Phytophthora Capsici In Pepper Research Articles

A highly sensitive biotin-based probe for small RNA northern blot and its application in dissecting miRNA function in pepper.

Small RNAs play important roles in regulation of plant development and response to various stresses. Northern blot is an important technique in small RNA research. Isotope- and biotin- (or digoxigenin) labeled probes are frequently used in small RNA northern blot. However, isotope-based probe is limited by strict environmental regulation and availability in many places in the world while biotin-based probe is usually suffered from low sensitivity. In this study, we developed a T4 DNA polymerase-based method for incorporation of a cluster of 33 biotin-labeled C in small RNA probe (T4BC33 probe). T4BC33 probe reaches similar sensitivity as 32P-labeled probe in dot blot and small RNA northern blot experiments. Addition of locked nucleic acids in T4BC33 probe further enhanced its sensitivity in detecting low-abundance miRNAs. With newly developed northern blot method, expression of miR6027 and miR6149 family members was validated. Northern blot analysis also confirmed the successful application of virus-based miRNA silencing in pepper, knocking down accumulation of Can-miR6027a and Can-miR6149L. Importantly, further analysis showed that knocking-down Can-miR6027a led to upregulation of a nucleotide binding-leucine rich repeat domain protein coding gene (CaRLb1) and increased immunity against Phytophthora capsici in pepper leaves. Our study provided a highly sensitive and convenient method for sRNA research and identified new targets for genetic improvement of pepper immunity against P. capsici.

Development and Application of a Cleaved Amplified Polymorphic Sequence Marker (Phyto) Linked to the Pc5.1 Locus Conferring Resistance to Phytophthora capsici in Pepper (Capsicum annuum L.).

Phytophthora capsici causes destructive disease in several crop species, including pepper (Capsicum annuum L.). Resistance in this species is physiologically and genetically complex due to many P. capsici virulence phenotypes and different QTLs and R genes among the identified resistance sources. Several primer pairs were designed to follow an SNP (G/A) within the CA_011264 locus linked to the Pc5.1 locus. All primer pairs were designed on DNA sequences derived from CaDMR1, a homoserine kinase (HSK), which is a gene candidate responsible for the major QTL on chromosome P5 for resistance to P. capsici. A panel of 69 pepper genotypes from the Southern Seed germplasm collection was used to screen the primer pairs designed. Of these, two primers (Phyto_for_2 and Phyto_rev_2) surrounding the SNP proved successful in discriminating susceptible and resistant genotypes when combined with a restriction enzyme (BtgI). This new marker (called Phyto) worked as expected in all genotypes tested, proving to be an excellent candidate for marker-assisted selection in breeding programs aimed at introgressing the resistant locus into pure lines.

Compost Tea Induces Growth and Resistance against Rhizoctonia solani and Phytophthora capsici in Pepper

Compost teas (CTs) are organic solutions that constitute an interesting option for sustainable agriculture. Those that come from garden waste have been applied in vitro and in vivo on pepper plants to determine its suppressive effect against both Phytophthora capsici and Rhizoctonia solani. The studied CT showed relevant content in NO3−, K2O, humic acids, and microorganisms such as aerobic bacteria, N-fixing bacteria, and actinobacteria, which play a role in plant growth and resistance. This rich abundance of microbiota in the CT induced a reduction in the relative growth rate of both P. capsici and R. solani (31.7% and 38.0%, respectively) in in vitro assays compared to control. In addition, CT-irrigated plants displayed increased growth parameters and showed the first open flower one week before those treatments without CTs, which suggests that its application advanced the crop cycle. Concerning pathogen infection, damage caused by both pathogens became more apparent with a one-week inoculation compared to a four-week inoculation, which may indicate that a microbiological and chemical balance had been reached to cope with biotic stresses. Based on these results, we conclude that CT application induces plant growth and defense in pepper plants against P. capsici and R. solani because of its relevant soluble nutrient content and microbiota richness, which provides a novel point for plant nutrition and protection in horticultural crops.

