Diversity and activity of biosurfactant-producing Pseudomonas in the rhizosphere of black pepper in Vietnam

Aims: Phytophthora capsici is a major pathogen of black pepper and zoospores play an important role in the infection process. Fluorescent pseudomonads that produce biosurfactants with zoosporicidal activities were isolated from the black pepper rhizosphere in Vietnam, and their genotypic diversity and potential to control Phy. capsici root rot was determined. Methods: Biosurfactant‐producing pseudomonads were genotypically and biochemically characterized by BOX‐polymerase chain reaction (PCR), 16S‐rDNA sequencing, reverse‐phase‐high‐performance liquid chromatography and liquid chromatography‐masss spectrometry analyses. Results: Biosurfactant‐producing fluorescent pseudomonads make up c. 1.3% of the culturable Pseudomonas population in the rhizosphere of black pepper. Although BOX‐PCR revealed substantial genotypic diversity, the isolates were shown to produce the same biosurfactants and were all identified as Pseudomonas putida. When applied to black pepper stem cuttings, several of the biosurfactant‐producing strains provided significant disease control. In absence of the disease, several of the bacterial strains promoted shoot and root growth of black pepper stem cuttings. Conclusions: Biosurfactant‐producing pseudomonads indigenous to the rhizosphere of black pepper plants are genotypically diverse and provide a novel resource for the control of Phy. capsici root rot and growth promotion of black pepper stem cuttings. Significance and Impact of the Study: The results of this study provide a strong basis for further development of supplementary strategies with antagonistic bacteria to control foot and root rot of black pepper and to promote plant growth.     

Compatibility of <Emphasis Type="Italic">Piriformospora indica</Emphasis> and <Emphasis Type="Italic">Trichoderma harzianum</Emphasis> as dual inoculants in black pepper (<Emphasis Type="Italic">Piper nigrum</Emphasis> L.)

The compatibility of two biological inoculants, Trichoderma harzianum, a mycoparasitic biological control fungus and Piriformospora indica, a root colonizing plant-growth promoting endophytic fungus was evaluated using tissue cultured black pepper plantlets. We report, for the first time, the ability of P. indica to colonize black pepper, a perennial climber. T. harzianum inhibited the growth of P. indica in an in vitro dual culture plate assay. Simultaneous inoculation with both biological inoculants of tissue cultured black pepper plantlets negatively influenced root colonization by P. indica. However, when P. indica was applied initially followed 30 days later by T. harzianum, there was increased root colonization by the root endophyte P. indica and beneficial effects were found on the growth of the black pepper plants. The present study also showed that the efficacy of inoculation of the two fungal biological agents can be increased by sequential application of P. indica at the hardening stage followed by T. harzianum during transplanting into a soil-sand mixture.     

Endophytic bacterial flora in root and stem tissues of black pepper (Piper nigrum L.) genotype: isolation, identification and evaluation against Phytophthora capsici

Aim:  To isolate and identify black pepper ( Piper nigrum L) associated endophytic bacteria antagonistic to Phytophthora capsici causing foot rot disease.
Methods and Results:  Endophytic bacteria (74) were isolated, characterized and evaluated against P. capsici . Six genera belong to Pseudomonas spp (20 strains), Serratia (1 strain), Bacillus spp. (22 strains), Arthrobacter spp. (15 strains), Micrococcus spp. (7 strains), Curtobacterium sp. (1 strain) and eight unidentified strains were isolated from internal tissues of root and stem. Three isolates, IISRBP 35, IISRBP 25 and IISRBP 17 were found effective for Phytophthora suppression in multilevel screening assays which recorded over 70% disease suppression in green house trials. A species closest match (99% similarity) of IISRBP 35 was established as Pseudomonas aeruginosa ( Pseudomonas EF568931), IISRBP 25 as P. putida ( Pseudomonas EF568932), and IISRBP 17 as Bacillus megaterium ( B. megaterium EU071712) based on 16S rDNA sequencing.
Conclusion:  Black pepper associated P. aeruginosa , P. putida and B. megaterium were identified as effective antagonistic endophytes for biological control of Phytophthora foot rot in black pepper.
Significance and Impact of the Study:  This work provides the first evidence for endophytic bacterial diversity in black pepper stem and roots, with biocontrol potential against P. capsici infection.     

