Resistance In Cucumber Research Articles

A genome-wide association study reveals molecular mechanism underlying powdery mildew resistance in cucumber

BackgroundPowdery mildew is a disease with one of the most substantial impacts on cucumber production globally. The most efficient approach for controlling powdery mildew is the development of genetic resistance; however, few genes associated with inherent variations in cucumber powdery mildew resistance have been identified as of yet.ResultsIn this study, we re-sequence 299 cucumber accessions, which are divided into four geographical groups. A genome-wide association study identifies 50 sites significantly associated with natural variations in powdery mildew resistance. Linkage disequilibrium analysis further divides these 50 sites into 32 linkage disequilibrium blocks containing 41 putative genes. Virus-induced gene silencing and gene expression analysis implicate CsGy5G015960, which encodes a phosphate transporter, as the candidate gene regulating powdery mildew resistance. On the basis of the resequencing data, we generate five CsGy5G015960 haplotypes, identifying Hap.1 as the haplotype most likely associated with powdery mildew resistance. In addition, we determine that a 29-bp InDel in the 3′ untranslated region of CsGy5G015960 is responsible for mRNA stability. Overexpression of CsGy5G015960Hap.1 in the susceptible line enhances powdery mildew resistance and phosphorus accumulation. Further comparative RNA-seq analysis demonstrates that CsGy5G015960Hap.1 may regulate cucumber powdery mildew resistance by maintaining a higher H2O2 level through the depletion of multiple class III peroxidases.ConclusionsHere we identify a candidate powdery mildew-resistant gene in cucumber using GWAS. The identified gene may be a promising target for molecular breeding and genetic engineering in cucumber to enhance powdery mildew resistance.

Cy-1, a major QTL for tomato leaf curl New Delhi virus resistance, harbors a gene encoding a DFDGD-Class RNA-dependent RNA polymerase in cucumber (Cucumis sativus)

BackgroundTomato leaf curl New Delhi virus (ToLCNDV) (family Geminiviridae, genus Begomovirus) is a significant threat to cucumber (Cucumis sativus) production in many regions. Previous studies have reported the genetic mapping of loci related to ToLCNDV resistance, but no resistance genes have been identified.ResultsWe conducted map-based cloning of the ToLCNDV resistance gene in cucumber accession No.44. Agroinfiltration and graft-inoculation analyses confirmed the resistance of No.44 to ToLCNDV isolates from the Mediterranean and Asian countries. Initial mapping involving two rounds of phenotyping with two independent F2 populations generated by crossing the begomovirus-susceptible cultivar SHF and No.44 consistently detected major quantitative trait loci (QTLs) on chromosomes 1 and 2 that confer resistance to ToLCNDV. Fine-mapping of Cy-1, the dominant QTL on chromosome 1, using F3 populations narrowed the candidate region to a 209-kb genomic segment harboring 24 predicted genes. Among these genes, DFDGD-class RNA-dependent RNA polymerase (CsRDR3), an ortholog of Ty-1/Ty-3 of tomato and Pepy-2 of capsicum, was found to be a strong candidate conferring ToLCNDV resistance. The CsRDR3 sequence of No.44 contained multiple amino acid substitutions; the promoter region of CsRDR3 in No.44 had a large deletion; and the CsRDR3 transcript levels were greater in No.44 than in SHF. Virus-induced gene silencing (VIGS) of CsRDR3 using two chromosome segment substitution lines harboring chromosome 1 segments derived from No.44 compromised resistance to ToLCNDV.ConclusionsForward and reverse genetic approaches identified CsRDR3, which encodes a DFDGD-class RNA-dependent RNA polymerase, as the gene responsible for ToLCNDV resistance at the major QTL Cy-1 on chromosome 1 in cucumber. Marker-assisted breeding of ToLCNDV resistance in cucumber will be expedited by using No.44 and the DNA markers developed in this study.

Evaluation of the efficacy of some organic substances to induce resistance in cucumber against root knot nematode, Meloidogyne spp.

