Screening for drought tolerance and genetic diversity of wheat varieties using agronomic and molecular markers.

BACKGROUND: The future predictions for frequent and severe droughts will represent a significant threat to wheat yield and food security. In this context, breeding has proven to be the most efficient approach to enhance wheat productivity in dry environments. METHODS AND RESULTS: In this study, both agronomic and molecular-based approaches were used to evaluate the response of twenty-eight Tunisian wheat varieties to drought stress. The primary objective was to screen these varieties for drought tolerance using molecular and agro-morphological markers. All varieties were significantly affected by drought stress regarding various traits including total dry matter, straw length, flag leaf area, number of senescent leaves, SPAD value, grain yield and grain number. Furthermore, substantial variability in drought-stress tolerance was observed among wheat genotypes. The cluster analysis and principal component analyses confirmed the existence of genotypic variation in growth and yield impairments induced by drought. The stress susceptibility index (SSI) and tolerance index (TOL) proved to be the most effective indices and were strongly correlated with the varying levels of genotypic tolerance. The genotyping evaluation resulted in the amplification of 101 alleles using highly polymorphic 12 SSR markers, showed an average polymorphism of 74%. CONCLUSIONS: Taken together, the combination of agronomic and molecular approaches revealed that Karim, Td7, D117 and Utique are the most drought-tolerant wheat varieties. These varieties are particularly promising candidates for genetic improvements and can be utilized as potential genitors for future breeding programs in arid and semi-arid regions.

Drought responsiveness in six wheat genotypes: identification of stress resistance indicators.

Introduction: Wheat (Triticum aestivum L.) is among the world's most important staple food crops. In the current climate change scenario, a better understanding of wheat response mechanisms to water stress could help to enhance its productivity in arid ecosystems. Methods: In this study, water relations, gas exchange, membrane integrity, agronomic traits and molecular analysis were evaluated in six wheat genotypes (D117, Syndiouk, Tunisian durum7 (Td7), Utique, Mahmoudi AG3 and BT) subjected to drought-stress. Results and discussion: For all the studied genotypes, drought stress altered leaf area, chlorophyll content, stomatal density, photosynthetic rate and water-use efficiency, while the relative water content at turgor loss point (RWC0) remained stable. Changes in osmotic potential at turgor loss point (Ψπ0), bulk modulus of elasticity (Ɛmax) and stomatal regulation, differed greatly among the studied genotypes. For the drought-sensitive genotypes AG3 and BT, no significant changes were observed in Ψπ0, whereas the stomatal conductance (gs) and transpiration rate (E) decreased under stress conditions. These two varieties avoided turgor loss during drought treatment through an accurate stomatal control, resulting in a significant reduction in yield components. On the contrary, for Syndiouk, D117, Td7 and Utique genotypes, a solute accumulation and an increase in cell wall rigidity were the main mechanisms developed during drought stress. These mechanisms were efficient in enhancing soil water uptake, limiting leaf water loss and protecting cells membranes against leakage induced by oxidative damages. Furthermore, leaf soluble sugars accumulation was the major component of osmotic adjustment in drought-stressed wheat plants. The transcriptional analysis of genes involved in the final step of the ABA biosynthesis (AAO) and in the synthesis of an aquaporin (PIP2:1) revealed distinct responses to drought stress among the selected genotypes. In the resistant genotypes, PIP2:1 was significantly upregulated whereas in the sensitive ones, its expression showed only a slight induction. Conversely, the sensitive genotypes exhibited higher levels of AAO gene expression compared to the resistant genotypes. Our results suggest that drought tolerance in wheat is regulated by the interaction between the dynamics of leaf water status and stomatal behavior. Based on our findings, Syndiouk, D117, Utique and Td7, could be used in breeding programs for developing high-yielding and drought-tolerant wheat varieties.

Comparative Genetic Analysis of Durum Wheat Landraces and Cultivars Widespread in Tunisia.

The durum wheat (Triticum turgidum L. ssp. durum Desf.) landraces constitute a useful natural germplasm to increase the genetic diversity in the modern durum cultivars. The Tunisian durum germplasm constitutes 28 accessions conserved in Genebank of Tunisia, which are still unexplored. In this study, a comparative genetic analysis was performed to investigate the relationships between the Tunisian durum lines and the modern cultivars and detect divergent loci involved in breeding history. The genetic diversity analyses carried out using nine morphological descriptors and the 25K single-nucleotide polymorphism (SNP) array allowed us to distinguish two groups of Tunisian landraces and one of durum cultivars. The analysis of molecular variance and diversity indices confirmed the genetic variability among the groups. A total of 529 SNP loci were divergent between Tunisian durum landraces and modern cultivars. Candidate genes related to plant and spike architecture, including FLOWERING LOCUS T (FT-B1), zinc finger CONSTANS, and AP2/EREBPs transcription factors, were identified. In addition, divergent genes involved in grain composition and biotic stress nucleotide-binding site and leucine-reach repeats proteins and disease resistance proteins (NBS-LRR and RPM) were found, suggesting that the Tunisian durum germplasm may represent an important source of favorable alleles to be used in future durum breeding programs for developing well-adapted and resilient cultivars.

