Dissection of quantitative trait nucleotides and candidate genes associated with agronomic and yield-related traits under drought stress in rapeseed varieties: integration of genome-wide association study and transcriptomic analysis.

Introduction: An important strategy to combat yield loss challenge is the development of varieties with increased tolerance to drought to maintain production. Improvement of crop yield under drought stress is critical to global food security. Methods: In this study, we performed multiomics analysis in a collection of 119 diverse rapeseed (Brassica napus L.) varieties to dissect the genetic control of agronomic traits in two watering regimes [well-watered (WW) and drought stress (DS)] for 3 years. In the DS treatment, irrigation continued till the 50% pod development stage, whereas in the WW condition, it was performed throughout the whole growing season. Results: The results of the genome-wide association study (GWAS) using 52,157 single-nucleotide polymorphisms (SNPs) revealed 1,281 SNPs associated with traits. Six stable SNPs showed sequence variation for flowering time between the two irrigation conditions across years. Three novel SNPs on chromosome C04 for plant weight were located within drought tolerance-related gene ABCG16, and their pleiotropically effects on seed weight per plant and seed yield were characterized. We identified the C02 peak as a novel signal for flowering time, harboring 52.77% of the associated SNPs. The 288-kbps LD decay distance analysis revealed 2,232 candidate genes (CGs) associated with traits. The CGs BIG1-D, CAND1, DRG3, PUP10, and PUP21 were involved in phytohormone signaling and pollen development with significant effects on seed number, seed weight, and grain yield in drought conditions. By integrating GWAS and RNA-seq, 215 promising CGs were associated with developmental process, reproductive processes, cell wall organization, and response to stress. GWAS and differentially expressed genes (DEGs) of leaf and seed in the yield contrasting accessions identified BIG1-D, CAND1, and DRG3 genes for yield variation. Discussion: The results of our study provide insights into the genetic control of drought tolerance and the improvement of marker-assisted selection (MAS) for breeding high-yield and drought-tolerant varieties.

Transcriptome analysis in Aegilops tauschii unravels further insights into genetic control of stripe rust resistance.

MAIN CONCLUSION: The Aegilops tauschii resistant accession prevented the pathogen colonization by controlling the sugar flow and triggering the hypersensitive reaction. This study suggested that NBS-LRRs probably induce resistance through bHLH by controlling JA- and SA-dependent pathways. Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst) is one of wheat's most destructive fungal diseases that causes a severe yield reduction worldwide. The most effective and economically-friendly strategy to manage this disease is genetic resistance which can be achieved through deploying new and effective resistance genes. Aegilops tauschii, due to its small genome and co-evolution with Pst, can provide detailed information about underlying resistance mechanisms. Hence, we used RNA-sequencing approach to identify the transcriptome variations of two contrasting resistant and susceptible Ae. tauschii accessions in interaction with Pst and differentially expressed genes (DEGs) for resistance to stripe rust. Gene ontology, pathway analysis, and search for functional domains, transcription regulators, resistance genes, and protein-protein interactions were used to interpret the results. The genes encoding NBS-LRR, CC-NBS-kinase, and phenylalanine ammonia-lyase, basic helix-loop-helix (bHLH)-, basic-leucine zipper (bZIP)-, APETALA2 (AP2)-, auxin response factor (ARF)-, GATA-, and LSD-like transcription factors were up-regulated exclusively in the resistant accession. The key genes involved in response to salicylic acid, amino sugar and nucleotide sugar metabolism, and hypersensitive response contributed to plant defense against stripe rust. The activation of jasmonic acid biosynthesis and starch and sucrose metabolism pathways under Pst infection in the susceptible accession explained the colonization of the host. Overall, this study can fill the gaps in the literature on host-pathogen interaction and enrich the Ae. tauschii transcriptome sequence information. It also suggests candidate genes that could guide future breeding programs attempting to develop rust-resistant cultivars.

Genetic Variation of Puccinia triticina Populations in Iran from 2010 to 2017 as Revealed by SSR and ISSR Markers.

Puccinia triticina is a major wheat pathogen worldwide. Although Iran is within the Fertile Crescent, which is supposed to be the center of origin of both wheat and P. triticina, the knowledge of the genetic variability of local populations of this basidiomycete is limited. We analyzed 12 inter simple sequence repeats (ISSRs) and 18 simple sequence repeats (SSRs) of 175 P. triticina isolates sampled between 2010 and 2017 from wheat and other Poaceae in 14 provinces of Iran. SSRs revealed more polymorphisms than ISSRs, indicating they were more effective in differentiating P. triticina populations. Based on a dissimilarity matrix with a variable mutation rate for SSRs and a Dice coefficient for ISSRs, the isolates were separated into three large groups, each including isolates from diverse geographic origins and hosts. The grouping of SSR genotypes in UPGMA dendrograms was consistent with the grouping inferred from the Bayesian approach. However, isolates with a common origin clustered into separate subgroups within each group. The high proportion of heterozygous alleles suggests that in Iran clonal reproduction prevails over sexual reproduction of the pathogen. A significant correlation was found between SSR and ISSR genotypes and the virulence phenotypes of the isolates, as determined in a previous study.

