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Academic production about the main Chilean cities exceeds the existing documentation on intermediate cities, though they have shown interesting trends patterns in recent years which have changed the urban system in Chile. This paper aimed to analyze the urban growth processes in Chilean intermediate cities using Temuco as a case study. It begins with an historical look at the city and then mentions that in the last decades this kind of cities have undergone new forms of segregation associated to real estate activities such as private communities, rural residential properties and new localization of services which have resulted in a fragmentation of urban space, a phenomenon reinforced by the consolidation of some satellite cities. At the same time, many urban problems associated to poverty, like a standstill of the regional economy and environmental pollution have appeared, jeopardizing the sustainability of these spaces, questioning current development parameters. Most of urban literature shows that research has been focused on big metropolitan areas. Transformations resulting from globalized neoliberal policies have been studied preferentially in big capitals in Latin America: Santiago, Buenos Aires, Sao Paulo, Mexico city, Lima and Bogota. Geographica Helvetica, 66 2 , Wohnghettos der Privilegierten. Die Fragmentierung der brasilianischen Stadt. Matices: Zeitschrift zu Lateinamerika. Spanien und Portugal, 8 29 , Nuevas formas y viejos valores: urbanizaciones cerradas de lujo en Guadalajara. Cabrales Ed. Guadalajara: Universidad de Guadalajara. Revista Latinoamericana de Estudios Urbano Regionales, 28 85 , Although the study of intermediate cities has gained importance in the last decades Mertins, Mertins, G. Espacio y Desarrollo 12, Espacio y Desarrollo, 12, Intermediate cities in Latin America risk and opportunities of coherent urban development. Cities London, England , 21 5 , Huancayo metropolitano. Cities London, England , 31, Bellet, E. Melazzo, M. Llop Eds. Lleida: Universidad de Lleida. The studies about socio-spatial segregation are examples of the previous statement. El modelo de ciudad latinoamericana. Buenos Aires a la deriva: transformaciones urbanas recientes p. Buenos Aires: Editorial Biblos. Even though there are many common aspects between intermediate and big cities, some authors consider revising the effects of the variation in the urban scale of the socio-spatial phenomena because this can lead to think over the nature of urban development as it is understood up to these days Capel, Capel, H. Due to these facts, the aim of this research is to explore the urban development trends in Chilean intermediate cities during the last ten years, considering topics such as segregation, fragmentation and sustainability. The case study is Temuco, a city situated in the center of the southern zone in Chile, with many and accelerated transformations in the last decades. The methodological approach is a triangulation of primary and secondary sources: academic articles; data of national state institutions i. The information was analyzed using the triangulation of the sources mentioned above. The article is divided in five sections. Firstly, the changes taking place in the last decades in intermediate cities are analyzed focusing on the Chilean case and its development tendencies in depth. Secondly, the case study is presented together with a brief review of Temuco urban history. Thirdly, two of the most important trends that have characterized city growth in the last decades: segregation and fragmentation are reviewed. In the fourth place, an analysis of the economic, social and environmental problems that put at risk the urban sustainability of Temuco is made. In the fifth place, we present a review of Temuco case study, an open outline of the growing trends in intermediate Chilean cities in the last decades. World urbanization prospects: the revision. New York: ONU. This scenario differs from the one just some decades ago: in urban population was Desenvolupament sostenible. Tarragona: URV. The observed strong growth of urban population in the last decades not only sets up a strain on environment but it has also questioned the traditional definitions of a city. On the other hand, the physical transformations and of daily life in the cities have boosted the search of a definition out of the strict demographic and physical parameters which are usually used to define urban space, encouraging for term concepts closer to cultural and ontological traits Hiernaux, Hiernaux, D. If it is difficult to define a city it is even more complex to state a typology according to its size. The facts and city scales differ in every country. An intermediate city for Spain can be a small one for China or Brazil. Santiago de Chile: Ministerio de Vivienda y Urbanismo. According to the classification of the Ministry of Housing and Urban Planning, Chilean research focused on minor and largest intermediate cities are uncommon. Although a bigger interest for this kind of urban spaces is recognized, the prevailing dynamics seem unclear, it is necessary a general pattern which summarizes the Chilean trends, specially one accounting for the transformations originated by the neo liberal economy market. Notwithstanding, beyond the limitations due to the shortage of studies, some of them have made evident some trends that are taking place in urban