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Background Leaf rust, caused by Puccinia triticina Eriks ( Pt ) is a major threat to wheat cultivation worldwide. The rapid evolution of this pathogen has led to the emergence of new virulent strains that can overcome the resistance of commonly cultivated wheat varieties. To address this threat, continuous monitoring of leaf rust pathotypes is conducted in wheat-growing regions across the world. This approach helps prioritize the development and deployment of resistant cultivars, as well as the implementation of other effective control measures against the prevailing races. The key wheat leaf rust pathotypes in India include 77–5 (121R63-1), 77–6 (121R55-1), 77–9 (121R60-1), 12–5 (29R45), and 104 (17R23). Among these pathotypes, 77–5 (121R63-1) and 77–9 (121R60-1) are the most prevalent since 2016. As virulent pathotypes continue to evolve and adapt, there is an urgent need to continually explore the vast germplasm repositories of wheat and its related species to identify novel genetic resources and genes that confer resistance to these evolving leaf rust pathotypes. Therefore, the present study aims to identify genes and genomic regions responsible for leaf rust resistance against prevalent pathotypes in India, focusing on a subset of the Global Durum Wheat Panel, which includes genotypes from various tetraploid wheat species. Results This study revealed wide variation in seedling-stage resistance among 189 tetraploid wheat accessions against five prevalent leaf rust pathotypes in India namely, 77–5 (121R63-1), 77–6 (121R55-1), 77–9 (121R60-1), 12–5 (29R45) and 104 (17R23). Approximately 45% of the population exhibited immune/highly resistant to moderately resistant responses to pathotypes 77–5, 77–6 and 104, while around 23–27% showed similar levels of resistance to pathotypes 77–9 and 12–5. A genome-wide association study using six multi-locus models identified 88 significantly associated quantitative trait nucleotides (QTNs) across the five leaf rust pathotypes. Among these, 22 QTNs were considered reliable, including four for pathotype 77–5, six for 12–5, three for 77–9, seven for 104, and two for 77–6. Among the 22 reliable QTNs, 10 coincided with the rust resistance regions reported in previous studies, whereas 12 appeared to be novel. Further investigations of the regions flanking all 88 QTNs revealed 300 genes, including 62 associated with disease resistance or defense responses. In silico expression analysis of these defense-related genes revealed two nucleotide-binding site–leucine-rich repeat genes: one on chromosome 6B ( TRITD6Bv1G224600 ) near QTN RAC875_c35430_373 , and another on chromosome 6A ( TRITD6Av1G225060 ) in the vicinity of QTN Excalibur_c77841_224 with significantly higher levels of expression in the leaf-resistant genotype during the early hours of Pt infection. Therefore, these two genes could be potential candidates for resistance to leaf rust in tetraploid wheat germplasm. Conclusions Our study provides a comprehensive understanding of the genetic basis underlying leaf rust resistance in a diverse tetraploid wheat germplasm panel. It has also revealed novel candidate genomic regions for leaf rust resistance. These genomic regions represent important targets for inclusion in marker-assisted breeding initiatives, aimed at fostering durable resistance against leaf rust disease.