Serendipitous In Situ Conservation of Faba Bean Landraces in Tunisia: A Case Study.

Cultivation of faba bean (Vicia faba L.) in Tunisia is largely based on improved varieties of the crop. However, a few farmers continue to produce local cultivars or landraces. The National Gene Bank of Tunisia (NGBT) recently launched a collection project for faba bean landraces, with special focus on the regions of the North West, traditionally devoted to cultivating grain legumes, and where around 80% of the total national faba bean cultivation area is located. The seed phenotypic features of the collected samples were studied, and the genetic diversity and population structure analyzed using simple sequence repeat markers. The genetic constitution of the present samples was compared to that of faba bean samples collected by teams of the International Center for Agricultural Research in the Dry Areas (ICARDA) in the 1970s in the same region, and stored at the ICARDA gene bank. The results of the diversity analysis demonstrate that the recently collected samples and those stored at ICARDA largely overlap, thus demonstrating that over the past 50 years, little genetic change has occurred to the local faba bean populations examined. These findings suggest that farmers serendipitously applied international best practices for in situ conservation of agricultural crops.

Quality selection and genetic diversity of Tunisian durum wheat varieties using SSR markers / Seleção de qualidade e diversidade genética de vari edades de trigo duro da tunísia usando marcadores SSR

Our study focuses on the molecular analysis of the genetic diversity within 15 Tunisian durum wheat varieties and the assessment of the efficiency of some available markers to select valuable genotypes for technological proprieties of semolina (i.e. parameters related to SDS-sedimentation, mixing time and breakdown resistance of mixograph, grain protein content and yellow colour). While several markers were validated, others were not informative within the genotypes used. A high level of polymorphic information content (PIC) was detected, with an average of 5.2 polymorph alleles per locus and 0.6 average. Old varieties have high protein content however; modern varieties display strong gluten strength. Our results thus open the opportunity to choose valuable parents on the base of pedigrees, technological properties and genetic distances; and lead us to select efficient markers for the Regional Indigenous Land Strategy (Rils) selection strategy

Nosso estudo enfoca a análise molecular da diversidade genética em 15 variedades de trigo duro tunisiano e a avaliação da eficiência de alguns marcadores disponíveis para selecionar genótipos valiosos para propriedades tecnológicas de semolina (ou seja, parâmetros relacionados à sedimentação sds, tempo de mistura e resistência à degradação do mixógrafo, teor de proteína dos grãos e cor amarela). Enquanto vários marcadores foram validados, outros não foram informativos dentro dos genótipos utilizados. Foi detectado um alto nível de conteúdo de informação polimórfica (pic), com uma média de 5,2 alelos polimórficos por locus e 0,6 média. Variedades antigas têm alto teor de proteína no entanto; variedades modernas exibem forte força de glúten. Nossos resultados abrem, assim, a oportunidade de escolher pais valiosos com base em pedigrees, propriedades tecnológicas e distâncias genéticas; e nos levam a selecionar marcadores eficientes para a estratégia de seleção da estratégia regional de terras indígenas (rils).

Diversity of tunisian bread wheat genotypes revealed by morpho-agronomical and microsatellite markers / Diversidade de genótipos de trigo tunisiano para panificação obtida por marcadores morfo-agonômicos e microsatélites

The genetic diversity of a set of 21 hexaploid wheat germplasm from the National Agronomic Institute of Tunisia were investigated by applying 26 agro morphological traits and 10 wheat microsatellites molecular markers (Simple Sequence Repeat). The morphological variability was analyzed using the Principal Component Analysis (PCA) and the cluster analysis based on ward's method and square Euclidean distance. Eighteen microsatellites primer pairs were tested for all genotypes, among them 10 primers generated polymorphic and reproducible profiles. They revealed a total of 414 reducible bands among which 373 were polymorphic. The polymorphic information content (PIC) values per locus varied from 0,33 to 0,94 with an average of 0,72. Genetic similarity values between genotypes, calculated by the molecular derived data, were used to produce a dendrogram. The genotypes were clustered in four clear groups according to their origin, pedigree and in some cases to phenotypic characters similarities.

A diversidade genética de um conjunto de 21 hexaplóides em germoplasma de trigo oriundo do Instituto Nacional da Tunísia foi investigada pela aplicação de 26 caracteres morfológicos e 10 marcadores de microsatélites (Sequências simples repetidas). A variabilidade morfológica foi analisada pelo uso da análise de componentes principais (ACP) e análise de agrupamento baseada no método de Ward e o quadrado da distância Euclidiana. Dezoito primers de microsatélites foram testados para todos os genótipos, entre os quais 10 geraram polimorfismo e grupos distintos. Eles demonstraram um total de 414 bandas entre as quais 373 foram polimórficas. Os valores de polimorfismo encontrados por locus (VPL) variaram entre 0,33 a 0,94 com uma média de 0,72. Os valores da similaridade genética calculada para os dados moleculares originaram um dendograma. Os genótipos foram agrupados em quatro grupos de acordo com sua origem, pedigree (descendência) e em alguns casos pelos caracteres da similaridade fenotípica.