Resistance In Cucumber Research Articles

CsPM5.2, a phosphate transporter protein-like gene, promotes powdery mildew resistance in cucumber.

Powdery mildew (PM) is one of the most serious fungal diseases affecting cucumbers (Cucumis sativus L.). The mechanism of PM resistance in cucumber is intricate and remains fragmentary as it is controlled by several genes. In this study, we detected the major-effect Quantitative Trait Locus (QTL), PM5.2, involved in PM resistance by QTL mapping. Through fine mapping, the dominant PM resistance gene, CsPM5.2, was cloned and its function was confirmed by transgenic complementation and natural variation identification. In cultivar 9930, a dysfunctional CsPM5.2 mutant resulted from a single nucleotide polymorphism in the coding region and endowed susceptibility to PM. CsPM5.2 encodes a phosphate transporter-like protein PHO1; H3. The expression of CsPM5.2 is ubiquitous and induced by the PM pathogen. In cucumber, both CsPM5.2 and Cspm5.1 (Csmlo1) are required for PM resistance. Transcriptome analysis suggested that the salicylic acid (SA) pathway may play an important role in CsPM5.2-mediated PM resistance. Our findings help parse the mechanisms of PM resistance and provide strategies for breeding PM-resistant cucumber cultivars.

A single-nucleotide substitution in the leucine-rich-repeat-only gene CsLRR1 confers powdery mildew resistance in cucumber

A single-nucleotide substitution in the leucine-rich-repeat-only gene CsLRR1 confers powdery mildew resistance in cucumber

β-Aminobutyric Acid and Powdery Mildew Infection Enhanced the Activation of Defense-Related Genes and Salicylic Acid in Cucumber (Cucumis sativus L.).

Powdery mildew disease, caused by Sphaerotheca fusca, is a major disease affecting cucumbers cultivated in greenhouses. This study was conducted to find defense genes induced by β-aminobutyric acid (BABA) and powdery mildew in cucumber. Disease severities of 25% and 5% were exhibited by the 2000 and 5000 mg/L BABA-treated cucumber, respectively. BABA did not affect the spore germination of the powdery mildew pathogen, showing that BABA is not an antifungal agent against the pathogen. In quantitative real-time PCR analysis, BABA-treated cucumber upregulated the transcriptional levels of the defense genes CsPAL, CsPR3, CsPR1, CsLOX1, CsLOX23, Cs LecRK6.1, CsWRKY20, and Cupi4 in cucumber to maximum levels at 48 h, whereas CsLecRK6.1 reached maximum expression after 24 h, and further, salicylic acid (SA) levels were significantly increased in BABA-treated cucumber plants. In addition, the cucumber infected with powdery mildew underwent a 1.6- to 47.3-fold enhancement in the defense genes PAL, PR3, PR1, Lox1, Lox 23, LecRK6.1, WRKY20, and Cupi4 compared to heathy cucumber. These results suggest that the BABA-induced defense response is associated with SA signaling pathway-dependent systemic acquired resistance (SAR) in cucumber, which is involved in plant resistance mechanisms.

Silencing the CsSnRK2.11 Gene Decreases Drought Tolerance of Cucumis sativus L.

Drought stress restricts vegetable growth, and abscisic acid plays an important role in its regulation. Sucrose non-fermenting1-related protein kinase 2 (SnRK2) is a key enzyme in regulating ABA signal transduction in plants, and it plays a significant role in response to multiple abiotic stresses. Our previous experiments demonstrated that the SnRK2.11 gene exhibits a significant response to drought stress in cucumbers. To further investigate the function of SnRK2.11 under drought stress, we used VIGS (virus-induced gene silencing) technology to silence this gene and conducted RNA-seq analysis. The SnRK2.11-silencing plants displayed increased sensitivity to drought stress, which led to stunted growth and increased wilting speed. Moreover, various physiological parameters related to photosynthesis, chlorophyll fluorescence, leaf water content, chlorophyll content, and antioxidant enzyme activity were significantly reduced. The intercellular CO2 concentration, non-photochemical burst coefficient, and malondialdehyde and proline content were significantly increased. RNA-seq analysis identified 534 differentially expressed genes (DEGs): 311 were upregulated and 223 were downregulated. GO functional annotation analysis indicated that these DEGs were significantly enriched for molecular functions related to host cells, enzyme activity, and stress responses. KEGG pathway enrichment analysis further revealed that these DEGs were significantly enriched in phytohormone signalling, MAPK signalling, and carotenoid biosynthesis pathways, all of which were associated with abscisic acid. This study used VIGS technology and transcriptome data to investigate the role of CsSnRK2.11 under drought stress, offering valuable insights into the mechanism of the SnRK2 gene in enhancing drought resistance in cucumbers.

