Lactobacillus, Bacillus, and Saccharomyces. We identified a major gap of research on uncommon beverages and poor attention on the cultural and technological aspects. TMFB are dynamic and heterogenous foodscapes that are valuable biocultural reservoirs. Policies should include their promotion for conservation. The main needs of research and policies are discussed.Mexico is one of the main regions of the world where the domestication of numerous edible plant species originated. Its cuisine is considered an Intangible Cultural Heritage of Humanity and ferments are important components but have been poorly studied. Traditional fermented foods are still diverse, but some are endangered, requiring actions to promote their preservation. Our study aimed to (1) systematize information on the diversity and cultural history of traditional Mexican fermented beverages (TMFB), (2) document their spatial distribution, and (3) identify the main research trends and topics needed for their conservation and recovery. We reviewed information and constructed a database with biocultural information about TMFB prepared and consumed in Mexico, and we analyzed the information through network approaches and mapped it. We identified 16 TMFB and 143 plant species involved in their production, species of Cactaceae, Asparagaceae, and Poaceae being the most common substrates. Microbiological research has been directed to the potential biotechnological applications of  The consumption of ferments by humans probably started unintentionally since fermentation occurs in nature as a common process . HoweverCurrently, a small fraction of species of organisms are predominantly used in the human diet worldwide, and this fact makes the hegemonic food systems vulnerable . FermentFermented products are outputs of complex interactions of three main components, substrates, microorganisms causing fermentation, and humans who drive the process and value the final products ,12. HumaThe fermented foods and beverages were used long before any awareness of microbiology and biochemistry , but nowFermented beverages are among the most iconic fermented foods worldwide; these are essential components of local diets in many cultures and are mainly prepared from plant substrates ,25,26,27The traditional ecological knowledge involved in the preparation of fermented products appears to be at risk because of the overreliance on the massive campaigns of commercially produced beverages, which reach rural regions where the traditional products are prepared ,30. ThisThe human diet worldwide is vulnerable since it is mostly limited to 10 to 50 plant species that provide about 95% of the world’s caloric intake ,33,34,35In 2010, the UNESCO recognized food as an intangible cultural heritage and included the Mexican gastronomy in the representative list of “Intangible Cultural Heritage of Humanity” ,42. HoweIn Mexico, a broad spectrum of traditional fermented foods, including beverages, is produced from different raw materials by Indigenous and mestizo people in different regions, the processing methods and microbiota varying among regions, localities, and producers ,48,49,50Several studies have been directed to characterize the microbial communities associated with some TMFB ,48,49,50mescal [pulque [pozol [The characterization of microbial communities of Mexican fermented products has been commonly performed by culture-dependent approaches. Early classifications of products were conducted based on the physiological and biochemical criteria and microscopic inspection . Nowadaymescal ,48, pulq [pulque , and poze [pozol . Althougpulque [atole agrio [tuba [More recently, some studies have used high throughput sequencing (HTS) techniques, such as Illumina MiSeq paired-end sequencing of barcoded polymerase chain reaction (PCR) amplicons, but these studies are still scarce in TMFB. However, studies with shotgun metagenomics have recently been conducted to characterize and infer the dynamics of microbial communities of pulque ,57, atolle agrio , and tubio [tuba . A few sio [tuba .This review started with the assumption that the TMFB are the outcome of the synthesis of the vast biological and cultural diversity. In particular, we expected to identify a high diversity of TMFB, which is related to the similarly high cultural diversity represented by the number of Indigenous groups that are distributed throughout the different regions and ecosystems of Mexico. We expected that those more widely distributed and more studied groups from ethnobiological perspectives  are alstepache, pulque, mescal, colonche, jobo or hobo, colonche, nawait, pozol, tejuino, tesgüino, piznate, taberna, cocoyol, tuba, Mexican palm wine, balché, xtabentún. As these names are mainly in Spanish, we used the Boolean operators OR and AND. For example, in the Web of Science, the following search string was used: “Agave” AND “fermentation”, “tradicional” AND “bebidas”, “traditional” OR “ancestral” to improve the research parameters. We extended the search in local databases using keywords such as Mexican fermented beverages, bebidas fermentadas mexicanas, fermented beverages in Mexico, bebidas fermentadas en México, fermented products in Mexico, productos fermentados en México, tepache, pulque, colonche, hobo, Mexican wines, vinos mexicanos, colonche, nawait, pozol, sidra, tejuino, tesgüino, taberna, cocoyol, tuba, Mexican palm wine, and balché.We conducted a search of peer-reviewed literature in Scopus, Google Scholar, Google, and the Web of Science databases to visualize the current state of research on TMFB. In addition, since numerous studies are not covered in these databases, we reviewed local journals, technical reports, books, and Ph.D. theses from regional universities of Mexico to complement the information. We also consulted references from gastronomic literature and governmental sources information. Through this search, we identified 328 peer-reviewed articles plus 197 other references related with TMFB. A search of peer-reviewed literature was performed with the following keywords: Mexican fermented beverages, traditional fermented beverages, pulque mixed with several fruits known as pulques curados; in this case, we only considered the name pulque for all of these beverages. In a broad sense, in this review, we only considered the main substrates for fermentation and secondary substrates employed during the fermentation process.Most of the beverages recorded are wine-like beverages produced by the fermentation of several fruit species; however, we found numerous cases in which the producers use to add sugar cane alcohol or spirit beverages to confer specific flavors, and these beverages were not considered in our analysis. We found different names for beverages produced with To characterize the current state of research on TMFB, the information of the title and the abstract of each article were used as inputs in the software VOSviewer 1.6.15 ,63. The The information from this search was systematically stored in a database using Access with the following fields: (1) Fermented product name; (2) plant substrates  microorganisms ; (4) ethnic groups producing and using the beverage ; (5) sensorial features associated to beverages ; and (6) medicinal . Searches in the databases considered studies from 1960 to August 2020. In addition, to document the conservation policies over biological resources and traditional knowledge, we searched in governmental agencies databases.A map was constructed to visualize the documented distribution of fermented beverages based on the data collected in the literature; the area of the municipalities identified was considered as the documented distribution area. Another map was constructed based on the estimated distribution regions proposed by Bruman . In addiTo visualize the uniqueness, diversity, and interaction of the substrates employed to produce fermented beverages, and the co-occurrence of specialist or generalist species in microbial communities, we performed an exploration through the bipartite network approach. The network’s theory offers powerful tools to describe complex communities and the distribution of species specificity within them. Moreover, this approach was performed with the microorganisms identified in the literature to each traditional fermented beverage. The data from the network structure were used as descriptors of diversity and uniqueness. The analysis was performed through the Rstudio software  with theAgave plants and the addition of fruits of native and introduced plant species. By the current review, we identified 16 names for traditional fermented beverages produced from several substrates such as seeds, stems, fruits, tree barks, fruit pulps, and sap. Stems are the dominant substrates used for preparing traditional fermented beverages. We identified 140 plant species used as main substrates for fermentation or as promoters of fermentation.Based on a previous review performed by Godoy and collaborators , 66 typemescal, pulque, tejuino, pozol, chorote, colonche, saká, sendechó, balché, atole agrio, pox, sambudia, tesgüino, tepache, tuba, and taberna  (in green) is the mescal cluster, in which Agave potatorum appears as the most studied species, and Saccharomyces cerevisiae as the most commonly characterized microorganism. This cluster highlights the economic importance of mescal production, and the interest in improving the fermentation profile using several S. cerevisiae strains. Cluster (3), the yeast cluster (in blue), emphasizes the use of non-Saccharomyces yeast species to improve the aroma profile and optimize the ethanol production in mescal. Cluster (4) is the pathogens cluster (in olive) and refers to the effects of pathogens such as Scyphophorus acupunctatus, which mainly affect the Agave groups, decreasing the production of beverages related to these plants. Cluster (5) is the Agave salmiana cluster (in purple), which is mainly related to the influence of environmental factors in the development of plants and the fermentation process.The current state of research in peer-reviewed articles on TMFB can be visualized in the network of Agave species, mainly those involved in the production of pulque and the implications of management of agaves for sap extraction. Cluster (7), the one of traditional fermented beverages (in orange), features the less common traditional fermented beverages prepared in central Mexico, those with Agave species such as pulque or mescal that occur mainly in central Mexico. Cluster (8), the maize group (in brown), mainly accounts for research directed to isolate and characterize the bacteria and yeasts associated to possible probiotic attributes in tejuino and pozol. Cluster (9), the cultural cluster (in light purple), groups studies related to traditions and tourism, mainly in the state of Oaxaca. It can be highlighted that this cluster is also related to mescal production.The following clusters are related to the most traditional beverages and human groups related to them: Cluster (6), the