The study aimed to evaluate the impact of certain biofertilizers on some growth parameters, total chlorophyll content and the concentrations of enzymes phenylalanine amino-lyase (PAL) and peroxidase (POX) pre and post infestation of cucumber with root-knot nematodes (Meloidogyne spp.). The results demonstrate that supplementing substances before or after nematode infection resulted in enhanced growth parameters of cucumbers, including shoot growth, fresh and dry weight of root, total chlorophyll content and the concentrations of the enzymes PAL and POX, with significant differences compared to non-infected plants. Among those that showed a positive effect were ascorbic acid, K2HPO4, liquid earthworms, crushed fertilizer and compost. Among the test biofertilizers evaluated, the liquid vermicompost proved to be the standout performer compared to the other treatments, demonstrating the most substantial growth improvements such as the highest increase in both shoot fresh weight (44.67 g per plant) and dry weight (5.93 g per plant). Additionally, this treatment resulted in a remarkable 83.00 % increase in total chlorophyll content and a significant increase in PAL and POX enzyme activity (18.25 and 25.29 IU g-1 respectively).

Genome-wide identification of Brassinosteroid insensitive 1-associated receptor kinase 1 genes and expression analysis in response to pathogen infection in cucumber (Cucumis sativus L.)

BackgroundBAK1 (Brassinosteroid insensitive 1-associated receptor kinase 1) plays an important role in disease resistance in plants. However, the function of BAK1 family in cucumber and the decisive genes for disease-resistance remain elusive.ResultsHere, we identified 27 CsBAK1s in cucumber, and classified them into five subgroups based on phylogenetic analysis and gene structure. CsBAK1s in the same subgroup shared the similar motifs, but different gene structures. Cis-elements analysis revealed that CsBAK1s might respond to various stress and growth regulation. Three segmentally duplicated pairwise genes were identified in cucumber. In addition, Ka/Ks analysis indicated that CsBAK1s were under positive selection during evolution. Tissue expression profile showed that most CsBAK1s in Subgroup II and IV showed constitutive expression, members in other subgroups showed tissue-specific expression. To further explore whether CsBAK1s were involved in the resistance to pathogens, the expression patterns of CsBAK1s to five pathogens (gummy stem blight, powdery mildew, downy mildew, grey mildew, and fusarium wilt) reveled that different CsBAK1s had specific roles in different pathogen infections. The expression of CsBAK1-14 was induced/repressed significantly by five pathogens, CsBAK1-14 might play an important role in disease resistance in cucumber.Conclusions27 BAK1 genes were identified in cucumber from a full perspective, which have important functions in pathogen infection. Our study provided a theoretical basis to further clarify the function of BAK1s to disease resistance in cucumber.

The synthesis of triacylglycerol by diacylglycerol acyltransferases (CsDGAT1A and CsDGAT2D) is essential for tolerance of cucumber's resistance to low-temperature stress.

CsDGAT1A and CsDGAT2D play a positive regulatory role in cucumber's response to low-temperature stress and positively regulate the synthesis of triacylglycerol (TAG). Triacylglycerol (TAG), a highly abundant and significant organic compound in plants, plays crucial roles in plant growth, development, and stress responses. The final acetylation step of TAG synthesis is catalyzed by diacylglycerol acyltransferases (DGATs). However, the involvement of DGATs in cucumber's low-temperature stress response remains unexplored. This study focused on two DGAT genes, CsDGAT1A and CsDGAT2D, investigating their function in enhancing cucumber's low-temperature stress tolerance. Our results revealed that both proteins were the members of the diacylglycerol acyltransferase family and were predominantly localized in the endoplasmic reticulum. Functional analysis demonstrated that transient silencing of CsDGAT1A and CsDGAT2D significantly compromised cucumber's low-temperature stress tolerance, whereas transient overexpression enhanced it. Furthermore, the TAG content quantification indicated that CsDGAT1A and CsDGAT2D promoted TAG accumulation. In conclusion, this study elucidates the lipid metabolism mechanism in cucumber's low-temperature stress response and offers valuable insights for the cultivation of cold-tolerant cucumber plants.

Transcription Cofactor CsMBF1c Enhances Heat Tolerance of Cucumber and Interacts with Heat-Related Proteins CsNFYA1 and CsDREB2.