Nano-selenium foliar intervention-induced resistance of cucumber to Botrytis cinerea by activating jasmonic acid biosynthesis and regulating phenolic acid and cucurbitacin.

Botrytis cinerea is the primary disease affecting cucumber production. It can be managed by applying pesticides and cultivating disease-resistant cucumber strains. However, challenges, such as drug resistance in pathogenic bacteria and changes in physiological strains, are obstacles in the effective management of B. cinerea. Nano-selenium (Nano-Se) has potential in enhancing crop resistance to biological stress, but the exact mechanism for boosting disease resistance remains unclear. Here, we used metabolomics and transcriptomics to examine how Nano-Se, as an immune activator, induces plant resistance. Compared with the control group, the application of 10.0 mg/L Nano-Se on the cucumber plant's leaf surface resulted in increased levels of chlorophyll, catalase (10.2%), glutathione (326.6%), glutathione peroxidase (52.2%), cucurbitacin (41.40%), and metabolites associated with the phenylpropane synthesis pathway, as well as the total antioxidant capacity (21.3%). Additionally, the expression levels of jasmonic acid (14.8 times) and related synthetic genes, namely LOX (264.1%), LOX4 (224.1%), and AOC2 (309.2%), were up-regulated. A transcription analysis revealed that the CsaV3_4G002860 gene was up-regulated in the KEGG enrichment pathway in response to B. cinerea infection following the 10.0 mg/L Nano-Se treatment. In conclusion, the activation of the phenylpropane biosynthesis and branched-chain fatty acid pathways by Nano-Se promotes the accumulation of jasmonic acid and cucurbitacin in cucumber plants. This enhancement enables the plants to exhibit resistance against B. cinerea infections. Additionally, this study identified a potential candidate gene for cucumber resistance to B. cinerea induced by Nano-Se, thereby laying a theoretical foundation for further research in this area. © 2023 Society of Chemical Industry.

Characterization of caffeoyl shikimate esterase gene family identifies CsCSE5 as a positive regulator of Podosphaera xanthii and Corynespora cassiicola pathogen resistance in cucumber.

CsCSE genes might be involved in the tolerance of cucumber to pathogens. Silencing of the CsCSE5 gene resulted in attenuated resistance of cucumber to Podosphaera xanthii and Corynespora cassiicola. Caffeoyl shikimate esterase (CSE), a key enzyme in the lignin biosynthetic pathway, has recently been characterized to play a key role in defense against pathogenic infection in plants. However, a systematic analysis of the CSE gene family in cucumber (Cucumis sativus) has not yet been conducted. Here, we identified eight CsCSE genes from the cucumber genome via bioinformatic analyses, and these genes were unevenly distributed on chromosomes 1, 3, 4, and 5. Results from multiple sequence alignment indicated that the CsCSE proteins had domains required for CSE activity. Phylogenetic analysis of gene structure and protein motifs revealed the conservation and diversity of the CsCSE gene family. Collinearity analysis showed that CsCSE genes had high homology with CSE genes in wax gourd (Benincasa hispida). Cis-acting element analysis of the promoters suggested that CsCSE genes might play important roles in growth, development, and stress tolerance. Expression pattern analysis indicated that CsCSE5 might be involved in regulating the resistance of cucumber to pathogens. Functional verification data confirmed that CsCSE5 positively regulates the resistance of cucumber to powdery mildew pathogen Podosphaera xanthii and target leaf spot pathogen Corynespora cassiicola. The results of our study provide information that will aid the genetic improvement of resistant cucumber varieties.