Inhibitory effects of Bacillus licheniformis BL06 on Phytophthora capsici in pepper by multiple modes of action

Phytophthora capsici is a pathogenic oomycete that affects pepper cultivation and other commercial products, causing serious economic losses. Here, Bacillus licheniformis strain BL06 effectively reduced pepper Phytophthora blight severity. BL06 suppressed P. capsici mycelium growth by 70% in dual culture tests and induced excessive branching and significant lysis of P. capsici hyphae. The BL06 bacterial culture also inhibited in vitro sporangia development in a dose-dependent manner. Moreover, BL06 inhibited motility and displayed lytic activities against zoospores of P. capsici in a dose- and time-dependent manner. Indeed, the P. capsici infectiousness was reduced in host tissue treated by BL06, which is possibly associated with its ability to inhibit zoospore germination and germ tube growth in host tissues. Collectively, our results demonstrate the utility B. licheniformis BL06 as a potential biocontrol agent for the management of Phytophthora blight of peppers.

Identification of CBL and CIPK gene families and functional characterization of CaCIPK1 under Phytophthora capsici in pepper (Capsicum annuum L.)

BackgroundCalcineurin B-like proteins (CBLs) are major Ca2+ sensors that interact with CBL-interacting protein kinases (CIPKs) to regulate growth and development in plants. The CBL-CIPK network is involved in stress response, yet little is understood on how CBL-CIPK function in pepper (Capsicum annuum L.), a staple vegetable crop that is threatened by biotic and abiotic stressors.ResultsIn the present study, nine CaCBL and 26 CaCIPK genes were identified in pepper and the genes were named based on their chromosomal order. Phylogenetic and structural analysis revealed that CaCBL and CaCIPK genes clustered in four and five groups, respectively. Quantitative real-time PCR (qRT-PCR) assays showed that CaCBL and CaCIPK genes were constitutively expressed in different tissues, and their expression patterns were altered when the plant was exposed to Phytophthora capsici, salt and osmotic stress. CaCIPK1 expression changed in response to stress, including exposure to P. capsici, NaCl, mannitol, salicylic acid (SA), methyl jasmonate (MeJA), abscisic acid (ABA), ethylene (ETH), cold and heat stress. Knocking down CaCIPK1 expression increased the susceptibility of pepper to P. capsici, reduced root activity, and altered the expression of defense related genes. Transient overexpression of CaCIPK1 enhanced H2O2 accumulation, cell death, and expression of genes involved in defense.ConclusionsNine CaCBL and 26 CaCIPK genes were identified in the pepper genome, and the expression of most CaCBL and CaCIPK genes were altered when the plant was exposed to stress. In particular, we found that CaCIPK1 is mediates the pepper plant’s defense against P. capsici. These results provide the groundwork for further functional characterization of CaCBL and CaCIPK genes in pepper.

A Novel Transcription Factor CaSBP12 Gene Negatively Regulates the Defense Response against Phytophthora capsici in Pepper (Capsicum annuum L.).

SBP-box (Squamosa-promoter binding protein) genes are a type of plant-specific transcription factor and play important roles in plant growth, signal transduction and stress response. However, little is known about the SBP-box genes in pepper (CaSBP), especially in the process of Phytophthora capsici infection. In this study, a novel gene (CaSBP12) was selected from the CaSBP gene family, which was isolated from the pepper genome database in our previous study. The CaSBP12 gene was located in the nucleus of the cell and its silencing in the pepper plant enhanced the defense response against Phytophthora capsici infection. After inoculation with Phytophthora capsici, the root activity of the CaSBP12-silenced plants is compared to control plants, while malondialdehyde (MDA) content is compared viceversa. Additionally, the expression of defense related genes (CaPO1, CaSAR8.2, CaBPR1, and CaDEF1) in the silenced plants were induced to different degrees and the peak of CaSAR8.2 and CaBPR1 were higher than that of CaDEF1. The CaSBP12 over-expressed Nicotiana benthamiana plants were more susceptible to Phytophthora capsici infection with higher EC (electrical conductivity) and MDA contents as compared to the wild-type. The relative expression of defense related genes (NbDEF, NbNPR1, NbPR1a, and NbPR1b) in transgenic and wild-type Nicotiana benthamiana plants were induced, especially the NbPR1a and NbPR1b. In conclusion, these results indicate that CaSBP12 gene negative regulates the defense response against Phytophthora capsici infection which suggests their potentially significant role in plant defense. To our knowledge, this is the first report on CaSBP gene which negative regulate defense response.