Multiprotein bridging factor 1 (MBF1) is a very important transcription factor (TF) in plants, whose members influence numerous defense responses. Our study found that MBF1c in Cucurbitaceae was highly conserved. CsMBF1c expression was induced by temperature, salt stress, and abscisic acid (ABA) in cucumber. Overexpressed CsMBF1c enhanced the heat resistance of a cucumber, and the Csmbf1c mutant showed decreased resistance to high temperatures (HTs). CsMBF1c played an important role in stabilizing the photosynthetic system of cucumber under HT, and its expression was significantly associated with heat-related TFs and genes related to protein processing in the endoplasmic reticulum (ER). Protein interaction showed that CsMBF1c interacted with dehydration-responsive element binding protein 2 (CsDREB2) and nuclear factor Y A1 (CsNFYA1). Overexpression of CsNFYA1 in Arabidopsis improved the heat resistance. Transcriptional activation of CsNFYA1 was elevated by CsMBF1c. Therefore, CsMBF1c plays an important regulatory role in cucumber's resistance to high temperatures.

Mechanisms of synthetic bacterial flora YJ-1 to enhance cucumber resistance under combined phthalate-disease stresses

Biotic and abiotic stresses have emerged as major constraints to agricultural production, causing irreversible adverse impacts on agricultural production systems and thus posing a threat to food security. In this study, a new strain of Bacillus subtilis DNYB-S1 was isolated from soil contaminated with Fusarium wilt. It was found that artificially synthetic flora (YJ-1) [Enterobacter sp. DNB-S2 and Rhodococcus pyridinovorans DNHP-S2, DNYB-S1] could effectively mitigate both biotic (Fusarium wilt) and abiotic (phthalates) sources of stresses, with the inhibition rate of YJ-1 resistant to wilt being 71.25% and synergistic degradation of 500 mg/L PAEs was 91.23%. The adaptive difference of YJ-1 was 0.59 and the ecological niche overlap value was −0.05 as determined by Lotka-Volterra modeling. These results indicate that YJ-1 has good ecological stability. The major degradation intermediates included 2-ethylhexyl benzoate (EHBA), phthalic acid (PA), diisobutyl phthalate (DIBP), and butyl benzoate, suggesting that YJ-1 can provide a more efficient pathway for PAEs degradation. In addition, there was metabolic mutualism among the strains that will selectively utilize the provided carbon source (some metabolites of PAEs) for growth. The pot experiment showed that YJ-1 with cucumber reduced the incidence of cucumber wilt by 45.31%. YJ-1 could reduce the concentration of PAEs (DBP: DEHP = 1:1) in soil species from 30 mg/kg to 4.26 mg/kg within 35 d, with a degradation efficiency of 85.81%. Meanwhile, the concentration of PAEs in cucumber was reduced to 0.01 mg/kg, indicating that YJ-1 is directly involved in the degradation of soil PAEs and the enhancement of plant immunity. In conclusion, this study provides a new perspective for the development of customized microbiomes for phytoremediation under combined biotic-abiotic stresses in agricultural production processes.

Ficus carica Linn leaves extract induces cucumber resistance to Podosphaera xanthii by inhibiting conidia and regulating enzyme activity

Cucumbers (Cucumis sativus L.) are highly susceptible to infection by powdery mildew (PM), resulting in a deterioration of cucumber quality. The utilization of chemical control measures can effectively control cucumber PM; however, it may potentially give rise to issues such as environmental pollution, compromised safety, and drug resistance. The urgent need for a green, secure, and efficient strategy to manage cucumber PM necessitates attention. The predominant fungal pathogen causing PM in cucumbers is Podosphaera xanthii (P. xanthii) in this study. The preventive and control efficacy of Ficus carica Linn leaf extract (FLE) against P. xanthii was investigated for the first time. The preventive experiments entail pre-infection spraying of FLE prior to the occurrence of P. xanthii, while the curative experiments involve post-outbreak spraying of FLE. After comprehensive consideration, the optimal approach was determined to be spraying the front of cucumber leaves with a concentration of 0.04 mg/mL FLE in order to effectively control P. xanthii in leaf disc bioassay. The lowest superposition rate from FLE and trisodium phosphate dodecahydrate was 12.86 %, indicating that the impact of FLE on the respiratory metabolism of P. xanthii conidia primarily operates through the hexose monophosphate pathway (HMP). The growth of P. xanthii conidia and mycelium can be inhibited or delayed by FLE. The application of FLE can effectively mitigated chlorophyll loss, suppressed the activities of peroxidase (POD) and polyphenol oxidase (PPO), as well as enhanced the activities of phenylalanine ammonia lyase (PAL), chitinase, and β- 1,3-glucanase. Furthermore, three representative enzyme genes, PAL, POD, and chitinase, were chosen for the purpose of validation. The results suggested that FLE has the ability to modulate the expression of relevant enzyme genes, with peak effects observed after 3–4 days of treatment. The findings offer theoretical support for the advancement of novel botanical pesticides aimed at controlling crop diseases.

