Search

Key Message: We revealed the neglected genetic relationships of resistance for six major wheat diseases and established a haploblock-based catalogue with novel forms of resistance by multi-trait haplotype characterisation. Genetic potential to improve multiple disease resistance was highlighted through haplotype stacking simulations. Wheat production is threatened by numerous fungal diseases, but the potential to breed for multiple disease resistance (MDR) mechanisms is yet to be explored. Here, significant global genetic correlations and underlying local genomic regions were identified in the Vavilov wheat diversity panel for six major fungal diseases, including biotrophic leaf rust (LR), yellow rust (YR), stem rust (SR), hemibiotrophic crown rot (CR), and necrotrophic tan spot (TS) and Septoria nodorum blotch (SNB). By adopting haplotype-based local genomic estimated breeding values, derived from an integrated set of 34,899 SNP and DArT markers, we established a novel haplotype catalogue for resistance to the six diseases in over 20 field experiments across Australia and Ethiopia. Haploblocks with high variances of haplotype effects in all environments were identified for three rusts, and pleiotropic haploblocks were identified for at least two diseases, with four haploblocks affecting all six diseases. Through simulation, we demonstrated that stacking optimal haplotypes for one disease could improve resistance substantially, but indirectly affected resistance for other five diseases, which varied depending on the genetic correlation with the non-target disease trait. On the other hand, our simulation results combining beneficial haplotypes for all diseases increased resistance to LR, YR, SR, CR, TS, and SNB, by up to 48.1%, 35.2%, 29.1%, 12.8%, 18.8%, and 32.8%, respectively. Overall, our results highlight the genetic potential to improve MDR in wheat. The haploblock-based catalogue with novel forms of resistance provides a useful resource to guide desirable haplotype stacking for breeding future wheat cultivars with MDR., Competing Interests: Declarations. Conflict of interest: All authors declare that they have no conflicts of interest., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

NLR receptor is suggested as a component of plant nonhost resistance (NHR). However, the evolutionary process of how plants develop receptors for recognizing broad-spectrum pathogens is still elusive. Here, we observe that multiple RxLR effector families including 12 reported avirulence effectors of Phytophthora infestans are broadly conserved across the Phytophthora species. We select 69 effectors distributed into 8 families from 6 Phytophthora species, and confirm that 60.87% of the tested effectors are recognized by Solanum NLRs according to their defined families. Furthermore, we confirm that expression of R1, R8, and Rpi-amr1 confer broad-spectrum resistance against multiple Phytophthora species. Combined results suggest that conserved effector families of Phytophthora species allow solanaceous plants to recognize broad-spectrum pathogens via NLRs that originally reported to recognize P. infestans. Thus, NLR-mediated recognition would contribute to NHR against pathogens that possess similar repertoires of effectors. Moreover, this homology-based approach would be applicable to other plant-pathogen systems and provide an alternative strategy of genetic mapping to identify functional NLRs against various crop-threatening pathogens., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

The study was designed to validate the previously reported 34 SSR markers using 78 chilli genotypes to detect significant trait specific markers as well as superior genotypes resistant to Phytophthora capsici root rot (PcRR). In this context, the identification of germplasm with higher yield per plant (YPP) leads to hype in stress tolerance index (STI) in genotypes, Chakwal3 (11.98), Greenfire (10.14), Advanta5017 (9.94) and Chakwal4 (7.8). The identified genotypes were also found as resistant and moderately resistant due to existence of below 50% of disease incidence. Moreover, biplot showed the interrelation of STI with YPP through the formation of acute angle by their respective vectors. In the current study, the markers Hpms1172 and CAMS177 was found significant for STI. However, the marker CAMS066 was found associated with relative cell injury, CA06g27450 with disease incidence and CAMS173 with relative leaf damage. The bright bands on gel pictures of significant markers showed the association of these markers with resistant genotypes i.e. Chakwal3, Advanta-5017 and Chakwal4 as well as with a single moderately resistant genotype i.e. Greenfire. The markerstudes confirmed the phenotypic data by showing association of markers i.e. Hpms1172 and CAMS177, r with stress tolerance index. The principal coordinate analysis aligned with the results obtained from marker-assisted selection. Thus, currently practiced marker assisted selection detected high yielding genotypes in PcRR disease stress condition that will be helpful in progressing breeding programs in chilli., Competing Interests: Declarations Ethics approval and consent to participate This study does not include human or animal subjects. Consent for publication Not applicable. Competing interests The authors stated that they had no interest which might be perceived asposing a conflict or bias. Statement on guidelines All experimental studies and experimental materials involved in this research are in full compliance with relevant institutional, national and international guidelines and legislation., (© 2024. The Author(s).)

Bacterial stem and root rot (BSRR) in sweetpotato caused by Dickeya dadantii is one of the ten major diseases of sweetpotatoes in China. However, the molecular mechanism underlying the resistance of sweetpotato to D. dadantii remains unclear. This study adopted a resistance identification assay that conformed Guangshu87 (GS87) as BSRR-resistant and Xinxiang (XX) as susceptible. Compared to XX, GS87 effectively prevented the invasion and dissemination of D. dadantii in planta. An RNA sequencing (RNA-seq) analysis identified 54,844 expressed unigenes between GS87 and XX at four different stages. Further, it revealed that GS87 was more able to regulate the expressions of more unigenes after the inoculation with D. dadantii, including resistance (R) and transcription factors (TF) genes. Moreover, content measurements of disease resistance-related phytohormones showed that both jasmonic acids (JAs) and salicylic acids (SAs) accumulated in D. dadantii-inoculated sweetpotatoes, and JAs may negatively regulate sweetpotato resistance against D. dadantii and accumulated faster than SAs. Meanwhile, determinations of ROS production rate and relevant enzymatic/non-enzymatic activity highlighted the vital roles of reactive oxygen species (ROS) and superoxide dismutase (SOD) in confering GS87 resistance against D. dadantii. Additionally, several hub genes with high connectivity were highlighted through Protein-Protein interaction (PPI) network analysis. In summary, the findings in this study contribute to the understanding of the different responses of resistant and susceptible sweetpotato cultivars to D. dadantii infection, and it also provide the first insight into the relevant candidate genes and phytohormones involved in the resistance of sweetpotato to D. dadantii., Competing Interests: Declarations Ethics approval and consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Nucleotide-binding leucine-rich repeat (NLR) disease resistance genes typically confer resistance against races of a single pathogen. Here, we report that Yr87/Lr85, an NLR gene from Aegilops sharonensis and Aegilops longissima, confers resistance against both P. striiformis tritici (Pst) and Puccinia triticina (Pt) that cause stripe and leaf rust, respectively. Yr87/Lr85 confers resistance against Pst and Pt in wheat introgression as well as transgenic lines. Comparative analysis of Yr87/Lr85 and the cloned Triticeae NLR disease resistance genes shows that Yr87/Lr85 contains two distinct LRR domains and that the gene is only found in Ae. sharonensis and Ae. longissima. Allele mining and phylogenetic analysis indicate multiple events of Yr87/Lr85 gene flow between the two species and presence/absence variation explaining the majority of resistance to wheat leaf rust in both species. The confinement of Yr87/Lr85 to Ae. sharonensis and Ae. longissima and the resistance in wheat against Pst and Pt highlight the potential of these species as valuable sources of disease resistance genes for wheat improvement., Competing Interests: Competing interests The authors declare the following competing interests: D.S., R.A., R.K, E.M., A.M-D., and A.S. are inventors on the US patent application 63/250,413 filed by Ramot (Tel Aviv University) and relating to the use of Yr87/Lr85 for leaf and stripe rust resistance in transgenic wheat. All other authors claim no competing interests., (© 2024. The Author(s).)

