Resource and environmental carrying capacity (RECC) is an important basis for achieving sustainable urban development, and analysis of the relationship between regional resources and human activities is of great significance for sustainable regional development. Taking the Yangtze River Economic Belt (YREB) as the study area, this study establishes a framework for analyzing RECC based on the resource and environmental support capacity (RES) and the pressure on the resource and environment (REP), calculates the RES and REP of 110 cities in the YREB from 2009 to 2018, and analyzes the main constraints on RECC. The results show that (1) there are inter-regional imbalances in RECC within the study area, with cities that are more economically developed or at a higher administrative level usually having more severe problems with RECC. (2) The RES and REP indices of cities in the YREB show an overall increasing trend, but the relative growth rates of the RES and REP indices of cities at different levels differ. (3) The built-up area, green space in built-up areas, total gas supply, and length of sewage pipes are hindering factors for most cities to improve their RES. This study contributes to a comprehensive understanding of the current situation and changing trends of RECC in the YREB and can provide a reference for decision-making on sustainable development of the region’s large river basin. The Yangtze River Economic Belt (YREB) is the most economically developed and competitive economic belt in China, the most populous and largest river basin economic belt in the world, and an inland economic belt with global influence. In recent years, China’s rapid socio-economic development and accelerating urbanization have increased the rate of urban consumption of natural and social resources, which in turn has led to serious urban environmental degradation and resource depletion, creating a vicious cycle. The ecological space of the city is heavily encroached upon by production space and living space . ProblemResource environmental carrying capacity (RECC) is a scientific concept that measures the relationship between human socioeconomic activities and the natural environment and is an important tool for measuring and managing sustainable human development. Relative stability of the RECC is the precondition for long-term rapid development . CarryinRECC studies are analyzed from a comprehensive perspective, mainly by constructing a systematic framework containing each carrying capacity element and analyzing and studying each carrying capacity element according to a unified characterization value . RegionaCurrently, RECC research in China is an important topic. For example, Jiang Hao et al., (2016) conducted a dynamic analysis of Chengdu’s comprehensive urban carrying capacity from 2008 to 2014 from the integrated dimensions of natural resources, socio-economics, and ecological environment, where the ecological environment is a necessary factor in determining the comprehensive urban carrying capacity of Chengdu, and the influence of natural resources on the comprehensive urban carrying capacity of Chengdu was smaller . Duan PeA number of limitations exist in the current stage of research on RECC.(1)In existing studies, most scholars often divide the level of RECC index  at equal intervals or directly use the grading standards of previous literature, which can only indicate the change in the intensity of RECC and cannot indicate the carrying state of RECC . These subjective evaluation standards not only weaken the explanatory power and persuasiveness of the evaluation results to a certain extent but also cannot determine whether the pressure of human activities on resources and environment exceeds the carrying capacity of the resources and environment, which is not conducive to an accurate understanding of the sustainable state of RECC.(2)In existing studies ,37, alth(3)At present, there are few studies of the YREB RECC at the city level. Relevant studies have focused on specific regions ,39,40. HBy combining human activities with resources and environment, this paper constructs an RECC evaluation system including support and pressure factors affecting bearing capacity, which complements the existing research. The support index represents the potential of natural resources and the social public resources created by human beings to promote development. The pressure index represents the economic and social activities of human beings served by the support system. Therefore, in this paper, the resource and environment carrying capacity index (RECC), which is a single quantitative value in most other studies , can be The YREB is an economic circle near the Yangtze River that includes two municipalities (Shanghai and Chongqing) and nine provinces  . The YREWater resource data were obtained from the cities’ water resources bulletins from 2009 to 2018 (Urban Water and Lakes Agency). Other natural resource data, infrastructure data, and economic-development-related data were obtained from the cities’ statistical yearbooks from 2009 to 2018 (Urban Statistics Agency). The environmental quality data are from the cities’ annual environmental quality bulletin . Social-security-related data and environmental pollution data were obtained from the cities’ annual National Economic and Social Development Statistical Bulletin from 2009 to 2018 (city government).Regional resource and environmental carrying capacity should consider how resources, the environment, and human activities interact with each other. RECC can be examined from two perspectives: the supporting capacity of resource and environmental systems and the pollution and damage to resources and the environment caused by socio-economic activities. Within the RECC system, the support index is divided into two subsystems: the resource and environmental systems. Human social activities can be broadly summarized into three categories: economic development, social state, and environmental pollution. After combining the available data from 110 cities, 10 RES indicators and 10 REP indicators were identified . Based oIn this paper, the extreme value method is used to standardize the data and ensure that the standardized data are between 0 and 1. The specific method is as follows.In Equation (1), Evaluation of RECC must assign weights to each index. Through collation and comparison of the literature and analysis, this paper uses the analytic hierarchy process (AHP) variation coefficient method (VCM) method to determine the weights. The AHP has strong subjectivity, so most studies will use it in combination with some objective weight calculation method ,43. In tIn Equation (2), The AHP-CV method was used to determine the combined weights of the evaluation indicators .(3)WiS=β is the preference coefficient, 0 ≤ β ≤ 1. After consulting relevant experts, the coefficient β was set to 0.5. In Equation (3), Based on the normalized values of each indicator and the corresponding weights, the RES index and REP index are obtained using the linear weighting method. The calculation method is as follows .(4)Si=∑Then, the RECC index for a city is equal to the ratio of RES to REP for that region. The results of the calculation are as follows.Theoretically, according to the RES and REP indices, the RECC size can be divided into three cases, RECC < 1, RECC = 1, and RECC > 1: when RES = REP, i.e., RECC = 1, RECC is adequate for the stressful activities exerted by human beings. When RES > REP, i.e., RECC > 1, the capacity of the resource-environment system to support the population is stronger than the pressure exerted on the system by human social activities. When the pressure exerted on the support side is within the limits, there is a certain amount of residual capacity. When RES < REP, i.e., RECC < 1, the support capacity is less than the pressure exerted by human activities, and the regional RECC is overloaded. Considering the large differences in geographical location, resource endowment, environmental capacity, level of economic development, level of social development, and human activities, the RECC status of each city can be classified to guide human activities in a more targeted manner.In this paper, after referring to the literature, RECC states are classified into five types, and the classification criteria are based mainly on functions  and y=x . The speIn addition, RECC states can be divided into four types according to the size range of RES and REP indexes .The “L-H” RECC status indicates that the city has a low RES and a high REP. In the “L-H” state, although a city can achieve great economic and social development in the short term, it is not conducive to sustainable development in the long term. The best way to develop a city in an ‘L-H’ state is to gradually move to an ‘H-H’ state.The “H-H” RECC status indicates a city with a high RES and REP. A city with the “H-H” status has not only a high level of economic and social development but also good resources and environmental factors to support it and strong development momentum and development prospects. When the REP of cities with the “H-H” status gradually decreases, they will gradually shift to the “L-H” status. Each city should adjust its development status appropriately according to its own conditions and economic development level.The “L-L” RECC status indicates that the city’s RES and REP are both low. A city with the “L-L” status not only has a low level of economic and social development but also lacks the resources and environmental factors to support economic and social development, and the city’s development prospects are constrained.The “H-L” RECC status indicates that the city has a high RES and a low REP. A city with the “H-L” status has a low level of economic and social development but has a strong resource and environmental endowment. In addition to lagging development, sacrifice of economic development to protect ecological and environmental resources is a reason for the low level of economic and social development in these cities. However, this state of development is not sustainable, and cities with the “L-L” state should make full use of their resource and environmental endowments to actively explore a green and ecologically sustainable development path.A barrier degree model was introduced to explore the main barrier factors of the YREB RECC support system to effectively diagnose the pathology of the shortcomings of the YREB resource environment and to propose targeted enhancement strategies with a model structure .(7)Mij=The RES and REP indices for the 110 cities in the YREB can be calculated using the RES and REP models. Both the RES and REP indices show significant spatial variation within the YREB , while t(1)Whether in 2009 a or 2018(2)Unlike the change in the RES index, the trend of the REP index across cities in the YREB from 2009 a to 2018(3)(4)According to the RECC evaluation model, RECC indicators can be further calculated through the RES index and REP index to obtain RECC indicators. The breakdown of RECC status and city types for the 110 cities in the YREB in 2009 and 2018 is shown in As shown in (1)(2)(3)By comparing the changes in the RECC classification and status of the YREB’s 110 cities as of 2009 and 2018, it can be found that the improvement in the RECC status of Shanghai and Hangzhou is mainly due to the decrease in their REP indices, indicating that they have effectively controlled their environmental pollution levels. The improvement in the RECC status of Chongqing is due to the combination of a significant increase in the RES index and a decrease in the REP index, indicating that its resource security capacity and environmental management level have been significantly improved and its environmental pollution situation has been improved. The deterioration in the RECC status of Wuxi and Suzhou is caused by the decrease in the RES index and the significant increase in the REP index, which indicates that they have not only under-invested in their resource security capacity and environmental governance level but also experienced deterioration in their environmental pollution situation. These cities must take necessary measures and increase their financial investment to improve this disadvantage.Due to the dynamic economic and social development of the YREB, the interaction between human activities and resource and environmental systems in the area is frequent and complex. Factors including changes in population, changes in industrial structure, intensity of urban construction, type and size of land use, paving of urban roads and other infrastructure development, improvements in resource use efficiency, and significant improvements in environmental technologies inevitably have a significant impact on the city’s RES and REP indices, so the RES and REP indices of other cities are also in a dynamic state of change. However, as these cities are at a relatively low level of development, their own resource and environmental potential can still support their economic and social development consumption; therefore, their RECCs have not yet reached an overload state. These cities must pay attention to improving their resource security capacity and environmental protection in the future development process and follow the path of