A multifaceted crosstalk between brassinosteroid and gibberellin regulates the resistance of cucumber to Phytophthora melonis.

Cucumber plants are highly susceptible to the hemibiotroph oomycete Phytophthora melonis. However, the mechanism of resistance to cucumber blight remains poorly understood. Here, we demonstrated that cucumber plants with impairment in the biosynthesis of brassinosteroids (BRs) or gibberellins (GAs) were more susceptible to P. melonis. By contrast, increasing levels of endogenous BRs or exogenously application of 24-epibrassinolide enhanced the resistance of cucumber plants against P. melonis. Furthermore, we found that both knockout and overexpression of the BR biosynthesis gene CYP85A1 reduced the endogenous GA3 content compared with that of wild-type plants under the condition of inoculation with P. melonis, and the enhancement of disease resistance conferred by BR was inhibited in plants with silencing of the GA biosynthetic gene GA20ox1 or KAO. Together, these findings suggest that GA homeostasis is an essential factor mediating BRs-induced disease resistance. Moreover, BZR6, a key regulator of BR signaling, was found to physically interact with GA20ox1, thereby suppressing its transcription. Silencing of BZR6 promoted endogenous GA biosynthesis and compromised GA-mediated resistance. These findings reveal multifaceted crosstalk between BR and GA in response to pathogen infection, which can provide a new approach for genetically controlling P. melonis damage in cucumber production.

Genome-wide association study identifies candidate genes for bacterial soft rot resistance in cucumber seedlings

Bacterial soft rot (BSR) caused by Pectobacterium carotovorum subsp. brasiliense (Pcb) is a serious bacterial disease which negatively impact yield and quality in cucumber. However, the genetic mechanism of BSR resistance in cucumber has not been reported. Here, we investigated the BSR resistance of 119 cucumber core germplasm worldwide at the seedling stage and identified 26 accessions highly resistant to BSR. A total of 1 642 740 single-nucleotide polymorphisms (SNPs) were used to conduct GWAS, and five loci associated with BSR resistance were detected on four chromosomes: gBSR2.1, gBSR2.2, gBSR3.1, gBSR4.1 and gBSR5.1. Based on haplotype analysis, sequence polymorphisms, functional annotation and qRT-PCR analysis, six candidate genes were identified within the five loci. CsaV3_2G014450, CsaV3_2G014490, CsaV3_2G016000, CsaV3_3G000850, CsaV3_4G033150, and CsaV3_5G000390 each had nonsynonymous SNPs, and were significantly up-regulated in the resistant genotypes after inoculation. And CsaV3_5G000390 in the susceptible genotype was significantly up-regulated after inoculation. The identification of these candidate genes lays a foundation for understanding the genetic mechanism of BSR resistance in cucumber. Generally, our study mined genes associated with BSR resistance in cucumber seedlings and will assist the breeding of BSR-resistant cucumber cultivars.

Screening of Germplasm and Construction of Evaluation System for Autotoxicity Tolerance during Seed Germination in Cucumber

Due to the widespread use of intensive cropping patterns, the problem of continuous cropping obstacle, which is dominated by autotoxicity, has been becoming more and more prominent. Although many methods have been proposed to overcome the continuous cropping obstacle of cucumber, no study has reported the screening and evaluation of cucumber germplasm resistant to autotoxicity. In this study, 28 physiological indices related to the cucumber bud stage under cinnamic acid (CA) treatment were determined. In total, 45 cucumber cultivars were classified into three groups using principal component analysis and cluster analysis, and a model for evaluating cucumber resistance to autotoxicity was developed. The evaluation model was validated using autotoxicity-tolerant and non-autotoxicity-tolerant cultivars. The results showed that the growth of non-autotoxicity-tolerant cultivars was significantly inhibited compared to autotoxicity-tolerant cultivars. This indicated that the evaluation model of cucumber autotoxicity tolerance is reliable. The results of this study provide a valuable reference for the application of cucumber autotoxicity-tolerant germplasm resources and the development of autotoxicity-tolerant genes.