Background: Sooty mold (SM), caused by Cladosporium species, is a pervasive threat to tea plant health, affecting both canopy structure and crop yield. Despite its significance, understanding the complex interplay between defense genes and metabolites in tea plants across various SM-infected canopy layers remains limited. Our study employed hyperspectral imaging, transcriptomic profiling, and metabolomic analysis to decipher the intricate mechanisms underlying the tea plant's response to SM infection., Results: Our hyperspectral imaging identified three critical wavelengths (552, 673, and 800 nm) inflection points associated with varying degrees of SM infection. This non-invasive method allows for the precise assessment of disease progression. Concurrently, transcriptome analysis revealed a wealth of differentially expressed genes (DEGs) enriched in metabolic pathways, secondary metabolite biosynthesis, and plant-pathogen interactions. Cluster analysis highlighted an intensified immune response in A2 and A3 samples. A comprehensive metabolomic profile identified 733 co-changed metabolites in SM-infected leaves, with alcohols, lipids (free fatty acids), hydrocarbons, and amino acids significantly accumulating in A1, while flavonoids were predominantly upregulated in A2 and A3. Weighted Gene Co-Expression Network Analysis (WGCNA) uncovered five hub genes (Dormancy-associated protein, Serine/threonine-protein phosphatase, ABC transporter, and some uncharacterized proteins) and two hub metabolites (D-Mannitol and 17-Hydroxylinolenic Acid) that exhibit significant relationships with DEGs and metabolites. Further co-expression analysis indicated that tea plants mainly employed genes and metabolites related to the biosynthesis of secondary metabolites, plant hormone signal transduction, and plant-pathogen interaction to combat SM., Conclusion: This study establishes a foundation for understanding the immune mechanisms of tea plants across different canopy layers in response to SM infection. It not only sheds light on the complex defense strategies employed by tea plants but also identifies candidate genes and metabolites crucial for enhancing tea plant breeding and resistance to SM., Competing Interests: Declarations Ethics approval and consent to participate Not applicable. All methods were carried out according to relevant guidelines and regulations. Consent to publish Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Black shank disease is the main disease affecting tobacco crops worldwide, and the main impacted by the disease are the stem base and root. At present, transgenic technology is an effective method to improve plant disease resistance through transgenic technology. In this study, the EuCHIT73.88 gene was cloned from Eucommia ulmoides Oliver (E. ulmoides) by using RT-PCR. The full length of the gene was 897 bp, encoding 298 amino acid residues. An overexpression vector of from the EuCHIT73.88 gene driven by the 35S promoter was constructed and transferred into tobacco plants via transgenic technology. After inoculation with the black shank pathogen, the number of visible lesions on the stems and leaves of the transgenic tobacco variety EuCHIT73.88 was significantly shorter than that on the stems and leaves of the of wild type (WT) and empty vector (EV) plants, and the lesion area was significantly smaller than on the stems and leaves of the WT and EV plants. With increasing inoculation time, introduction of the WT and EV vectors was obviously lethal, whereas transgenic tobacco only exhibited wilted characteristics, and the stems were black, which indicated that the EuCHIT73.88 gene could improve the resistance of tobacco to black shank disease. Furthermore, the activity of protective enzymes and the gene expression of resistance-related proteins were measured. The results showed that compared with those of the WT and EV plants, the CAT and POD activities of the TP tobacco plants were greater, peaking at 72 h at concentrations of 446.87 U/g and 4562.24 U/g, which were 1.63 and 1.61 times greater than those of the WT and EV plants, respectively. This indicated that CAT and POD may be involved in the process of disease resistance of in the transgenic plants. The MDA content of the transgenic tobacco plants was significantly lower than that of the WT and EV plants with increasing EuCHIT73.88 expression, thus indicating that the overexpression of the transgenic EuCHIT73.88 gene could alleviate the levels of lipid peroxidation and reduce the damage to plant cell membranes. The expression of disease-related protein genes (PR2, PR5, PR1a, PDF1.2 and MLP423) was significantly greater in the EuCHIT73.88 ransgenic tobacco than in the WT and EV-transgenic tobacco. and these findings consistently showed that EuCHIT73.88 could improve the resistance to black shank., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Plasmodiophora brassicae, the causative agent of clubroot disease, establishes a long-lasting parasitic relationship with its host by inducing the expression of sugar transporters. Previous studies have indicated that most BrSWEET genes in Chinese cabbage are up-regulated upon infection with P. brassicae. However, the key BrSWEET genes responsive to P. brassicae have not been definitively identified. In this study, we selected five BrSWEET genes and conducted a functional analysis of them. These five BrSWEET genes showed a notable up-regulation in roots after P. brassicae inoculation. Furthermore, these BrSWEET proteins were localized to the plasma membrane. Yeast functional complementation assays confirmed transport activity for glucose, fructose, or sucrose in four BrSWEETs, with the exception of BrSWEET2a. Mutants and silenced plants of BrSWEET1a, -11a, and -12a showed lower clubroot disease severity compared to wild-type plants, while gain-of-function Arabidopsis thaliana plants overexpressing these three BrSWEET genes exhibited significantly higher disease incidence and severity. Our findings suggested that BrSWEET1a, BrSWEET11a, and BrSWEET12a play pivotal roles in P. brassicae-induced gall formation, shedding light on the role of sugar transporters in host-pathogen interactions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

5-Azacytidine (AZ) is a DNA methylation inhibitor that has recently demonstrated potential in regulating fruit quality through exogenous application. In this study, we treated mandarin fruits for 4-day storage. Noteworthy were the induced degreening and the enhanced citrus aroma of fruits under AZ treatment, involving the promotion of chlorophyll degradation, carotenoid biosynthesis, and limonene biosynthesis. Key genes associated with these processes exhibited expression level increases of up to 123.8 times. Additionally, AZ treatment activated defense-related enzymes and altered phenylpropanoid carbon allocation towards lignin biosynthesis instead of flavonoid biosynthesis. The expression levels of lignin biosynthesis-related genes increased by nearly 100 times, leading to fortified lignin that is crucial for citrus defense against Penicillium italicum. Currently, the underlying mechanisms of such intense AZ-induced changes in gene expressions remain unclear and further research could help establish AZ treatment as a viable strategy for citrus preservation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

MYC2 is a class of bHLH family transcription factors and a major regulatory factor in the JA signaling pathway, and its molecular function in tobacco has not been reported. In this study, CRISPR/Cas9-mediated MYC2 gene NtMYC2a knockout mutants at tobacco was obtained and its agronomic traits, disease resistance, and chemical composition were identified. Comparing with the WT, the leaf width of the KO-NtMYC2a was narrowed, the nornicotine content and mecamylamine content increased significantly and the resistance to Ralstonia solanacearum significantly decreased. The transcriptome sequencing results showed that DEGs related to immunity, signal transduction and growth and development were enriched between KO-NtMYC2a and WT. NtJAR1 and NtCOI1 in KO-NtMYC2a were down-regulated to regulating the JA signaling pathway, result in a significant decrease in tobacco's resistance to R. solanacearum. Our research provides theoretical support for the functional research of MYC2 and the study of the mechanism of tobacco bacterial wilt resistance., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Key Message: Robust QTLs conferring resistance to bacterial leaf streak in wheat were mapped on chromosomes 3B and 5A from the variety Boost and on chromosome 7D from the synthetic wheat line W-7984. Bacterial leaf streak (BLS), caused by Xanthomonas translucens pv. undulosa poses a significant threat to global wheat production. High levels of BLS resistance are rare in hexaploid wheat. Here, we screened 101 diverse wheat genotypes under greenhouse conditions to identify new sources of BLS resistance. Five lines showed good levels of resistance including the wheat variety Boost and the synthetic hexaploid wheat line W-7984. Recombinant inbred populations derived from the cross of Boost × ND830 (BoostND population) and W-7984 × Opata 85 (ITMI population) were subsequently evaluated in greenhouse and field experiments to investigate the genetic basis of resistance. QTLs on chromosomes 3B, 5A, and 5B were identified in the BoostND population. The 3B and 5A QTLs were significant in all environments, but the 3B QTL was the strongest under greenhouse conditions explaining 38% of the phenotypic variation, and the 5A QTL was the most significant in the field explaining up to 29% of the variation. In the ITMI population, a QTL on chromosome 7D explained as much as 46% of the phenotypic variation in the greenhouse and 18% in the field. BLS severity in both populations was negatively correlated with days to heading, and some QTLs for these traits overlapped, which explained the tendency of later maturing lines to have relatively higher levels of BLS resistance. Markers associated with the QTLs were converted to KASP markers, which will aid in the deployment of the QTLs into elite lines for the development of BLS-resistant wheat varieties., Competing Interests: Declarations Conflict of interest The authors have no relevant financial or non-financial interests to disclose. Ethics approval NA. Consent to participate NA. Consent to publish NA., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Key Message: Using QTL mapping and GWAS, two candidate genes (Zm00001d051039 and Zm00001d051147) were consistently identified across the three different environments and BLUP values. GWAS analysis identified the candidate gene, Zm00001d044845. These genes were subsequently validated to exhibit a significant association with maize gray leaf spot (GLS) resistance. Gray leaf spot (GLS) is a major foliar disease of maize (Zea mays L.) that causes significant yield losses worldwide. Understanding the genetic mechanisms underlying gray leaf spot resistance is crucial for breeding high-yielding and disease-resistant varieties. In this study, eight tropical and subtropical germplasms were crossed with the temperate germplasm Ye107 to develop a nested association mapping (NAM) population comprising 1,653 F2:8 RILs, consisting of eight recombinant inbred line (RIL) subpopulations, using the single-seed descent method. The NAM population was evaluated for GLS resistance in three different environments, and genotyping by sequencing of the NAM population generated 593,719 high-quality single-nucleotide polymorphisms (SNPs). Linkage analysis and genome-wide association studies (GWASs) were conducted to identify candidate genes regulating GLS resistance in maize. Both analyses identified 25 QTLs and 149 SNPs that were significantly associated with GLS resistance. Candidate genes were screened 20 Kb upstream and downstream of the significant SNPs, and three novel candidate genes (Zm00001d051039, Zm00001d051147, and Zm00001d044845) were identified. Zm00001d051039 and Zm00001d051147 were located on chromosome 4 and co-localized in both linkage (qGLS4-1 and qGLS4-2) and GWAS analyses. SNP-138,153,206 was located 0.499 kb downstream of the candidate gene Zm00001d051039, which encodes the protein IN2-1 homolog B, a homolog of glutathione S-transferase (GST). GSTs and protein IN2-1 homolog B scavenge reactive oxygen species under various stress conditions, and GSTs are believed to protect plants from a wide range of biotic and abiotic stresses by detoxifying reactive electrophilic compounds. Zm00001d051147 encodes a probable beta-1,4-xylosyltransferase involved in the biosynthesis of xylan in the cell wall, enhancing resistance. SNP-145,813,215 was located 2.69 kb downstream of the candidate gene. SNP-5,043,412 was consistently identified in three different environments and BLUP values and was located 8.788 kb downstream of the candidate gene Zm00001d044845 on chromosome 9. Zm00001d044845 encodes the U-box domain-containing protein 4 (PUB4), which is involved in regulating plant immunity. qRT-PCR analysis showed that the relative expression levels of the three candidate genes were significantly upregulated in the leaves of the TML139 (resistant) parent, indicating that these three candidate genes could be associated with resistance to GLS. The findings of this study are significant for marker-assisted breeding aimed at enhancing resistance to GLS in maize and lay the foundation for further elucidation of the genetic mechanisms underlying resistance to gray leaf spot in maize and breeding of new disease-resistant varieties., Competing Interests: Declarations Conflict of interest The authors declare that they have no competing interests., (© 2024. The Author(s).)