sustainable development.Notably, while cities other than Shanghai, Chongqing, Chengdu, and Suzhou are all L-L cities, some cities, such as Wuhan and Wuxi, are in a critical position and are trending towards L-H cities. If these cities fail to pay attention to improving resource efficiency and environmental protection efforts while developing their economies and shift from pursuing high economic growth to a model of high-quality economic development, their economic and social development will not be sustainable. Their governments must promptly adjust their development paths, reform their economic development models, and follow a sustainable development path to achieve simultaneous growth in the RES and REP indices and eventually develop towards H-H cities.By analyzing the current barriers to RES indicators in the RES system and identifying the shortcomings of RES indicators, targeted policies and recommendations can be made for the future development of cities. Based on the barrier degree calculation method, it is possible to derive the barrier degree of each indicator supporting large cities in 2018. (1)(2)(3)Obstacle factors, ratio of good air quality (S7) g, ratio In this paper, the RECC system is divided into two parts: the RES index, which reflects the resource security capacity and the environmental condition and governance level; and the REP index, which reflects the pressure of economic activities, social security, and environmental pollution on the resource and environmental system. The indicators are weighted: the entropy weighting method is used to determine the weight coefficients of each indicator and the linear weighting method is used to calculate the RES index and REP index. In this paper, the RECC index is the ratio of RES index and REP index rather than the simple linear weighted sum . In addition, combined with the coupling curve, it can not only calculate the size of the RECC but also judge the status of the RECC (overload/surplus) so as to more accurately formulate targeted development strategies for cities with different RECC statuses.This paper finds that, among the 110 cities in the YREB between 2009 and 2018, the higher the administrative level of a city is, the higher the RES index and REP index of that city. Considering that, in China, the higher the administrative level of a city is, the better its level of economic and social development in general, it can, therefore, be concluded that, the more developed a city is, the more supportive it is of its resource and environmental system and the more pressure human activities place on it. This is similar to the findings of the study by Zhang Fei et al. on RECC in large cities in China. In that study, they found that Chinese megacities face greater pressures on the resource environment. The study by Shang Yongmin et al. on the RECC of cities in the YREB also shows that the economically developed eastern coastal areas face higher resource and environmental pressures than the western inland areas. Other studies have also found that the more developed cities, such as Shanghai and Guiyang, perform better between coordination of economy and RECC ,50. SomeThis paper differs from other existing studies in that it finds that the gap in the RES index between the municipalities directly under the central government, Shanghai and Chongqing, is becoming wider, while the gap in the RES index between different provincial capitals or different prefecture-level cities is decreasing. In contrast to the change in the RES index, the gap in the REP index between the municipalities directly under the central government, Shanghai and Chongqing, is decreasing, while the gap in the RES index between provincial capitals and within prefecture-level cities is increasing. The change in the RECC index is more complex as it is calculated by combining the RES index and the REP index. According to the research in this paper, there is a large difference between the RECC of a municipality or provincial capital city in the YREB and its neighboring prefecture-level cities. These cities are the central cities of the surrounding region, with higher levels of economic and social development and greater pressure on the resources and environment caused by human activities, so they are mostly in the resource and environmental overload zone. Moreover, cities in the eastern part of the YREB are under greater pressure on RECC than cities in the west. This phenomenon reflects the reality of China’s economic and social development, where urban development is being severely challenged by spatial imbalance, with each province often having only one development center, the provincial capital city, which can absorb the province’s resources for priority development. Furthermore, due to their location, cities in the east usually have a better level of economic and social development than cities in the west, and the combination of these factors creates uneven levels of urban development between and within provinces. Therefore, the government has realized intra-provincial economic linkages to accelerate development of prefecture-level cities, thereby easing the pressure on resources and the environment in the central cities. In addition, the RECC statuses of Shanghai, Chongqing, and Hangzhou all improved, while Suzhou and Wuxi experienced a deterioration in their RECC status.Using a barrier degree model, this paper finds that built-up area, built-up green space, total gas supply, and length of sewerage pipes are the main barriers faced by most cities in the YREB, except Shanghai. Cities must improve urban infrastructure, optimize social resources, and increase investment in environmental management. Shanghai, on the other hand, due to its smaller area and higher level of development, is constrained mainly by the total water resources, sown area, built-up area, and road mileage and needs to focus on improving the green efficiency of resources .Combined with the above analysis, the following policy suggestions are formulated to provide a relevant basis for the sustainable development of the YREB.For municipal authorities and provincial capitals, their economic development level has reached a high level, so the relationship between human activities and resources and environmental systems in these cities is extremely tense. These locations should implement strict water resource management and rational land use planning in the future, and large-scale resource redeployment projects across regions, such as south–north water transfer and west–east gas transmission, still need to be built. In addition, these cities should increase the supply of social resources, such as urban green space and urban road areas. To solve the problem of resource scarcity, cities must change their production methods and lifestyles to improve the efficiency of resource use. Advanced cities should improve green innovation capabilities , developFor the prefecture-level cities, their economic development is not good enough, which strongly necessitates economic development. Therefore, they have to make development plans in advance, prioritize development of resource-saving and environment-friendly industries, and focus on development of the circular economy. The government should strengthen investment in environmental management and improve coverage of technologies such as water purification technology, industrial sulfur dioxide treatment technology, industrial wastewater treatment technology, and domestic waste treatment technology to effectively reduce sewage discharge and waste generation. The city should pay attention to reducing the impact on and damage to existing resources and the environment while developing economically and actively exploring the path of sustainable development.This paper evaluates only some of the cities’ resources, environment, and economic and social activities and does not address trade and other mobility factors between cities and regions. In addition, arable land area, forest area, food production, energy consumption, and some other factors were not considered due to problems in data collection. The driving forces behind changes in RECC in cities have not been studied and need to be assessed in future studies.(1)Within the YREB, there are regional imbalances in RECC such that, the more economically and socially developed a city is, or the higher its administrative level, the more serious its RECC problems are. This is reflected mainly in the fact that municipalities and provincial capitals are generally the most heavily loaded cities in the region in terms of RECC, and most of them are already in a state of overload, with the pressure of their human and social activities exceeding the capacity of the local resources and environmental services.(2)Both the RES and REP indices of cities in the YREB show an overall increasing trend, but the relative growth rates of the RES and REP indices vary depending on the administrative level of the city. Among them, the difference between RES indices of different municipalities directly under the central government increases and the difference between REP indices decreases; the difference between RES indices of different provincial capitals or different prefecture-level cities decreases and the difference between REP indices increases.(3)The area of built-up areas, the area of green areas in built-up areas, the total amount of gas supply, and the length of sewerage pipes are the main limiting factors for cities other than Shanghai to improve their RES; the total amount of water resources, the area of sown seeds, the area of built-up areas, and road mileage are the main limiting factors for Shanghai to improve its RES.Based on the RES and REP coupling mechanism, this paper examines the RECC situation and trends of 110 cities in the YREB from 2009 to 2018. In addition, the main obstacles to the cities of the YREB in 2018 to improving their support capacity are analyzed. The main conclusions are as follows:


Information on genetic variation within the chloroplast genome is of great interest to ensure an efficient conservation strategy that takes into account the preservation of the various maternal lineages of Tunisian barley.In Tunisia, barley local landraces are still cropped for human and animal consumption in some subsistence farming systems under marginal and stressed conditions. These high-value genetic resources present a potential source of resistance genes to biotic and abiotic stresses useful for both national and international breeders. Actually, they are represented by threatened small populations, which face a high risk of genetic erosion and progressive substitution by modern varieties. In this study, the genetic diversity of 60 Tunisian barley landraces was assessed using six chloroplast microsatellites. All loci were found polymorphic, with 2 or 3 alleles per locus. Thirteen alleles were detected across the studied sample, which were combined into 8 haplotypes, giving a haplotype diversity (Hd) of 0.847. High punctual and haplotype genetic diversity was observed for Tunisian barley landraces when compared to other germplasms from other regions of the world. The genetic structure analysis revealed two major clusters of Tunisian barley landraces, which confirms their multiorigin. This result was corroborated by the median-joining network showing the genetic relationships among the eight detected haplotypes. The AMOVA analysis revealed that 83% of the genetic variation is between populations, which requires the  Hordeum vulgare L., Poaceae) is the fourth main cereal crop, behind only rice, wheat, and corn [Hordeum spontaneum K. Koch, in the Fertile Crescent about 10,000 years ago [Hordeum vulgare was considered a model species for plant genetic studies for both basic and applied research due to its diploid nature (2n = 14), its reduced number of chromosomes, and its ease of crossing [Barley  molecular markers. The genotyped genetic resources are representative of the main distribution areas of this crop in Tunisia.The seeds of 60 accessions of Tunisian local barley representing 11 landraces populations were collected from different regions belonging to various bioclimatic stages to serve as plant material for this study. The bioclimatic characteristics of the investigated regions are described in μl as follows: 1 μl PCR buffer, 0.2 μl DNTPs (25 mM), 0.7 μl MgCl2 (25 mM), 1 μl DNA (10 μM), 1 μl primer F and 1 μl primer R (10 μM), 0.1 μl Taq polymerase (0.5 U), and 5 μl of distilled water. The amplified DNA fragments were separated by electrophoresis using a 2.5% agarose gel. A 100-bp DNA ladder was used to determine the allele's size.Molecular analysis of the chloroplast genome of 60 Tunisian barley accessions was carried out using 6 cpSSR markers , previouVarious allelic and haplotype genetic parameters were estimated across the 60 barley landraces. The software GenAlEx 6.5  was usedThe genetic relationships and genetic structure analyses among the studied barley populations were assessed. The UPGMA analysis was carried out using Darwin software v.6.0.021, applying dissimilarity indexes calculated from single data . MoleculIn this study, 6 cpSSR loci were used to assess the chloroplast genetic diversity of 60 local Tunisian barley accessions representing the distribution