scholarly journals Pseudomonas fluorescens DN16 Enhances Cucumber Defense Responses Against the Necrotrophic Pathogen Botrytis cinerea by Regulating Thermospermine Catabolism

2021 ◽  
Vol 12 ◽  
Author(s):  
Lin Zhu ◽  
Nana Qian ◽  
Yujun Sun ◽  
Xiaoming Lu ◽  
Haiming Duan ◽  
...  

Plants can naturally interact with beneficial rhizobacteria to mediate defense responses against foliar pathogen infection. However, the mechanisms of rhizobacteria-mediated defense enhancement remain rarely clear. In this study, beneficial rhizobacterial strain Pseudomonas fluorescens DN16 greatly increased the resistance of cucumber plants against Botrytis cinerea infection. RNA-sequencing analyses showed that several polyamine-associated genes including a thermospermine (TSpm) synthase gene (CsACL5) and polyamine catabolic genes (CsPAO1, CsPAO5, and CsCuAO1) were notably induced by DN16. The associations of TSpm metabolic pathways with the DN16-mediated cucumber defense responses were further investigated. The inoculated plants exhibited the increased leaf TSpm levels compared with the controls. Accordantly, overexpression of CsACL5 in cucumber plants markedly increased leaf TSpm levels and enhanced defense against B. cinerea infection. The functions of TSpm catabolism in the DN16-mediated defense responses of cucumber plants to B. cinerea were further investigated by pharmacological approaches. Upon exposure to pathogen infection, the changes of leaf TSpm levels were positively related to the enhanced activities of polyamine catabolic enzymes including polyamine oxidases (PAOs) and copper amine oxidases (CuAOs), which paralleled the transcription of several defense-related genes such as pathogenesis-related protein 1 (CsPR1) and defensin-like protein 1 (CsDLP1). However, the inhibited activities of polyamine catabolic enzymes abolished the DN16-induced cucumber defense against B. cinerea infection. This was in line with the impaired expression of defense-related genes in the inoculated plants challenged by B. cinerea. Collectively, our findings unraveled a pivotal role of TSpm catabolism in the regulation of the rhizobacteria-primed defense states by mediating the immune responses in cucumber plants after B. cinerea infection.

2021 ◽  
Vol 12 ◽  
Author(s):  
Beibei Li ◽  
Ruolin Wang ◽  
Shiya Wang ◽  
Jiang Zhang ◽  
Ling Chang

Cytokinins (CKs) can modulate plant immunity to various pathogens, but how CKs are involved in plant defense responses to the necrotrophic pathogen Botrytis cinerea is still unknown. Here, we found that B. cinerea infection induced transcriptional changes in multiple genes involved in the biosynthesis, degradation, and signaling of CKs, as well as their contents, in pathogen-infected Arabidopsis leaves. Among the CKs, the gene expression of CYTOKININ OXIDASE/DEHYDROGENASE 5 (CKX5) was remarkably induced in the local infected leaves and the distant leaves of the same plant without pathogen inoculation. Cis-zeatin (cZ) and its riboside (cZR) accumulated considerably in infected leaves, suggesting an important role of the cis-zeatin type of CKs in the plant response to B. cinerea. Cytokinin double-receptor mutants were more susceptible to B. cinerea infection, whereas an exogenous CK treatment enhanced the expression levels of defense-related genes and of jasmonic acid (JA) and ethylene (ET), but not salicylic acid (SA), resulting in higher resistance of Arabidopsis to B. cinerea. Investigation of CK responses to B. cinerea infection in the JA biosynthesis mutant, jar1-1, and ET-insensitive mutant, ein2-1, showed that CK signaling and levels of CKs, namely, those of isopentenyladenine (iP), isopentenyladenine riboside (iPR), and trans-zeatin (tZ), were enhanced in jar1-1-infected leaves. By contrast, reductions in iP, iPR, tZ, and tZ riboside (tZR) as well as cZR contents occurred in ein2-1-infected leaves, whose transcript levels of CK signaling genes were likewise differentially regulated. The Arabidopsis Response Regulator 5 (ARR5) gene was upregulated in infected leaves of ein2-1 whereas another type-A response regulator, ARR16, was significantly downregulated, suggesting the existence of a complex regulation of CK signaling via the ET pathway. Accumulation of the cis-zeatin type of CKs in B. cinerea-infected leaves depended on ET but not JA pathways. Collectively, our findings provide evidence that CK responds to B. cinerea infection in a variety of ways that are differently modulated by JA and ET pathways in Arabidopsis.


