Heat Stress-Tolerant Quantitative Trait Loci Identified Using Backcrossed Recombinant Inbred Lines Derived from Intra-Specifically Diverse Aegilops tauschii Accessions.

In the face of climate change, bringing more useful alleles and genes from wild relatives of wheat is crucial to develop climate-resilient varieties. We used two populations of backcrossed recombinant inbred lines (BIL1 and BIL2), developed by crossing and backcrossing two intra-specifically diverse Aegilops tauschii accessions from lineage 1 and lineage 2, respectively, with the common wheat cultivar 'Norin 61'. This study aimed to identify quantitative trait loci (QTLs) associated with heat stress (HS) tolerance. The two BILs were evaluated under heat stress environments in Sudan for phenology, plant height (PH), grain yield (GY), biomass (BIO), harvest index (HI), and thousand-kernel weight (TKW). Grain yield was significantly correlated with BIO and TKW under HS; therefore, the stress tolerance index (STI) was calculated for these traits as well as for GY. A total of 16 heat-tolerant lines were identified based on GY and STI-GY. The QTL analysis performed using inclusive composite interval mapping identified a total of 40 QTLs in BIL1 and 153 QTLs in BIL2 across all environments. We detected 39 QTLs associated with GY-STI, BIO-STI, and TKW-STI in both populations (14 in BIL1 and 25 in BIL2). The QTLs associated with STI were detected on chromosomes 1A, 3A, 5A, 2B, 4B, and all the D-subgenomes. We found that QTLs were detected only under HS for GY on chromosome 5A, TKW on 3B and 5B, PH on 3B and 4B, and grain filling duration on 2B. The higher number of QTLs identified in BIL2 for heat stress tolerance suggests the importance of assessing the effects of intraspecific variation of Ae. tauschii in wheat breeding as it could modulate the heat stress responses/adaptation. Our study provides useful genetic resources for uncovering heat-tolerant QTLs for wheat improvement for heat stress environments.

Mining Aegilops tauschii genetic diversity in the background of bread wheat revealed a novel QTL for seed dormancy.

Due to the low genetic diversity in the current wheat germplasm, gene mining from wild relatives is essential to develop new wheat cultivars that are more resilient to the changing climate. Aegilops tauschii, the D-genome donor of bread wheat, is a great gene source for wheat breeding; however, identifying suitable genes from Ae. tauschii is challenging due to the different morphology and the wide intra-specific variation within the species. In this study, we developed a platform for the systematic evaluation of Ae. tauschii traits in the background of the hexaploid wheat cultivar 'Norin 61' and thus for the identification of QTLs and genes. To validate our platform, we analyzed the seed dormancy trait that confers resistance to preharvest sprouting. We used a multiple synthetic derivative (MSD) population containing a genetic diversity of 43 Ae. tauschii accessions representing the full range of the species. Our results showed that only nine accessions in the population provided seed dormancy, and KU-2039 from Afghanistan had the highest level of seed dormancy. Therefore, 166 backcross inbred lines (BILs) were developed by crossing the synthetic wheat derived from KU-2039 with 'Norin 61' as the recurrent parent. The QTL mapping revealed one novel QTL, Qsd.alrc.5D, associated with dormancy explaining 41.7% of the phenotypic variation and other five unstable QTLs, two of which have already been reported. The Qsd.alrc.5D, identified for the first time within the natural variation of wheat, would be a valuable contribution to breeding after appropriate validation. The proposed platform that used the MSD population derived from the diverse Ae. tauschii gene pool and recombinant inbred lines proved to be a valuable platform for mining new and important QTLs or alleles, such as the novel seed dormancy QTL identified here. Likewise, such a platform harboring genetic diversity from wheat wild relatives could be a useful source for mining agronomically important traits, especially in the era of climate change and the narrow genetic diversity within the current wheat germplasm.

Genetic variation for grain nutritional profile and yield potential in sorghum and the possibility of selection for drought tolerance under irrigated conditions.