growth in intermediate cities in Chile. Intermediate cities are strongly associated to regional and surrounding dynamics Concha et al. In Congreso urbano no-metropolitano p. When the economic conditions in the region are good, these cities tend to channel and gather population growth and new productive investments, becoming jointed axis for regional development. An example is Puerto Montt city with a strong population and housing growth due to the aquaculture activity boom in the region Rovira, Rovira, A. Puerto Montt: el paso de ciudad menor a centro de desarrollo interegional de una ciudad al sur de Chile. In this way, the relation with the surrounding is complementary and not of invasive-domination lacking of control as it is in large metropolitan areas. On the other hand, the insertion of real state capitals in intermediate cities seems to be smaller than in metropolitan areas. This is observed in the lack of gentrification processes in intermediate cities as compared with central or peri-central areas as the one observed in metropolitan areas Contreras, Contreras, Y. Real estate investments in intermediate cities are made in vertically built condominiums and they are located on structuring roads and consolidated places in a city, where have traditionally lived the upper socioeconomic groups Borsdorf, Borsdorf, A. El desarrollo urbano de Valdivia: estudio de caso en una ciudad mediana. There are some other reasons which have influenced intermediate cities growth. Burden, stress and insecurity of modern life in the city have motivated the search for quieter places to live, closer to nature but also near to urban life facilities. Patrones de crecimiento urbano en la Patagonia chilena: el caso de la ciudad de Coyhaique. Even though this type of sub-urbanization process it is also patent in Chilean metropolitan areas Borsdorf et al. Cities London, England , 24 5 , Revista , 24, Finally, as the penetration of real estate capital is lesser in intermediate cities, most of all in those in an economic vulnerable regional position, an important aspect of urban growth can be explicated with social housing building; the localization of them is peripheral because its construction is subordinated to market forces and the lower price of the land Borsdorf, Borsdorf, A. Meanwhile in intermediate cities subjected to strong population growth resulting of good economic regional conditions, as a general fact social housing building has been expelled out of the city boundaries giving rise to the development of satellite dormitory towns concentrating vulnerable population living with poor facilities and badly equipped Borsdorf et al. Barcelona: Scripta nova. Synthetizing, intermediate cities have followed some development patterns of big cities, however they seem to have some characteristics resulting not only of the relationship with their regional surrounding but also of the level of penetration of the economic capital in them. Historia de Temuco. Temuco: Ediciones Universidad de La Frontera. Aylwin Ed. Temuco: Universidad de La Frontera. Mapuche are indigenous people who had remained independent during Spanish conquest and the formation of Chilean nation. Afterward the first turbulent years, Temuco was considered as an important area as much as for the development of communications and for the exchange of assets and people between the central and southern region of the country. Accordingly, the state initiated the construction of new roads and railways enabling migration and trade between creole, pioneers and indigenous people Municipalidad de Temuco, Municipalidad de Temuco. Plan regulador comunal de Temuco: memoria explicativa. Temuco: Municipalidad de Temuco. The new inhabitants of Temuco and surroundings devoted themselves to forestry, agriculture mainly cereal and cattle, these activities were the main income of the city allowing a consolidation of an emerging export model. Later on, they added their own business creating a small commercial zone near the railway station, where they sold products and regional handicraft. The consolidation of the city and the initial economic boom provoked a fast population growth. Indeed, in Temuco had only 3, inhabitant most of them soldiers meanwhile in it was a prosperous urban center with 16, locals Pino, Pino, E. National and international migrations and new state institutions settled in the zone as a result of the successful military operation initiated some years before. The quick growth was a reason to define the first urban boundary around , a rule that may be considered the first tool for the city planning Navarrete, Navarrete, G. The fragmented city: changing patterns in Latin American Cities. The Urban Reinventors Online Journal, 2 9 , Los inicios de la insustentabilidad: problemas urbanos e institucionalidad en la ciudad de Temuco, Cuadernos de vivienda y urbanismo, 8 16 , There lived rural vulnerable population or mapuche descendants who had to face hard living conditions, like overcrowding, lack of the minimum standards of basic housing services drinking water, sewage or electricity and health problems plagues and diseases. On the other hand, the consolidation of the center of the city and the urban expansion towards the west, especially along Alemania Avenue, concentrated the construction of middle and upper class neighborhoods Figures 2 and 3. The liberalization of the urban land market in brought about a significant growth and increased