management cluster (in light blue), groups the management practices on mescal production process and its optimization. This is not surprising because of the national and international rising market boom of mescal. However, it is lower than research on tequila, which is the fourth largest export product of Mexico [mescal, pulque is the most studied beverage, particularly biotechnological and management aspects involved in its production.The network shows that most of the current research is directed to study the f Mexico . After mS. cerevisiae, which is a common species in fermented products around the world and it is frequently found in alcoholic beverages. Surprisingly, it is possible to see in the network that the non-Saccharomyces species form a node; this is a trend around several industrialized beverages, such as beer [The main microorganism referred to in the studies is  as beer , wine [1 as beer , and rec as beer , which l as beer .pozol, pulque, tepache and tejuino, whose microbiota and potential benefits to health have been majorly characterized.Two major trends can be identified in the current research on traditional fermented beverages in Mexico. The first one includes biotechnological approaches, which could be visualized in the first three clusters referred to above, which is related to the worldwide interest of the dairy industry to promote products with probiotic and prebiotic compounds . This toAgave beverages such as pulque and mescal, but also beverages such as pozol, which entails biotechnological approaches and the traditional management of sources of substrates, particularly Agave species. The rising market of mescal appears to be leading this trend, while in the case of pulque, the possible food functionalities appear to be the main drivers of the current research programs. It is possible to see the great cultural node related to mescal production, but few studies focused on the cultural groups in other beverages.The second research trend is related to the Agave products have but almost null on the rest of the traditional fermented beverages. As mentioned, there is high interest in biotechnological research programs specially to explore the central region of Mexico, which could be seen in Figure 4A. The remaining beverages are not only marginalized in markets, but also in the scientific agendas. Greater efforts to maintain them should be performed.There is a huge gap in the research agendas on traditional fermented beverages. It is notorious the major interest that Z. mays (Z), A. salmiana (A), S. officinarum (S), and Cinnamomum verum (C) can be seen involved in the production of different fermented beverages. S. officinarum is a species frequently employed to strengthen fermentation as it is used as an external supply of sugars, commonly added as processed brown sugar. C. verum is used to add different flavors to the final product but not during the fermentation process. Cinnamon has been recorded to have antimicrobial and pathogens inhibitors properties, but it has not been reported on its use to avoid spoilage. Sugarcane and cinnamon are non-native species from Mexico but are strongly integrated into the traditional fermented beverages production as enhancers or additives.We identified 143 plant species used as main and secondary substrates for fermentation, as promoters of the fermentations process, and as additives to improve the shelf life or flavor see . The netThe network of substrates of the traditional fermented beverages of Mexico is diverse (S = 5.21), with a minimal connectance, which indicates the uniqueness of most of the beverages with a high specificity in the substrates. The documentation and protection of those unique substrates should be attended to maintain and guarantee their production. The network allows for visualizing that maize is the most generalist substrate, which is not strange since maize is a keystone of the Mexican food systems and cultures and is prepared in great diverse ways.atole agrio, saká, tejuino, tesgüino, sendechó, chorote, and pozol. Pox was usually prepared with maize stems as the main substrate, but sugarcane stem is more commonly used now. For processing most of these products, people employ specific races of maize; we identified 21 races used to produce them, and the beverages receive different names according to the maize race and locality where they are produced. Beverages produced with maize have a deeply and strong relationship with ethnic groups. Through this review, we identified that maize beverages are the most consumed by 21 cultural groups of Mexico.Eight TMFB involve the use of maize as a main or secondary substrate, most of them are prepared with maize grains, including: Tesgüino is produced and consumed mainly by the Rarámuri or Tarahumara people and other cultural groups such as the Yaqui and the Wixarika, who live in northern Mexico [Stevia serrata, Chimaphila maculata, Datura meteloides, Hamaemelum nobile, and Cojoba arborea, all of them identified with local names [Usnea dillenius, a lichen, is used as a catalyst [n Mexico ,88,89,90al names ,92. Usnetejuino and tesgüino are the same beverage since the fermentation core substrate in both cases is the grounded kernels of maize (a dough); however, the process, the plant species, and the varieties added, locations, and the cultural groups are different. Tejuino is a TMFB prepared with the dough of one variety of maize and the addition of brown sugar. It is mostly consumed in the states of Colima and Jalisco by mestizo people, and non-special containers are used for fermentation. The atole agrio is a similar beverage. It is the fermentation of maize dough, and no sugar is added. It has been recorded in southern Mexico and its consumption involves the use of different local races of maize among the localities.It is commonly assumed that Pozol and saká, are non-alcoholic fermented beverages from the Mayan region, prepared with dry corn kernels, boiled with calcareous stones and then grinded into a dough. This is a process of nixtamalization that confers a particular consistency and flavor to the dough and adds nutrients such as calcium oxide and others that become bioavailable [pozol is more common among the mestizo communities and saká is associated to Indigenous communities in religious contexts. For its production, maize dough is stored into small balls in banana leaves until fermentation, a process that varies depending on the environmental conditions and the producers’ preferences. The dough balls are dissolved in water and consumed as a daily beverage [pozol dough can be mixed with batata, toasted cocoa, coconut, cinnamon, or vanilla [vailable ,81,151. beverage ,92,151.  vanilla ,74,75,76 vanilla ,78,79.Sendechó is a beverage that can be prepared with several maize races (five reported), depending on the producers’ preferences, so its final color might vary from light red to purple, light yellow, white, and black. Through the germinating grains step, Buddleja americana is mixed with the grains [Pulque, called ixquini in the region, is added as a starter inoculum [Agave sap or fermented sap can be added as a starter of fermentation. It is produced in areas close to Mexico City and is still consumed by the Mazahua, Otomi, and mestizo groups for several cultural purposes.e grains ,49. Pulqinoculum ,138. TheAgave beverages is a highly diverse group, with 68 agave species involved in the fermentation of pulque and mescal as the main substrate to produce both beverages. The fermented sap of A. salmiana is the most frequently employed to produce pulque and it is also added as an inoculum to other beverages. Although Agave is a diverse group, we recorded low connectivity among its components, which suggests unique fermented products. Since species and varieties of Agave are not explicitly recorded in some studies, we might be underestimating their distinctiveness; there are several local names for the local varieties, suggesting high variation, but unfortunately, their taxonomic identity has been insufficiently studied and indicated.The cluster of Mescal appears to be the predominant beverage of Agave throughout the Mexican territory. It is in fact the beverage that employs a major number of species to get a final product, most of them are locally distributed and different species are often mixed to produce a unique sensorial profile. We consider mescal as a TMFB because most of the species employed for its production are endemic to Mexico, and most of them wild but some have gone through a domestication process or are under different levels of domestication by people of this country. Moreover, because these products are locally and/or regionally produced, their production is mostly unique for each producer, are part of the local gastronomic heritage, and include local and foreign technologies as mentioned before [Agave stems and foliar bases whose products are then distilled; however, the species used are numerous and variable among the regions.d before ,46,47,48pulque is produced. Moreover, we did not consider the species that are used as tools, such as Lagenaria siceraria, to store or transport the sap of Agave species. However, the number of Agave species employed as main substrates for pulque production (42 species) is high. Pulque results from the fermentation of Agave sap, and it has been recorded as spontaneous fermentation [Pulque preparation starts with the collection of the agave sap called aguamiel directly from the agave stem, which is mostly performed under non-sterile conditions [pulque because of the plethora of practices, species, and conditions for its production. These practices might change the fermentation process in time, the assemblages of yeasts and bacteria, and the final sensorial attributes.As mentioned, we did not consider other plant or animal products that are often mixed with pulque consumption which are generally called pulques curados ,94,95. Wentation ,92, but entation ,94,95,96Pulque has been used historically for festivities, ceremonies, agricultural rituals, births, and funerals [pulque in Mexican law (NMX-V-022.1972), which defines quality standards and sensorial attributes. In addition to the health benefits of pulque or its biotechnological applications, the diversity of plants employed and the practices performed reflect a historical know-how developed by humans in interaction with their local environments. However, this production system has been marginalized.funerals . There iColonche is a traditional fermented beverage that can be prepared with fruits of at least 17 cacti species. Colonche is the common name of this beverage in the region of the Altiplano central region of Mexico, where it is mainly prepared with several Opuntia species, although the most common and preferred by the producers and consumers is that prepared with O. streptacantha [Nawait is another common name for a fermented beverage mainly produced by the fermentation of Carnegiea gigantea fruits, whose distribution covers the Sonoran Desert in northern