Andean farmers of South America took the first steps in domesticating quinoa from its wild forms more than 5,000 years ago (Planella et al., 2015). Seed exchanges and migrations of human populations played an important role in the early development of landraces with indehiscent, larger seeds, adaptations to drought and saline soils, and varietal differences in photoperiod, altitude, and diverse rainfall conditions (Bazile et al., 2013). Five ecotypes are globally recognized: Inter-Andean valleys (Colombia, Ecuador, and Peru); Altiplano (northern highlands in Peru and Bolivia); Yunga (Bolivia); Salares (salt flats or southern highlands in Bolivia, Chile and Argentina); and Coastal (coastal or sea level areas in central and southern Chile; Risi and Galwey, 1984; Fuentes et al., 2012; Bazile et al., 2013). Long-term selection in these sub-centers of quinoa biodiversity confers specific and often unique traits of adaptation to the landraces. Diversity is observed in a wide array of colors of plants and seeds, and through differences in branching patterns and panicle types, grain productivity, abiotic stress tolerances and disease resistance (Fuentes and Bhargava, 2011; Ruiz-Carrasco et al., 2011). Even if the potential adaptation to climate change inherent in quinoa confers probable utilization for agricultural development in other parts of the world, a better understanding of the diversity of the actual contexts of cultivation in its area of origin is crucial in order to predict where specific quinoa ecotypes would have the most agronomic impact (Jacobsen et al., 2015). Quinoa has higher drought and salinity tolerance than many other crops, and tremendous variation in these traits are evident among quinoa ecotypes and existing quinoa varieties and landraces (Bazile et al., 2015; Peterson and Murphy, 2015a). For example, the Salares ecotype is best suited to withstand extreme conditions of this cold high-altitude desert at 4.000 m.a.s.l. This ecotype is extremely drought resistant and adapted to saline and sandy soils surrounding the salt lakes. Alternatively, the quinoa of the Coastal ecotype grows from sea level up to 1500 m.a.s.l. and possesses characteristics specific to this unique environment; specifically, adaptation to high annual precipitation, resistant to pre-harvest sprouting, high evapotranspiration index, a comparatively high degree of heat tolerance, and salinity tolerance.

Due to quinoa's increasing popularity and recent expansion of cultivated area on a global scale, farmers and plant scientists are emphasizing morphological and physiological modifications that allow for mechanized harvesting, improved heat tolerance and downy mildew resistance, and reduced agricultural inputs (Bazile et al., 2015; Bonifacio et al., 2015). New sources of heat tolerance or disease resistance genes from wild relatives are being introgressed into commercial varieties by plant breeders; however, undesirable characteristics such as seed dormancy and shattering from the donor species can decelerate a quinoabreeding program (Zurita-Silva et al., 2014). The objective of this long-term participatory development project is to implement and successfully conduct a collaborative European-North American EPB program for quinoa with field sites in the host countries and in South America as well as across several countries in Africa and Asia.

### PARTICIPATORY PLANT BREEDING

Participatory variety selection (PVS) and Participatory plant breeding (PPB) have been practiced in various regions and on various crops over the last 30 or more years. Much of the focus has been in developing countries, but there has also been some work in developed countries, often with a focus on low-input and/or organic agriculture (Chable et al., 2008; Ghaouti et al., 2008; Leroy et al., 2014; Campanelli et al., 2015). The primary difference between PPB and PVS is the extent of farmer participation in the various stages of the breeding program. PVS involves testing and selecting new varieties developed by the institutional system within farmers' fields and at local research stations in various environments and allowing farmers to compare these varieties with local farmer varieties. In a PPB program, farmers are routinely involved in decision-making throughout the entire breeding process and not just in the final testing of advanced breeding lines. Trials are conducted in farmers' fields and managed by farmers. Farmers score varieties according to criteria that they define. Data generated by breeders provides the basis for the selection of successful varieties by farmers, in dialog with breeders.

On marginal lands, and in relation to NUS and noncommercial crops, formal plant breeding has made less of an impact (Ceccarelli et al., 2009). To address this, plant breeders working on a diversity of crops in various regions began involving farmers in setting breeding objectives and in the subsequent selection and testing of breeding materials. This direct dialog benefits the breeding process with possible farmer participation in setting objectives, creating variability, selecting and testing breeding material, and producing and diffusing seed of new cultivars (Weltzien and Christinck, 2008). The objective of PPB research was to more appropriately respond to the specific needs of under-represented farmers and to optimize production, adoption, and food sovereignty through improved local adaptation of cultivars to relevant abiotic and biotic pressures, as well as to prevalent cultural norms (Ceccarelli et al., 2001; Sperling et al., 2001; Weltzien et al., 2003). Improvement of PPB continues as researchers identify different factors that affect breeding material performance including number of entries to be evaluated (Thiele et al., 1997; Ceccarelli et al., 2000), gender (Kamara et al., 1996; Sthapit et al., 1996; Weltzien et al., 1996), independent versus paired or group evaluations, and inclusion of on-station evaluations by farmers (McElhinny et al., 2007).

Among the important outcomes of PPB is the positive impact it has on biodiversity. PPB is a highly decentralized process where research occurs in a multitude of farmers' fields in different ecoregions and microclimates. PPB produces resilient varieties with high levels of phenotypic plasticity that often differ across locations depending on local climate and ecosystems, as well as social, cultural and economic factors influencing farmer

preferences. With regard to quinoa, this diversity can be easily observed in situ in farmers' fields showing a wide array of colors in plants and seeds, different types of branching and panicles, as well as having variation in grain productivity, abiotic stress tolerance and disease resistance (Bazile et al., 2013). Varieties selected by farmers are often genetically variable, similar to traditional farmer landraces, in stark contrast to the majority of varieties produced by conventional breeding in which all the plants are genetically identical. The fact that farmers' varieties are not genetically uniform is precisely what makes them resilient to a variety of stresses that are made more unpredictable by climate change.

### EVOLUTIONARY PARTICIPATORY BREEDING (EPB)

Due to the predominance of monocultures in global crop production, crop diversity occurs at a scale where individual genotype unit areas (GUAs; Mundt and Browning, 1985) are often many square kilometers (Newton et al., 2009). The genetic uniformity inherent in monocultures can restrict the crop's ability to tolerate diverse abiotic environmental stresses, pests and diseases, thereby leading to a potential decrease in the stability of the cropping system (Hooper et al., 2005; Hughes et al., 2008). Chakraborty and Newton (2011) suggest that strategies for establishing greater resilience and improved yield stability should focus on the introduction of increased genetic variability, both within and between cultivars, into agricultural systems. Such crop populations are better equipped to adapt to future unpredictable temporal climate shifts than are monocultures (Louafi et al., 2013). Genetically uniform cultivars have been shown to lack the ability to adjust and adapt to highly unpredictable environmental fluctuations, in direction and range, and novel stress factors (Verboom et al., 2010). Evolutionary breeding provides a framework with which to address current and predicted threats to agriculture as a result of climate change and agricultural intensification.

Evolutionary breeding involves four distinct stages: (i) creation of genetic diversity; (ii) multiplication of seeds from each cross followed by an equal mixing of populations if more than one cross will be used in a given population; (iii) repeated sowing and harvesting of the population in one or more agronomic environment without active selection of individual plants; and (iv) use of the seed in culturally relevant settings as food or feed, or use of the seed as a basis for further breeding (Döring et al., 2011). EPB merges the evolutionary breeding method described by Suneson (1956), Allard and Adams (1969), Allard (1988) and Phillips and Wolfe (2005) with farmer participatory breeding to develop high-yielding, disease-resistant cultivars of desired quality while maintaining a high degree of genetic variation to allow for adaptability to fluctuations in environmental conditions (Murphy et al., 2005; Döring et al., 2011; Murphy et al., 2013; Ruiz et al., 2014; Bazile, 2015).

### QUINOA POPULATIONS AND PRELIMINARY EPB METHODOLOGY

Ten biparental quinoa populations were developed at Washington State University using seed from the USDA National Genetic Resources Program and from private seed companies in the US (**Table 1**) (Peterson et al., 2015). Parents were chosen based on one or more of the following characteristics from results of several years of preliminary testing: seed yield, day-neutral photoperiod, resistance to downy mildew, heat tolerance, drought tolerance, seed size, plant height, tolerance to lodging, mold and pre-harvest sprouting resistance, and plant and seed color. Seed was advanced to the F3 and populations were planted on two organic farms, located in Pullman and Quilcene, in 2014, each representing a distinct location and associated agronomic challenges (Washington State, USA). Pullman represents a very hot and dry climate in the major wheat growing region of the Pacific Northwest. Quilcene represents a cool, coastal climate with where late season rains caused significant yield loss due to pre-harvest sprouting and downy mildew (Peronospora variabilis) in 2010 and 2013.

No human selection was conducted on the F3 quinoa populations in 2014, the hottest year on record in eastern Washington. Extremely low population yields in Pullman reflect

TABLE 1 | Pedigree and seed yield of 10 populations in two locations (Pullman: High-heat environment and Quilcene: Low-heat environment) in Washington State, USA in 2014.

the effect of heat on pollen viability (**Table 1**). Variety trials, which included the majority the parents of the populations, adjacent to the population trials yielded no seed at all in any of the varieties. Population 102, derived from two relatively heat tolerant parents (Colorado 407D and QQ74) showed the highest seed yield (**Table 1**). The populations in Quilcene were not subject to heat stress, nor stress due to excessive rainfall, and subsequently seed yield was much higher. This evolutionary breeding method was repeated with F4 seed in 2015 in Quilcene, however, instead of a bulk harvest, 200 panicles from each population were harvested individually after a deliberate and prolonged exposure to rainy early autumn conditions to encourage pre-harvest sprouting in the populations. Additionally, farmers from a range of farms in the Olympic Peninsula conducted both negative and positive selection on six populations on a nearby organic farm in Chimacum, Washington. Some individual seed from each positively panicle will be saved and replanted in pedigree headrows in 2016, but the majority of the F5 seed will be bulked and the populations reconstituted; this seed will be distributed to collaborators in partner countries to begin the EPB process in 2016.

### PERSPECTIVES ON A GCN-QUINOA

The pilot GCN-Quinoa is the first step in moving forward with EPB in quinoa. After just one year of existence, more than 150 researchers are part of this global network and represent a diversity of people from 60 countries who currently share a similar curiosity in adapting quinoa to their region (**Figure 1**). Nowadays, the number of research centers with experimental quinoa programs is increasing worldwide, even in regions that are not yet producing or consuming quinoa (Bazile et al., 2015; Peterson and Murphy, 2015b). However, the primary quinoa research centers are located in countries where quinoa has been grown traditionally for centuries. As a result, there exists a significant gap in varietal, agronomic, and cultural knowledge of quinoa production and consumption in Africa for example. In that context, the main objective of the GCN-Quinoa is to create a collaborative space to facilitate exchanges between producers, processors, distributors, politicians, and all persons involved in quinoa development for the promotion and sustainable use of quinoa genetic resources. Access and benefit sharing about quinoa seeds are of importance that is why a specific forum of discussion was generated online on this topic. Members of the GCN-quinoa interested in testing quinoa through EPB approach have joined, and can continue to join, the development of the Worldwide Consortium on Evolutionary Participatory Breeding in Quinoa and benefit from the expertise and support of the group.

Increasing consumer demand in quinoa and the usefulness of the crop in adapting to climate change means that the cultivation of quinoa is undergoing rapid expansion worldwide. It thus acknowledges the role that quinoa's biodiversity and high

nutritional value could play in providing global food security. Since this minor crop could become a major crop, how can we continue the varietal improvement efforts that farmers in the Andean countries have been carrying out for generations? The achievement of this goal requires innovative approaches to plant breeding in order to avoid the well-known risks of genetic uniformity and vulnerability as well as to encourage sustainable agriculture with a decreased use of agrochemical inputs.

How does one breed for farmer's needs? Involving a large number of farmers in evaluation and selection throughout the selection process is an excellent way to take into account heterogeneity across environments (Kotschi, 2010). This form of decentralized selection is critical in involving and empowering farmers from the beginning of the breeding process. There is a need for a paradigm shift in agriculture which must enhance the role of agrobiodiversity and the need for innovation in plant breeding is particularly crucial (Ceccarelli, 2009). Increasing genetic diversity in quinoa populations may be a relevant method to adapt crops to environmental changes, in particular, to agricultural areas where environmental conditions are marginal for crop production. It will be important to consider market options, and the desire among end-users for a uniform product, when developing populations. Therefore, particular attention must be given to the selection toward uniformity of targeted quality traits, such as seed size and color, which can be identified visually post-harvest, and protein content and starch characteristics, which can be evaluated in a lab (Wu et al., 2014).

Evolutionary PPB with quinoa has the potential to contribute significantly to sustainable agriculture if a lasting connection can be made among all the relevant stakeholders. Linking researchers at the global level who are involved with quinoa trials in a collaborative network may be the most effective pathway. One of the challenges offered during the (IYQ, 2013) was to devise a means and methodology to increase the likelihood of quinoa's success in the introduction and development in non-traditional quinoa growing regions. Next-generation quinoa research will require a collaborative effort from a dedicated group of experts who possess the ability to evaluate the diverse global needs of quinoa research and to subsequently conduct this research in a cooperative manner (Bazile, 2013).