real estate companies. Revista INVI, 20 54 , The reforms of urban policies have contributed to a transformation of the city. In the next subheading we are going to review two key trends to understand the development of the intermediate city in a neoliberal frame of segregation and segmentation. The installation of neoliberal policies in the city has produced and consolidated some dynamics that have strongly affected the configuration of the city, and also triggering new ones. These new urban reforms were for Temuco an impulse which helped to consolidate its urban development towards the west part of the city Municipalidad de Temuco, Municipalidad de Temuco. At the late eighties and early nineties, Alemania Avenue started not only a densification process, but also urban functions changed, mostly from a residential use to a mixed use residential — commercial. Nowadays, on Alemania Avenue there are several banks, restaurants, pharmacies, two shopping malls and even a Casino and a five stars hotel. Since the beginning of the 90s not only social housing were built in the city, as well new upper and middle class neighborhoods emerged in the peri-urban area occupying and changing the traditional land usage. In the case of Temuco companies such as Fourcade, Socovesa and Aconcagua are responsible for the construction of almost all new residential projects in the peri-urban area. These projects are mostly gated communities and pleasure lots. On the other hand, pleasure lots are single lots of no less than m 2 which are governed by the Law of , created to avoid the atomization of lands lots for farming. The speed of the recent city expansion can be interpreted looking at the building permits given by the municipality. Figure 4 shows the evolution of the approved surface of building permits for residential uses between and They have experienced an irregular pattern, which reached its peak in , but had no continuity decreasing over Santiago: Ministerio de Vivienda y Urbanismo Alameda. At first glance this fluctuating rhythm could be explained by the growth experienced in the surrounding localities, which belongs to the conurbation such as Labranza, Cajon or Padre Las Casas. Other hypotheses trying to explain this situation pointed out the slowdown of the economy and the weak situation both of labor market and the bad performance of some regional indicators such as the generation of investment, which are among the lowest indexes compared to the national average of major cities in the country. Also the lack of a properly connectivity plan could be one factor explaining this scenario PatagonLand, PatagonLand. Puerto Varas: Patagonland. All this projects are conceived and promoted as an alternative for people who want to live in a rural environment but at the same time enjoying the proximity to the city and its facilities. This kind of fragmented development is increasing and consolidating socioeconomic segregation and inequality, phenomena that could be observed in different degrees in most of the intermediate cities and metropolitan areas of Chile Sabatini et al. Revista Latinoamericana de Estudios Urbano Regionales, 29 87 , These segregations patterns observed in Chile do not differ from the rest in Latin America, this is associated to the phenomena of real estate operations performed by the companies in peripheral zones in the historic city center Sabatini, Sabatini, F. The segregation consequences for poor people are assorted, from social disintegration caused by exclusion feelings to the formation of subcultures deviated from the current norms of a society Sabatini et al. Documento de Trabajo, 37, p. Seducidos y abandonados: el aislamiento social de los pobres urbanos. The specific configuration of the type of segregation in an urban space, depends on of the particular traits of the society in which it is integrated, but as a general rule, socio-spatial segregation is aligned to socioeconomically factors. According to this, the phenomenon represents one of the most important dimensions helping to comprehend the occupation of every area in a city, considering the land value and also the accessibility to a given social sector to a space Romero et al. In: C. Hidalgo Eds. We can point out that as in metropolitan areas, in intermediate cities like Temuco gated communities and private neighborhoods have been built in the suburbs Figure 5 , however, these ones have been located far from the zones occupied by vulnerable people, a fact that has not contributed to produce spatial proximity between dissimilar socioeconomic groups. Consequently, the real estate market unlike Santiago, it has not worked to promote the spatial integration, but to consolidate economic differentiation in the urban space of intermediate cities. Summmarizing, since its foundation Temuco was considered as a political and institutional strategic point in order to give unity and spatial coherence to the Chilean State. Figure 6 shows the urban growth of Temuco since its origin until now. The city is characterized since its origins by its great economic, dynamic and demographic expansion, resulting from its great attractiveness and its status as regional capital, which influenced the arrival of investors and people mostly from its rural hinterland, looking for better living conditions. These factors played an important role to help consolidate this intermediate city in the Small South of Chile. However, the urban development