Spermine and its interaction with proline induce resistance to the root rot pathogen Phytophthora capsici in pepper (Capsicum annuum)

The root rot pathogen Phytophthora capsici can severely damage production of peppers (Capsicum annuum). This study examined the effect of exogenous spermine and spermine in combination with proline on disease severity in pepper inoculated with P. capsici, as well as the effect of the treatments on P. capsici growth and zoospore development. Peroxidase, catalase, and polyphenol oxidase activity, the levels of phenolic compounds, malondialdehyde (MDA), and hydrogen peroxide (H2O2), and fresh and dry weight were measured on the 3rd, 5th, and 7th days after P. capsici infection. Disease severity and size of the necrotic lesions increased with the degree of injury caused by the pathogen and decreased in resistant varieties. The cultivar CM-334 exhibited the highest resistance to P. capsici followed by Amazon F1 whereas other genotypes were susceptible to the disease. Treatments with spermine and spermine + proline prior to inoculation decreased the severity of disease, the length of the necrotic lesions, and the MDA and H2O2 levels in all genotypes, and increased peroxidase, catalase, and polyphenol oxidase activity, the total level of phenolics, and fresh and dry weight, when compared to the untreated, inoculated plants. In addition, 1 mM spermine, 1 mM spermine + 1 mM proline, and 1 mM spermine + 10 mM proline decreased colony growth of P. capsici and zoospore production. This study showed that spermine and spermine + proline induced defense responses and increased resistance to P. capsici infection in pepper.

Phosphite-induced reactive oxygen species production and ethylene and ABA biosynthesis, mediate the control of Phytophthora capsici in pepper (Capsicum annuum).

Phytophthora capsici is an oomycete pathogen with a broad host range that inflicts significant damage in vegetables. Phosphite (Phi) is used to control oomycete diseases, but the molecular mechanisms underlying Phi-induced resistance to P. capsici are unknown. Thus, Phi-inhibited mycelial growth on strain LT1534 and primed host defence were analysed. We demonstrated that Phi (>5µgmL-1) had a direct antibiotic effect on mycelial growth and zoospore production, and that mortality and DNA content were significantly reduced by pre-treatment with Phi. In addition, elevated hydrogen peroxide (H2O2) promoted callose deposition and increased the levels of soluble proteins and Capsicum annuum L. pathogenesis-related 1 (CaPR1) expression. Furthermore, Phi (1gL-1) significantly increased the transcription of the antioxidant enzyme genes, and the genes involved in ethylene (ET) and abscisic acid (ABA) biosynthesis, as well as mitogen-activated protein kinase (MAPK) cascades. However, pre-treatment with reactive oxygen species (ROS), ABA and ET biosynthesis inhibitors decreased Phi-induced resistance and reduced the expression of ABA-responsive 1 (CaABR1) and lipoxygenase 1 (CaLOX1). In addition, the decreased ROS and ABA inhibited Phi-induced expression of CaMPK17-1. We propose that Phi-induced ROS production, ET and ABA biosynthesis mediate the control of P. capsici, and that ABA functions through CaMPK17-1-mediated MAPK signalling.

Combined use of bulked segregant analysis and microarrays reveals SNP markers pinpointing a major QTL for resistance to Phytophthora capsici in pepper.

Bulked segregant analysis (BSA) using Affymetrix GeneChips revealed candidate genes underlying the major QTL for Phytophthora capsici resistance in Capsicum. Using the candidate genes, reliable markers for Phytophthora resistance were developed and validated. Phytophthora capsici L. is one of the most destructive pathogens of pepper (Capsicum spp.). Resistance of pepper against P. capsici is controlled by quantitative trait loci (QTL), including a major QTL on chromosome 5 that is the predominant contributor to resistance. Here, to maximize the effect of this QTL and study its underlying genes, an F2 population and recombinant inbred lines were inoculated with P. capsici strain JHAI1-7 zoospores at a low concentration (3 × 10(3)/mL). Resistance phenotype segregation ratios for the populations fit a 3:1 and 1:1 (resistant:susceptible) segregation model, respectively, consistent with a single dominant gene model. Bulked segregant analysis (BSA) using Affymetrix GeneChips revealed a single position polymorphism (SPP) marker mapping to the major QTL. When this SPP marker (Phyto5SAR) together with other SNP markers located on chromosome 5 was used to confirm the position of the major QTL, Phyto5SAR showed the highest LOD value at the QTL. A scaffold sequence (scaffold194) containing Phyto5SAR was identified from the C. annuum genome database. The scaffold contained two putative NBS-LRR genes and one SAR 8.2A gene as candidates for contributing to P. capsici resistance. Markers linked to these genes were developed and validated by testing 100 F1 commercial cultivars. Among the markers, Phyto5NBS1 showed about 90% accuracy in predicting resistance phenotypes to a low-virulence P. capsici isolate. These results suggest that Phyto5NBS1 is a reliable marker for P. capsici resistance and can be used for identification of a gene(s) underlying the major QTL on chromosome 5.