The exocyst subunit CsExo70B promotes both fruit length and disease resistance via regulating receptor kinase abundance at plasma membrane in cucumber.

Plant defence against pathogens generally occurs at the expense of growth and yield. Uncoupling the inverse relationship between growth and defence is of great importance for crop breeding, while the underlying genes and regulatory mechanisms remain largely elusive. The exocytosis complex was shown to play an important role in the trafficking of receptor kinases (RKs) to the plasma membrane (PM). Here, we found a Cucumis sativus exocytosis subunit Exo70B (CsExo70B) regulates the abundance of both development and defence RKs at the PM to promote fruit elongation and disease resistance in cucumber. Knockout of CsExo70B resulted in shorter fruit and susceptibility to pathogens. Mechanistically, CsExo70B associates with the developmental RK CsERECTA, which promotes fruit longitudinal growth in cucumber, and contributes to its accumulation at the PM. On the other side, CsExo70B confers to the spectrum resistance to pathogens in cucumber via a similar regulatory module of defence RKs. Moreover, CsExo70B overexpression lines showed an increased fruit yield as well as disease resistance. Collectively, our work reveals a regulatory mechanism that CsExo70B promotes both fruit elongation and disease resistance by maintaining appropriate RK levels at the PM and thus provides a possible strategy for superior cucumber breeding with high yield and robust pathogen resistance.

Relative biochemical and physiological docking of cucumber varieties for supporting innate immunity against Podosphaera xanthii

Powdery mildew in cucumber is caused by the Podosphaera xanthii. No strategy for improving disease resistance can be successful in the absence of thorough insights into the physiological and biochemical responses of cucumber plants to powdery mildew. Therefore, a field experiment was executed to evaluate five commercial cucumber varieties (V1: Dynasty, V2: Long green, V3:Desi Kheera, V4:Thamin II, V5:Cucumber 363) for their inherent immunity to powdery mildew. Upon inoculating cucumber plants with Podosphaera xanthii, we noted differential responses among the varieties. Compared to other varieties, V1 and V2 showed higher values (P ≤ 0.05) for chlorophyll-a under control and pathogen-attacked plants respectively. The minimum value of anthocyanin content (−53.73%) was recorded in V3 as compared to other varieties post pathogen infection. All pathogen-infected cucumber varieties showed a considerable (P ≤ 0.05) loss in flavonoid content except V2. The maximum destruction for Phenolics under powdery mildew (179%) were recorded in V4, whereas V1 exhibited maximum phenolic content under control conditions. In pathogen-infected plants, the minimum AsA was recorded in V5 as compared to all other varieties. Pathogen invasion impacted significantly (P ≤ 0.05) the activity of superoxide dismutase (SOD). Besides, cucumber plants after pathogen inoculation resulted in a considerable (P ≤ 0.05) increase of peroxidase (POD) activity in V1 (5.02%), V2 (7.5%), and V3 (11%) in contrast to V4. Our results confirmed that cucumber varieties perform differently, which was brought on by distinct metabolic and physiological modifications that have an impact on growth and development. The changes in different attributes were correlated with cucumber resistance against powdery mildew. The results would help us fully harness the potential of these varieties to trigger disease management initiatives and defense responses.

Transcriptome Analyses Revealed the Wax and Phenylpropanoid Biosynthesis Pathways Related to Disease Resistance in Rootstock-Grafted Cucumber.