Cloning and Functional Analysis of CsROP5 and CsROP10 Genes Involved in Cucumber Resistance to Corynespora cassiicola.

The cloning of resistance-related genes CsROP5/CsROP10 and the analysis of their mechanism of action provide a theoretical basis for the development of molecular breeding of disease-resistant cucumbers. The structure domains of two Rho-related guanosine triphosphatases from plant (ROP) genes were systematically analyzed using the bioinformatics method in cucumber plants, and the genes CsROP5 (Cucsa.322750) and CsROP10 (Cucsa.197080) were cloned. The functions of the two genes were analyzed using reverse-transcription quantitative PCR (RT-qPCR), virus-induced gene silencing (VIGS), transient overexpression, cucumber genetic transformation, and histochemical staining technology. The conserved elements of the CsROP5/CsROP10 proteins include five sequence motifs (G1-G5), a recognition site for serine/threonine kinases, and a hypervariable region (HVR). The knockdown of CsROP10 through VIGS affected the transcript levels of ABA-signaling-pathway-related genes (CsPYL, CsPP2Cs, CsSnRK2s, and CsABI5), ROS-signaling-pathway-related genes (CsRBOHD and CsRBOHF), and defense-related genes (CsPR2 and CsPR3), thereby improving cucumber resistance to Corynespora cassiicola. Meanwhile, inhibiting the expression of CsROP5 regulated the expression levels of ROS-signaling-pathway-related genes (CsRBOHD and CsRBOHF) and defense-related genes (CsPR2 and CsPR3), thereby enhancing the resistance of cucumber to C. cassiicola. Overall, CsROP5 and CsROP10 may participate in cucumber resistance to C. cassiicola through the ROS and ABA signaling pathways.

Fine mapping and analysis of a candidate gene controlling Phytophthora blight resistance in cucumber.

Cucumber blight is a destructive disease. The best way to control this disease is resistance breeding. This study focuses on disease resistance gene mapping and molecular marker development. We used the resistant cucumber, JSH, and susceptible cucumber, B80, as parents to construct F2 populations. Bulked segregant analysis (BSA) combined with specific length amplified fragment sequencing (SLAF-seq) were used, from which we developed cleaved amplified polymorphic sequence (CAPs) markers to map the resistance gene. Resistance in F2 individuals showed a segregation ratio of resistance:susceptibility close to 3:1. The gene in JSH resistant cucumber was mapped to an interval of 9.25 kb, and sequencing results for the three genes in the mapped region revealed three mutations at base sites 225, 302, and 591 in the coding region of Csa5G139130 between JSH and B80, but no mutations in coding regions of Csa5G139140 and Csa5G139150. The mutations caused changes in amino acids 75 and 101 of the protein encoded by Csa5G139130, suggesting that Csa5G139130 is the most likely resistance candidate gene. We developed a molecular marker, CAPs-4, as a closely linked marker for the cucumber blight resistance gene. This is the first report on mapping of a cucumber blight resistance gene and will provideg a useful marker for molecular breeding of cucumber resistance to Phytophthora blight.

You don't win friends with bad salad! A gene editing approach to enhance the powdery mildew resistance in cucumber.

You don't win friends with bad salad! A gene editing approach to enhance the powdery mildew resistance in cucumber.

