Transcriptome analysis revealed the potential mechanism of a wheat-Th. elongatum translocation line YNM158 against Fusarium head blight

Transcriptome analysis revealed the potential mechanism of a wheat-Th. elongatum translocation line YNM158 against Fusarium head blight | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Transcriptome analysis revealed the potential mechanism of a wheat-Th. elongatum translocation line YNM158 against Fusarium head blight Yi Dai, Wenlin Fei, Shiqiang Chen, Juntao Shi, Haigang Ma, Haifeng Li, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4079736/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Fusarium head blight (FHB) caused by Fusarium graminearum species complex is a destructive disease in wheat worldwide. Lack of FHB resistant germplasm is a barrier in wheat breeding for the resistance to FHB. Thinopyrum elongatum is an important relative species successfully used for genetic improvement in wheat. Results In this study, a translocation line YNM158 with a YM158 genetic background and carrying the fragment of diploid Th. elongatum 7EL chromosome created by 60 Co-γ radiation showed high resistance to FHB under both filed and greenhouse conditions. The transcriptome analysis validated that the horizontal transfer gene GST is one of the important contributors to FHB resistance in pathogen infection stage, whereas 7EL chromosome fragment also carries other genes regulated by F. graminearum during the colonization stage. In addition, the introgression of 7EL fragment affected the expression of wheat genes which were enriched in the resistance pathways including phosphatidylinositol signaling system, protein processing in endoplasmic reticulum, plant-pathogen interaction and MAPK signaling pathway at different stages after F. graminearium infection. Conclusions The study provides a novel germplasm for wheat resistance to FHB and new insights into the molecular mechanism of wheat resistance to FHB. Fusarium head blight wheat-Th. elongatum translocation line transcriptome analysis disease resistance pathway Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Introduction Fusarium head blight (FHB) is a primary disease caused by pathogens such as F. asiaticum and F. graminearium , seriously affecting yield and quality in wheat worldwide. Wheat infected with FHB, in addition to causing a significant reduction in yield, also accumulates the toxin deoxynivalenol (DON) in the seeds, which poses a significant risk to food safety. With global warming and changes in farming systems and practices, there is a trend of expanding the area of occurrence of the wheat blast, and the cultivation of blast-resistant varieties is the fundamental way to reduce its damage. However, the sources of resistance to wheat FHB were relatively narrow. There were only seven QTLs ( Fhb1 - Fhb8 ) related to FHB resistance, such as Fhb1 from chromosome 3BS and Fhb2 from chromosome 6BS of Sumai 3 [ 1 , 2 ], Fhb4 from chromosome 4B, Fhb5 from chromosome 5A Fhb8 from chromosome 7D of Wangshuibai [ 3 – 5 ], of which Fhb1 was recognized as the most stable master effect QTL for FHB resistance expansion and widely used in wheat breeding for the FHB resistance. Furthermore, the QTLs were also found in wheat relatives, such as Fhb3 from Leymus racemosus [ 6 ], Fhb6 from Elymus tsukushiensi [ 7 ] and Fhb7 from Thinopyrum ponticum [ 8 , 9 ]. As an essential close relative of wheat, tall wheatgrass has many important favorable traits, which represent a valuable source of alien gene resources for wheat. Studies have shown that there are three species of tall wheatgrass in nature, namely Th. elongatum (diploid, 2n = 2x = 14, E e E e ), Th. scirpeum (tetraploid, 2n = 4x = 28, E e E e E b E b ) and Th. ponticum (decaploid, 2n = 10x = 70, E e E e E b E b E x E x StStStSt). There is no scientific conclusion on the evolution process of polyploidy of the family in Thinopyrum , but it is believed that many interspecific hybridizations and natural doubling of chromosomes occurred during the evolution process of tall wheatgrass which was similar to common wheat [ 10 ]. Because tall wheatgrass has strong resistance to wheat FHB, so far, breeders have constructed wheat-tall wheatgrass chromosome addition line, substitution line and other genetic materials by distant hybridization using Th. elongatum and Th. ponticum , and obtained the results related to FHB resistance. For example, Jauhar et al. created 1E addition lines, 1E (1A) and 1E (1B) diploid substitution lines by crossing durum wheat with diploid Th. elongatum , and found that the 1E chromosome of diploid Th. elongatum may carry FHB resistance genes by FHB resistance evaluation [ 11 , 12 ]. Liu et al. (2017) obtained a disomic alien addition line with a pair of 7E Th. scirpeum chromosomes by hybridization of durum cultivar “Langdon” with the amphiploid 8801 (AABBEE) and found this addition lines showed high resistance to FHB [ 13 ]. Shen et al. identified the FHB resistance in the substitution lines 7E(7A), 7E(7B) and 7D(7E) from Th. elongatum , and 7el 2 from Th. ponticum in the Thatcher genetic background [ 14 ]. This FHB resistant locus, assigned as FhbLoP , was mapped to the distal region of the long arm of chromosome 7E in Th. ponticum within a 3.71 cM interval flanked by Xcfa2240 and Xswes19 , which accounts for 30.46% of the phenotypic variance [ 15 ]. This locus was then designated as Fhb7 and was fine mapped in 1.7 cM interval. Translocation lines with shortened Th. ponticum chromatin carrying Fhb7 was developed [ 8 ]. After that, Wang et al. [ 9 ] successfully cloned the Fhb7 gene from the Th. ponticum 7el 2 chromosome by assembling the genome of Th. elongatum . And they proved that this gene encoded a glutathione S-transferase (GST), which can open the epoxy group of DON toxin and catalyze its formation of glutathione adduct (DON-GSH), resulting in detoxification and anti-FHB effect. However, Guo et al. discovered that some wheat- Thinopyrum derivatives carrying the Fhb7 homologs had a different reaction to Fusarium head blight [ 16 ]. Similar results were also found in transgenic plants by overexpression of GST-encoding Fhb7 [ 17 ]. Moreover, because Th. elongatum, Th. scirpeum and Th. ponticum shares the E genome of Th. elongatum , so previous studies have shown that the FHB resistance genes are also located on the homologous group seven in diploid Th. elongatum [ 13 , 18 ]. For example, Zhang et al. incorporated a novel Fhb7 allele, Fhb7 The2 , into the wheat B genome through a small 7B-7E translocation (7BS·7BL-7EL) involving the terminal regions of the long arms, making this novel FHB resistance allele usable for breeding in both common and durum wheat [ 19 ]. It is well known that the introduction of chromosomes from wild species into wheat usually has linkage drag of undesirable genes and limits their application. In wheat genetic improvement, breeding translocation lines carrying alien beneficial genes, especially small fragment translocation lines, can reduce the linkage caused by alien chromosomes and has high genetic stability under common wheat genetic background, which is an ideal way to introduce alien beneficial genes. Therefore, in this study, we aim to: obtain the translocation lines with different size of 7EL chromosome fragment; develop the stable inheritance translocation line with FHB resistance; evaluate the application value of translocation lines in wheat breeding for FHB resistance; explore potential disease resistance genes and analyze the disease resistance pathways in translocation lines through transcriptome analysis. The results can not only provide new germplasm for wheat FHB resistance breeding, but also provide a theoretical basis for studying the resistance mechanism of wheat FHB. Result Establishment of a wheat- Th. elongatum 7EL chromosome translocation line YNM158 with FHB resistance When the alien chromosomes are introduced into wheat, there will be linkage drag, which will have a negative effect on the agronomic characters of wheat. So, creating the small segment translocation lines carrying the alien chromosomes can reduce the negative effects of linkage drag. In this study, the spikes of the translocation line T7BS·7EL, carrying the long arm of diploid Th. elongatum 7E chromosome, were treated by 60 Co-γ radiation during the flowering period of wheat. After treatment, the fresh pollen was collected and awarded to YM158 to obtain the M 1 seeds. The chromosomes of M 1 generation plants were identified by GISH, and the plants containing 7EL chromosome structure variation was selected for backcross with YM158. Finally, one line with stable agronomic characters was obtained in F 6 generation, which was named YNM158 (Fig. 1 A). The root-tip cells of YNM158 at mitotic metaphase were analyzed by GISH and FISH. Firstly, the presence of the translocation chromosome pair was confirmed in YNM158 by GISH (Fig. 1 B). Then, the chromosomal structural variation was observed on the end of 4BS chromosome according to the standard karyotype of CS (Fig. 1 C). Finally, the translocation chromosome can be represented as T7EL­4BS·4BL. In addition, the FHB resistance of YNM158 for two consecutive years showed that YNM158 has a high resistance to FHB in both field and greenhouse, and the level of FHB resistance was similar to that of SU3 (Fig. 1 D and Table 1 ). Moreover, we observed almost no difference for the tested agronomic traits, including spike length, number of grains per spike, number of spikelets and grain width, for 2 consecutive years (Fig. 1 E). However, plant height, thousand kernel, weight grain length and flag leaf area of YNM158 were significantly different compared with YM158 (Fig. 1 E). Table 1 Mean percentage of diseased spikelets (PDS) in different environments Lines 2020–2021 2021–2022 Field Greenhouse Field Greenhouse YNM158 4.81% ± 0.19d 5.18% ± 0.37c 5.21% ± 0.23d 5.27% ± 0.20d SU3 6.49% ± 1.79d 7.38% ± 2.89c 4.36% ± 0.22d 5.06% ± 0.21d AN8455 83.42% ± 10.36a 75.83% ± 14.69a 87.88% ± 5.51a 60.19% ± 6.24a YM23 28.79% ± 6.41c 53.69% ± 5.15b 15.54% ± 3.54c 21.87% ± 7.05c YM158 47.01% ± 11.92b - 31.80% ± 12.91b 45.09% ± 8.21b Note: The data were statistically analyzed by Kruskal-Wallis one-way ANOVA. Pairwise comparisons were completed using LSD. Different letters show significance at P < 0.05. RNA-seq data quality, assembly, and annotation of YNM158 and YM158 To analyze the genes associated with FHB resistance on chromosome 7EL chromosome segment in YNM158, RNA-sequencing (RNA-seq)-based transcriptome profiling was performed on the spikes which were inoculated F0609. In this experiment, total RNA was extracted from the spikes at 0-hours post-inoculation (hpi, no inoculation), 0.5 hpi, 2 hpi, 8 hpi, 24 hpi, 48 hpi, 72 hpi and 96 hpi, and 24 cDNA libraries were constructed (three repetitions per time). After filtering out the rRNAs and low-quality reads, a total of 193.57 GB high-quality clean data were obtained from 24 libraries (BioProject ID: PRJNA1011388), with an average of 80.65 GB clean data per library. The Q20 and Q30 were > 97% and > 93%, respectively. In addition, the GC contents were 48.86–52.53% among all samples. After assembly, the clean reads were mapped to the wheat and Th. elongatum reference genome (Chinese Spring v2.2 and Th. elongatum v1.0). On average, 91.17% of the reads were successfully aligned to the reference genome (Table S1 ). Therefore, these analyses indicated that the quality of our RNA-seq data was high and the sequencing depth was sufficient for further analysis. To investigate the impact of chromosome translocations on gene expression, RNA-seq-based transcriptome profiling was also performed on wheat variety YM158 which was one of the parents of YNM158. In this study, we constructed cDNA libraries (24 cDNA libraries) of YM158 after F. graminearum infection at different time. After filtering out the rRNAs and low-quality reads, a total of 188.56 GB high-quality clean data were obtained from 24 libraries (BioProject ID: PRJNA1011388), with an average of 78.57 GB clean data per library. The Q20 and Q30 were > 97% and > 94%, respectively. After assembly, the clean reads were mapped to the wheat reference genome (Chinese Spring v2.2). On average, 89.31% of the reads were successfully aligned to the reference genome (Table S1 ). Therefore, these analyses indicated that the quality of our RNA-seq data was high and the sequencing depth was sufficient for further analysis. Identification of the DEGs on 7EL chromosome post inoculation with F. graminearum By using the criteria of FDR 1, a total of 32102 differentially expressed genes (DEGs) were detected that significantly responses to F. graminearum infection at different times in YNM158, of which 222 DEGs were located on 7EL chromosome (Table S2 ). The 222 DEGs were analyzed, and the results showed that there were 60, 10, 25, 27, 49, 109 and 135 DEGs at 0.5 hpi, 2 hpi, 8 hpi, 24 hpi, 48 hpi, 72 hpi and 96 hpi, respectively (Fig. 2 A). GO function enrichment analysis indicated that these DEGs were enriched in catalytic activity, carboxylic acid transmembrane transporter activity and organic acid transmembrane transporter activity in terms of molecular function. And a total of 124 DEGs were enriched in catalytic activity (Fig. 2 B, Table S3). In terms of biological processes, organic anion transport, organonitrogen compound catabolic process and anion transport was the main role of the enriched genes. Among them, organonitrogen compound catabolic process was the term with the most enriched DEGs, with a total of 18 (Fig. 2 B, Table S3). And the top 10 GO terms with the lowest Q value were selected to draw the scatter diagram of enrichment items in Fig. 2 B. In addition, the KEGG pathway analysis revealed that the starch and sucrose metabolism pathway is the only one with the Q value less than 0.05 (Fig. 2 C, Table S4). Moreover, the heatmap of the DEGs enriched in this pathway showed the expression levels of these genes were down-regulated after infection with F. graminearum , especially in the first 8 hours after infection (Fig. 2 D). The top 10 pathways with the lowest Q value were selected to draw the scatter diagram of enrichment items in Fig. 2 C. Gene expression patterns analysis of DGEs on 7EL chromosome at different infection time-points The trend analysis of 222 DEGs on 7EL chromosome showed that the DEGs were clustered into 20 profiles, of which 153 DEGs were significantly clustered to the three profiles, including profile 19, profile 0 and profile 6, on the basis of P -value ˂ 0.05 (Fig. 3 A, Table S5). Among them, 77 genes showed a continuous upward trend with the extension of infection time (profile 19), and 56 genes showed a continuous downward trend (profile 0). In addition, the 222 DEGs were analyzed of the weighted gene co-expression network analysis (WGCNA) modules associated with infection time. The selection of a soft threshold (Power) is the key step to constructing the network. When the soft threshold was set as 10 with a scale-free network fitting index of R 2 > 0.80, average connectivity was close to 0 (Fig. S1 A). The hierarchical cluster tree was drawn based on the optimal soft threshold, and the genes clustered in the same branch are divided into the same module. Finally, four modules, including turquoise, blue, brown and grey, were obtained (Fig. S1 B). Among them, the brown module was significantly correlated with infection at 0.5 hpi (R = 0.92, P ≤ 0.05), the turquoise module was significantly correlated with infection at 8 hpi (R = 0.82, P ≤ 0.05), and the blue module was significantly correlated with infection at 96 hpi (R = 0.69, P ≤ 0.05). Based the cut-off criteria |module member-ship| (|MM|) > 0.8 and |gene significance| (|GS|) > 0.8, 19 genes with high connectivity in the clinically significant module were identified as hub genes (Fig. 3 B and 3 C, Table S6). The DEGs and hub genes obtained from trend analysis and WGCNA, respectively, were sued to draw the Venn diagrams, and a total of 12 genes were obtained in both analyses (Fig. 3 D, Table 2 ). The qRT-PCR verification of 12 DEGs showed that the relative expression levels of several genes were induced by F. graminearium , such as Tel7E01G1020600 , Tel7E01G943900 and Tel7E01G980900 . Among them, Tel7E01G1020600 encoded glutathione S-transferase (GST), whose expression was induced by F. graminearium and significantly upregulated from 72 hpi (Fig. 3 E). Tel7E01G943900 and Tel7E01G1980900 had similar expression patterns, both of which were up-regulated at the early stage of infection. Tel7E01G943900 encodes a receptor-like kinase, and the expression of this gene was up-regulated by about 5 times at 0.5 hpi, but then began to be down-regulated (Fig. 3 F). Similarly, Tel7E01G1980900 encoded a monosaccharide-sensing protein, the expression of this gene was significantly reached the highest level at 2 hpi, which was about 4 times than that at 0 hpi, but then began to be down-regulated and was almost no expression after 24 h of infection (Fig. 3 G). These results suggested that there may be multiple resistance genes on chromosome 7EL fragment, which provide resistance to FHB at different stages of F. graminearium infection. Table 2 The gene was verified by qRT-PCR. Gene ID Gene description Tel7E01G1002700 Lysine ketoglutarate reductase trans-splicing-like protein (DUF707) Tel7E01G1013700 Galactoside 2-alpha-L-fucosyltransferase Tel7E01G1020600 Glutathione S-transferase Tel7E01G211400 Protein kinase Tel7E01G899900 NF-X1-type zinc finger protein NFXL1 Tel7E01G905000 Disease resistance protein (NBS-LRR class) family Tel7E01G934300 Carbonic anhydrase Tel7E01G939300 Receptor-like kinase Tel7E01G941500 Carboxypeptidase Tel7E01G943900 Receptor-like kinase Tel7E01G946300 Blue copper binding protein Tel7E01G980900 Monosaccharide-sensing protein 2 Extraction of wheat DEGs between YM158 and YNM158 after F. graminearum infection Since wheat FHB is a compatible disease, Stephens et al., (2008) divided the infection process into initial colonization stage, infection stage and late infection stage [ 20 ]. In this study, we analyzed the effect of the introduction of 7EL chromosome fragments on FHB resistance using 8 hpi as the cut-off point. And we defined the initial colonization stage before 8hpi, the infection stage after 8hpi. First of all, 12761 and 15719 DEGs were obtained in the initial colonization stage and the infection stage, respectively (Table S7 and S8). Then, the DEGs at 0 hpi between YM158 and YNM158 were removed (Table S9). Finally, a total of 4734 DEGs were obtained at the initial stage of colonization (Fig. 4 A), and 10489 DEGs were obtained at the stage of infection (Fig. 4 B). After alignment with the reference genome, 4060 and 9808 wheat DEGs were screened for subsequent analysis at the initial colonization stage and infection stage, respectively (Fig. 4 C, Table S10). KEGG