Cucumbers (Cucumis sativus L.) are a global popular vegetable and are widely planted worldwide. However, cucumbers are susceptible to various infectious diseases such as Fusarium and Verticillium wilt, downy and powdery mildew, and bacterial soft rot, which results in substantial economic losses. Grafting is an effective approach widely used to control these diseases. The present study investigated the role of wax and the phenylpropanoid biosynthesis pathway in black-seed pumpkin rootstock-grafted cucumbers. Our results showed that grafted cucumbers had a significantly higher cuticular wax contents on the fruit surface than that of self-rooted cucumbers at all stages observed. A total of 1132 differently expressed genes (DEGs) were detected in grafted cucumbers compared with self-rooted cucumbers. Pathway enrichment analysis revealed that phenylpropanoid biosynthesis, phenylalanine metabolism, plant circadian rhythm, zeatin biosynthesis, and diterpenoid biosynthesis were significantly enriched. In this study, 1 and 13 genes involved in wax biosynthesis and the phenylpropanoid biosynthesis pathway, respectively, were up-regulated in grafted cucumbers. Our data indicated that the up-regulated genes in the wax and phenylpropanoid biosynthesis pathways may contribute to disease resistance in rootstock-grafted cucumbers, which provides promising targets for enhancing disease resistance in cucumbers by genetic manipulation.

Effects of Rhizosphere Microbial Communities on Cucumber Fusarium wilt Disease Suppression.

Cucumber Fusarium wilt is a worldwide soil-borne disease that seriously restricts the yield and quality of cucumber. The rhizosphere soil microbiome, as the first line of defense against pathogens invading plant roots, plays a key role in rhizosphere immune formation and function. The purpose of this study was to reveal the key microecological factors and dominant microbial flora affecting cucumber resistance and susceptibility to Fusarium wilt by analyzing the physical and chemical properties and microbial flora of rhizosphere soil with different degrees of susceptibility and resistance to cucumber Fusarium wilt, thereby laying a foundation to establish cucumber resistance to the Fusarium wilt rhizosphere core microbiome. Firstly, Illumina Miseq sequencing technology was used to evaluate the physical and chemical properties and microbial groups of cucumber rhizosphere soil at different health levels, and the key environmental factors and microbial factors related to cucumber Fusarium wilt were screened out. Subsequently, PICRUSt2 and FUNGuild were used to predict the functions of rhizosphere bacteria and fungi. Combined with functional analysis, the possible interactions among soil physical and chemical properties, cucumber rhizosphere microorganisms, and Fusarium wilt were summarized. The results showed that the available potassium content in the rhizosphere soil of healthy cucumber decreased by 10.37% and 0.56%, respectively, compared with the rhizosphere soil of severely susceptible cucumber and mildly susceptible cucumber. Exchangeable calcium content increased by 25.55% and 5.39%; the α diversity Chao1 index of bacteria and fungi in the rhizosphere soil of healthy cucumber was significantly lower than that in the rhizosphere soil of seriously infected cucumber, and the MBC content of its physical and chemical properties was also significantly lower than that in the rhizosphere soil of seriously infected cucumber. There was no significant difference in the Shannon and Simpson diversity indexes between healthy cucumber rhizosphere soil and seriously infected cucumber rhizosphere soil. The results of the β diversity analysis showed that the bacterial and fungal community structure of healthy cucumber rhizosphere soil was significantly different from that of severely and mildly infected cucumber rhizosphere soil. At the genus level, through statistical analysis, LEfSe analysis, and RDA analysis, the key bacterial and fungal genera with potential biomarker values were screened out as SHA_26, Subgroup_22, MND1, Aeromicrobium, TM7a, Pseudorhodoplanes, Kocuria, Chaetomium, Fusarium, Olpidium, and Scopulariopsis, respectively. The bacteria SHA_26, Subgroup_22, and MND1 related to cucumber Fusarium wilt inhibition belong to Chloroflexi, Acidobacteriota, and Proteobacteria, respectively. Chaetomiacea belongs to Sordariomycates. The results of functional prediction showed that changes to the KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway in the bacterial microbiota were concentrated in tetracycline biosynthesis, selenocompound metabolism, lipopolysaccharide biosynthesis, etc., which were mainly involved in the metabolism of terpenoids and polyketides, energy metabolism, metabolism of other amino acids, glycan biosynthesis and metabolism, lipid metabolism, cell growth and death, transcription, metabolism of cofactors and vitamins, and biosynthesis of other secondary metabolites. The difference in fungi was mainly dung saprotroph-ectomycorrhizal-soil saprotroph-wood saprotroph. Through the correlation analysis and functional predictions of the key environmental factors, microbial flora, and cucumber health index in cucumber rhizosphere soil, we determined that the inhibition of cucumber Fusarium wilt was a synergistic effect of environmental factors and microbial flora, and a model diagram was drawn to briefly explain its mechanism. This work will provide a basis for the biological control of cucumber Fusarium wilt in the future.