BACKGROUND: Increasing grain nutritional value in sorghum (Sorghum bicolor) is a paramount breeding objective, as is increasing drought resistance (DR), because sorghum is grown mainly in drought-prone areas. The genetic basis of grain nutritional traits remains largely unknown. Marker-assisted selection using significant loci identified through genome-wide association study (GWAS) shows potential for selecting desirable traits in crops. This study assessed natural variation available in sorghum accessions from around the globe to identify novel genes or genomic regions with potential for improving grain nutritional value, and to study associations between DR traits and grain weight and nutritional composition. RESULTS: We dissected the genetic architecture of grain nutritional composition, protein content, thousand-kernel weight (TKW), and plant height (PH) in sorghum through GWAS of 163 unique African and Asian accessions under irrigated and post-flowering drought conditions. Several QTLs were detected. Some were significantly associated with DR, TKW, PH, protein, and Zn, Mn, and Ca contents. Genomic regions on chromosomes 1, 2, 4, 8, 9, and 10 were associated with TKW, nutritional, and DR traits; colocalization patterns of these markers indicate potential for simultaneous improvement of these traits. In African accessions, markers associated with TKW were mapped to six regions also associated with protein, Zn, Ca, Mn, Na, and DR, suggesting the potential for simultaneous selection for higher grain nutrition and TKW. Our results indicate that it may be possible to select for increased DR on the basis of grain nutrition and weight potential. CONCLUSIONS: This study provides a valuable resource for selecting landraces for use in plant breeding programs and for identifying loci that may contribute to grain nutrition and weight with the hope of producing cultivars that combine improved yield traits, nutrition, and DR.

Genome-wide association study reveals distinct genetic associations related to leaf hair density in two lineages of wheat-wild relative Aegilops tauschii.

Wild relatives of modern crops represent a promising source of genetic variation that can be mined for adaptations to climate change. Aegilops tauschii, the D-sub-genome progenitor of bread wheat (Triticum aestivum), constitutes a reservoir of genetic diversity for improving bread wheat performance and environmental resilience. Leaf hairiness plays an essential biological role in plant defense against biotic and abiotic stress. We investigated the natural variation in leaf hair density (LHD) among 293 Ae. tauschii accessions. Genome-wide association studies were performed for LHD with 2430 and 3880 DArTseq derived single nucleotide polymorphism (SNP) markers in two lineages of this species, TauL1 and TauL2, respectively. In TauL1, three marker-trait associations (MTAs) were located on chromosome 2D, whereas in TauL2, eight MTAs were identified, two associations were localized on each of the chromosomes 2D, 3D, 5D, and 7D. The markers explained phenotypic variation (R2) from 9 to 13% in TauL1 and 11 to 36% in TauL2. The QTLs identified in chromosomes 2D and 5D might be novel. Our results revealed more rapid and independent evolution of LHD in TauL2 compared to TauL1. The majority of LHD candidate genes identified are associated with biotic and abiotic stress responses. This study highlights the significance of intraspecific diversity of Ae. tauschii to enhance cultivated wheat germplasm.

Identification of Glu-D1 Alleles and Novel Marker-Trait Associations for Flour Quality and Grain Yield Traits under Heat-Stress Environments in Wheat Lines Derived from Diverse Accessions of Aegilops tauschii.

Heat stress during grain filling is considered one of the major abiotic factors influencing wheat grain yield and quality in arid and semi-arid regions. We studied the effect of heat stress on flour quality and grain yield at moderate and continuous heat stress under natural field conditions using 147 lines of wheat multiple synthetic derivatives (MSD) containing Aegilops tauschii introgressions. The study aimed to identify the marker-trait associations (MTAs) for the quality traits and grain yield under heat-stress conditions and identify stress-resilient germplasm-combining traits for good flour quality and grain yield. The MSD lines showed considerable genetic variation for quality traits and grain yield under heat-stress conditions; some lines performed better than the recurrent parent, Norin 61. We identified two MSD lines that consistently maintained relative performance (RP) values above 100% for grain yield and dough strength. We found the presence of three high-molecular-weight glutenin subunits (HMW-GSs) at the Glu-D1 locus derived from Ae. tauschii, which were associated with stable dough strength across the four environments used in this study. These HMW-GSs could be potentially useful in applications for future improvements of end-use quality traits targeting wheat under severe heat stress. A total of 19,155 high-quality SNP markers were used for the genome-wide association analysis and 251 MTAs were identified, most of them on the D genome, confirming the power of the MSD panel as a platform for mining and exploring the genes of Ae. tauschii. We identified the MTAs for dough strength under heat stress, which simultaneously control grain yield and relative performance for dough strength under heat-stress/optimum conditions. This study proved that Ae. tauschii is an inexhaustible resource for genetic mining, and the identified lines and pleiotropic MTAs reported in this study are considered a good resource for the development of resilient wheat cultivars that combine both good flour quality and grain yield under stress conditions using marker-assisted selection.

Exploiting Wild Emmer Wheat Diversity to Improve Wheat A and B Genomes in Breeding for Heat Stress Adaptation.