pathway enrichment analysis of wheat DEGs at different stages To analyze the effect of 7EL chromosome on the resistance pathway of wheat at different stages after F. graminearum infection, the wheat DEGs were subjected to KEGG pathway enrichment analysis. We identified 7 pathways that were significantly enriched with a Q value less than 0.05 during the initial colonization stage, among which phosphatidylinositol signaling system was the most enriched pathway with the lowest Q value and the protein processing in endoplasmic reticulum pathway was the most enriched pathway with the DEGs number (Fig. 5 A, Table S11). Further analysis of the genes involved in these pathways was found that the expression of some genes related to phosphatidylinositol 4-phosphate 5-kinase (PIP5K), immunoglobulin-binding protein (BIP4) and heat shock protein (Hsp) in YNM158 was higher than that in YM158 after F. graminearum infection (Fig. 5 B). At the infection stage, 21 specific pathways were significantly enriched with a Q value less than 0.05. Among them, glutathione metabolism was the pathway with the lowest Q value and the largest number of DEGs (Fig. 5 C, Table S11). Moreover, genes related to ascorbate peroxidase (APX), glutathione reductase (GR), glutathione-S-transferase (GST) in this pathway were up-regulated in YNM158 (Fig. 5 D). In addition, we also found that the genes related to ABC transporter (ATP-binding cassette, ABC) were up-regulated in YNM158, they may be associated with the transportation of DON (Fig. 5 D). The plant-pathogen interaction pathway was the second most enriched pathway, with a total of 152 DEGs enriched in this pathway (Fig. 5 C). Through the heat map of gene expression, we found that the expression levels of some genes related to hypersensitive response (HR), such as TraesCS2D03G0030700 and TraesCS2D03G1070500 , began to be up-regulated at 24 hpi with F. graminearum in YNM158 (Fig. 5 D). It is well known that secondary metabolites play an important role in plant resistance to pathogens. In our study, a total of 13 pathways were identified in both initial colonization stage and infection stages with a Q value less than 0.05. Among them, biosynthesis of secondary metabolites was the pathway with the lowest Q value (Fig. 5 E and 5 F, Table S12). The analysis of gene expression in this pathway found that some genes related to flavonol synthase (FLS), chalcone isomerase (CHI), chalcone synthase (CHS) and hydroxycinnamoyl-CoA shikimate (HCT) showed a significantly up-regulated expression trend with the extension of F. graminearum infection time in YNM158 (Fig. 5 G). In addition, in the MAPK pathway, we also found the expression of some genes related to respiratory burst oxidase (RBOH) was significantly up-regulated after F. graminearum infection, and the expression level was the highest at 72 hpi, while the expression of some genes related to mitogen-activated protein kinase (MAPK) was up-regulated at initial colonization stage and then down-regulated at the infection stage in YNM158 after F. graminearum infection in YNM158 (Fig. 5 G). WGCNA of wheat DEGs The 12661 wheat DEGs were also analyzed of the WGCNA modules (Table S13). The selection of a soft threshold (Power) is the key step to constructing the network. When the soft threshold was set as 9 with a scale-free network fitting index of R 2 > 0.80, average connectivity was close to 0 (Fig. 6 A and 6 B). The hierarchical cluster tree was drawn based on the optimal soft threshold, and the genes clustered in the same branch are divided into the same module (Fig. 6 C). Finally, 14 modules, which were correlated with varieties were obtained. Among them, the yellow module was significantly positive associated with YNM158 (R = 0.96, P ≤ 0.05), and a total of 847 DEGs were obtained in this module (Fig. 6 D, Table S14). Screened the core genes at different infection stages The Venn diagrams was drawn between the wheat DEGs at different infection stages and the hub genes positive associated with YNM158. At initial colonization stage, a total of 120 DEGs were obtained from the specific enrichment pathways (Table S15), and among them, 13 DEGs may be associated with the FHB resistance of YNM158 (Fig. 7 A, Table S16). At infection stage, 830 DEGs were obtained from the specific pathway (Table S17), of which 10 DEGs may be related to the FHB resistance of YNM158 (Fig. 7 B, Table S16). In addition, among the same pathways in both stages mentioned above, we obtained 202 DEGs (Fig. 7 C, Table S18), 21 of which may be related the FHB resistance of YNM158 (Fig. 7 D, Table S16). In order to verify the correlation between the core genes and the stages of FHB resistance, these selected genes were verified by RT-PCR. Compared with the expression of YM158 at 0 hpi, we found that the expression levels of 6 wheat genes in YNM158 were significantly up-regulated after F. graminearum infection (Table 3 ). Among them, TraesCS4D03G0528700 and TraesCS4B03G0573000 belong to the phosphatidylinositol signaling system and protein processing in endoplasmic reticulum pathway, respectively. And had the same expression pattern at initial colonization stage, both of which were significantly up-regulated in YNM158. TraesCS4D03G0528700 had the highest expression level at 2 hpi (Fig. 7 E) and the expression of TraesCS4B03G0573000 reached the highest level at 8 hpi (Fig. 7 F). TraesCS2D03G0030700 and TraesCS7D03G0466200 encoded NBS-LRR disease resistance protein and 3-ketoacyl-CoA synthase, respectively, both belong to the plant-pathogen interaction pathway. However, their expression patterns were slightly different. The expression of TraesCS2D03G0030700 was significantly up-regulated after F. graminearum infection and the expression of this gene in YNM158 was always higher than that of YM158 (Fig. 7 G). Although TraesCS7D03G0466200 was also induced after F. graminearum infection, the expression of TraesCS7D03G0466200 in YNM158 was significantly higher than that in YM158 until 48 hpi (Fig. 7 H). In addition, we found that the expression of one gene in biosynthesis of secondary metabolites and MAPK signaling pathway-plant was induced by F. graminearum . The results of RT-PCR indicated that these two genes may mediate the resistance of YNM158 to FHB during the initial colonization stage and infection stage. TraesCS7A03G1308100 encoded hydroxycinnamoyl-CoA shikimate, the expression of this gene in YNM158 was significantly higher than that in YM158 from 8 hours after F. graminearum infection (Fig. 7 I). On the contrary, the expression of TraesCS1A03G0718100 was significantly up-regulated at 0.5 hpi in YNM158. However, there was little difference in the expression of this gene between YM158 and YNM158 after 48 hpi (Fig. 7 J). The above results suggest that the introduction of 7EL chromosome fragments may affect the disease resistance pathway of wheat, thereby improving the FHB resistance of wheat. Table 3 The core genes verified by RT-PCR at different infection stages. Gene ID Pathway Gene description Initial colonization stage TraesCS4D03G0528700 Phosphatidylinositol signaling system Phosphatidylinositol-4-phosphate 5-kinase family protein TraesCS4B03G0573000 Protein processing in endoplasmic reticulum 70 kDa heat shock protein Infection stage TraesCS2D03G0030700 Plant-pathogen interaction NBS-LRR disease resistance protein TraesCS7D03G0466200 Plant-pathogen interaction 3-ketoacyl-CoA synthase Both stage TraesCS7A03G1308100 Biosynthesis of secondary metabolites Hydroxycinnamoyl-CoA TraesCS1A03G0718100 MAPK signaling pathway - plant Respiratory burst oxidase-like protein Discussion YNM158 can be effectively applied in FHB improvement in wheat breeding Breeding and applying FHB resistant varieties in wheat production is an effective way to control the destructive disease. However, the long-term intraspecific cross breeding of wheat reduced the range of genetic variation among varieties and had poor resistance to FHB, while the related wild species and genera of wheat carry many FHB resistance genes. For example, the FHB resistance genes Fhb3 , Fhb6 and Fhb7 on chromosome 7Lr#1S of Leymus racemosus , chromosome 1E(ts)#1S of Elymus tsukushiensis and chromosome 7el 2 of Th. ponticum , respectively were reported to have major resistance to FHB [ 6 – 9 ]. In addition, the 1Y c and 3S c chromosomes of Roegneria ciliaris [ 21 ], the 1E, 7E chromosome of diploid Th. elongatum , the chromosome 3St of Elymus repens [ 22 ], and the 7M g chromosome of Aegilops geniculata [ 23 ] also possessed the FHB resistance genes. Although the introduction of alien chromosomes can improve the resistance to FHB, it also brings some genetic encumbrance, which makes the agronomic characters of most foreign germplasm poor and difficultly to be directly used in wheat breeding for FHB resistance. Therefore, to make full use of wheat related species in wheat breeding for FHB resistance, it is necessary to create small fragment translocation lines to develop new varieties with increased FHB resistance and no yield penalty. At present, Fhb7 gene from Th. ponticum was poured into cultivated wheat through small segment translocation lines and used in wheat breeding for FHB resistance [ 9 ]. The diploid form of tall wheatgrass, Th. elongatum , has a high level of resistance to FHB and was used to increase FHB resistance in wheat cultivar Chinese Spring by translocation development [ 19 , 24 ]. In this study, we have successfully established the translocation lines with the small fragments of chromosome 7EL from diploid Th. elongatum by physical radiation, one of which has excellent agronomic traits and high resistance to wheat FHB and named YNM158. Different from previous reports, the translocation in YNM158 occurred on chromosome 4BS, a non-compensative translocation, and the reason for this phenomenon was that chromosome translocation was induced by ionizing radiation, the breakage and reconnection of wheat and alien chromosomes were random, so most of them were uncompensated translocations. Interestingly, the survey results of agronomic traits for two consecutive years showed that YNM158 did not have any bad performances due to a non-compensatory translocation line, such as genetic instability of exogenous chromosomes, high plant height and poor fertility. This may be because a small alien chromosome fragment was attached to the distal end of 4BS chromosome in recurrent parent in YNM158, whereas no chromosome fragments were lost in wheat. And previous studies have also shown that non-compensatory translocation lines also have important utility in wheat breeding. For example, the small fragment translocation line 5VS-6AS·6AL can be used to improve the quality of wheat soft grains [ 25 ], the 3A-7J s translocation line can be used to improve wheat stem rust resistance [ 26 ], two homozygous translocation lines T1AS·1AL-6VS and T4BS·4BL-6VS-4BL carrying Pm21 can be used to enhance powdery mildew resistance in wheat [ 27 ]. Therefore, we propose the translocation line YNM158, contained small fragment of 7EL chromosomes, has a good application prospect in the breeding for resistance to FHB in wheat. Transcriptome analysis validated that glutathione metabolic is one of the important contributors to FHB resistance in pathogen infection stage Transcriptome analysis is a valuable tool in investigating the molecular mechanisms behind cereal resistance to fungal infections as the costs of this technique decrease and its applications become more widespread. In this study, we found that compared with YM158, the glutathione metabolic was the most significantly enriched pathway in YNM158 after F. graminearum infection during the infection stage (Fig. 5 D). It is well known that plants respond to fungal infections by activating defense genes including producing reactive oxygen species (ROS) which can enhance the strength of plant cell wall to resist the invasion and colonization of pathogens [ 28 , 29 ]. However, when ROS accumulates in large amounts in plants, it will cause oxidative stress, damage plant cells, and lead to cell dysfunction and even death. Currently, the enzymes involved in the antioxidant defense system can be divided into two groups: (i) Enzymatic antioxidants such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase GPX, glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), and dehydroascorbate reductase (DHAR); (ii) Non-enzymatic antioxidants such as ascorbic acid (AA), reduced glutathione (GSH), α-tocopherol, carotenoids, plastoquinone/ubiquinone and flavonoids [ 30 ]. Early research reported that ascorbate-glutathione (AsA-GSH) cycle is the important way to diameter of removal of ROS in plant. Among them, APX is the key enzymes of this cycle, which can utilize AsA as the electron donor reducing H 2 O 2 to water, and prevents the accumulation of a toxic level of H 2 O 2 in photosynthetic organisms under stress conditions [ 31 ]. And the glutathione (GSH) participate in various metabolic processes and were the essential components of antioxidative and detoxification systems in plant cells [ 32 ]. It can be used as both a reducing agent and a strong nucleophile, participating in the elimination of reactive oxygen species (ROS) through thiol-disulphide redox reactions, and in the detoxification of various heterogenic organisms through conjugation reactions, respectively [ 33 ]. However, GSH will be oxidized to oxidized glutathione (GSSG) in the reaction of clearing ROS. In order to maintain the balance of GSH content in the plant, GR enzyme will effectively and timely reduce GSSG to GSH. It can be seen that GR enzyme plays a very important role in clearing ROS and maintaining the content of GSH in plants. For example, overexpression of GR gene from Haynaldia villosa in wheat can increase resistance to powdery mildew [ 34 ]. In this study, we found that the expression of some APX-encoding and GR-encoding genes in YNM158 was up-regulated during the infection stage (Fig. 5 D). Therefore, we suggest that, glutathione may play a key role in ROS-mediated resistance to FHB in wheat. In addition, glutathione S-transferase (GST) represents a group of multifunctional enzymes widely present in plants and plays important roles in plant secondary metabolism [ 35 ], growth and development [ 36 ], and biotic and abiotic stress responses [ 37 ]. Furthermore, one of its most important functions is the ability to inactivate toxic compounds. Because GST can form complexes with glutathione (GSH) by catalyzing hormones, toxins to inactivate or eliminate toxicity of many substances, and expel them in the body under the action of relevant transporters [ 38 ]. These results suggested that GST played a crucial role in plant disease resistance. For example, NbGSTU1 can increase the resistance to Colletotrichum destructivum in Nicotiana benthamiana [ 39 ]. The lack of GSTU13 function resulted in enhanced disease susceptibility toward several fungal pathogens in Arabidopsis thaliana [ 40 ]. Overexpression of LrGST5 in tobacco can improve the resistance of transgenic plants to F. oxysporum [ 41 ]. TaGSTU6 interactions can enhance wheat resistance to powdery mildew but not wheat stripe rust [ 42 ]. We know that wheat infected with FHB can be contaminated with a variety of mycotoxins, especially deoxynivalenol (DON) [ 43 ]. It has been reported that GSH can form a GSH-DON conjugates under the catalysis of GST to reduce the accumulation of DON and protect plants from toxicity. For instance, Fhb7 and FhbRc1 , encoding glutathione S-transferase, enhanced the resistance to FHB in wheat background [ 9 , 44 ]. In this study, the expression of GST- encoding genes, including TraesCS1A03G0109100 , TraesCS3D03G0946300 , TraesCS4D03G0493500 , TraesCS5B03G0050700 , TraesCS5A03G0730500 , TraesCS5B03G0770700 and Tel7E01G1020600 was significantly up-regulated after infection with F. graminearum in YNM158. Among them, the expression of the Fhb7 homolog Tel7E01G1020600 increased sharply at 72 hpi, which was tens of times higher than that of the non-infection (Fig. 3 E). It can be seen that GST is one of the important contributors to FHB resistance roles in pathogen infection stage. However, the Tel7E01G1020600 in YNM158 was derived from diploid Th. elongatum , which is not consistent with the origin of Fhb7. So, whether Tel7E01G1020600 in YNM158 has the same disease resistance function as Fhb7 needs to be further studied. Other genes from 7EL fragment in YNM158 might also be involved in increasing FHB resistance especially in pathogen initial colonization stage DON toxin is a very important fungal pathogen when F. graminearum infects wheat. It can synthesize a large amount of F. graminearum along the inflorescence axis and promote the process of disease expansion. However, some studies have reported that when the pathogen initially infected wheat anthers, there was no DON synthesis signal, and only when the disease spread along the inflorescence axis from the inoculation point, DON began to be synthesized in large quantity in the pathogen [ 45 , 46 ]. It can be seen that DON can help the pathogen spread along the wheat spike axis, but it is not necessary for its initial infection [ 47 ]. In the process of long-term co-evolution between plants and pathogens, a series of complex defense mechanisms have gradually formed. Generally, pathogen-associated molecular pattern (PAMP)-trigged immunity (PTI) is the first defensive line of plant innate immunity and is mediated by pattern recognition receptors (PRRs). And the PRRs are divided into two types, receptor-like kinases (PLKs) and receptor-like proteins (PLPs). To date, many PLKs have been found to play a key role in wheat disease resistance. For example, Sun et al. (2023) reported that a repeat receptor-like kinase-encoding gene TaBIR1 contributed to wheat resistance against Puccinia striiformis f. sp. tritici by mediating ROS production and callose deposition [ 48 ], and the cysteine-rich receptor-like kinase TaCRK3 contributed to defense against Rhizoctonia cerealis in wheat through directing antifungal activity and heightening the expression of defense-associated genes in the ethylene signaling pathway [ 49 ]. And the RLKs have also been found to contribute to grain resistance to Fusarium resistance in cereals. For instance, Thapa et al. (2018) identified two homologous genes on barley chromosome 6H (HvLRRK-6H) and wheat chromosome 6DL (TaLRRK-6D), respectively, which could enhance cereal resistance to FHB disease [ 50 ]. And the Arabidopsis senses Fusarium elicitors in early immune responses to extracts from Fusarium spp. by a novel receptor complex which was encoded by the leucine-rich repeat receptor-like kinase MDIS1-interacting receptor-like kinase 2 (MIK2) at the cell surface [ 51 ]. Interesting, we also identified several RLKs-encoding genes on the 7EL fragment in this study, and the expression of them was significantly