Greenhouse and field evaluations of potassium phosphonate: the control of Phytophthora foot rot of black pepper in Vietnam

Phytophthora foot rot of black pepper caused by Phytophthora capsici is a major disease of black pepper throughout production areas in Vietnam. The disease causes collar, foot and tap root rots and eventual death of the infected vine. Potassium phosphonate was evaluated for the control of this disease in greenhouse and field trials. In greenhouse trials three-month-old vines treated with phosphonate by soil drenching (10–20 g a.i./l) and then inoculated with P. capsici mycelium (2% v/v soil) had significantly less foot rot compared to vines grown in non-treated soil. In field trials mature vines were treated with phosphonate at 50–100 g a.i/pole soil drenching or 10 g a.i./l by root infusion. After 10 days root, stem and leaf specimens were removed for bioassay by inoculation with 5 ml of P. capsici zoospores suspension (10⁶–10⁸ spores/ml). Soil drenching with phosphonate inhibited the colonisation of pathogen on excised leaf, stem and root tissues, significantly more than phosphonate root infusion. Our study provides further evidence supporting the efficacy of potassium phosphonate in the management of black pepper foot rot caused by P. capsici. The excised leaf and stem bioassay used in this study is a rapid and useful technique for testing the efficacy of systemic fungicides in controlling this disease.     

Endophytic bacteria of <Emphasis Type="Italic">Mammillaria fraileana</Emphasis>, an endemic rock-colonizing cactus of the southern Sonoran Desert

The small cactus Mammillaria fraileana is a pioneer rock-colonizing plant harboring endophytic bacteria with the potential for nitrogen fixation and rock weathering (phosphate solubilization and rock degradation). In seeds, only a combination of culture-independent methods, such as fluorescence in situ hybridization, scanning electron microscopy, and fluorescence vital staining, detected significant amounts of non-culturable, but living, endophytic bacteria distributed underneath the membrane covering the embryo, in the undifferentiated tissue of the embryo, and in the vascular tissue. Large populations of culturable endophytic bacteria were detected in stems and roots of wild plants colonizing rocks in the southern Sonoran Desert, but not in seeds. Among 14 endophytic bacterial isolates found in roots, four isolates were identified by full sequencing of their 16S rRNA gene. In vitro tests indicated that Azotobacter vinelandii M2Per is a potent nitrogen fixer. Solubilization of inorganic phosphate was exhibited by Pseudomonas putida M5TSA, Enterobacter sakazakii M2PFe, and Bacillus megaterium M1PCa, while A. vinelandii M2Per, P. putida M5TSA, and B. megaterium M1PCa weathered rock by reducing the size of rock particles, probably by changing the pH of the liquid media. Cultivated seedlings of M. fraileana, derived from disinfected seeds and inoculated with endophytic bacteria, showed re-colonization 105 days after inoculation. Their densities decreased from the root toward the stem and apical zones. Functional traits in planta of culturable and non-culturable endophytic bacteria in seeds remain unknown.     