Genetics, resistance mechanism, and breeding of powdery mildew resistance in cucumbers (Cucumis sativus L.)

Cucumber is an important vegetable worldwide, and powdery mildew (PM) is a common and serious disease of cucumbers. Breeding disease-resistant cucumber varieties is the most advantageous strategy to control this disease. In recent years, exploration and identification of cucumber PM resistance genes have achieved great advancement, and many genes have been cloned and verified using different methods. However, the resistance mechanism of cucumber PM is still unclear, and many ambiguities need to be elucidated urgently. In this review, we summarized the research advances in PM resistance in cucumbers, including genetic analysis, quantitative trait locus mapping, map-based cloning, transcriptomics, mlo-mediated PM resistance, and mining of noncoding RNAs involved in resistance. Finally, the research directions and the problems that need to be solved in the future were discussed.

Development of powdery mildew (Podosphaera xanthii) resistance in cucumber (Cucumis sativus) using CRISPR/Cas9-mediated mutagenesis of CsaMLO8.

Powdery mildew (PM) diseases may severely limit the production of various crops, including members of to the Cucurbitaceae family. Successful PM infection relies on the Mildew Resistance Locus O (MLO) plant gene family, which encodes susceptibility (S) factors essential for fungus penetration into the host cell. In cucumber, natural mutations in CsaMLO8 confer resistance to the PM pathogen Podosphaera xanthii. Here, we used CRISPR/Cas9-mediated mutagenesis to generate PM resistance in the susceptible cucumber cv. 'Ilan'. Two transgene-free Csamlo8 CRISPR mutant lines (Csamlo-cr-1 and Csamlo-cr-2) were isolated, the first with a 5 bp deletion in exon 1, and the second harboring a 1280 bp deletion and 10 bp insertion between exons 1 and 5. Both lines showed high resistance to PM under semi-commercial growth conditions in the summer growing seasons of 2019 and 2021. These results provide the basis for generating transgene-free powdery mildew resistance in cucumber in any genetic background. This method can directly be employed on commercial cultivars and hybrid parental lines, and thereby substantially shorten and simplify the breeding process for PM resistance in cucumber.

SnRK1 signaling regulates cucumber growth and resistance to Corynespora cassiicola

Energy metabolism is one of the key factors determining the growth and development of plants and the response to biotic and abiotic stresses. Sucrose non-fermentation 1 related protein kinase 1 (SnRK1) is an important energy-sensitive regulator that plays a key role in the overall control of carbohydrate metabolism. However, little is known about the function of SnRK1 in cucumber. In this study, metformin (an SnRK1 activator) and trehalose (an SnRK1 inhibitor) were used to investigate the role of SnRK1 signaling in cucumber. The results showed that SnRK1 activation could inhibit the growth of cucumber, slow down the net photosynthetic rate (Pn), reduce the contents of photosynthetic pigments and soluble sugars, and suppress the expression of genes related to sucrose metabolism. By contrast, SnRK1 inhibition yielded opposite results. Furthermore, SnRK1 activation and CsSnRK1 over-expression improved cucumber resistance to Corynespora cassiicola. While, SnRK1 inhibition and CsSnRK1 silencing reduced the resistance of cucumber to C. cassiicola. The results indicated that CsSnRK1 gene can positively regulate the resistance of cucumber to C. cassiicola. We conclude that CsSnRK1 signaling plays an important role in balancing the growth and immune response of cucumber. These results can be applied to the improvement of disease-resistant cucumber varieties.