up-regulated at initial colonization stage after F. graminearum inoculation, such as the expression of Tel7E01G943900 which was significantly up-regulated at 0.5 hpi (Fig. 3 F). In addition, in immune responses, plants have developed a number of disease-resistance mechanisms to resist nutrient uptake by pathogens, which involve sugar transport, metabolism, and signal transduction. The previous studies have shown that hexose released by cell wall invertase (CWIN) activity not only acts as a signal molecule to trigger the expression of disease-resistance related genes, but also is an essential metabolite and energy source for the synthesis of antioxidant compounds and defense molecules, such as salicylic acid and callose [ 52 – 54 ]. For example, AtSTP4 and Atβfruct1 encoding monosaccharide transporter and CWIN, respectively, are both induced in Arabidopsis during parasitic infection by fungus [ 55 ]. Chang et al. (2020) reported that silence the hexes transporter-encoding gene PsHXT1 in wheat stripe rust can significantly inhibit the pathogenicity of pathogenic bacteria [ 56 ]. In this study, the expression of a monosaccharide-sensing protein-encoding gene Tel7E01G980900 was significantly up-regulated within 8h after infection with F. graminearum and reached the highest level at 2 hpi in YNM158 (Fig. 3 G). And knowledge of the function of monosaccharide-sensing protein is similar to that of hexose transporters [ 57 , 58 ]. Therefore, we speculated that they can also be conducted with CWINs to bring hexose back to host cells, reducing sugar availability to the pathogen, and thus improve host disease resistance. But this needs to be confirmed in the further studies. It is well known that F. graminearum is a kind of facultative trophic bacteria. Therefore, wheat needs to use a series of defense mechanisms to resist pathogen infection at different stages. The introduction of 7EL chromosome fragments not only brought GST-encoding gene which was one of the important contributors on DON detoxification, but also brought other genes which were up-regulated at initial colonization stage. And these genes were also involved in increasing FHB resistance. Therefore, in-depth study of these genes can provide new insights into the molecular mechanisms of wheat resistance to FHB. Introgression of 7EL fragment altered the gene expression in wheat after F. graminearum inoculation The introduction of alien chromosome fragments not only brought the resistance genes, but also affects gene expression on normal chromosomes [ 59 , 60 ]. In this study, we also found that the translocation of chromosomes affected the expression of wheat genes which were enriched in the resistance pathways including phosphatidylinositol signaling system, protein processing in endoplasmic reticulum, plant-pathogen interaction and MAPK signaling pathway at different stages with F. graminearium infection. When plants are infected with pathogens, phospholipase C (PLC) is rapidly activated by different pathogen-associated molecular patterns (PAMPs) and effector proteins in plant cells [ 61 ]. And then catalyze phosphatidylinositol 4-phosphate (PI4P) and phosphatidylinositol (4,5) bisphosphate [PI(4,5)P2] to produce inositol 2-phosphate (IP2) or inositol 3-phosphate (IP3) and diacylglycerol (DAG). These are conserved compounds of pathogenic microbes that are perceived by immune receptors present in resistant plants [ 62 , 63 ]. The previous studies reported that silencing and knock‑out SlPLC2 in tomato can reduce susceptibility to Botrytis cinereal [ 61 , 62 ]. And the SlPLC6 plays a key role in both for Ve1 resistance protein mediated resistance to Verticillium dahliae and Pto/Prf resistance protein mediated resistance to Pseudomonas syringae [ 64 ]. In addition, it has been reported that salicylic acid (SA), jasmonate (JA)and methyl jasmonate can increase the expression of OsPI-PLC in rice ( Oryza sativa ) and improve the resistance of rice to Magnaporthe oryzae [ 65 ]. In this study, it was found that the expression of some PLC genes in YNM158 was higher than that in YN158 at the initial colonization stage, such as TraesCS4A03G0225500 and TraesCS4B03G0547100 . Moreover, the results of qRT-PCR showed that the expression of TraesCS4D03G0528700 , which encoded phosphatidylinositol 4-phosphate-5 kinase (PIPK5), in YNM158 was higher than that in YM158 at initial colonization stage (Fig. 7 B). We know that PIP5K was the catalytic enzyme for the synthesis of PI(4,5)P2 and Shimada et al., (2019) have pointed out that the biosynthesis of PI(4,5)P2 was an important target to improve the defense ability of Arabidopsis thaliana against Colletotrichum, and its activity also determines the defense ability of Arabidopsis thaliana against Colletotrichum [ 66 ]. Therefore, we speculated that PIP5K gene can affect the accumulation of PI(4,5)P2 in YNM1158 to participate in the PLC-mediated response to F. graminearium infection, and thus affect the colonization of F. graminearium to improve the resistance to FHB during the initial stages of infection. There are also defense-related proteins in plants that are synthesized by the rough endoplasmic reticulum (RER), so when plants attacked by the pathogen, the genes encoding endoplasmic reticulum (ER) chaperones are induced, such as the immunoglobulin-binding protein (BIP), heat shock protein (Hsp), calreticulin (CRT) and protein disulfide isomerase (PDI)-encoding genes. The previous studies have shown that the Hsp was one of the ER chaperones, which play an indispensable role as molecular chaperones in the quality control of PRRs and intracellular resistance (R) proteins against potential invaders [ 67 ]. For example, Hsp90 was not only involved in the defense of many microbial pathogens by activating the cytosolic R proteins containing nucleotide-binding domain and a leucine-rich repeat, but also participated in chitin responses and anti-fungal immunity in a chaperone complex with its co-chaperone Hop/Sti1 [ 67 , 68 ]. In terms of specific diseases, cytoplasmic Capsicum annuum Hsp70 (CaHsp70) can enhance the resistance to Xanthomonas campestris pv. vesicatoria in pepper [ 69 ], GmHsp40 can increase soybean resistance to Soybean mosaic virus [ 70 ], Hsp70 can enhance the resistance to powdery mildew in cucumber under heat shock-induction [ 71 ], MeHsp90.9-MeSGT1-MeRAR1 chaperone complex interacted with MeATGs to trigger autophagy signaling to improve disease resistance to cassava bacterial blight [ 72 ]. In this study, we found the expression of TraesCS4B03G0573000 , which encodes the heat shock protein in YNM158 significantly induced to be upregulated after infection by F. graminearium at initial colonization stage, which was opposite to the expression pattern in YM158 (Fig. 7 F). Usually, the expression of the genes encoding ER chaperones even predates the expression of genes encoding pathogenesis-related (PR) proteins [ 73 ]. Therefore, we inferred that the expression of genes encoding Hsp protein would rapidly induce after infection with F. graminearium in YNM158, thus activating the defense mechanism earlier, inducing programmed cell death, affecting the colonization of pathogens, and making plants resistant to disease, which provides a new idea for further research on the mechanism of FHB resistance. Reactive oxygen species (ROS) are the important signaling molecules in defense responses during plant-pathogen interactions, which are mainly produced by respiratory burst oxidase homologs (RBOHs) [ 74 ]. In Arabidopsis, AtRBOHD and AtRBOHF are responsible for ROS production against pathogen attacks [ 75 ]. In Nicotiana benthamiana , NbRBOHA and NbRBOHB silencing leads to less ROS production and reduced the resistance against the infection by potato pathogen Phytophthora infestans [ 76 ]. Phosphorylation is known to be one of the essential mechanisms of RBOHD activation and is also transcriptionally activated by some kinases, such as MAPK cascades, and the transcriptional regulation of RBOHs may play a key roles in subsequent ROS bursts after turnover of the plasma membrane-localized RBOHs used for the first burst [ 77 ]. For example, Yamamizo et al. (2007) reported that MAPK kinase was involved in inducing the response of potato StRBOHC and StRBOHD genes in response to pathogen signals in potato [ 78 ], and Asai et al. (2008) also illustrated that the MAPK cascade MEK2-SIPK regulates the oxidative burst resulting from the induction of RBOHB expression in resistance to P. infestans and Colletotrichum orbiculare of N. benthamiana [ 79 ]. Here, we identified the expression of a RBHO-encoded gene TraesCS1A03G0718100 was up-regulated after F. graminearum infection in YNM158 (Fig. 7 J), as well as some genes encoding MAPK kinase (Fig. 5 G). Therefore, we hypothesized that MAPK kinase in YNM158 might be involved in inducing RBOH gene response to the resistance against F. graminearum . However, this puzzle requires further investigations. Conclusions FHB is a devastating wheat disease that seriously affects the yield and quality of wheat. Up to now, many laboratories around the world have carried out researches on wheat resistance to FHB. It has been proved by practice that the most economical and effective way to resist the damage of wheat FHB is to mine the genes with high resistance to FHB and breed new varieties resistant to FHB. In this study, a translocation line YNM158 carried 7EL chromosome fragment obtained by distant hybridization not only has excellent resistance to FHB, but also had stable agronomic traits that could potentially be used in FHB resistance breeding. Moreover, transcriptome analysis showed that the 7EL chromosome fragment not only carried the genes that can detoxify DON, but also have the genes that can affect the colonization of F. graminearum during the early stage of infection. In addition, the introgression of 7EL fragment altered the gene expression and activated the special resistance pathway in YNM158 after F. graminearum inoculation. It is demonstrated that YNM158 may have a variety of molecular mechanisms to against F. graminearum infection, and show a high resistance to FHB phenotype. Therefore, these results not only provide a new germplasm for wheat resistance to FHB, but also provide new insights into the molecular mechanism of wheat resistance to FHB, and also provide a new way for breeding new varieties with high resistance to FHB. Materials and Methods Plant materials Triticum aestivum cv. Chinese Spring (CS), Su Mai3 (SU3), An Nong8455 (AN8455), and Yang Mai158 (YM158) are maintained at Yangzhou University, China. Wheat cultivar YM158 was pollinated with the 60 Co-γ-irradiated pollen of the long-arm translocation line TW-7EL1 (T7BS·7EL) of 7E chromosome with excellent FHB resistance developed in previous studies. The small fragment of 7EL chromosome translocation line Yangnongmai158 (YNM158) was selected from the hybrid progeny in this study. Cell cycle synchronization and preparation of mitotic chromosomes Cell cycle synchronization and slide preparation followed Lei et al. [ 80 ] with minor modifications. Seeds were soaked in water for 3–5 hours and germinated on moist filter paper for 2 days in the dark at 25°C. When the roots grew to about 2.5 cm long, the roots were treated with 2 µmol/L amiprophosmethyl (APM) for 2.5 h. Then the root tips were cut off and treated in a nitrous oxide gas chamber for 1 h. After that the root tips were fixed in ice-cold 90% acetic acid for 8 min, washed with sterile distilled water (ddH 2 O) and stored in 70% ethanol at -20°C until use. For slide preparation, root tips were washed in ddH 2 O for 5 min. The apical meristem of roots was cut and incubated in 25 µL of enzyme solution containing 2% cellulase Onozuka R-10 (Yakult Pharmaceutical, Tokyo) and 1% pectolyase Y23 (ICN) for 1 h at 37°C in a water bath. Meristems were separated with a needle in 50 µL of 100% acetic acid and immediately dropped onto microscope slides using a pipette at a height of approximately 10 cm and then placed in a wet box for about 20 min. The number and location of chromosomes were observed and recorded under the phase contrast objective (Nikon 80i, Japan), and the well-prepared slides were stored in a -70°C refrigerator until use. Genomic in situ hybridization (GISH) and Fluorescence in situ hybridization (FISH) analysis The techniques of GISH and ND-FISH followed those of Tang et al. [ 81 ]. Total genomic DNA of Th. elongatum was labeled with digoxigenin-12-dUTP by Nick Translation method and used as a probe for GISH. The repetitive sequences oligo-pSc119.2 and oligo-pAs1 were synthesized as the probes. And the 5’ end of oligo-pSc119.2 was labeled by 6-carboxyfluorescein (6-FAM), the 5’ ends of oligo-pAs1 was labeled by 6-carboxytetramethylrhodamine (Tamra). The labeled probes were dissolved in 2 × SSC and 1× TE buffer (pH 7.0), dropped to the prepared slides. After that the slides were covered with a coverslip and placed in a humidified hybridization cassette at 37°C for 10 h, and then transferred into 2×SSC for 2 min at room temperature. Finally, the slides were quickly dried, and then 6.5 µL DAPI was added to each slide (Vector, No. H-1200). After ND-FISH analysis, the slides were washed in 2×SSC for 2 min at room temperature. After drying the same slides were subjected to GISH analysis. Hybridization signals were observed using a fluorescent microscope and images were obtained with a CCD camera (Color Cooled Digital DS-Fi1c, Nikon 80i, Japan). Evaluation of Disease Resistance The translocation line YNM158 and its hybrid offspring were screened for FHB resistance in the field and greenhouse in 2021 and 2022. At early flowering stage, the central spikelet was injected into 10 µL fungal suspension (50,000 spores/mL), and at least three spikes from each plant was injected. Following inoculation, the plants were misted for 72 h for FHB development. After 21 days, all infected spikelets per inoculated spike were counted. Wheat cultivars An8455 served as the susceptible controls in both the field and greenhouse, respectively, and SU3 served as the resistant control in both the field and greenhouse. De novo assembly of RNA-seq reads and quantifying gene expression The transcriptome analysis was comprised of 8 time points after inoculation with F. graminearum : 0 hpi, 0.5 hpi, 2 hpi, 8 hpi, 24 hpi, 48 hpi, 72 hpi and 96 hpi. Three spikes after inoculation was randomly selected for each time were mixed to extract RNA. Total RNA was extracted using Trizol reagent kit (Invitrogen) according to the manufacturer’s protocol. RNA quality was assessed on an Agilent 2100 Bioanalyzer (Agilent Technologies) and checked by using RNase-free agarose gel electrophoresis. The mRNA was enriched by Oligo (dT) beads. Then the enriched mRNA was fragmented and used as the template for cDNA synthesis. The cDNA fragments were sequenced using Illumina HiSeq2500 by Gene Denovo Biotechnology Co. For the analysis process after sequencing, referred to the article published by Dai et al. [ 82 ]. The genome of Chinese Spring (IWGSC RefSeq v2.1) and the genome of Th. elongatum (GWHABKY00000000) were used as the reference genome in this study. The obtained sequences were submitted on the sequence read archive data (BioProject ID: PRJNA1011388). Quantitative real-time polymerase chain reaction Three spikes after inoculation was randomly selected for each time were mixed to extract RNA. qRT-PCR was performed under the following program: 94°C for 5 min, and then 40 cycles: 94°C for 5 min followed by 60°C for 30 s. For the melt curve analysis, the following program was included after 40 cycles: 95°C for 10 s followed by 60°C for 30 s and a constant increase from 60 to 95°C. The relative expression levels were determined using the ­2 −ΔΔCt method. qRT-PCR assays were carried out in three independent biological samples pretreatment and three technical replicates per samples. The primers used for this analysis were shown in Table S19. Data analysis All data were statistically analyzed using the IBM SPSS Statistics 25 software with pairwise comparisons of LSD to identify differences. The data conforming to normal distribution and homogeneity of variance use one-way ANOVA; otherwise Kruskal-Wallis one-way ANOVA was used. Significant differences ( P < 0.05) were indicated by different letters. The GraphPad Prism 8 software was used to draw figures. Abbreviations FHB Fusarium head blight DON Deoxynivalenol GST Glutathione S-transferase GSH Glutathione APM Amiprophosmethyl GISH Genomic in situ hybridization FISH Fluorescence in situ hybridization ND-FISH Non-denaruring fluorescence in situ hybridization hpi Hours post-inoculation DEG Differentially expressed genes PIP5K Phosphatidylinositol 4-phosphate 5-kinase BIP Immunoglobulin-binding protein Hsp Heat shock protein APX Ascorbate peroxidase GR Glutathione reductase HR Hypersensitive response FLS Flavonol synthase CHI Chalcone isomerase CHS Chalcone synthase HCT Hydroxycinnamoyl-coa shikimate MAPK Mitogen-activated protein kinase RBOH Respiratory burst oxidase WGCNA Weighted gene co-expression network analysis PTI Pathogen-associated molecular pattern-trigged immunity PRRs Pattern recognition receptors PLKs Receptor-like kinases PLPs Receptor-like proteins ROS Reactive oxygen species MIK2 MDIS-interacting receptor-like kinase 2 CWIN Cell wall invertase PLC Phospholipase C SA Salicylic acid JA Jasmonate RER Rough endoplasmic reticulum CRT Calreticulin PDI Protein disulfide isomerase PCR Polymerase chain reaction Declarations Acknowledgments We thank the participants for partaking in this study. The authors would like to thank the reviewers whose constructive comments are very helpful for strengthening the presentation of this paper. Authors’ contributions Y.D., data curation, data analysis, visualization, methodology, writing-original draft. W.F., data analysis, material identification. S.C., data curation, investigation of agronomic traits, material planting. J.S., data analysis, material identification, evaluation of fhb resistance. H.L., material planting, material identification. J.L., investigation of agronomic traits, evaluation of FHB resistance. H.M. (Haigang Ma), evaluation of FHB resistance. Y.W., formal analysis, validation. Y.G., visualization. J.Z., and B.W., material radiation. J.C, conceptualization, supervision, data curation. H.M. (Hongxiang Ma), conceptualization, supervision, data curation, funding acquisition, project administration, resources, writing-review & editing. Funding This research was funded by the Project of Zhongshan Biological Breeding (ZSBBL-KY2023-02-3, BM2022008-02), Seed Industry Revitalization Project of Jiangsu Province (JBGS2021047), Jiangsu Key Project for the Research and Development (BE2022346), and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD). Availability of data and materials The genome sequences of Chinese Spring were downloaded from the website: http://www.wheatgenome.org/News2/IWGSC-RefSeq-v2.0-now-available-at-URGI. The genome sequences of Th. elongatum were downloaded from the website: https://ngdc.cncb.ac.cn/gwh/Assembly/965/show. The transcriptome sequencing data generated in this study have been deposited in the NCBI Sequence Read Archive (SRA) database under accession number PRJNA1011388. 