Co-application of ACC deaminase-producing rhizobial bacteria and melatonin improves salt tolerance in common bean (Phaseolus vulgaris L.) through ion homeostasis.

A comprehensive body of scientific evidence indicates that rhizobial bacteria and melatonin enhance salt tolerance of crop plants. The overall goal of this research was to evaluate the ability of Rhizobium leguminoserum bv phaseoli to suppress salinity stress impacts in common bean treated with melatonin. Treatments included bacterial inoculations (inoculated (RI) and non-inoculated (NI)), different salinity levels (non-saline (NS), 4 (S1) and 8 (S2) dS m-1 of NaCl) and priming (dry (PD), melatonin (PM100) and hydro (PH) priming) with six replications in growing media containing sterile sand and perlite (1:1). The results showed that the bacterial strain had the ability to produce indole acetic acid (IAA), ACC deaminase and siderophore. Plants exposed to salinity stress indicated a significant decline in growth, yield, yield components, nitrogen fixation and selective transport (ST), while showed a significant increase in sodium uptake. However, the combination of PM100 and RI treatments by improving growth, photosynthesis rate and nitrogen fixation positively influenced plant performance in saline conditions. The combined treatment declined the negative impacts of salinity by improving the potassium translocation, potassium to sodium ratio in the shoot and root and ST. In conclusion, the combination of melatonin and ACC deaminase producing rhizobium mitigated the negative effects of salinity. This result is attributed to the increased ST and decreased sodium uptake, which significantly reduced the accumulation of sodium ions in shoot.

Foliar Application of Melatonin Improves the Salt Tolerance, Ion and Redox Homeostasis and Seed Oil Fatty Acid Profile in Camelina sativa.

Salinity affects the yield and quality of oilseed crops. The effects of a single foliar application of solutions with different concentrations (0, 30, 60 or 90 µM) of melatonin (MEL) to camelina (Camelina sativa) plants grown in soil in a greenhouse and irrigated at four salinity levels (0.5, 4, 8 and 16 dS m-1) were assessed. Increasing salinity decreased leaf chlorophyll and photosynthetic rates, decreased K concentrations and increased Na concentrations in roots and shoots, and increased oxidative marker levels and the activity of protective antioxidant enzymes in leaves. Under severe salinity stress, the MEL90 treatment resulted in increases in chlorophyll, gas exchange attributes, leaf antioxidant enzyme activities, and decreases in leaf oxidative markers and Na. Salinity decreased seed yield, with no seeds being produced at salinities above 8 dS m-1. The MEL90 treatment resulted in increases in seed yield and poly- and mono-unsaturated fatty acid contents and decreases in saturated fatty acid contents. The MEL90 treatment was more effective in alleviating salinity effects than those including lower MEL concentrations. The highest concentrations of K and K/Na ratios were observed with the MEL90 treatment under non-stressed conditions. Data suggest that MEL foliar applications could increase salinity stress tolerance in camelina.

Comparative transcriptome analysis of two contrasting resistant and susceptible Aegilops tauschii accessions to wheat leaf rust (Puccinia triticina) using RNA-sequencing.

Leaf rust, caused by Puccinia triticina Eriks., is the most common rust disease of wheat (Triticum aestivum L.) worldwide. Owing to the rapid evolution of virulent pathotypes, new and effective leaf rust resistance sources must be found. Aegilops tauschii, an excellent source of resistance genes to a wide range of diseases and pests, may provide novel routes for resistance to this disease. In this study, we aimed to elucidate the transcriptome of leaf rust resistance in two contrasting resistant and susceptible Ae. tauschii accessions using RNA-sequencing. Gene ontology, analysis of pathway enrichment and transcription factors provided an apprehensible review of differentially expressed genes and highlighted biological mechanisms behind the Aegilops-P. triticina interaction. The results showed the resistant accession could uniquely recognize pathogen invasion and respond precisely via reducing galactosyltransferase and overexpressing chromatin remodeling, signaling pathways, cellular homeostasis regulation, alkaloid biosynthesis pathway and alpha-linolenic acid metabolism. However, the suppression of photosynthetic pathway and external stimulus responses were observed upon rust infection in the susceptible genotype. In particular, this first report of comparative transcriptome analysis offers an insight into the strength and weakness of Aegilops against leaf rust and exhibits a pipeline for future wheat breeding programs.