Wheat is highly sensitive to temperature beyond the optimum. To improve wheat adaptation to heat stress, the best option is to exploit the diversity of wild wheat progenitors. This study aimed to identify germplasm and quantitative trait loci associated with heat stress tolerance from wild emmer wheat diversity. We evaluated a diverse set of multiple derivative lines harboring chromosome segments from nine wild emmer wheat parents under four environments: two optimum environments at Tottori, Japan and Dongola, Sudan, one moderate heat stress environment, and one severe heat stress environment at Wad Medani, Sudan. Genome-wide association analysis was conducted with 13,312 SNP markers. Strong marker-trait associations (MTAs) were identified for chlorophyll content at maturity on chromosomes 1A and 5B: these MTAs explained 28.8 and 26.8% of the variation, respectively. A region on chromosome 3A (473.7-638.4 Mbp) contained MTAs controlling grain yield, under optimum and severe heat stress. Under severe heat stress, regions on chromosomes 3A (590.4-713.3 Mbp) controlled grain yield, biomass, days to maturity and thousand kernel weight, and on 3B (744.0-795.2 Mbp) grain yield and biomass. Heat tolerance efficiency (HTE) was controlled by three MTAs, one each on chromosomes 2A, 2B, and 5A under moderate heat stress and one MTA on chromosome 3A under severe heat stress. Some of the MTAs found here were previously reported, but the new ones originated from the wild emmer wheat genomes. The favorable alleles identified from wild emmer wheat were absent or rare in the elite durum wheat germplasm being bred for heat stress tolerance. This study provides potential genetic materials, alleles, MTAs, and quantitative trait loci for enhancing wheat adaptation to heat stress. The derivative lines studied here could be investigated to enhance other stress tolerance such as drought and salinity.

Harnessing the diversity of wild emmer wheat for genetic improvement of durum wheat.

KEY MESSAGE: The multiple derivative lines (MDLs) characterized in this study offer a promising strategy for harnessing the diversity of wild emmer wheat for durum and bread wheat improvement. Crop domestication has diminished genetic diversity and reduced phenotypic plasticity and adaptation. Exploring the adaptive capacity of wild progenitors offer promising opportunities to improve crops. We developed a population of 178 BC1F6 durum wheat (Triticum turgidum ssp. durum) lines by crossing and backcrossing nine wild emmer wheat (T. turgidum ssp. dicoccoides) accessions with the common durum wheat cultivar 'Miki 3'. Here, we describe the development of this population, which we named as multiple derivative lines (MDLs), and demonstrated its suitability for durum wheat breeding. We genotyped the MDL population, the parents, and 43 Sudanese durum wheat cultivars on a Diversity Array Technology sequencing platform. We evaluated days to heading and plant height in Dongola (Sudan) and in Tottori (Japan). The physical map length of the MDL population was 9 939 Mb with an average of 1.4 SNP/Mb. The MDL population had greater diversity than the Sudanese cultivars. We found high gene exchange between the nine wild emmer accessions and the MDL population, indicating that the MDL captured most of the diversity in the wild emmer accessions. Genome-wide association analysis identified three loci for days to heading on chromosomes 1A and 5A in Dongola and one on chromosome 3B in Tottori. For plant height, common genomic loci were found on chromosomes 4A and 4B in both locations, and one genomic locus on chromosome 7B was found only in Dongola. The results revealed that the MDLs are an effective strategy towards harnessing wild emmer wheat diversity for wheat genetic improvement.

Tuning Beforehand: A Foresight on RNA Interference (RNAi) and In Vitro-Derived dsRNAs to Enhance Crop Resilience to Biotic and Abiotic Stresses.

Crop yield is severely affected by biotic and abiotic stresses. Plants adapt to these stresses mainly through gene expression reprogramming at the transcriptional and post-transcriptional levels. Recently, the exogenous application of double-stranded RNAs (dsRNAs) and RNA interference (RNAi) technology has emerged as a sustainable and publicly acceptable alternative to genetic transformation, hence, small RNAs (micro-RNAs and small interfering RNAs) have an important role in combating biotic and abiotic stresses in plants. RNAi limits the transcript level by either suppressing transcription (transcriptional gene silencing) or activating sequence-specific RNA degradation (post-transcriptional gene silencing). Using RNAi tools and their respective targets in abiotic stress responses in many crops is well documented. Many miRNAs families are reported in plant tolerance response or adaptation to drought, salinity, and temperature stresses. In biotic stress, the spray-induced gene silencing (SIGS) provides an intelligent method of using dsRNA as a trigger to silence target genes in pests and pathogens without producing side effects such as those caused by chemical pesticides. In this review, we focus on the potential of SIGS as the most recent application of RNAi in agriculture and point out the trends, challenges, and risks of production technologies. Additionally, we provide insights into the potential applications of exogenous RNAi against biotic stresses. We also review the current status of RNAi/miRNA tools and their respective targets on abiotic stress and the most common responsive miRNA families triggered by stress conditions in different crop species.