Mexico and the southwestern U.S.A. [colonche-like beverage is also prepared with Pachycereus pringlei. In the Tehuacán-Cuicatlán Valley in south-central Mexico, colonche is known as nochoctli or pulque rojo. There, the fruits of several cacti are employed to produce this fermented beverage, the most common are those of Pachycereus weberi, Escontria chiotilla, and Stenocereus spp. This beverage is produced in different seasons depending on the availability of the substrates. For instance, in south-central Mexico, colonche is produced in April–May and in August–September, when fruits of the different species of columnar cacti and Opuntia used to produce it are available. The production and fermentations practices of colonche are variable in the different regions [A. salmiana or by using inoculums from previous fermentations of cacti fruits. Most of the fermentations occur in clay pots continually used and that have been maintained by several generations of the managers [tacantha ,50. Nawan U.S.A. . In this regions . In mostmanagers .tuba and taberna form the palm cluster. These are prepared by the fermentation of the sap extracted from different palm trees, similarly as it is carried out with other palm wines consumed around the world, such as legmi in Africa, or kallu in southern India, as well as several Asian palm wines [taberna is Acrocomia aculeata known as coyol, and it is produced in localities of the southern state of Chiapas [tuba is obtained from the inflorescences of Cocos nucifera, which is collected and then stored in plastic containers, although the iconic containers are those manufactured with L. siceraria fruits, which are called bules [The beverages lm wines ,99. The lm wines . These b Chiapas ,154,155.ed bules ,154,155.Balché is an exceptionally important TMFB since it is the sacred drink for the Mayan, consumed in several ceremonies. It is the result of the fermentation of bark from several Lonchocarpus species, among them L. punctatus or L. violaceus, and L. longistylus, which is mixed with honey (either from Apis mellifera or melliponini bees). It has a particular pink color and sweet taste, and has been consumed since pre-Hispanic times [S. cerevisiae has been the only microorganism recorded in the fermentation of this beverage [ic times ,131,132.beverage , but morTepache can be prepared with at least 10 plant species as a substrate. Nowadays, in central Mexico the most common substrate is pineapple (Ananas comosus), but its etymology in the Náhuatl language derives from tepitl, a beverage made with maize in the past and in few localities [tepache is also produced by the fermentation of other fruits that belong to the Bromeliaceae family, such as Bromelia karatas, known as tumbiriche or timbiriche. It can also be prepared with fruits of plants introduced into Mexico such as apple, orange, and guava [tepache prepared with pineapple, the process starts by peeling the infrutescences and adding the rind into a wooden container known as tepacheras, then brown sugar is added. It is a spontaneous fermentation where the microbiota is mainly associated with the sorosis epidermis [tepache with other fruits can be fermented in plastic containers; this beverage is commonly homemade, and its quality varies from kitchen to kitchen.calities ,50, whicnd guava ,118. In pidermis ,50. ProdSambudia is a beverage prepared with several substrates, including maize, rice, and barley. Sambudia is the name used for different beverages produced in the state of Mexico; a first way to produce it is using pineapple peel as a substrate [tepache. A second way to prepare it is with ground grains of rice, or barley; in this case, cinnamon, cloves, pepper, roasted and ground maize leaves, and pulque are added in a container; brown sugar is added to sweeten the mixed elements and the mixture is fermented for at least one day [B. karatas fruits by adding pineapple peel, maize leaf, and ground maize. Fermentation occurs by inoculating the remnants of past fermentations in the containers, which are clay pots called sambudieras [ubstrate ,135,136 budieras .The historical records indicate that practices and techniques for manufacturing fermented foods evolved independently in every hemisphere and were developed based on the resources available in the local environments . In Mexipox prepared with the stems of Z. mays nowadays, mainly prepared with S. officinarum stems [C. verum, also originated in Asia [As humans migrated from region to region, food cultures and production practices moved as well ,104,105.um stems , but alsum stems . C. veru in Asia ,108,109,taberna and tuba are examples of the adoption of new technologies and instruments, including distillers for the Philippine coconut spirits distillation technique [mescal and other beverages [The fermentation of numerous palms sap is common in countries of Asia and Africa ,110,111 echnique ,113,114.everages .The most common practice to promote fermentation worldwide is the inoculation of a substrate with starter cultures. For instance, in Korea, soy sauce is prepared using meju (solely fermented soybeans), whereas in Japan and China it is prepared using koji  . Similarmescal [pulque [atole agrio [Through this review, we identified names such as tibicos, castaña, xinaiste, jinaiste, zinaiste, el pie, el pie de pulque, asiento, ixquini, semilla, xaxtle, and nancle among the most frequent names that fermenters give to those mixed starter cultures for traditional fermented beverages. The study and characterization of these microbial communities have only been scarcely covered in studies of beverages such as mescal ,165, pul [pulque ,166,167,le agrio . This teA commonly accepted assumption in traditional fermented products is that spontaneous fermentations have inconsistent or heterogeneous quality outcomes. However, fermentation managers procure to simplify the diversity of the environmental conditions and practices to decrease such heterogeneity. The practice of selecting fermentative environments or controlled facilities is common; for instance, people procure fermenting inside the house or special areas to control the external environmental heterogeneity in light incidence, temperature, external contaminants, or pathogens. Moreover, there is a cultural selection of the person who performs the fermentation and the substrate’s quality ,163,164.tesgüino [tesgüino clay pots, fermenters use the specific name of tesgüineras and if the clay pot is broken, the pieces are trembled inside a new one, thus maintaining the microbiota from the old one. Nevertheless, few studies have attended to the importance of these containers as reservoirs of microbiota and their relevance in the fermentation process.The interrelationships between the native microbial strains, substrates, techniques, tools, and fermentative environments of TMFB have been little studied. We identified studies that characterized the clay pots or tinas de fermentación for pulque  and tesgtesgüino . In factS. cerevisiae, as the main microorganism responsible for alcoholic fermentation. These are invariably present in mescal and pulque. Nevertheless, for maize fermented beverages, the isolation and characterization of microorganisms have been directed to bacteria to characterize the possible probiotic functions of lactic acid bacteria (LAB) and, more recently, their function as a source of nitrogen fixation bacteria [As mentioned, most studies on microbial environments in fermented beverages have been conducted with culture-dependent methods, isolating the most common microorganisms, among them bacteria .Pozol (Poz), pulque (Pul), and mescal (Mes) are the beverages with more species recorded (Lactobacillus (Lac), Bacillus (Bac), and Saccharomyces (Sac) are shared among most groups of beverages. The network values are shown in We found that only 10 beverages have been studied to characterize their microorganism communities, identifying 255 species of microorganisms. recorded . This corecorded . The netrecorded . In addimescal, pulque, atole agrio, and pozol are the beverages with the highest number of microorganism species identified, while colonche is the beverage with fewer genera identified. In general, six modules of microorganisms in TMFB were detected: (1) mescal; (2) pulque; (3) atole agrio; (4) pozol; (5) colonche, and (6) palm and tepache. This analysis shows that 99 species occur in the most studied fermented beverages, which might be the generalist core of microorganisms intervening in these fermented products. Interestingly, with this analysis, we identified that pozol has 24, mescal 23, pulque 22, atole agrio 10, and colonche 2 genera with low shared similarities with other beverages, and those could be specialist genera in each traditional beverage. Nevertheless, as the production practices and substrates vary for each event of production and ferment type, further analyses are needed for more precise information.Through the nested analysis, we can see that S. cerevisiae in the fermentation of several products [Forthcoming technologies such as CRISPR/cas9 ,173,174 products ,173. NevMestizo people produce and consume most of the TMFB, mainly those prepared with maize and agave as the main substrates. This could be explained because mestizo people are the most numerous in Mexican society and have recovered Indigenous traditions; also, because maize plays the key role in Mexican gastronomy, it is widespread, and fundamental in the local communities’ nutrition ,176. TheIndigenous cultural groups are interested in looking forward and renewing their connections with their lands and cultural heritage and recovering their traditional foodways to regain cultural strength and personal and community health ,179. ThiIt is important to highlight the marked level of segregation of the Mexican Indigenous people, which has been verified by numerous anthropological and socio-demographic studies based on synthetic demographic and poverty indexes ,184,185.Efforts should be directed to characterize and document aspects of TMFB, such as their uniqueness, human cultural specificity, their heritage status, the dietary patterns, their role in cultural identity, practices in the agricultural production of raw materials for fermentation, dishes and gastronomic innovation, preparation techniques, recipes, food traditions, symbolic dimensions, and material aspects such as utensils and dishware, among other biocultural topics. In addition, promotion activities would be relevant. For instance, promoting events in the communities, sponsored through schools, councils, and cultural centers, can be effective to illustrate, highlight, and demonstrate the values of traditional food systems ,187. Thetepache, sendechó, sambudia, atole agrio, saká, tuba, and pozol distribution.We compared the distribution of TMFB documented by Bruman in 1993 B with inbingarrote or bingui from central Mexico, reported by Pineda [binguí or benjuí and the spirit bingarrote. It is a sort of culinary hybrid between pulque and mescal, but it is unclear which