructing a core germplasm resources bank of C. oleifolia is an important link in the development of local C. oleifolia.e family . C. camehe world . C. oleiPyrus [Juglans mandshurica Maxim [Cucumis melo L.) [Cucurbita spp.) [Punica granatum L.) [Catalpa fargesii f.duclouxii) [Ziziphus mauritiana Lam. [Phaseolus lunatus [Eucalyptus cloeziana F. Muell.) [Corylus avellana L.) [Akebia trifoliata (Thunb.) Koidz [Prunus sibirica [Pinus yunnanensis Franch. [Perilla [Cymbopogon winterianus Jowitt [Genetic diversity is the basis of biological diversity and is the driving force for the stability and continuous evolution of species. The methods for studying genetic diversity include morphology, cytology, and molecular marker technology. Morphology, as the most simple and convenient labeling method, can reveal the degree of genetic variation to a certain extent and explore potential target traits . With thPyrus , Juglansca Maxim , Tunisiamelo L.) , and pumta spp.) . It is ata spp.)  studied atum L.) , Huangxiclouxii) , and Zizana Lam. . Based o lunatus , Myrtace Muell.) , hazelnulana L.) , Akebia .) Koidz , and Prusibirica  were con Franch. , Perilla[Perilla , and Cyms Jowitt , the divC. oleifolia and evaluate the genetic diversity and relationships of C. oleifera germplasm resources in Guizhou Province. Constructing core collections of fruit traits and molecular marker information lays the foundation for the breeding of C. oleifolia and the protection of germplasm resources.This paper aims to establish the variation in fruit traits of The experimental samples were collected from key oil tea distribution areas in Guizhou Province from 2008 to 2011. Information about the collection area is shown in 1). After extraction, it was baked in an oven at 105 °C for 1 h to weigh the second mass (m2).The index of phenotypic and economic traits was determined using the method described by Yang et al. . In the The SSR molecular marker test includes DNA extraction, primer screening, fluorescence quantitative PCR, and electrophoresis tube detection. DNA was extracted using a genomic kit from Beijing Botanical Biofet Plant. This DNA extraction kit was used to select primers as the 36 main reference pairs ,22,23, aThe fluorescence quantitative PCR consists of a 20 µL system, including 17 µL of gold mix (green), 1 µL of front primers, 1 µL of back primers, and 1 µL of DNA template. The amplification procedures are listed as follows. First, pre-denaturation was performed at 98 °C for 2 min. The second stage was the cycle stage. Samples were denatured at 98 °C for 10 s, annealing at 55–62 °C for 10 s, extended at 72 °C for 10 s, and cycled 35 times. The third stage was the extension stage. Samples were extended at 72 °C for 5 min. For the capillary test, the mixing plate was heated at 95 °C for 5 min with a metal bath heater and immediately put into an ice box at −20 °C. The mixing plate was removed after cooling, centrifuged at 4000 rpm, thawed, and mixed well. Then, it was placed in an ABI 3730xl sequencer for capillary electrophoresis.t-test.The core collection was constructed with different sampling ratios . QGA software adopts Euclidean distance, multiple clustering priority sampling methods, and the shortest clustering method. PowerCore is an M strategy with a heuristic search for establishing core sets . The corThe exact loci and statistic genotyping data were analyzed with Gene Mapper 4.1. The genetic diversity indexes of SSR loci, including the number of different alleles (Na), number of effective alleles (Ne), Shannon’s information index (I), observed heterozygosity (Ho), expected heterozygosity (He), polymorphism information content (PIC), and major allele frequency (MAF) were calculated using GenAlEx 6.51 and PowerMarker software. Genetic structure and phylogenetic trees were constructed by means of Structure 2.3.4 and MEGA v.6. The quantitative traits were divided into 10 levels according to mean value (X) and standard deviation (σ), and Xi < X − 2σ and Xi > X + 2σ increased from 1 to 10 . Other dC. oleifera germplasm resources in Guizhou province. The F-value test showed that the fruit phenotypic traits exhibited significant differences within and among varieties . The highest variation within varieties was SGN, and the lowest was in FSI. The variation ranges of variance components among different varieties and within the varieties were 0.006–15.108 and 0–2.294, respectively. The trait with the largest variance component among different varieties was PT (67.35%), and the one with the smallest was SGN (43.68%). The phenotypic differentiation coefficients ranged from 80.15% to 95.56%, and the phenotypic differentiation coefficients of fruit traits were greater than 70%. The results indicate that the fruit traits mainly vary among different varieties, which is consistent with the results of the coefficient of variation  ranged from 0.592 to 0.927, with an average value of 0.836. The MAF ranged from 0.123 to 0.548, with an average of 0.238. Primers with a PIC higher than 0.5 indicated that that 12 pairs of primers had high polymorphism and