process was not properly planned due to a number of deficiencies: predominance of reactive planning, uncompleted or non-updated instruments and non-efficient urban management, factors which influence the currently shape of the city. In the previous section, we revised how the urban segregation and fragmentation dynamics triggered by neoliberal model have introduced in the cities a series of challenges for policymakers and authorities. During the last decades new problems for the cities have become evident: economic standstill, poverty and pollution, these have also meant a drop in migration attraction and as a consequence a loss in the potential growth of the city which is evident in the inter-census period These are signs of the lack of urban sustainability of Temuco, according to our research. Some of the underlying root causes that may explain the problems the city is facing are derived from the regional context. The Araucania Region shows in the past decade an important backwardness of its economic base. This situation could be explained by the limited direct foreign investments during the past ten years Figure 7. The regional figures have an impact in the growth patterns of the city. Desarrollo inmobiliario: el nuevo rostro de Antofagasta Report No. In the same line, indicators such as labor market participation and unemployment rate within the Region also shows a negative situation, resulting in a direct impact on city growth. In La Araucania reached Conversely, in Chile poverty is defined taking as a reference the capability of a person to access to a basic food basket, which value is updated according to the evolution of prices Casen, Casen. Santiago: Ministerio de Desarrollo Social. These amounts were calculated based on the currency from January 17th, In the case of Temuco, This percentage is over the national average, which reaches This condition has been noticed by the local authorities, who are developing many initiatives in order to expand and improve the employability and also to develop programs to reinforce economic and productive projects, most of them to strengthen the capacities of small entrepreneurships Municipalidad de Temuco, Municipalidad de Temuco. Plan de desarrollo comunal. Both the negative and fragile economic conditions and the poverty situation that in general this region shows are key factors threatening the economic development and the possibilities of Temuco to consolidate itself as a crucial urban center in South Chile. The rapid urban growth experienced in Temuco between and has also meant an exacerbation of some environmental problems. One of these critical situations is the rise of air pollution. There are several reasons for this situation. Sanhueza et al. Impact of air pollution by fine particulate matter PM10 on daily mortality in Temuco, Chile. Revista Medica de Chile, 6 , Added to this, the situation becomes more critical with the use of wet wood in winter season, which is cheaper and easy to find. In fact, Sanhueza et al. Particulate air pollution and health effects for cardiovascular and respiratory causes in Temuco City, Chile. A second cause could be related to the progressive increase of the motorization rate, which is high correlated to the household income and the density of the city UDP, Universidad Diego Portales — UDP. Both factors are responsible for several episodes of environmental emergency along the year. In Temuco was declared as saturated zone by fine particulate matter PPM 2. Another important aspect to improve quality of life of the population which is in direct relation with sustainable development is related to infrastructure and the access to urban green areas. Public spaces and green areas provide social and ecological functions within urban environments Maas et al. Social contacts as a mechanism behind the relation between green space and health. In this sense, the lack of adequate public spaces and green areas in the city are evidenced also a critical situation. Urban planning, environment and health: from evidence to policy action. Geneva: WHO. We believe that as much as the regional economic regional standstill as the high poverty level and environmental pollution, all of them shown in Temuco most of all in the last millennium, are factors with a direct influence in the loss of migratory attraction to the city at a national level. The most direct consequence of this phenomenon is the deceleration of population growth that the city had had during the nineties Figure 8 , nevertheless, we believe that if the conditions continue to be the same there is a trend marked by the socio-environmental lack of sustainability. Inter and intra urban disparities, socio-spatial segregation, inequality and inadequate access to public services are now phenomena not restricted to big cities. This kind of generalization based on Borsdorf Borsdorf, A. Latin American urban development model is a preliminary and useful attempt to demonstrate that the dynamics of urban structuring in intermediate cities are comparable to major cities and the globalization effects are also impacting their urban development. Gated communities, pleasure lots in the fringe, and also an early but strong fragmentation represented by the spatial separation of different social groups in the city according to income differences are visible phenomena. However, in the case of Temuco there are some differences. First, the growth pattern associated to middle-to