A bHLH transcription factor, CsSPT, regulates high-temperature resistance in cucumber

High-temperature stress threatens the growth and yield of crops. Basic helix-loop-helix (bHLH) transcription factors (TFs) have been shown to play important roles in regulating high-temperature resistance in plants. However, the bHLH TFs responsible for high-temperature tolerance in cucumbers have not been identified. We used transcriptome profiling to screen the high temperature-responsive candidate bHLH TFs in cucumber. Here, we found that the expression of 75 CsbHLH genes was altered under high-temperature stress. The expression of the CsSPT gene was induced by high temperatures in TT (Thermotolerant) cucumber plants. However, the Csspt mutant plants obtained by the CRISPR-Cas9 system showed severe thermosensitive symptoms, including wilted leaves with brown margins and reduced root density and cell activity. The Csspt mutant plants also exhibited elevated H2O2 levels and down-regulated photosystem-related genes under normal conditions. Furthermore, there were high relative electrolytic leakage (REC), malondialdehyde (MDA), glutathione (GSH), and superoxide radical (O2·−) levels in the Csspt mutant plants, with decreased Proline content after the high-temperature treatment. Transcriptome analysis showed that the photosystem and chloroplast activities in Csspt mutant plants were extremely disrupted by the high-temperature stress compared with wild-type (WT) plants. Moreover, the plant hormone signal transduction, as well as MAPK and calcium signaling pathways were activated in Csspt mutant plants under high-temperature stress. The HSF and HSP family genes shared the same upregulated expression patterns in Csspt and WT plants under high-temperature conditions. However, most bHLH, NAC, and bZIP family genes were significantly down-regulated by heat in Csspt mutant plants. Thus, these results demonstrated that CsSPT regulated the high-temperature response by recruiting photosynthesis components, signaling pathway molecules, and transcription factors. Our results provide important insights into the heat response mechanism of CsSPT in cucumber and its potential as a target for breeding heat-resistant crops.

Salicylic acid functionalized zein for improving plant stress resistance and as a nanopesticide carrier with enhanced anti-photolysis ability

BackgroundThere is a serious global problem of salinization of arable land, causing large reduction in world food production. Use of plant hormones is an effective way to reduce damage caused to crops and salt stress.ResultsIn this study, PEI-EDA was modified with AM-zein and grafted with plant hormone SA (AM-zein-SA) and used as a nano-pesticide carrier to load emamectin benzoate (EB). The use of AM-zein-SA as a nano-pesticide carrier could reduce the damage caused by salt stress to crops. The structure of AM-zein-SA was characterized by FTIR, UV, fluorescence, Raman, and 1H NMR spectroscopic techniques. AM-zein-SA could effectively improve the resistance of EB to ultraviolet radiations, resistance of cucumber to salt stress, and the absorption of EB by plants. The experimental results showed that AM-zein-SA could effectively improve the anti-UV property of EB by 0.88 fold. When treated with 120 mmol NaCl, the germination rate of cucumber seeds under salt stress increased by 0.93 fold in presence of 6.25 mg/L carrier concentration. The POD and SOD activities increased by 0.50 and 1.21 fold, whereas the content of MDA decreased by 0.23 fold. In conclusion, AM-zein-SA nano-pesticide carrier could be used to improve the salt resistance of crops and the adhesion of pesticides to leaves.ConclusionAM-zein-SA, without undergoing any changes in its insecticidal activity, could simultaneously improve the salt stress resistance and salt stress germination rate of cucumber, reduce growth inhibition due to stress under high-concentration salt, and had a good effect on crops. In addition, EB@AM-zein-SA obviously improved the upward transmission rate of EB, as compared with EB. In this study, SA was grafted onto zein-based nano-pesticide carrier, which provided a green strategy to control plant diseases, insects, and pests while reducing salt stress on crops in saline-alkali soil.Graphical

The phloem lectin PP2-A1 enhances aphid resistance by affecting aphid behavior and maintaining ROS homeostasis in cucumber plants

