Quinoa Phenotyping Methodologies: An International Consensus

Quinoa is a crop originating in the Andes but grown more widely and with the genetic potential for significant further expansion. Due to the phenotypic plasticity of quinoa, varieties need to be assessed across years and multiple locations. To improve comparability among field trials across the globe and to facilitate collaborations, components of the trials need to be kept consistent, including the type and methods of data collected. Here, an internationally open-access framework for phenotyping a wide range of quinoa features is proposed to facilitate the systematic agronomic, physiological and genetic characterization of quinoa for crop adaptation and improvement. Mature plant phenotyping is a central aspect of this paper, including detailed descriptions and the provision of phenotyping cards to facilitate consistency in data collection. High-throughput methods for multi-temporal phenotyping based on remote sensing technologies are described. Tools for higher-throughput post-harvest phenotyping of seeds are presented. A guideline for approaching quinoa field trials including the collection of environmental data and designing layouts with statistical robustness is suggested. To move towards developing resources for quinoa in line with major cereal crops, a database was created. The Quinoa Germinate Platform will serve as a central repository of data for quinoa researchers globally.EEA FamailláFil: Stanschewski, Clara S. King Abdullah University of Science and Technology. Center for Desert Agriculture, Biological and Environmental Sciences and Engineering Division; Arabia SauditaFil: Rey, Elodie. King Abdullah University of Science and Technology. Center for Desert Agriculture, Biological and Environmental Sciences and Engineering Division; Arabia SauditaFil: Fiene, Gabriele. King Abdullah University of Science and Technology. Center for Desert Agriculture, Biological and Environmental Sciences and Engineering Division; Arabia SauditaFil: Craine, Evan B. Washington State University. Department of Crop and Soil Sciences; Estados UnidosFil: Wellman, Gordon. King Abdullah University of Science and Technology. Center for Desert Agriculture, Biological and Environmental Sciences and Engineering Division; Arabia SauditaFil: Melino, Vanessa J. King Abdullah University of Science and Technology. Center for Desert Agriculture, Biological and Environmental Sciences and Engineering Division; Arabia SauditaFil: Patiranage, Dilan S.R. King Abdullah University of Science and Technology. Center for Desert Agriculture, Biological and Environmental Sciences and Engineering Division; Arabia SauditaFil: Patiranage, Dilan S.R. Christian-Albrechts-University of Kiel. Plant Breeding Institute; AlemaniaFil: Johansen, Kasper. King Abdullah University of Science and Technology. Water Desalination and Reuse Center; Arabia SauditaFil: Schmöckel, Sandra M. University of Hohenheim. Institute of Crop Science. Department Physiology of Yield Stability; AlemaniaFil: Erazzu, Luis Ernesto. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Famaillá; Argentina.Fil: Tester, Mark. King Abdullah University of Science and Technology. Center for Desert Agriculture, Biological and Environmental Sciences and Engineering Division; Arabia Saudit