agave species was used. This information could be crucial to recover this apparently lost food, which could be a lost knowledge to humanity [The most striking result is the current absence of mention of a traditional fermented beverage made with mesquite pods reported by Bruman and a beverage named y Pineda , which whumanity ,190,191.pulque and mescal, (2) there is a systematic oversampling in the same localities for most of the studies; for instance, five pozol studies are performed in the same locality and this fact does not allow characterizing the potential area of this beverage, and (3) there has not been regular monitoring of the production of some of these beverages, and, consequently, it has not been attended before.A first sight of maps in We therefore propose that deeper studies are still necessary to identify, characterize, and monitor the actual and potential production areas of TMFB. This task will require fieldwork, information about the distribution of the substrates related to each beverage, and a careful characterization of localities where it is produced, using approaches from ethnobiology and ethnography that could bring insights about the possible processes of cultural erosion and the effects of market pressures and cultural discrimination over these marginal products. Such studies could also be the way to promote maintaining or the revival of these products ,192. TheIn recent years, the awareness to protect traditional relationships between humans and environments has promoted the development of systems of legal protection and biocultural management around the world . Numeroumescal production, for instance.Food systems are part of complex bodies of knowledge, most of them constructed empirically  and tranThe Mexican government signed the Nagoya Protocol in 2011 and was ratified in 2012. This is the major international regulation system about access to genetic resources and fair and equitable sharing of the benefits derived from their use, which emerged from the Convention on Biological Diversity. However, it was not until 2014 that the Nagoya Protocol was enacted as a supreme law in article 133 of the Mexican Constitution. The main aim of this protocol is favoring incentives for the conservation of biological diversity, the sustainable use of its components, and the prevention of misappropriation of genetic resources and traditional knowledge on them, which is relevant for a bioculturally mega-diverse country such as Mexico.Since Indigenous peoples hold traditional food system knowledge, interinstitutional and intersectoral initiatives would make more significant contribution to increase the potential of these resources, but local people should be protagonists. The documentation of traditional food systems is urgent since knowledge of food harvesting and preparation is rapidly disappearing. It is therefore crucial to enhance the public policies to support the maintenance of this traditional knowledge.Agave rhodacantha classified by the Mayo people as San Antoneña and El Chino were used in the fermentation on an agave beverage called yocogihua in southern Sonora and northern Sinaloa. Most commercial production of mescal in this region blinked out during the Ley Seca that began in Sonora in 1916, but remarkably the persistence of these varieties was documented off and on from 1888 to 1965 when the last plantation of them was closed. Nevertheless, the Mayo who worked in the plantation rescued or dispersed them to rancherias neighboring the plantation in Masiaca, Sonora, thus several fencerow plantings of A. rhodocantha persist nearby, even though it is otherwise rare except for one other location in Sonora. The recovery of these cultivars and possible reintegration into the contemporary production of culturally distinctive fermented beverages in Sonora is now being undertaken [bingarrote and bingui remain used for an analogous beverage today that is rarely produced in the state of Guanajuato where A. salmiana varieties are still in use for both pulque and mezcal production [bingarrote and bingui production near San Miguel de Allende could help to rescue both the plant genetic resources and the traditional cultural practices historically linked to them.The conservation of species and varieties involved is equally important. Cultural historians, ethnobiologists, and the general public should become aware of the distinctive varieties of plants used as substrates for fermentation, their conservation, and recovery. For instance, the varieties of dertaken ,209. As For local knowledge associated with microorganisms, the situation might be overly complex and difficult to attend, as certain microorganisms are not cultivable and not easy to identify. The lack of a microorganism’s accessions in collections, the low characterization of the selection mechanisms, and the implications of this selection over genetic and phenotypic traits in microorganisms can make the establishment of protective laws or conservation policies on this topic challenging. To understand the crucial roles of microorganisms on the TMFB process, we need to know their ecological niches, population dynamics, and relationships between microbiome and environments and microbiome and the selection process.Mexico has a unique gastronomic culture characterized by its multiethnic, high biocultural diversity and long cultural history. Biological diversity is used in traditional Mexican cuisine, and, undoubtedly, Mexican fermented beverages are intimately associated with plant products that have been managed and domesticated in Mexico, as it can be seen in the relevance of fermented beverages based on maize, agave sap and stems, or cacti fruits. However, this is also the case of dozens of wild species that are involved in traditional beverages. Foreign technologies, tools, and plants have been integrated into the Mexican foodscapes, and this fact has promoted a diversification of beverages and a diversification of sensory qualities acceptable to a wide range of palates. TMFB are clear examples that traditional knowledge is a dynamic process that is constantly changing, adjusting, and evolving.Agave group, leaving apart other beverages that are rarely studied, thus limiting the action to combat the endangered permanence of these products and the local food systems. The study of fermented beverages under transdisciplinary approaches is fundamental to provide information to construct regulatory frameworks or proposals to protect the diversity that TMFB involves in maintaining the complex interactions between humans, plants, and microorganisms. We expect that these examples demonstrate how biocultural conservation and restoration can be tangibly integrated into the protection and promotion of the Mesoamerican gastronomic patrimony recognized by the UNESCO and the routes of research needed to achieve it.Traditional Mexican cuisine is an outstanding reservoir of genetic resources such as plants, animals, fungi, and, generally less considered, microorganisms such as yeast and bacteria which could play a relevant role for future nourishment applications. The TFMB are reservoirs of cultural diversity, practices, and worldviews of communities. However, studies have focused mainly on the biotechnological applications and on beverages of the


Mikania micrantha, a noxious invasive weed, in this sense provides an excellent opportunity to test the explanation. The genetic and epigenetic variation of 21 invasive populations of M. micrantha in southern China have been examined by using transposon display (TD) and transposon methylation display (TMD) techniques to survey 12 TE superfamilies. Our results showed that M. micrantha populations maintained an almost equally high level of TE‐based genetic and epigenetic variation and they have been differentiated into subpopulations genetically and epigenetically. A similar positive spatial genetic and epigenetic structure pattern was observed within 300 m. Six and seven TE superfamilies presented significant genetic and epigenetic isolation by distance (IBD) pattern. In total, 59 genetic and 86 epigenetic adaptive TE loci were identified. Of them, 51 genetic and 44 epigenetic loci were found to correlate with 25 environmental variables . Twenty‐five transposon‐inserted genes were sequenced and homology‐based annotated, which are found to be involved in a variety of molecular and cellular functions. Our research consolidates the importance of TE‐associated genetic and epigenetic variation in the rapid adaptation and invasion of M. micrantha.Why invasive species can rapidly adapt to novel environments is a puzzling question known as the genetic paradox of invasive species. This paradox is explainable in terms of transposable elements (TEs) activity, which are theorized to be powerful mutational forces to create genetic variation.  Mikania micrantha populations maintained an almost equally high level of TE‐based genetic and epigenetic variation and they have not been differentiated into subpopulations genetically and epigenetically. 51 genetic and 44 epigenetic loci were found to correlate with 27 environmental variables . Twenty‐five transposon‐inserted genes were sequenced and homology‐based annotated, which are found to be involved in a variety of molecular and cellular functions. TD is capable to identify the integration site of transposons in gene tagging  temperature and precipitation changes have effects on growth and reproduction mode; (b) the weed shows strong endurance to heavy metals; and (c) it appears sensitive to light environment change caused by vegetation coverage  is listed as one of the top ten worst weeds  5′‐TACTCAGGACTCAT‐3′ (−) 3′‐GAGTCCTGAGTAGCAG‐5′. MspI/HpaII‐adaptor: (+) 5′‐GATCATGAGTCCTGCT‐3′ (−) 3′‐AGTACTCAGGACGAGC‐5′. EcoRI‐adaptor: (+) 5′‐CTCGTAGACTGCGTACC‐3′ (−) 3′‐CATCTGACGCATGGTTAA‐5′.We used a modified TMD protocol as previously described , 0.5 μl of each primer (10 μM), and one unit Taq DNA polymerase. The PCR conditions were as follows: initial denaturation at 94℃ for 30 s; followed by 23 cycles at 94℃ for 30 s, 56℃ for 30 s, and 72℃ for 1 min; and a final extension step at 72℃ for 10 min. Monomorphic bands in both HpaII and MspI digestions were regarded as nonmethylated CCGG sites, whereas presence–absence matrices were used to accommodate methylated types.Pre‐amplification PCR was conducted with one TE‐specific primer and another primer complimentary to the core sequence of the Similar to the above pre‐amplification, selective amplification was performed to acquire different chimeric sequences (TE/flanking DNA) by using various combinations of adaptor primers with different selective bases and a specific 5‐FAM end‐labeled TE primer. The selective PCR conditions were as follows: initial denaturation at 94℃ for 30 s; 15 cycles of 94℃ for 30 s, 66.5℃ for 30 s (decreasing 0.7℃ per cycle), and 72℃ for 1 min; followed by 23 cycles of 94℃ for 30 s, 56℃ for 30 s, and 72℃ for 1 min; and a final extension step at 72℃ for 10 min. The PCR products were electrophoresed in an