area of this species in Tunisia. The obtained results showed that the loci are all polymorphic. The genotyping of the total sample allowed the detection of 13 alleles with a mean value of 2.167 ± 0.167 alleles per locus. The effective number of alleles per locus varies from 1.220 for the locus hvcptrnS1 to 2.410 for hvcptrnS2, with a mean value of 1.814 ± 0.165. Shannon's information index varies from 0.325 to 0.960 with a mean value of 0.637 ± 0.084. The genetic diversity indices h and uh vary from 0.180 and 0.183 (hvcptrnS1) to 0.585 and 0.595 (hvcptrnS2), respectively .Due to the haploid nature of chloroplast DNA, a unique assembly between the six genotyped microsatellite loci was considered a distinct haplotype. The thirteen identified alleles at the 6 loci were combined into 8 haplotypes with haplotypic frequencies that range between 0.050 for haplotype 8 and 0.280 for haplotype 7 . A haploIt is noted that one unique haplotype was detected for each of the five barley populations Beja (haplo-3), Bizerte (haplo-1), Gabes (haplo-7), Kasserine (haplo-8), Kef (haplo-6), and Zaghouan (haplo-2), while two haplotypes were observed for each of the remains populations . One private haplotype was observed for each of the populations Beja (haplo-3), Bizerte (haplo-1), Kef (haplo-6), Medenine (haplo-4), and Siliana (haplo-5). The effective number of haplotypes (Ne) ranged between 1  and 2 haplotypes (Sousse). The haplotype genetic diversity (Hd) varied among the 11 populations between 0 and 1, with a mean value of 0.260 .In order to clarify the genetic relationships among the 8 detected haplotypes (H) in Tunisian barley landraces, a median-joining network was constructed . This anAn UPGMA dendrogram was constructed to evaluate the genetic relationships between the 11 studied populations of Tunisian barley landraces . The obtP value = 0.001) among the local barley populations.The analysis of molecular variance (AMOVA) was achieved to analyze the amounts of genetic diversity between and within populations . Based oThe molecular characterisation of Tunisian barley landraces showed that the six used loci are all polymorphic across the genotyped barley sample and allowed the detection of 13 total alleles. Two to three alleles per locus were detected, with a mean value of 2.16 alleles per locus. The detected polymorphism in this study is considered as important given the haploid nature of the studied loci and the mating system of barley. The obtained results corroborate previous studies, which focused on chloroplast SSRs diversity in the barley genome and showed that two to three alleles per locus were detected , 21, 28.A total of 8 haplotypes were identified in Tunisian barley landraces based on the six cpSSR loci. The analysis of the haplotype diversity among the studied populations showed that the number of haplotypes per population varies between one and two haplotypes. A high haplotypic genetic diversity (0.847) across the studied 60 landraces is detected. This level is higher than values previously reported for other studies interested in barley landraces. A haplotype genetic diversity of 0.330 was recorded for Turkish barley landraces , while PThe detected variation in the amounts of haplotype diversity among the studied samples of barley landraces may be explained by various factors, namely the number of analysed landraces, the geographic and bioclimatic characteristics of the prospected areas, and the domestication status of the analysed plant material. Indeed, the domestication process of barley from its wild progenitors resulted in a dramatic reduction in its genetic diversity. Lower genetic diversity amounts for barley landraces compared to wild accessions and for cultivars compared to landraces were previously highlighted by applying chloroplast molecular markers . Thus, tThe genetic structure analysis of the studied germplasms revealed two main clusters. Two main clusters were also obtained when five barley cpSSRs were applied to investigate the genetic relationships among 186 barley accessions representing cultivated and wild germplasms that originated from the main distribution areas of this crop except for the Far East region . The obsThe phylogenetic relationships among the eight detected haplotypes evaluated by a median-joining network revealed a complex genetic relationship pattern. This analysis highlighted three frequent haplotypes and other minor haplotypes that may have evolved through mutation events in these major haplotypes over time. Indeed, the most frequent and shared haplotypes are generally considered to be the most ancient ones, and minor haplotypes for chloroplast SSRs appeared generally following the stepwise mutation process .The obtained results showed considerable allelic and haplotype genetic diversity for the chloroplast genome of Tunisian barley landraces. The detected molecular polymorphism in these threatened local landraces may be lost over time if any conservation measures are not achieved. The conservation of the genetic resources of barley, especially ancient landraces, is required to maintain the maximum allelic richness that can be exploited in the future to enhance the quality and yields of this species . Indeed,The AMOVA analysis of the studied landraces revealed that 83% of the observed genetic variability is among populations. For agricultural crops, human selection and management in addition to natural selection are considered the main factors that define the patterns of intraspecific genetic diversity and differentiation . The obsex situ conservation in gene banks of the seeds of all the detected maternal lineages is of great interest for a further improvement strategy for barley. Furthermore, in situ conservation measures are required as a complementary conservation strategy to allow the evolution of these valuable landraces in the various edaphic and bioclimatic conditions of Tunisia to maintain their genetic polymorphism. It is necessary to pay specific attention to the conservation of landraces with private and rare alleles and haplotypes.Chloroplast genomes were reported to contain genes relevant to the expression of various agronomically important traits such as cytoplasmic male sterility, plant growth, and response to adverse conditions –34. Thusex situ conservation of local germplasms of cereal crops, including barley landraces, has been started over the last decade in Tunisia, more urgent efforts are needed. In addition to the static ex situ conservation tool, the establishment of dynamic in situ collections of barley landraces with various edaphic and bioclimatic characteristics is required to ensure the preservation and evolution of their adaptation potentials in the face of changing environmental conditions.While the challenge of the The present study illustrates the efficiency of a set of informative chloroplast sequence repeats (cpSSRs) to assess the variation of genetic diversity, structure, and differentiation among various gene pools of barley landraces. A high haplotype genetic diversity was detected for Tunisian barley landraces, which are clustered into two main groups. These cpSSRs present complementary molecular tools to nuclear molecular markers to evaluate the maternal lineage diversity and classification of barley landraces. The characterisation of the cytoplasmic genome of Tunisian local barley landraces is of great interest for the establishment of conservation strategies for this cereal crop, which will be useful as a source of specific traits in future breeding programs.


Steady progress in the field of psychiatric genetics, generating new and fascinating insight into the genetic and phenotypic complexity of schizophrenia and other serious mental illnesses, have created an increased need of psychiatric genetic counseling. It is a crucial aspect of psychiatric clinical practice to ensure a balanced approach that takes into consideration genetic and environmental risk factors and ongoing education in this rapidly developing field is essential. Genetic counseling will be increasingly important to assist patients with schizophrenia and their families to make a meaningful informed decision about genetic testing. It will preempt unrealistic expectations, discrimination and stigma related to incomplete understanding of genetic test results in the psychiatric context. Major psychiatric disorders, including schizophrenia are complex, polygenic disorders. Genetic and environmental factors contribute to the susceptibility of an individual to develop schizophrenia . It is wThe identification of genes that contribute to the risk of developing a mental disorder and influence the course of the illness is the goal of molecular genetic studies in the field. Vulnerability to a mental illness is a genetically complex trait that emerges from the interaction of multiple genes. This combination of genes do not fully control the risk of developing a mental illness and is dependent on the interaction with non-genetic factors . There hAdvances in psychiatric genetics has unraveled the genetic architecture of mental illness. The challenge psychiatrists face is the translation of the genetic findings into clinical practice to improve the lives of patients and families. Psychiatric genetic counseling (PGC) is a minor sub-specialty of genetic counseling that is not routinely provided for psychiatric conditions and there is a lack of consensus about the best way to deliver it to patients . It is ePsychiatrists regularly have to answer questions about heritability, genetic risks and the role of genetic testing in diagnosis or treatment selection . To bettSpecific details about the exact gene, receptor or neurotransmitter involved are not required during the process of PGC, but the information should be detailed enough for a patient or family to make an informed decision about genetic testing. Some of the important findings to take into consideration will be summarized here, without going into specific gene, receptor or molecular details. There has been a lot of progress in psychiatric genetics due to genome-wide association studies (GWAS) with the focus on identifying the underlying biological mechanisms contributing to disease. It includes investigation of polygenic mechanisms and examination of copy number variants (CNVs) and potential clinical applications for polygenic risk scores that calculate the joint contribution of common variants. The aim is to identify the genetic influences on mental disorders and rare single-nucleotide variants (SNVs) have been associated with schizophrenia and other psychiatric disorders .de novo and that CNVs predispose to a variety of psychiatric disorders with limited diagnostic specificity. CNVs tend to be expressed in the developing brain suggesting a common neurodevelopmental origin for many psychiatric disorders indicating that primary prevention would need to start in childhood  variants and the evidence points to important roles for both. The tissue and cell types in which the newly identified risk variants are most biologically active has been found to be in brain tissue. On cellular level, neuronal cell subtypes have been most closely associated with the risk to develop schizophrenia . IndividEpigenetics is defined at the study of chemical changes at the cellular level that alter gene expression without altering the genetic code and is a significant consideration for PGC , 16, 17.Like many mental disorders, schizophrenia have a robust genetic component reflected in the high degree of heritability. In the simplest terms, it can be explained to a family that if a family member has been diagnosed with major psychiatric disorder it increases other family members' risk of developing a similar disorder. There is an overlap in common genetic vulnerability markers between schizophrenia and other disorders, mostly bipolar disorder, that should be taken into consideration. Individuals can also have a higher risk to develop a mental disorder due to specific genetic mutations or polymorphisms without a family history , 9.In patients with schizophrenia there will be one or more than one gene that interact with each other and interact with a variety of environmental factors, following a non-mendelian pattern of inheritance. The genetic heterogeneity means that the phenotype is not certain to be an indication of the genotype and several genetic mechanisms like mutation at separate loci or different mutations at a locus can lead to the same or similar phenotype . MultifaThe probability estimates of developing an illness may be useful for an individual or family members, but recurrence risk estimates for schizophrenia can be difficult and complex. The recurrence risk estimates are based on empiric risks and on statistical probabilities of particular study populations. The genetic risk will not translate to a specific family where the interaction with environmental factors may increase or lower the risk. The difficulty to predict the probability of schizophrenia developing in a specific family member, because of the complexity of multiple genes interacting with environmental risk factors should be addressed during PGC , 8. To dThe genetic architecture of these disorders is becoming clearer and schizophrenia has been found to be 81% heritable with single