Effectiveness of biological control of Phytophthora capsici in pepper by Trichoderma asperellum strain T34

Pepper ( Capsicum annuum L.), one of the most widely grown vegetables worldwide, is susceptible to root rot caused by Phytophthora capsici . Many biocides have recently been banned in Europe because of human health and environmental concerns. Integrated pest management is a European priority, where biological control together with other agronomic practices should replace pesticide management of plant diseases in the future. Application of different concentrations of the fungus Trichoderma asperellum strain T34 (the in T34 Biocontrol®) on incidence of disease caused by P. capsici in pepper was studied. Different methods of application of the microbial control agent and inoculation of the pathogen were examined. T34 and etridiazole (Terrazole®) were compared for their ability to suppress P. capsici . T34 reduced disease in most of the assayed situations (up to 71% disease reduction), while etridiazole was effective only when applied at the same time as the pathogen. The results obtained are discussed on the basis of the different modes of action of T34 and etridiazole. T34 is a useful biological alternative to chemicals for the control of P. capsici in pepper.

Screening potential bacterial biocontrol agents towards Phytophthora capsici in pepper

A total of 1,487 bacterial isolates were obtained from the rhizosphere, phyllosphere, endorhiza and endosphere of field-grown pepper. In a dual assay, 232 isolates displayed the antagonistic activity towards Phytophthora capsici L.; 36.6 % and 39.2 % of them were obtained from the rhizosphere and phyllosphere, respectively. 40 of the 232 antagonistic isolates producing inhibition zones of at least 5 mm in diameter were assessed for production of siderophores and chitinase, cellulose, and protease activity. These 40 isolates fell into 15 groups according to 90 % similarity of the banding patterns obtained by amplified ribosomal DNA restriction analysis (ARDRA). Seventeen isolates spanning the 15 groups were evaluated in greenhouse tests for their ability to control Phytophthora blight of pepper. Biocontrol efficacy ranged from 0.7 % to 92.3 %, with three isolates (B1301, R98, and PX35) exhibiting maximum ability to reduce the disease severity (83.5 %, 92.3 % and 83.5 %, respectively). Based on 16S rDNA sequencing, these isolates were identified as Bacillus cereus (B1301), Chryseobacterium sp (R98) and Bacillus cereus (PX35). This is the first report that Chryseobacterium sp. (R98) can function as a biocontrol agent of Phytophthora blight.

QTL Analysis for Resistance to Phytophthora capsici in Pepper Using a High Density SSR-based Map

A segregating doubled haploid (DH) population (n = 96) was developed by anther culture of an F1 plant crossed between susceptible (‘Manganji’) and resistant (‘Criollo de Morelos 334’) lines of pepper (Capsicum annuum L.) to conduct a genetic analysis of resistance to Phytophthora rot caused by Phytophthora capsici. In order to perform a quantitative trait locus (QTL) analysis, we constructed a high density simple sequence repeat (SSR)-based map with a total length of 878 cM. Sixteen linkage groups (LGs) and 118 SSR markers were located using the 626 SSR markers that we previously developed. Resistance was evaluated in two root inoculation tests. Interval mapping for the resistance to P. capsici detected a common major QTL in the duplicate tests and a minor QTL specific to the first test. The major QTL was located on LG15 and flanked with an SSR marker, CAMS420. In addition, seven SSR markers were located within 21 cM intervals from the peak of this QTL. In contrast, the QTL on LG3 was detected with small effects in the first test, the nearest marker was a dominant amplified fragment length polymorphism (AFLP) marker, and the QTL was surrounded by eight SSR markers within a distance of 10 cM. Since some of the linkage markers for agriculturally valuable traits cannot detect polymorphism within breeding populations in C. annuum, the present linkage markers may widen the choice in marker-assisted selection in breeding programs for Phytophthora rot resistant pepper cultivars.