Virulence of Leaf Rust Physiological Races in Iran From 2010 to 2017.

The wheat leaf rust fungus, Puccinia triticina, has widespread geographical distribution in Iran within the Fertile Crescent region of the Middle East where wheat was domesticated and P. triticina originated. Therefore, it is of great importance to identify the prevalence and distribution of P. triticina pathotypes in this area. From 2010 to 2017, 241 single-uredinium isolates of P. triticina were purified from 175 collections of P. triticina made from various hosts in 14 provinces of Iran, and they were tested on 20 Thatcher near-isogenic lines carrying single-leaf rust resistance genes. In total, 86 pathotypes were identified, of which the pathotypes FDTTQ, FDKPQ, FDKTQ, and FDTNQ were most prevalent. No virulence for Lr2a was detected, whereas virulence for Lr1 was found only on bread wheat in a few provinces in 2016. Only isolates from durum wheat and wild barley were virulent to Lr28. Although virulence for Lr9, Lr20, and Lr26 was observed in some years, the virulence frequency for these genes was lower than that of the other Lr genes. P. triticina collections from host plants with different ploidy levels or genetically dissimilar backgrounds were grouped individually according to genetic distance. Based on these results, collections from barley, durum wheat, oat, triticale, and wild barley were different from those of bread wheat.

Dissection of genotype × environment interactions for mucilage and seed yield in Plantago species: Application of AMMI and GGE biplot analyses.

Genotype × environment interaction (GEI) is an important aspect of both plant breeding and the successful introduction of new cultivars. In the present study, additive main effects and multiplicative interactions (AMMI) and genotype (G) main effects and genotype (G) × environment (E) interaction (GGE) biplot analyses were used to identify stable genotypes and to dissect GEI in Plantago. In total, 10 managed field trials were considered as environments to analyze GEI in thirty genotypes belonging to eight Plantago species. Genotypes were evaluated in a drought stress treatment and in normal irrigation conditions at two locations in Shiraz (Bajgah) for three years (2013-2014- 2015) and Kooshkak (Marvdasht, Fars, Iran) for two years (2014-2015). Three traits, seed yield and mucilage yield and content, were measured at each experimental site and in natural Plantago habitats. AMMI2 biplot analyses identified genotypes from several species with higher stability for seed yield and other genotypes with stable mucilage content and yield. P. lanceolata (G26), P. officinalis (G10), P. ovata (G14), P. ampleexcaulis (G11) and P. major (G4) had higher stability for seed yield. For mucilage yield, G21, G18 and G20 (P. psyllium), G1, G2 and G4 (P. major), G9 and G10 (P. officinalis) and P. lanceolata were identified as stable. G13 (P. ovata), G5 and G6 (P. major) and G30 (P. lagopus) had higher stability for mucilage content. No one genotype was found to have high levels of stability for more than one trait but some species had more than one genotype exhibiting stable trait performance. Based on trait variation, GGE biplot analysis identified two representative environments, one for seed yield and one for mucilage yield and content, with good discriminating ability. The identification of stable genotypes and representative environments should assist the breeding of new Plantago cultivars.

Microsatellite markers for the Triticum timopheevi-derived leaf rust resistance gene Lr18 on wheat 5BL chromosome.

Leaf rust, caused by Puccinia triticina, is a common wheat disease worldwide. Developing resistant cultivars through deploying new or pyramiding resistance genes in a suitable line, is the most effective approach to control this disease. However, to stack genes in a genotype, efficient and reliable markers are required. In the present study, F2 plants and their corresponding F3 families from a cross between the resistant line; Thatcher (Tc) Lr18, and the susceptible cultivar 'Boolani' were used to map rust resistance gene, Lr18 using SSR markers on chromosome 5BL of hexaploid wheat. The P. triticina pathotype no 15 was used to inoculate plants. Out of 20 primers tested, eight showed polymorphism between the two parents and were subsequently genotyped in the entire F2 population. The markers Xgpw7425 and Xwmc75 flanked the locus at a distance of 0.3 and 1.2 cM, respectively. Analysis of 81 genotypes from different backgrounds with these two markers confirmed their usefulness in screening absence or presence of Lr18. Therefore, these markers can be used for gene postulation and marker-assisted selection (MAS) of this gene in wheat breeding programs in future.