nucleotide polymorphism (SNP) based estimates currently explaining 24% of heritability .The polygenicity and genetic heterogeneity of schizophrenia, and how variants that influence illness may be widely distributed in the general population should be made clear during PGC. Several environmental factors act throughout life to influence the likelihood of the development of schizophrenia and the influence of genes and environmental factors combined will have to exceed a threshold for an individual to present with schizophrenia. The environmental influences are not causal factors and it can be grouped into early development, proximal factors and onset factors. Early development factors can include obstetric complications and advanced paternal age, while proximal factors refer to social adversity, migration or urbanicity experienced by the individual. The onset can be triggered by substance use, trauma or social adversity . DespiteGenetic testing for schizophrenia has not been widely implemented and psychiatrists do not routinely use genetic testing as part of diagnostic practice and more training is needed in this field . The maiUnderstanding the etiology and familial recurrence risk: Genetic counseling in psychiatry should endeavor to help people understand the origin and role of genetics in psychiatric disorders . The limIdentifying protective factors and improving health-related behaviors: The discussions should also address how certain environmental factors, for example extreme stress, infections, malnutrition or substance use, amongst others, may trigger the onset, relapse or worsening of symptoms . PGC shoProviding information to facilitate decision-making and to address stigma: The counselor should address potential harmful effects of genetic testing. These might include family conflicts, uncertainty about the probability of developing an illness and stigmatization. It can potentially influence important life decisions about a life partner and reproductive choices . OverrelPsychotherapeutic aspects: The process of PGC has been described extensively and consist broadly of information gathering  and provision of information and support . The potThere are many diverse and rapidly changing ethical concerns in psychiatric genetics. These concerns, especially in the field of research, big databases and direct-to-consumer-testing are vast. The brief discussion here will focus on some ethical aspects that should be considered during PGC in the clinical setting . The decDuring the process of PGC issues of beneficence and non-maleficence should be considered. The potential harm should be balanced with the expected benefit from doing a specific genetic test and individuals should be supported to make a meaningful autonomous choice. These choices should be based on the individual values of the patient in the context of the family history, without any undue influence . The comWhenever genetic tests are requested the implications for family members and the risk of incidental findings are important considerations. It should be discussed beforehand if an individual would want to learn of any incidental findings. As far as possible, families should participate in any decision that might affect them. Due to fast developments in the field of genomic medicine, continuing education should be combined with a dynamic process to provide information to enhance informed consent .Misuse of genetic data to discriminate against individuals applying for insurance or in employment remain a concern . ConcernWith the rapid and major developments in the field of genetics and genomics, it is important for psychiatrists to ensure that they remain up to date with the latest developments. PGC cannot be left only to specialized genetic counselors. It is a crucial aspect of psychiatric clinical practice to ensure a balanced approach that takes into consideration genetic and environmental risk factors. PGC will be increasingly important to assist patients to make a meaningful informed decision about genetic testing and will preempt unrealistic expectations, stigma and discrimination related to incomplete understanding of genetic test results in the psychiatric context.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.CK and GZ were involved with the original presentation that this perspective article is based on. All authors contributed to the write up of this perspective article.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Elsholtzia stauntonii Benth. (Lamiaceae) is an economically important ornamental, medicinal and aromatic plant species. To meet the increasing market demand for E. stauntonii, it is necessary to assess genetic diversity within the species to accelerate the process of genetic improvement. Analysis of the transferability of simple sequence repeat (SSR) markers from related species or genera is a fast and economical method to evaluate diversity, and can ensure the availability of molecular markers in crops with limited genomic resources. In this study, the cross-genera transferability of 497 SSR markers selected from other members of the Lamiaceae  to E. stauntonii was 9.05% (45 primers). Among the 45 transferable markers, 10 markers revealed relatively high polymorphism in E. stauntonii. The genetic variation among 825 individuals from 18 natural populations of E. stauntonii in Hebei Province of China was analyzed using the 10 polymorphic SSR markers. On the basis of the SSR data, the average number of alleles (NA), expected heterozygosity (HE), and Shannon’s information index (I) of the 10 primers pairs were 7.000, 0.478, and 0.688, respectively. Lower gene flow (Nm = 1.252) and high genetic differentiation (Fst = 0.181) were detected in the populations. Analysis of molecular variance (AMOVA) revealed that most of the variation (81.47%) was within the populations. Integrating the results of STRUCTURE, UPGMA (Unweighted Pair Group Method with Arithmetic Mean) clustering, and principal coordinate analysis, the 825 samples were grouped into two clusters associated with geographical provenance (southwestern and northeastern regions), which was consistent with the results of a Mantel test . Overall, SSR markers developed in related genera were effective to study the genetic structure and genetic diversity in geographical populations of E. stauntonii. The results provide a theoretical basis for conservation of genetic resources, genetic improvement, and construction of a core collection for E. stauntonii. Lavandula angustifolia) (Hedeoma patens)  , while m patens) . ElsholtProvince . Elsholtproducts . Consequcticides . With itar plant . Its braar plant . Furtherar plant . Previoul traits , cultival traits , and extl traits . Little l traits  and to il traits . TherefoScutellaria baicalensis . The DNA quality was evaluated by 1.0% agarose gel electrophoresis. DNA quantification was conducted with a NanoDrop 2000 spectrophotometer  and the DNA concentration was adjusted to 20 ng/μL.E. stauntonii samples from different populations  × 100. Similarly, the percentage transferability of SSR markers classified by repeat motifs (di-/tri-/tetra-/penta-/hexa-/complex-nucleotide) was estimated. Population diversity and population structure of NA), number of effective alleles (NE), Shannon’s information index (I), observed heterozygosity (HO), expected heterozygosity (HE), inbreeding coefficient (Fis), gene flow (Nm), and genetic differentiation index (Fst). The polymorphic information content (PIC) value and Hardy–Weinberg equilibrium (HWE) for the loci were calculated using PowerMarker version 3.25 , 15 independent runs were performed with a burn-in period of 200,000 iterations followed by 1,000,000 Markov chain Monte Carlo repetitions. The ΔK method  was determined; an individual with a Q value higher than 0.80 was considered to have a single genetic component . Nei’s genetic distance (DA)  was conducted using Arlequin version 3.5  to estimon 2.3.4 . For eacK method  was implK method  program nce (DA)  among thnce (DA) . To explnce (DA) . The dennce (DA) . Based once (DA) . A clust1. The Mantel test was performed to analyze the correlation between geographic and genetic distance of different populations using the “ggpubr2” and “ggplot2”  successfully amplified genomic DNA of ted size . Transfelaria L. . The tralaria L. . The SSRE. stauntonii  or showed very low levels of polymorphism (two primers) based on capillary electrophoresis results . Ten  of eight markers was lower than the expected heterozygosity (HE), and the average inbreeding coefficient (Fis) was positive. The PIC values ranged from 0.191 to 0.868, with a mean of 0.429. Eight SSR markers showed moderate or high polymorphism levels (PIC > 0.25) and two markers (ssps344 and KNUPF67) showed low polymorphism levels (PIC < 0.25) among the tested E. stauntonii populations. All SSR genotyping data for the 10 loci showed strongly significant deviation from the HWE (p < 0.01)  .E. stauntonii was observed  to 3.900 (XM), and from 1.406 (ZQ) to 2.532 (CX), with means of 3.267 and 1.971, respectively. The average HO and HE were 0.359 and 0.388, ranging from 0.253 (XW) to 0.478 (CX), and from 0.260 (ZQ) to 0.506 (CX), respectively. The I value ranged from 0.450 (ZQ) to 0.887 (CX), with an average of 0.688. The highest level of genetic diversity was detected in CX, whereas XW and ZQ showed the lowest genetic diversity  to 0.199 (between populations CX and ZQ)  (The pairwise  and ZQ) . The pai and ZQ) . In gene< 0.001) .E. stauntonii populations, the co-ancestral relationship among populations was analyzed based on a Bayesian assignment model. The Q value of 680 individuals, accounting for 82.42% of the total samples, was greater than 0.8, indicating that most individuals had a relatively simple genetic background  formed a third group , which might reflect that the samples were collected from natural populations affected by complex environmental factors and human activities   (Perilla frutescens (0.582)  , which ws (0.72)  and Peri (0.582) . The gen (0.582) . For E. llection . Furtherllection .NA (7.000) and NE (1.971) observed in the present study were slightly higher than the values  for 12 Plectranthus edulis populations evaluated using 20 SSR markers   (E. stauntonii (HO = 0.360) has higher genetic diversity than Salvia fruticosa (HO = 0.307) (Hedeoma piperita (HO = 0.3)  (E. stauntonii samples showed moderate genetic diversity. The observed heterozygosity in the present study was lower than the expected heterozygosity (HE = 0.478), indicating that the number of heterozygotes was lower than the predicted value (Fis = 0.192) in the population was positive and high, which similarly suggested there was an overabundance of homozygotes   and Jugl = 7.51) . Compare= 0.307)  and HedeO = 0.3) , but a h = 0.63) . These red value . The aveozygotes . The rep species .E. stauntonii (Nm = 1.252) was moderately low, lower than that reported for Eragrostis tef (Nm = 4.742) (Mucuna pruriens (Nm = 7.48) (Fst = 0.181) (E. stauntonii populations (18.53%) was higher than that of many other plant species, such as Origanum vulgare subsp. hirtum (8%) (Allium sativum L. (12.29%)  showed that increase in geographic distance reduced gene flow. Therefore, geographic distance may be the principal factor for genetic differentiation in E. stauntonii.Genetic differentiation is an important index of the genetic structure of populations within a species . In the = 4.742)  and Mucu = 7.48) , but it = 0.181) . The low= 0.181) . Notablytum (8%)  and Alli(12.29%) , suggestE. stauntonii. Examination of the topography and geomorphology in Hebei Province indicated that the genetic structure of E. stauntonii was delimited according to geographical location and topographic barriers: one genealogical lineage each was located near the Taihang Mountains and the Yanshan Mountains. Previous studies have shown that geographical barriers play a crucial role in preventing plant dispersal , which also indicated the role of geographical barriers in genetic differentiation. The Taihang and Yanshan mountains may be responsible for maintaining the major southwest–northeast split of E. stauntonii populations by hindering animal-mediated gene exchange. Thus, natural adaptation and geographical barriers may explain the differentiation of the two regions. Moreover, in the present analyses, each group was more or less intermingled with lineages of the other group, indicating that there was a certain level of gene flow between the populations, which has also been reported in Tectona grandis (In the UPGMA clustering analysis, a certain deviation was observed in the clustering of individuals and of populations, which might be due to the high degree of gene exchange between populations. Overall, the clustering results observed with STRUCTURE, PCoA, and UPGMA were essentially similar, indicating that there might be two gene pools of ispersal , for exaispersal . In this grandis . The lev grandis . These fE. stauntonii. Analysis based on 10 transferable SSR markers showed that, although the genetic diversity in 18 geographical populations of E. stauntonii was slightly lower than that of certain other species in the Lamiaceae, the genetic diversity was generally rich and the populations showed a high degree of genetic differentiation. STRUCTURE, PCoA, and UPGMA