upper and upper classes neighborhoods is oriented to the west 2 2 For the case of Santiago, this pattern follows the opposite way. High income municipalities are located in the east part of the city. More information in Borsdorf The expansion of new urban structures, such as the Parcelas de agrado leisure plots by high income groups in peri-urban areas is also a remarkable aspect of this process. Secondly, although most of the commercial activities and services are concentrate mostly in the central business district CBD , in the past ten years they have experienced an expansion towards the Alemania avenue, also in the west part of the city. Thirdly, the speed of these changes is still slower than in metropolitan areas. Arenas, R. Coll Eds. This new drawing of the city left plenty of open questions and challenges that Temuco as intermediate city should be face in order to achieve a gravitating role in Chilean urban system and also a more sustainable future. One of the most relevant challenges is related to strengthen its economic function. National urban policy reviews: the case of Chile. Paris: OECD. In the Araucania Region, currently socioeconomic dynamics and the unresolved historical conflicts are responsible for the emerging of a loss of attractiveness for business and for these to attract migrant workers. These situations are influencing the currently economic situation of Temuco and its role in Chilean urban system. This scenario could be different if the dynamism that most of the medium and small cities showed in the past decade were seriously taking into account by policy makers and urban policies were capable to consider that physical capital, infrastructure and innovation are growth determinants that can enhance economic performances in these cities; and therefore not focuses almost all the investments plans exclusively in Santiago and the other metropolitan areas. The higher and not stopped inequality and poverty rate evidenced in Temuco are consequences of the centralized Chilean urban policy. Consider these ideas in currently discussions of how to improve sectorial and urban development policies could be one measure in order to improve competitiveness of intermediate cities. Secondly, knowledge about urban changes in intermediate cities should be the foundation on which new strategies for urban planning are implemented, seeking an answer, ideally with a decentralized approach to the local needs and the strains that put at risk sustainability in each of them. Open menu Brazil. Open menu. Abstract Resumo English Resumo Portuguese. Text EN Text English. AO07 copy. Abstract Academic production about the main Chilean cities exceeds the existing documentation on intermediate cities, though they have shown interesting trends patterns in recent years which have changed the urban system in Chile. Keywords: Urban development; Intermediate cities; Temuco. Introduction Most of urban literature shows that research has been focused on big metropolitan areas. Figure 3 - Nowadays, the Alemania Avenue is an emerging commercial and services hub in the city. Figure 4 - Evolution of the approved surface of building permits for residential uses between and Figure 5 - Gated communities in the peri-urban area of Temuco. Figure 6 - Urban growth of Temuco since its origins until now. For the case of Santiago, this pattern follows the opposite way. More information in Borsdorf Borsdorf, A. References Aylwin, J. Bolay, J. Borsdorf, A. Cabrales, L. Capel, H. Concha, C. Contreras, Y. Coy, M. De Mattos, C. Haller, A. Hidalgo, R. Hiernaux, D. Santiago: INE. Janoschka, M. Kaztman, R. Maas, J. Memoria Chilena. Banco de Chile. Mertins, G. Ministerio de Vivienda y Urbanismo. Municipalidad de Temuco. Navarrete, G. Observatorio Habitacional Minvu. Parnreiter, C. Reyes, S. Romero, H. Rovira, A. Sabatini, F. Sanhueza, P. Stadel, C. Toledo, X. Landsat Image. Vergara, L. History Received 13 May Accepted 07 Dec CMS is geographer; Dr. Figures 9. Source: Memoria Chilena Memoria Chilena. Source: Elaborated by the authors Stay informed of issues for this journal through your RSS reader. PDF English. Google Google Scholar. Urban growth trends in midsize Chilean cities: the case of Temuco.

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Official websites use. Share sensitive information only on official, secure websites. Cannabis plants and their seed have been used in many cultures as a source of medicine and feeding during history. Today, there is an increasing demand for cannabis seeds for medical use. Moreover, a seed sales market with no legal regulations has also grown. This may pose some issues if a quality control is not set in place. Identification of cannabis strains is important for quality control purposes in a nonregulated growing market and in cases of illegal traffic and medical use. Owing to the high price as a pharmacological drug, commercial products of cannabis plants and seeds for medical users are often subjected to adulterations, either when packing or distributing certified seeds in the market. Cannabis commercial seeds and cannabis seeds for medical use were analyzed with high-resolution melting HRM analysis using barcoding markers. Humulus lupulus L. DNA barcoding uses specific regions of the genome to identify differences in the genetic sequence of conserved regions such as internal transcribed spacer ITS and rbc L. DNA barcoding data can be generated with real-time polymerase