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Lei J, Zhou J, Sun H, Wan W, Xiao J, Yuan C, Karafiátová M, Doleel J, Wang H, Wang X. Development of oligonucleotide probes for FISH karyotyping in Haynaldia villosa , a wild relative of common wheat. The Crop J. 2020;8:676-681. Tang Z, Yang Z, Fu S. Oligonucleotides replacing the roles of repetitive sequences pAs1, pSc119.2, pTa-535, pTa 71, CCS1, and PAWRC.1 for FISH analysis. J Appl Genet. 2014;55:313-318. Dai Y, Tabassum MA, Chen L, Pan Z, Song L. Physiological and transcriptomic response of soybean seedling roots to variable nitrate levels. Agronomy J. 2021;113:3639-3652. Additional Declarations No competing interests reported. Supplementary Files Additionalfile1.doc Additionalfile2.xls Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4079736","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":283239444,"identity":"c023329b-291f-4a5a-9bfe-2cb55201e7a9","order_by":0,"name":"Yi Dai","email":"","orcid":"","institution":"Yangzhou University","correspondingAuthor":false,"prefix":"","firstName":"Yi","middleName":"","lastName":"Dai","suffix":""},{"id":283239445,"identity":"16df4ac9-fb3f-46f7-add5-68f8a66adf94","order_by":1,"name":"Wenlin Fei","email":"","orcid":"","institution":"Yangzhou 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Scale bar = 10 cm. (B) GISH analysis of the translocation lines YNM158, diploid \u003cem\u003eTh. elongatum\u003c/em\u003egenomic DNA was used as a probe (green). Arrows show the translocated chromosome pair. Scale bar = 100 μm. (C) ND-FISH analysis of the translocation lines YNM158, Oligo-pAs1 (red signal) modified with 5′TAMRA and Oligo-pSc119.2 (green signal) modified with 5′FAM were used as probes. Chromosomes were counterstained with DAPI (blue). Arrows show the translocated chromosome pairs. Scale bar = 100 μm. (D) Symptoms of YNM158 and the control varieties at 21 dpi with \u003cem\u003eF. graminearum\u003c/em\u003e isolate F0609. Scale bar = 2 cm. Statistical analysis of eight agronomic traits for YNM158 and YM158. Statistical significance of differences was evaluated by t-test (*, \u003cem\u003eP \u003c/em\u003e\u0026lt; 0.05, ***, \u003cem\u003eP\u003c/em\u003e \u0026lt; 0.01)\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-4079736/v1/6b4e33528c60e2e9fc5d55cf.png"},{"id":53554830,"identity":"afa76c63-3365-4dbf-b799-bb20228f8f34","added_by":"auto","created_at":"2024-03-27 12:11:52","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":310401,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAnalysis of the differentially expressed genes (DEGs) identified from YNM158.\u003c/strong\u003e (A) Statistical analysis of the DEGs number on 7EL chromosome at different time after \u003cem\u003eF. graminearum\u003c/em\u003e infection. (B) Gene ontology function enrichment analysis of DEGs on 7EL chromosome after \u003cem\u003eF. graminearum\u003c/em\u003e infection. (C) KEGG pathway enrichment analysis of DEGs on 7EL chromosome after \u003cem\u003eF. graminearum\u003c/em\u003e infection. (D) The heatmap of the DEGs enriched in starch and sucrose metabolism pathway.\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-4079736/v1/2f944c85ccf8f064a11e13ad.png"},{"id":53554828,"identity":"bc8f9ab6-3410-4e2e-9287-730949015795","added_by":"auto","created_at":"2024-03-27 12:11:52","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":446236,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGene expression patterns analysis on 7EL chromosome.\u003c/strong\u003e (A) The trend analysis of DEGs at different times after \u003cem\u003eF. graminearum\u003c/em\u003e infection. (B) Gene dendrogram by clustering the dissimilarity based on topological overlap. (C) Correlation heatmap between modules and infection time with \u003cem\u003eF. graminearum\u003c/em\u003e. The 4 modules are provided in the left panel. The module-trait correlation, from -1 (light blue) to 1 (pink), is indicated with the color scale on the right. Each column presents the infection time, and their association with each module is represented by a correlation coefficient (showing top left corner) and a \u003cem\u003eP\u003c/em\u003e-value (showing lower right corner). (D) Venn diagrams showing the overlapping of DEGs between WGCNA and trend analysis. (E-G) Relative expression of \u003cem\u003eTel7E01G1020600\u003c/em\u003e, \u003cem\u003eTel7E01G946300\u003c/em\u003e and \u003cem\u003eTel7E01G980900 \u003c/em\u003eby qRT-PCR. (H) The correlation analysis between the relative expression obtained by qRT-PCR and RNA-seq by Pearson correlation analysis. *, \u003cem\u003eP \u003c/em\u003e\u0026lt; 0.05, ***, \u003cem\u003eP\u003c/em\u003e \u0026lt; 0.01 by Student’s t-test.\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-4079736/v1/049d6d01610e830fb1ca149c.png"},{"id":53555496,"identity":"0322e57e-2cc4-4c41-877d-0be68828f6f8","added_by":"auto","created_at":"2024-03-27 12:19:52","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":108133,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eScreened the DEGs between YM158 and YNM158\u003c/strong\u003e. (A) DEGs Venn diagram of initial colonization stage. (B) DEGs Venn diagram at infection stage. (C) Different types of specific DEGs statistics.\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-4079736/v1/f22039a81c029189a225f0ab.png"},{"id":53555738,"identity":"51f317d9-56f3-4f9e-aa86-a889c452c506","added_by":"auto","created_at":"2024-03-27 12:27:53","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":793048,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eKEGG pathway enrichment analysis of wheat DEGs.\u003c/strong\u003e (A) The specific enrichment pathways at initial colonization stage. (B) The heatmap of the representative DEGs enriched at initial colonization stage. (C) The specific enrichment pathways at infection stage. (D) The heatmap of the representative DEGs enriched at infection stage. (E) The same enrichment pathways at initial colonization stage. (F) The same enrichment pathways at infection stage. (G) The heatmap of the representative DEGs enriched at both stages.\u003c/p\u003e","description":"","filename":"Figure5.png","url":"https://assets-eu.researchsquare.com/files/rs-4079736/v1/ba792c7791bc28610b677e55.png"},{"id":53555497,"identity":"618ebd4a-4204-4697-8e52-51386bde5bf1","added_by":"auto","created_at":"2024-03-27 12:19:52","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":402218,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eWGCNA of wheat DEGs identified in the YM158 and YNM158 after \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eF. graminearum\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e infection.\u003c/strong\u003e (A) The x-axis represents the soft threshold β. (B) The y-axis represents the mean of all gene’s adjacency functions in the corresponding gene module.(C) Fourteen modules of co-expressed genes are shown in a hierarchical cluster tree. A major tree branch represents a module. Modules in designated colors are presented in the lower panel. (D) Module-trait relationships. The 14 modules are provided in the left panel. The module-trait correlation, from -1 (green) to 1 (red), is indicated with the color scale on the right. The association with each module is represented by a correlation coefficient and a \u003cem\u003eP\u003c/em\u003e-value (showing in parentheses).\u003c/p\u003e","description":"","filename":"Figure6.png","url":"https://assets-eu.researchsquare.com/files/rs-4079736/v1/1a5771c023faf43e32d29369.png"},{"id":53555499,"identity":"5aa978f1-58b1-4ace-abb5-ce477d521653","added_by":"auto","created_at":"2024-03-27 12:19:52","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":441458,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe core genes extraction and expression verification by RT-PCR. \u003c/strong\u003e(A) Venn diagram between the hub genes associated with YNM158 and DEGs in the specific pathways at initial colonization stage. (B) Venn diagram between the hub genes associated with YNM158 and DEGs in the specific pathways at infection stage. (C) DEGs Venn diagram between initial colonization stage and infection stage in the same pathways. (E-J) The relative expression of the core genes in YM158 and YNM158. *, \u003cem\u003eP \u003c/em\u003e\u0026lt; 0.05, ***, \u003cem\u003eP\u003c/em\u003e \u0026lt; 0.01 by Student’s t-test.\u003c/p\u003e","description":"","filename":"Figure7.png","url":"https://assets-eu.researchsquare.com/files/rs-4079736/v1/e97af48f7739e4c967a6e317.png"},{"id":55770172,"identity":"6e835197-27a1-4597-84dd-fef5fa58d355","added_by":"auto","created_at":"2024-05-02 20:56:19","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4059077,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4079736/v1/cfb0ae65-41a7-4519-bd4c-2a241b197881.pdf"},{"id":53554833,"identity":"27792e76-7caf-4453-986d-2bf80ffe4c1d","added_by":"auto","created_at":"2024-03-27 12:11:53","extension":"doc","order_by":12,"title":"","display":"","copyAsset":false,"role":"supplement","size":202685,"visible":true,"origin":"","legend":"","description":"","filename":"Additionalfile1.doc","url":"https://assets-eu.researchsquare.com/files/rs-4079736/v1/191be1e75e074f2c950c6f3f.doc"},{"id":53554837,"identity":"e5e72a4d-c6d1-4d62-8592-75ca2282ce24","added_by":"auto","created_at":"2024-03-27 12:11:58","extension":"xls","order_by":13,"title":"","display":"","copyAsset":false,"role":"supplement","size":109272576,"visible":true,"origin":"","legend":"","description":"","filename":"Additionalfile2.xls","url":"https://assets-eu.researchsquare.com/files/rs-4079736/v1/295cb4c16d8b34a5cbccc050.xls"}],"financialInterests":"No competing interests reported.","formattedTitle":"Transcriptome analysis revealed the potential mechanism of a wheat-Th. elongatum translocation line YNM158 against Fusarium head blight","fulltext":[{"header":"Introduction","content":"\u003cp\u003e \u003cem\u003eFusarium\u003c/em\u003e head blight (FHB) is a primary disease caused by pathogens such as \u003cem\u003eF. asiaticum\u003c/em\u003e and \u003cem\u003eF. graminearium\u003c/em\u003e, seriously affecting yield and quality in wheat worldwide. Wheat infected with FHB, in addition to causing a significant reduction in yield, also accumulates the toxin deoxynivalenol (DON) in the seeds, which poses a significant risk to food safety. With global warming and changes in farming systems and practices, there is a trend of expanding the area of occurrence of the wheat blast, and the cultivation of blast-resistant varieties is the fundamental way to reduce its damage. However, the sources of resistance to wheat FHB were relatively narrow. There were only seven QTLs (\u003cem\u003eFhb1\u003c/em\u003e-\u003cem\u003eFhb8\u003c/em\u003e) related to FHB resistance, such as \u003cem\u003eFhb1\u003c/em\u003e from chromosome 3BS and \u003cem\u003eFhb2\u003c/em\u003e from chromosome 6BS of Sumai 3 [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e], \u003cem\u003eFhb4\u003c/em\u003e from chromosome 4B, \u003cem\u003eFhb5\u003c/em\u003e from chromosome 5A \u003cem\u003eFhb8\u003c/em\u003e from chromosome 7D of Wangshuibai [\u003cspan additionalcitationids=\"CR4\" citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], of which \u003cem\u003eFhb1\u003c/em\u003e was recognized as the most stable master effect QTL for FHB resistance expansion and widely used in wheat breeding for the FHB resistance. Furthermore, the QTLs were also found in wheat relatives, such as \u003cem\u003eFhb3\u003c/em\u003e from \u003cem\u003eLeymus racemosus\u003c/em\u003e [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e], \u003cem\u003eFhb6\u003c/em\u003e from \u003cem\u003eElymus tsukushiensi\u003c/em\u003e [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e] and \u003cem\u003eFhb7\u003c/em\u003e from \u003cem\u003eThinopyrum ponticum\u003c/em\u003e [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAs an essential close relative of wheat, tall wheatgrass has many important favorable traits, which represent a valuable source of alien gene resources for wheat. Studies have shown that there are three species of tall wheatgrass in nature, namely \u003cem\u003eTh. elongatum\u003c/em\u003e (diploid, 2n\u0026thinsp;=\u0026thinsp;2x\u0026thinsp;=\u0026thinsp;14, E\u003csup\u003ee\u003c/sup\u003eE\u003csup\u003ee\u003c/sup\u003e), \u003cem\u003eTh. scirpeum\u003c/em\u003e (tetraploid, 2n\u0026thinsp;=\u0026thinsp;4x\u0026thinsp;=\u0026thinsp;28, E\u003csup\u003ee\u003c/sup\u003eE\u003csup\u003ee\u003c/sup\u003eE\u003csup\u003eb\u003c/sup\u003eE\u003csup\u003eb\u003c/sup\u003e) and \u003cem\u003eTh. ponticum\u003c/em\u003e (decaploid, 2n\u0026thinsp;=\u0026thinsp;10x\u0026thinsp;=\u0026thinsp;70, E\u003csup\u003ee\u003c/sup\u003eE\u003csup\u003ee\u003c/sup\u003eE\u003csup\u003eb\u003c/sup\u003eE\u003csup\u003eb\u003c/sup\u003eE\u003csup\u003ex\u003c/sup\u003eE\u003csup\u003ex\u003c/sup\u003eStStStSt). There is no scientific conclusion on the evolution process of polyploidy of the family in \u003cem\u003eThinopyrum\u003c/em\u003e, but it is believed that many interspecific hybridizations and natural doubling of chromosomes occurred during the evolution process of tall wheatgrass which was similar to common wheat [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Because tall wheatgrass has strong resistance to wheat FHB, so far, breeders have constructed wheat-tall wheatgrass chromosome addition line, substitution line and other genetic materials by distant hybridization using \u003cem\u003eTh. elongatum\u003c/em\u003e and \u003cem\u003eTh. ponticum\u003c/em\u003e, and obtained the results related to FHB resistance. For example, Jauhar et al. created 1E addition lines, 1E (1A) and 1E (1B) diploid substitution lines by crossing durum wheat with diploid \u003cem\u003eTh. elongatum\u003c/em\u003e, and found that the 1E chromosome of diploid \u003cem\u003eTh. elongatum\u003c/em\u003e may carry FHB resistance genes by FHB resistance evaluation [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Liu et al. (2017) obtained a disomic alien addition line with a pair of 7E \u003cem\u003eTh. scirpeum\u003c/em\u003e chromosomes by hybridization of durum cultivar \u0026ldquo;Langdon\u0026rdquo; with the amphiploid 8801 (AABBEE) and found this addition lines showed high resistance to FHB [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Shen et al. identified the FHB resistance in the substitution lines 7E(7A), 7E(7B) and 7D(7E) from \u003cem\u003eTh. elongatum\u003c/em\u003e, and 7el\u003csub\u003e2\u003c/sub\u003e from \u003cem\u003eTh. ponticum\u003c/em\u003e in the Thatcher genetic background [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. This FHB resistant locus, assigned as \u003cem\u003eFhbLoP\u003c/em\u003e, was mapped to the distal region of the long arm of chromosome 7E in \u003cem\u003eTh. ponticum\u003c/em\u003e within a 3.71 cM interval flanked by \u003cem\u003eXcfa2240\u003c/em\u003e and \u003cem\u003eXswes19\u003c/em\u003e, which accounts for 30.46% of the phenotypic variance [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. This locus was then designated as \u003cem\u003eFhb7\u003c/em\u003e and was fine mapped in 1.7 cM interval. Translocation lines with shortened \u003cem\u003eTh. ponticum\u003c/em\u003e chromatin carrying \u003cem\u003eFhb7\u003c/em\u003e was developed [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. After that, Wang et al. [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e] successfully cloned the \u003cem\u003eFhb7\u003c/em\u003e gene from the \u003cem\u003eTh. ponticum\u003c/em\u003e 7el\u003csub\u003e2\u003c/sub\u003e chromosome by assembling the genome of \u003cem\u003eTh. elongatum\u003c/em\u003e. And they proved that this gene encoded a glutathione S-transferase (GST), which can open the epoxy group of DON toxin and catalyze its formation of glutathione adduct (DON-GSH), resulting in detoxification and anti-FHB effect. However, Guo et al. discovered that some wheat-\u003cem\u003eThinopyrum\u003c/em\u003e derivatives carrying the \u003cem\u003eFhb7\u003c/em\u003e homologs had a different reaction to Fusarium head blight [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Similar results were also found in transgenic plants by overexpression of GST-encoding \u003cem\u003eFhb7\u003c/em\u003e [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMoreover, because \u003cem\u003eTh. elongatum, Th. scirpeum\u003c/em\u003e and \u003cem\u003eTh. ponticum\u003c/em\u003e shares the E genome of \u003cem\u003eTh. elongatum\u003c/em\u003e, so previous studies have shown that the FHB resistance genes are also located on the homologous group seven in diploid \u003cem\u003eTh. elongatum\u003c/em\u003e [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. For example, Zhang et al. incorporated a novel \u003cem\u003eFhb7\u003c/em\u003e allele, \u003cem\u003eFhb7\u003c/em\u003e\u003csup\u003e\u003cem\u003eThe2\u003c/em\u003e\u003c/sup\u003e, into the wheat B genome through a small 7B-7E translocation (7BS\u0026middot;7BL-7EL) involving the terminal regions of the long arms, making this novel FHB resistance allele usable for breeding in both common and durum wheat [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIt is well known that the introduction of chromosomes from wild species into wheat usually has linkage drag of undesirable genes and limits their application. In wheat genetic improvement, breeding translocation lines carrying alien beneficial genes, especially small fragment translocation lines, can reduce the linkage caused by alien chromosomes and has high genetic stability under common wheat genetic background, which is an ideal way to introduce alien beneficial genes. Therefore, in this study, we aim to: obtain the translocation lines with different size of 7EL chromosome fragment; develop the stable inheritance translocation line with FHB resistance; evaluate the application value of translocation lines in wheat breeding for FHB resistance; explore potential disease resistance genes and analyze the disease resistance pathways in translocation lines through transcriptome analysis. The results can not only provide new germplasm for wheat FHB resistance breeding, but also provide a theoretical basis for studying the resistance mechanism of wheat FHB.