clustering analyses of all individuals resolved two genetic clusters, which might reflect geographical barriers to gene flow. By screening the study materials with superior traits and rich genetic variation, some individuals in the present experiment are expected to be utilized in the future improvement programmes of E. stauntonii to meet the increasing demand from the floriculture, essential oil, pharmaceutical, and other industries. The information obtained from the analysis of genetic diversity and population structure provides a foundation for the conservation, management, and utilization of E. stauntonii.In this study, SSR markers from seven genera of Lamiaceae were transferable to The original contributions presented in the study are included in the article/YL and BC conceived and designed the experiments and obtained the funding. CZ, XL, and HZ conducted the experiments. CJ and BC contributed to the investigation. CZ, LH, and ML collected and analyzed the data. CZ wrote the manuscript. YL provided suggestions and comments for the manuscript. All authors read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Genetic counsellors are an internationally recognised group of highly skilled healthcare professionals with training and expertise in genomic medicine and counselling skills , 2. The The delivery of genomic testing had, until relatively recently, been the domain of clinical genetics services; however, such testing has now become ‘mainstream’, in that clinicians working across any discipline of healthcare can order certain genomic tests relevant to their patient. Such mainstream delivery of genomic medicine provides infinite opportunities for the profession of genetic counselling to support, mentor, train and offer clinical supervision to non-genetics colleagues who are utilising genomic technologies.Genetic counsellors have expertise and skill that can be applied in every stage of the patient pathway within genomic medicine . There iGenetic counsellors are skilled and trained in: calculating genetic risk, explaining inheritance patterns, ordering genomic testing, interpreting variants, arranging medical and/or diagnostic testing as well as testing of relatives, predicting risks of genetic disease, referring patients for appropriate disease screening and handling all the consequent psychosocial and ethical issues raised for individuals and their families. They can also act as on-call specialists for urgent referrals to the clinical genetics service and triage referrals into the service. They are the main health professional group who ‘takes care of the family’, facilitating family communication, coping and adjustment as well as cascading information and arranging testing of at-risk relatives. A key role that distinguishes genetic counsellors is that of being patient advocates both within a clinical genetics setting but more so in their work across specialities where the best interest of the patient and/or the family may find itself in conflict with what is deemed ‘routine practice’.In the UK, a clinical consultation usually takes, on average, 45 mins (with a range of 30–90 min based on the needs of the patient). Within a full day clinic, the number of patients seen will vary depending on the clinic type, patient concerns and complexity of the cases. However, on average, genetic counsellors may see between 7 and 10 patients per full day clinic. Genetic counsellors are skilled in delivering their consultations face-to-face, on the telephone or increasingly by virtual (video) clinics.Classically, genetic counsellors work in tandem with their clinical geneticist and clinical laboratory scientist colleagues as part of a tertiary clinical genetics service (many of which have been renamed Genomic Medicine Services) in the UK. Continuation of this model remains vitally important; however, an uncoupling of these roles has already started for some genetic counsellors, who increasingly also work in other settings. ‘Uncoupling’ should not be read as ‘no longer working together’. It is important for genetic counsellors working outside of clinical genetics to be able to retain a link to this service for multidisciplinary team input, line management from consultant genetic counsellors and counselling supervision.Within ‘mainstream medicine’, i.e. outside specialised clinical genetic services, genetic counsellors can support and advise non-genetics clinicians to deliver genomic medicine. This includes supporting clinicians to order the most appropriate test for their patient and providing education so that colleagues understand the nuances of informed consent relevant to genomic testing. This ensures that resources are targeted appropriately, minimising the cost, delay and negative impact of inappropriate genomic testing. Genetic counsellors currently play a role in helping to discharge inappropriate recall of patients from disease screening programmes, by refining genetic risks and re-targeting screening to those most at risk of developing disease . They also sit as the genetics expert in multidisciplinary team meetings across hospital settings, offering advice on referral pathways, interpretation of genomics results, and patient management. Across the UK there are already genetic counsellors practicing autonomously within non-genetics specialisms, e.g. within the ophthalmology, cancer, endocrine, psychiatry, fertility, ENT and dermatology clinics.There are also ‘mainstream’ genetic counsellors who, whilst attached to their local clinical genetics service, work directly with their non-specialist colleagues across a hospital who are delivering genomic medicine services that do not need the specialist input of clinical genetics. Genetic counsellors have been welcomed by many disciplines outside of clinical genetics, and often act as the link between services, ensuring and supporting the competent delivery of genomic medicine by non-genetics healthcare professionals. Models of genetic counselling delivery are evolving, and this is a reflection of the adaptability and transferability of the genetic counselling skill set.In many cases, results of genomic tests done within mainstream settings may have implications for disciplines outside of the medical specialty ordering the test. For example, a variant predisposing a patient to inherited heart disease may be identified as an ‘additional looked for finding’ in a patient in a cancer clinic; or an adult patient in a lipid clinic may have a variant identified that predicts a risk of disease to offspring, and so reproductive options need to be explored. In these situations, genetic counsellors working in mainstream settings can facilitate onward referrals or co-ordinate/manage the cases themselves, for example for cascade genetic testing of relatives, prenatal diagnosis or preimplantation genetic diagnosis (PGD).In the majority of cases, the results and information obtained from genomic analysis have implications not only for the index case but their relatives. For every new genetic diagnosis made in a non-genetics healthcare setting, the clinician who orders the test and delivers the results, should have a conversation about the impact of these for relatives. Given that non-genetics clinicians would not routinely care for at-risk, healthy relatives, the future management of such ‘patients in waiting’ can be managed via a genetic counsellor.In addition to established clinical practice, the genetic counsellor role has also evolved and now genetic counsellors practice as leaders in research, policy, education , private practice, service management and professional regulation.The genetic counselling profession established a professional accreditation process overseen by the Genetic Counselling Registration Board (GCRB), with recognised routes of entry into the profession and applicants required to deliver a Masters-level portfolio of evidence to demonstrate professional competence. In 2019 the GCRB transitioned under the auspices of the Academy for Healthcare Science (AHCS) and is now an integral part of the Academy’s regulatory function. The AHCS maintains the UK register of Healthcare Science practitioners which is accredited by the Professional Standards Authority for Health and Social Care (PSA) on an annual basis. Genetic counsellors are currently assessed by the GCRB for inclusion on the AHCS accredited register and are expected to meet the professional standards of competency and conduct as set by the GCRB and approved by the AHCS. Any concerns or complaints made about a GCRB registrant will be raised with the AHCS, which will initiate and manage a Fitness to Practise investigation if it is found to be necessary. The expectation of employers is that, in order to be able to practice, genetic counsellors must be registered with a recognised registration body which, for most genetic counsellors in the UK means the AHCS.The NHS Scientist Training Programme now has a genomic counselling branch leading to statutory registration with the Health and Care Professions Council (HCPC). Here, the legally protected title that genetic counsellors will use is ‘Clinical Scientist’, and, if they so choose, they can suffix this with genetic or genomic counsellor. The AHCS offers an equivalence route for those genetic counsellors who have not had the opportunity to undertake the Scientist Training Programme. HCPC registered genomic counsellors may also apply for AHCS registration through the GCRB if they wish and some have chosen to do so to enhance collective belonging within the profession.The AHCS promotes genetic counsellor registration to ensure protection for the patients and public they serve.Historically, the profession of genetic counselling emerged out of nursing practice. Indeed, in the UK, alongside the Masters training in genetic counselling, there is a specialist nursing route into the profession as well. With the advent of the new Genomic Medicine Alliances in England (GMA) , it is likely that the profession of genetic counselling can be aligned with the Nursing GMA, which will be different from medically trained clinical geneticists colleagues who could naturally align with the Medicine GMA.Classic models of line-management are evolving, such that whilst many genetic counsellors will remain within the specialised clinical genetics services, they may also be commissioned via other funding routes and thus work and are line-managed independently of the traditional clinical geneticist/clinical laboratory scientist/genetic counselling model of service delivery. Commissioning models need to consider the allocation of resources specifically to genetic counsellors working separately to clinical genetics services.As a professional body, the AGNC supports the evolution of the genetic counselling role so that independently practicing genetic counsellors can function, allied to, but working within, services external to the traditional clinical genetics model of care. Genetic counsellors need to be accountable for their own practice and to their own professional regulator as opposed to needing to be ‘responsible to’ clinical geneticists or another mainstream medical colleague. Nowadays, it is more usual for a genetic counsellor to be line-managed and held accountable to a consultant genetic counsellor, rather than a consultant clinical geneticist. For genetic counsellors working in mainstream settings, they may wish to retain formal links with their consultant genetic counsellor colleagues for line-management, mentorship and counselling supervision.As the genetic counsellor is highly skilled in the adaptation of complex information for families, it follows that they have an essential role to play in the delivery of educational materials, resources and teaching for healthcare professionals. Genetic counsellors have long held responsibilities for providing genetic risk estimation training to non-genetic healthcare providers such as breast care nurses based in NHS breast care units or foetal management training to specialist midwives and registrars training in foetal medicine. Training has also extended to GPs.With the implementation of new genomic medicine services across the NHS, hundreds of thousands of new patients will routinely undergo genomic testing as part of their diagnostic pathway. Many genomic results will not only have implications for the patient, but also for their biological relatives too. ‘Care of the family’ and testing of relatives will necessitate a referral of the patient, post result, into a clinical service that can cascade out testing within a family. Genetic counsellors are capable of picking up these referrals and either delivering the genetic counselling themselves or mentoring/line managing other health professionals to do this . Workforce planning and the need for more skilled staff, including genetic counsellors, is recognised as an issue that needs addressing both in the UK and internationally.Urgent modelling is needed to explore how the role of the genetic counsellor can evolve to meet the demands of genomic medicine delivered outside of traditional clinical genetics services. Genetic counsellors are an agile profession capable of playing a pivotal role in supporting the delivery of genomic healthcare both within existing pathways but also within new service delivery.