Small molecules targeting G-quadruplexes (G4s) in viruses could inhibit viral proliferation. The 1a protein of cucumber mosaic virus (CMV) act as RNA-dependent RNA polymerase (RdRp) that plays a crucial role in regulating the replication of CMV. In this study, four putative G4 sequences (CMV PQS1-PQS4) in the genetic coding region of CMV 1a were identified, and three of them (PQS2, PQS3, and PQS4) were confirmed to fold into G4 structures. The G4-ligand, RHPS4, could bind to CMV PQS2 and PQS4 with a strong binding affinity and preferred to interact with the 3' terminal G-quartet surfaces of CMV PQS2, and 5' terminal of CMV PQS4. RHPS4 was also found to stabilize the CMV PQS2 and PQS4 G4s. Further studies revealed that RHPS4 exhibited an excellent anti-CMV activity. This study suggested that CMV PQS2 and PQS4 could be considered potential targets for screening viral inhibitors.

Direct immersion solid-phase microextraction coupled with gas chromatography-mass spectroscopy was used to create chemical fingerprints of annual ryegrass cultivars ( Lolium rigidum ). Extracts made of the inflorescences of four cultivars and one accession of annual ryegrass were assessed to identify differential metabolites between those resistant to and susceptible to bacterial galls associated with annual ryegrass toxicity (ARGT). Numerous compounds were identified. Principal component analysis showed distinct clustering of metabolites from disease-resistant and disease-susceptible cultivars. Partial least-squares-discriminant analysis identified sterols, esters, aldehydes, and terpenes that correlated with resistance to galls formation. Esters, sterols, phenols, heterocyclics, fatty acids, organofluorides, and siloxanes were predominant in resistant genotypes, whereas alcohols, aldehydes, terpenes, and hydrocarbons were predominant in susceptible genotypes. The identification of differentially expressed metabolites provides potential chemical markers to guide breeding strategies for ARGT resistance in ryegrass.

Objective: Cucumber mosaic virus (CMV) is a significant pathogen causing quality loss in spinach. Although host genetic resistance is the primary method of managing CMV infection in this crop, CMV resistance genes are not widely utilized in spinach breeding programs as the genetic and molecular mechanisms underlying resistance are not yet fully understood. CMV infections were therefore studied in different lines of spinach plants, and their progeny, to develop a model of the genetic basis of CMV resistance., Results: Visual observations and RT-PCR assays revealed that three monoecious lines (03-258, 03-263, and 03-336) were susceptible to CMV, while three traditional resistant cultivars and a near-isogenic line (NIL-M) exhibited resistance. A dioecious line (03-009) consisted of susceptible and resistant plants. Notably, resistant plants did not exhibit the lesions typical of the hypersensitive response. Genetic analysis of progeny from the cross NIL-M × 03-336 indicated that a single dominant allele (designated SRCm1, standing for Spinach Resistance to CMV 1) controlled CMV resistance; analysis of sib-cross progeny populations derived from line 03-009 supported this conclusion. These results offer a valuable model for CMV resistance in spinach and will enhance future breeding programs., Competing Interests: Declarations Ethics approval and consent to participate Not applicable for this study. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Background: Bigleaf hydrangea (Hydrangea macrophylla) is a widely cultivated ornamental plant species. Leaf spot disease, caused by Corynespora cassiicola, poses a significant threat to the ornamental quality and economic value of hydrangeas. However, the disease resistance breeding of hydrangea is limited due to the lacking of resistant varieties and genes., Results: This study evaluated ten hydrangea varieties for their resistance to leaf spot disease. Among them, 'White Angel' and 'Ocean Heart' were screened out as representative varieties for resistance and susceptibility, respectively, on the basis of evaluation. Physiological and biochemical indices, phytohormones, and transcriptomic changes were measured in the leaves of both varieties at 0 and 24 h post inoculation with C. cassiicola. The results showed that C. cassiicola infection significantly increased abscisic acid (ABA) contents in both varieties; however, the increase was significantly higher in the susceptible variety 'Ocean Heart' compared to the resistant variety 'White Angel' (p < 0.05). Moreover, exogenous ABA (100 µM) decreased the leaves' resistance to C. cassiicola of both varieties, underscoring its key role in reduced disease resistance. Transcriptome profiling revealed 17,087 differentially expressed genes (DEGs) responding to C. cassiicola between the two varieties. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated significant enrichment of DEGs in "Plant hormone signal transduction", particularly related to ABA signaling (HmPP2C and HmABFs). In addition, the expression of ABA biosynthesis genes (HmZEP3, HmABA2, and HmAAO3) was upregulated in both varieties. Meanwhile, the ABA catabolism gene (HmCYP707A4) exhibited significantly upregulated expression in the resistant variety 'White Angel' and downregulated expression in the susceptible variety 'Ocean Heart'. Intriguingly, the expression of HmCYP707A4 was 15-fold higher in 'White Angel' than in 'Ocean Heart'., Conclusion: In summary, these findings highlight the crucial role of ABA in the resistance of bigleaf hydrangea to leaf spot disease and provide valuable genetic resources for breeding programs to enhance the disease resistance in hydrangeas., Competing Interests: Declarations Ethics approval and consent to participate Not applicable. Our research did not involve any human or animal subjects, material, or data. We declare that the plant material in the experiment was collected and studied in accordance with relevant institutional, national, and international guidelines and legislation. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Plant pathogens secrete nuclear effectors into the host nuclei to modulate the host immune system. Although several nuclear effectors of fungal pathogens have been recently reported, the molecular mechanism of NLS-associated transport vehicles of nuclear effectors and the roles of NLS in transcriptional reprogramming of host immunity genes remain enigmatic. We previously reported the MoHTR1, a nuclear effector of the rice blast fungus, Magnaporthe oryzae. MoHTR1 is translocated to rice nuclei but not in fungal nuclei. Here, we identify the core NLS (RxKK) responsible for MoHTR1's nuclear localization. MoHTR1 is translocated in the host nucleus through interaction with rice importin α. MoHTR1 NLS empowers it to translocate the cytoplasmic effectors of M. oryzae into rice nuclei. Furthermore, other nuclear effector candidates of the blast pathogen and rice proteins which have RxKK also exhibit nuclear localization, highlighting the crucial role of RxKK in this process. We also unveil the importance of SUMOylation in the stability of MoHTR1 and translocation of MoHTR1 to host nuclei. Moreover, MoHTR1 NLS is essential for the pathogenicity of M. oryzae by reprogramming immunity-associated genes in the host. Our findings provide insights into the significance of plant-specific NLS on fungal nuclear effectors and its role in plant-pathogen interactions., Competing Interests: Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Key Message: The pathogenesis-related 1 gene of watermelon responds to the infection of ZYMV and contributes to the resistance of its host. Zucchini yellow mosaic virus (ZYMV; family Potyviridae) is a single-stranded positive-sense RNA virus that is a serious threat to cucurbits. Previously, we observed a hypersensitivity response (HR) in the systemic leaves of the 938-16-B watermelon line infected with ZYMV, distinct from the typical HR at infected sites. In this study, we confirmed that ZYMV accumulation in 938-16-B was significantly lower than in the susceptible line H1. Upon inoculation, the entry of ZYMV-eGFP into mesophyll cells is restricted into necrotic spots in leaves, indicating that resistance to ZYMV in 938-16-B is linked to the HR. Further, grafting experiments between 938-16-B and susceptible varieties were performed, and revealed an HR induction in susceptible varieties, suggesting the transfer of resistance signal(s) from 938-16-B to susceptible varieties. Through RNA-sequencing and proteomics analyses of the H1 scions on 938-16-B rootstock, a pathogenesis-related 1 (ClPR1) gene was identified. Specifically, ClPR1 expression is unique to ZYMV-infected 938-16-B. Repression of the expression of ClPR1 prevents an HR in 938-16-B. Conversely, overexpression of ClPR1 in susceptible varieties significantly reduces ZYMV accumulation, but an HR was not induced in susceptible line. Besides the virus, jasmonic acid (JA) can also trigger an HR in 938-16-B. Intriguingly, the expression of ClPR1 (Cla97C02G034020) is induced in both of 938-16-B and H1 by MeJA, rather than salicylic acid. These results suggest that HR is associated with the expression of ClPR1 and contributes to resistance to ZYMV in 938-16-B., Competing Interests: Declarations Conflict of interest The authors declare that there are no competing interests associated with this manuscript., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Developing wheat cultivars with durable resistance to powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is crucial for sustainable agriculture. The wheat genotype MYC exhibited high resistance to the Bgt isolate E09 at the seedling stage. Genetic analysis identified a recessive gene, temporarily named PmMYC, responsible for this resistance. Understanding the molecular mechanisms underlying this resistance is essential for advancing breeding programs., Results: Bulked Segregant RNA-Seq revealed numerous alternative splicing events generated following Bgt infection, suggesting powdery mildew may disrupt alternative splicing and affect immune responses. Gene Ontology (GO) analysis indicated significant enrichment of differentially expressed genes in "response to stimuli" and "immune system processes", implying their roles in disease defense. BSR-Seq analysis identified two high-confidence candidate regions for PmMYC on chromosome 2B, spanning 40,451,950 - 102,426,703 bp and 421,707,046-449,840,516 bp. Within these intervals, 740 genes were identified, with nonsynonymous mutations in 46 genes in the parents and bulks. Real-time PCR showed distinct expression profiles in four genes in resistant MYC compared to susceptible Yannong 21. KEGG and COG analyses of differentially expressed genes in candidate intervals revealed enrichment in immune processes related to plant-pathogen interactions, confirming that PmMYC initiated a broad immune response to prevent Bgt invasion., Conclusion: The study identified key genetic intervals and genes involved in the resistance of wheat genotype MYC to Bgt. The identified genes, particularly those with altered expression profiles, could serve as valuable targets for breeding programs aimed at developing wheat cultivars with durable resistance to powdery mildew. These findings enhanced our understanding of plant-pathogen interactions and provided a foundation for future genetic and functional studies., Competing Interests: Declarations Ethics approval and consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Arbuscular mycorrhizal fungi (AMF) serve as both plant symbionts and allies in resisting pathogens and environmental stresses. Mycorrhizal colonization of plant roots can influence the outcomes of plant-pathogen interactions by enhancing specific host defense mechanisms. The transcriptional responses induced by AMF in virus-infected plants remain largely unexplored. In the presented study, we employed a comprehensive transcriptomic approach and qPCR to investigate the molecular determinants underlying the interaction between AMF and potato virus Y (PVY) in Solanum tuberosum L. Our primary goal was to identify the symbiosis- and defense-related determinants activated in mycorrhizal potatoes facing PVY. Through a comparative analysis of mRNA transcriptomes in experimental treatments comprising healthy and PVY-infected potatoes colonized by two AMF species, Rhizophagus regularis or Funneliformis mosseae, we unveiled the overexpression of genes associated with mycorrhiza, including nutrient exchange, lipid transfer, and cell wall remodeling. Furthermore, we identified several differentially expressed genes upregulated in all mycorrhizal treatments that encoded pathogenesis-related proteins involved in plant immune responses, thus verifying the bioprotective role of AMF. We investigated the relationship between mycorrhiza levels and PVY levels in potato leaves and roots. We found accumulation of the virus in the leaves of mycorrhizal plants, but our studies additionally showed a reduced PVY content in potato roots colonized by AMF, which has not been previously demonstrated. Furthermore, we observed that a virus-dependent reduction in nutrient exchange could occur in mycorrhizal roots in the presence of PVY. These findings provide an insights into the interplay between virus and AMF., Competing Interests: Declarations Competing interests The authors have no financial or non-financial interests to disclose., (© 2024. The Author(s).)