\u003c/p\u003e"},{"header":"Result","content":"\u003cp\u003e\u003cstrong\u003eEstablishment of a wheat-\u003c/strong\u003e \u003cstrong\u003eTh. elongatum\u003c/strong\u003e \u003cstrong\u003e7EL chromosome translocation line YNM158 with FHB resistance\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWhen the alien chromosomes are introduced into wheat, there will be linkage drag, which will have a negative effect on the agronomic characters of wheat. So, creating the small segment translocation lines carrying the alien chromosomes can reduce the negative effects of linkage drag. In this study, the spikes of the translocation line T7BS\u0026middot;7EL, carrying the long arm of diploid \u003cem\u003eTh. elongatum\u003c/em\u003e 7E chromosome, were treated by \u003csup\u003e60\u003c/sup\u003eCo-\u0026gamma; radiation during the flowering period of wheat. After treatment, the fresh pollen was collected and awarded to YM158 to obtain the M\u003csub\u003e1\u003c/sub\u003e seeds. The chromosomes of M\u003csub\u003e1\u003c/sub\u003e generation plants were identified by GISH, and the plants containing 7EL chromosome structure variation was selected for backcross with YM158. Finally, one line with stable agronomic characters was obtained in F\u003csub\u003e6\u003c/sub\u003e generation, which was named YNM158 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eA). The root-tip cells of YNM158 at mitotic metaphase were analyzed by GISH and FISH. Firstly, the presence of the translocation chromosome pair was confirmed in YNM158 by GISH (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eB). Then, the chromosomal structural variation was observed on the end of 4BS chromosome according to the standard karyotype of CS (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eC). Finally, the translocation chromosome can be represented as T7EL\u0026shy;4BS\u0026middot;4BL. In addition, the FHB resistance of YNM158 for two consecutive years showed that YNM158 has a high resistance to FHB in both field and greenhouse, and the level of FHB resistance was similar to that of SU3 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eD and Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Moreover, we observed almost no difference for the tested agronomic traits, including spike length, number of grains per spike, number of spikelets and grain width, for 2 consecutive years (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eE). However, plant height, thousand kernel, weight grain length and flag leaf area of YNM158 were significantly different compared with YM158 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eE).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eMean percentage of diseased spikelets (PDS) in different environments\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eLines\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e2020\u0026ndash;2021\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e2021\u0026ndash;2022\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eField\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eGreenhouse\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eField\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eGreenhouse\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eYNM158\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.81% \u0026plusmn; 0.19d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.18% \u0026plusmn; 0.37c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.21% \u0026plusmn; 0.23d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.27% \u0026plusmn; 0.20d\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSU3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.49% \u0026plusmn; 1.79d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7.38% \u0026plusmn; 2.89c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.36% \u0026plusmn; 0.22d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.06% \u0026plusmn; 0.21d\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAN8455\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e83.42% \u0026plusmn; 10.36a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.83% \u0026plusmn; 14.69a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e87.88% \u0026plusmn; 5.51a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e60.19% \u0026plusmn; 6.24a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eYM23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e28.79% \u0026plusmn; 6.41c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e53.69% \u0026plusmn; 5.15b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e15.54% \u0026plusmn; 3.54c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e21.87% \u0026plusmn; 7.05c\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eYM158\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e47.01% \u0026plusmn; 11.92b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e31.80% \u0026plusmn; 12.91b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e45.09% \u0026plusmn; 8.21b\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctfoot\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"5\"\u003eNote: The data were statistically analyzed by Kruskal-Wallis one-way ANOVA. Pairwise comparisons were completed using LSD. Different letters show significance at \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tfoot\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n\u003ch2\u003eRNA-seq data quality, assembly, and annotation of YNM158 and YM158\u003c/h2\u003e\n\u003cp\u003eTo analyze the genes associated with FHB resistance on chromosome 7EL chromosome segment in YNM158, RNA-sequencing (RNA-seq)-based transcriptome profiling was performed on the spikes which were inoculated F0609. In this experiment, total RNA was extracted from the spikes at 0-hours post-inoculation (hpi, no inoculation), 0.5 hpi, 2 hpi, 8 hpi, 24 hpi, 48 hpi, 72 hpi and 96 hpi, and 24 cDNA libraries were constructed (three repetitions per time). After filtering out the rRNAs and low-quality reads, a total of 193.57 GB high-quality clean data were obtained from 24 libraries (BioProject ID: PRJNA1011388), with an average of 80.65 GB clean data per library. The Q20 and Q30 were \u0026gt;\u0026thinsp;97% and \u0026gt;\u0026thinsp;93%, respectively. In addition, the GC contents were 48.86\u0026ndash;52.53% among all samples. After assembly, the clean reads were mapped to the wheat and \u003cem\u003eTh. elongatum\u003c/em\u003e reference genome (Chinese Spring v2.2 and \u003cem\u003eTh. elongatum\u003c/em\u003e v1.0). On average, 91.17% of the reads were successfully aligned to the reference genome (Table \u003cspan class=\"InternalRef\"\u003eS1\u003c/span\u003e). Therefore, these analyses indicated that the quality of our RNA-seq data was high and the sequencing depth was sufficient for further analysis.\u003c/p\u003e\n\u003cp\u003eTo investigate the impact of chromosome translocations on gene expression, RNA-seq-based transcriptome profiling was also performed on wheat variety YM158 which was one of the parents of YNM158. In this study, we constructed cDNA libraries (24 cDNA libraries) of YM158 after \u003cem\u003eF. graminearum\u003c/em\u003e infection at different time. After filtering out the rRNAs and low-quality reads, a total of 188.56 GB high-quality clean data were obtained from 24 libraries (BioProject ID: PRJNA1011388), with an average of 78.57 GB clean data per library. The Q20 and Q30 were \u0026gt;\u0026thinsp;97% and \u0026gt;\u0026thinsp;94%, respectively. After assembly, the clean reads were mapped to the wheat reference genome (Chinese Spring v2.2). On average, 89.31% of the reads were successfully aligned to the reference genome (Table \u003cspan class=\"InternalRef\"\u003eS1\u003c/span\u003e). Therefore, these analyses indicated that the quality of our RNA-seq data was high and the sequencing depth was sufficient for further analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIdentification of the DEGs on 7EL chromosome post inoculation with\u003c/strong\u003e \u003cstrong\u003eF. graminearum\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBy using the criteria of FDR\u0026thinsp;\u0026lt;\u0026thinsp;0.05, |log2 (fold change)| \u0026gt; 1, a total of 32102 differentially expressed genes (DEGs) were detected that significantly responses to \u003cem\u003eF. graminearum\u003c/em\u003e infection at different times in YNM158, of which 222 DEGs were located on 7EL chromosome (Table \u003cspan class=\"InternalRef\"\u003eS2\u003c/span\u003e). The 222 DEGs were analyzed, and the results showed that there were 60, 10, 25, 27, 49, 109 and 135 DEGs at 0.5 hpi, 2 hpi, 8 hpi, 24 hpi, 48 hpi, 72 hpi and 96 hpi, respectively (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eA). GO function enrichment analysis indicated that these DEGs were enriched in catalytic activity, carboxylic acid transmembrane transporter activity and organic acid transmembrane transporter activity in terms of molecular function. And a total of 124 DEGs were enriched in catalytic activity (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eB, Table S3). In terms of biological processes, organic anion transport, organonitrogen compound catabolic process and anion transport was the main role of the enriched genes. Among them, organonitrogen compound catabolic process was the term with the most enriched DEGs, with a total of 18 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eB, Table S3). And the top 10 GO terms with the lowest Q value were selected to draw the scatter diagram of enrichment items in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eB. In addition, the KEGG pathway analysis revealed that the starch and sucrose metabolism pathway is the only one with the Q value less than 0.05 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eC, Table S4). Moreover, the heatmap of the DEGs enriched in this pathway showed the expression levels of these genes were down-regulated after infection with \u003cem\u003eF. graminearum\u003c/em\u003e, especially in the first 8 hours after infection (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eD). The top 10 pathways with the lowest Q value were selected to draw the scatter diagram of enrichment items in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eC.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n\u003ch2\u003eGene expression patterns analysis of DGEs on 7EL chromosome at different infection time-points\u003c/h2\u003e\n\u003cp\u003eThe trend analysis of 222 DEGs on 7EL chromosome showed that the DEGs were clustered into 20 profiles, of which 153 DEGs were significantly clustered to the three profiles, including profile 19, profile 0 and profile 6, on the basis of \u003cem\u003eP\u003c/em\u003e-value ˂ 0.05 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eA, Table S5). Among them, 77 genes showed a continuous upward trend with the extension of infection time (profile 19), and 56 genes showed a continuous downward trend (profile 0).\u003c/p\u003e\n\u003cp\u003eIn addition, the 222 DEGs were analyzed of the weighted gene co-expression network analysis (WGCNA) modules associated with infection time. The selection of a soft threshold (Power) is the key step to constructing the network. When the soft threshold was set as 10 with a scale-free network fitting index of R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.80, average connectivity was close to 0 (Fig. \u003cspan class=\"InternalRef\"\u003eS1\u003c/span\u003eA). The hierarchical cluster tree was drawn based on the optimal soft threshold, and the genes clustered in the same branch are divided into the same module. Finally, four modules, including turquoise, blue, brown and grey, were obtained (Fig. \u003cspan class=\"InternalRef\"\u003eS1\u003c/span\u003eB). Among them, the brown module was significantly correlated with infection at 0.5 hpi (R\u0026thinsp;=\u0026thinsp;0.92, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;0.05), the turquoise module was significantly correlated with infection at 8 hpi (R\u0026thinsp;=\u0026thinsp;0.82, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;0.05), and the blue module was significantly correlated with infection at 96 hpi (R\u0026thinsp;=\u0026thinsp;0.69, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;0.05). Based the cut-off criteria |module member-ship| (|MM|)\u0026thinsp;\u0026gt;\u0026thinsp;0.8 and |gene significance| (|GS|)\u0026thinsp;\u0026gt;\u0026thinsp;0.8, 19 genes with high connectivity in the clinically significant module were identified as hub genes (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eB and \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eC, Table S6).\u003c/p\u003e\n\u003cp\u003eThe DEGs and hub genes obtained from trend analysis and WGCNA, respectively, were sued to draw the Venn diagrams, and a total of 12 genes were obtained in both analyses (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eD, Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). The qRT-PCR verification of 12 DEGs showed that the relative expression levels of several genes were induced by \u003cem\u003eF. graminearium\u003c/em\u003e, such as \u003cem\u003eTel7E01G1020600\u003c/em\u003e, \u003cem\u003eTel7E01G943900\u003c/em\u003e and \u003cem\u003eTel7E01G980900\u003c/em\u003e. Among them, \u003cem\u003eTel7E01G1020600\u003c/em\u003e encoded glutathione S-transferase (GST), whose expression was induced by \u003cem\u003eF. graminearium\u003c/em\u003e and significantly upregulated from 72 hpi (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eE). \u003cem\u003eTel7E01G943900\u003c/em\u003e and \u003cem\u003eTel7E01G1980900\u003c/em\u003e had similar expression patterns, both of which were up-regulated at the early stage of infection. \u003cem\u003eTel7E01G943900\u003c/em\u003e encodes a receptor-like kinase, and the expression of this gene was up-regulated by about 5 times at 0.5 hpi, but then began to be down-regulated (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eF). Similarly, \u003cem\u003eTel7E01G1980900\u003c/em\u003e encoded a monosaccharide-sensing protein, the expression of this gene was significantly reached the highest level at 2 hpi, which was about 4 times than that at 0 hpi, but then began to be down-regulated and was almost no expression after 24 h of infection (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eG). These results suggested that there may be multiple resistance genes on chromosome 7EL fragment, which provide resistance to FHB at different stages of \u003cem\u003eF. graminearium\u003c/em\u003e infection.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eThe gene was verified by qRT-PCR.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eGene ID\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eGene description\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G1002700\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eLysine ketoglutarate reductase trans-splicing-like protein (DUF707)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G1013700\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eGalactoside 2-alpha-L-fucosyltransferase\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G1020600\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eGlutathione S-transferase\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G211400\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eProtein kinase\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G899900\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eNF-X1-type zinc finger protein NFXL1\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G905000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eDisease resistance protein (NBS-LRR class) family\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G934300\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCarbonic anhydrase\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G939300\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eReceptor-like kinase\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G941500\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCarboxypeptidase\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G943900\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eReceptor-like kinase\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G946300\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eBlue copper binding protein\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTel7E01G980900\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eMonosaccharide-sensing protein 2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eExtraction of wheat DEGs between YM158 and YNM158 after\u003c/strong\u003e \u003cstrong\u003eF. graminearum\u003c/strong\u003e \u003cstrong\u003einfection\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSince wheat FHB is a compatible disease, Stephens et al., (2008) divided the infection process into initial colonization stage, infection stage and late infection stage [\u003cspan class=\"CitationRef\"\u003e20\u003c/span\u003e]. In this study, we analyzed the effect of the introduction of 7EL chromosome fragments on FHB resistance using 8 hpi as the cut-off point. And we defined the initial colonization stage before 8hpi, the infection stage after 8hpi. First of all, 12761 and 15719 DEGs were obtained in the initial colonization stage and the infection stage, respectively (Table S7 and S8). Then, the DEGs at 0 hpi between YM158 and YNM158 were removed (Table S9). Finally, a total of 4734 DEGs were obtained at the initial stage of colonization (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003eA), and 10489 DEGs were obtained at the stage of infection (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003eB). After alignment with the reference genome, 4060 and 9808 wheat DEGs were screened for subsequent analysis at the initial colonization stage and infection stage, respectively (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003eC, Table S10).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n\u003ch2\u003eKEGG pathway enrichment analysis of wheat DEGs at different stages\u003c/h2\u003e\n\u003cp\u003eTo analyze the effect of 7EL chromosome on the resistance pathway of wheat at different stages after \u003cem\u003eF. graminearum\u003c/em\u003e infection, the wheat DEGs were subjected to KEGG pathway enrichment analysis. We identified 7 pathways that were significantly enriched with a Q value less than 0.05 during the initial colonization stage, among which phosphatidylinositol signaling system was the most enriched pathway with the lowest Q value and the protein processing in endoplasmic reticulum pathway was the most enriched pathway with the DEGs number (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eA, Table S11). Further analysis of the genes involved in these pathways was found that the expression of some genes related to phosphatidylinositol 4-phosphate 5-kinase (PIP5K), immunoglobulin-binding protein (BIP4) and heat shock protein (Hsp) in YNM158 was higher than that in YM158 after \u003cem\u003eF. graminearum\u003c/em\u003e infection (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eB).