chain reaction combined with HRM analysis to distinguish specific conserved DNA regions of closely related species. HRM analysis is the method of choice for rapid analysis of sequence variation. The melting temperature T m of homogeneous packages was consistent with single genotypes. However, packages containing contaminating seeds showed T m differences of 0. An effective, rapid, and low-cost method based on ITS nuclear DNA and on chloroplast rbc L regions for screening and detection of contamination in commercial cannabis seeds was developed and applied for the analysis of different samples. This approach can be used as a quality control tool for cannabis seeds or other plant material. The medical properties of Cannabis species have been known for centuries, but the interest on active secondary metabolites as alternative therapies for diverse pathologies has grown in recent years. This potential use has generated a strong impact worldwide concerning public health and the commerce of cannabis products. Indeed, this has led to legislation changes in some countries. Genetic identification of different cannabis strains is critical not only for quality control purposes in a nonregulated growing market, but for forensic purposes and for medical use as well. Its medical use has been focused on the treatment of skin pathologies, seizures, lack of appetite, neurodegenerative diseases, multiple sclerosis, as well as for the management of chemotherapy side effects 3—6 and chronic pain treatment. This could produce cognitive disorders related to day-to-day consumption and especially in cases of long-term cannabis use. This poses a problem since these plant products require a certificate of authentication. Various methods of identification have been developed to authenticate medical plant species such as morphological analysis, chemical profiling, and DNA-based molecular analysis. The study of conserved regions of the genome for identification has allowed broadening the spectrum of analysis in a number of species. This has been applied on both the botanic and forensic fields. These molecular methods can detect differences at the DNA level and they offer numerous advantages over the conventional phenotype-based approaches. The ITS2 region is one of the most used universal barcode for plant species identification. HRM analysis is the method of choice for rapid analysis of sequence variation within PCR amplicons by determining their melting temperature T m. T m differences can be detected by monitoring the fluorescence changes during PCR cycles using real-time PCR differential analysis. Genotypes are then differentiated by their characteristic melting curves, visualized by the loss of fluorescence as the DNA duplex melts 23 HRM analysis allows genotyping of plant, fungus, and animal species. DNA sequence differences, such as single nucleotide polymorphisms and small insertions and deletions Indels , can be detected based on the location of a differential peak and the shape of the melting transition curves. This article describes a case of adulteration of cannabis seeds due to lack of control during the packaging process. Cannabis sativa L. The objective of this study was to authenticate cannabis strains using a real-time PCR and HRM-based molecular method. Using ITS2 and rbcL barcoding HRM, it was possible to detect the presence of different strains in commercial cannabis seed used for medicinal purposes. This method can be very valuable in terms of traceability and authentication of strains in cannabis commercial seeds. Commercial seeds of four different C. Commercial Hops Humulus lupulus L. Plant material was previously powderized before extraction. DNA was quantified using a fluorometer Qubit 2. Genomic DNA integrity was confirmed by agarose gel electrophoresis. Amplicon size and primer specificity were confirmed by 1. The fluorescent data were acquired at the end of each increment step. All experiments were run in duplicate. Melting curve analysis was performed with the Rotor-Gene Q software v. The first step was to calculate triangular similarity matrix based on Euclidean distance and then a multidimensional metric scaling mMDS was performed. This analysis determines the relationships between the profiles at different temperatures for each marker. A k-means clustering was used to detect classes through a set of quantitative variables. DNA extracted from all seeds yielded a specific amplification product for both regions. These sizes are in accordance to previously published sequence data. PCR amplicons were analyzed to determine their specific T m. Results are represented by means of conventional derivative plots. The melting curve is generated by slowly melting the DNA through a range of temperatures in the presence of a dsDNA binding dye. Distinguishable melting curves were obtained for H. T m s resulted to be All cannabis seeds provided by customers yielded different melting curves. A Melting curve profile using ITS region. C Melting curve profile using rbcL marker. The analysis of ITS2 region yielded different outcomes for the four analyzed strains. Two of the strains resulted to have the same genetic profile Fig. A Melting curve profile of three cannabis seeds with same sequence. B Normalized curve profile of three cannabis seeds with same sequence. C Melting curve profile of two cannabis seeds with same sequence and one seed with different sequence. D Normalized curve profile