ABI 3730 DNA analyzer (Applied Biosystems) with the GeneScan 1200 LIZ Internal Size Standard (Applied Biosystems).MseI, rather than HpaII and MspI used in TMD. The MseI‐adaptor pair and the TE‐specific primer were shown in Table Transposon display was achieved using the methylation‐insensitive restriction enzyme Transposon methylation display and TD bands were extracted and reamplified (using the same PCR conditions as in the selective amplification reaction) and sequenced. Respective restriction enzymes, adaptors, and primers of TMD and TD were listed in Table 2.4M. micrantha invasive populations were air‐dried and ground to pass through a sieve with an aperture size of 1 mm and 0.2 mm. Soil moisture was determined by oven‐drying for 6 hr at 105℃. The measurement of soil pH and electrical conductivity was performed in a solution of soil mixed with water at a ratio of 1:5 (W:V) using a pH meter  and a conductivity meter , respectively. Soil organic matter content was measured by using the potassium dichromate volumetry method. Soil total nitrogen was quantified by Kjeldahl method using KjeltecTM 8400 Analyzer Unit (Foss) after digestion at a ratio of 1:10 (W:V) soil to H2SO4. Total carbon was determined in a Total Organic Carbon Analyzer (Shimadzu) under the burning conditions of 720℃. Soil K, Ca, Na, Mg, Al, P, S, Si, Fe, Mn, Zn, Cu, Pb, Cr, As, Se, Ni, and Cd were assayed using an Inductively Coupled Plasma Optical Emission Spectrometer  after acid digestion. Experimental treatments without soil samples were served as control. All the measurements were repeated three times.Soil samples collected from 21 2.5GeneMarker v2.2 , Shannon information index (I), and unbiased expected heterozygosity (uHe), as well as percentage of permethylation (%Per), hemimethylation (%Hemi), and nonmethylation (%Non).GenAlEx v6.5  through Arlequin v3.0 with 1,000 random permutations  that significantly deviated from the expected ratio (1:1) at mutation–drift equilibrium (p < .05).Bottleneck v1.2.02  .To detect linkage disequilibrium (LD) between two loci, we implemented TASSEL v3.0  were assessed based on Structure Harvester (http://taylor0.biology.ucla.edu/structureHarvester/)  to further find the eigenvalues and eigenvectors of population correlation matrix in a multidimensional dataset , nine distance intervals were set: 0–10, 10–20, 20–30, 30–40, 40–50, 50–60, 60–70, 70–80, and 80–90 km. The upper and lower 95% confidence intervals and level of significance (P) were established for each observed value and each distance class using 9,999 Monte Carlo permutations of individuals among different distance classes. SPAGeDi v1.3d was used to further assess fine‐scale spatial genetic structure through estimating Sp statistic based on Sp = −kb/(1 − θ1)   between two pair populations, the significance of which was estimated through 9,999 permutations.To survey spatial genetic structure (SGS), we used Moran's http://www.rubic.rdg.ac.uk/~mab/stuff), a modification for dominant markers of the software developed by Beaumont and Nichols  was ≥2. As a widely used frequentist method based on a symmetrical island model, Dfdist enables to detect signatures of selection by comparing observed STF of each locus to coalescent‐simulated neutral global distributions by excluding 30% of the highest and lowest STF values. STF values above the upper 99.5% quantile were considered as outliers. The graphic was obtained using R v3.2.5  and Wald tests with a Bonferroni correction for multiple comparisons . Based on the above results, we further applied the univariate linear regression model in SAM v4.0 to estimate the correlation (r2) between outliers and environmental variables , ten ecological factors , and 24 soil factors  with a cut‐off E‐value of 10−10 and the genomic DNA databases of H. annuus r1.2 genomes in Phytozome v12.1 . Gene ontology (GO) was used to achieve annotations including molecular function, cellular component, and biological process ontologies. We further used Kyoto Encyclopedia of Genes and Genomes  and EuKaryotic Orthologous Groups  to perform pathways annotation and predict possible functions, respectively.For sequences flanking TEs, we searched against Nr .To guarantee the accuracy of results, we separately performed repeatability test using two independent TD and TMD capillary electrophoresis. The results showed that both TD and TMD had high repeatability (the independent sample I) of 0.27–0.37, and unbiased expected heterozygosity (uHe) of 0.18–0.25. At the species level, the total loci reached 1,802. The percentage of polymorphic loci, Shannon information index, and unbiased expected heterozygosity were 100%, 0.40, and 0.25, respectively (Table p < 0.001); and variation among populations within regions and among regions were 7.2%–17.24% and 3.42%–11.64% (p < 0.001), respectively  were 97.78%–100% and 0.21–0.29, respectively, while unbiased expected heterozygosity (uHe) was 0.13–0.19. At the species level, we acquired 2,676 epiloci with the percentage of polymorphic epiloci of 100%, Shannon information index of 0.29, and unbiased expected heterozygosity of 0.17. The percentage of permethylation (%Per), hemimethylation (%Hemi), and nonmethylation (%Non) were 31.91%–41.54%, 24.80%–30.14%, and 30.30%–38.47%, respectively. Among the 12 TE superfamilies, MITE‐, MUDR‐, and Tc1‐Mariner‐TMD exhibited low level of methylation (Table p < 0.001), and only 2.12%–9.77% was partitioned among regions (p < 0.001)  appeared in CACTA superfamily, while the lowest in MITE. In contrast, CACTA superfamily had the lowest hemimethylation (%Hemi), while LINE had the highest. Tc1‐Mariner and PIF‐Harbinger had the highest and lowest nonmethylation (%Non), respectively. Of note, Tc1‐Mariner exhibited the lowest genetic differentiation among populations and regions, while MUDR and PIF‐Harbinger showed the highest genetic differentiation among populations and regions, respectively. As for epigenetic differentiation among populations, Tc1‐Mariner was still the lowest, and LARD was the highest. LARD‐TMD maintained the highest epigenetic differentiation among regions, while Ty1‐Copia and Tc1‐Mariner had the lowest.For each TE superfamily, MUDR‐TD presented the highest Shannon information index and unbiased expected heterozygosity, while the lowest values occurred in SINE. For TMD, the highest percentage of permethylation (p < 0.05). In contrast, partial Mantel tests showed that six TEs had a significant pattern of IBD under the control of epigenetic variation, whereas seven TEs yielded a significant pattern of IBD under the control of genetic variation.Mantel test of TD and TMD revealed that isolation by distance occurred in six TE superfamilies: Ty1‐Copia, Ty3‐Gypsy, LARD, SINE, MITE, and CACTA (p < 0.05)  and the stepwise mutation model (S.M.M.), respectively (5) Table . For TMDM. micrantha populations by using TD and TMD. The optimal cluster K‐value was selected as two with Structure Harvester. Hong Kong and Shenzhen populations formed cluster I, whereas the other populations comprised cluster II  displaying linkage disequilibrium based on TD data (TE‐TD‐LD), of which PIF‐Harbinger accounted for the largest proportion. By contrast, twelve TE superfamilies all had epiloci (110) displaying linkage disequilibrium based on TMD data (TE‐TMD‐LD); of them, LARD had the highest proportion, while Ty1‐Copia and MITE had the lowest. Percentage of TE‐TD LD to total loci and to adaptive loci ranged from 0% to 1.30% and from 0% to 43.94%, respectively. Percentage of TE‐TMD‐LD to total and adaptive loci was from 0% to 0.67% and 0 to 100%, respectively , Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and EuKaryotic Orthologous Groups (KOG). Based on GO terms, they were assigned to 36 categories, including four cellular components, 15 molecular functions, and 17 biological processes Figure .The annotation results indicated that Ty1‐Copia and Ty3‐Gypsy separately inserted into genes related to probable S‐adenosyl‐L‐methionine‐dependent methyltransferase (SAM Mtase) superfamily protein and ubiquitin‐conjugating enzyme family protein, genes related to MITE insertion were probable (−)‐β‐pinene synthase and probable cryptochrome 1, and genes with insertion of Helitron, CACTA, and PIF‐Harbinger were associated with probable ribonucleotide reductase 1, auxin response factor, and kelch repeat superfamily protein, respectively Table .44.1M. micrantha, and the relative contribution of genetic and epigenetic variation to genetic differentiation has also been dissected. Generally, genomes of Asteraceae plants have been observed to have abundant transposons, which can cause genome size variation and impose influence on growth and development in multiple aspects  by performing AFLP and MSAP analysis. Following this line, it will be of considerable interest to characterize the association between TE epigenetic variation and phenotypic variation in the M. micrantha populations.Since it is hard to examine all TEs, representative candidate TEs were selected in this study, which focuses on high‐copy‐number families, low‐copy‐number LTR retrotransposons, and other TEs such as MITEs, LINEs, and SINEs. Their genetic and epigenetic variation have been detected by using the method of TD and TMD in s et al.  have sugAlternanthera philoxeroides); Liu et al. . And similar finding was noted for Japanese kotweed too. Nevertheless, nearly the same level of both types of TE variations was detected in M. micrantha, which deserves further study.Moreover, epigenetic variation is generally assumed to be greater than genetic variation in plants due to the frequent occurrence of retrotransposons  analysis was conducted. For BSGS, a difference was observed between the positive spatial correlation patterns established from TD or TMD data. Namely, a positive correlation was revealed at both class 0–10 and 10–20 km when using TD, whereas a positive correlation can only be found at class 0–10 km using TMD. For FSGS, by contrast, a similar pattern was uncovered based on TD or TMD data. A positive correlation was identified at levels of intra‐group, 0–0.15, and 0.15–0.30 km by using either method. A similar positive spatial (epi) genetic structure pattern was also observed within 300 m. Causal factors that contribute to the spatial (epi) genetic structure of Moreover, significant genetic and epigenetic IBD patterns were observed. This is in accordance with the findings by using MSAP  and introns and intergenic sequences. In total, we identified and annotated 25 genes with TE insertions  A gene probably coding SAM‐Mtase, a core enzyme in phenylpropanoid and flavonoid metabolism, is inserted with Ty1‐Copia. The gene may function in plant disease resistance and development ; Supervision . Qiqi Huang: Investigation . Zhen Wang: Data curation ; Writing‐review & editing . Ting Wang: Funding acquisition ; Supervision .Figure S1Click here for additional data file.Figure S2Click here for additional data file.Figure S3Click here for additional data file.Appendix S1‐S2Click here for additional data file.