Zinc finger-homeodomain (ZF-HDs) class IV transcriptional factors (TFs) is a plant-specific transcription factor and play a key role in stress responses, plant growth, development, and hormonal signaling. In this study, two new leaf rolling TFs genes, namely TaZHD1 and TaZHD10 , were identified in wheat using comparative genomic analysis of the target region that carried a major QTL for leaf rolling identified through multi-environment phenotyping and high throughput genotyping of a RIL population. Structural and functional annotation of the candidate ZHD genes with its closest rice orthologs reflects the species-specific evolution and, undoubtedly, validates the notions of remote-distance homology concept. Meanwhile, the morphological analysis resulted in contrasting difference for leaf rolling in extreme RILs between parental lines HD2012 and NI5439 at booting and heading stages. Transcriptome-wide expression profiling revealed that TaZHD10 transcripts showed significantly higher expression levels than TaZHD1 in all leaf tissues upon drought stress. The relative expression of these genes was further validated by qRT-PCR analysis, which also showed consistent results across the studied genotypes at the booting and anthesis stage. The contrasting modulation of these genes under drought conditions and the available evidenced for its epigenetic behavior that might involve the regulation of metabolic and gene regulatory networks. Prediction of miRNAs resulted in five Tae-miRs that could be associated with RNAi mediated control of TaZHD1 and TaZHD10 putatively involved in the metabolic pathway controlling rolled leaf phenotype. Gene interaction network analysis indicated that TaZHD1 and TaZHD10 showed pleiotropic effects and might also involve other functions in wheat in addition to leaf rolling. Overall, the results increase our understanding of TaZHD genes and provide valuable information as robust candidate genes for future functional genomics research aiming for the breeding of wheat varieties tolerant to leaf rolling.

Hybrid development is one of the most promising strategies for boosting crop yields. Parental lines used to create hybrids must have good per se performance and disease resistance for developing superior hybrids. Indian wheat line HD3209 was developed by introducing the rust resistance genes Lr19/Sr25 into the background of popular wheat variety HD2932. The wheat line HD3209 carrying Lr19/Sr25 has been successfully and rapidly converted to the CMS line A-HD3209, with 96.01% background genome recovery, based on selection for agro-morphological traits, rust resistance, pollen sterility, and foreground and background analyses utilizing SSR markers. The converted CMS line A-HD3209 was completely sterile and nearly identical to the recurrent parent HD3209. Based on high per se performance and rust resistance, the study concludes that the derived CMS line A-HD3209 is promising and can be employed successfully in hybrid development.

Salinity is a major abiotic stress in wheat production across the globe, especially in arid and semi-arid areas. In this study, 313 genetically diverse wheat genotypes were assessed for vegetative-stage salinity tolerance in a hydroponic condition and genotyped using 35K Axiom SNP array. Genotyping of the 313 wheat genotypes produced 24,968 polymorphic SNPs. To dissect salt tolerance, marker-trait association analysis was carried out using the salt tolerance indices of three traits, such as leaf chlorophyll content, green leaf area, and dry biomass. In total, 24 quantitative trait nucleotides (QTNs) showing significant associations with these three traits were identified, including seven linked to leaf chlorophyll content, twelve with green leaf area, and five to dry biomass traits. Four QTNs (Q.CCI-E3-1A, Q.CCI-E3-5D, Q.DB-E3-1B, and Q.GLA-E3-5A.2) showed significant phenotypic effects and represented GWAS-significant loci under both control and salt-stress conditions. Gene ontology analysis of the genomic regions linked to these QTNs revealed 67 putative candidate genes associated with ion transport, stress signaling, and photosynthetic processes. The identified SNPs, QTNs, and candidate genes provide valuable genomic resources for marker-assisted breeding of salt-tolerant wheat cultivars, contributing to sustainable wheat production under saline environments.