Acibenzolar-S-methyl induced resistance to Phytophthora capsici in pepper leaves

The leaves of pepper (Capsicum anuum L.) were inoculated with Phytophthora capsici Leonian 3 d after treatment with acibenzolar-S-methylbenzo [1,2,3]thiadiazole-7-carbothioic acid-S-methyl ester (ASM) and resistance to Phytophthora blight disease was investigated. Results showed that P. capsici was significantly inhibited by ASM treatment by up to 45 % in planta. The pepper plants responded to ASM treatments by rapid and transient induction of L-phenylalanine ammonia-lyase (PAL), increase in total phenol content and activities of chitinase and β-1,3-glucanase. No significant increases in enzyme activities were observed in water-treated control plants compared with the ASM-treated plants. Therefore it may be suggested that ASM induces defense-related enzymes, PAL activity, PR proteins and phenol accumulation in ASM-treated plants and contribute to enhance resistance against P. capsici.

Isolation of Resistance Gene Analogs in Pepper Using Modified AFLPs

An efficient technique for isolation of resistant gene analogs (RGAs) in pepper from silver stained denaturing polyacrylamide gel was developed using a modified amplified fragment length polymorphism (AFLP) strategy. Pepper DNA was digested, ligated and pre-amplified as in a normal AFLP method. The selective amplification was made by using combinations with oligonucleotide primers based on conserved motifs in and around nucleotide binding site (NBS) of known NBS-leucine-rich repeats resistance proteins from known resistant genes. The amplified products were separated by using denaturing polyacrylamide gels and silver staining instead of radioactive labelling. We isolated specific polymorphic AFLP bands directly from the gels with one round of polymerase chain reaction amplification, in order to confirm, after sequencing, that these bands have homologies with products of resistance genes described so far. Two bands (R2: 250 bp and R6: 150 bp) are particularly highlighted because they could be considered as RGAs related to resistance to Phytophthora capsici in pepper, because their sequences have a very high homology with other resistant gene analogs that have already been described. Besides, they were only detected in the resistant parent and in the bulked resistant segregants but not in the susceptible parent or susceptible F2 segregants. We can conclude that the technique used is clean, quick and efficient for the isolation of RGAs in pepper.

Ecologically Based Approaches to Management of Phytophthora Blight on Bell Pepper.

HomePlant DiseaseVol. 83, No. 12Ecologically Based Approaches to Management of Phytophthora Blight on Bell Pepper PreviousNext OPENOpen Access licenseEcologically Based Approaches to Management of Phytophthora Blight on Bell PepperJean Beagle Ristaino and Stephen A. JohnstonJean Beagle RistainoSearch for more papers by this author and Stephen A. JohnstonSearch for more papers by this authorAffiliationsAuthors and Affiliations Jean Beagle Ristaino , North Carolina State University, Raleigh Stephen A. Johnston , Rutgers Agricultural Research and Extension Center, Bridgeton, NJ Published Online:23 Feb 2007https://doi.org/10.1094/PDIS.1999.83.12.1080AboutSectionsPDF ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat DetailsFiguresLiterature CitedRelated Vol. 83, No. 12 December 1999SubscribeISSN:0191-2917e-ISSN:1943-7692 Metrics Article History Issue Date: 25 Jan 2008Published: 23 Feb 2007 Pages: 1080-1089 Information© 1999 The American Phytopathological SocietyPDF downloadCited byMajor Soilborne Pathogens of Field Processing Tomatoes and Management Strategies19 January 2023 | Microorganisms, Vol. 11, No. 2New metabolites from Streptomyces pseudovenezuelaeNA07424 and their potential activity of inducing resistance in plants against Phytophthora capsici13 October 2022 | Pest Management Science, Vol. 79, No. 1Sympatric occurrence of sibling Phytophthora species associated with foot rot disease of black pepper in India23 February 2022 | Brazilian Journal of Microbiology, Vol. 53, No. 2Phytophthora capsici on Capsicum Plants: A Destructive Pathogen in Chili and Pepper Crops5 May 2022Identification of molecular marker linked to resistance gene loci against Indian isolate of Phytophthora capsici L. causing root rot in chilli (Capsicum annuum L.)13 November 2021 | Australasian Plant Pathology, Vol. 51, No. 2Broad Resistance to Post-Harvest Fruit Rot in USVL Watermelon Germplasm Lines to Isolates of Phytophthora capsici Across the United StatesChandrasekar S. 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