Introduction: Rice kernel smut, caused by Tilletia horrida, is becoming an increasingly serious disease in hybrid rice planting, leading to production losses and quality decline of male-sterile rice varieties. Successful infection requires an efficient energy source that the pathogen obtains from rice plants, such as carbohydrates. Glycoside hydrolases (GHs), one of the largest sub-families in the cell wall-degrading enzyme family, play a key role in the infection progress of pathogens. To investigate their roles in facilitating infection, in this study, we identified and characterized genes encoding GH family proteins of T. horrida and further explored the functions and structures of these genes., Materials and Methods: Through genome-wide sequencing and bioinformatics analyses, 52 GH genes were identified from T. horrida, named ThGhd&#95;1 to ThGhd&#95;52. The subcellular location, conserved motifs, and structures of ThGhds were identified by bioinformatics analyses., Results: Phylogenetic analysis revealed that ThGhds with similar domains clustered together, although some proteins clustered in different branches, which might reflect functional diversity. Protein-protein interaction network analysis revealed that ThGhds interact with partner proteins involved in reactive oxygen species signaling, protein kinase activity, and plant hormone signal transduction pathways. RNA-sequencing analysis showed that the expression of ThGhd genes responded differently at different infection time points, with dynamic changes detected during the T. horrida infection process, indicating that these genes are involved in interactions with rice and have potential roles in pathogenic mechanisms., Conclusions: The results of this study provide valuable resources for the structure elucidation of GH family proteins of T. horrida and can help to further elucidate their roles in pathogenesis., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Rhizomania is the most destructive sugar beet disease in the world, and in recent years, it has widespread in most of the sugar beet growing areas in Iran. Since the control of this soil-borne disease is a difficult task, the use of resistant genotypes is known as the best measure against the disease. The ultimate goal of sugar beet breeders is to produce genotypes that can be used in both infected and non-infected fields without any reduction in terms of yield and quality. Twenty-one sugar beet genotypes along with four controls were evaluated in randomized complete block design with four replications in fields with natural infection to rhizomania in five research stations. Important sugar beet traits including root yield, sugar yield, sugar content, and white sugar yield were evaluated for two years (2021 and 2022). For all traits, location was the main source of variation that spanned 33 to 55% of the total sum of the square followed by the location×year×genotype accounted for 3-40% of the variation. Based on the results of analysis of variance, multivariate stability parameters were computed to evaluate the genotypes' stability. The first two principal components (IPCA1 and IPCA2) generated by GGE biplot contributed for 31.3 and 17.5% difference in genotype×environment interaction for root yield, respectively. According to the GGE biplot, genotypes RM-11 and RM-12 were identified as the winning genotypes across environments for both root yield and white sugar yield traits whereas AMMI model identified RM-14 and RM-9 (for root yield) and RM-1 (for white sugar yield) as best genotypes. Based on the ideal genotype ranking, RM-11 and RM-10 were the best performer with a high mean yield as well as stability in the studied environments. The biplot rendered using the weighted average of absolute scores (WAASB) and root yield and white sugar yield identified RM-11 and RM-9 as superior genotypes in terms of yield and stability. The selected genotypes can be used in breeding programs to transfer the disease resistance and cultivar development., (© 2024. The Author(s).)

Storage rots are a significant cause of postharvest losses for the sugar beet crop, however, intrinsic physiological and genetic factors that determine the susceptibility of roots to pathogen infection and disease development are unknown. Research, therefore, was carried out to evaluate the disease development in sugar beet roots caused by two common storage pathogens as a function of storage duration and storage temperature, and to identify changes in the expression of defense genes that may be influencing the root susceptibility to disease. To evaluate root susceptibility to disease, freshly harvested roots were inoculated with Botrytis cinerea or Penicillium vulpinum on the day of harvest or after 12, 40, or 120 d storage at 5 or 12 °C and the weight of rotted tissue present in the roots after incubation for 35 d after inoculation were determined. Disease susceptibility and progression to B. cinerea and P. vulpinum increased with storage duration with elevations in susceptibility occurring more rapidly to B. cinerea than P. vulpinum. Also, B. cinerea was more aggressive than P. vulpinum and caused greater rotting and tissue damage in postharvest sugar beet roots. Storage temperature had minimal effect on root susceptibility to these rot-causing pathogens. Changes in defense gene expression were determined by sequencing mRNA isolated from uninoculated roots that were similarly stored for 12, 40 or 120 d at 5 or 12 °C. As susceptibility to rot increased during storage, concurrent changes in defense-related gene expression were identified, including the differential expression of 425 pathogen receptor and 275 phytohormone signal transduction pathway-related genes. Furthermore, plant resistance and hormonal signaling genes that were significantly altered in expression coincident with the change in root susceptibility to storage rots were identified. Further investigation into the function of these genes may ultimately elucidate methods by which storage rot resistance in sugar beet roots may be improved in the future., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Key Message: Two novel and one known genomic regions associated with R-gene resistance to Phytophthora sojae were identified by genome-wide association analysis and linkage analysis in soybean. Phytophthora root and stem rot (PRR) caused by Phytophthora sojae is a severe disease that causes substantial economic losses in soybean [Glycine max (L.) Merr.]. The primary approach for successful disease management of PRR is using R-gene-mediated resistance. Based on the phenotypic evaluation of 376 cultivated soybean accessions for the R-gene type resistance to P. sojae (isolate 2457), a genome-wide association analysis identified two regions on chromosomes 3 and 8. The most significant genomic region (20.7-21.3 Mbp) on chromosome 8 was a novel resistance locus where no Rps gene was previously reported. Instead, multiple copies of the UDP-glycosyltransferase superfamily protein-coding gene, associated with disease resistance, were annotated in this new locus. Another genomic region on chromosome 3 was a well-known Rps cluster. Using the Daepung × Ilpumgeomjeong RIL population, a linkage analysis confirmed these two resistance loci and identified a resistance locus on chromosome 2. A unique feature of the resistance in Ilpumgeomjeong was discovered when phenotypic distribution was projected upon eight groups of RILs carrying different combinations of resistance alleles for the three loci. Interestingly, the seven groups carrying at least one resistance allele statistically differed from the other with none, regardless of the number of resistance alleles. This suggests that the respective three different resistance genes can confer resistance to P. sojae isolate 2457. Deployment of the three regions via marker-assisted selection will facilitate effectively improving resistance to particular P. sojae isolates in soybean breeding programs., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The common rust disease of maize is caused by the obligate biotrophic fungus Puccinia sorghi. The maize Rp1-D allele imparts resistance against the P. sorghi IN2 isolate by initiating a defense response that includes a rapid localized programmed cell death process, the hypersensitive response (HR). In this study, to identify AvrRp1-D from P. sorghi IN2, we employed the isolation of haustoria, facilitated by a biotin-streptavidin interaction, as a powerful approach. This method proves particularly advantageous in cases where the genome information for the fungal pathogen is unavailable, enhancing our ability to explore and understand the molecular interactions between maize and P. sorghi. The haustorial transcriptome generated through this technique, in combination with bioinformatic analyses such as SignalP and TMHMM, enabled the identification of 251 candidate effectors. We ultimately identified two closely related genes, AvrRp1-D.1 and AvrRp1-D.2, which triggered an Rp1-D-dependent defense response in Nicotiana benthamiana. AvrRp1-D-induced Rp1-D-dependent HR was further confirmed in maize protoplasts. We demonstrated that AvrRp1-D.1 interacts directly and specifically with the leucine-rich repeat (LRR) domain of Rp1-D through yeast two-hybrid assay. We also provide evidence that, in the absence of Rp1-D, AvrRp1-D.1 plays a role in suppressing the plant immune response. Our research provides valuable insights into the molecular interactions driving resistance against common rust in maize., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Given the negative fitness effects that pathogens impose on their hosts, the benefits of resistance should be universal. However, there is marked variation across plant species in the number of nucleotide-binding leucine-rich repeat receptors, which form a cornerstone of defense. The growth-defense trade-off hypothesis predicts costs associated with defense investment to generate variation in these traits. Our analysis comparing features of the intracellular immune-receptor repertoires with trait data of 187 species shows that in wild plants, the size of the molecular defense repertoire correlates negatively with growth. By contrast, we do not find evidence for a growth-defense trade-off in agricultural plants. Our cross-species approach highlights the central role of defense investment in shaping ecological trait variation and its sensitivity to domestication.