Given that quinoa cultivation and research is expanding to parts of world that are not familiar with the crop, researchers and farmers would benefit from being included in a network with other researchers facing similar challenges. They would also benefit from access to a wide range of germplasm which is currently concentrated in gene banks in Latin America, as noted above, and to which researchers in other parts of the world may not have ready access (Bazile et al., 2016). The International Treaty on Plant Genetic Resources for Food and Agriculture (the Treaty) proposes the implementation of a multilateral system (MLS) of exchange of genetic resources for both facilitate access to genetic resources and ensure benefit-sharing. The Treaty provides a monitoring of genetic resources with the harmonized material transfer agreements; certification of origin to prevent biopiracy and the disclosure of origin to share the benefits. With few notable exceptions (Argentina, Bolivia, and Colombia), Andean countries1 have ratified the Treaty. Under its MLS, countries agree to make available their genetic resources stored in collections that are under their control and in the public domain for 64 crops that are listed in Annex 1 of the Treaty. Although some discussions are under way to possibly enlarge this list, quinoa does not currently belong to this Annex 1 which means that, so far, the rules of the MLS do not apply for the exchanges of quinoa genetic material. The Convention on Biological Diversity (CBD) does only regulate bilateral access and benefit sharing, but this is difficult to apply to quinoa as the crop is now planted internationally, not restricted to the Andean region, and this has been the case for decades. For quinoa, there is no single existing legal framework providing a comprehensive coverage of all the issues related to the genetic resources and their sustainable management (Chevarria-Lazo et al., 2015). Considering that the status of quinoa is now rapidly changing from a NUS to a major crop for food and agriculture, the opportunity exists to question current dominant agricultural models and plant breeding methods and begin to move forward toward more participatory approaches in order to address the real needs of the end-users in a context of global changes (Anvar, 2008; Trommetter, 2010). Research and plant breeding in quinoa should be based on more flexible genetic material with the potential to maintain yield stability while continually evolving in response to changes in climate. Considering the GCN-Quinoa as a vector to disseminate evolutionary material, considered as open seed sources, with a high level of genetic diversity will certainly favor adaptation in new environments. Nevertheless, without direct dialog resulting from close collaborations between farmers and researchers, we may struggle to reach the next step for quinoa introduction to cropping systems for local diets.

### AUTHOR CONTRIBUTIONS

KM wrote two sections of, and edited the entire manuscript. KM initiated the quinoa populations and along with JK set up and conducted the participatory breeding research. JK provided edits to the entire manuscript. DB and MR contributed significantly to the writing and editing of the manuscript. In addition, DB. initiated the Global Collaborative Network on Quinoa.

### ACKNOWLEDGMENT

The authors thank all of the researchers, farmers and stakeholders who have already manifested their interest in joining the pilot GCN-Quinoa (gcn-quinoa.org) and who have requested quinoa crop populations to initiate trials.

1 Contracting Parties: Chile (2016), Ecuador (2004), Peru (2003), Venezuela (2005); Only Signatorie: Argentina (2002), Colombia (2002); Non Contracting parties: Bolivia.

### REFERENCES

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security under climate change. A review. Agron. Sustain. Dev. 34, 349–359. doi: 10.1007/s13593-013-0195-0

**Conflict of Interest Statement:** The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

The reviewer RP and handling Editor declared their shared affiliation, and the handling Editor states that the process nevertheless met the standards of a fair and objective review.

Copyright © 2016 Murphy, Bazile, Kellogg and Rahmanian. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

# Worldwide Evaluations of Quinoa: Preliminary Results from Post International Year of Quinoa FAO Projects in Nine Countries

Didier Bazile1 \*, Cataldo Pulvento2 , Alexis Verniau3 , Mohammad S. Al-Nusairi 4 , Djibi Ba5 , Joelle Breidy 6 , Layth Hassan7 , Maarouf I. Mohammed8 , Omurbek Mambetov 9 , Munira Otambekova10, Niaz Ali Sepahvand11, Amr Shams 12, Djamel Souici 13, Khaled Miri 11 and Stefano Padulosi 14

#### Edited by:

Soren K. Rasmussen, University of Copenhagen, Denmark

#### Reviewed by:

Michael Benjamin Kantar, University of Hawaii, USA Redouane Choukallah, International Center for Biosaline Agriculture (ICBA), United Arab Emirates

> \*Correspondence: Didier Bazile<EMAIL_ADDRESS>
#### Specialty section:

This article was submitted to Crop Science and Horticulture, a section of the journal Frontiers in Plant Science

> Received: 24 March 2016 Accepted: 30 May 2016 Published: 21 June 2016

#### Citation:

Bazile D, Pulvento C, Verniau A, Al-Nusairi MS, Ba D, Breidy J, Hassan L, Mohammed MI, Mambetov O, Otambekova M, Sepahvand NA, Shams A, Souici D, Miri K and Padulosi S (2016) Worldwide Evaluations of Quinoa: Preliminary Results from Post International Year of Quinoa FAO Projects in Nine Countries. Front. Plant Sci. 7:850. doi: 10.3389/fpls.2016.00850 1 Unité Propre de Recherche Gestion des Ressources Renouvelables et Environnement, Department of Environments and Societies, French Agricultural Research and International Cooperation Organization, Montpellier, France, 2 CNR–Institute for Agricultural and Forest Systems in the Mediterranean, Ercolano, Italy, 3 ESA–Ecole Supérieur d'Agriculture, Angers, France, 4 General Seed Multiplication Cooperation, Agricultural Research and Extension Authority, Dhamar, Yemen, 5 Centre National de Recherche Agronomique et de Développement Agricole, Kaedi, Mauritania, 6 Department of Plant Breeding, Lebanese Agricultural Research Institute, Zahlé, Lebanon, 7 Plant Production Department, Ministry of Agriculture, Baghdad, Iraq, 8 Genetics and Plant Breeding, Forage and Range Research Program, Agricultural Research Corporation, Khartoum North, Sudan, 9 Seed Association of Kyrgyzstan, Bishkek, Kyrgyz, 10 Seed Association of Tajikistan, Dushanbe, Tajikistan, 11 Research Department, Seed and Plant Improvement Institute, Karaj, Iran, 12 Crop Intensification Research Department, Field Crops Research Institute, Agricultural Research Center, Giza, Egypt, 13 Département Recherche Expérimentation, Institut Technique de Développement de l'Agronomie Saharienne, Biskra, Algeria, 14 Bioversity International, Maccarese, Italy

Chenopodium quinoa Willd., a high quality grain crop, is resistant to abiotic stresses (drought, cold, and salt) and offers an optimal source of protein. Quinoa represents a symbol of crop genetic diversity across the Andean region. In recent years, this crop has undergone a major expansion outside its countries of origin. The activities carried out within the framework of the International Year of Quinoa provided a great contribution to raise awareness on the multiple benefits of quinoa as well as to its wider cultivation at the global level. FAO is actively involved in promoting and evaluating the cultivation of quinoa in 26 countries outside the Andean region with the aim to strengthen food and nutrition security. The main goal of this research is to evaluate the adaptability of selected quinoa genotypes under different environments outside the Andean region. This paper presents the preliminary results from nine countries. Field evaluations were conducted during 2013/2014 and 2014/2015 in Asia (Kyrgyzstan and Tajikistan), and the Near East and North African countries (Algeria, Egypt, Iraq, Iran, Lebanon, Mauritania, and Yemen). In each country, the trials were carried out in different locations that globally represent the diversity of 19 agrarian systems under different agro-ecological conditions. Twenty-one genotypes of quinoa were tested using the same experimental protocol in all locations consisting in a randomized complete block design (RCBD) with three replicates. Some genotypes showed higher yields and the Q18 and Q12 landraces displayed greater adaptation than others to new environmental conditions. The Q21 and Q26 landraces were evaluated with stable and satisfactory levels of yield (>1 t.ha−1 ) in each of the different trial sites. This production stability is of considerable importance especially under climate change uncertainty. While these results suggest that this Andean crop is able to grow in many different environments, social, and cultural considerations remain crucial regarding its possible introduction as a staple food in new cropping systems around the world.

Keywords: Chenopodium quinoa Willd., plant genetic resources, seeds, adaptation, climate change, multi-local trials, agrobiodiversity, agroecology

### INTRODUCTION

Chenopodium quinoa Willd. is a plant is a plant originated in the Andean Plateau, around Lake Titicaca, 3800 m above sea level (m.a.s.l.) on the Peruvian-Bolivian border (Heiser and Nelson, 1974; Jacobsen, 2003). Quinoa domestication began there about 7000 years ago (Bazile et al., 2013), and brought a significant increase in the genetic diversity of the species cultivated (Bhargava et al., 2007a,b). Such a high genetic diversity in quinoa is closely related to the vastness of its center of origin and the varied of human uses that have been influencing the selection process over time (Bazile and Negrete, 2009). This is the case with many domestication processes of numerous important crops and their wild relatives (Wilson, 1990). The selection exerted by local communities has led to many landraces, which can be still found grown especially in Bolivia and Peru (Risi and Galwey, 1984; Bazile et al., 2014; Bazile, 2015).

At global level, there are more than 6000 varieties of quinoa cultivated by farmers (Rojas et al., 2015). Those varieties can be classified into five main categories or ecotypes, according to their adaptation to specific agro-ecological conditions in major production areas (Bois et al., 2006; Rojas, 2003; Anabalón and Thomet-Isla, 2009; Fuentes et al., 2009a; Fuentes and Bhargava, 2011; Bazile et al., 2013, 2014). Quinoa of the inter-Andean valleys grows in areas between 2300 and 3500 m.a.s.l., characterized by annual rainfall between 700 and 1500 mm. Quinoa grows in highlands (also called Altiplano of the Andes) between 3500 and 3900 m.a.s.l. in areas with an annual rainfall of 400–800 mm. Quinoa from the edges of deserts and high altitude salt lakes (Salares) grows in areas nearly 4000 m.a.s.l., characterized by a limited volume of annual rainfall (150–300 mm) and with many days of frost. Quinoa found at sea level (Coastal) is adapted to the regions lying between sea level and 1000 m.a.s.l., where annual rainfall ranges from 500 to 1500 mm (Martínez et al., 2009). Quinoa from the Yungas grows under tropical moisture conditions and in areas with high levels of precipitations.

Considering the high genetic diversity of quinoa, the needs of the crop vary extensively by landrace or cultivar (Cleveland et al., 1994; Brookfield et al., 2002; Chevassus-au-Louis and Bazile, 2008). Due to the diverse characteristics of the five ecotypes, quinoa can be grown under very different climatic conditions (Jacobsen, 1997; Jacobsen et al., 2003; Christensen et al., 2007; Fuentes et al., 2009b, 2012). Sandy loam soils with good drainage and if possible, a high content of organic matter and nutrients are preferable for quinoa to better adapt to new environments. It is also advisable to work in neutral soils, although quinoa can tolerate different pH and grow in alkaline (to pH 9) and acid soils as well (up to pH 4.5; Narea, 1976; Tapia, 1979). As previously described, quinoa cultivation can be pursued in many climatic conditions, including desert, hot, dry, cold and dry, temperate, and rainy or hot with high humidity (Bosque et al., 2003; Gesinski, 2008). Indeed, scientific evidence exist to confirm that quinoa tolerates very dry conditions and drought: quinoa uses water very effectively, even though it is a C3 plant, due to physiological mechanisms (Cocozza et al., 2013) that allow the plant to prevent moisture deficits, and tolerate and/or withstand lack of soil moisture. An ideal average temperature for quinoa would be around 15–20◦C, but some specific landraces can also withstand extreme temperatures from −8 ◦C to +38◦C (Bazile et al., 2015). Sensitivity periods to temperatures have been recorded mainly when seeds germination occurs in cold temperatures (frost) and when flowering takes place under high temperatures (FAO, 2011). Quinoa is able to endure extreme solar radiation, allowing it to store hours of heat necessary to carry out its vegetative and productive phases. Quinoa is cultivated in areas from 2◦ North latitude to 47◦ South latitude, from Colombia to Chile in South America.

There are varieties adapted to short days, long days and those photoperiod insensitive (Bertero et al., 1999; Bertero, 2001). There are two major phases in the development of quinoa plants: the vegetative phase is an active growth phase during which the plant acquires new properties to reach vegetative maturity. This is followed by the reproductive phase, which is the period when the plant will be able to produce flowers and seeds and reach physiological maturity. Depending on the photoperiod sensibility of each variety, each stage's duration can be modified depending on the length of days and on temperatures (Risi and Galwey, 1989, 1991; Jacobsen and Stølen, 1993). Photoperiod sensitivity is a key factor in the adaptation of this crop at new latitudes (Bertero, 2001).