Control of Late Blight (Phytophthora capsici ) in Pepper Plant with a Compost Containing Multitude of Chitinase-producing Bacteria

Compost sustaining a multitude of chitinase-producing bacteria was evaluated in a greenhouse study as a soil amendment for the control of late blight (Phytophthora capsici L.) in pepper (Capsicum annuum L.). Microbial population and exogenous enzyme activity were measured in the rhizosphere and correlated to the growth and health of pepper plant. Rice straw was composted with and without a chitin source, after having been inoculated with an aliquot of coastal area soil containing a known titer of chitinase-producing bacteria. P. capsici inoculated plants cultivated in chitin compost-amended soil exhibited significantly higher root and shoot weights and lower root mortality than plants grown in pathogen-inoculated control compost. Chitinase and β-1,3-glucanase activities in rhizosphere of plants grown in chitin compost-amended soil were twice that seen in soil amended with control compost. Colony forming units of chitinase-producing bacteria isolated from rhizosphere of plants grown in chitin compost-amended soil were 10³ times as prevalent as bacteria in control compost. These results indicate that increasing the population of chitinase-producing bacteria and soil enzyme activities in rhizosphere by compost amendment could alleviate pathogenic effects of P. capsici.     

Treatment of Paenibacillus illinoisensis suppresses the activities of antioxidative enzymes in pepper roots caused by Phytophthora capsici infection

Summary The activity of antioxidative enzymes after inoculation of pepper (Capsicum annuum L. Chungok) with a pathogen, Phytophthora capsici (P), the causal agent of Phytophtora blight and dual inoculation of pathogen and an antagonist, Paenibacillus illinoisensis KJA-424 (P+A), were measured and compared with that of non-inoculated (C) roots. Root mortality was significantly reduced by about 84% in P+A treatment compared with P treatment alone. When compared to the non-inoculated (C) roots, malondialdehyde (MDA) concentration gradually decreased by 52.4% in 7 days only in P-treated roots and hydrogen peroxide (H₂O₂) was not significantly affected by the treatment for 5 days but significantly decreased in the P+A-treated roots at day 7. P-treatment continuously induced peroxidase (POD) and superoxide dismutase (SOD), resulting in significant increases of 36.7% and 27.7% at day 7, respectively, compared to the control. In P+A-treated roots, the activities of POD and SOD also increased for 5 days but returned to the control level at day 7. Catalase activity fluctuated but again increased over the 7-day period following P+A inoculation. These results indicate that an antagonist P. illinoisensis KJA-424 alleviated root mortality and suppressed the elevated activities of POD and SOD in the root of pepper plant root caused by P.␣capsici infection.     

Biocontrol of Late Blight Disease (Phytophthora capsici) of Pepper and the Plant Growth Promotion by Paenibacillus ehimensis KWN38

For field application of a bacterial strain used to control Phythophthora capsici, we will need a biologically and economically efficient carrier medium. The known antagonist Paenibacillus ehimensisKWN38 was grown in a grass medium where it showed high antifungal and lytic enzyme activities. To demonstrate the potential of P. ehimensisKWN38 for biocontrol of late blight disease in pepper, pot trials were conducted by treating the 1‐month‐old plants with water (W), a selected grass medium (G3), G plus P. ehimensisKWN38 inoculation (G3P) or synthetic fungicide (F). The shoot dry weight in G3P was higher than that in W and F treatments at 15 days after zoospore infection (DZI). The root dry weight in G3P was also higher than that in W. The root mortality of G3 and W increased over 58 and 80% at 15 DZI, and some plants in those treatments wilted due to the failure of root physiology. The plants in G3P and F survived well because of their better root health conditions. Soil cellulase activity of G3P was consistently higher than that of W and F at earlier observation times (0, 2 and 6 DZI). The root β‐1,3‐glucanase activity of G3P promptly increased to maximum shortly after zoospore infection and reached the maximum value of 51.12 unit g^?1 of fresh weight at 2 DZI. All these results indicate that inoculation of P. ehimensisKWN38 to the root zone of potted pepper plants increases plant growth, root and soil enzyme activities and alleviates the root death caused by infection with P. capsici zoospores.     