\u003c/p\u003e\n\u003cp\u003eAt the infection stage, 21 specific pathways were significantly enriched with a Q value less than 0.05. Among them, glutathione metabolism was the pathway with the lowest Q value and the largest number of DEGs (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eC, Table S11). Moreover, genes related to ascorbate peroxidase (APX), glutathione reductase (GR), glutathione-S-transferase (GST) in this pathway were up-regulated in YNM158 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eD). In addition, we also found that the genes related to ABC transporter (ATP-binding cassette, ABC) were up-regulated in YNM158, they may be associated with the transportation of DON (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eD). The plant-pathogen interaction pathway was the second most enriched pathway, with a total of 152 DEGs enriched in this pathway (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eC). Through the heat map of gene expression, we found that the expression levels of some genes related to hypersensitive response (HR), such as \u003cem\u003eTraesCS2D03G0030700\u003c/em\u003e and \u003cem\u003eTraesCS2D03G1070500\u003c/em\u003e, began to be up-regulated at 24 hpi with \u003cem\u003eF. graminearum\u003c/em\u003e in YNM158 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eD).\u003c/p\u003e\n\u003cp\u003eIt is well known that secondary metabolites play an important role in plant resistance to pathogens. In our study, a total of 13 pathways were identified in both initial colonization stage and infection stages with a Q value less than 0.05. Among them, biosynthesis of secondary metabolites was the pathway with the lowest Q value (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eE and \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eF, Table S12). The analysis of gene expression in this pathway found that some genes related to flavonol synthase (FLS), chalcone isomerase (CHI), chalcone synthase (CHS) and hydroxycinnamoyl-CoA shikimate (HCT) showed a significantly up-regulated expression trend with the extension of \u003cem\u003eF. graminearum\u003c/em\u003e infection time in YNM158 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eG). In addition, in the MAPK pathway, we also found the expression of some genes related to respiratory burst oxidase (RBOH) was significantly up-regulated after \u003cem\u003eF. graminearum\u003c/em\u003e infection, and the expression level was the highest at 72 hpi, while the expression of some genes related to mitogen-activated protein kinase (MAPK) was up-regulated at initial colonization stage and then down-regulated at the infection stage in YNM158 after \u003cem\u003eF. graminearum\u003c/em\u003e infection in YNM158 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eG).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n\u003ch2\u003eWGCNA of wheat DEGs\u003c/h2\u003e\n\u003cp\u003eThe 12661 wheat DEGs were also analyzed of the WGCNA modules (Table S13). The selection of a soft threshold (Power) is the key step to constructing the network. When the soft threshold was set as 9 with a scale-free network fitting index of R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.80, average connectivity was close to 0 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003eA and \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003eB). The hierarchical cluster tree was drawn based on the optimal soft threshold, and the genes clustered in the same branch are divided into the same module (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003eC). Finally, 14 modules, which were correlated with varieties were obtained. Among them, the yellow module was significantly positive associated with YNM158 (R\u0026thinsp;=\u0026thinsp;0.96, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;0.05), and a total of 847 DEGs were obtained in this module (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003eD, Table S14).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n\u003ch2\u003eScreened the core genes at different infection stages\u003c/h2\u003e\n\u003cp\u003eThe Venn diagrams was drawn between the wheat DEGs at different infection stages and the hub genes positive associated with YNM158. At initial colonization stage, a total of 120 DEGs were obtained from the specific enrichment pathways (Table S15), and among them, 13 DEGs may be associated with the FHB resistance of YNM158 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003eA, Table S16). At infection stage, 830 DEGs were obtained from the specific pathway (Table S17), of which 10 DEGs may be related to the FHB resistance of YNM158 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003eB, Table S16). In addition, among the same pathways in both stages mentioned above, we obtained 202 DEGs (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003eC, Table S18), 21 of which may be related the FHB resistance of YNM158 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003eD, Table S16).\u003c/p\u003e\n\u003cp\u003eIn order to verify the correlation between the core genes and the stages of FHB resistance, these selected genes were verified by RT-PCR. Compared with the expression of YM158 at 0 hpi, we found that the expression levels of 6 wheat genes in YNM158 were significantly up-regulated after \u003cem\u003eF. graminearum\u003c/em\u003e infection (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). Among them, \u003cem\u003eTraesCS4D03G0528700\u003c/em\u003e and \u003cem\u003eTraesCS4B03G0573000\u003c/em\u003e belong to the phosphatidylinositol signaling system and protein processing in endoplasmic reticulum pathway, respectively. And had the same expression pattern at initial colonization stage, both of which were significantly up-regulated in YNM158. \u003cem\u003eTraesCS4D03G0528700\u003c/em\u003e had the highest expression level at 2 hpi (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003eE) and the expression of \u003cem\u003eTraesCS4B03G0573000\u003c/em\u003e reached the highest level at 8 hpi (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003eF). \u003cem\u003eTraesCS2D03G0030700\u003c/em\u003e and \u003cem\u003eTraesCS7D03G0466200\u003c/em\u003e encoded NBS-LRR disease resistance protein and 3-ketoacyl-CoA synthase, respectively, both belong to the plant-pathogen interaction pathway. However, their expression patterns were slightly different. The expression of \u003cem\u003eTraesCS2D03G0030700\u003c/em\u003e was significantly up-regulated after \u003cem\u003eF. graminearum\u003c/em\u003e infection and the expression of this gene in YNM158 was always higher than that of YM158 (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003eG). Although \u003cem\u003eTraesCS7D03G0466200\u003c/em\u003e was also induced after \u003cem\u003eF. graminearum\u003c/em\u003e infection, the expression of \u003cem\u003eTraesCS7D03G0466200\u003c/em\u003e in YNM158 was significantly higher than that in YM158 until 48 hpi (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003eH). In addition, we found that the expression of one gene in biosynthesis of secondary metabolites and MAPK signaling pathway-plant was induced by \u003cem\u003eF. graminearum\u003c/em\u003e. The results of RT-PCR indicated that these two genes may mediate the resistance of YNM158 to FHB during the initial colonization stage and infection stage. \u003cem\u003eTraesCS7A03G1308100\u003c/em\u003e encoded hydroxycinnamoyl-CoA shikimate, the expression of this gene in YNM158 was significantly higher than that in YM158 from 8 hours after \u003cem\u003eF. graminearum\u003c/em\u003e infection (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003eI). On the contrary, the expression of \u003cem\u003eTraesCS1A03G0718100\u003c/em\u003e was significantly up-regulated at 0.5 hpi in YNM158. However, there was little difference in the expression of this gene between YM158 and YNM158 after 48 hpi (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003eJ). The above results suggest that the introduction of 7EL chromosome fragments may affect the disease resistance pathway of wheat, thereby improving the FHB resistance of wheat.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab3\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eThe core genes verified by RT-PCR at different infection stages.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eGene ID\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePathway\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eGene description\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eInitial colonization stage\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTraesCS4D03G0528700\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePhosphatidylinositol signaling system\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePhosphatidylinositol-4-phosphate 5-kinase family protein\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTraesCS4B03G0573000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eProtein processing in endoplasmic reticulum\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70 kDa heat shock protein\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eInfection stage\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTraesCS2D03G0030700\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePlant-pathogen interaction\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eNBS-LRR disease resistance protein\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTraesCS7D03G0466200\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePlant-pathogen interaction\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3-ketoacyl-CoA synthase\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eBoth stage\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTraesCS7A03G1308100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eBiosynthesis of secondary metabolites\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eHydroxycinnamoyl-CoA\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTraesCS1A03G0718100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eMAPK signaling pathway - plant\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eRespiratory burst oxidase-like protein\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eYNM158 can be effectively applied in FHB improvement in wheat breeding\u003c/h2\u003e \u003cp\u003eBreeding and applying FHB resistant varieties in wheat production is an effective way to control the destructive disease. However, the long-term intraspecific cross breeding of wheat reduced the range of genetic variation among varieties and had poor resistance to FHB, while the related wild species and genera of wheat carry many FHB resistance genes. For example, the FHB resistance genes \u003cem\u003eFhb3\u003c/em\u003e, \u003cem\u003eFhb6\u003c/em\u003e and \u003cem\u003eFhb7\u003c/em\u003e on chromosome 7Lr#1S of \u003cem\u003eLeymus racemosus\u003c/em\u003e, chromosome 1E(ts)#1S of \u003cem\u003eElymus tsukushiensis\u003c/em\u003e and chromosome 7el\u003csub\u003e2\u003c/sub\u003e of \u003cem\u003eTh. ponticum\u003c/em\u003e, respectively were reported to have major resistance to FHB [\u003cspan additionalcitationids=\"CR7 CR8\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. In addition, the 1Y\u003csup\u003ec\u003c/sup\u003e and 3S\u003csup\u003ec\u003c/sup\u003e chromosomes of \u003cem\u003eRoegneria ciliaris\u003c/em\u003e [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e], the 1E, 7E chromosome of diploid \u003cem\u003eTh. elongatum\u003c/em\u003e, the chromosome 3St of \u003cem\u003eElymus repens\u003c/em\u003e [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], and the 7M\u003csup\u003eg\u003c/sup\u003e chromosome of \u003cem\u003eAegilops geniculata\u003c/em\u003e [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] also possessed the FHB resistance genes. Although the introduction of alien chromosomes can improve the resistance to FHB, it also brings some genetic encumbrance, which makes the agronomic characters of most foreign germplasm poor and difficultly to be directly used in wheat breeding for FHB resistance. Therefore, to make full use of wheat related species in wheat breeding for FHB resistance, it is necessary to create small fragment translocation lines to develop new varieties with increased FHB resistance and no yield penalty. At present, \u003cem\u003eFhb7\u003c/em\u003e gene from \u003cem\u003eTh. ponticum\u003c/em\u003e was poured into cultivated wheat through small segment translocation lines and used in wheat breeding for FHB resistance [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. The diploid form of tall wheatgrass, \u003cem\u003eTh. elongatum\u003c/em\u003e, has a high level of resistance to FHB and was used to increase FHB resistance in wheat cultivar Chinese Spring by translocation development [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. In this study, we have successfully established the translocation lines with the small fragments of chromosome 7EL from diploid \u003cem\u003eTh. elongatum\u003c/em\u003e by physical radiation, one of which has excellent agronomic traits and high resistance to wheat FHB and named YNM158. Different from previous reports, the translocation in YNM158 occurred on chromosome 4BS, a non-compensative translocation, and the reason for this phenomenon was that chromosome translocation was induced by ionizing radiation, the breakage and reconnection of wheat and alien chromosomes were random, so most of them were uncompensated translocations. Interestingly, the survey results of agronomic traits for two consecutive years showed that YNM158 did not have any bad performances due to a non-compensatory translocation line, such as genetic instability of exogenous chromosomes, high plant height and poor fertility. This may be because a small alien chromosome fragment was attached to the distal end of 4BS chromosome in recurrent parent in YNM158, whereas no chromosome fragments were lost in wheat. And previous studies have also shown that non-compensatory translocation lines also have important utility in wheat breeding. For example, the small fragment translocation line 5VS-6AS\u0026middot;6AL can be used to improve the quality of wheat soft grains [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e], the 3A-7J\u003csup\u003es\u003c/sup\u003e translocation line can be used to improve wheat stem rust resistance [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e], two homozygous translocation lines T1AS\u0026middot;1AL-6VS and T4BS\u0026middot;4BL-6VS-4BL carrying \u003cem\u003ePm21\u003c/em\u003e can be used to enhance powdery mildew resistance in wheat [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Therefore, we propose the translocation line YNM158, contained small fragment of 7EL chromosomes, has a good application prospect in the breeding for resistance to FHB in wheat.\u003c/p\u003e \u003cp\u003e \u003cb\u003eTranscriptome analysis validated that glutathione metabolic is one of the important contributors to FHB resistance in pathogen infection stage\u003c/b\u003e \u003c/p\u003e \u003cp\u003eTranscriptome analysis is a valuable tool in investigating the molecular mechanisms behind cereal resistance to fungal infections as the costs of this technique decrease and its applications become more widespread. In this study, we found that compared with YM158, the glutathione metabolic was the most significantly enriched pathway in YNM158 after \u003cem\u003eF. graminearum\u003c/em\u003e infection during the infection stage (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eD). It is well known that plants respond to fungal infections by activating defense genes including producing reactive oxygen species (ROS) which can enhance the strength of plant cell wall to resist the invasion and colonization of pathogens [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. However, when ROS accumulates in large amounts in plants, it will cause oxidative stress, damage plant cells, and lead to cell dysfunction and even death. Currently, the enzymes involved in the antioxidant defense system can be divided into two groups: (i) Enzymatic antioxidants such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase GPX, glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), and dehydroascorbate reductase (DHAR); (ii) Non-enzymatic antioxidants such as ascorbic acid (AA), reduced glutathione (GSH), α-tocopherol, carotenoids, plastoquinone/ubiquinone and flavonoids [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Early research reported that ascorbate-glutathione (AsA-GSH) cycle is the important way to diameter of removal of ROS in plant. Among them, APX is the key enzymes of this cycle, which can utilize AsA as the electron donor reducing H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e to water, and prevents the accumulation of a toxic level of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e in photosynthetic organisms under stress conditions [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. And the glutathione (GSH) participate in various metabolic processes and were the essential components of antioxidative and detoxification systems in plant cells [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. It can be used as both a reducing agent and a strong nucleophile, participating in the elimination of reactive oxygen species (ROS) through thiol-disulphide redox reactions, and in the detoxification of various heterogenic organisms through conjugation reactions, respectively [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. However, GSH will be oxidized to oxidized glutathione (GSSG) in the reaction of clearing ROS. In order to maintain the balance of GSH content in the plant, GR enzyme will effectively and timely reduce GSSG to GSH. It can be seen that GR enzyme plays a very important role in clearing ROS and maintaining the content of GSH in plants. For example, overexpression of \u003cem\u003eGR\u003c/em\u003e gene from \u003cem\u003eHaynaldia villosa\u003c/em\u003e in wheat can increase resistance to powdery mildew [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. In this study, we found that the expression of some APX-encoding and GR-encoding genes in YNM158 was up-regulated during the infection stage (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eD). Therefore, we suggest that, glutathione may play a key role in ROS-mediated resistance to FHB in wheat.