The Tomato Brown Rugose Fruit Virus (ToBRFV) is a pathogen that mostly affects plants from the Solanaceae family. This virus severely affects the yield of tomato (Solanum lycopersicum L.) plants, thus creating an urgent need to research the basis of resistance to manage the disease. To understand the molecular basis of resistance, we employed omics-based approaches involving leaf ionomics and transcriptomics to help us decipher the interaction between elemental and nutritional composition and investigate its effect on the gene expression profile upon the ToBRFV infection in tomatoes. Ionomics was used to investigate the accumulation of trace elements in leaves, showcasing that the plants resistant to the virus had significantly higher concentrations of iron (p-value = 0.039) and nickel (p-value = 0.042) than the susceptible ones. By correlating these findings with transcriptomics, we identified some key genes involved in iron homeostasis and abscisic acid pathways, potentially responsible for conferring resistance against the pathogen. From the obtained list of differentially expressed genes, a panel of mutation profile was discovered with three important genes-Solyc02g068590.3.1 (K+ transporter), Solyc01g111890.3.1 (LRR), and Solyc02g061770.4.1 (Chitinase) showing persistent missense mutations. We ascertain the role of these genes and establish their association with plant resistance using genotyping assays in various tomato lines. The targeted selection of these genetic determinants can further enhance plant breeding and crop yield management strategies., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Thakare et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The interactions among viruses and host plants are complex and fascinating because these organisms interact with and adapt to each other continuously. Many plant transcription factors play important roles in plant growth and development and in the resistance to viral infection. To facilitate the infection of plants, some viral proteins typically target and inhibit the function of plant transcription factors. In this study, we found an interesting phenomenon wherein the p3a protein of barley yellow dwarf virus (BYDV) can interact with the zinc finger domain of the TaDOF transcription factor in wheat; the zinc finger domain of TaDOF can interact with the promoter of TaHSP70 and inhibit the transcription of the TaHSP70 gene; and p3a interacts with the TaDOF zinc finger domain through competitive binding, alleviating TaDOF zinc finger domain-mediated inhibition of the TaHSP70 promoter, thereby promoting TaHSP70 expression and promoting infection by BYDV. This study demonstrates that BYDV p3a is an immunosuppressive factor and enriches our understanding of the pathogenesis of BYDV., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Tian et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

OsGADD45a1, a member of the growth arrest and DNA damage-inducible 45 (GADD45) family in rice, has a newly identified homologue, OsGADD45a2, which differs from OsGADD45a1 in only three amino acids. The role and function of the OsGADD45a2 in DNA demethylation are not well-understood and were investigated in this study. Osgadd45a2 mutants exhibited reduced height, shorter panicle length, fewer grains per panicle, and a lower seed setting rate compared with wild-type plants. Moreover, the results showed that OsGADD45a2 negatively regulates rice blast fungus resistance and exhibited high expression in various tissues. Using the 3000 Rice Genomes Project database, we identified four major haplotypes (each with over 100 cultivars) based on single-nucleotide polymorphisms in the coding sequence of OsGADD45a2 . Among these, Hap4 was associated with a significantly greater plant height than Hap1-3, possibly due to a functional alteration of OsGADD45a2 linked to the SNP at position 2614993. In OsGADD45a2 overexpression lines, significant decreases in CG and CHG methylation levels were observed in protein-coding genes, leading to their upregulation. Overall, our findings indicate that OsGADD45a2 acts as a methylation regulator, mediating the expression of genes essential for plant growth and development and blast resistance.

Background: Stem rot, caused by the soil-borne pathogen Sclerotium rolfsii, pose a serious challenge in the groundnut (Arachis hypogaea L) cultivation. Although this disease is widespread globally but had most adverse impact in groundnut growing regions of United States, India, and Australia. The pathogen primarily targets the crown region of the plant, resulting in systemic collapse and potentially leading to yield losses up to 80%. Effective genetic control measures are essential to mitigate the impact of this disease on groundnut production. Realizing the time and resource-consuming complex field-based phenotyping, the availability of easy and repeatable phenotyping methods may fasten the process of donor and gene discovery efforts., Results: Multi-season phenotyping was performed for stem rot on 184 minicore germplasm accessions, including checks, under two conditions: sick field screening and response to oxalic acid assay. This study demonstrated medium to high heritability (52-63% broad-sense heritability) and significant environmental influence (36%). The response to the oxalic acid assay showed a high proportion of similarity (approximately 80%) with the percent mortality observed in the sick field indicating an easy way of performing precise phenotyping. Notably, seven genotypes-ICG163, ICG721, ICG10479, ICG875, ICG11457, ICG111, and ICG2857-exhibited stable resistance, with less than 30% mortality against stem rot disease. Among these, ICG163, ICG875, and ICG111 displayed low mortality and consistent stability across multiple seasons in both the sick field and controlled conditions of the oxalic acid assay., Conclusions: The oxalic acid assay developed in this study effectively complements field phenotyping, as a reliable method for assessing stem rot resistance. Seven resistant genotypes identified through this assay can be utilized for the introgression of stem rot resistance into elite genotypes. Given the significant influence of the environment on stem rot resistance, it is essential to implement multi-season phenotyping to obtain precise results. Furthermore, the response to oxalic acid serves as a valuable supplement to traditional field phenotyping, since maintaining uniform disease pressure during field screenings is often challenging., (© 2024. The Author(s).)