Quinoa phenotyping methodologies: An international consensus

Quinoa is a crop originating in the Andes but grown more widely and with the genetic potential for significant further expansion. Due to the phenotypic plasticity of quinoa, varieties need to be assessed across years and multiple locations. To improve comparability among field trials across the globe and to facilitate collaborations, components of the trials need to be kept consistent, including the type and methods of data collected. Here, an internationally open-access framework for phenotyping a wide range of quinoa features is proposed to facilitate the systematic agronomic, physiological and genetic characterization of quinoa for crop adaptation and improvement. Mature plant phenotyping is a central aspect of this paper, including detailed descriptions and the provision of phenotyping cards to facilitate consistency in data collection. High-throughput methods for multi-temporal phenotyping based on remote sensing technologies are described. Tools for higher-throughput post-harvest phenotyping of seeds are presented. A guideline for approaching quinoa field trials including the collection of environmental data and designing layouts with statistical robustness is suggested. To move towards developing resources for quinoa in line with major cereal crops, a database was created. The Quinoa Germinate Platform will serve as a central repository of data for quinoa researchers globally.Fil: Stanschewski, Clara S.. King Abdullah University of Science and Technology; Arabia SauditaFil: Rey, Elodie. King Abdullah University of Science and Technology; Arabia SauditaFil: Fiene, Gabriele. King Abdullah University of Science and Technology; Arabia SauditaFil: Craine, Evan B.. Washington State University; Estados UnidosFil: Wellman, Gordon. King Abdullah University of Science and Technology; Arabia SauditaFil: Melino, Vanessa J.. King Abdullah University of Science and Technology; Arabia SauditaFil: Patiranage, Dilan S. R.. King Abdullah University of Science and Technology; Arabia SauditaFil: Johansen, Kasper. King Abdullah University of Science and Technology; Arabia SauditaFil: Schmöckel, Sandra M.. King Abdullah University of Science and Technology; Arabia SauditaFil: Bertero, Hector Daniel. Universidad de Buenos Aires. Facultad de Agronomía. Departamento de Producción Vegetal. Cátedra de Producción Vegetal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; ArgentinaFil: Oakey, Helena. University of Adelaide; AustraliaFil: Colque Little, Carla. Universidad de Copenhagen; DinamarcaFil: Afzal, Irfan. University of Agriculture; PakistánFil: Raubach, Sebastian. The James Hutton Institute; Reino UnidoFil: Miller, Nathan. University of Wisconsin; Estados UnidosFil: Streich, Jared. Oak Ridge National Laboratory; Estados UnidosFil: Amby, Daniel Buchvaldt. Universidad de Copenhagen; DinamarcaFil: Emrani, Nazgol. Christian-albrechts-universität Zu Kiel; AlemaniaFil: Warmington, Mark. Agriculture And Food; AustraliaFil: Mousa, Magdi A. A.. Assiut University; Arabia Saudita. King Abdullah University of Science and Technology; Arabia SauditaFil: Wu, David. Shanxi Jiaqi Agri-Tech Co.; ChinaFil: Jacobson, Daniel. Oak Ridge National Laboratory; Estados UnidosFil: Andreasen, Christian. Universidad de Copenhagen; DinamarcaFil: Jung, Christian. Christian-albrechts-universität Zu Kiel; AlemaniaFil: Murphy, Kevin. Washington State University; Estados UnidosFil: Bazile, Didier. Savoirs, Environnement, Sociétés; Francia. Universite Paul-valery Montpellier Iii; FranciaFil: Tester, Mark. King Abdullah University of Science and Technology; Arabia Saudit

The Genome Sequence of the Wild Tomato Solanum pimpinellifolium Provides Insights Into Salinity Tolerance

Solanum pimpinellifolium, a wild relative of cultivated tomato, offers a wealth of breeding potential for desirable traits such as tolerance to abiotic and biotic stresses. Here, we report the genome assembly and annotation of S. pimpinellifolium ‘LA0480.’ Moreover, we present phenotypic data from one field experiment that demonstrate a greater salinity tolerance for fruit- and yield-related traits in S. pimpinellifolium compared with cultivated tomato. The ‘LA0480’ genome assembly size (811 Mb) and the number of annotated genes (25,970) are within the range observed for other sequenced tomato species. We developed and utilized the Dragon Eukaryotic Analyses Platform (DEAP) to functionally annotate the ‘LA0480’ protein-coding genes. Additionally, we used DEAP to compare protein function between S. pimpinellifolium and cultivated tomato. Our data suggest enrichment in genes involved in biotic and abiotic stress responses. To understand the genomic basis for these differences in S. pimpinellifolium and S. lycopersicum, we analyzed 15 genes that have previously been shown to mediate salinity tolerance in plants. We show that S. pimpinellifolium has a higher copy number of the inositol-3-phosphate synthase and phosphatase genes, which are both key enzymes in the production of inositol and its derivatives. Moreover, our analysis indicates that changes occurring in the inositol phosphate pathway may contribute to the observed higher salinity tolerance in ‘LA0480.’ Altogether, our work provides essential resources to understand and unlock the genetic and breeding potential of S. pimpinellifolium, and to discover the genomic basis underlying its environmental robustness

Validation of suitable genes for normalization of diurnal gene expression studies in Chenopodium quinoa.