Kocuria rhizophila exhibited marked and distinctive sensitivity to allosteric activation by GlcN-6P when producing ADP-Glc from glucose-1-phosphate (Glc-1P) and ATP. This behavior is also seen in the enzyme from Rhodococcus spp., the only one known so far to portray this activation. GlcN-6P had a more modest effect on the enzyme from other Actinobacteria (Streptomyces coelicolor), Firmicutes , and Proteobacteria (Agrobacterium tumefaciens) groups. In addition, we studied the catalytic capacity of ADP-Glc PPases from the different sources using GlcN-1P as a substrate when assayed in the presence of their respective allosteric activators. In all cases, the catalytic efficiency of Glc-1P was 1–2 orders of magnitude higher than GlcN-1P, except for the unregulated heterotetrameric protein (GlgC/GgD) from Geobacillus stearothermophilus. The Glc-1P substrate preference is explained using a model of ADP-Glc PPase from A. tumefaciens based on the crystallographic structure of the enzyme from potato tuber. The substrate-binding domain localizes near the N-terminal of an α-helix, which has a partial positive charge, thus favoring the interaction with a hydroxyl rather than a charged primary amine group. Results support the scenario where the ability of ADP-Glc PPases to use GlcN-1P as an alternative occurred during evolution despite the enzyme being selected to use Glc-1P and ATP for α-glucans synthesis. As an associated consequence in such a process, certain bacteria could have improved their ability to metabolize GlcN. The work also provides insights in designing molecular tools for producing oligo and polysaccharides with amino moieties.We explored the ability of ADP-glucose pyrophosphorylase (ADP-Glc PPase) from different bacteria to use glucosamine (GlcN) metabolites as a substrate or allosteric effectors. The enzyme from the actinobacteria  These reactions are, respectively, catalyzed by ADP-Glc pyrophosphorylase  and glycogen synthase (EC 2.4.1.21). The pathway completes the formation of glycogen via the branching enzyme (EC 2.4.1.18), which introduces α-1,6-ramifications  and is allosterically regulated by critical intermediates of the central metabolism in the respective microorganism. For example, fructose-1,6-bisphosphate or fructose-6-phosphate (Fru-6P) and pyruvate (Pyr) are major activators of the enzyme from Escherichia coli (performing glycolysis by the Embden–Meyerhof pathway) and Agrobacterium tumefaciens (using the Entner–Doudoroff route), respectively (glgC and glgD) codify for ADP-Glc PPase, giving rise to tetrameric GlgC or GlgC/GlgD active forms  is the only known ADP-Glc PPase exhibiting insensitivity to allosteric regulators to our knowledge  is the primary allosteric activator    and thatroteins) .Rhodococcus (i) allosteric effectors enhance this activity and (ii) GlcN-6P is a primary allosteric activator  metabolism in actinobacteria of the genus dococcus , 2021. Activator . These fE. coli Top 10 (Invitrogen) was used for plasmid maintenance, and proteins were expressed in E. coli BL21 (DE3) (Invitrogen) using the pET28b vector (Novagen). DNA manipulations, molecular biology techniques, and E. coli cultivation and transformation were performed according to standard protocols  or the highest quality available. rotocols .A. tumefaciens: E187A Forward: CGACTTCATCGCCAAGCCGGC and E187A Reverse: ATGATCTCGTCTTTTTCGTTCACATGC.Mutagenesis was performed by using a Q5 Site-Directed Mutagenesis Kit from New England BioLabs. Oligonucleotides were synthesized by Integrated DNA Technologies . The primers listed were used to make E187A in ADP-Glc PPase from E. coli BL21 (DE3) (Invitrogen) transformed with a recombinant plasmid  harboring the desired glgC gene. Transformed cells were cultured at 37°C and 200 rpm in LB medium  containing the corresponding antibiotics at its proper concentration  until optical density at 600 nm reached between ~0.6 and 0.8. Then, protein expression was induced by adding 0.1–0.4 mM isopropyl-β-D-1-thiogalactopyranoside (depending the protein), which continued for 16 h at 18°C–24°C. Then, recombinant cells were harvested with a 10-min centrifugation at room temperature and 5,000 × g, and the obtained pellets were stored at −20°C until used for purification purposes.Proteins were obtained as previously described , 2020. BBuffer H , disruption by sonication , and then centrifuging twice at 30,000 × g for 10 min. The resulting supernatant (crude extract) was loaded on a His-Trap column  that had been equilibrated with Buffer H. Elution of the recombinant proteins was produced using a linear imidazole gradient (10–300 mM) in the same buffer. Fractions exhibiting the highest ADP-Glc PPase activity were pooled and concentrated. After concentration, the ADP-Glc PPases from Streptomyces coelicolor, Rhodococcus fascians, and R. albus were dialyzed against Buffer S . The resulting purified enzymes were stored at −80°C until use. Under these conditions, enzymes remained active for 6 months (at least).Immobilized metal affinity chromatography (IMAC) at 4°C was the procedure to purify the His-tagged proteins. Protein purification started with cells resuspension in Protein concentration was assayed by the Bradford method , with the use of bovine serum albumin (BSA) as a standard. The purity of the recombinant proteins was assessed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), as described elsewhere . After ri after PPi hydrolysis by inorganic pyrophosphatase, using the colorimetric method described before .The ADP-Glc PPase activity was determined at 37°C in the direction of ADP-Glc synthesis following the formation of Pd before . ReactioOne unit of activity (U) is defined as the amount of enzyme catalyzing the formation of 1 μmol of product per min under the above-specified conditions.Vmax), the Hill coefficient (nH), and the concentrations of activator (A0.5) or substrate (Km), giving 50% of the maximal activation or velocity, respectively. Kinetic constants were reproducible within a range of ±10%, being the mean of three independent sets of data.Saturation curves were constructed by assaying enzyme activity at different concentrations of the variable substrate or effector, while the others remained at saturating levels. Plots of enzyme activity (U/mg) vs. substrate (or effector) concentration (mM) were used to calculate the kinetic constants by fitting the experimental data to a modified Hill equation . Data weA. tumefaciens ADP-Glc PPase structure with ADP-Glc was built using the following templates: the crystal structure of the small subunit of the potato (Solanum tuberosum) tuber in complex with ADP-Glc (PDB ID: 1YP4) and A. tumefaciens ADP-Glc PPase , which was a small effect compared to the 27-fold activation exerted by the major effector Glc-6P (K. rhizophila ADP-Glc PPase responded to the GlcN-6P stimulus to the same extent as to Glc-6P (4–5-fold increase), thus behaving similarly to the rhodococcal enzyme  than their main effector PEP  of the ADP-Glc PPase from two Firmicutes species:  GlcN-6P , even wh GlcN-6P . In contctor PEP , as show–20-fold . Taken tRhodococcus genus and are the only known studies regarding GlcN-1P consumption . The S. coelicolor ADP-Glc PPase poorly used GlcN-1P and only showed a low activation degree in the presence of Glc-6P and GlcN-6P. GlcN-6P barely doubled the specific activity, and while Glc-6P increased activity 4-fold, it remained in the mU/mg order  increased GlcN-1P activity by two orders of magnitude, reaching up to ~20 U/mg. In terms of catalytic efficiency, the K. rhizophila ADP-Glc PPase used GlcN-1P with an efficiency that is almost one order of magnitude lower compared to Glc-1P , we observed 10% activity relative to Glc-1P (not shown), similar to the rhodococcal ADP-Glc PPase   for GlcN-1P. Furthermore, in the presence of 1 mM GlcN-1P, ATP reached similar kcat and Km (0.21 mM) levels  . It is in Glc-1P , sustainn Glc-1P  showing ) levels  with a creported .G. stearothermophilus GlgC/GlgD heterotetrameric structure of ADP-Glc PPase has the highest reported catalytic efficiency to use ATP and GlcN-1P allowing us to hypothesize that it may be metabolically meaningful  for GlcN-1P. The catalytic efficiency for GlcN-1P in the heterotetrameric G. stearothermophilus enzyme falls within the range of a plausible and meaningful metabolic reaction, according to the hypothesis presented elsewhere  exhibiting selectivity ratios with respect to Glc-1P in the range of 50–80  and the α-helix, will contribute to selecting a hydroxyl over an amino group, and as such, prefer Glc-1P over GlcN-1P.The only structural difference between Glc-1P and GlcN-1P is that an amino group is in position 2 rather than a hydroxyl group. Therefore, to understand the selectivity between Glc-1P and GlcN-1P in ADP-Glc PPases, it is important to analyze the structural determinants for the binding of the hydroxyl group in position 2 when Glc-1P is the substrate. Using the elucidated crystallographic structure of ADP-Glc PPase from potato tuber, we modeled the sugar-phosphate binding site in  α-helix . This is α-helix . The hyd α-helix . In addi modeled , yieldedE. coli ADP-Glc PPase and was found to be critical to the binding of Glc-1P  enzyme  enzyme . In the i) bioprospecting of metagenomic data and discovery of new types and families of enzymes or (ii) secondary activities and/or promiscuity of enzymes producing molecules at sub-optimal metabolic concentrations (Pyrophosphorylases (PPases) are critical enzymes for carbohydrate metabolism in different organisms, catalyzing the reversible production of nucleoside-diphospho-sugar and their derivatives. It is described that ~70 nucleotide sugars were identified in bacteria , and thitrations . The fortrations , and hertrations .UDP-Glc and ADP-Glc are synthesized from Glc-1P and their respective nucleoside-triphosphate . In bacteria, the formation of NDP-Glc represents a branching point that determines the fate of Glc in the cell: ADP-Glc is used to form glycogen, while UDP-Glc is shuttled to various metabolic routes . UDP-GlcRhodococcus