Type 2C protein phosphatases (PP2Cs) constitute a large family in most plant species, but relatively few of them have been implicated in immunity. To identify and characterize PP2C phosphatases that affect tomato (Solanum lycopersicum) immunity, we generated loss-of-function mutations in 11 PP2C-encoding genes whose expression is altered in response to immune elicitors or pathogens. We report that 2 closely related PP2C phosphatases, PP2C immunity-associated candidate 3 (Pic3) and Pic12, are involved in regulating resistance to the bacterial pathogen Pseudomonas syringae pv. tomato (Pst). Loss-of-function mutations in Pic3 led to enhanced resistance to Pst in older but not younger leaves, whereas such mutations in Pic12 resulted in enhanced resistance in both older and younger leaves. Overexpression of Pic3 and Pic12 proteins in leaves of Nicotiana benthamiana inhibited resistance to Pst, and this effect was dependent on Pic3/12 phosphatase activity and an N-terminal palmitoylation motif associated with localization to the cell periphery. Pic3, but not Pic12, had a slight negative effect on flagellin-associated reactive oxygen species generation, although their involvement in the response to Pst appeared independent of flagellin. RNA-sequencing analysis of Rio Grande (RG)-PtoR wild-type plants and 2 independent RG-pic3 mutants revealed that the enhanced disease resistance in RG-pic3 older leaves is associated with increased transcript abundance of multiple defense-related genes. RG-pic3/RG-pic12 double-mutant plants exhibited stronger disease resistance than RG-pic3 or RG-pic12 single mutants. Together, our results reveal that Pic3 and Pic12 negatively regulate tomato immunity in an additive manner through flagellin-independent pathways., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Leaf rust, caused by Puccinia triticina Erikss. (Pt), is a serious disease threatening wheat (Triticum aestivum L.) production worldwide. Hydrogen peroxide (H2O2) triggered by Pt infection in resistant wheat cultivars cause oxidative damage directly to biomolecules or is activated by calcium signaling and mediates the hypersensitive response. Calmodulin-binding transcriptional activator 4 (TaCAMTA4) has been reported to negatively regulate wheat resistance to Pt. In this study, we found that TaCAMTA4 was induced by Pt race 165 in its compatible host harboring the Pt-resistant locus Lr26, TcLr26, and silencing of TaCAMTA4 increased local H2O2 accumulation and Pt resistance. Subcellular localization and autoactivation tests revealed that TaCAMTA4 is a nucleus-localized transcriptional activator. Furthermore, 4 DNA motifs recognized by TaCAMTA4 were identified by transcription factor-centered Y1H. Through analyzing the transcriptome database, 4 gene clusters were identified, each containing a different DNA motif on each promoter. Among them, the expression of catalase 1 (TaCAT1) with motif-1 was highly induced in the compatible interaction and was decreased when TaCAMTA4 was silenced. The results of electrophoretic mobility shift assay, ChIP-qPCR, and RT-qPCR further showed that TaCAMTA4 directly bound motif-1 in the TaCAT1 promoter. Furthermore, silencing of TaCAT1 resulted in enhanced resistance to Pt and increased local H2O2 accumulation in wheat, which is consistent with that of TaCAMTA4. Since calmodulin-binding transcription activators are Ca2+ sensors and catalases catalyze the decomposition of H2O2, we hypothesize that Ca2+ regulates the plant immune networks that are controlled by H2O2 and implicate a potential mechanism for Pt to suppress resistance by inducing the expression of the TaCAMTA4-TaCAT1 module, which consequently enhances H2O2 scavenging and attenuates H2O2-dependent resistance., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Jujube witches' broom (JWB) is a phytoplasma disease that causes severe damage to jujube (Ziziphus jujuba) crops worldwide. Diseased jujube plants show enhanced vegetative growth after floral reversion, including leafy flower structures (phyllody) and the fourth whorl converting into a vegetative shoot. In previous research, secreted JWB protein 3 (SJP3) was identified as an inducer of phyllody. However, the molecular mechanisms of SJP3-mediated pistil reversion remain unknown. Here, the effector SJP3 was found to interact with the MADS-box protein SHORT VEGETATIVE PHASE 3 (ZjSVP3). ZjSVP3 was expressed in young leaves and during the initial flower bud differentiation of healthy jujube-bearing shoots but was constitutively expressed in JWB phytoplasma-infected flowers until the later stage of floral development. The SJP3 effector showed the same expression pattern in the diseased buds and promoted ZjSVP3 accumulation in SJP3 transgenic jujube calli. The N-terminal domains of ZjSVP3 contributed to its escape from protein degradation in the presence of SJP3. Heterologous expression of ZjSVP3 in Nicotiana benthamiana produced typical pistil abnormalities, including trichome-enriched style and stemlike structures within the leaflike ovary, which were consistent with those in the mildly malformed lines overexpressing SJP3. Furthermore, ectopic expression of ZjSVP3 directly bound to the zinc finger protein 8 (ZjZFP8) and MADS-box gene SHATTERPROOF 1 (ZjSHP1) promoters to regulate their expression, resulting in abnormal pistil development. Overall, effector SJP3-mediated derepression of ZjSVP3 sustained its expression to interfere with pistil development, providing insight into the mechanisms of pistil reversion caused by JWB phytoplasma in specific perennial woody plant species., Competing Interests: Conflict of interest statement. The authors declare they have no conflicts of interest in this paper., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Bitter gourd is increasingly being recognized for its value as a vegetable and medicinal use, but the molecular mechanisms of pathogen resistance remain relatively poorly understood. The serine carboxypeptidase-like (SCPL) protein family plays a key role in plant growth, pathogen defense, and so on. However, a comprehensive identification and functional characterization of the SCPL gene family has yet to be conducted in bitter melon. In this study, 32 SCPL genes were identified in bitter gourd and divided into three classes. The number of SCPL genes contained in the three clusters was 7, 7, and 18, respectively. Most SCPL gene promoters contain cis -acting elements with light, hormone, and stress responses. The RNA sequencing data showed that the expression of several SCPL genes changed significantly after pathogen infection. In particular, expression of the McSCPL4 , 10 , 17 , 22 , and 25 genes increased substantially in the resistant varieties after infection, and their expression levels were higher than those in the susceptible varieties. These results suggested that genes such as McSCPL4 , 10 , 17 , 22 , and 25 may play a significant role in conferring resistance to fungal infections. Moreover, the expression levels of the McSCPL10 , 17 , 22 , 23, and 25 genes were likewise significantly changed after being induced by salicylic acid (SA) and jasmonic acid (JA). In situ hybridization showed that McSCPL22 was expressed in the vascular tissues of infected plants, which largely overlapped with the location of Fusarium oxysporum f. sp. Momordicae (FOM) infection and the site of hydrogen peroxide production. Our results showed that McSCPL22 may be involved in the regulation of the SA and JA pathways and enhance resistance to FOM in bitter gourd plants. This is the first study to perform SCPL gene family analysis in bitter gourd. McSCPL22 may have the potential to enhance FOM resistance in bitter gourd, and further investigation into its function is warranted. The results of this study may enhance the yield and molecular breeding of bitter gourd.

Ustilago crameri is a pathogenic basidiomycete fungus that causes foxtail millet kernel smut (FMKS), a devastating grain disease in most foxtail millet growing regions of the world. Carbohydrate-Binding Modules (CBMs) are one of the important families of carbohydrate-active enzymes (CAZymes) in fungi and play a crucial role in fungal growth and development, as well as in pathogen infection. However, there is little information about the CBM family in U. crameri . Here, 11 CBM members were identified based on complete sequence analysis and functional annotation of the genome of U. crameri . According to phylogenetic analysis, they were divided into six groups. Gene structure and sequence composition analysis showed that these 11 UcCBM genes exhibit differences in gene structure and protein motifs. Furthermore, several cis-regulatory elements involved in plant hormones were detected in the promoter regions of these UcCBM genes. Gene ontology (GO) enrichment and protein-protein interaction (PPI) analysis showed that UcCBM proteins were involved in carbohydrate metabolism, and multiple partner protein interactions with UcCBM were also detected. The expression of UcCBM genes during U. crameri infection is further clarified, and the results indicate that several UcCBM genes were induced by U. crameri infection. These results provide valuable information for elucidating the features of U. crameri CBMs' family proteins and lay a crucial foundation for further research into their roles in interactions between U. crameri and foxtail millet.

Jujube witches' broom, caused by phytoplasma, is a destructive disease of Chinese jujube. Studies have shown that zju-miR156s play an important role in phytoplasma infection in jujube, but the regulatory mechanism between zju-miR156c and witches' broom remains unexplored. In the current study, miRNA-seq and gene expression analysis showed that zju-miR156c was more highly induced in infected jujube plants than the other miRNAs and its target gene was ZjSPL3. In addition, the expression levels of thymidylate kinase gene (TMK JWB ) and secreted jujube protein (SJP1 JWB ) in diseased materials were higher than those in healthy controls. The expression level of zju-miR156c was significantly upregulated, while ZjSPL3 was sharply downregulated and the content of cytokinin (CTK) significantly increased. Overexpression of zju-miR156c in Arabidopsis significantly reduced the expression of AtSPL10 (homologous gene of ZjSPL3) but increased the content of CTK, and the transgenic plants exhibited witches' broom symptoms. In addition, yeast two-hybrid and co-immunoprecipitation assays confirmed that SJP1 JWB interacted with ZjERF18. Yeast one-hybrid analysis showed that ZjERF18 could interact with the promoter of zju-MIR156c. In conclusion, our results demonstrated a novel pathogenic module of ZjERF18-zju-miR156c-ZjSPL3-CTK has an important function in the formation of witches' broom caused by SJP1 JWB ., (© 2024 The Author(s). Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.)

Northern corn leaf blight, caused by Exserohilum turcicum (E. turcicum), is one of the most destructive diseases in maize, leading to serious yield losses. However, the underlying molecular mechanisms of E. turcicum infection response in maize remain unclear. In this study, we performed comparative transcriptome analysis in resistant maize inbred line J9D207 (R) and susceptible maize inbred line PH4CV (S) after infecting with E. turcicum at 0 h, 24 h and 72 h, respectively. Compared with 0 h, 9656 (24 h) and 8748 (72 h) differentially expressed genes (DEGs) were identified in J9D207, and 7915 (24 h) and 7865 (72 h) DEGs were identified in PH4CV. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that alpha-linolenic acid metabolism, benzoxazinoid biosynthesis, flavonoid biosynthesis and phenylpropanoid biosynthesis might be involved in maize defense reactions. Some DEGs coded for transcription factors, such as MYB-related, ERF, NAC, bZIP, bHLH and WRKY families, which indicated that they may participate in resistance against E. turcicum. In addition, DEGs involved in SA, JA, ABA and ET signaling pathways were revealed. Moreover, 75 SOD activity-related genes and 421 POD activity-related genes were identified through weighted gene co-expression network analysis (WGCNA), respectively. These results provide a novel insight into the resistance mechanism of maize in response to E. turcicum inoculation., (© 2024 Scandinavian Plant Physiology Society.)