\u003c/p\u003e \u003cp\u003eIn addition, glutathione S-transferase (GST) represents a group of multifunctional enzymes widely present in plants and plays important roles in plant secondary metabolism [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e], growth and development [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e], and biotic and abiotic stress responses [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. Furthermore, one of its most important functions is the ability to inactivate toxic compounds. Because GST can form complexes with glutathione (GSH) by catalyzing hormones, toxins to inactivate or eliminate toxicity of many substances, and expel them in the body under the action of relevant transporters [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. These results suggested that GST played a crucial role in plant disease resistance. For example, \u003cem\u003eNbGSTU1\u003c/em\u003e can increase the resistance to \u003cem\u003eColletotrichum destructivum\u003c/em\u003e in \u003cem\u003eNicotiana benthamiana\u003c/em\u003e [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. The lack of \u003cem\u003eGSTU13\u003c/em\u003e function resulted in enhanced disease susceptibility toward several fungal pathogens in \u003cem\u003eArabidopsis thaliana\u003c/em\u003e [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. Overexpression of \u003cem\u003eLrGST5\u003c/em\u003e in tobacco can improve the resistance of transgenic plants to \u003cem\u003eF. oxysporum\u003c/em\u003e [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. \u003cem\u003eTaGSTU6\u003c/em\u003e interactions can enhance wheat resistance to powdery mildew but not wheat stripe rust [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. We know that wheat infected with FHB can be contaminated with a variety of mycotoxins, especially deoxynivalenol (DON) [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. It has been reported that GSH can form a GSH-DON conjugates under the catalysis of GST to reduce the accumulation of DON and protect plants from toxicity. For instance, \u003cem\u003eFhb7\u003c/em\u003e and \u003cem\u003eFhbRc1\u003c/em\u003e, encoding glutathione S-transferase, enhanced the resistance to FHB in wheat background [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. In this study, the expression of GST- encoding genes, including \u003cem\u003eTraesCS1A03G0109100\u003c/em\u003e, \u003cem\u003eTraesCS3D03G0946300\u003c/em\u003e, \u003cem\u003eTraesCS4D03G0493500\u003c/em\u003e, \u003cem\u003eTraesCS5B03G0050700\u003c/em\u003e, \u003cem\u003eTraesCS5A03G0730500\u003c/em\u003e, \u003cem\u003eTraesCS5B03G0770700\u003c/em\u003e and \u003cem\u003eTel7E01G1020600\u003c/em\u003e was significantly up-regulated after infection with \u003cem\u003eF. graminearum\u003c/em\u003e in YNM158. Among them, the expression of the \u003cem\u003eFhb7\u003c/em\u003e homolog \u003cem\u003eTel7E01G1020600\u003c/em\u003e increased sharply at 72 hpi, which was tens of times higher than that of the non-infection (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eE). It can be seen that GST is one of the important contributors to FHB resistance roles in pathogen infection stage. However, the \u003cem\u003eTel7E01G1020600\u003c/em\u003e in YNM158 was derived from diploid \u003cem\u003eTh. elongatum\u003c/em\u003e, which is not consistent with the origin of Fhb7. So, whether \u003cem\u003eTel7E01G1020600\u003c/em\u003e in YNM158 has the same disease resistance function as \u003cem\u003eFhb7\u003c/em\u003e needs to be further studied.\u003c/p\u003e \u003cp\u003e \u003cb\u003eOther genes from 7EL fragment in YNM158 might also be involved in increasing FHB resistance especially in pathogen initial colonization stage\u003c/b\u003e \u003c/p\u003e \u003cp\u003eDON toxin is a very important fungal pathogen when \u003cem\u003eF. graminearum\u003c/em\u003e infects wheat. It can synthesize a large amount of \u003cem\u003eF. graminearum\u003c/em\u003e along the inflorescence axis and promote the process of disease expansion. However, some studies have reported that when the pathogen initially infected wheat anthers, there was no DON synthesis signal, and only when the disease spread along the inflorescence axis from the inoculation point, DON began to be synthesized in large quantity in the pathogen [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e, \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. It can be seen that DON can help the pathogen spread along the wheat spike axis, but it is not necessary for its initial infection [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. In the process of long-term co-evolution between plants and pathogens, a series of complex defense mechanisms have gradually formed. Generally, pathogen-associated molecular pattern (PAMP)-trigged immunity (PTI) is the first defensive line of plant innate immunity and is mediated by pattern recognition receptors (PRRs). And the PRRs are divided into two types, receptor-like kinases (PLKs) and receptor-like proteins (PLPs). To date, many PLKs have been found to play a key role in wheat disease resistance. For example, Sun et al. (2023) reported that a repeat receptor-like kinase-encoding gene \u003cem\u003eTaBIR1\u003c/em\u003e contributed to wheat resistance against \u003cem\u003ePuccinia striiformis f. sp. tritici\u003c/em\u003e by mediating ROS production and callose deposition [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e], and the cysteine-rich receptor-like kinase TaCRK3 contributed to defense against \u003cem\u003eRhizoctonia cerealis\u003c/em\u003e in wheat through directing antifungal activity and heightening the expression of defense-associated genes in the ethylene signaling pathway [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. And the RLKs have also been found to contribute to grain resistance to \u003cem\u003eFusarium\u003c/em\u003e resistance in cereals. For instance, Thapa et al. (2018) identified two homologous genes on barley chromosome 6H (HvLRRK-6H) and wheat chromosome 6DL (TaLRRK-6D), respectively, which could enhance cereal resistance to FHB disease [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e]. And the Arabidopsis senses \u003cem\u003eFusarium\u003c/em\u003e elicitors in early immune responses to extracts from \u003cem\u003eFusarium\u003c/em\u003e spp. by a novel receptor complex which was encoded by the leucine-rich repeat receptor-like kinase MDIS1-interacting receptor-like kinase 2 (MIK2) at the cell surface [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. Interesting, we also identified several RLKs-encoding genes on the 7EL fragment in this study, and the expression of them was significantly up-regulated at initial colonization stage after \u003cem\u003eF. graminearum\u003c/em\u003e inoculation, such as the expression of \u003cem\u003eTel7E01G943900\u003c/em\u003e which was significantly up-regulated at 0.5 hpi (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eF).\u003c/p\u003e \u003cp\u003eIn addition, in immune responses, plants have developed a number of disease-resistance mechanisms to resist nutrient uptake by pathogens, which involve sugar transport, metabolism, and signal transduction. The previous studies have shown that hexose released by cell wall invertase (CWIN) activity not only acts as a signal molecule to trigger the expression of disease-resistance related genes, but also is an essential metabolite and energy source for the synthesis of antioxidant compounds and defense molecules, such as salicylic acid and callose [\u003cspan additionalcitationids=\"CR53\" citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. For example, \u003cem\u003eAtSTP4\u003c/em\u003e and \u003cem\u003eAtβfruct1\u003c/em\u003e encoding monosaccharide transporter and CWIN, respectively, are both induced in \u003cem\u003eArabidopsis\u003c/em\u003e during parasitic infection by fungus [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e]. Chang et al. (2020) reported that silence the hexes transporter-encoding gene \u003cem\u003ePsHXT1\u003c/em\u003e in wheat stripe rust can significantly inhibit the pathogenicity of pathogenic bacteria [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e]. In this study, the expression of a monosaccharide-sensing protein-encoding gene \u003cem\u003eTel7E01G980900\u003c/em\u003e was significantly up-regulated within 8h after infection with \u003cem\u003eF. graminearum\u003c/em\u003e and reached the highest level at 2 hpi in YNM158 (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eG). And knowledge of the function of monosaccharide-sensing protein is similar to that of hexose transporters [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e, \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e]. Therefore, we speculated that they can also be conducted with CWINs to bring hexose back to host cells, reducing sugar availability to the pathogen, and thus improve host disease resistance. But this needs to be confirmed in the further studies.\u003c/p\u003e \u003cp\u003eIt is well known that \u003cem\u003eF. graminearum\u003c/em\u003e is a kind of facultative trophic bacteria. Therefore, wheat needs to use a series of defense mechanisms to resist pathogen infection at different stages. The introduction of 7EL chromosome fragments not only brought GST-encoding gene which was one of the important contributors on DON detoxification, but also brought other genes which were up-regulated at initial colonization stage. And these genes were also involved in increasing FHB resistance. Therefore, in-depth study of these genes can provide new insights into the molecular mechanisms of wheat resistance to FHB.\u003c/p\u003e \u003cp\u003e \u003cb\u003eIntrogression of 7EL fragment altered the gene expression in wheat after\u003c/b\u003e \u003cb\u003eF. graminearum\u003c/b\u003e \u003cb\u003einoculation\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe introduction of alien chromosome fragments not only brought the resistance genes, but also affects gene expression on normal chromosomes [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e, \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. In this study, we also found that the translocation of chromosomes affected the expression of wheat genes which were enriched in the resistance pathways including phosphatidylinositol signaling system, protein processing in endoplasmic reticulum, plant-pathogen interaction and MAPK signaling pathway at different stages with \u003cem\u003eF. graminearium\u003c/em\u003e infection.\u003c/p\u003e \u003cp\u003eWhen plants are infected with pathogens, phospholipase C (PLC) is rapidly activated by different pathogen-associated molecular patterns (PAMPs) and effector proteins in plant cells [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e]. And then catalyze phosphatidylinositol 4-phosphate (PI4P) and phosphatidylinositol (4,5) bisphosphate [PI(4,5)P2] to produce inositol 2-phosphate (IP2) or inositol 3-phosphate (IP3) and diacylglycerol (DAG). These are conserved compounds of pathogenic microbes that are perceived by immune receptors present in resistant plants [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e, \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e]. The previous studies reported that silencing and knock‑out \u003cem\u003eSlPLC2\u003c/em\u003e in tomato can reduce susceptibility to \u003cem\u003eBotrytis cinereal\u003c/em\u003e [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e, \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e]. And the \u003cem\u003eSlPLC6\u003c/em\u003e plays a key role in both for Ve1 resistance protein mediated resistance to \u003cem\u003eVerticillium dahliae\u003c/em\u003e and Pto/Prf resistance protein mediated resistance to \u003cem\u003ePseudomonas syringae\u003c/em\u003e [\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e]. In addition, it has been reported that salicylic acid (SA), jasmonate (JA)and methyl jasmonate can increase the expression of \u003cem\u003eOsPI-PLC\u003c/em\u003e in rice (\u003cem\u003eOryza sativa\u003c/em\u003e) and improve the resistance of rice to \u003cem\u003eMagnaporthe oryzae\u003c/em\u003e [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e]. In this study, it was found that the expression of some \u003cem\u003ePLC\u003c/em\u003e genes in YNM158 was higher than that in YN158 at the initial colonization stage, such as \u003cem\u003eTraesCS4A03G0225500\u003c/em\u003e and \u003cem\u003eTraesCS4B03G0547100\u003c/em\u003e. Moreover, the results of qRT-PCR showed that the expression of \u003cem\u003eTraesCS4D03G0528700\u003c/em\u003e, which encoded phosphatidylinositol 4-phosphate-5 kinase (PIPK5), in YNM158 was higher than that in YM158 at initial colonization stage (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eB). We know that PIP5K was the catalytic enzyme for the synthesis of PI(4,5)P2 and Shimada et al., (2019) have pointed out that the biosynthesis of PI(4,5)P2 was an important target to improve the defense ability of \u003cem\u003eArabidopsis thaliana\u003c/em\u003e against Colletotrichum, and its activity also determines the defense ability of \u003cem\u003eArabidopsis thaliana\u003c/em\u003e against Colletotrichum [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e]. Therefore, we speculated that PIP5K gene can affect the accumulation of PI(4,5)P2 in YNM1158 to participate in the PLC-mediated response to \u003cem\u003eF. graminearium\u003c/em\u003e infection, and thus affect the colonization of \u003cem\u003eF. graminearium\u003c/em\u003e to improve the resistance to FHB during the initial stages of infection.\u003c/p\u003e \u003cp\u003eThere are also defense-related proteins in plants that are synthesized by the rough endoplasmic reticulum (RER), so when plants attacked by the pathogen, the genes encoding endoplasmic reticulum (ER) chaperones are induced, such as the immunoglobulin-binding protein (BIP), heat shock protein (Hsp), calreticulin (CRT) and protein disulfide isomerase (PDI)-encoding genes. The previous studies have shown that the Hsp was one of the ER chaperones, which play an indispensable role as molecular chaperones in the quality control of PRRs and intracellular resistance (R) proteins against potential invaders [\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e]. For example, Hsp90 was not only involved in the defense of many microbial pathogens by activating the cytosolic R proteins containing nucleotide-binding domain and a leucine-rich repeat, but also participated in chitin responses and anti-fungal immunity in a chaperone complex with its co-chaperone Hop/Sti1 [\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e, \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. In terms of specific diseases, cytoplasmic \u003cem\u003eCapsicum annuum\u003c/em\u003e Hsp70 (CaHsp70) can enhance the resistance to \u003cem\u003eXanthomonas campestris\u003c/em\u003e pv. \u003cem\u003evesicatoria\u003c/em\u003e in pepper [\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e], GmHsp40 can increase soybean resistance to Soybean \u003cem\u003emosaic virus\u003c/em\u003e [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e], Hsp70 can enhance the resistance to powdery mildew in cucumber under heat shock-induction [\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e], MeHsp90.9-MeSGT1-MeRAR1 chaperone complex interacted with MeATGs to trigger autophagy signaling to improve disease resistance to cassava bacterial blight [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e]. In this study, we found the expression of \u003cem\u003eTraesCS4B03G0573000\u003c/em\u003e, which encodes the heat shock protein in YNM158 significantly induced to be upregulated after infection by \u003cem\u003eF. graminearium\u003c/em\u003e at initial colonization stage, which was opposite to the expression pattern in YM158 (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eF). Usually, the expression of the genes encoding ER chaperones even predates the expression of genes encoding pathogenesis-related (PR) proteins [\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e]. Therefore, we inferred that the expression of genes encoding Hsp protein would rapidly induce after infection with \u003cem\u003eF. graminearium\u003c/em\u003e in YNM158, thus activating the defense mechanism earlier, inducing programmed cell death, affecting the colonization of pathogens, and making plants resistant to disease, which provides a new idea for further research on the mechanism of FHB resistance.\u003c/p\u003e \u003cp\u003eReactive oxygen species (ROS) are the important signaling molecules in defense responses during plant-pathogen interactions, which are mainly produced by respiratory burst oxidase homologs (RBOHs) [\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e]. In Arabidopsis, \u003cem\u003eAtRBOHD\u003c/em\u003e and \u003cem\u003eAtRBOHF\u003c/em\u003e are responsible for ROS production against pathogen attacks [\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e]. In \u003cem\u003eNicotiana benthamiana\u003c/em\u003e, \u003cem\u003eNbRBOHA\u003c/em\u003e and \u003cem\u003eNbRBOHB\u003c/em\u003e silencing leads to less ROS production and reduced the resistance against the infection by potato pathogen \u003cem\u003ePhytophthora infestans\u003c/em\u003e [\u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e76\u003c/span\u003e]. Phosphorylation is known to be one of the essential mechanisms of RBOHD activation and is also transcriptionally activated by some kinases, such as MAPK cascades, and the transcriptional regulation of RBOHs may play a key roles in subsequent ROS bursts after turnover of the plasma membrane-localized RBOHs used for the first burst [\u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e77\u003c/span\u003e]. For example, Yamamizo et al. (2007) reported that MAPK kinase was involved in inducing the response of potato \u003cem\u003eStRBOHC\u003c/em\u003e and \u003cem\u003eStRBOHD\u003c/em\u003e genes in response to pathogen signals in potato [\u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e78\u003c/span\u003e], and Asai et al. (2008) also illustrated that the MAPK cascade MEK2-SIPK regulates the oxidative burst resulting from the induction of \u003cem\u003eRBOHB\u003c/em\u003e expression in resistance to \u003cem\u003eP. infestans\u003c/em\u003e and \u003cem\u003eColletotrichum orbiculare\u003c/em\u003e of \u003cem\u003eN. benthamiana\u003c/em\u003e [\u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e79\u003c/span\u003e]. Here, we identified the expression of a RBHO-encoded gene \u003cem\u003eTraesCS1A03G0718100\u003c/em\u003e was up-regulated after \u003cem\u003eF. graminearum\u003c/em\u003e infection in YNM158 (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eJ), as well as some genes encoding MAPK kinase (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eG). Therefore, we hypothesized that MAPK kinase in YNM158 might be involved in inducing RBOH gene response to the resistance against \u003cem\u003eF. graminearum\u003c/em\u003e. However, this puzzle requires further investigations.