spp., which exhibits the ability to use GlcN-1P with high efficiency, being the preferred substrate  or an effector (GlcN-6P). We identified these molecules are related to all the characterized enzymes with common patterns and characteristics depending on the source. Recently, several crystallographic structures were obtained for ADP-Glc PPases , and theThe hydroxyl of Glc-1P (or the product ADP-Glc) in position 2 donates a H to the O of the carboxylate from E187 and stands close to a positive density at the N-terminus end of the α-helix F233-H244. This led to the hypothesis that these two major structural elements contribute to selecting a hydroxyl over an amino group, and as such, preferring Glc-1P over GlcN-1P. However, this was not exactly the case. The mutation E187A decreased the overall activity of the enzyme and the apparent affinity for the sugar phosphates, but the selectivity ratio only decreased minimally. This indicates that the H-bond with E187 is a strong feature for the binding of the sugars, but it is not a major contributor in selecting Glc-1P over GlcN-1P. It is well known that at the N-terminus of an α-helix there is an electrostatic positive density that is important for binding of anions . This elRhodococci using GlcN phosphate metabolites led us to propose a putative metabolic node or bifurcation at the level of GlcN-1P, incorporating the characterization of other enzymes, such as GlmU . Worthy of mentioning, the literature available concerning GlcN utilization by microorganisms is scarce and limited to a group   in processes regarded to cell differentiation  , whereasntiation . Accordiodococci . In thisodococci .Kocuria and Rhodococcus. Remarkably, little activation was observed in the R. albus GlgC and GlgC/GlgD enzymes, although it was already reported they had an inherent low sensitivity to effectors for the canonical substrates  is enhanced in the heterotetramer GlgC/GlgD. In this case, the “allosteric behavior” exerted by the GlgD subunit increases the efficiency 60-times for GlcN-1P which is analogous to the “switch-on” mechanism described in the Kocuria/Rhodococcus enzymes. For the canonical substrate Glc-1P, the G. stearothermophilus GlgD subunit increases the catalytic efficiency ~3-fold. Thus, this GlgD behavior resembles the action of the GlgD subunit from Streptococcus mutans on the activity with Glc-1P (Ostreococcus tauri was considered a “specifier” because its presence in the heterotetramer only increases the specificity for the activator 3-phosphoglycerate (3-PGA) compared to fructose-1,6-bisphosphate (G. stearothermophilus (or other Bacilli) remains to be solved; this case also illustrates how enzymological data bring new questions regarding the metabolism while opening the door to novel biotechnological tools.In reported . The genmulation , 2004. Eh Glc-1P . It has hosphate . A classhosphate . The phyE. coli demonstrated its ability to harness the underground metabolism to discover new metabolic functions ((i) new metabolite links (“serine shunt”) were identified, allowing to rewire central metabolism, particularly the Embden–Meyerhof–Parnas glycolysis ((ii) elucidated the appearance of new pathways for isoleucine biosynthesis  when the bacterium grows in GlcN as a sole carbon source (G. stearothermophilus) may provide the potential for the evolution of alternative/novel GlcN-related pathway(s), to yet unknown intracellular fates. Reinforcing this hypothesis, we recently have published the discovery of a new type of pyrophosphorylase specific for GlcN-1P  glycosyltransferases ynthesis . E. colin source , where sn source . However GlcN-1P . Beyond strates) , 2008. Tstrates) . Among tsferases  and incosferases .The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.MA, MB, and AI conceived the study, designed the experiments, and analyzed the results. JB, MI, RM, and AC performed the enzymatic characterizations. JB and MB produced the homology model. JB, MA, MB, and AI wrote the manuscript with contributions from all authors. All authors contributed to the article and approved the submitted version.Cientibeca) from UNL.This work was supported by grants from ANPCyT to MA (PICT-2018-00698), to AI  and from NSF to MB (MCB 1616851). MA and AI are investigators from CONICET. MI is a Fellow (The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.https://www.frontiersin.org/articles/10.3389/fmicb.2022.867384/full#supplementary-materialThe Supplementary Material for this article can be found online at: Click here for additional data file.


Formica ants. Alternative supergene haplotypes are associated with the presence of either a single queen (monogyny) or multiple queens (polygyny) within colonies. Here, we assess the social structure and supergene status of the North American species Formica neoclara.Supergenes are chromosomal regions with tightly linked clusters of alleles that control compound phenotypic traits. Supergenes have been demonstrated to contribute to the maintenance of polymorphisms within populations in traits as diverse as mimetic wing coloration in butterflies, mating strategies in birds, and malarial susceptibility in mosquitoes. A large supergene also underlies variation in social organization in F. neoclara is socially polymorphic in queen number, and we show that the social polymorphism is associated with alternative haplotypes at the social supergene. Intriguingly, polygyne colonies can harbor workers that are homozygous for both haplotypes as well as heterozygotes.We sequenced a subset of the genome in 280 individuals sampled in populations from California to northern British Columbia using ddRADseq. We determined that Formica species, in which almost all individuals in polygyne colonies have the polygyne-associated haplotype. The social polymorphism is present in widely distributed and genetically subdivided populations of F. neoclara. In studying this system in F. neoclara, we expand our understanding of the functional evolution of supergene haplotypes as they diverge in different lineages.This colony genetic composition contrasts with other The online version contains supplementary material available at 10.1186/s12862-022-02001-0. Heliconius butterflies [Stable multilocus genetic polymorphisms often underlie complex phenotypic variation within populations –4. Such terflies . Supergeterflies , includiterflies –16.Solenopsis invicta and Formica selysi (Hymenoptera: Formicidae) [Formica [Solenopsis species [S. invicta, the supergene haplotype found exclusively in polygyne colonies (Sb) selfishly promotes its propagation via a green-beard effect [S. invicta embryos, but this mechanism does not appear to consistently favor one haplotype over the other; instead, supergene-linked loci transmission ratios significantly differed from Mendelian ratios, with some queens producing more Sb eggs than expected and others producing more SB eggs [Formica selysi also favors the polygyne-associated haplotype (Sp) through maternal effect killing [Independent and distinct supergenes that underlie a polymorphism in colony queen number were initially described in two ant species, micidae) , 14. Monmicidae) –19. Thes[Formica , 20 and  species , meaningd effect , in whicd effect . Transmi SB eggs . The sel killing , whereinSolenopsis and Formica species in the distribution of supergene genotypes in colonies. In both cases, monogyne colonies contain exclusively one supergene haplotype, SB in S. invicta and Sm in F. selysi [S. invicta colonies possess SB/SB and SB/Sb workers and SB/Sb queens. Sb/Sb females rarely reach adulthood [F. selysi colonies do not contain Sm/Sm homozygotes (workers or queens), but they do have Sp/Sp and Sm/Sp workers and queens [Formica species [There are some notable differences between . selysi , 14. In dulthood , 24, potdulthood . Polygynd queens . We are  species , 20. UndFormica neoclara is an ant species found throughout western North America. Workers forage in trees [F. neoclara [F. neoclara remain largely unknown.in trees , where tin trees . Past rein trees  and agrineoclara , 26. DesF. neoclara populations distributed from California to Northern British Columbia to determine whether the species is socially polymorphic and, if so, whether colony queen number is under genetic control throughout its range. Further, we investigate the genetic structure of populations across the range of this species to determine whether the population likely expanded recently or whether geographically distant populations are also genetically distant. Overall, this study will begin to uncover similarities and differences in social polymorphisms and their genetic bases in ant species with distinct evolutionary histories.Here, we investigate Using multiple complementary metrics based on worker genotypes (based only on loci outside of chromosome 3), we determined the colony queen number and estimated mate number for 28 out of 32 colonies  are heterozygous (Sm/Sp), and individuals in the right cluster (yellow) are homozygous for the alternative haplotype (Sp). Looking across PC2 and PC3, we observe signatures of geographic variation within each supergene genotype cluster . These SNPs lie between 7.7 Mb and 12.6 Mb  to identify single nucleotide polymorphisms (SNPs) associated with variation in colony queen number. When we restrict the analysis to the socially polymorphic Alberta population , we see a strong association between five SNPs on chromosome 3 and colony social form . In contrast, all but one colony determined to be monogyne are composed of Sm/Sm workers exclusively. In one exception, the solitary colony from Idaho (PocC4) contains both Sm/Sm and Sm/Sp workers, and we deemed this colony to have one multiply-mated queen on the basis of the low opposing homozygosity and high relatedness values . This phenotype call could be an error; we expect relatedness estimates to be inflated when local samples sizes are small. In parallel, workers from PocC4 exhibited an elevated FF. neoclara colonies contrasts with previously studied ant species with social supergenes .Expected heterozygosity