Bioactive carbohydrate from potato peels: A sweet inducer that mediate resistance against powdery mildew disease of cucumber through phytohormonal alterations

Large volumes of nutritive wastes are generated by food industries, which can be used in the production of bioactive compounds of commercial values. The waste generated by potato chips industries contains high quantities of insoluble starch, high concentrations of total suspended solids and chemical oxygen demand, or environmental issues. In this study, the starch extracted from the potato peels was subjected to bioconversion of isomaltose-oligosaccharide by enzymatic method and further eluted isomatlose by liquid chromatography-mass spectrometry. The structural elucidation of isomaltose was confirmed by 1H NMR and 13C NMR spectroscopy. The purified isomaltose displayed excellent in vitro antifungal action against Erysiphe cichoracearum and Fusarium oxysporum at concentrations 1.0 and 2.5 mg/ml, respectively. Notably, cucumber-seeds primed with isomaltose (2.5 and 5.0 mg/mL) concentrations recorded a significant improvement of germination (91.5 and 89 %) and vigor (2210.93 and 2015.7) over the untreated control. Subsequently, the isomaltose-primed cucumber plants challenged with powdery mildew pathogen showed enhanced protection of 63% in comparison with pathogen alone-induced plants. This enhanced growth parameters and induced resistance in cucumber may be fairly contributed by the upregulation of phytohormone levels in primed plants observed in this study. Taken together, this study is unique in the use of bioactive isomaltose from potato peels, which could enable us to minimize the agro-industrial wastes and most importantly could help to develop new formulations for sustainable crop protection strategies.

Attenuation of Zucchini mosaic virus disease in cucumber plants by mycorrhizal symbiosis

Key messageArbuscular mycorrhizal fungi generated systemic acquired resistance in cucumber to Zucchini yellow mosaic virus, indicating their prospective application in the soil as a sustainable, environmentally friendly approach to inhibit the spread of pathogens.The wide spread of plant pathogens affects the whole world, causing several plant diseases and threatening national food security as it disrupts the quantity and quality of economically important crops. Recently, environmentally acceptable mitigating practices have been required for sustainable agriculture, restricting the use of chemical fertilizers in agricultural areas. Herein, the biological control of Zucchini yellow mosaic virus (ZYMV) in cucumber (Cucumis sativus L.) plants using arbuscular mycorrhizal (AM) fungi was investigated. Compared to control plants, ZYMV-infected plants displayed high disease incidence (DI) and severity (DS) with various symptoms, including severe yellow mosaic, mottling and green blisters of leaves. However, AM fungal inoculation exhibited 50% inhibition for these symptoms and limited DS to 26% as compared to non-colonized ones. The detection of ZYMV by the Enzyme-Linked Immunosorbent Assay technique exhibited a significant reduction in AM-inoculated plants (5.23-fold) compared with non-colonized ones. Besides, mycorrhizal root colonization (F%) was slightly reduced by ZYMV infection. ZYMV infection decreased all growth parameters and pigment fractions and increased the malondialdehyde (MDA) content, however, these parameters were significantly enhanced and the MDA content was decreased by AM fungal colonization. Also, the protein, proline and antioxidant enzymes (POX and CAT) were increased with ZYMV infection with more enhancements due to AM root colonization. Remarkably, defence pathogenesis-related (PR) genes such as PR-a, PR-b, and PR-10 were quickly expressed in response to AM treatment. Our findings demonstrated the beneficial function of AM fungi in triggering the plant defence against ZYMV as they caused systemic acquired resistance in cucumber plants and supported their potential use in the soil as an environment-friendly method of hindering the spread of pathogenic microorganisms sustainably.

Overexpression of cucumber CYP82D47 enhances resistance to powdery mildew and Fusarium oxysporum f. sp. cucumerinum.

Cytochrome P450s are a large family of protein-encoding genes in plant genomes, many of which have not yet been comprehensively characterized. Here, a novel P450 gene, CYP82D47, was isolated and functionally characterized from cucumber (Cucumis sativus L.). Quantitative real-time reverse-transcription polymerase chain reaction analysis revealed that CYP82D47 expression was triggered by salicylic acid (SA) and ethephon (ETH). Expression analysis revealed a correlation between CYP82D47 transcript levels and plant defense responses against powdery mildew (PM) and Fusarium oxysporum f. sp. cucumerinum (Foc). Although no significant differences were observed in disease resistance between CYP82D47-RNAi and wild-type cucumber, overexpression (OE) of CYP82D47 enhanced PM and Foc resistance in cucumber. Furthermore, the expression levels of SA-related genes (PR1, PR2, PR4, and PR5) increased in CYP82D47-overexpressing plants 7days post fungal inoculation. The levels of ETH-related genes (EIN3 and EBF2) were similarly upregulated. The observed enhanced resistance was associated with the upregulation of SA/ETH-signaling-dependent defense genes. These findings indicate the crucial role of CYP82D47 in pathogen defense in cucumber. CYP82D47-overexpressing cucumber plants exhibited heightened susceptibility to both diseases. The study results offer important insights that could aid in the development of disease-resistant cucumber cultivars and elucidate the molecular mechanisms associated with the functions of CYP82D47.