Aphid (Aphis gossypii Glover) attack frequently results in a significant loss of output and deterioration of fruit quality in cucumber (Cucumis sativus L.). Phloem protein 2 (PP2) is conserved as a phloem lectin in plants, and few studies have been conducted on the regulatory mechanism of PP2. Based on our previous study of CsPP2-A1 in cucumber, to further investigate the biological function of CsPP2-A1, we compared the changes of selectivity, non-selectivity, colonization, reproductions of aphids, and the phenotype in wild type (WT), CsPP2-A1 overexpressing (CsPP2-A1-OE), and CsPP2-A1 interfering (CsPP2-A1-RNAi) cucumber plants after inoculation with aphids. We found that CsPP2-A1-OE cucumber plants generated resistance to aphids. The aphid colonization rate and number of reproductions of CsPP2-A1-OE cucumber plants were significantly lower than that of WT and CsPP2-A1-RNAi cucumber plants. Through Pearson's correlation and principal component analysis (PCA), it was found that CsPP2-A1 played a crucial role in the balance of reactive oxygen species (ROS) in plants. Overexpression of the CsPP2-A1 resulted in increased levels of antioxidant enzyme, eliminating ROS and preventing the damage by ROS in cucumber. Furthermore, nutritional imbalance for aphids and content of secondary metabolites were increased in overexpressed CsPP2-A1 cucumber plants, and thus preventing aphid attack. These together may improve cucumber resistance against aphids and the mechanism of CsPP2-A1 defense against aphids was preliminarily explored.

CRISPR/Cas9 based mlo-mediated resistance against Podosphaera xanthii in cucumber (Cucumis sativus L.).

Powdery mildews (PM) are common and severe pathogen groups that threaten plants, and PM resistance is complex and polygenic in cucumbers. Previously mlo-based resistance was reported in various plants, including cucumber, with generated loss-of CsaMLO function mutants. However, mlo-based resistance in cucumber is also complex and involves additional mechanisms such as hypersensitive response (HR) and papillae formation. For this reason, we focused on determining the mlo-based powdery mildew resistance mechanism in cucumber. CRISPR/Cas9 was used in the present study to generate loss-of-function mutants for CsaMLO1, CsaMLO8, and CsaMLO11 of PM susceptible ADR27 cucumber inbred lines and CsaMLO mutants were obtained and validated. Trypan Blue and DAB staining were performed to detect Podosphaera xanthii germination/penetration rates and accumulation of Reactive Oxygen Species (ROS). Our results indicate that PM-susceptibility associated CsaMLOs in cucumber are negative regulators in different defense mechanisms against powdery mildew at early and late stages of infection. Further, the experiment results indicated that CsaMLO8 mutation-based resistance was associated with the pre-invasive response, while CsaMLO1 and CsaMLO11 could be negative regulators in the post-invasive defense response in cucumber against P. xanthii. Although the loss-of CsaMLO8 function confers the highest penetration resistance, CsaMLO1 and CsaMLO11 double mutations could be potential candidates for HR-based resistance against PM pathogen in cucumber. These results highlighted the crucial role of CRISPR/Cas9 to develop PM resistant cucumber cultivars, possessing strong pre-invasive defense with CsaMLO8 or post-invasive with CsaMLO1/CsaMLO11 mutations.

Ultraviolet-B Irradiation Induces Resistance against Powdery Mildew in Cucumber (Cucumis sativus L.) through a Different Mechanism Than That of Heat Shock-Induced Resistance

Heat shock treatment (HST) and UV-B irradiation can reduce pathogen infection in crops. However, information on the mechanism of UV-B action is limited. Here, we investigated the mechanism of UV-B-induced resistance against powdery mildew in cucumber and compared it to that of heat-shock-induced resistance. We measured the percentage of leaf area showing disease symptoms and examined the expression levels of defense- and heat-shock-related genes across treatment groups. UV-B irradiation (intensity, 5 µW/cm2) for 4 h/d followed by pathogen inoculation reduced the appearance of powdery mildew by 21.17% compared with the control group. Unlike HST—which induces systemic resistance—UV-B irradiation induced local resistance in cucumber, as indicated by local changes in gene expression (Chi2 and ETR2). UV-B-treated plants inoculated with powdery mildew showed higher expression levels of Chi2, ETR2, and LOX6 than plants that were either treated with UV-B or inoculated. UV-B had no major effects on systemic acquired resistance or heat shock transcription factors, which are known to be affected by HST. Combined HST and UV-B had a strong synergistic effect in reducing powdery mildew in cucumber. Our results indicate that UV-B treatment likely operates through a different mechanism than HST in triggering cucumber resistance against powdery mildew infection.