For thousands of years, quinoa has been a staple food for Andean populations (Tagle and Planella, 2002; Planella et al., 2011; Martinez et al., 2015). Mainly used as a cereal grain (it is in many cases considered a pseudo-cereal), botanically speaking quinoa is an achene, a seed-like fruit with a hard coat (Cusack, 1984; NRC, 1989). Classified as a member of the Amaranthaceae (a large family of 160 genera and 2400 species- Sing, 2010) from the genus Chenopodium, the specie C. quinoa Willdenow is gaining importance particularly for its high and wellbalanced nutritional contents. Quinoa has exceptional nutritional properties, with high protein content in comparison to cereals, which is combined with a good balance of essential amino acids (Vega-Gálvez et al., 2010; Maureira and Martínez, 2012; Miranda et al., 2012; Lutz et al., 2013). The quinoa crop has recently undergone important developments around the world regarding its ability to withstand extreme conditions (Bazile et al., 2015). Its high genetic diversity provides opportunities for leveraging its hardiness and further its wide adaptation (Louafi et al., 2013). Many countries regularly face food insecurity problems, often combined with a difficult agricultural environment (FAO, 2015). Today, drought and soil salinization are major limiting factors in cultivation, a fact that is generating significant pressure on arable land availability. Considering these major challenges, quinoa hardiness is increasingly being appreciated by growers: today the crop is presently cultivated, or is under experimentation, in more than 95 countries and its cultivation continue to expand rapidly worldwide (Bazile, 2015; Bazile et al., 2016).

The main quinoa producers in the world are Bolivia, Peru, Ecuador, and the United States of America. In 2013, over 75,000 hectares of land were under quinoa cultivation in Bolivia and more than 45,000 hectares in Peru. These two countries are still the major producers in the Andes and in the world. Today the cultivation of quinoa has reached countries as far as Tibet, Morocco, France, India, China, the United Kingdom, Sweden, Denmark, Netherlands, and Italy, among others (Bhargava et al., 2006; Pulvento et al., 2010; Bazile, 2015; Bazile et al., 2015). From the 1950s to nowadays, trials for plant breeding and/or crop adaptation have been conducted in Andean countries but also in other parts of the world in order to better understand the domestication process of quinoa and to obtain quinoa germplasm adapted to new environmental conditions (Bonifacio et al., 2015; Jellen et al., 2015). Quinoa is a viable alternative for food insecure countries in a world facing increasingly climate challenges and set to feed a growing population in terms of both food and nutrition security (Galwey, 1992; Ruiz et al., 2014).

The FAO project "American and European Test of Quinoa" (1996–1998) was the first mechanism for the diffusion of quinoa worldwide and underlies the current global expansion of the crop (Mujica et al., 2001). Field trials were established in several countries to evaluate the performance of quinoa through multiple experiments at the international level. Since 1996, quinoa has been recognized as one of the most promising crops in terms of food security (Schlick and Bubenheim, 1996). While the major producers are still located in the Andean region, quinoa cultivation is a reality across all continents where germplasm originated from the Andes was successfully selected (Bazile et al., 2016).

The first objective of the International Year of Quinoa (IYQ) in 2013 was to increase the visibility of the great potential of quinoa biodiversity to contribute to global food security, especially in countries where the population has no access to other protein sources or where production conditions are limiting. In many of the countries in North Africa and the Near East, food security still remains a major problem for vulnerable population groups.  Its high tolerance to extreme weather conditions and high nutritional value contributed to choosing quinoa as a potential crop to address these challenges (Delatorre-Herrera and Pinto, 2009; Orsini et al., 2011; Ruiz-Carrasco et al., 2011; Pulvento et al., 2012). Following the IYQ, the Food and Agriculture Organization of the United Nations (FAO) initiated a multi local test (through FAO-TCP for Technical Cooperation Programmes) in a number of countries of North Africa, the Near East and Asia viz. Algeria, Egypt, Iran, Iraq, Kyrgyzstan, Lebanon, Mauritania, Sudan, Tajikistan, and Yemen. During the implementation of these projects, FAO collaborated with many partners worldwide to access quinoa seeds and their expertise in the field of crop cultivation. Various research centers, universities and seed firms were mobilized to find quinoa seeds of different varieties to be tested. Twenty different genotypes were eventually made available to countries for the international trial programme.

The main goal of this research was to evaluate the adaptability of selected quinoa genotypes under different environments outside the Andean region. Two hypotheses have been put forward in this paper to assess the performance of quinoa genotypes in new environments viz: Hypothesis (1): "Within the high genetic diversity of the species, it is possible to identify the most suitable variety for each study site" and Hypothesis (2): "To meet the various uncertainties related to global change in each region, yield stability of one variety across all study sites represents an important indication of genotype suitability and decreased risks when cultivating it any other given area." Analysis of these two hypotheses will structure the discussion on two possible pathways: yield maximization for potential production and yield optimization for production stability (Tilman et al., 2002).

### MATERIALS AND METHODS

### Study Sites

The paper concerns an analysis of the results from nine countries with similar ecological conditions. In this set, countries are located in North Africa, the Near East, and Asia under quite similar semi-arid or arid climatic conditions, namely in Algeria, Egypt, Iran, Iraq, Kyrgyzstan, Lebanon, Mauritania, Tajikistan, and Yemen.

Within each of these countries, 2–5 sites were chosen to represent the diversity of agro-ecological zones at country level. The study sites are parts of the decentralized network of agronomic stations from each of the National Research Centers involved in the programme. We tested a set of 21 different quinoa genotypes and their responses in terms of agronomic performance cultivation in these contrasted environments were duly recorded. The locations of these sites are found in **Figure 1**. **Table 1** also provides the principal descriptive data of each of 19 sites surveyed and used for this paper. The sowing dates reflect ecological and agronomic differences across study sites to adapt quinoa cycle into local cropping systems (**Table 1**).

### Genetic Resources Materials

In **Table 2**, the different genotypes used were classified in three groups considering a gradient of the level of genetic diversity among them, from landraces (heterogeneous crop population varieties) to varieties under development (still conserving a degree of heterogeneity) and improved varieties (homogeneous

plants) protected by intellectual property rights (Plant Variety Protection, PVP; Bazile et al., 2016).

Due to the difficulties to access quinoa germplasm at global level, each country made specific requests through its networks. FAO has mobilized various partnerships to collect different quinoa accessions to carry out the study. First, through a collaboration with the International Center for Biosaline Agriculture (ICBA, a non-profit International Organization) a research center based in the United Arab Emirates (UAE)— FAO could access specific varieties of quinoa. Seeds of the three varieties under development (Q1, Q2, and Q3) were obtained from ICBA to be made available for the trials in Yemen. FAO has also received seeds from the Centro di Ricerca per la Cerealicoltura (CRA-CER) in Italy which helped to expand the genotypes proposed in the tests. The seeds of nine accessions of landraces (Q12 to Q31) were obtained from CRA-CER who has been working and selecting these accessions in Italy after accessing them from the United States Department of Agriculture genebanks (USDA). These seeds had Chilean origin. Seeds were supplied to FAO-RNE which has distributed them to eight countries in the region. The two quinoa varieties (Sajama and Santamaria) were provided by PROINPA in Bolivia to the Seed and Plant Improvement Institute (SPII) in Iran. Selected seeds of early matured plants from the genotype "Sajama" have produced a new variety that was called "Iranshahr." Giza1 and Giza2 have been selected in Egypt from preliminary quinoa lines furnished by the University of Copenhagen in Denmark. Finally, Puno and Titicaca are two varieties of the Quinoa Quality Enterprise linked to the University of Copenhagen (Denmark). Regalona is the only quinoa variety with PVP developed by Von Baer Seeds for the Southern part of Chile (Von Baer et al., 2009). Finally, each country could choose from over 21 fairly differentiated genotypes.

### Experimental Protocol

Given the specificities of the FAO TCP projects, each country independently developed its experiments with similar technical support provided at regional level. Due to the difficulties of accessing quinoa genetic resources for some countries and in order to avoid biases in the analysis, a senior statistician provided advice on how to overcome such gaps (not all the countries have tested the same genotypes) and how to define and conduct solid statistical analyses in order to obtain scientifically reliable results. To be able to do that, in each test, the experimental protocol was based on a common core curriculum. For each site, the parameters used were described in the **Tables 1**, **4**.

The experimental design, a randomized complete block design (RCBD) with three replicates, represents the core protocol. The distance between two blocks was more than 1 m. Each of replicate plots is of the same area, i.e., 5 m long, 2 m wide, 10 m2 . The replicate plot had four rows of plants to define an inter-rank of 50 cm and a 25 cm distance between plants. In addition, 3–5 seeds were sown per hole of about 1 cm depth that was covered later. In few cases, irrigation was to secure germination step. Fifteen days after sowing, considering the germination rate, two plants were kept per hole by eliminating the other and considered for the experiment.

It is however important to note that the different varietal types (landraces, varieties under development, and improved varieties) were not necessarily used in each trial. On some sites,

### Statistical Analysis

of the seeds.

Estimates Yields were based on the whole plots (10 m2 ). As part of this study, two different averages were used to compare performances between genotypes and sites: the arithmetic average and the weighted average. Weighted average is important when you are dealing with frequencies or distributions. In our case, each site does not have the same number of genotypes studied, so the averages were weighted by the number of accessions used in the selected site to obtain actual weighted averages, reflecting more appropriately the results of the study for comparisons.

Considering the reality of our global experiment plan, the analysis of variance was only applied to an incomplete ANOVA table referring to some selected sites where similar genotypes were used extensively. A subset (16 sites × 11 genotypes) was extracted from the whole data set to a posteriori generate an experiment plan that is almost complete (**Table 3**).

A two-way analysis of variance was performed with the R Software to test for differences in yield across sites and genotypes. No site-genotype interaction was included in the model because of a lack of repetitions in some sites. Yield was square-rooted transformed prior to the analysis to stabilize the variance (See Supplementary Material). A one-way analysis of variance was also conducted to test for differences in square-rooted

TABLE 1 | General

characteristics

 of the study sites.

TABLE 2 | Classification of the 21 quinoa genotypes used for trials.

only landraces were tested, and other tests were only conducted with improved varieties (see **Tables 2**, **3**). These differences are explained by the difficulties to access germplasm and by the specific demands of each country. The resulting incompleteness of the experimental design has oriented our statistical analysis below. **Table 4** shows an example of a spreadsheet used to collect

and measure data in the experimentation using the international descriptor list for quinoa (Bioversity International et al., 2013). Pre-treating the seed with an insecticide and a fungicide was done before the test only, whenever such problems were found to exist in the area.

Subset (16 sites × 11 genotypes) extracted from the whole data set to a posteriori generate an experiment plan that is almost complete for the ANOVA.

yield across sites, leaving the between-genotype variability in the residual variance. The analysis was complemented with a multiple comparison test using Tuckey's method. The same analysis was conducted using genotype instead of site as the predictor.

### RESULTS

Yield was significantly different across sites and genotypes (twoway ANOVA: F-statistic for the site effect: 43.0, p < 0.001, F-statistic for the genotype effect: 11.9, p < 0.001).

Even when leaving the genotype variability into the residual error, yield was significantly different across sites (one-way ANOVA: F-statistic: 30.1, p < 0.001). Seven groups of sites out of sixteen were distinguished on the basis of yield (**Figure 2**).

Reciprocally, when leaving the site variability into the residual error, yield was significantly different across genotypes (one-way ANOVA: F-statistic: 3.3, p < 0.001). Two groups of genotypes were distinguished on the basis of yield (**Figure 3**).

## Yield Performances per Site for All the Varieties Tested

**Table 3** and **Figure 2** show that there are significant differences in yield among genotypes tested in different countries. A weighted average yield of 1.07 t.ha−1 was obtained for all varieties and all countries combined. The results demonstrate that quinoa can

be adapted in many environments while having higher yields than those that can be obtained in Bolivia or Peru (about 1 t.ha−1 ). Considering the strong differences between varieties and countries, it is then necessary to investigate more in depth the results of each of the countries involved in the project. The Tuckey test highlights a large gradient of yields across the study sites from LB1 (group g) to MA3 (group a; **Figure 2**). The results by country are described below but we can underline here that the higher yields were obtained at LB1 (g), EG1 and EG2 (f, g), and AL1 and LB3 (e, f, g). In addition, MA3 (a), MA1 and MA2 and IQ1 (a,b) and, AL2 and YE3 (a, b, c) did not performed well and yields were always lower than in other locations.

### Algeria

Two sites were analyzed and 13 genotypes were tested (**Table 3**). The yield average obtained was 1.65 t.ha−1 at the first site (AL1) and 0.26 t.ha−1 at the second site (AL2) which gives an arithmetic average of 0.96 t.ha−1 for the country. Note that the lower values at the second site, never exceeding 0.78 t.ha−1 , contributed to the fact that the arithmetic average of the country remains below the value of the weighted average yield for all countries. The three genotypes Q26 (2.62 t.ha−1 ), Q18 (2.27 t.ha−1 ), and Q27 (2.17 t.ha−1 ) achieved the much higher yields at the first site.