H3K27M-mutant diffuse midline glioma is a rare childhood cancer originating in midline brain structures. The H3K27M mutation substitutes an amino acid on histone H3 that promotes gene expression and tumor growth. This cancer has a dismal prognosis and requires new and better treatment approaches. This review discusses controversies regarding tumor biopsy and summarizes molecular tumor characteristics that are therapeutic targets. We describe preclinical studies and clinical trials utilizing immunotherapy, radiation, and chemotherapy against this cancer. H3K27M-mutant diffuse midline gliomas (DMGs) are rare childhood central nervous system tumors that carry a dismal prognosis. Thus, innovative treatment approaches are greatly needed to improve clinical outcomes for these patients. Here, we discuss current trends in research of H3K27M-mutant diffuse midline glioma. This review highlights new developments of molecular pathophysiology for these tumors, as they relate to epigenetics and therapeutic targeting. We focus our discussion on combinatorial therapies addressing the inherent complexity of treating H3K27M-mutant diffuse midline gliomas and incorporating recent advances in immunotherapy, molecular biology, genetics, radiation, and stereotaxic surgical diagnostics. T,6. T5,6]H3K27M-mutant diffuse midline gliomas typically arise in the pons, infiltrate and expand throughout the brainstem, and cause progressive symptoms. Consequently, symptomatology of these tumors depends on the size and extent of infiltration. Nevertheless, patients typically present with hydrocephalus, pseudo-bulbar symptoms , lower cranial neuropathies, and headaches .Characteristic clinical symptoms and pathognomonic radiographic findings lead to a presumptive H3K27M-mutant diffuse midline glioma diagnosis. Conventional magnetic resonance imaging (MRI) typically demonstrates a hyperintense signal on T2-weighted images and expansion of the pons (taking up at least 2/3 of the pons) and sometimes adjacent brainstem. Enhancement patterns for H3K27-mutant diffuse midline gliomas vary and may not always be present at disease presentation A,C 11]..11].Histological and molecular tumor characteristics traditionally have not guided treatment strategies in H3K27M-mutant DMG . HoweverSince repeat biopsy is rare for DMG patients, recent advances have suggested that liquid biopsies (through serum) could measure treatment response by detecting H3K27M mutations in circulating tumor DNA ,21. SpecThe 2016 WHO classification of central nervous system tumors considers H3K27M-mutant diffuse midline glioma a high-grade lesion (WHO grade IV) ,27. The Our current understanding of the molecular biology of H3K27M-mutant DMG has resulted from the analysis of tumor tissue obtained through stereotactic surgical biopsy or postmortem tissue procurement . In a reH3F3A and HIST1H3B/C genes, respectively, with over 70% occurring in H3F3A . In. Inp = 0Kline et al. recently reported a study of the efficacy of combination re-irradiation and PD-1 inhibition using nivolumab in 31 patients with H3K27M-mutant diffuse midline glioma . PatientTechnological and neurosurgical advancements in stereotactic MRI-guided infusion catheter placement, catheter design, and drug distribution monitoring have paved the way for clinical studies and trials of intratumoral drug delivery via convection-enhanced delivery (CED) . CED is Pseudomonas exotoxin in five pediatric patients with H3K27M-mutant diffuse midline glioma -8H9) into the brainstem of pediatric patients with H3K27M-mutant DMG [124I]-8H9 administered by convection-enhanced delivery. No dose-limiting toxicities occurred with any dose, and the maximum-tolerated dose was not reached. Overall survival in this group was slightly better than historical controls (15.3 months). Survival increased more in patients in higher-dose cohorts. Two clinical trials using CED to treat H3K27M-mutant diffuse midline glioma have recently been reported. Heiss et al. investigated the safety, infusion distribution, and potential efficacy of CED of IL13–e glioma . They obtant DMG . The pri124I]-8H9 delivered by CED to the brainstem tumor with repeated CED infusions in pediatric patients [These studies confirm that CED can be used to deliver therapeutic agents safely and effectively to the brainstem in high intratumoral doses without systemic exposure or toxicity. Several other clinical trials are building upon the studies mentioned previously ,72,75. Apatients . Anotherpatients ,81,82. Tpatients . Tumor-localized therapies that do not require tumor catheters have also been proposed for H3K27M DMG. Super-selective intraarterial cerebral infusion is a novel tumor-localized therapeutic platform. This technique typically requires a primer such as mannitol to temporarily open the blood–brain barrier during arterial infusion. Recently, 10 treatment-refractory DMG patients received a super-selective intraarterial cerebral infusion of bevacizumab and cetuximab with minimal toxicity. Overall survival in this Phase 1 trial NCT01884740) was 17.3 months from diagnosis, and several patients had radiographic responses . Magneti740 was 1Immunotherapy of H3K27M-mutant diffuse midline glioma is being tested as an adjuvant to traditional chemoradiation. Immunotherapy was safe when administered intravenously and concomitantly with radiotherapy ,72,74. BA review of recent studies and trials investigating various immunotherapeutic methodologies for the treatment of H3K27M-mutant diffuse midline glioma is shown in Since H3K27M DMG treatment remains largely palliative at this time, the potential benefit of treatment should outweigh treatment-related side-effects affecting the patient’s quality of life (QOL). Treatment plans must be developed compassionately and empathically, bearing in mind the currently limited effectiveness of H3K27M-mutant diffuse midline glioma treatment. The median survival for H3K27M-mutant diffuse midline glioma is 9–12 months, and 2 and 5 year survival rates are 10% and 1%, respectively [H3K27M-mutated diffuse midline glioma is a devastating childhood central nervous system tumor with a dismal prognosis. Consequently, there is a desperate need for novel and effective therapeutic strategies to improve care for affected children. Recent advances in understanding the molecular biology of H3K27M-mutant diffuse midline glioma have driven research efforts. Ongoing laboratory studies and clinical trials aim to develop better chemotherapy, radiotherapy, immunotherapy, and intratumoral targeted treatments for H3K27M-mutated diffuse midline glioma. Combination therapies hold great promise for improving the present poor prognosis for H3K27M-mutant DMG patients. Basic and translational research findings provide insights into pathology, disease mechanisms, and novel therapeutic strategies for clinical trials. An improved understanding of the effects of tumor genetics, epigenetics, along with the immune microenvironments linked to patient outcomes, may lead to treatments better tailored to these factors. Therapeutic agents must reach the tumor to be effective. Thus, improved delivery techniques and distribution of therapeutics to the tumor are vital to enhancing the treatment of this presently incurable disease.H3K27M-mutated diffuse midline glioma affects young children and has a dismal prognosis. Over the past 10 years, the name of this tumor was changed from DIPG to H3K27M-mutated diffuse midline glioma after its mutation was identified and its prognosis was found to relate better to the tumor’s mutation than to its histological appearance. Identification of molecular pathways proceeding from the H3K27M-mutatation to tumorigenesis has uncovered potential therapeutic targets. In vitro and animal models for H3K27M-mutated diffuse midline glioma provide the opportunity to perform preclinical studies with novel therapies. Clinical trials have showed that therapeutic platforms, such as convection enhanced delivery, blood-brain barrier opening, and immunotherapy, were tolerated well by patients with H3K27M-mutated diffuse midline glioma. Although clinical trials have not yet increased life expectancy, they paved the way for future trials using novel agents that showed efficacy in preclinical models. Patients with H3K27M-mutated diffuse midline glioma direly need better treatment. Scientists and clinicians are learning more about the molecular pathways driving tumor growth and invasion and are working tirelessly to develop more effective therapies for this devastating condition that improve the lives of affected patients.


There is growing research interest in learning the genetic basis of response and adverse effects with psychotropic medications, including antipsychotic drugs. However, the clinical utility of information from genetic studies is compromised by their controversial results, primarily due to relatively small effect and sample sizes. Clinical, demographic, and environmental differences in patient cohorts further explain the lack of consistent results from these genetic studies. Furthermore, the availability of psychopharmacological expertise in interpreting clinically meaningful results from genetic assays has been a challenge, one that often results in suboptimal use of genetic testing in clinical practice. These limitations explain the difficulties in the translation of psychopharmacological research in pharmacogenetics and pharmacogenomics from bench to bedside to manage increasingly treatment-refractory psychiatric disorders, especially schizophrenia. Although these shortcomings question the utility of genetic testing in the general population, the commercially available genetic assays are being increasingly utilized to optimize the effectiveness of psychotropic medications in the treatment-refractory patient population, including schizophrenia. In this context, patients with treatment-refractory schizophrenia are among of the most vulnerable patients to be exposed to the debilitating adverse effects from often irrational and high-dose antipsychotic polypharmacy without clinically meaningful benefits. The primary objective of this comprehensive review is to analyze and interpret replicated findings from the genetic studies to identify specific genetic biomarkers that could be utilized to enhance antipsychotic efficacy and tolerability in the treatment-refractory schizophrenia population. Genetic testing is increasingly utilized to identify genetic biomarkers for optimizing the efficacy and tolerability of psychotropic drugs, especially antidepressants. However, genetic testing is also being requested to enhance the effectiveness of antipsychotic drugs, which is especially true for the treatment-refractory schizophrenia population, who frequently experience irrational polypharmacy at high dosages with significant adverse effects, generally without much therapeutic benefit. Currently, the most evidence in supporting genetic assays is based on pharmacogenetic (PG) studies, which utilize a candidate gene(s) approach investigating the effect(s) of genetic polymorphism(s) on efficacy, tolerability, and/or safety of antipsychotic drugs . Most clThe data from PG studies are clinically utilized at the individual level to predict and optimize the response to antipsychotic drugs while preventing or minimizing adverse events. A drug’s response or tolerability can be affected by genetic polymorphisms in PK factors, which determine the concentration of a drug at its site(s) of action, and PD factors, which determine a drug’s response or tolerability at its molecular targets. However, these distinctions are rather arbitrary, as changes in a drug’s concentration at the site of action  are always associated with changes in a drug’s efficacy and/or tolerability  at its site(s) of action. The following section will review the PK and PD genetic findings from the pharmacogenetic studies, followed by a brief discussion of pharmacogenomic studies, commercially