Enhancement of cucumber resistance under salt stress by 2, 4-epibrassinolide lactones.

This study investigated the effects of exogenous 2, 4-epibrassinolide lactone (EBR) on the growth, photosynthetic pigments, antioxidant defense system, ion homeostasis, MAPK cascade and key genes of SOS signaling pathway of cucumber seedlings under salt stress using cucumber "Xinchun 4" as the test material. The experiment was set up with four treatments: foliar spraying of distilled water (CK), 50 mmol.L-1 NaCl (NaCl), 50 mmol.L-1 NaCl+foliar spray of 0.02 μmol.L-1 EBR (EBR+NaCl), and 50 mmol.L-1 NaCl+foliar spray of 24 μmol.L-1 Brassinazole (BRZ) (BRZ+NaCl). The results showed that EBR+NaCl treatment significantly increased plant height, above-ground fresh weight, total root length, total root surface area, average rhizome and photosynthetic pigment content compared to NaCl treatment. Meanwhile, compared with NaCl treatment, EBR+NaCl treatment significantly increased superoxide dismutase, catalase and ascorbate peroxidase (SOD, CAT and APX) activities, significantly promoted the accumulation of osmoregulatory substances (soluble sugars and proline), and thus effectively reduced malondialdehyde (MDA) content and relative electrical conductivity of cucumber leaves. Exogenous spraying of EBR also significantly reduced Na+/K+ under NaCl stress, effectively alleviating the toxic effects of Na+ ions. In addition, exogenous EBR induced the up-regulated expression of CsMAPK3, CsMAPK4, CsMAPK6 and CsMAPK9 genes in the MAPK cascade signaling pathway and CsSOS1, CsSOS2 and CsSOS3 genes in the SOS signaling pathway to enhance salt tolerance in cucumber under NaCl stress. Therefore, exogenous spraying EBR may effectively reduce the damage of salt stress on cucumber seedlings by improving antioxidant capacity, maintaining ion homeostasis and activating salt-tolerant related signaling pathways, which might promote the growth of cucumber seedlings and the establishment of root system morphology. This study provides a reference for EBR to improve the salt tolerance of cucumber.

Design, Synthesis, and Fungicidal Activity of Novel Plant Elicitors Based on a Diversity-Oriented Synthesis Strategy

The novel plant elicitors, 3-benzyl-5-[1-(2-oxo-4-phenyl-1-oxaspiro[4.5]dec-3-en-3-yl)ethylidene]-2-aminoimidazolin-4-one derivatives, were designed based on the diversity-oriented synthesis strategy and synthesized in four steps via the Knoevenagel condensation reaction as the key step. They were characterized by 1H NMR, 13C NMR, HR-ESI-MS, and X-ray diffraction. The position of the C═N bond of Z- and E-configuration compounds was determined by X-ray diffraction. The in vivo fungicidal activity evaluation revealed that most of these compounds exhibited remarkable activities (100%) against Pseudoperonospora cubensis at 400 μg/mL, among which compound 8e still exhibited excellent protective activity with a 50% inhibition rate at 0.1 μg/mL. Because the in vitro effect on tested phytopathogens was poor, the mechanism to induce the immune responses and reinforce the resistance of cucumber against Botrytis cinerea was studied. The results indicated that the compound 8e-mediated defense response against B. cinerea was based on the accumulation of pathogenesis-related proteins and cell wall reinforcement by callose deposition. Quantitative analysis of salicylic acid (SA) and jasmonic acid (JA) and the increased expression of induced resistance-related genes and the defense-associated phenylalanine ammonia lyase revealed that the immune response triggered by compound 8e was highly associated with the SA signaling pathway. Significant upregulation of JA-related genes Cs-AOS2 indicated that the JA signaling pathway was also influenced. It was also shown that the plants treated with compound 8e promoted primary root elongation, which resulted in enhanced plant growth. Most importantly, these compounds have completely new structures compared with the traditional plant elicitors. Further research of 8e-mediated plant disease resistance might have a great influence on the development of plant elicitors.