Exploitation of Tolerance of Wheat Kernel Weight and Shape-Related Traits from Aegilops tauschii under Heat and Combined Heat-Drought Stresses.

Kernel weight and shape-related traits are inherited stably and increase wheat yield. Narrow genetic diversity limits the progress of wheat breeding. Here, we evaluated kernel weight and shape-related traits and applied genome-wide association analysis to a panel of wheat multiple synthetic derivative (MSD) lines. The MSD lines harbored genomic fragments from Aegilops tauschii. These materials were grown under optimum conditions in Japan, as well as under heat and combined heat-drought conditions in Sudan. We aimed to explore useful QTLs for kernel weight and shape-related traits under stress conditions. These can be useful for enhancing yield under stress conditions. MSD lines possessed remarkable genetic variation for all traits under all conditions, and some lines showed better performance than the background parent Norin 61. We identified 82 marker trait associations (MTAs) under the three conditions; most of them originated from the D genome. All of the favorable alleles originated from Ae. tauschii. For the first time, we identified markers on chromosome 5D associated with a candidate gene encoding a RING-type E3 ubiquitin-protein ligase and expected to have a role in regulating wheat seed size. Our study provides important knowledge for the improvement of wheat yield under optimum and stress conditions. The results emphasize the importance of Ae. tauschii as a gene reservoir for wheat breeding.

Genome-Wide Association Study of Morpho-Physiological Traits in Aegilops tauschii to Broaden Wheat Genetic Diversity.

Aegilops tauschii, the D-genome donor of bread wheat, is a storehouse of genetic diversity that can be used for wheat improvement. This species consists of two main lineages (TauL1 and TauL2) and one minor lineage (TauL3). Its morpho-physiological diversity is large, with adaptations to a wide ecological range. Identification of allelic diversity in Ae. tauschii is of utmost importance for efficient breeding and widening of the genetic base of wheat. This study aimed at identifying markers or genes associated with morpho-physiological traits in Ae. tauschii, and at understanding the difference in genetic diversity between the two main lineages. We performed genome-wide association studies of 11 morpho-physiological traits of 343 Ae. tauschii accessions representing the entire range of habitats using 34,829 DArTseq markers. We observed a wide range of morpho-physiological variation among all accessions. We identified 23 marker-trait associations (MTAs) in all accessions, 15 specific to TauL1 and eight specific to TauL2, suggesting independent evolution in each lineage. Some of the MTAs could be novel and have not been reported in bread wheat. The markers or genes identified in this study will help reveal the genes controlling the morpho-physiological traits in Ae. tauschii, and thus in bread wheat even if the plant morphology is different.

Exploiting the potential of Sudanese sorghum landraces in biofortification: Physicochemical quality of the grain of sorghum (Sorghum bicolor L. Moench) landraces.

This study aimed to describe the phytonutrients and antioxidant activity, protein content, in vitro protein digestibility (IVPD), protein fraction, and bioavailability of Fe and Zn in the grains of five sorghum landraces grown in Sudan. The results showed significant differences in all quality tests among the landraces. The Tetron landrace showed the highest percentage of crude protein and IVPD among the landraces. Additionally, most of the landrace grains had high contents of Fe and Zn with a high rate of bioavailability. The Kolom 4055 and Wad akar exhibited significantly higher total phenolic contents, with antioxidant activity of 79.3% and 83.4%, respectively. The glutelin content was relatively higher compared to the other fractions, irrespective of sorghum landraces. The principal components cumulatively accounted for 89.3% of the total variation among the five sorghum landraces. It can be concluded that these landraces could be used in the improvement of new value-added crops using the by-products of sorghum grains.

Stay-Green Trait: A Prospective Approach for Yield Potential, and Drought and Heat Stress Adaptation in Globally Important Cereals.