available assays, and future directions.Genetic variance in drug-metabolizing enzymes, such as CYP enzymes, represents most of the PK biomarkers. The genetic polymorphisms of CYP enzymes have produced one of the most replicated and clinically relevant findings in patients who develop adverse effects on routinely administered dosages of an antipsychotic drug. A similar statement cannot be made for antipsychotic efficacy, probably because there is no apparent relationship between plasma levels of an antipsychotic drug and antipsychotic response with the exception of clozapine. In this context, CYP2D6 is one of the most clinically relevant enzymes; despite making only 2% of all CYP enzymes in the liver, CYP2D6 is involved in the metabolism of about 25% of several commonly used psychotropic agents, including antipsychotic drugs . About 61A2 has also been associated with adverse effects due to an increase in plasma levels of antipsychotic drugs that are substrates for this enzyme, such as clozapine and olanzapine [Deficient activity of CYP enzyme anzapine ,42,43. Ianzapine . One stuanzapine , while oanzapine ,22. Howeanzapine ) has beeanzapine  and clozanzapine .D2−141C Del and TaqI A2 allelic variants have been associated with the inadequate antipsychotic response across various ethnic groups [D2−141C Del and TaqI A2 allelic variants and antipsychotic response [Antipsychotic efficacy across different antipsychotic drugs has been strongly linked with genetic variance in dopamine-2 receptors (DRD2). More specifically, c groups ,50,51,52response   102-C/C genotype, which was associated with reduced antipsychotic efficacy in Caucasian patients [-HT2A −1438-A/A genotype [5-HT2A −1438-G allele was associated with psychotic relapse [5-HT1A −1019G), has been associated with lower antipsychotic efficacy in various ethnic groups [5-HTT LPR (serotonin transporter-linked promoter region) and improvement in negative symptoms of schizophrenia was observed in one study [Specific polymorphisms in serotonin receptors have also been linked with the antipsychotic response, especially serotonin-patients ,62  variant  has also been associated with lower risperidone efficacy in two studies [BDNF) in two studies [DRD4 variance [Risperidone is another second-generation antipsychotic drug, which has shown decreased antipsychotic efficacy in patients with DRD2 Ser311  variant  studies ,116 and  studies  Table 1COMT vari studies , Korean  studies , and Jap studies  patients studies ,121. Howvariance . Other gvariance ,128,129.D3Ser9Gly [5-HTT LPR [Olanzapine is another commonly used second-generation antipsychotic drug with reports of an association between its efficacy and DRD3 variant 3Ser9Gly ,131, whi3Ser9Gly ,131. How3Ser9Gly , suggest3Ser9Gly . In term-HTT LPR  and HTR6-HTT LPR  associat-HTT LPR , highlig-HTT LPR  associat-HTT LPR ,138.D2 TaqI variants and the efficacy of aripiprazole in Korean and Chinese patients [Although aripiprazole is classified as one of the newer second-generation antipsychotic drugs, it is the first antipsychotic drug with partial agonist activity at D2 receptors and 5HT1A receptors . A couplpatients ,141. In The genetic data for antipsychotic tolerability is not as consistent as those for antipsychotic efficacy, except for weight gain. The margin for controversial results is much higher than those from the efficacy studies, as documented below.DRD3 and CYP 17A1 genotypes and EPS [Although genetic polymorphisms in CYP enzymes are grouped under PK biomarkers, it is worth mentioning here that any change in a drug’s metabolism will eventually be expressed pharmacodynamically. Thus, the poor metabolizers for CYP2D6 have a higher risk for developing EPS due to increased plasma levels of antipsychotic drugs that are CYP2D6 substrates ,19,20,21 and EPS . Another and EPS . A coupl and EPS ,171. How and EPS ,173,174. and EPS ,147,175  and EPS ,174. The and EPS ,177 as w and EPS  patientsGenetic variance in the serotonergic system has also produced inconsistent results; some reports have documented associations between HRT2A polymorphisms and TD ,179,180,Although there is not much research investigating the role of genetic variance on antipsychotic-induced hyperprolactinemia, any DRD2 polymorphism that increases the risk for EPS will also increase the risk for hyperprolactinemia, as both adverse effects are mediated by D2R blockade. In this context, one study did report 40% higher prolactin levels in patients with DRD2*A1 allele than those without . Interes5-HTT LPR) was significantly associated with weight gain and obesity in the Caucasian population [Serotonin is one of the main neurotransmitter systems that control feeding behavior in the hypothalamus and is targeted by the second-generation antipsychotic drugs via their blocking effects on 5HT2C receptors . Althougpulation ,221, butpulation .Although earlier studies did find an association between weight gain and genetic variance in DRD2 ,222 or DFew studies have reported a significant correlation between genetic polymorphism in melanocortin 4 receptors (MC4R) and antipsychotic-induced weight gain ,226, whiCYP2D6 [25 [One study reported a correlation between antipsychotic-induced weight gain and polymorphism in insulin-induced gene 2 , but a cCYP2D6 ,245, mulCYP2D6 , methyleCYP2D6 ,247, perCYP2D6 , synaptoP2D6 [25 , and tumP2D6 [25 ,251.NQO2) polymorphism [CYPD26 [Agranulocytosis is a rare but severe and potentially lethal adverse effect associated with clozapine use. Pharmacogenetic studies have reported strong associations between polymorphisms in the major histocompatibility complex and clozapine-induced agranulocytosis ,253,254.morphism  and negamorphism ,258. An morphism  but not morphism  or with ANKS1B) and contactin-associated protein-like 5 were linked to the efficacy of olanzapine and risperidone in terms of negative symptoms [MC4R) gene associated with weight gain in patients undergoing the first trial with antipsychotic drugs, which is consistent with a region previously identified by large-scale GWAS of obesity in the general population [MC4R in extreme SGA-induced weight gain and related metabolic disturbances. Most recently, polygenic risk scores derived from significantly associated SNPs with schizophrenia patients of European ancestry were found to inversely correlate with the antipsychotic response [These studies have primarily explored the effects of genetically mediated PD differences in a drug’s response and/or adverse effects through a systematic assessment of genes, their products, and individual variation in gene expression and function. In this context, GWAS are most useful to generate hypotheses that can be later confirmed in future studies. These studies explore associations between multiple genes and psychotropic drugs ; therefosymptoms . In addisymptoms , GABAergsymptoms , and HLAsymptoms . Anotherpulation . These dresponse . It willresponse .SLC26A9), DRD2, G protein-coupled receptor 137B, carbohydrate sulfotransferase 8, and interleukin1-alpha (IL1A) was associated with improvements in various neurocognitive domain areas. A significant result was also found for the variant rs286913 in the ETS homologous factor related to the effects of ziprasidone on vigilance. Furthermore, the presence of rs11240594 in the SLC26A9 gene and rs11677416 in the IL1A gene was linked to olanzapine-induced improvement in processing speed and working memory.Antipsychotic treatment in a subset of 738 schizophrenia patients from the CATIE study  polymorpAmpliChip™ is the only FDA-approved genetic test, which is a microarray-based product to assess the activity of CYP2D6 and CYP2C19 and can be helpful in a large number of psychiatric patients as multiple psychotropic drugs are metabolized by these two CYP enzymes. Genetic testing for CYP2D6 is among the most clinically relevant investigation, as several important psychotropic drugs, including antipsychotic drugs, such as haloperidol, perphenazine, and risperidone, are metabolized by this enzyme. Following are the major resources and genetic assay companies that offer genetic testing for psychotropic drugs.These assays offer genetic testing for multiple genetic biomarkers  for treatment response and/or tolerability identified in other studies to facilitate the selection of effective psychotropic medications. The clinical utility of these combinatorial genetic assays has primarily been tested in underpowered and open-label studies. Thus, these assays with specific panels for different groups of psychotropic drugs are based on findings from other studies using a candidate gene(s) approach. Although there is no specific genetic assay for antipsychotic drugs, combinatorial genotyping of genetic biomarkers is used to optimize the efficacy and tolerability of antipsychotic drugs, especially in the treatment-refractory population. In this context, genetic variance in PK biomarkers (primarily the CYP enzyme system) has been clinically helpful to optimize antipsychotic treatment. Most genetically relevant CYP enzyme assays for antipsychotic drugs include CYP1A2, CYP2D6, and CYP2C19. ®  combinatorial assays provide coverage for about 50 PK alleles, including those for CYP2D6, CYP2C19, CYP2C9, CYP2B6, CYP3A4, and CYP1A2, and some PD genes . On the basis of information on these genetic biomarkers, an individualized report is created which divides psychotropic medications into a green bin for recommended use, a yellow bin for use with caution, and a red bin use with extreme caution and frequent monitoring.The GeneSightTM assay (Genomind®) also provides testing for PK biomarkers  and PD markers, . Like the GeneSight report, each patient’s results are provided to the ordering clinician, along with suggested therapeutic options.GeneceptDrug-Metabolizing Enzymes and Transporters (DMET™) Plus Solution is one of the largest commercially available genetic assays for about 2000 PK variants across multiple genes. The DMET™Plus Solution was developed as a platform to identify genetic variance and has not been tested for its efficacy in enhancing clinical outcomes with psychotropic drugs.One of the most important goals for future genetic research in psychopharmacology will be to replicate and validate results from small sample genetic studies to resolve inconsistent results. However, these goals can only be accomplished by large, prospective, well-conducted multisite clinical trials such as genome-wide association studies to allow subgroup analyses and provide control for demographic, clinical, and environmental factors while close monitoring for medication adherence. In this context, the clinical trial network model used in oncology and cardiology has already been initiated in psychiatry, including Implementing Genomics in Practice (IGNITE), the Dutch Pharmacogenetics Working Group, and the Pharmacogenomic Resource for Enhanced Decisions in Care and Treatment. These large-scale initiatives will offer an effective tool to explore the relationship between efficacy and tolerability of various antipsychotic drugs and multiple genetic variants to generate hypotheses that could be tested in hypothesis-driven randomized controlled trials to improve and expand on currently available genetic biomarkers. A minimum set of genes and alleles is required for consistent pharmacogenomic decision-making before combinatorial genetic assays can be used beyond the treatment-refractory population. With the growing research in this area, it is only a matter of time that these efforts will help develop comprehensive combinatorial commercially available assays and toolkits with clinical utility beyond the treatment-refractory patient population facilitating the application of bench research to the clinical bedside. Equally importantly, clinicians must be trained and updated to enhance clinical application and interpretation of results from the existing and ongoing genetic studies with psychotropic medications, including antipsychotic drugs.