### Egypt

Two sites were analyzed and 13 genotypes were tested (**Table 3**). A higher value than the weighted average yield for all countries was obtained with 1.89 t.ha−1 at the first site (EG1) and 2.35 t.ha−1 at the second site (EG2). Very high yields were obtained with Q12 (3.87 t.ha−1 ), Q18 (3.17 t.ha−1 ), and Q29 (3.41 t.ha−1 ).

### Iran

Only one site has been analyzed considering the lack of observations in the other sites and 11 genotypes were tested (**Table 3**). They produced an average yield of 0.85 t.ha−1 for this site (IN3). Titicaca achieved the highest yield (4.48 t.ha−1 ) in Iran. Note however that some landraces like Q12 (1.03 t.ha−1 ) and Q21 (1.56 t.ha−1 ) have significantly higher values than the arithmetic average of the country.

### Iraq

Two sites were analyzed for this study and nine genotypes were tested (**Table 3**) for an average yield of 0.10 t.ha−1 at the first site (IQ1) and 0.65 t.ha−1 at the second site (IQ2). A value significantly lower than the weighted average yield for all countries was obtained at the two sites and an arithmetic mean for the country also much lower than all countries with only 0.36 t.ha−1 .

### Kyrgyzstan

Two sites were analyzed for the study and 3 genotypes were tested (**Table 3**) with an average yield of 0.90 t.ha−1 for the first site (KY1) and 1.03 t.ha−1 for the second site (KY2) confirming a value significantly lower than the weighted average yields for all countries at the two sites. Any improved varieties achieved yields higher than 1.24 t.ha−1 .

### Lebanon

Three sites were analyzed and 11 genotypes were tested (**Table 3**). An average yield of 2.7 t.ha−1 was obtained at the first site (LB1), 1.15 t.ha−1 at the second site (LB2) and 1.82 t.ha−1 at the third site (LB3). A value higher than the weighted average yield for all countries was obtained at the three sites. In the first site, landraces Q12 (4.0 t.ha−1 ) and Q29 (4.5 t.ha−1 ) had very high yields which gave a high amplitude to this dataset. This is also the case at the second site where Q18 (2.8 t.ha−1 ) and GIZA1 (2.8 t.ha−1 ) had a high value. This phenomenon is also present at the third site where the two cultivars (GIZA1 and GIZA2) have zero yields as did the landrace Q31. But on the other hand, Q21 (4.3 t.ha−1 ) and Q27 (3.6 t.ha−1 ) achieved very high yields. These important differences between accessions explain a significant dispersion of yields in the three sites within the dataset.

### Mauritania

Three sites were analyzed for this study and 9 genotypes were tested (**Table 3**) with an average yield of 0.08 t.ha−1 at the first site (MA1), 0.08 t.ha−1 at the second site (MA2) and 0.05 t.ha−1 at the third site (MA3). The bad arithmetic average with 0.07 t.ha−1 was explained by low yields everywhere.

#### Tajikistan

One site was analyzed and only the three improved varieties were tested (**Table 3**). The study site (TA1) achieved an average yield of 2.13 t.ha−1 . PUNO and TITICACA achieved good performance with 2.2 t.ha−1 .

#### Yemen

Three sites were analyzed and 12 genotypes were tested (**Table 3**). The yield averages obtained were 1.21 t.ha−1 at the first site (YE1), 1.14 t.ha−1 at the second site (YE2), and 0.72 t.ha−1 at the third site (YE3). Thus, one can observe a value higher than the weighted average yields for all countries at the first two sites and an arithmetic average lower than that of all countries at the third site. Q27 achieved the higher yield with 1.9 t.ha−1 at YE2, following by Q26 with 1.8 t.ha−1 at YE1 and Q1 with 1.7 t.ha−1 at YE3.

### Varieties Performances across Study Sites

**Figure 3** shows the performance for each genotype across the locations. Most of the genotypes presented higher yields than under Andean conditions in the farming systems examined. Most part of the accessions exhibited huge variations in seed yield data. Considering the large standard deviations observed for each genotype, it cannot be concluded that these accessions have a similar performance across the locations, to define the best one that can be sown with limited risks. The Tuckey test has generated two groups where Q12, Q18, Q21, and Q26 achieved the higher yields (**Figure 3**, group b) and where Q31 is represented as the lower one (**Figure 3**, group a); all the other genotypes are intermediaries.

#### Analysis by Varietal Types

#### **Landraces**

In this research project, nine landraces mainly from Chile were tested. When observing the results for each of the study sites (**Figures 3**, **4**), a first dichotomy can be made within landraces. Indeed, we find that three landraces have yields averaging lower than the overall average of 1.07 t.ha−1 : Q19 (0.89 t.ha−1 ), Q22 (1.05 t.ha−1 ), and Q31 (0.24 t.ha−1 ). The other six landraces present above-average yields: Q12 (1.40 t.ha−1 ), Q18 (1.40 t.ha−1 ), Q21 (1.38 t.ha−1 ), Q26 (1.34 t.ha−1 ), Q27 (1.24 t.ha−1 ), and Q29 (1.20 t.ha−1 ). Nevertheless, the dispersion of data of such landraces is very high and standard deviations are ranging from 1.19 to 1.35.

#### **Varieties under development**

As part of this study, eight varieties under development or without PVP (**Table 2**) were tested from different parts of the world (Bolivia, Egypt, Iran, and UAE). After reviewing the results of each of the study sites, they can be separated into two groups (**Figures 3**, **4**). Four of these varieties have average

yields below the global average of 1.07 t.ha−1 : GIZA2 (0.72 t.ha−1 ), SANTAMARIA (0.59 t.ha−1 ), SAJAMA (0.58 t.ha−1 ), and SAJAMA IRANSHAR (0.20 t.ha−1 ). The other four perform better: Q1 (1.30 t.ha−1 ), Q2 (1.13 t.ha−1 ), Q3 (1.13 t.ha−1 ), and GIZA1 (1.88 t.ha−1 ). Moreover, only GIZA1 achieved very promising performance for varieties under development.

#### **Improved varieties**

For this project, three improved varieties with PVP were also tested. These three varieties have very satisfactory performance levels (see **Figure 4**) in the few sites where there were tested. Indeed, it may be noted that these three varieties have yield averages much higher than the overall average from 1.37 t.ha−1

for REGALONA, to 1.45 t.ha−1 for PUNO and to 2.05 t.ha−1 for TITICACA, which was the best variety among all the genotypes of panel. PUNO and REGALONA varieties have high uniformity. It is not the case with the variety TITICACA, which presents significant sources of heterogeneity.

### DISCUSSION

### Germination Rates and Agronomic Practices

A very low germination rate was noted for some genotypes tested. This might be due to stocking conditions where high humidity lowers germination quality between seasons (Coulibaly et al.,

2015). Large differences in yields were observed among the tested genotypes in the different countries. If global results show that quinoa could be adapted to many environments with higher yields than in traditional cropping systems of Peru or Bolivia, we can state that it is not the case everywhere.

Different reasons could explain the differences, in particular when the seeds of some accessions showed a low degree of germination. Seed quality is the first bottleneck for experimentation because many genotypes were conserved in genebanks for a long time before being sent by plane (submitted to high pressure). Soil preparation generally constitutes the second bottleneck. Before sowing, the soil should be properly prepared to achieve a good seed-to-soil contact in the seedbed. The sowing date is another element of this second bottleneck. Preparing the soil under good weather conditions is crucial to obtain this ideal seedbed. All the conditions, including availability of seed accessions, are part of a good installation of the crop in the field avoiding too late sowing. Hot temperature is the third bottleneck and it is very important to plan the sowing date to avoid air temperatures higher than 32◦C during the flowering stage, and taking into account that harvest must be done during the dry period. Therefore, the harvested seeds of quinoa need to be well stored at a temperature of less than 15◦C under dry conditions to preserve their potential of germination.

### Access to Genetic Resources

Genebanks are mainly at the origin of development and distribution of our genotypes. However, genebanks should be distinguished as two different entities. We are interested first in genebanks from the Andean region, which can directly provide landraces from their ex situ conservation of national germplasm stocks, and secondly in genebanks outside the Andean region such as the USDA which has received seeds of that Andean area. It could also have received seeds from farmers' fields or through collaborations between genebanks. USDA collected landraces on its own account for plant breeding. In addition, USDA provides landraces from its collection to other breeders worldwide who request them in very small quantities (200 seeds) and for a limited number of accessions. While it seems easy to access this genetic material, it can be observed that the distribution is limited due to the limited access to quinoa germplasm and to the limited trade with the Andean countries because of the international regulatory context on seeds: farmers' rights, breeders' rights, phytosanitary rules, and protection of biodiversity against biopiracy. Farmers' varieties (landraces) should be recognized and protected by special agreements between research partner institutions for a particular exchange type, for example through a Standard Material Transfer Agreement (SMTA). The SMTA is a mandatory model for parties wishing to provide and receive material under the Multilateral System of the International Treaty on Plant Genetic Resources for Food and Agriculture but they are often difficult to implement. The Treaty recognizes the contribution that the local farmers of Andean regions, particularly in this case where they are located in centers of origin of crop diversity, have made for the conservation and development of plant genetic resources. Considering and using the SMTA, it gives governments the responsibility for implementing Farmers' Rights, clearly mentioned in its article 9.

Quinoa breeders can be two types: private institutes (Semillas Von Baer, Chile), public institutions (the University of Copenhagen or the CRA-CER Italy) or Non Profit International Organization such as the ICBA. If they have not developed their own collections, they could request access through genebanks, such as the USDA, in limited quantities. They multiply seeds before starting their breeding programs. After that they can benefited from results of this multiplication through the selection of landraces and assign new names. These cultivars or varieties are sold to other organizations, such as FAO that requests quinoa seeds to conduct international tests.

Following this process, there are different possible varietal types that were used in our tests: landraces, improved varieties and varieties under development. However, this genetic material could greatly change over the years and undergo several stages of selection and multiplication in different environmental conditions to those of its original environment. The characteristics of these accessions considered for our trials have probably changed significantly from their Andean nature of origin throughout all these steps.

It is important to note that this process has different pathways for seed diffusion but one can now see that they are related to each other. Being able to see them at separate stages in a continuum, we would see it more as a unique process of distribution and dissemination of genetic diversity of quinoa among different partners and institutions. The main consequence of these constraints on the access to genetic resources of quinoa is that a very low diversity is generally used to conduct trials (Bazile et al., 2016). This happens both, regarding the number of genotypes to be tested, and within the intrinsic genetic diversity of the genotypes for varieties under development and improved varieties that are developed for different environmental conditions (Murphy et al., 2016). These varieties have been selected so that they reduce their genetic heterogeneity which is synonymous to a lower adaptive capacity and contrary to high genetic diversity.

### Relationships with Original Ecotypes

In order to determine if the characteristics of the sites could be compared to those of the original ecotypes of quinoa, a comparison was made based on three criteria: altitude, latitude and annual rainfall. We noticed that in crossing the variables two by two, we come to systematically identify a group of sites that comes close to the characteristics associated with an original ecotype. In all three cases, the data is always close to the coastal quinoa ecotype that grows at sea level altitude, and latitudes corresponding to more Mediterranean climates. Nevertheless, one cannot completely involve our study sites in this ecotype because their precipitation levels are too far apart. Most study sites have annual rainfall lower than 500 mm, while in the region of origin for this coastal ecotype, from the Secano Costero in Chile's central region to the area of Araucanía in the southern Chile, rainfall ranges from 600 to 2000 mm. Landraces from this ecotype, adapted to a level of such precipitation, will not grow easily in the more arid conditions of our study cases, where rainfall rarely exceeds 500 mm, and where additionally 75% of our test sites receive less than 300 mm annually. If access to landraces was improved from the south-central area of Chile, it might be possible to make a screening of accessions to try to find this kind of highly drought resistant genotype. But as access conditions are limited, it is necessary to consider other solutions for quinoa adaptation in new environments.

No experimental site had the same ecological conditions that were observed where the original quinoa ecotypes grew. It is impossible to simultaneously align our three main experimental variables: latitude, altitude and rainfall. However, another solution is available: it is quite feasible to cross two genotypes from two ecotypes with different attributes. For example, using a variety from the ecotype of "Salares" capable of supporting arid conditions (<200 mm rainfall per year) and another variety from the "coastal" ecotype that grows in areas with low altitudes (0–1500 m). A variety capable of growing at low altitude and resistant to low levels of rainfall would be interesting for our study since we find these conditions in most of our test sites. An application of this crossing method for quinoa plant breeding was already done in the past for other attributes, to obtain the variety Regalona, a hybrid between two sources, one close to the equator and the other at southern Chilean regions (Von Baer et al., 2009).