The yield losses in cereal crops because of abiotic stress and the expected huge losses from climate change indicate our urgent need for useful traits to achieve food security. The stay-green (SG) is a secondary trait that enables crop plants to maintain their green leaves and photosynthesis capacity for a longer time after anthesis, especially under drought and heat stress conditions. Thus, SG plants have longer grain-filling period and subsequently higher yield than non-SG. SG trait was recognized as a superior characteristic for commercially bred cereal selection to overcome the current yield stagnation in alliance with yield adaptability and stability. Breeding for functional SG has contributed in improving crop yields, particularly when it is combined with other useful traits. Thus, elucidating the molecular and physiological mechanisms associated with SG trait is maybe the key to defeating the stagnation in productivity associated with adaptation to environmental stress. This review discusses the recent advances in SG as a crucial trait for genetic improvement of the five major cereal crops, sorghum, wheat, rice, maize, and barley with particular emphasis on the physiological consequences of SG trait. Finally, we provided perspectives on future directions for SG research that addresses present and future global challenges.

Nutritional potential of wild sorghum: Grain quality of Sudanese wild sorghum genotypes (Sorghum bicolor L. Moench).

In the last decades, deficiency of macro- and micronutrients was considered as a serious problem associated with the increase in the human population. To meet the increased demand for food consumption, the wild relative plant might serve as an important source of new genetic material for increasing macro- and micronutrients. To investigate this, the variations in protein content, in vitro protein digestibility, tannin content, phytic acid content, total polyphenol content, and total and bioavailability of minerals were studied in grains of ten wild sorghums and two released sorghum cultivars. The results showed significant differences (p ≤ 0.05) in all quality tests among the genotypes. The highest percentage of total protein contents and in vitro protein digestibility were encountered in the grains of PQ-434 (14.6%) and the released cultivar AG8 (49.8%), respectively, while the highest concentrations of total and bioavailable iron were found in the grains of Almahkara (3.17 mg/100 g) and Abusabiba (92.8 mg/100 g), respectively. The grains of wild sorghum genotype Adar Umbatikh grains were found to possess higher total zinc contents. The PCA identified only five components of eigenvalues greater than one and cumulatively accounted for 88% of the total variation. It could be concluded that Almahkara and PQ-434 could be used as potential sources for iron and protein sorghum biofortification, respectively. Results from this study might be used in the development of new value-added products from wild sorghum grains by-products.

Stay-Green QTLs Response in Adaptation to Post-Flowering Drought Depends on the Drought Severity.

Stay-green trait enhances sorghum adaptation to post-flowering drought. Six stay-green backcross introgression lines (BILs) carrying one or more stay-green QTLs (Stg1-4) and their parents were characterized under non-stress (W100: 100% of soil field capacity (FC)) and two levels of post-flowering drought (W75: 75% FC; W50: 50% FC) in a controlled condition. We aimed to study the response and identify the drought threshold of these QTLs under different levels of post-flowering drought and find traits closely contributing to grain yield (GY) under different drought severity. W50 caused the highest reduction in BILs performance. From W100 to W50, the GY of the recurrent parent reduced by 70%, whereas that of the BILs reduced by only 36%. W75 and W50 induce different behavior/response compared to W100. Harvest index contributed to the GY under the three water regimes. For high GY under drought transpiration rate at the beginning of drought and mid-grain filling was important at W75, whereas it was important at mid-grain filling and late-grain filling at W50. Stay-green trait can be scored simply with the relative number of green leaves/plants under both irrigated and stress environments. QTL pyramiding might not always be necessary to stabilize or increase the GY under post-flowering drought. The stay-green QTLs increase GY under drought by manipulating water utilization depending on drought severity.

Impact of wheat-Leymus racemosus added chromosomes on wheat adaptation and tolerance to heat stress.

Adaptation of wheat (Triticum aestivum L.) to high temperatures could be improved by introducing alien genes from wild relatives. We evaluated the responses of wheat-Leymus racemosus chromosome introgression lines to high temperature to determine their potentiality for developing improved wheat cultivars. Introgression lines and their parent Chinese Spring were evaluated in a growth chamber at the seedling stage and in the field at the reproductive stage in two heat-stressed environments in Sudan. Optimum and late planting were used to ensure exposure of the plants to heat stress at the reproductive stage. The results revealed the impact of several Leymus chromosomes in improving wheat adaptation and tolerance to heat. Three lines possessed enhanced adaptation, whereas two showed high heat tolerance. Two addition lines showed a large number of kernels per spike, while one possessed high yield potential. Grain yield was correlated negatively with the heat susceptibility index, days to heading and maturity and positively with kernel number per spike and triphenyl tetrazolium chloride assay under late planting. The findings suggest that these genetic stocks could be used as a bridge to introduce the valuable Leymus traits into a superior wheat genetic background, thus helping maximize wheat yield in heat-stressed environments.