values from Alberta (sites from the plains and Rocky mountains grouped separately in the map), northern and southern British Columbia, California, and Idaho range from 0.15 to 0.27, with an average of 0.235  [F. neoclara polygyne colonies can harbor individuals lacking the Sp allele, with some colonies containing all three possible genotypes . Out of 18 polygyne colonies sampled, we never detected a multiple queen colony with exclusively Sm/Sm individuals. This pattern suggests that the association between the supergene and colony queen number is present in F. neoclara, as in other Formica species, despite differences in haplotype distribution within colonies. species  is also es Figs.  and 4. Id Sp/Sp) , 19. In F. neoclara. Finding Sm/Sm workers in polygyne nests in all populations suggests that the ‘maternal effect killing’ selfish genetic mechanism found in F. selysi is not operating in F. neoclara [F. selysi clade or whether it was lost from the F. neoclara clade. Preliminary evidence from a small number of colonies suggests that F. cinerea, a close relative of F. selysi, may have Sm/Sm genotypes in polygynous colonies in some populations [F. glacialis, a relative of F. neoclara, detected no Sm/Sm workers [F. selysi colonies at both local [S. invicta and F. selysi appear to be weak or absent, further research is needed to determine what factors maintain this genetic polymorphism in F. neoclara.The distribution of genotypes within polygyne colonies raises questions about how the genetic and phenotypic polymorphisms are maintained in neoclara . Based oulations . A simil workers . Studies workers , 29, 30.th local  and regith local . HoweverFormica species, with Knockout standing out as a strong candidate gene [Knockout or in other candidate genes identified by previous studies [GEMMA analysis of all populations reveals that at least one SNP on chromosome 3 (12.1 Mbp) is associated with social form. Restricting the analysis to colonies from Alberta shows that five SNPs on chromosome 3 are correlated with social form. A handful of genes associated with or around regions of the supergene (chromosome 3) are conserved across multiple ate gene , 20. NonTo assess colony social structure, we employed a method that evaluates the opposing homozygosity of biallelic RAD loci and the nestmate relatedness in parallel Fig. . MembersWe first note that both opposing homozygosity and relatedness result in some ambiguity at intermediate values. Specifically, intermediate levels of opposing homozygosity and average relatedness can be found in two types of colonies: those containing a polyandrous single queen or a small number of related nestmate queens (i.e. oligogynous colonies). We assessed the distribution of relatedness values in colonies determined to be monogyne, undetermined, and polygyne from the same population , 70% ethanol for the second DNA wash, and eluted the DNA in 30 µL of buffer AE. We then used a double-digest restriction site associated DNA sequencing (RADseq) approach to sequence samples . BrieflPEAR [Formica selysi reference genome [BWA and called genetic variants across the sample using BCFtools mpileup [We demultiplexed reads across each of the three batches using the process_radtags (version 1.4) command in Stacks, with default parameters . To mergPEAR . We thene genome  using BW mpileup .VCFtools (v 0.1.13) [--minDP 7), to remove loci that were present in fewer than 80% of samples (--max-missing 0.8), and to remove sites with a minor allele frequency less than 0.05 (--maf 0.05). Samples with more than 20% missing data were removed prior to analysis. Batch effects were evident with more permissive max-missing thresholds, but the threshold of 80% ensured that retained loci were present in all three sequencing batches   for missCOLONY program [COLONY once for each region. We excluded colonies with fewer than six workers  from colony-level analyses. After inferring queen number, we estimated the average relatedness among workers for the 32 colonies using several estimators. Relatedness calculations include the Ajk statistic (--relatedness) [VCFtools as well as the Huang diploid A estimator available on the PolyRelatedness program (e 14 0) [We assessed colony composition using multiple metrics, allowing us to come to a consensus to infer colony queen number. To ensure that these analyses were independent of our assessments of supergene variation, these analyses excluded all markers on chromosome three. The  program  allowed tedness)  and kinstedness)  available 14 0) . The una(e 14 0) . KING us(e 14 0) . The Hua(e 14 0) . Therefo(e 14 0) , which c4 0 . The(e 14 0) . Finally(e 14 0) ). We calF. selysi [plink  [IS, value above zero were homozygous, while those with a FIS value below zero were heterozygous . Individuals within the center cluster on the PCA  of markers from chromosome 3 Fig. . The kno. selysi , was anatra-chr)  (n = 26 Beagle (v 5.1) [F. neoclara genetic data. GEMMA [-k -lmm 1). We then visualized output data from this process via a Manhattan plot  analysis to test for an association between each locus and the inferred social form of each colony. This GWAS was performed independently of genotype assignment, using only social form assignment and markers on chromosome 3. We ran two analyses: one for colony samples from all regions  and one for colonies from Alberta, Canada only . The latter analysis reduced the effects of population structure on the analysis. We excluded workers from transect samples and colonies labeled as ambiguous in social form from these analyses .  (v 5.1)  was useda. GEMMA  was usedVCFtools) for each population as the average nucleotide diversity per variable site on all chromosomes except chromosome 3 of one individual per colony  and all transect samples. Following this, we performed a PCA using all markers except those on chromosome 3 . Lastly, we performed a pairwise isolation by distance (IBD) analysis on the 32 colonies. Like previous colony-level analyses, we excluded colonies with fewer than six individuals. We calculated the Weir and Cockerham FST [VCFtools. We calculated the distance between colonies using the Imap package (v1.32) [To observe whether the geographically distant populations show signs of historic isolation or recent expansion, we utilized SNPs not on chromosome 3 for multiple analyses. We calculated expected heterozygosity at variable sites   in R [52 (v1.32)  genetic Additional file 1: FigureS1.A Results of GEMMA analysis utilizing workers from colonies fromall regions, visualized via Manhattan plot. We used a linear mixed model withcolony social form as the independent variable. Each point represents anindividual SNP, with the corresponding chromosome on the x-axis and thenegative logarithm of the SNP p-value on the y-axis. Only one SNP, from chromosome 3, exceeds the significance level . B Results of GEMMA analysis onworkers from colonies in Alberta only, to reduce the effect of underlyingpopulation structure on the GWAS, visualized via Manhattan plot. We used alinear mixed model with colony social form as the independent variable. Eachpoint represents an individual SNP, with the corresponding chromosome on thex-axis and the negative logarithm of the SNP p-value on the y-axis. Five SNPs,all from chromosome 3, are above the significance threshold . Figure S2. Stacked bar plot displaying genotypes of samples from colonies labeled as ambiguous in social form . Each bar represents all samples from an individual colony. Genotype in relation to each individual worker, is indicated by color: green = Sm/Sm andbrown = Sm/Sp. Figure S3. Line graph showing the effects of missingness on the Huang estimator. Individual (red) and whole sample (black) relatedness values are represented as lines. Figure S4. Density plot showing pairwise relatedness (determined by PolyRelatedness) among all sequencedworkers of colonies from the Evan Thomas trailhead in Alberta, Canada. On average, workers from monogyne colonies are the most related, whereas intermediate colonies (“ambiguous” in Fig. Figure S5. A principal component analysis utilizing all markers except those on chromosome 3 in individuals fromcolony samples, with the year of the respective batches color coded. Table S1. Overview of consensus social form variables. Table S2. Sampling regions and their associated coordinate values. Table S3. PC axis weightings of variants on chromosome 3 (see also Fig.


Genetic counselling has been defined as the process of helping people “understand and adapt to medical, psychosocial, and familial aspects of genetic conditions.” It can also help patients and families deal with stigma and understand the significance of possible genetic findings. Psychiatric genetic counselling (PGC) is an emerging field aimed to help people with a personal or family history of psychiatric illnesses such as schizophrenia, bipolar disorder, or neuropsychiatric conditions, to understand genetic etiological mechanisms as a critical component. Counselling strategies are used to identify and adapt to psychological and familial consequences of the conditions and to reduce stigma surrounding the psychiatric illness. A recent survey showed that PGC is still not routinely offered and usually only discussed at the initiative of the patient, e.g. if they ask about the possibility of “hereditary" illness, or if a caregiver during a session for another indication, identifies the family history. If a monogenetic or chromosomal cause is identified, the genetic counselling follows a more traditional path, but if, on the other hand, the cause is complex, the counselling will not be as clearcut. It will then focus on explaining risk for disease with quite uncertain riskscores as no causative genetic change is identified. Although genetic testing most often cannot be offered and individual risk scores based on genetic markers cannot be given, there is still great value for patients and their relatives in PGC. Studies have shown that the effect of PGC is an increase of empowerment and a reduction of stigma.No significant relationships.