## Discussion of the Two Strategic Objectives for Quinoa Research Related to Our Initial Hypothesis

Plant adaptation in agriculture strategies is essential for building robust production systems that are resilient to the effects of climatic change while strengthening food production for the future (Araus et al., 2008). Choosing new sowing dates or switching the cultivation to a neglected or underutilized existing crop species or variety, may prove to be a very successful move to buffer the cultivation against the negative impact of climatic change. Leveraging the potential of genetic diversity along with the deployment of improved best practices, are relatively inexpensive measures affordable by farmers, especially when accompanied by adequate extension work. On the other end, conventional plant breeding and crop's improvements are more expensive and require more technology investment for pursuing the goal of greater productivity. For example, the increase in food production in recent years has been mostly achieved considering the expansion of irrigated lands. Water use efficiency appears now as a key factor to consider from another option of view the effects of climatic change (Seckler et al., 1999). In fact, assessment of new crops and varieties diffusion of traditional and improved crop varieties needs to take more into account agronomic practices suitable and sustainable for specific ecosystems.

In addition, during the twentieth century, plant breeding for major crops has contributed to the higher increase of crop productivity for the mentioned species. But at the same time, this successful story in crop yields was limited in more recent years with a stabilization of yields for wheat and rice (Conway and Toenniessen, 1999). The situation regarding crop yields nowadays is still more complex and local context of cultivation have to be more considered for agronomic practices but also for their social and cultural dimensions (Bazile, 2015).

### Maximizing Yields

National breeding programs, which are primarily concerned on how to moderate food insecurity, may modify some aspects of the agricultural intensification derived from the Green Revolution strategy (Annicchiarico, 2002) in order to select and to produce improved germplasm, socio-economically convenient, capable of maximizing the agricultural potential of specific areas and farming systems in marginal areas, and of minimizing the occurrence of crop failures or very low yields in unfavorable years. The ultimate goal of this quinoa program is to achieve adaptation and yield stability targets, especially by using variety material with increased tolerance to prevailing biotic and abiotic stresses.

As part of our study, yield maximization strategies echoes our first hypothesis and it is important to remember: "Within the high genetic diversity of the species, it is possible to identify the most suitable variety for each study site." Based on the outcome of the studies, this first hypothesis was confirmed since there are always one or more genotypes of superior performance in one site when compared to other sites, reflecting their excellent adaptability for higher production under these particular ecological conditions. However, we should ask whether this strategy to maximize productivity, considering the performance as yield per hectare, would not be somewhat inappropriate to the context of the study in terms of food security. In addition, the tested environments are all of them located in marginal areas under extremely dry conditions and sometime saline soils that cannot permit many choices of crops that support those. This intensive agricultural model would be relevant if we sought to obtain or to increase profits in the short term, for a major sale of production surplus, for example. In our context, we are not really focusing on production intensification but rather to contribute to the food security of local populations, mostly in a subsistence farming context, that is achieving sustainable productions also in difficult areas and climate conditions. Moreover, it is important to focus on the environmental risks linked to this agricultural conventional model more based on technology intensification than on ecological intensification. As part of a high production target, we often use a significant amount of inputs, such as pesticides or chemical fertilizers, which increase the level of pollution of the local environment and present health risks to populations (Caron et al., 2009). Given the limited investment capacity of small-scale farmers in these countries, the potential yield of these varieties of quinoa will never be achieved without an initial capital contribution, in terms of expenses related to inputs, to be deducted from the final performance available for sale or supply.

### Optimization for Stability of the Food Production

From an ecological point of view, adaptation is considered as a process where the adaptedness is the level of adaptation of plant material to a given environment, and adaptability is the ability to present good adaptedness in a wide range of environments (Tigerstedt, 1994). For plant breeders, the two terms refer to a condition rather than a process. It shows the ability of plant material to be high-yielding with respect to a given environment or given conditions (Gallais, 1992). For agronomist, the accepted definitions need to consider more changes across time and space. Efforts may be developed for investigating crop genetic diversity in space and time under plant breeding programs, in relation with farmers and agronomists, for more sustainable crop production (Fu, 2015).

After demonstrating that a maximization strategy is not necessarily the most appropriate to our project, it is necessary to take a different look to find an alternative pathway for quinoa testing and introduction in new environments. We had already mentioned another strategy, raising the importance of a lasting and stable food system for the populations of the countries studied. This vision of stability, as an important dimension of food security, is embodied by our second hypothesis of work and concern directly the adaptability in a wide range of environments: "To meet the various uncertainties related to global change in each region, yield stability of one variety across all study sites represents an important indication of genotype suitability and decreased risks when cultivating it any other given area." We wanted to demonstrate, whether one or more varieties have a level of stable and efficient performance across all or most of our study sites. An average grain yield of all varieties in each

location has been used to compare the performance of each variety. All varieties were listed with a higher yield than the average yield of the site. Finally, the varieties were classified based on the number of times each variety appeared higher than the average of the different study sites. Six genotypes (Q12, Q18, Q21, Q22, Q26, and Q27) have an occurrence of up to five and present good yield potentials with great stability across sites. This result was confirmed with a good ponderation by the number of sowing sites. This gives us another perspective for less considered varieties in the trials (Q22, Q27) considering the Tuckey's method used for comparisons. The improved varieties, Puno, Titicaca (and also Regalona) performed with more than 75% but they need to be tested under more various conditions considering their good yield production, with the first and third average yield among all the genotypes.

### Strategic Choices and Their Consequences

As previously mentioned, the main objective of the FAO programme was primarily to explore ways to strengthen with quinoa cultivation the food and nutritional security of the countries involved. Our goal in fact was to achieve food security at different scales (local, national, and global). In addition, we have seen that the nutritional properties of quinoa are exceptional, especially due to its high protein levels correlated with a good balance of essential amino acids for humans. A slight amount of these grains in the daily diet would provide an interesting contribution in vegetable protein for local people which have no access to other sources of proteins.

Therefore, these two agricultural models, maximizing vs. optimization, do not tend toward the same goals and do not have the same impact on food security. They are incompatible, although they may coexist within a country in different geographical areas or for differentiated groups of farmers. As explained before, we would privilege more the food security of the countries through a yield optimization strategy that corresponds to the overall objective of the project supported by FAO for achieving adaptability and stability as requested by governments of these countries (Power, 1999). Therefore, the choice of a model that implies a specific kind of variety corresponds to the main objective. In our case, it should be preferable to choose genotypes that do not necessarily have higher yield levels. These genotypes must produce a higher average yield measured with a constant, regardless of its stability production and adaptability in marginal environments (Bazile and Weltzien, 2008; Choukr-Allah et al., 2016).

However, one should not completely ignore the model of maximizing returns (Fasoula, 2012). In fact, this strategy could be considered in those countries assuming a long term insertion of quinoa in the local agricultural landscapes and cropping systems. For example, some farmers in Morocco with larger areas are already able to take a greater risk with capital investment (equipment and inputs) to target the maximum potential of improved varieties of quinoa (Benlhabib et al., 2015).

## Conclusion: Looking for an Insertion in Local Cropping Systems

In this research, the study tested two hypotheses in order to demonstrate the adaptability of quinoa outside the Andean area. One point of discussion through different perspectives must the sustainable establishment of an alien species in a new agricultural environment. The crop introduction has to move from the stage of agronomic testing to that of assessing in practice whether quinoa can take place easily and sustainably in local farming systems around the world. Focusing on major crops cultivated in project partner countries to better know their crop cycles, we analyzed the potential inclusion of quinoa in agricultural calendars. Three groups of countries differ significantly: first when quinoa is considered a winter crop (Lebanon, Egypt, and Yemen), second when it operates as a spring crop (Kyrgyzstan and Tajikistan) and finally where it is planted as a fall crop (Algeria, Iraq, Iran, and Mauritania). Such disparities are interesting to study more in depth, considering quinoa crop introduction in a geographical area in appearance with somewhat similar climatic conditions. This consideration underlines that climate data is the main lever of action to decide the sowing date and to understand the best position in the crop calendar for water use efficiency and to avoid high temperatures at the flowering period.

The main conclusion of this study is that we were able to measure and confirm the great adaptability of quinoa in new areas with very different climatic conditions than those of the area of origin of this crop. In each of the sites studied, we were able to identify one or more genotypes with a high performance level validating the hypothesis 1 of yield quality performance. High yield potentials in single locations were shown by some genotypes like Q29 (LB1), Titicaca (IN3), Q21 (LB3), Q12 (LB1), Q29 (EG2), Q27 (LB3), Puno (TA1), Q1 (YE3). Considering yield optimization within a long term vision, some genotypes maintain a good level of stable performance regardless of the specific site conditions in which they were tested. This result allows us to also validate our second hypothesis of yield stability across sowing sites. The genotypes Q12, Q18, Q21, and Q26, averaged the best yield across 16 sites. In an agroecological perspective, quinoa biodiversity could be interesting to diversify cropping systems in order to stabilize food production.

Quinoa should be considered for diversifying cropping systems for food security and it should not be incorporated as part of an already busy period of the crop calendar cycle, and then competing with other cultures that have already demonstrated their good performances in these areas. In addition, one must not focus solely on the agricultural side of the introduction of quinoa in a local agro-ecosystem. The social component remains an important part of the adaptation of exotic species and its appropriation by the new populations. The food system of the study area may be upset with the introduction of a new food. Local populations will have to integrate quinoa in their diets and they will need to be prepared and technically and culturally accompanied to handle the new grains. There is still doubt on this point because, although quinoa has exceptional nutritional properties, development of attractive and culturally acceptable food recipes will need to be developed to reach out to both rural and urban consumers. This work will be an important endeavor to take into account when considering the introduction of quinoa cultivations outside the Andean region.

### AUTHOR CONTRIBUTIONS

MA, DB, JB, LH, IM, OM, MO, NS, AS, DS, and KM have established the multi-local trials across the different countries and have collected the data from the field. DB and CP have given a permanent technical assistance and some specific orientations for the experimental plan during the implementation of the trials and their monitoring. DB conducted the analysis on results from the multi-local trials and AV contributed to the standardization of the dataset and to the first step of statistical analysis. SP collaborated with AV and had a global overview of the writing process.

### REFERENCES

Bazile, D. (2015). Le Quinoa, Les Enjeux D'une Conquête. Versailles: Editions Quae.

### ACKNOWLEDGMENTS

The authors thank all the participants of the different TCPs for their collaboration during trials and in particular FAO Agricultural Officers from the Regional Offices and FAO Headquarters, Muhammad Dost (Regional coordinator for eight countries of FAO-RNE), Hafiz Muninjanov (Regional coordinator for KYR and TAJ), and Wilson Hugo (Technical assistance from FAO-HQ). In addition, the manuscript was edited by Diana Gutierrez Mendez and Benjamin Graub, the authors also thank them for their helpful comments. Nicolas Picard, senior statistician and modeler, has developed w us the statistical analysis. Funding was provided by FAO Technical Cooperation Programmes: TCP/KYR/3405; TCP/TAJ/3404 and TCP/RAB/3403.

### SUPPLEMENTARY MATERIAL

The Supplementary Material for this article can be found online at: http://journal.frontiersin.org/article/10.3389/fpls.2016. 00850

Fu, Y.-B. (2015). Understanding crop genetic diversity under modern plant breeding. Theor. Appl. Genet. 128, 2131–2142. doi: 10.1007/s00122-015-2585-y

**Conflict of Interest Statement:** The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

The views expressed in this paper are those of the authors and do not necessarily reflect the views or policies of FAO.

Copyright © 2016 Bazile, Pulvento, Verniau, Al-Nusairi, Ba, Breidy, Hassan, Mohammed, Mambetov, Otambekova, Sepahvand, Shams, Souici, Miri and Padulosi. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

# Quinoa for Marginal Environments: Toward Future Food and Nutritional Security in MENA and Central Asia Regions

Redouane Choukr-Allah\*, Nanduri K. Rao, Abdelaziz Hirich, Mohammad Shahid, Abdullah Alshankiti, Kristina Toderich, Shagufta Gill and Khalil Ur Rahman Butt

*Research and Innovation Division, International Center for Biosaline Agriculture, Dubai, UAE*

Quinoa is recognized as a crop of great value in terms of tolerance to abiotic stresses and there is growing interest to introduce it in marginal agricultural production systems worldwide. Also, quinoa is one of the most nutritious food crops currently known and the nutritive properties of the crop are seen as a mean to fight malnutrition globally. Various quinoa cultivars have been screened for tolerance to salinity, water-use efficiency and nutritional quality and the positive attributes found in them have created even wider global interest in its cultivation. This paper summarizes 15 years of studies on assessing the potential for introducing the crop in a few countries of the Middle East and North Africa (MENA) and Central Asia regions and describes the key constraints for scaling-up the production under marginal growing conditions in the newly introduced countries.