Background: Bacterial wilt (BW), caused by Ralstonia solanacearum (Ral), results in substantial yield losses in pepper crops. Developing resistant pepper varieties through breeding is the most effective strategy for managing BW. To achieve this, a thorough understanding of the genetic information connected with resistance traits is essential. Despite identifying three major QTLs for bacterial wilt resistance in pepper, Bw1 on chromosome 8, qRRs-10.1 on chromosome 10, and pBWR-1 on chromosome 1, the genetic information of related BW pepper varieties has not been sufficiently studied. Here, we resequenced two pepper inbred lines, C. annuum 'MC4' (resistant) and C. annuum 'Subicho' (susceptible), and analyzed genomic variations through SNPs and Indels to identify candidate genes for BW resistance., Results: An average of 139.5 Gb was generated among the two cultivars, with coverage ranging from 44.94X to 46.13X. A total of 8,815,889 SNPs was obtained between 'MC4' and 'Subicho'. Among them, 31,190 (0.35%) were non-synonymous SNPs (nsSNPs) corresponding to 10,926 genes, and these genes were assigned to 142 Gene Ontology (GO) terms across the two cultivars. We focused on three known BW QTL regions by identifying genes with sequence variants through gene set enrichment analysis and securing those belonging to high significant GO terms. Additionally, we found 310 NLR genes with nsSNP variants between 'MC4' (R) and 'Subicho' (S) within these regions. Also, we performed an Indel analysis on these genes. By integrating all this data, we identified eight candidate BW resistance genes, including two NLR genes with nsSNPs variations in qRRs-10.1 on chromosome 10., Conclusion: We identified genomic variations in the form of SNPs and Indels by re-sequencing two pepper cultivars with contrasting traits for bacterial wilt. Specifically, the four genes associated with pBWR-1 and Bw1 that exhibit both nsSNP and Indel variations simultaneously in 'Subicho', along with the two NLR genes linked to qRRs-10.1, which are known for their direct involvement in immune responses, are identified as most likely BW resistance genes. These variants in leading candidate genes associated with BW resistance can be used as important markers for breeding pepper varieties., (© 2024. The Author(s).)

Blue light enhances the susceptibility of Nicotiana benthamiana to Phytophthora infestans, a causative agent of late blight disease. Investigating how blue light affects potato late blight resistance is an interesting aspect of exploring new ways to control late blight disease. Blue light photoreceptor phototropins (phot1, phot2) and their downstream interact protein StNRL1 have been shown to negatively regulate late blight resistance. In order to investigate whether other potato NPH3/RPT2-Like (NRL) family members are involved in regulating late blight resistance, this study focused on the potato NRL proteins containing RxSxS motif at the C-terminus. Another potato NRL protein StNRL-9, containing RxSxS motifs, was found to negatively regulate P. infestans resistance in potato and N. benthamiana. Overexpression of StNRL-9 in potato and N. benthamiana suppresses the accumulation of reactive oxygen species (ROS) and expression of the PTI marker genes NbWRKY7 and NbWRKY8. Similar to StNRL1, StNRL-9 interacts with the blue light receptors Stphot1 and Stphot2 on the cell membrane and could promote the degradation of a positive immune regulator StSWAP70. However StNRL-9 does not inhibit INF1-mediated cell death (ICD), which is different from the StNRL1 that inhibits ICD, indicating that both StNRL1 and StNRL-9 inhibit plant immunity in diverse ways. This study provides valuable information for further exploration of how plant phototropins and NRL family proteins regulate plant immunity., (© 2024 Scandinavian Plant Physiology Society.)

Autophagy is vital for plant defense against pathogens, with ATG6 being a key gene in this process. At present, little has been reported on the potential function and molecular mechanisms of ATG6 mediated pathogen resistance in pear. This study investigates the function of the pear homolog of ATG6 (PbrATG6) in resistance to Botryosphaeria dothidea. PbrATG6 is expressed differentially in pear tissues and its expression increases upon infection. Overexpression of PbrATG6 enhances resistance in Arabidopsis and pear calli, while silencing it increases susceptibility. PbrTLP15, a pathogenesis-related protein belonging to the PR5 family, was found that interacts with PbrATG6 by a yeast two-hybrid screening. Yeast two-hybrid, luciferase complementation imaging, bimolecular fluorescence complementation assays and pull-down assays showed that PbrATG6 interacts with PbrTLP15. The transient silencing transgenic assays of PbrATG6 and PbrTLP15 revealed that PbrATG6 could cooperate with PbrTLP15 to regulate pear B. dothidea resistance. In addition, transcriptional analyses of autophagy key genes in pTRV-PbrTLP15 and transmission electron microscopy (TEM) assays also implied that PbrTLP15 does affect autophagy. Hence, PbrATG6 and PbrTLP15 may synergistically enhance pear B. dothidea disease resistance. It provides a new strategy for the study of autophagy in pear disease resistance and enriches the research on pear disease resistance mechanism., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Shade is a stressful factor for most plants, leading to both morphological and physiological changes, and often resulting in increased susceptibility to diseases and pathogen attacks. Our study revealed that the isoflavonoid synthesis pathway was inhibited in alfalfa under shade, resulting in a significant reduction in disease resistance. Overexpression of MsIFS1, a switch regulator in isoflavonoid synthesis, led to a notable increase in endogenous isoflavonoids and enhanced resistance to Pseudomonas syringae pv. tomato DC3000 (Pst. DC3000). Conversely, MsIFS1-RNAi had the opposite effect. Yeast one-hybrid (Y1H) assays revealed that the shade-responsive transcription factor MsWRKY41 could directly bind to the MsIFS1 promoter. This interaction was confirmed through Dual-Luciferase Reporter (Dual-LUC) and Chromatin Immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) assays, both in vitro and in vivo. Overexpression of MsWRKY41 not only enhanced alfalfa's resistance to Pst. DC3000 but also promoted the accumulation of isoflavonoids. Additionally, yeast two-hybrid (Y2H) assays showed that neither MsWRKY41 nor MsIFS1 physically interacted with the Type III effector (T3SE) HopZ1 secreted by Pst. DC3000, suggesting that the MsWRKY41-MsIFS1 module is not a direct target of HopZ1. These findings provide valuable theoretical insights and genetic resources for the development of shade-tolerant alfalfa with enhanced disease resistance., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

The cuticle is the first physical barrier covering the surface of tomatoes and plays an important role in multiple stress responses. But the molecular regulatory networks of cuticle formation are not fully understood. In this study, we found that SlMYB72 can interact with SlTAGL1 to regulate the formation of fruit cuticle in tomato. Downregulating the expression of SlMYB72 inhibits the formation of fruit cuticle, resulting in a reduced fruit cuticle thickness, accelerated postharvest water loss, and increased susceptibility to Botrytis cinerea. RNA sequencing analysis showed that downregulation of the SlMYB72 gene decreased the expression levels of genes related to fatty acid and cuticle metabolism. SlMYB72 regulates the cuticle formation by directly binding to the promoter of long-chain acyl-coA synthetases (SlLACS1) and medium-chain alkane hydroxylase (SlMAH1). Moreover, SlMYB72 interacts with SlTAGL1, which can enhance the transcriptional activation of SlMYB72 on the SlMAH1 promoter. Overall, our study expands our understanding of the regulation of cuticle formation by SlMYB72 and provides new insights into fruit shelf life extension via manipulation of cuticle content., (© 2024 Society for Experimental Biology and John Wiley & Sons Ltd.)

Carrot (Daucus carota) is one of the most popular and nutritious vegetable crops worldwide. However, significant yield losses occur every year due to leaf blight, a disease caused by a fungal pathogen (Alternaria dauci). Past research on resistance to leaf blight disease in carrots has been slow because of the low-quality genome assemblies of both carrot and the pathogen. Here, we report the greatly improved assemblies and annotations of telomere-to-telomere (T2T) reference genomes of carrot DH13M14 (451.04 Mb) and A. dauci A2016 (34.91 Mb). Compared with the previous carrot genome versions, our assembly featured notable improvements in genome size, continuity, and completeness of centromeres and telomeres. In addition, we generated a time course transcriptomic atlas during the infection of carrots by A. dauci and captured their dynamic gene expression reprogramming during the interaction process. During infection, A. dauci genes encoding effectors and enzymes responsible for the degradation of plant cell wall components, e.g., cellulose and pectin, were identified, which appeared to increase pathogenic ability through upregulation. In carrot, the coordinated gene expression of components of pattern- and effector-triggered immunity (PTI and ETI) in response to A. dauci attack was characterized. The biosynthesis or signal transduction of plant hormones, including JA, SA, and ethylene, was also involved in the carrot response to A. dauci. This work provides a foundation for understanding A. dauci pathogenic progression and carrot defense mechanisms to improve carrot resistance to leaf blight disease. The Carrot Database (CDB) developed also provides a useful resource for the carrot community., (© 2024 The Author(s). The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

Plant trichomes are an excellent model for studying cell differentiation and development, providing crucial defenses against biotic and abiotic stresses. There is a well-established inverse relationship between trichome density and aphid prevalence, indicating that higher trichome density leads to reduced aphid infestations. Here we present the cloning and characterization of a dominant quantitative trait locus, HIC (hirsute cotton), which significantly enhances cotton trichome density. This enhancement leads to markedly improved resistance against cotton aphids. The HIC encodes an HD-ZIP IV transcriptional activator, crucial for trichome initiation. Overexpression of HIC leads to a substantial increase in trichome density, while knockdown of HIC results in a marked decrease in density, confirming its role in trichome regulation. We identified a variant in the HIC promoter (-810 bp A to C) that increases transcription of HIC and trichome density in hirsute cotton compared with Gossypium hirsutum cultivars with fewer or no trichomes. Interestingly, although the -810 variant in the HIC promoter is the same in G. barbadense and hirsute cotton, the presence of a copia-like retrotransposon insertion in the coding region of HIC in G. barbadense causes premature transcription termination. Further analysis revealed that HIC positively regulates trichome density by directly targeting the EXPANSIN A2 gene, which is involved in cell wall development. Taken together, our results underscore the pivotal function of HIC as a primary regulator during the initial phases of trichome formation, and its prospective utility in enhancing aphid resistance in superior cotton cultivars via selective breeding., (© 2024 Society for Experimental Biology and John Wiley & Sons Ltd.)