Genes ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 1007
Author(s):  
Divya Kattupalli ◽  
Asha Sreenivasan ◽  
Eppurathu Vasudevan Soniya

Black pepper (Piper nigrum L.) is a prominent spice that is an indispensable ingredient in cuisine and traditional medicine. Phytophthora capsici, the causative agent of footrot disease, causes a drastic constraint in P. nigrum cultivation and productivity. To counterattack various biotic and abiotic stresses, plants employ a broad array of mechanisms that includes the accumulation of pathogenesis-related (PR) proteins. Through a genome-wide survey, eleven PR-1 genes that belong to a CAP superfamily protein with a caveolin-binding motif (CBM) and a CAP-derived peptide (CAPE) were identified from P. nigrum. Despite the critical functional domains, PnPR-1 homologs differ in their signal peptide motifs and core amino acid composition in the functional protein domains. The conserved motifs of PnPR-1 proteins were identified using MEME. Most of the PnPR-1 proteins were basic in nature. Secondary and 3D structure analyses of the PnPR-1 proteins were also predicted, which may be linked to a functional role in P. nigrum. The GO and KEGG functional annotations predicted their function in the defense responses of plant-pathogen interactions. Furthermore, a transcriptome-assisted FPKM analysis revealed PnPR-1 genes mapped to the P. nigrum-P. capsici interaction pathway. An altered expression pattern was detected for PnPR-1 transcripts among which a significant upregulation was noted for basic PnPR-1 genes such as CL10113.C1 and Unigene17664. The drastic variation in the transcript levels of CL10113.C1 was further validated through qRT-PCR and it showed a significant upregulation in infected leaf samples compared with the control. A subsequent analysis revealed the structural details, phylogenetic relationships, conserved sequence motifs and critical cis-regulatory elements of PnPR-1 genes. This is the first genome-wide study that identified the role of PR-1 genes during P. nigrum-P. capsici interactions. The detailed in silico experimental analysis revealed the vital role of PnPR-1 genes in regulating the first layer of defense towards a P. capsici infection in Panniyur-1 plants.


2020 ◽  
Vol 21 (19) ◽  
pp. 7352
Author(s):  
Shoya Kitabayashi ◽  
Daiki Tsushima ◽  
Charith Raj Adkar-Purushothama ◽  
Teruo Sano

While the potato spindle tuber viroid (PSTVd) variant, PSTVd-Dahlia (PSTVd-D or PSTVd-Dwt) induces very mild symptoms in tomato cultivar ‘Rutgers’, PSTVd-Intermediate (PSTVd-I or PSTVd-Iwt) induces severe symptoms. These two variants differ by nine nucleotides, of which six mutations are located in the terminal left (TL) to the pathogenicity (P) domains. To evaluate the importance of mutations located in the TL to the P domains, ten types of point mutants were created by swapping the nucleotides between the two viroid variants. Bioassay in tomato plants demonstrated that two mutants created on PSTVd-Iwt at positions 42 and 64 resulted in symptom attenuation. Phenotypic and RT-qPCR analysis revealed that mutation at position 42 of PSTVd-Iwt significantly reduced disease severity and accumulation of the viroid, whereas mutation at position 64 showed a significant reduction in stunting when compared to the PSTVd-Iwt infected plant. RT-qPCR analysis on pathogenesis-related protein 1b1 and chalcone synthase genes showed a direct correlation with symptom severity whereas the expansin genes were down-regulated irrespective of the symptom severity. These results indicate that the nucleotides at positions 42 and 64 are in concert with the ones at positions 43, 310, and 311/312, which determines the slower and stable accumulation of PSTVd-D without eliciting excessive host defense responses thus contributing in the attenuation of disease symptom.