Background Wheat plays a pivotal role in global food and nutritional security. To meet the growing demand for food, increasing wheat production through hybrid development remains an untapped avenue. However, the autogamy of wheat causes a significant challenge for hybrid development. Results The present study aimed to convert the elite bread wheat cultivars HD3086 and HD2932 into a cytoplasmic male-sterile (CMS) lines using the CMS donor parent (A-GW365) through a backcross breeding approach. Background analysis using 152 and 145 SSR markers confirmed ˃95% recovery of recurrent parent genomes (RPG) of the HD3086 and HD2932, respectively. The newly developed CMS lines were evaluated for pollen sterility and phenotypic similarity in comparison to recurrent parents. The cytological study and DUS characterisation of the converted A lines revealed complete sterility and similarity with the recurrent parent for morphological and agronomic traits. Further, two converted A lines, A-HD3086 and A-HD2932 and donor A line A-GW365 were crossed with six newly developed fertility restorer lines (R lines) in a line × tester breeding design. Combining ability analysis revealed positive general combining ability (GCA) for A-HD3086 and 955R across the three trials, and they were identified as the best tester and line, respectively, for grain yield. Furthermore, the genotype × environment interaction analysed through GGE biplot revealed that hybrids G1 (A-HD3086 × 908-3R), G2 (A-HD3086 × 917R), G4 (A-HD3086 × 955R), and G12 (A-GW365 × 1752R) were high-yielding and stable performers. Based on combining ability estimates, grain yield performance, and stability analysis, hybrids G4 (A-HD3086 × 955R) and G12 (A-GW365 × 1752R) were identified as the best-performing hybrids across the environmental trials. Conclusions The present study reported the conversion of agronomically superior cultivars to CMS lines and their practical utilization for the development of CMS-based hybrids.

Guava ( Psidium guajava L.) is one of the economically major fruit crops, abundant in nutrients and found growing in tropical-subtropical regions around the world. Ensuring sufficient genomic resources is crucial for crop species to enhance breeding efficiency and facilitate molecular breeding. However, genomic resources, especially microsatellite or simple sequence repeat (SSR) markers, are limited in guava. Therefore, novel genome-wide SSR markers were developed by utilizing chromosome assembly (GCA_016432845.1) of the “New Age” cultivar through GMATA, a comprehensive software. The software evaluated about 397.8 million base pairs (Mbp) of the guava genome sequence, where 87,372 SSR loci were utilized to design primers, ultimately creating 75,084 new SSR markers. After in silico analysis, a total of 75 g-SSR markers were chosen to screen 35 guava genotypes, encompassing wild Psidium species and five jamun genotypes. Of the 72 amplified novel g-SSR markers (FHTGSSRs), 53 showed polymorphism, suggesting significant genetic variation among the guava genotypes, including wild species. The 53 polymorphic g-SSR markers had an average of 3.04 alleles per locus for 35 selected guava genotypes. Besides, in this study, the mean values recorded for major allele frequency, gene diversity, observed heterozygosity, and polymorphism information content were 0.73, 0.38, 0.13, and 0.33, respectively. Among the wild Psidium species studied, the transferability of these novel g-SSR loci across different species was found to be 45.83% to 90.28%. Furthermore, 17 novel g-SSR markers were successfully amplified in all the selected Syzygium genotypes, of which only four markers could differentiate between two Syzygium species. A neighbour-joining (N-J) tree was constructed using 53 polymorphic g-SSR markers and classified 35 guava genotypes into four clades and one outlier, emphasizing the genetic uniqueness of wild Psidium species compared to cultivated genotypes. Model-based structure analysis divided the guava genotypes into two distinct genetic groups, a classification that was strongly supported by Principal Coordinate Analysis (PCoA). In addition, the AMOVA and PCoA analyses also indicated substantial genetic diversity among the selected guava genotypes, including wild Psidium species. Hence, the developed novel genome-wide genomic SSRs could enhance the availability of genomic resources and assist in the molecular breeding of guava.