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusions","content":"\u003cp\u003eFHB is a devastating wheat disease that seriously affects the yield and quality of wheat. Up to now, many laboratories around the world have carried out researches on wheat resistance to FHB. It has been proved by practice that the most economical and effective way to resist the damage of wheat FHB is to mine the genes with high resistance to FHB and breed new varieties resistant to FHB. In this study, a translocation line YNM158 carried 7EL chromosome fragment obtained by distant hybridization not only has excellent resistance to FHB, but also had stable agronomic traits that could potentially be used in FHB resistance breeding. Moreover, transcriptome analysis showed that the 7EL chromosome fragment not only carried the genes that can detoxify DON, but also have the genes that can affect the colonization of \u003cem\u003eF. graminearum\u003c/em\u003e during the early stage of infection. In addition, the introgression of 7EL fragment altered the gene expression and activated the special resistance pathway in YNM158 after \u003cem\u003eF. graminearum\u003c/em\u003e inoculation. It is demonstrated that YNM158 may have a variety of molecular mechanisms to against \u003cem\u003eF. graminearum\u003c/em\u003e infection, and show a high resistance to FHB phenotype. Therefore, these results not only provide a new germplasm for wheat resistance to FHB, but also provide new insights into the molecular mechanism of wheat resistance to FHB, and also provide a new way for breeding new varieties with high resistance to FHB.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003ePlant materials\u003c/h2\u003e \u003cp\u003e \u003cem\u003eTriticum aestivum\u003c/em\u003e cv. Chinese Spring (CS), Su Mai3 (SU3), An Nong8455 (AN8455), and Yang Mai158 (YM158) are maintained at Yangzhou University, China. Wheat cultivar YM158 was pollinated with the \u003csup\u003e60\u003c/sup\u003eCo-γ-irradiated pollen of the long-arm translocation line TW-7EL1 (T7BS\u0026middot;7EL) of 7E chromosome with excellent FHB resistance developed in previous studies. The small fragment of 7EL chromosome translocation line Yangnongmai158 (YNM158) was selected from the hybrid progeny in this study.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eCell cycle synchronization and preparation of mitotic chromosomes\u003c/h2\u003e \u003cp\u003eCell cycle synchronization and slide preparation followed Lei et al. [\u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e80\u003c/span\u003e] with minor modifications. Seeds were soaked in water for 3\u0026ndash;5 hours and germinated on moist filter paper for 2 days in the dark at 25\u0026deg;C. When the roots grew to about 2.5 cm long, the roots were treated with 2 \u0026micro;mol/L amiprophosmethyl (APM) for 2.5 h. Then the root tips were cut off and treated in a nitrous oxide gas chamber for 1 h. After that the root tips were fixed in ice-cold 90% acetic acid for 8 min, washed with sterile distilled water (ddH\u003csub\u003e2\u003c/sub\u003eO) and stored in 70% ethanol at -20\u0026deg;C until use. For slide preparation, root tips were washed in ddH\u003csub\u003e2\u003c/sub\u003eO for 5 min. The apical meristem of roots was cut and incubated in 25 \u0026micro;L of enzyme solution containing 2% cellulase Onozuka R-10 (Yakult Pharmaceutical, Tokyo) and 1% pectolyase Y23 (ICN) for 1 h at 37\u0026deg;C in a water bath. Meristems were separated with a needle in 50 \u0026micro;L of 100% acetic acid and immediately dropped onto microscope slides using a pipette at a height of approximately 10 cm and then placed in a wet box for about 20 min. The number and location of chromosomes were observed and recorded under the phase contrast objective (Nikon 80i, Japan), and the well-prepared slides were stored in a -70\u0026deg;C refrigerator until use.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eGenomic in situ hybridization (GISH) and Fluorescence in situ hybridization (FISH) analysis\u003c/h2\u003e \u003cp\u003eThe techniques of GISH and ND-FISH followed those of Tang et al. [\u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e81\u003c/span\u003e]. Total genomic DNA of \u003cem\u003eTh. elongatum\u003c/em\u003e was labeled with digoxigenin-12-dUTP by Nick Translation method and used as a probe for GISH. The repetitive sequences oligo-pSc119.2 and oligo-pAs1 were synthesized as the probes. And the 5\u0026rsquo; end of oligo-pSc119.2 was labeled by 6-carboxyfluorescein (6-FAM), the 5\u0026rsquo; ends of oligo-pAs1 was labeled by 6-carboxytetramethylrhodamine (Tamra). The labeled probes were dissolved in 2 \u0026times; SSC and 1\u0026times; TE buffer (pH 7.0), dropped to the prepared slides. After that the slides were covered with a coverslip and placed in a humidified hybridization cassette at 37\u0026deg;C for 10 h, and then transferred into 2\u0026times;SSC for 2 min at room temperature. Finally, the slides were quickly dried, and then 6.5 \u0026micro;L DAPI was added to each slide (Vector, No. H-1200). After ND-FISH analysis, the slides were washed in 2\u0026times;SSC for 2 min at room temperature. After drying the same slides were subjected to GISH analysis. Hybridization signals were observed using a fluorescent microscope and images were obtained with a CCD camera (Color Cooled Digital DS-Fi1c, Nikon 80i, Japan).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eEvaluation of Disease Resistance\u003c/h2\u003e \u003cp\u003eThe translocation line YNM158 and its hybrid offspring were screened for FHB resistance in the field and greenhouse in 2021 and 2022. At early flowering stage, the central spikelet was injected into 10 \u0026micro;L fungal suspension (50,000 spores/mL), and at least three spikes from each plant was injected. Following inoculation, the plants were misted for 72 h for FHB development. After 21 days, all infected spikelets per inoculated spike were counted. Wheat cultivars An8455 served as the susceptible controls in both the field and greenhouse, respectively, and SU3 served as the resistant control in both the field and greenhouse.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eDe novo assembly of RNA-seq reads and quantifying gene expression\u003c/h2\u003e \u003cp\u003eThe transcriptome analysis was comprised of 8 time points after inoculation with \u003cem\u003eF. graminearum\u003c/em\u003e: 0 hpi, 0.5 hpi, 2 hpi, 8 hpi, 24 hpi, 48 hpi, 72 hpi and 96 hpi. Three spikes after inoculation was randomly selected for each time were mixed to extract RNA. Total RNA was extracted using Trizol reagent kit (Invitrogen) according to the manufacturer\u0026rsquo;s protocol. RNA quality was assessed on an Agilent 2100 Bioanalyzer (Agilent Technologies) and checked by using RNase-free agarose gel electrophoresis. The mRNA was enriched by Oligo (dT) beads. Then the enriched mRNA was fragmented and used as the template for cDNA synthesis. The cDNA fragments were sequenced using Illumina HiSeq2500 by Gene Denovo Biotechnology Co. For the analysis process after sequencing, referred to the article published by Dai et al. [\u003cspan citationid=\"CR82\" class=\"CitationRef\"\u003e82\u003c/span\u003e]. The genome of Chinese Spring (IWGSC RefSeq v2.1) and the genome of \u003cem\u003eTh. elongatum\u003c/em\u003e (GWHABKY00000000) were used as the reference genome in this study. The obtained sequences were submitted on the sequence read archive data (BioProject ID: PRJNA1011388).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eQuantitative real-time polymerase chain reaction\u003c/h2\u003e \u003cp\u003eThree spikes after inoculation was randomly selected for each time were mixed to extract RNA. qRT-PCR was performed under the following program: 94\u0026deg;C for 5 min, and then 40 cycles: 94\u0026deg;C for 5 min followed by 60\u0026deg;C for 30 s. For the melt curve analysis, the following program was included after 40 cycles: 95\u0026deg;C for 10 s followed by 60\u0026deg;C for 30 s and a constant increase from 60 to 95\u0026deg;C. The relative expression levels were determined using the \u0026shy;2\u003csup\u003e\u0026minus;ΔΔCt\u003c/sup\u003e method. qRT-PCR assays were carried out in three independent biological samples pretreatment and three technical replicates per samples. The primers used for this analysis were shown in Table S19.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eData analysis\u003c/h2\u003e \u003cp\u003eAll data were statistically analyzed using the IBM SPSS Statistics 25 software with pairwise comparisons of LSD to identify differences. The data conforming to normal distribution and homogeneity of variance use one-way ANOVA; otherwise Kruskal-Wallis one-way ANOVA was used. Significant differences (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) were indicated by different letters. The GraphPad Prism 8 software was used to draw figures.\u003c/p\u003e \u003c/div\u003e"},{"header":"Abbreviations","content":"\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eFHB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003eFusarium\u003c/em\u003e head blight\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eDON\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eDeoxynivalenol\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eGST\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eGlutathione S-transferase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eGSH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eGlutathione\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAPM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eAmiprophosmethyl\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eGISH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eGenomic in situ hybridization\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eFISH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eFluorescence in situ hybridization\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eND-FISH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eNon-denaruring fluorescence in situ hybridization\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ehpi\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHours post-inoculation\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eDEG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eDifferentially expressed genes\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePIP5K\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePhosphatidylinositol 4-phosphate 5-kinase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eBIP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eImmunoglobulin-binding protein\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHsp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHeat shock protein\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAPX\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eAscorbate peroxidase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eGR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eGlutathione reductase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHypersensitive response\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eFLS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eFlavonol synthase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCHI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eChalcone isomerase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCHS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eChalcone synthase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHCT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHydroxycinnamoyl-coa shikimate\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eMAPK\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eMitogen-activated protein kinase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eRBOH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eRespiratory burst oxidase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eWGCNA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eWeighted gene co-expression network analysis\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePTI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePathogen-associated molecular pattern-trigged immunity\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePRRs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePattern recognition receptors\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePLKs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eReceptor-like kinases\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePLPs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eReceptor-like proteins\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eROS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eReactive oxygen species\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eMIK2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eMDIS-interacting receptor-like kinase 2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCWIN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eCell wall invertase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePLC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePhospholipase C\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eSalicylic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eJA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eJasmonate\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eRER\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eRough endoplasmic reticulum\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCRT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eCalreticulin\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePDI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eProtein disulfide isomerase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePCR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePolymerase chain reaction\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank the participants for partaking in this study. The authors would like to thank the reviewers whose constructive comments are very helpful for strengthening the presentation of this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eY.D., data curation, data analysis, visualization, methodology, writing-original draft. W.F., data analysis, material identification. S.C., data curation, investigation of agronomic traits, material planting. J.S., data analysis, material identification, evaluation of fhb resistance. H.L., material planting, material identification. J.L., investigation of agronomic traits, evaluation of FHB resistance. H.M. (Haigang Ma), evaluation of FHB resistance. Y.W., formal analysis, validation. Y.G., visualization. J.Z., and B.W., material radiation. J.C, conceptualization, supervision, data curation. H.M. (Hongxiang Ma), conceptualization, supervision, data curation, funding acquisition, project administration, resources, writing-review \u0026amp; editing.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was funded by the Project of Zhongshan Biological Breeding (ZSBBL-KY2023-02-3, BM2022008-02), Seed Industry Revitalization Project of Jiangsu Province (JBGS2021047), Jiangsu Key Project for the Research and Development (BE2022346), and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe genome sequences of Chinese Spring were downloaded from the website: http://www.wheatgenome.org/News2/IWGSC-RefSeq-v2.0-now-available-at-URGI. The genome sequences of \u003cem\u003eTh. elongatum\u003c/em\u003e were downloaded from the website: https://ngdc.cncb.ac.cn/gwh/Assembly/965/show. The transcriptome sequencing data generated in this study have been deposited in the NCBI Sequence Read Archive (SRA) database under accession number PRJNA1011388. And the datasets generated or analyzed during this study are included in this article and its additional file or are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eCuthbert PA, Somers DJ, Thomas J, Cloutier S, Brule-Babel A. Fine mapping \u003cem\u003eFhb1\u003c/em\u003e, a major gene controlling \u003cem\u003eFusarium\u003c/em\u003e head blight resistance in bread wheat (\u003cem\u003eTriticum aestivum\u003c/em\u003e L.). 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Agronomy J. 2021;113:3639-3652.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Fusarium head blight, wheat-Th. elongatum translocation line, transcriptome analysis, disease resistance pathway","lastPublishedDoi":"10.21203/rs.3.rs-4079736/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4079736/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eFusarium \u003c/em\u003ehead blight (FHB) caused by \u003cem\u003eFusarium graminearum\u003c/em\u003e species complex is a destructive disease in wheat worldwide. Lack of FHB resistant germplasm is a barrier in wheat breeding for the resistance to FHB. \u003cem\u003eThinopyrum elongatum\u003c/em\u003e is an important relative species successfully used for genetic improvement in wheat.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn this study, a translocation line YNM158 with a YM158 genetic background and carrying the fragment of diploid \u003cem\u003eTh. elongatum\u003c/em\u003e 7EL chromosome created by \u003csup\u003e60\u003c/sup\u003eCo-γ radiation showed high resistance to FHB under both filed and greenhouse conditions. The transcriptome analysis validated that the horizontal transfer gene \u003cem\u003eGST\u003c/em\u003e is one of the important contributors to FHB resistance in pathogen infection stage, whereas 7EL chromosome fragment also carries other genes regulated by \u003cem\u003eF. graminearum\u003c/em\u003e during the colonization stage. In addition, the introgression of 7EL fragment affected the expression of wheat genes which were enriched in the resistance pathways including phosphatidylinositol signaling system, protein processing in endoplasmic reticulum, plant-pathogen interaction and MAPK signaling pathway at different stages after \u003cem\u003eF. graminearium\u003c/em\u003e infection.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study provides a novel germplasm for wheat resistance to FHB and new insights into the molecular mechanism of wheat resistance to FHB.\u003c/p\u003e","manuscriptTitle":"Transcriptome analysis revealed the potential mechanism of a wheat-Th. elongatum translocation line YNM158 against Fusarium head blight","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-27 12:11:47","doi":"10.21203/rs.3.rs-4079736/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"1e4e645a-082d-4e73-b6b3-9fb450f04a9e","owner":[],"postedDate":"March 27th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-05-02T20:55:29+00:00","versionOfRecord":[],"versionCreatedAt":"2024-03-27 12:11:47","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4079736","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4079736","identity":"rs-4079736","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}