Quinoa depicts high nutritional quality and abiotic stress resistance, attracting strong interest in the last years. To unravel the function of candidate genes for agronomically relevant traits, studying their transcriptional activities by RT-qPCR is an important experimental approach. The accuracy of such experiments strongly depends on precise data normalization. To date, validation of potential candidate genes for normalization of diurnal expression studies has not been performed in C. quinoa. We selected eight candidate genes based on transcriptome data and literature survey, including conventionally used reference genes. We used three statistical algorithms (BestKeeper, geNorm and NormFinder) to test their stability and added further validation by a simulation-based strategy. We demonstrated that using different reference genes, including those top ranked by stability, causes significant differences among the resulting diurnal expression patterns. Our results show that isocitrate dehydrogenase enzyme (IDH-A) and polypyrimidine tract-binding protein (PTB) are suitable genes to normalize diurnal expression data of two different quinoa accessions. Moreover, we validated our reference genes by normalizing two known diurnally regulated genes, BTC1 and BBX19. The validated reference genes obtained in this study will improve the accuracy of RT-qPCR data normalization and facilitate gene expression studies in quinoa

Identification of Putative Transmembrane Proteins Involved in Salinity Tolerance in Chenopodium quinoa by Integrating Physiological Data, RNAseq, and SNP Analyses

Chenopodium quinoa (quinoa) is an emerging crop that produces nutritious grains with the potential to contribute to global food security. Quinoa can also grow on marginal lands, such as soils affected by high salinity. To identify candidate salt tolerance genes in the recently sequenced quinoa genome, we used a multifaceted approach integrating RNAseq analyses with comparative genomics and topology prediction. We identified 219 candidate genes by selecting those that were differentially expressed in response to salinity, were specific to or overrepresented in quinoa relative to other Amaranthaceae species, and had more than one predicted transmembrane domain. To determine whether these genes might underlie variation in salinity tolerance in quinoa and its close relatives, we compared the response to salinity stress in a panel of 21 Chenopodium accessions (14 C. quinoa, 5 C. berlandieri, and 2 C. hircinum). We found large variation in salinity tolerance, with one C. hircinum displaying the highest salinity tolerance. Using genome re-sequencing data from these accessions, we investigated single nucleotide polymorphisms and copy number variation (CNV) in the 219 candidate genes in accessions of contrasting salinity tolerance, and identified 15 genes that could contribute to the differences in salinity tolerance of these Chenopodium accessions

Different NaCl-induced calcium signatures in the Arabidopsis thaliana ecotypes Col-0 and C24.

A common feature of stress signalling pathways are alterations in the concentration of cytosolic free calcium ([Ca2+]cyt), which allow the specific and rapid transmission of stress signals through a plant after exposure to a stress, such as salinity. Here, we used an aequorin based bioluminescence assay to compare the NaCl-induced changes in [Ca2+]cyt of the Arabidopsis ecotypes Col-0 and C24. We show that C24 lacks the NaCl specific component of the [Ca2+]cyt signature compared to Col-0. This phenotypic variation could be exploited as a screening methodology for the identification of yet unknown components in the early stages of the salt signalling pathway

Data from: Salinity tolerance loci revealed in rice using high-throughput non-invasive phenotyping

High-throughput phenotyping produces multiple measurements over time, which require new methods of analyses that are flexible in their quantification of plant growth and transpiration, yet are computationally economic. Here we develop such analyses and apply this to a rice population genotyped with a 700k SNP high-density array. Two rice diversity panels, indica and aus, containing a total of 553 genotypes, are phenotyped in waterlogged conditions. Using cubic smoothing splines to estimate plant growth and transpiration, we identify four time intervals that characterize the early responses of rice to salinity. Relative growth rate, transpiration rate and transpiration use efficiency (TUE) are analysed using a new association model that takes into account the interaction between treatment (control and salt) and genetic marker. This model allows the identification of previously undetected loci affecting TUE on chromosome 11, providing insights into the early responses of rice to salinity, in particular into the effects of salinity on plant growth and transpiration

Amplitude of the first [Ca²⁺]_cyt peak from luminometric measurements of Col-0 and C24 in response to [NaCl] and [sorbitol] treatment.

<p>The 13 d to 15 d old Col-0 (square symbol) and C24 (triangular symbol) seedlings constitutively expressing <i>aequorin</i> were treated with a range of NaCl and corresponding equal osmotic strength of sorbitol concentrations (which can be approximated to 2 times [NaCl]). Each point represents the average peak height of the first [Ca²⁺]_cyt peak of three replicates, error bars indicate the standard error of the mean (S.E.M.).</p

Jackknifed posterior probability that each individual stimulus belongs to an ecotype group defined in Fig. 2 B.

<p>Jackknifed posterior probability that each individual stimulus belongs to an ecotype group defined in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0117564#pone.0117564.g002" target="_blank">Fig. 2 B</a>.</p