Wheat leaf rust caused by Puccinia triticina Eriks is an important disease that causes yield losses of up to 40% in susceptible varieties. Tetraploid emmer wheat (T. turgidum ssp. Dicoccum), commonly called Khapli wheat in India, is known to have evolved from wild emmer (Triticum turgidum var. dicoccoides), and harbors a good number of leaf rust resistance genes. In the present study, we are reporting on the screening of one hundred and twenty-three dicoccum wheat germplasm accessions against the leaf rust pathotype 77-5. Among these, an average of 45.50% of the germplasms were resistant, 46.74% were susceptible, and 8.53% had mesothetic reactions. Further, selected germplasm lines with accession numbers IC138898, IC47022, IC535116, IC535133, IC535139, IC551396, and IC534144 showed high level of resistance against the eighteen prevalent pathotypes. The infection type varied from “;”, “;N”, “;N1” to “;NC”. PCR-based analysis of the resistant dicoccum lines with SSR marker gwm508 linked to the Lr53 gene, a leaf rust resistance gene effective against all the prevalent pathotypes of leaf rust in India and identified from a T. turgidum var. dicoccoides germplasm, indicated that Lr53 is not present in the selected accessions. Moreover, we have also generated 35K SNP genotyping data of seven lines and the susceptible control, Mandsaur Local, to study their relationships. The GDIRT tool based on homozygous genotypic differences revealed that the seven genotypes are unique to each other and may carry different resistance genes for leaf rust.

The F-box proteins in fungi perform diverse functions including regulation of cell cycle, circadian clock, development, signal transduction and nutrient sensing. Genome-wide analysis revealed 10 F-box genes in Puccinia triticina, the causal organism for the leaf rust disease in wheat and were characterized using in silico approaches for revealing phylogenetic relationships, gene structures, gene ontology, protein properties, sequence analysis and gene expression studies. Domain analysis predicted functional domains like WD40 and LRR at C-terminus along with the obvious presence of F-box motif in N-terminus. MSA showed amino acid replacements, which might be due to nucleotide substitution during replication. Phylogenetic analysis revealed the F-box proteins with similar domains to be clustered together while some sequences were spread out in different clades, which might be due to functional diversity. The clustering of Puccinia triticina GG705409 with Triticum aestivum TaAFB4/TaAFB5 in a single clade suggested the possibilities of horizontal gene transfer during the coevolution of P. triticina and wheat. Gene ontological annotation categorized them into three classes and were functionally involved in protein degradation through the protein ubiquitination pathway. Protein–protein interaction network revealed F-box proteins to interact with other components of the SCF complex involved in protein ubiquitination. Relative expression analysis of five F-box genes in a time course experiment denoted their involvement in leaf rust susceptible wheat plants. This study provides information on structure elucidation of F-box proteins of a basidiomycetes plant pathogenic fungi and their role during pathogenesis.

The TRANSPORT INHIBITOR RESPONSE 1/AUXIN SIGNALING F-BOX (TIR1/AFB) protein serves as auxin receptor and links with Aux/IAA repressor protein leading to its degradation via SKP-Cullin-F box (SCFTIR1/AFB) complex in the auxin signaling pathway. Present study revealed 11 TIR1/AFB genes in wheat by genome-wide search using AFB HMM profile. Phylogenetic analysis clustered these genes in two classes. Several phytohormone, abiotic, and biotic stress responsive cis-elements were detected in promoter regions of TIR1/AFB genes. These genes were localized on homoeologous chromosome groups 2, 3, and 5 showing orthologous relation with other monocot plants. Most genes were interrupted by introns and the gene products were localized in cytoplasm, nucleus, and cell organelles. TaAFB3, TaAFB5, and TaAFB8 had nuclear localization signals. The evolutionary constraint suggested paralogous sister pairs and orthologous genes went through strong purifying selection process and are slowly evolving at protein level. Functional annotation revealed all TaAFB genes participated in auxin activated signaling pathway and SCF-mediated ubiquitination process. Furthermore, in silico expression study revealed their diverse expression profiles during various developmental stages in different tissues and organs as well as during biotic and abiotic stress. QRT-PCR based studies suggested distinct expression pattern of TIR1-1, TIR1-3, TaAFB1, TaAFB2, TaAFB3, TaAFB4, TaAFB5, TaAFB7, and TaAFB8 displaying maximum expression at 24 and 48 h post inoculation in both susceptible and resistant near isogenic wheat lines infected with leaf rust pathogen. Importantly, this also reflects coordinated responses in expression patterns of wheat TIR1/AFB genes during progression stages of leaf rust infection.