Prunus armeniaca L.) is an important temperate fruit crop worldwide. The availability of wild apricot germplasm and its characterization through genomic studies can guide us towards its conservation, increasing productivity and nutritional composition. Therefore, in this study, we carried out the genomic characterization of 50 phenotypically variable accessions by using SSR markers in the erstwhile States of Jammu and Kashmir to reveal genetic variability among accessions and their genetic associations. The genetic parameter results revealed that the number of alleles per locus (Na) ranged from 1 to 6 with a mean Na value of 3.89 and the mean effective number of alleles (Ne) per locus 1.882 with a range of 1.22 to 2. Similarly, the polymorphic information content (PIC) values ranged from 0.464 to 0.104. The observed heterozygosity (Ho) (0.547) was found to have higher than expected heterozygosity (He) (0.453) with average heterozygosity of 0.4483. The dendrogram clustered genotypes into three main clades based on their pedigree. The population structure revealed IV sub-populations with all admixtures except the III sub-population, which was mainly formed of exotic cultivars. The average expected heterozygosity (He) and population differentiation within four sub-populations was 1.78 and 0.04, respectively, and explained 95.0% of the total genetic variance in the population. The results revealed that the SSR marker studies could easily decrypt the genetic variability present within the germplasm, which may form the base for the establishment of good gene banks by reducing redundancy of germplasm, selection of parents for any breeding program.Apricot ( Prunus armeniaca L) is one of the influential fruits of the Rosaceae family that is mostly produced in temperate climates. Apricots have been divided into seven eco-geographical groups based on their origin [Prunus genotypes [Prunus species and stone fruits [Prunus armeniaca L. In this part of India, no such type of study has been carried out so far to evaluate the genetic diversity of the Prunus armeniaca L from the whole temperate areas of Jammu and Kashmir through SSR markers. Therefore, this study was undertaken to examine the genetic diversity and population structure between 50 apricot genotypes  and to evaluate the degree of variation among and within eco-geographical groups and subgroups of apricot germplasm taken into consideration.Apricot  ranged from 1 to 6, with an average value of 3.89. Among the 46 markers, the highest number of alleles, 6 per locus, was realized with 14 markers, and the highest number of effective alleles (Ne) was observed, 2 per locus with five markers such as RPPG5-030, RPPG6-033, PacA10, PacA22 and PacA26. The PIC value varied in range from 0.104 to 0.464, with an average value of 0.320. Furthermore, the number of effective alleles (Ne) ranged from 1.1563 to 2 with an average value of 1.8821. The average observed homozygosity (Ho) was 0.4774, and varied from 0 to 1, whereas the average expected homozygosity was 0.5470 which ranged from 0.4947 to 0.8634. Similarly, the observed heterozygosity ranged from 0 to 1 with an average value of 0.5226, and expected heterozygosity (He) ranged from 0.1366 to 0.5053 and produced an average value of 0.4530. The overall average heterozygosity was 0.4483 and ranged from 0.1352 to 0.5. The Shannon’s diversity index (I) ranged from 0.2611 to 0.6931 with an average value of 0.6371, and the genetic differentiation (Fst) ranged from 0 to 0.08 with an average value of 0.0228. Different parameters showed a lot of variabilities indicating high genetic diversity.The Jaccard’s similarity coefficients between the germplasms were calculated for UPGMA clustering. The diverse group of 50 germplasms were divided into three primary groups based on their genetic similarity at a distance of 0.614 as cluster I, cluster II and cluster III .Cluster I consisted of 10 exotic genotypes divided further into two sub-clusters IA which included 8 genotypes  and IB, which contained two genotypes (G2 and G10). Cluster II was the largest and contained 39 genotypes that are mainly indigenous to Jammu and Kashmir and form two sub-clusters: cluster IIA and cluster IIB. Cluster IIA is further divided into two sub-clusters and contained 8 genotypes in the first cluster  and 26 genotypes . Cluster IIB contained 5 genotypes  and cluster III contained single genotype G28. The Jaccard’s similarity among the genotypes ranged from 0.508 to 0.867. The highest similarity 0.867 was observed between indigenous accessions G45 and G49, which are accessions from Banihal and Doda regions of Jammu province of J&K and the lowest similarity 0.508 was observed between exotic cultivar Hartley and indigenous accession G31 from Malapora area of Baramullah. A 3D clustering plot revealed that 50 accessions produce three clusters C-I and C-IIA and C-IIB. In addition, the results from 2D PCoA clustering  were conThe clustering pattern of apricot accessions from 2D and 3D PCoA plots and the UPGMA clustering graph were highly consistent. The UPGMA clustering tree graph provides abundant information and categorizes the accessions into different groups. The information produced by the 2D PCoA plot, although not sufficient, produced a flat and direct view of the relationship between different accessions as compared to 3D PCoA which provides sufficient information in different layers and directions. The combined result analysis of population structure through genetic similarity and PCoA provides valuable information to understand the genetic structure of the accessions.An investigation carried out for population structure utilizing marker data assisted in recognizing four (K = 4) genetically different sub-populations in 50 diverse apricot genotypes. Initially, we were unable to estimate the number of subpopulations as the LnP (K) values decreased from 1 to K = 2 and then increased at K = 3 and then again decreased at K = 4 before subsequently increasing at K = 5 to K = 8 and started again declined at K = 9, and finally at K = 10 increased a. Thus, Hence, in our population of 50 apricot accessions, we found 4 subpopulations. Subpopulations 1, 2, 4 contained 36 genotypes that were all indigenous, while as in the 3rd subpopulation, only one indigenous genotype was spotted, the remaining 13 of the genotypes found were exotic. This arrangement pattern was also revealed in the structure graph , depictiThe purpose of the analysis of molecular variance was to see if there was any genetic variation across populations as well as within populations. According to our results, 95% of the variance was observed within the population, whereas only 5% of the overall genetic diversity was identified between populations .Prunus genotypes [Microsatellite markers have been successfully employed by several studies to identify molecular genetic variation in apricot genotype collections and populations ,49,50,51enotypes . The numenotypes  in diffeenotypes . The aveenotypes  and 0.59enotypes . Bourguienotypes  observedenotypes  investigenotypes  also obsenotypes  observedenotypes  revealedenotypes , Raji etenotypes  and Zhebenotypes , Maghulyenotypes , Liu et enotypes , Gurcan enotypes  and Akpienotypes . The Micenotypes  found a enotypes , Tian-Mienotypes , Romero enotypes , Maghulyenotypes  and BatnAll genotypes were divided into three main clusters, cluster I, cluster II, and cluster III, with varying degrees of sub-clustering based on the dendrogram. Cluster I comprised ten accessions, the majority of which were exotic genotypes. Cluster I was subdivided into two sub-clusters, IA and IB, which contained eight and two genotypes. Cluster II contained 40 genotypes that are mainly indigenous to Jammu and Kashmir. The grouping of genotypes revealed by the principal coordinate analysis (PCoA), biplot and cluster dendrogram is similar and shows consistency of the results of the grouping of genotypes based on the geographic areas of the sample collection. The first two coordinates of PCoA contributed 68.43% of total genetic variability and the maximum share of this genetic variation is contributed by cluster first (C-I) and cluster IIB (C-IIB). Previously, apricot accessions were arranged using molecular markers according to their geographic origins ,22,24. RSTRUCTURE analysis of the population is a convincing approach to examine genetic relationships and ancestry of individuals within gene banks . The STRAMOVA revealed a 95% variation within populations and 5.0% of the total molecular variability between populations. Gomez et al.  and VendFresh young tender leaves during preflowering season from each accession were taken in a plastic bag from the field and flash frozen in liquid nitrogen to keep them at −80 °C until DNA extraction. The geo-referenced data, name and exact location of apricot leaf sample collection from the field are shown in The procedure described by Doyle and Doyle  was used2, 10 mM dNTPs, 1U Taq DNA polymerase, and both primer pairs. A thermal cycler  was used for amplification. The PCR amplification steps were executed as initial denaturation for 5 min at 94 °C followed by 35 cycles of 60 s at 94 °C denaturation, 49 to 58 °C for 60 s for optimal annealing temperature for different primers, 90 s at 72 °C for extension and final extension for 10 min at 72 °C followed by cooling at 4 °C. The procedure was performed three consecutive times with the same primers and genotypes to check out the reproducibility. The PCR amplification products and the 100 bp DNA marker were separated on 3% agarose gel with 0.5× TBE buffers using Ethidium bromide (EtBr) as a staining agent on the gel. The banding pattern of the amplified bands was examined under a gel documentation imaging system.PCR reaction was carried out in a 20 μL reaction mixture with 50 ng/µL DNA templates, 10X PCR buffer, 2.5 mM MgClFor all accessions, the composition of alleles and each microsatellite locus were used to calculate the total number of alleles. Indices of molecular characterization were statistically evaluated, including the expected heterozygosity (He), the observed heterozygosity (Ho), the effective number of alleles (Ne), Shannon’s information index (I), the coefficient of gene differentiation (Fst) by applying the POPGENE 1.32 ,73,74. IIn conclusion, our investigation has dispensed a broader context on genetic variability and core structure among apricot accessions in Jammu and Kashmir. The results revealed that the SSR marker studies could easily decrypt the genetic variability present within the germplasm. This was the first kind of study carried out in this area to distinguish exotic genotypes from indigenous genotypes via molecular markers and showed a high level of polymorphism. Genetic variability between exotic and indigenous genotypes can provide an excellent opportunity for new cultivar development through hybridization and advanced genetic tools such as molecular markers. These diversity analysis tools could be utilized for the establishment and collection of gene banks and core collections by reducing redundancy of germplasm, selection of parents for any breeding program and genome-wide association studies for mapping of different traits.