of two cannabis seeds with same sequence and one seed with different sequence. These differences depend on GC content, length of amplified product, and sequence. The undistinguishable seeds resulted to have a consistent Tm of ITS2 results were confirmed through rbcL analysis. Analysis of derivative melt curve resulted in consistent T m s, within the range Three cannabis seeds resulted to have the same rbc L sequence Fig. HRM analysis using the rbc L chloroplast marker. The mMDS analysis showed the relationships between the seeds' profiles at different temperatures for each gene Fig. Four sets of three seeds from different strains were included in the analysis. All seeds from sets 2 and 4 resulted to form a cluster, whereas seeds from sets 1 and 3 appeared to be genetically heterogeneous. A mMDS represent seed samples of the four different packages used in the study. Each point represents a seed for ITS region. B mMDS represent seed samples of the four different packages used in the study. Each point represents a seed for rbc L marker. However, samples from sets 1 and 3 were not able to be assigned to the same cluster due to their genetic sequence difference. These results demonstrated that seeds from sets 2 and 4 appeared to be genetically homogeneous, whereas samples from sets 1 and 3 seemed to be genetically heterogeneous. Results from sets 1 and 3 clearly indicated the presence of a mixture of seeds. We described a case in which 10 samples of seeds from medicinal users and four strains of cannabis seeds were analyzed by HRM coupled with barcoding. This authentication is critical to ensuring the quality of medical cannabis seed. However, up to date there is no standardized method for rapid identification of cannabis. DNA sequencing technology is relatively expensive and time consuming. Thus, Bar-HRM is a better option for a rapid screening of cannabis strains. However, HRM shows some limitations when the genetic variation is extremely small. Four strains were purchased from a specialized store that distributes cannabis seeds for recreational and medicinal use. Molecular authentication showed that cannabis seed are good sources of DNA, providing full and amplifiable genetic material that can be used efficiently for authentication analysis by HRM. Distinction down to genus and—in many cases—species level is possible based on melting temperatures T m of specific PCR products. The ITS2 region was selected as a barcode marker because is widely used for phylogenetic reconstructions at both genus and species levels. The use of a combination of barcode regions is common and recommended for plant identification. To confirm that each package contained seeds of the same strain, we decided to analyze randomly purchased commercial products. In some cases, some cannabis users reported that different phenotypes were observed from seeds purchased as belonging to a single strain. The robustness of such analyzes would also be strengthened by the exploration of markers in organelles and by increasing the universe of samples to be incorporated in the studies. Our results proved that seeds had different genetic sequences. This possible adulteration may be attributed to poor packaging or a mistake during package processing. Each package of each strain contains three seeds each. Surprisingly, it was found that two out of four packages contained mixtures of seeds. We proposed a fast easy low-cost method to distinguish the presence of mixtures in seed packages. This can certainly help confirm the authenticity or at least the homogeneity of the seed contents of a given strain. Although this analysis can be done at any stage of plant development, it was demonstrated that seeds were a good source of amplifiable DNA. The homogeneous packages had the same T m for each of the three seeds of that strain. In contrast, packages containing mixed seeds displayed different T m s. This allowed distinguishing the different genotypes in the same package. In this study, Bar-HRM analysis was proposed to be a fast and accurate technique for authentication testing of cannabis seed of commercial strains. Here, we describe the development of a Bar-HRM method for adulteration testing of cannabis seeds contained in individual packages. These packages correspond to different cannabis strains. This method allowed detecting heterogeneity in two out of four cases in the contents of these packages. This is undoubtedly important at the time of consumption of these plants by medicinal users. In future studies, we will continue to explore the different seed banks commonly used by users of cannabis and incorporating new DNA markers. We wish to confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this study that could have influenced the results. This study was partially funded by EM-LA The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the article. As a library, NLM provides access to scientific literature. Cannabis Cannabinoid Res. Find articles by Jaime Solano. Find articles by Francisco Encina-Montoya. Find articles by Marco Bustos. Find articles by Alejandra Figueroa. Find articles by David Gangitano. Collection date Aug. Copyright , Mary Ann Liebert, Inc. Open in a new tab. Similar articles. Add to Collections. Create a new collection. Add to an existing collection. Choose a collection Unable to load your collection due to an error Please try again. Add Cancel.

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