could be used for diversity analysis and variety identification.Genetic parameters based on 12 pairs of SSR primers are shown in C. oleifera germplasm resources were divided into three groups  of the core collections were less than 20%, and the range coincidence rates (CRs) were greater than 80%, indicating that all core collections meet the conditions . For cort-test between the PowerCore-25 and the original collection show no significant differences in 12 fruit traits  [Prunus armeniaca L.) [C. melo L.) [C. oleifera germplasm resources was also extremely abundant. He et al. [C. oleifera in 18 areas in China, and found that the coefficient of variation of FHD and FVD was less than 20% [C. meiocarpa and one C. oleifera plants: the coefficients of variation of five phenotypic traits ranged from 23.86% to 56.94% in this study [C. oleifera and C. meiocarpa. C. meiocarpa has the characteristics of thin skin and small fruit, and its fruit phenotype was significantly different from that of C. oleifera [Genetic diversity is mainly studied at the morphological, cytological, biochemical, and molecular levels to understand the genetic diversity of species , thus pr. J. Li) , apricotiaca L.)  and melomelo L.) , substane et al.  studied than 20% . Howeveris study . The geroleifera . In thisC. oleifera plants from Guizhou and found that only FSR and PT were negatively correlated, while DSR was negatively correlated with FHD. The result was probably due to the fact that the 40 C. oleifera plants came from a unique climate in low, hot valleys, exhibiting specific genotypic performance. Furthermore, Lu Yang studied 45 superior C. weiningensis YK Li., and found no significant correlation among PT and other fruit traits and economic traits. These results may be because the studied materials belong to the thin-skinned type, and the variation among the characteristics was small [C. oleifera has great potential for genetic improvement and is important for crossbreeding.Association analysis can reveal the relationship between different traits . In thisas small . The resThe 167 germplasm resources were divided into three groups by 12 phenotypic traits. Group I had 20 germplasm resources with large fruit and thick pericarp, and their FVD, FHD, FFW, and PT were 33.05%, 21.73%, 113.78%, and 41.49% higher than those in other groups. There were 20 germplasm resources in group II, with small and thick fruit pericarp and a high seed rate. The fresh and dry seed rates were 41.34% and 33.38% higher than those in other groups. There ware 127 germplasm resources in group III, and the oil content rate of dry kernels in this group was 27.22% higher than in other groups.C. oleifera germplasm resources showed high genetic diversity. Jia et al. [C. oleifera germplasm resources, and found the average values of Na, Ho, and He were 9.1, 0.741, and 0.746, respectively. The results were consistent with our results, indicating that C. oleifera has high genetic diversity due to geographical isolation and late self-incompatibility [C. japonica and C. oleifera with 21 SSR pairs, and found that the H, I and PIC of C. oleifera were 0.2089, 0.3324, and 0.4014, respectively. The genetic parameters were smaller than those in this study, possibly because fewer research samples was used.In this study, 167 a et al.  studied tibility . Zhao ettibility  studied C. oleifera germplasm resources were divided into three groups. The clustering of 12 fruit traits also demonstrated three groups, but the two methods differed greatly regarding the classification groups. Li et al. studied 89 C. camellia genotypes and found that the same species could cluster together well, and some accessions that were grouped in the same cluster or subcluster had similar flower colors, but these groups can also exist in different colors [Hordeum vulgare L.) genotypes and found that the genotypes clustered in three sub-clusters did not show any trait-specific relationship with each other [Based on SSR molecular marker information clustering, 167 t colors . Jan et ch other . There wch other . It may ch other . The resJuglans regia L.) [Olea europaea L.) [Perilla [Core collections can preserve the genetic diversity of the original collection to the maximum extent . In thisegia L.) , which cegia L.)  used Powpaea L.)  and Peri[Perilla , core co[Perilla . PowerCoC. oleifera. Germplasm resources of C. oleifera in Guizhou were found to have a high genetic diversity and can be used to improve breeding. However, the cluster analysis showed that the environment had a substantial influence on the phenotypic and economic traits of C. oleifera. Therefore, the environment should be taken into consideration when introducing and breeding C. oleifera. In addition, the core collection of phenotype traits, economic traits, and molecular marker information was constructed in this study, and was found to be conducive to the preservation and management of C. oleifera.In this study, 12 pairs of primers were found to show high polymorphism, which can provide a reference for subsequent studies on the genetic diversity of