Methods for seedling identification of cucumber resistance to powdery mildew and its effect on the growth of cucumber seedlings

Methods for seedling identification of cucumber resistance to powdery mildew and its effect on the growth of cucumber seedlings

CsMLO8/11 are required for full susceptibility of cucumber stem to powdery mildew and interact with CsCRK2 and CsRbohD.

Powdery mildew (PM) is one of the most destructive diseases that threaten cucumber production globally. Efficient breeding of novel PM-resistant cultivars will require a robust understanding of the molecular mechanisms of cucumber resistance against PM. Using a genome-wide association study, we detected a locus significantly correlated with PM resistance in cucumber stem, pm-s5.1. A 1449-bp insertion in the CsMLO8 coding region at the pm-s5.1 locus resulted in enhanced stem PM resistance. Knockout mutants of CsMLO8 and CsMLO11 generated by CRISPR/Cas9 both showed improved PM resistance in the stem, hypocotyl, and leaves, and the double mutant mlo8mlo11 displayed even stronger resistance. We found that reactive oxygen species (ROS) accumulation was higher in the stem of these mutants. Protein interaction assays suggested that CsMLO8 and CsMLO11 could physically interact with CsRbohD and CsCRK2, respectively. Further, we showed that CsMLO8 and CsCRK2 competitively interact with the C-terminus of CsRbohD to affect CsCRK2-CsRbohD module-mediated ROS production during PM defense. These findings provide new insights into the understanding of CsMLO proteins during PM defense responses.

Endogenous heme positively contributes to ALA-enhanced salt stress resistance in cucumber

In order to investigate the time-dependent rule of heme that is implicated in the salt resistance of cucumber seedlings, it is imperative to conduct a thorough investigation.Hence, this study endeavors subject cucumber (Cucumis sativus) plants to treatments involving exogenous heme synthesis inhibitors (DPD, 0.2 mmol∙L−1), heme scavengers (Hx, 0.4 μg∙L−1), and ALA (25 mg∙L−1) to investigate the response and regulatory mechanisms, photosynthetic capacity, and antioxidant capacity under salt stress (50 mmol∙L−1). The findings revealed that the concentration of heme in cucumber leaves increased by 55.8%, 107.8%, and 166.9%, respectively, on the 3rd, 5th, and 7th days under salt stress, in comparison to the level of control. The malondialdehyde (MDA) content in the leaves of seedlings under single salt stress increased by 16.3%, 27.6%, and 11.8%, respectively, and increased by 30.9%, 104.1%, and 89.4% in the roots. Proline content in leaves and roots of seedlings gradually accumulated under single salt stress. Furthermore, the CAT activity increased by 2.8% in comparison with the normal level on the 7th day of salt stresss. On the seventh day of treatment, the total chlorophyll content experienced a decrease of 36.8% in comparison to the control level. Correspondingly, the photosynthetic capacity of the leaves was also inhibited. On the 7th day of NaCl+DPD+Hx treatment, the concentrations of MDA in leaves and MDA in roots exhibited an increase by 39.6% and 17.3%, respectively, the concentrations of ChlT, Y(II), Pn, E, and gs were observed to decrease by 11.0%, 17.6%, 38.3%, 23.8%, and 44.0%, respectively, in comparison to a single salt treatment. Nonetheless, subsequent to the addition of exogenous ALA under stress, the concentrations of MDA and proline decreased, the activity of antioxidant enzymes remained stable, the concentrations of heme and chlorophyll increased, and the photosynthetic capacity augmented.In summary, heme is positively involved in its own defense mechanism and exogenous ALA-induced salt resistance enhancement in cucumber seedlings.