1999 ◽  
Vol 12 (6) ◽  
pp. 479-489 ◽  
Author(s):  
Sze-Chung Clive Lo ◽  
John D. Hipskind ◽  
Ralph L. Nicholson

A sorghum cDNA clone was isolated by differential screening of a cDNA library prepared from mesocotyls (cultivar DK18) inoculated with fungal pathogens. The deduced translation product shows sequence similarity to a family of intracellular pathogenesis-related proteins (PR-10) with a potential ribonuclease function. We studied the accumulation of PR-10 and chalcone synthase (CHS) transcripts in mesocotyls following inoculation with Cochliobolus heterostrophus or Colletotrichum sublineolum. CHS is involved in phytoalexin synthesis in sorghum. Coordinate expression of PR-10 and CHS genes was localized in the area of inoculation along with the accumulation of phytoalexins. C. heterostrophus is a nonpathogen of sorghum and cytological studies indicated that cultivar DK18 is resistant to C. sublineolum, a sorghum pathogen. We demonstrated that the two fungi triggered different time courses of plant defense reactions. Inoculation with C. heterostrophus resulted in rapid accumulation of PR-10 and CHS transcripts after appressoria had become mature. Accumulation of these transcripts was delayed in plants inoculated with C. sublineolum until penetration of host tissue had been completed and infection vesicles had formed. Results suggest that different recognition events are involved in the expression of resistance to the two fungi used or that C. sublineolum suppresses the nonspecific induction of defense responses.


2015 ◽  
Vol 28 (10) ◽  
pp. 1117-1129 ◽  
Author(s):  
Charlotte Gruau ◽  
Patricia Trotel-Aziz ◽  
Sandra Villaume ◽  
Fanja Rabenoelina ◽  
Christophe Clément ◽  
...  

Although induced systemic resistance (ISR) is well-documented in the context of plant–beneficial bacteria interactions, knowledge about the local and systemic molecular and biochemical defense responses before or upon pathogen infection in grapevine is very scarce. In this study, we first investigated the capacity of grapevine plants to express immune responses at both above- and below-ground levels upon interaction with a beneficial bacterium, Pseudomonas fluorescens PTA-CT2. We then explored whether the extent of priming state could contribute to the PTA-CT2-induced ISR in Botrytis cinerea–infected leaves. Our data provide evidence that this bacterium colonized grapevine roots but not the above-ground plant parts and altered the plant phenotype that displayed multiple defense responses both locally and systemically. The grapevine roots and leaves exhibited distinct patterns of defense-related gene expression during root colonization by PTA-CT2. Roots responded faster than leaves and some responses were more strongly upregulated in roots than in leaves and vice versa for other genes. These responses appear to be associated with some induction of cell death in roots and a transient expression of HSR, a hypersensitive response-related gene in both local (roots) and systemic (leaves) tissues. However, stilbenic phytoalexin patterns followed opposite trends in roots compared with leaves but no phytoalexin was exuded during plant-bacterium interaction, suggesting that roots could play an important role in the transfer of metabolites contributing to immune response at the systemic level. Unexpectedly, in B. cinerea–infected leaves PTA-CT2-mediated ISR was accompanied in large part by a downregulation of different defense-related genes, including HSR. Only phytoalexins and glutathion-S-transferase 1 transcripts were upregulated, while the expression of anthocyanin biosynthetic genes was maintained at a higher level than the control. This suggests that decreased expression of HSR, as a marker of cell death, and activation of secondary metabolism pathways could be responsible for a reduced B. cinerea colonization capacity in bacterized plants.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Kai Bi ◽  
Loredana Scalschi ◽  
Namrata Jaiswal ◽  
Tesfaye Mengiste ◽  
Renana Fried ◽  
...  

AbstractCrh proteins catalyze crosslinking of chitin and glucan polymers in fungal cell walls. Here, we show that the BcCrh1 protein from the phytopathogenic fungus Botrytis cinerea acts as a cytoplasmic effector and elicitor of plant defense. BcCrh1 is localized in vacuoles and the endoplasmic reticulum during saprophytic growth. However, upon plant infection, the protein accumulates in infection cushions; it is then secreted to the apoplast and translocated into plant cells, where it induces cell death and defense responses. Two regions of 53 and 35 amino acids are sufficient for protein uptake and cell death induction, respectively. BcCrh1 mutant variants that are unable to dimerize lack transglycosylation activity, but are still able to induce plant cell death. Furthermore, Arabidopsis lines expressing the bccrh1 gene exhibit reduced sensitivity to B. cinerea, suggesting a potential use of the BcCrh1 protein in plant immunization against this necrotrophic pathogen.