Edited by: *Didier Bazile, CIRAD, France*

#### Reviewed by:

*Abu Hena Mostafa Kamal, University of Texas at Arlington, USA Hector Daniel Bertero, University of Buenos Aires, Argentina*

\*Correspondence:

*Redouane Choukr-Allah<EMAIL_ADDRESS>
#### Specialty section:

*This article was submitted to Crop Science and Horticulture, a section of the journal Frontiers in Plant Science*

Received: *31 December 2015* Accepted: *07 March 2016* Published: *29 March 2016*

#### Citation:

*Choukr-Allah R, Rao NK, Hirich A, Shahid M, Alshankiti A, Toderich K, Gill S and Butt KUR (2016) Quinoa for Marginal Environments: Toward Future Food and Nutritional Security in MENA and Central Asia Regions. Front. Plant Sci. 7:346. doi: 10.3389/fpls.2016.00346*

Keywords: irrigation, salinity, yield, sowing dates, food security

### INTRODUCTION

Quinoa has been recognized as a climate resilient crop of great value and there is an increasing effort to introduce it in different marginal agriculture production systems of the world. Various quinoa cultivars have been screened for tolerance to abiotic stresses, especially salinity, drought, and frost and the positive attributes of the crop have created wider global interest in its cultivation (Jacobsen, 2003; Jacobsen et al., 2003). Also, quinoa is one of the most nutritious food crops currently known. The seeds contains high quality protein, which has all of the essential amino acids including lysine, methionine and threonine that are scarce in cereals and legumes (Repo-Carrasco et al., 2003). In view of its exceptional nutritional quality and ability to grow under marginal environments, the Food and Agriculture Organization of the United Nations (FAO) has identified quinoa as one of the crops that will play an important role in ensuring future food security and designated the year 2013 as the "Year of Quinoa" (Bazile et al., 2015). Worldwide, the demand for quinoa is growing, especially in the health food segment, but current supplies are unable to match it. Besides the use for human consumption, quinoa seed has other uses as livestock and poultry feed. The whole plant can be used as green fodder and harvest residues can be fed to the animals. In the context of the Middle East and North Africa (MENA) and Central Asia regions, where soil and water salinity is increasingly becoming a constraint to agricultural production, quinoa is seen as an alternative crop with significant potential to have a central role in sustaining farm productivity.

The International Center for Biosaline Agriculture (ICBA) in Dubai, United Arab Emirates (UAE) has been working since 2007 on quinoa in partnership with the Ministry of Environment and Water (MOEW) of the UAE, Abu Dhabi Farmer's Service Center (ADFSC) and its Peruvian partners [Instituto Nacional de Innovacion Agraria (INIA) and Universidad National Agraria La Molina (UNALM)] to evaluate the performance of several quinoa cultivars for their productivity when grown in marginal conditions. ICBA has identified and developed five high yielding salt- and heat-tolerant lines that are now ready to be tested in other agro-ecological zones. The FAO has also been in the forefront of quinoa research, keeping in view its unique nutritional characteristics and adaptability to a wide range of agro-ecological conditions, especially in the marginal areas and the organization has initiated a Regional Project TCP/RAB/3403 for the introduction, appropriation and institutionalization of its production in Algeria, Egypt, Iran, Iraq, Lebanon, Mauritania, Sudan, and Yemen. ICBA considers quinoa as an essential crop in its efforts to sustain agricultural productivity amidst growing threat of salinity and water scarcity, as well as to tackle hunger, malnutrition and poverty in the MENA and Central Asia regions. To fully exploit the potential of the crop for marginal environments, identification of new and high-yielding quinoa genotypes with good local adaptation and high nutritional quality are crucial, which requires intensified screening and adaption research.

This paper summarizes 15 years of studies in assessing the potential of quinoa for introducing it in some countries of the MENA and Central Asia Regions. In view of quinoa's potential as a stress-tolerant, climate resilient future-proof crop, ICBA is developing best practices for its production and management in marginal environments. The Center is also focusing on utilization and marketing of the produce and on fostering knowledge transfer, strengthening competencies and exchange of experiences among the various stakeholders within the MENA and Central Asia regions.

### SCREENING FOR LOCAL ADAPTATION AND YIELD POTENTIAL

ICBA acquired 121 germplasm accessions from the United States Department of Agriculture (USDA) and evaluated them for growth performance and yield at its research station (25◦ 05′ 49′′ N and 55◦ 23′ 25′′E) during the cropping season November 2006– March 2007. The soil at the experimental site was fine sand and moderately alkaline (pH 8.2) with very low organic matter (<0.5%). The seeds were sown in the first week of November 2006 and each accession was planted in three 3 m-rows with spacing of 50 cm between the rows and one meter between two accessions. The distance between plants within each row was maintained at 25 cm. Before sowing, the fertility of the soil was improved by incorporating organic fertilizer (compost) at the rate of 40 t ha−1 and during crop growth two split doses of NPK (20:20:20) at the rate of 50 kg ha−1 , were applied by banding alongside the rows. The amount of N added to the soil by the combination of compost and NPK was estimated to be 570 kg ha−1 . The plants were irrigated with low-salinity water with an electrical conductivity (ECw) of 2–3 dS m−1 using the drip system. Water was applied once every day for 20 min at a flow rate of 4 l h−1

per plant and in addition, about 60 mm of rainfall was received during the growing season. Of the 121 accessions planted, seeds were harvested from 73 accessions that survived through the growing season and based on the seed yield data, the top 20 performers were selected for subsequent evaluation, carried out during the cropping seasons 2007-08 and 2008-09 (Rao and Shahid, 2012). The agronomic practices such as irrigation and applications of fertilizers were the same as described previously. The data on grain yield of the 20 accessions from the two cropping seasons (2007-08 and 2008-09) are presented in **Table 1**. The seed yield among the accessions varied between 53.86 and 359.86 g m−2 in 2007-08, and between 3.32 and 258.42 g m−2 in 2008-09. Averaged over the two seasons, the yield ranged between 34.06 g m−2 and 238.99 g m−2 among the accessions. The studies showed that quinoa has good adaptation and can be successfully cultivated in the Arabian Peninsula. In a further study conducted at the ICBA Research Station in Dubai during 2009-10, the performance of five top-yielding accessions selected on the basis of average seed yield from previous studies were further evaluated using low-salinity irrigation water (ECw 2.8 dS m−1 ) (Rao and Shahid, 2012). Analysis of the data revealed significant differences for plant height, number of primary branches and number of inflorescences per plant. However, differences among accessions for all other traits including inflorescence length, seed yield per plant, fresh and dry biomass were found to be marginal. The seed yield among the five accessions ranged between 374.4 g m−2 (Ames 13757) and 533.6 g m−2 (Ames 13761) with an average of 456.6 g m−2 over accessions. The dry matter yield, averaged over the accessions was 1464 g m−2 , with accession Ames 13742 producing the maximum yield of 1624 g m−2 (Rao and Shahid, 2012). During course of the yield trials over the years, mass selection to eliminate inferior plant types and improve the yield potential of the selected accessions has resulted in five improved lines (Q1–Q5) which were used in further studies described below.

### AGRONOMIC EVALUATION AND PERFORMANCE OF SELECTED LINES

The results from the preliminary trials undertaken at ICBA Research Station gave sufficient indication of the potential of quinoa as a salt-tolerant alternative crop. However, further investigations were needed to study the performance under a range of biophysical environments especially at various soil and water salinities to introduce the crop to the farmers. Hence field trials were conducted under a range of agro-ecological conditions—at Ghayathi in the Western Region of Abu Dhabi Emirate in 2012-13 with two of the five selected lines (Q3, Q5) and NSL 106399; and subsequently in five locations—three in the Northern Emirates (Dibba, Hamraniah, and Al Dhaid) and two in the Western Region of Abu Dhabi (Madinat Zayed and Ghayathi) in 2013-14, involving four of the five selected quinoa lines (Q1, Q2, Q3, and Q5) (**Table 2**). While the two locations planted in the Western Region were agricultural farms leased for the trials, those in the Northern Emirates were the federal government's regional research stations. Each set of lines was

TABLE 1 | Grain yields in 20 selected quinoa accessions grown in winter 2007-08 and 2008-09 (source: Rao and Shahid, 2012).

sown in a randomized complete block design (RCBD) with three replications. In all cases, the distance between rows was 50 cm and between plants within the row was 25 cm. Spacing between two adjacent plots was maintained at 1 m. The amount of water and fertilizers applied were the same for all the five locations and as described in the previous section.

In 2012-13, at Ghayathi, despite high salinity of irrigation water (ECw14-15 dS m−1 ), the mean seed yield obtained of the three lines was 750 g m−2 which was on par with the highest yields reported from the non-saline traditional quinoa growing areas. The green biomass yield was also high, the mean of the three cultivars being 4.3 kg m−2 , indicating the potential of quinoa as an alternative forage crop for saline areas (Rao et al., 2013). In 2013-14 trials, the salinity of irrigation water in the five locations varied between 2.3 and 18.9 dS m−1 (**Table 2**). Seed yield from trials in the Northern Emirates correlated with the salinity of irrigation water (ECw), which was 6.1 dS m−1 in Dibba, 4.5 dS m−1 in Hamraniah and 2.3 dS m−1 in Al Dhaid. Thus, averaged over lines, the seed yield was highest in Al Dhaid (541 g m2 ), followed by Hamraniah (398 g m−2 ) and Dibba (190 g m2 ) (Rao, 2016; **Figure 1**). In the western region despite higher salinity of the irrigation water, significantly higher seed yields were recorded than those from the Northern Emirates. In Ghayathi, the mean seed yields of the four lines was 1050 g m−2 , much higher than the yields recorded in the previous year (750 g m−2 ) from the same location. In Madinat Zayed the mean seed yield

TABLE 2 | Quinoa yield trails in the UAE: Soil and water characteristics at the locations and details of accessions evaluated (Rao, 2016).

of the four lines was 700 g m−2 , which was similar to the yield obtained from Ghayathi in the previous year (Rao et al., 2013; Rao, 2016; **Figure 1**). The results from yield trials showed high degree of variability in the performance of the different cultivars across locations (**Supplementary Figure 1**). While no major differences existed among the different locations in terms of radiation, mean maximum and minimum temperatures or precipitation during the growing period, the differences in yields could only be attributed to differences in physicochemical properties of the soils and water (other than the EC and pH which were measured), for which more detailed analyses are warranted. The two locations planted in the Western Region being agricultural soils, possibly had a more favorable microenvironment compared to the sandy soils with poorly developed profiles at the research stations in Northern Emirates. Nevertheless, the exceptionally higher yields obtained with highly saline irrigation water in the Western Region showed that quinoa, which is a facultative halophyte, not only endures salinity but also some of its cultivars prosper under saline conditions, therefore has huge potential as an alternative food and feed crop when growing traditional crops becomes uneconomical due to increased groundwater salinity. The results besides confirming quinoa's suitability to withstand high salinity in water and soils, also demonstrated that it has good adaptation to the hyperarid desert environments and is an excellent candidate for crop diversification in the UAE as well as other countries with similar climatic conditions.

In addition to the UAE, ICBA has recently undertook pilot trials in six other MENA and Central Asian countries and the selected quinoa lines proved to be very productive even under very poor soils (sandy soils) and under conditions of deficit irrigation and high salinity. For instance, in 2015, five quinoa lines (Q1–Q5) were evaluated for seed and forage yields in three countries of Central Asia, namely: Uzbekistan, Tajikistan and Kyrgyzstan under different eco-agroclimatic zones that significantly differ in soil characteristics and agricultural practices (ICBA, 2015). Results from Kyrgyzstan were variable due to late planning but in Uzbekistan the seed yields among the lines ranged between 294 g m−2 (Q3) and 557 g m−2 (Q5) with a mean

of 436 g m−2 and in Tajikistan they ranged from 147 g m−2 (Q2) to 336 g m−2 (Q1) with a mean of 227 g m−2 (**Figure 2**).

The high yields obtained under marginal agro-climatic conditions in different locations are indicative of the potential and further investigations are needed to study the performance of a much wider range of genetically diverse accessions at various soil and water salinities to fully exploit the available genetic diversity within the crop. Quinoa is a predominantly self-pollinating species and considerable variation exists between cultivars for many of the desired characters. Therefore, it should be possible to select better adapted genotypes with high yields and nutritional quality combined with salt-and drought-tolerance. Identification of desirable genotypes needs to be followed by work on optimization of cultural practices to maximize productivity under the local conditions.

Quinoa was evaluated and introduced in Morocco for the first time in 2000-01 growing season in two locations, Khenifra (mountain region) and Rabat (coastal region). Obtained results are presented in **Figure 3**. Among the tested lines, G205-95DK performed the best under high altitude conditions due to its origin in the high altitudes of Altiplano (Andean Plateau), while most of the other lines showed adaptation to the coastal region probably because of provenance effect (Benlhabib et al., 2015).

In Morocco, in a field experiment conducted in Rhamna region, four selected quinoa lines (L11, L119, L123, L142, L143) along with two commercial varieties Titicaca and Puno were evaluated for phenological and agro-morphological characteristics (Filali, 2011). Titicaca and L143 recorded the highest seed yield (1.5 t ha −1 ), while L11 showed the lowest seed yield (0.46 t ha−1 ). The same lines when cultivated in the experimental farm of the Agronomic and Veterinary Medicine Hassan II Institute, Horticultural complex, Agadir (IAV-CHA) and tested under four different irrigation levels (100, 75, 50, and 25% of full irrigation), responded differently to water stress (**Figure 4**). The line L143 was most affected by water stress, L123 and L11 were less affected as the reduction in yield compared to the fully irrigated treatment was less (**Supplementary Figures 5**, **6**).