Canker disease caused by the bacterium Lonsdalea populi is one of the most destructive diseases affecting poplar stems. However, the detailed stress response mechanisms of poplar have not been widely characterized. To explore the diverse regulatory RNA landscape and the function of key regulators in poplar subjected to L. populi stress, we integrated time-course experiment with mock-inoculation (CK) and inoculation (IN) with L. populi at the first, third, and sixth day (IN1, IN3, IN6) on Populus × euramericana cv. '74/76' (107), small RNA-seq, whole transcriptome-wide analysis, degradome analysis and transgenic experiments. A total of 98 differentially expressed (DE) miRNA, 17 974 DEmRNA, and 807 DElncRNA were identified in poplar infected by L. populi, presenting dynamic changes over the infection course. Regulatory networks among RNAs were further constructed. Notably, a network centered on ptc-miR482a in CK-vs-IN3 contained most DEGs. We show that miR482a and miR1448 are located in one transcript as a polycistron. Overexpression of pre-miR482a-miR1448 (OX482-1448) and pre-miR482a (OX482) increased poplar susceptibility to canker pathogen with reduced accumulation of reactive oxygen species, while the suppression of miR482a (STTM482) conferred poplar disease resistance. PHA7 was validated as the target of miR482a with degradome sequencing and tobacco transient co-transformation, its expression being downregulated in OX482-1448 and OX482 lines. Additionally, a series of phasiRNAs were triggered by miR482a targeting PHA7, forming regulatory cascades with more RLP, NBS-LRR, and PK genes, further verifying the defense function of miR482a. These findings provide insights for understanding the roles of ncRNAs and regulatory networks involved in poplar immunity., (© 2024 Society for Experimental Biology and John Wiley & Sons Ltd.)

The Ribosomal protein L18 (RPL18) protein gene family plays an important role in plant growth, development and stress response. Although the RPL18 genes have been identified in several plant species, the RPL18 gene family in wheat (Triticum aestivum) is still unexplored. This study found 8 TaRPL18 genes, each of which has a significantly different gene sequence length and is evenly distributed on the chromosome; Additionally, these proteins have similar physicochemical characteristics as well as secondary and tertiary structures. 17 RPL18 genes in 4 species (wheat, Arabidopsis, rice, and maize) were classified into 5 groups, and the TaRPL18 genes within the same group showed similar structures and conserved motifs. Analysis of the cis-acting elements in the TaRPL18 genes promoter regions revealed the presence of developmental and stress-responsive elements in the majority of the genes. Through yeast two-hybrid (Y2H) experiments, it was confirmed that the powdery mildew resistance protein TaPm46 physically interacts with the Class IV TaRPL18-1. Functional analysis indicated that TaRPL18-1-silenced wheat plants show reduced resistance to powdery mildew compared to the wild type (WT), with decreased expression levels of PAL and PPO genes, and increased expression levels of the PR gene. The findings of this study provide a basis for clarifying the function of the TaRPL18 genes and will be useful for the selection of disease-resistant varieties of wheat., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In plants, RNA silencing constitutes a strong defense against viral infection, which viruses counteract with RNA-silencing suppressors (RSSs). Understanding the interactions between viral RSSs and host factors is crucial for elucidating the molecular arms race between viruses and host plants. We report that the helicase motif (Hel) of the replicase encoded by apple stem grooving virus (ASGV)-the main virus affecting pear trees in China-is an RSS that can inhibit both local and systemic RNA silencing, possibly by binding double-stranded (ds) siRNA. The transcription factor related to ABSCISIC ACID INSENSITIVE3/VIVIPAROUS1 from pear (PbRAV1) enters the cytoplasm and binds Hel through its C terminus, thereby attenuating its RSS activity by reducing its binding affinity to 21- and 24-nt ds siRNA, and suppressing ASGV infection. PbRAV1 can also target p24, an RSS encoded by grapevine leafroll-associated virus 2 (GLRaV-2), with similar negative effects on p24's suppressive function and inhibition of GLRaV-2 infection. Moreover, like the positive role of the PbRAV1 homolog from grapevine (VvRAV1) in p24's previously reported RSS activity, ASGV Hel can also hijack VvRAV1 and employ the protein to sequester 21-nt ds siRNA, thereby enhancing its own RSS activity and promoting ASGV infection. Furthermore, PbRAV1 neither interacts with CP, an RSS encoded by grapevine inner necrosis virus, nor has any obvious effect on CP's RSS activity. Our results identify an RSS encoded by ASGV and demonstrate that PbRAV1, representing a novel type of RAV transcription factor, plays a defensive role against viral infection by targeting viral RSSs., (© 2024 Society for Experimental Biology and John Wiley & Sons Ltd.)

Plants use nucleotide-binding leucine-rich repeat receptors (NLRs) to sense pathogen effectors, initiating effector-triggered immunity (ETI). NLRs containing RESISTANCE TO POWDERY MILDEW 8 domain (RNLs) function as "helper" NLRs in flowering plants and support the immune responses mediated by "sensor" NLRs in cooperation with lipase-EP domain fused proteins (EP proteins). Despite their crucial roles in ETI, much remains unclear about the evolutionary trajectories of RNLs and their functional partners EP proteins. Here, we perform phylogenomic analyses of RNLs in 90 plants, covering the major diversity of plants, and identify the presence of RNLs in land plants and green algae, expanding the distribution of RNLs. We uncover a neglected major RNL group in gymnosperms, besides the canonical major group with NRG1s and ADR1s, and observe a drastic increase in RNL repertoire size in conifers. Phylogenetic analyses indicate that RNLs originated multiple times through domain shuffling, and the evolution of RNLs underwent a birth-and-death process. Moreover, we trace the origin of EP proteins back to the last common ancestor of vascular plants. We find that both RNLs and EP proteins evolve mainly under negative selection, revealing strong constraints on their function. Concerted losses and positive correlation in copy number are observed between RNL and EP sublineages, suggesting their cooperation in function. Together, our findings provide insights into the origin and evolution of plant helper NLRs, with implications for predicting novel innate immune signaling modules., (© 2024 Society for Experimental Biology and John Wiley & Sons Ltd.)

Wheat (Triticum aestivum L.) is a globally staple crop vulnerable to various fungal diseases, significantly impacting its yield. Plant cell surface receptors play a crucial role in recognizing pathogen-associated molecular patterns (PAMPs) and activating PAMP-triggered immunity, boosting resistance against a wide range of plant diseases. Although the role of plant chitin receptor CERK1 in immune recognition and defense has been established in Arabidopsis and rice, its function and potential agricultural applications in enhancing resistance to crop diseases remain largely unexplored. Here, we identify and characterize TaCERK1 in Triticeae crop wheat, uncovering its involvement in chitin recognition, immune regulation, and resistance to fungal diseases. By a comparative analysis of CERK1 homologs in Arabidopsis and monocot crops, we demonstrate that AtCERK1 in Arabidopsis elicits the most robust immune response. Moreover, we show that overexpressing TaCERK1 and AtCERK1 in wheat confers resistance to multiple fungal diseases, including Fusarium head blight, stripe rust, and powdery mildew. Notably, transgenic wheat lines with moderately expressed AtCERK1 display superior disease resistance and heightened immune responses without adversely affecting growth and yield, compared to TaCERK1 overexpression transgenics. Our findings highlight the significance of plant chitin receptors across diverse plant species and suggest potential strategies for bolstering crop resistance against broad-spectrum diseases in agricultural production through the utilization of plant immune receptors., (© 2024 Society for Experimental Biology and John Wiley & Sons Ltd.)

Endogenous banana streak virus (eBSV) integrants derived from three distinct species, present in Musa balbisiana (B) but not Musa acuminata (A) banana genomes are able to reconstitute functional episomal viruses causing banana streak disease in interspecific triploid AAB banana hybrids but not in the diploid (BB) parent line, which harbours identical eBSV loci. Here, we investigated the regulation of these eBSV. In-depth characterization of siRNAs, transcripts and methylation derived from eBSV using Illumina and bisulfite sequencing were carried out on eBSV-free Musa acuminata AAA plants and BB or AAB banana plants with eBSV. eBSV loci produce low-abundance transcripts covering most of the viral sequence and generate predominantly 24-nt siRNAs. siRNA accumulation is restricted to duplicated and inverted viral sequences present in eBSV. Both siRNA-accumulating and nonaccumulating sequences of eBSV in BB plants are heavily methylated in all three CG, CHG and CHH contexts. Our data suggest that eBSVs are controlled at the epigenetic level in BB diploids. This regulation not only prevents their awakening and systemic infection of the plant but is also probably involved in the inherent resistance of the BB plants to mealybug-transmitted viral infection. These findings are thus of relevance to other plant resources hosting integrated viruses., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)