Plants ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 970
Author(s):  
Danai Gkizi ◽  
Eirini G. Poulaki ◽  
Sotirios E. Tjamos

Grapevine bunch rot, caused by Botrytis cinerea and Aspergillus carbonarius, causes important economic losses every year in grape production. In the present study, we examined the plant protective activity of the biological control agents, Paenibacillus alvei K165, Blastobotrys sp. FP12 and Arthrobacter sp. FP15 against B. cinerea and A. carbonarius on grapes. The in vitro experiments showed that strain K165 significantly reduced the growth of both fungi, while FP15 restricted the growth of A. carbonarius and FP12 was ineffective. Following the in vitro experiments, we conducted in planta experiments on grape berries. It was shown that K165, FP12 and FP15 reduced A. carbonarius rot severity by 81%, 57% and 37%, respectively, compared to the control, whereas, in the case of B. cinerea, the only protective treatment was that with K165, which reduced rot by 75%. The transcriptomic analysis of the genes encoding the pathogenesis-related proteins PR2, PR3, PR4 and PR5 indicates the activation of multiple defense responses involved in the biocontrol activity of the examined biocontrol agents.


2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Arjun Sham ◽  
Hibatullah Al-Ashram ◽  
Kenna Whitley ◽  
Rabah Iratni ◽  
Khaled A. El-Tarabily ◽  
...  

AbstractIn this study, we aimed to identify common genetic components during stress response responsible for crosstalk among stresses, and to determine the role of differentially expressed genes in Arabidopsis-Botrytis cinerea interaction. Of 1,554 B. cinerea up-regulated genes, 24%, 1.4% and 14% were induced by biotic, abiotic and hormonal treatments, respectively. About 18%, 2.5% and 22% of B. cinerea down-regulated genes were also repressed by the same stress groups. Our transcriptomic analysis indicates that plant responses to all tested stresses can be mediated by commonly regulated genes; and protein-protein interaction network confirms the cross-interaction between proteins regulated by these genes. Upon challenges to individual or multiple stress(es), accumulation of signaling molecules (e.g. hormones) plays a major role in the activation of downstream defense responses. In silico gene analyses enabled us to assess the involvement of RAP2.4 (related to AP2.4) in plant immunity. Arabidopsis RAP2.4 was repressed by B. cinerea, and its mutants enhanced resistance to the same pathogen. To the best of our knowledge, this is the first report demonstrating the role of RAP2.4 in plant defense against B. cinerea. This research can provide a basis for breeding programs to increase tolerance and improve yield performance in crops.


Author(s):  
Yihao Li ◽  
Kun Liu ◽  
Ganlu Tong ◽  
Chao Xi ◽  
Jin Liu ◽  
...  

Abstract Ethylene response factor (ERF) Group VII members generally function in regulating plant growth and development, abiotic stress response, and plant immunity in Arabidopsis. However, the detail regulatory mechanism by which Group VII ERFs mediate plant immune responses remains elusive. Here, we characterised ERF72, a member of the Group VII ERFs, as a positive regulator mediating resistance to the necrotrophic pathogen Botrytis cinerea. Compared with wild type (WT), erf72 mutant showed the lower camalexin contents and more susceptible to B. cinerea, while complementation of ERF72 in erf72 rescued susceptibility phenotypes. Moreover, overexpression of ERF72 in WT promoted camalexin biosynthesis and resistance to B. cinerea. Then, we identified camalexin biosynthesis genes PAD3 and CYP71A13, and transcription factor WRKY33 as target genes of ERF72. Furthermore, MPK3 and MPK6 phosphorylate ERF72 at Ser151 to improve its transactivation activity, camalexin contents and resistance to B. cinerea. These findings highlight the role of ERF72 in coordinating the camalexin biosynthesis via directly regulating the expression of camalexin biosynthetic genes and indirectly by targeting WRKK33 in plant immunity.


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