### NUTRITIONAL QUALITY

Quinoa seeds are an exceptionally nutritious food source, owing to their high protein content with all essential amino acids, lack of gluten, and high content of several minerals such as Ca, Mg, Fe, and it is also rich in vitamins. Abiotic stress is known to induce considerable changes in the composition and quality of cereal grains such as wheat (Ashraf, 2014). Despite quinoa's growing popularity, the knowledge about the relationship between the growth conditions and the nutritional profile is still limited. Characterization of the nutritional and anti-nutritional properties of quinoa and evaluation of the effects of extreme growing conditions on the nutritional quality will provide strategic information for the introduction and promotion of quinoa in new environments besides aiding in the selection of nutritionally high and stable cultivars. Results from analysis of the chemical composition of the seeds harvested from three locations (Al Dahid, Ghayathi and Madinat Zayed) with varying levels of irrigation water salinity (2.3, 16.3, and 18.9 dS m−1 , respectively) showed that salt stress has marginal effect on the proximate composition (Rao, 2016). However, significant differences were found in the mineral content of the seeds. Especially, seeds harvested from Ghayathi and Madina zayed had higher Na content, but lower levels of Ca and Fe compared to those harvested from Al Dhaid (Rao, 2016). These findings have significant implications, especially for introduction of quinoa in highly saline areas, therefore detailed investigations are recommended for confirmation.

### AGRONOMIC TRIALS AND CULTIVATION

In the FAO's Regional Project TCP/RAB/3403, 11 quinoa cultivars were evaluated for seed yield potential in seven of

FIGURE 2 | The mean seed yields of five quinoa lines grown at four locations in Central Asia. The error bars represent standard errors of the means (ICBA, 2015).

the eight participating countries (Egypt, Iraq, Iran, Lebanon, Mauritania, Sudan and Yemen) during January-June 2014. Large yield variations per hectare were observed across locations within and between countries, the mean seed yields ranging from 0.11–0.96 t ha−1 in Iraq, 0.24–1.90 t ha−1 in Yemen, 0.41–3.87 t ha−1 in Egypt, 1.50–7.50 t ha−1 in Lebanon, 0.16– 1.56 t ha−1 in Iran, and 0.03–0.23 t ha−1 in Mauritania. The highest yield (7.50 t ha−1 ) was recorded in Lebanon, followed by Egypt (3.87 t ha−1 ), while the lowest yield was recorded in Mauritania (0.23 t ha−1 ) (Dost, 2015). A comparison of the cultivars across the countries/locations revealed very interesting and consistent patterns of performance that resulted in the identification of several potential/suitable varieties for the countries. The cultivars Q12 and Q21 exhibited outstanding performance in almost all the countries of evaluation, while Q29, Q18, Q19, Q22, Q26, and Q27 produced superior yields as compared to other cultivars at all locations/countries except Iran. Convinced by quinoa's performance, one private agriculture company in Egypt and a farmer in Lebanon planted quinoa on a large-scale (1– 2 hectares) with a plan to further extend the production together with improvement in capacity of farmers and generating market demand. However, private sector involvement in quinoa production will mainly depend on the conducive policies of the governments, allocation of financial and human resources for research and development of new varieties along with the improvement in the production technologies, processing and utilization within the region.

## CROPPING PRACTICES OF QUINOA PRODUCTION

### Deficit Irrigation

In quinoa, deficit irrigation (DI) strategy has been widely investigated as a valuable and sustainable production strategy in regions where intra-seasonal dry spells are of considerable importance (Garcia et al., 2003; Geerts et al., 2008). By limiting water applications to drought sensitive growth stages, the practice aims to maximize water productivity and to stabilize, rather than maximize yields (Geerts and Raes, 2009). Several experiments were conducted at IAV-CHA during two growing seasons 2010 and 2011 in order to evaluate the effect of deficit irrigation with treated wastewater (Hirich et al., 2012a,b,c, 2013). During the first season (2010), the response of the line D0708 with six deficit irrigation treatments (during vegetative growth, flowering, grain filling, during both vegetative growth and flowering stage, alternating with 100% of full irrigation as non-stress condition, and 50% of full irrigation as water deficit condition) were studied. During the second season (2011), two lines (D0708 and QM1113) were studied with six other irrigation treatments (rainfed, 0, 25, 50, 75, and 100% of full irrigation) only during the vegetative growth stage, while in the remaining stages, full irrigation was provided except for the rainfed treatment. The results clearly showed that the highest yields were obtained in the fully irrigated treatment followed by the treatment stressed during vegetative growth stage. When subjected to deficit irrigation during flowering, grain filling, during both vegetative growth and flowering, and during all crop stages, a yield reduction of 32, 37, 46, and 50% was recorded respectively, in comparison with the control (full irrigation). The finding indicates that applying deficit irrigation during vegetative growth stage results in increased root to shoot ratio. Early water deficit induced root development and full irrigation applied during flowering and grain filling allowed the plants to produce more biomass due to the well-developed root system, possibly leading to higher levels of water and nutrient uptake. The increased yield observed in the treatment stressed during vegetative growth stage is probably due to maximized crop water productivity (CWP), which further allowed up to 19% water saving compared to full irrigation.

Statistical analysis carried out on grain yield of quinoa in response to several degrees of water stress applied during the vegetative growth stage revealed highly significant differences among the treatments (p < 0.001). The results clearly showed that line DO708 showed the highest grain yield compared to QM1113. While applying 50% of full irrigation during the vegetative growth stage resulted in 10 and 8% increase in seed yield in DO708 and QM1113 respectively compared to control (fully irrigated), applying 25% of full irrigation caused a yield reduction of 31 and 38% (in DO708 and QM1113 respectively, compared to the control). It is obvious that CWP is maximized under 50% deficit irrigation during the vegetative growth stage in both the lines.

Results from field trials conducted in sandy soil during two growing seasons (2011 and 2012) at the Experimental Station of Cadi Ayyad University located in 70 km south West Marrakech (Morocco) and also under rainfed condition in a farmer's field at Tnin Bouchan, about 250 km away from Agadir indicated that deficit irrigation (50 and 33% of full irrigation) affected quinoa performance and resulted in seed yield reduction of 15.8, 30.1% respectively in the first season (2011) and by 15.2, 41.5% respectively, in the second season (2012) in comparison with full irrigation. Under rainfed conditions, seed yields were reduced by 62.1 and 59.3% in the 2011 and 2012 growing seasons, respectively compared to full irrigation. CWP was maximized in the treatment receiving 50% of full irrigation (Fghire et al., 2013).

### Organic Amendment

One of the most important factors which limits crop growth and production is water scarcity. As already discussed, studies show that deficit irrigation is a judicious strategy to increase water use efficiency and CWP in water-scarce environments. Organic Choukr-Allah et al. Quinoa for Marginal Environments

amendments, on the other hand, improve the water holding capacity of soils and combing them with deficit irrigation can be a practical solution for sustainable yields as the negative effect of water scarcity can be compensated through the increased water holding capacity of the soils and there by improved availability of water and nutrients to the plants. In line with this, a study was conducted in at IAV-CHA to evaluate the combined effect of organic amendment (compost) and deficit irrigation using treated wastewater on productivity of the quinoa line (D0708) during October 2011 to March 2012 (Hirich et al., 2013, 2014a). Three levels of organic amendments (0, 5, and 10 t ha−1 ) combined with two levels of deficit irrigation (50 and 100% of full irrigation) were studied. While the highest seed yield (66.3 g plant−1 ) was recorded when quinoa was subjected to full irrigation and received 10 t ha−1 of compost, yield was low under water deficit conditions without organic amendment (**Figure 5**). The results indicated that organic amendment of 10 and 5 t ha−1 increased grain yield by 16 and 3% respectively, under full irrigation conditions, and by 18 and 13% under deficit irrigation (respectively). It can be concluded that organic amendment improve quinoa yields significantly under deficit irrigation conditions by improving the water holding capacity of the soils and the access to water and nutrients by the plants. Combining deficit irrigation and organic amendment also allowed maximizing the CWP.

### Quinoa Fertilization

The combined effect of water stress and nitrogen application was studied on a quinoa line (D0708) in a field trial laid out in split-plot design at IAV-CHA, during the year 2013 (**Supplementary Figures 2**, **4**). The treatments included four levels of water stress (25, 50, 75, and 100% of full irrigation) and seven different rates of nitrogen (0, 40, 80, 120, 160, 200, and 240 kg ha−1 ) application. The results suggested that seed yield increased with increasing nitrogen supply, though the response varied with the level of water stress (**Figure 6**). The yield was highest in the 50% of full irrigation treatment with 240 kg ha−1 of nitrogen. CWP increased with higher supply of nitrogen and the degree of water stress, the value being highest in the most stressed treatment (25% of full irrigation) and 240 kg ha−1 of nitrogen and lowest with the full irrigation without nitrogen supply (Hirich, 2014).

In a field trial conducted in Wadi El-Natroon region, Beheira Governorate, Egypt during 2008/2009 and 2009/2010 winter seasons, Shams (2011) studied different rates of nitrogen fertilization (0, 90, 180, 270, and 360 Kg N ha−1 ) for improvement of growth and yield in sandy soils. High nitrogen fertilizer rate significantly increased yield during both the seasons. However, nitrogen use efficiency has reduced with increased rate of nitrogen application. In a pot experiment with two quinoa lines (Quinoa-52 and Quinoa-37) and two commercial varieties (Titicaca and Puno). Lavini et al. (2014) studied the effect of five rates of nitrogen application (0, 50, 100, 150, and 200 mg kg−1 of soil). The results showed that both the lines responded similarly to the application of nitrogen and yield has improved significantly with increased nitrogen rate.

In studies conducted by Gomaa (2013) in Egypt, quinoa plants were fertilized with ammonium nitrate (34% N) at 0, 120, 238, 357 kg ha−1 in combination with nitrobin (biofertilizer) or calcium super phosphate (15.5% P2O5) at 0, 120, 238, 357 kg ha−1 in combination of phosphorin (biofertilizer). The plants performed the best in the treatment receiving 238 kg ha−1 of ammonium nitrate in combination with nitrobin.

### Use of Saline Water

A pot experiment was carried out at the IAV-CHA to evaluate the response of quinoa (line D0708) to different levels of irrigation water salinity (1, 10, 20, and 30 dS m−1 ) (Hirich et al., 2014c,d; Lavini et al., 2014). Irrigation with water at 10 dS m−1 salinity has not affected seed yield significantly and yield reduction was only 9% compared to the fresh water control (1 dS m−1 ). Increasing salinity to 20 dS m−1 and then to 30 dS m−1 resulted in a significant reduction in seed yield (24 and 34%, respectively, compared to the control. Contrarily, saline water irrigation had positive effect on CWP—the most efficient treatment being the most salt-stressed treatment (30 dS m−1 ), which produced 0.8 kg for 1 m3 of water and the lowest being the control (0.6 kg m−1 ) (p < 0.001). This difference in terms of CWP can be explained in terms of the difference in water uptake which was higher in the fresh water treatment and lower in the saline treatments.

Three lines of quinoa (QM1113, QS0938, D0708) cultivated in open field were irrigated with treated wastewater with different levels of salinity in order to assess the impact on yield (El Youssfi et al., 2012). Significant differences were found in the performance of the lines—QM1113 being the most productive with an average grain yield of 6.92 t ha−1 at 6 dS m−1 salinity followed by D0708 with an average yield of 5.65 t ha−1 at 3 dS m−1 . The findings indicated that quinoa yields increase with increasing salinity of up to 6 dS m−1 and being a facultative halophyte, salinity promotes growth but up to a certain threshold, beyond which growth and productivity start to be negatively affected. Results from these studies differ with those of Rao (2016) from on-farm trials where seed yields were maximum despite irrigation with highly saline water (15–18 dS m−1 ); and those from field experiments in Turkey where salinity stress of up to 40 dS m−1 did not interfere with seed and biomass yields significantly (Yazar et al., 2015); or of Razzaghi et al. (2011) who showed an yield reduction of 50% with saline water of 40 dS m−1 compared with fresh water.

Simple seed germination tests were conducted at ICBA, Dubai to study the role of BonteraTM (Biofertilizer and humic acid) in enhancing seed germination and in mitigating the negative impact of salinity (**Figure 7**). Different levels of sodium chloride of up to 50% seawater salinity (1.2% NaCl) were tested in combination with different dilutions of BonteraTM (ranging from 1:20 to 1:500). The results of these tests showed that salinity in the growth medium inhibited seed germination to the extent that at 1.6% NaCl seed germination did not occur. At lower concentrations of BonteraTM (dilutions of 1:500 and above) significantly enhanced seed germination and seedling growth, while higher concentrations (1:300) were inhibitory (Gill et al., 2016). Under field conditions, foliar application of BonteraTM consistently showed up to 28% improvement in grain yield. It was concluded that application of BonteraTM at 1–2 l ha−1 could reduce the chemical fertilizers application by 25%. Foliar application of BonteraTM in combination with two irrigation

FIGURE 5 | Combined effect of organic amendment and deficit irrigation on quinoa yield and water productivity (Hirich et al., 2014a).

regimes (100% ET and 50% ET) showed greater impact at 50% ET suggesting a mitigating effect for stresses like water deficiency and drought (Gill et al., 2016).

### Sowing Dates

Hirich et al. (2014b) carried out a series of field trials at IAV-CHA (**Supplementary Figure 3**), to investigate the effects of 10 sowing dates—from 1st November to 15th March with 15-day intervals, on quinoa performance. The study showed that sowing dates affected growth and productivity obviously due to differences in temperature, precipitation and radiation over time (**Figure 8**). Seed and dry matter yields were highest when quinoa was sown in November and early December. Reduced yield during the late sowing dates was explained by delay in germination due to prevailing low temperatures and occurrence of downy mildew during February and March triggered by high air humidity. The length of growing period increased from November to January and decreased from January to March. The longest growing period was when sown on 1st of January and the shortest was when sown on 15th of March.

 In the trials conducted in the UAE and the FAO project countries, significantly high variability was observed in the yields of the lines/varieties. Therefore, understanding of the influence of edaphic and climatic factors on productivity in different locations is important. There

is also considerable knowledge gap regarding quinoa pests and diseases, particularly in areas outside its traditional growing regions. Some of the wild relatives such as Chonopodium murale and C. album are common weeds in several of the countries where quinoa is introduced and they possibly harbor a number of biotic stressors that can easily migrate onto cultivated species. Introduction and scaling-up of novel crops such as quinoa to non-traditional environments also requires the study and establishment of the entire chain and building capacity of the researchers and the farmers in the basics of production and harvesting, storage and processing technologies (Rao, 2016).

### CONCLUSIONS AND RECOMMENDATIONS

In recent years, quinoa is receiving significant attention as a nutritively rich multi-purpose agro-industrial crop that can thrive in extreme soil and climatic conditions. Faced with the challenge of increasing the production of high-quality food to feed the growing population, quinoa offers an excellent alternative to ensure food and nutrition security in marginal environments in the MENA as well as other regions across the globe. In the MENA region, while many countries have recently initiated work on quinoa, Morocco and the UAE have made significant advances toward introducing the crop in the local production systems. In Morocco, IAV HASSAN II Institute has been studying quinoa since 2000 as a drought and salinitytolerant alternative crop to contribute to the food and nutrition security of people in the mountainous areas. As part of this program, quinoa has already been introduced to a dozen sites across the country and each year, the number of quinoa growers' is increasing all over the regions, especially Oujda, Benslimane, Fes, Boulmane, and Marrakech. However, quinoa yields over the years remained unpredictable and very low, averaging between 1.2 and 1.4 t ha−1 , while the maximum attainable yield can be up to 8–10 t ha−1 . A range of factors such as the choice of cultivars, optimal sowing date and nutrient management were suggested to affect the production. Constraints which still need to be overcome include crop stand establishment, sensitivity to high temperatures and salinity, weed control and saponin removal. Furthermore, there is also a need to design a product marketing strategy and raise awareness among farmers and the government agencies about quinoa's potential as a stress-tolerant alternative crop for marginal environments.

In the UAE, ICBA has been investigating quinoa since 2006 as an alternative crop for salt-affected areas. Significant progress has been made in identifying salt-tolerant lines combined with high yield potential. These lines are now being further evaluated for their yield potential in several countries including Uzbekistan, Tajikistan and Kyrgyzstan in Central Asia (through the project "Cross-regional partnerships for improving food and nutritional security in marginal environments of Central Asia" funded by the Islamic Development Bank); and Yemen, Jordan, and Egypt in the MENA region [as part of the collaborative project "Adaptation to climate change in West Asia and North Africa marginal environments through sustainable crop and livestock diversification" funded by the International Fund for Agricultural Development (IFAD), OPEC Fund for International Development (OFID)]. As with any other new crop, one of the key factors for successful introduction and establishment of quinoa under the novel climates will be the identification of appropriate planting material. It is therefore important to study the adaptation and yield potential of several quinoa genotypes from different provenances to select the most promising genotypes suitable for the local agroclimatic conditions. Information on these aspects as well as the economic assessment of the profitability of quinoa cultivation is essential, especially when planted under sub-optimal growth conditions with low-quality water in the targeted countries. ICBA's future research on quinoa focuses on developing best practices in relation to the production, management under diverse farming systems and agro-ecological regions, and on utilization and marketing of the produce, besides fostering knowledge transfer, strengthening competencies and exchange of experiences among quinoa researchers within the MENA and Central Asia regions.

It is well known that climate change is rapidly degrading the conditions of crop production. Salinization and aridity are forecasted to increase in most parts of the world, especially in the MENA region. As a consequence, new stress-tolerant genotypes of the existing crops or new and alternative crops or species must be identified and used for future food security. Stresstolerant crops such as quinoa offer major opportunities due to their comparative advantage over the staple food crops in terms of tolerance to harsh growing conditions. The results presented in this paper have indeed shown that quinoa maintains productivity in rather poor soils and under water stress conditions and high salinity. Moreover, quinoa seeds are an exceptionally nutritious food source, owing to their high protein content with all essential amino acids, lack of gluten, high mineral content (e.g., Ca, Mg, Fe), and health-promoting compounds such as flavonoids. Higher yield do not guarantee quinoa's success in the region and to be successful it must fit in the current cropping patterns, farming systems and prove its worth in rigorous and properly designed on-farm trials in marginal lands affected by salinity and alkalinity and in the areas where the majority of the food crops could not be produced economically. For scaling up and large scale adoption by the farmers in the region, creating opportunities for marketing the produce are also vital. Quinoa being a new crop to the region, more efforts will be needed to create awareness for its incorporation in the social, cultural and dietary habits to increase the market demand besides strengthening the efforts to improve yields combined with nutritional quality, production, harvesting and post-harvesting practices along the value chain.

### AUTHOR CONTRIBUTIONS

RC was the corresponding author collecting data and results as well as editing results related to North Africa region. NR was providing research results conducted by ICBA. He was reviewing the paper and editing the English writing. AH contributed to this paper by designing the figures, commenting the data and compiling all gathered results. He was also revising the formatting according to Journal Author guideline. KT provided results of research undertaken in Central Asia. Butt has been involved with Dr. NR in several experiment on quinoa. SG provided results related to seed treatment in order to improve crop tolerance to salinity. AA provided results related to seed treatment in order to improve crop tolerance to salinity.

### SUPPLEMENTARY MATERIAL

The Supplementary Material for this article can be found online at: http://journal.frontiersin.org/article/10.3389/fpls.2016. 00346

Supplementary Figure 1 | Quinoa panicle variability in terms of color and panicle volume.

Supplementary Figure 2 | Trial on combined impacts of water stress and nitrogen application carried out in Agadir, Morocco.

Supplementary Figure 3 | Sowing date trial carried out in Agadir, Morocco.

Supplementary Figure 4 | Quinoa harvest within field trial on combined impacts of water stress and nitrogen application carried out in Agadir, Morocco.

### REFERENCES

#### Supplementary Figure 5 | Field trial about responses of seven quinoa accessions to several irrigation levels.

Supplementary Figure 6 | Genetic and morphological variability of several quinoa accessions.

**Conflict of Interest Statement:** The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Copyright © 2016 Choukr-Allah, Rao, Hirich, Shahid, Alshankiti, Toderich, Gill and Butt. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

# Elevated Genetic Diversity in an F2:6 Population of Quinoa (Chenopodium quinoa) Developed through an Inter-ecotype Cross

Ouafae Benlhabib1 , Noura Boujartani1 , Peter J. Maughan2 , Sven E. Jacobsen3 and Eric N. Jellen2 \*

1 Departement de Production, Protection et Biotechnologies Vegetales, Agronomic and Veterinary Institute Hassan II, Rabat, Morocco, 2 Department of Plant and Wildlife Sciences, Brigham Young University, Provo, UT, USA, 3 Department of Plant and Environmental Sciences, Faculty of Life Sciences, University of Copenhagen, Copenhagen, Denmark

#### Edited by:

Antonio M. De Ron, Misión Biológica de Galicia-CSIC, Spain

#### Reviewed by:

Pedro Revilla, Misión Biológica de Galicia-CSIC, Spain Svetla Sofkova-Bobcheva, Maritsa Vegetable Crops Research Institute, Bulgaria

> \*Correspondence: Eric N. Jellen<EMAIL_ADDRESS>
#### Specialty section:

This article was submitted to Crop Science and Horticulture, a section of the journal Frontiers in Plant Science

Received: 30 May 2016 Accepted: 02 August 2016 Published: 17 August 2016

#### Citation:

Benlhabib O, Boujartani N, Maughan PJ, Jacobsen SE and Jellen EN (2016) Elevated Genetic Diversity in an F2:6 Population of Quinoa (Chenopodium quinoa) Developed through an Inter-ecotype Cross. Front. Plant Sci. 7:1222. doi: 10.3389/fpls.2016.01222 Quinoa (Chenopodium quinoa) is a seed crop of the Andean highlands and Araucanian coastal regions of South America that has recently expanded in use and production beyond its native range. This is largely due to its superb nutritional value, consisting of protein that is rich in essential amino acids along with vitamins and minerals. Quinoa also presents a remarkable degree of tolerance to saline conditions, drought, and frost. The present study involved 72 F2:6 recombinant-inbred lines and parents developed through hybridization between highland (0654) and coastal (NL-6) germplasm groups. The purpose was to characterize the quinoa germplasm developed, to assess the discriminating potential of 21 agro-morpho-phenological traits, and to evaluate the extent of genetic variability recovered through selfing. A vast amount of genetic variation was detected among the 72 lines evaluated for quantitative and qualitative traits. Impressive transgressive segregation was measured for seed yield (22.42 g/plant), while plant height and maturity had higher heritabilities (73 and 89%, respectively). Other notable characters segregating in the population included panicle and stem color, panicle form, and resistance to downy mildew. In the Principal Component analysis, the first axis explained 74% of the total variation and was correlated to plant height, panicle size, stem diameter, biomass, mildew reaction, maturation, and seed yield; those traits are relevant discriminatory characters. Yield correlated positively with panicle length and biomass. Unweighted Pair Group Method with Arithmetic Mean-based cluster analysis identified three groups: one consisting of late, mildew-resistant, high-yielding lines; one having semi-late lines with intermediate yield and mildew susceptibility; and a third cluster consisting of early to semi-late accessions with low yield and mildew susceptibility. This study highlighted the extended diversity regenerated among the 72 accessions and helped to identify potentially adapted quinoa genotypes for production in the Moroccan coastal environment.

Keywords: quinoa, Chenopodium, downy mildew resistance, breeding, genetic diversity, Peronospora variabilis

## INTRODUCTION

fpls-07-01222 August 13, 2016 Time: 15:7 # 2

Quinoa (Chenopodium quinoa) is an ancient Andean seed crop of exceptional nutritional, and in particular protein, quality (Koziol, 1992). Its cultivation has expanded within the past decade beyond its traditional range to more than 70 countries (Food Agriculture Organization of United Nations [FAO], 2012), although the Andean nations of Peru and Bolivia are still primary quinoa producers. Due to quinoa's exceptional nutritional value, its capacity for adaptation to diverse agro-ecological conditions and its high commercial value, quinoa is expected to play an essential role in the UN-FAO strategy to sustainably feed the world's growing population (Jacobsen et al., 2013).

Biologically, C. quinoa belongs to an allotetraploid (2n = 4x = 36) complex found throughout the New World and whose root is the North American weed pitseed goosefoot (C. berlandieri). At least two other independent domestication events are presumed to have given rise to Mesoamerican vegetable and seed domesticates (C. berlandieri ssp. nuttaliae; Wilson and Heiser, 1979) as well as an extinct seed crop of the ancient cultures of eastern North America, C. berlandieri ssp. jonesianum (Smith and Yarnell, 2009; Kistler and Shapiro, 2011). After dispersing to the southeastern plains of South America, the weedy taxon of that region, avian goosefoot (C. hircinum), is presumed to have been brought into cultivation in the region of Lake Titicaca as C. quinoa and, at a later time, dispersed long-range to the Araucanian region on the Pacific slope in what is now Chile (Wilson, 1990; Bruno and Whitehead, 2003). Molecular genetic analyses have confirmed that the lowland Chilean coastal material is highly diverse (Fuentes et al., 2009), yet represents a branch that is separate from the even more variable quinoa germplasm of the High Andes (Christensen et al., 2007). The coastal Chilean germplasm includes genotypes that are highly resistant to heat and are day length insensitive – critical characters for quinoa breeders and agronomists seeking to expand quinoa's production into lowland subtropical and warm-season temperate environments around the world (Jacobsen et al., 2003).