Plant growth promoting rhizhobacteria (PGPR)-mediated root proliferation in black pepper (Piper nigrum L.) as evidenced through GS Root software

Abstract

Pseudomonas fluorescens strains which are proven biocontrol agents in black pepper against foot rot (caused by Phytophthora capsici ) were also found to enhance root proliferation and fibre root production. Experiments conducted in the greenhouse with five efficient strains of P. fluorescens (IISR-6, IISR-8, IISR-11, IISR-13 and IISR-51) showed that the bacterial strains could significantly increase the root biomass of the plants (30 – 135%). Parameters for total root length, root area and root tips were estimated by scanning the entire root system and analysis through GS Root® software (PP systems, Winterstreet, USA). All the strains increased the root length in the treated plants (12 – 127%), the highest being with IISR-6, which was on a par with IISR-11 and IISR-51. A similar trend was observed with the total root area after bacterization (43 – 200%). The P. fluorescens treated plants had a higher number of feeder roots as evidenced by the increased number of root tips (82 – 137%). The enhanced growth parameters upon root bacterization could be corroborated with the production of the plant growth hormones IAA & GA by the bacterial strains and their P-solubilization potential.     

Broad-spectrum antimicrobial activity of volatile organic compounds from endophytic Pseudomonas putida BP25 against diverse plant pathogens

ABSTRACT

Black pepper endophytic Pseudomonas putida BP25 produced diverse antimicrobial volatile organic compounds having potential for plant disease management. Chemically synthesised volatiles such as 2, 5-dimethyl pyrazine; 2-methyl pyrazine; dimethyl trisulphide; 2-ethyl 5-methyl pyrazine; and 2-ethyl 3, 6-dimethyl pyrazine showed inhibitory activity against oomycete pathogens, Phytophthora capsici & Pythium myriotylum; fungal pathogens, Rhizoctonia solani, Colletotrichum gloeosporioides, Athelia rolfsii, Gibberella moniliformis & Magnaporthe oryzae; bacterial pathogen, Ralstonia pseudosolanacearum and plant parasitic nematode, Radopholus similis. Among them, dimethyl trisulphide completely inhibited oomycete and fungal pathogens as well as R. similis at a concentration of 2.65?µg?cm^?3 under in vitro conditions. Pyrazines suppressed Phytophthora lesions on shoot cuttings of black pepper upon in planta volatile treatment. Dimethyl trisulphide was the only compound that exhibited soil fumigant activity against P. capsici, R. solani and A. rolfsii (6.25?µg?cm^?3), C. gloeosporioides and G. moniliformis (12.5?µg?cm^?3), and R. similis (50?µg?cm^?3). Altogether, endophytic Pseudomonas putida BP25 and its volatile organic compounds offer an alternative strategy for eco-friendly disease management in agriculture.     

The volatile-producing Flavobacterium johnsoniae strain GSE09 shows biocontrol activity against Phytophthora capsici in pepper

Aims: Previously, we selected a bacterial strain (GSE09) antagonistic to Phytophthora capsici on pepper, which produced a volatile compound (2,4‐di‐tert‐butylphenol), inhibiting the pathogen. In this study, we identified strain GSE09 and characterized some of the biological traits of this strain in relation to its antagonistic properties against P. capsici. In addition, we examined bacterial colonization on the root surface or in rhizosphere soil and the effect of various concentrations of the volatile compound and strain GSE09 on pathogen development and radicle infection as well as radicle growth. Methods and Results: Strain GSE09 was identified as Flavobacterium johnsoniae, which forms biofilms and produces indolic compounds and biosurfactant but not hydrogen cyanide (HCN) with little or low levels of antifungal activity and swimming and swarming activities. Fl. johnsoniae GSE09 effectively colonized on pepper root, rhizosphere, and bulk (pot) soil, which reduced the pathogen colonization in the roots and disease severity in the plants. Various concentrations of 2,4‐di‐tert‐butylphenol or strain GSE09 inhibited pathogen development (mycelial growth, sporulation, and zoospore germination) in I‐plate (a plastic plate containing a center partition). In addition, germinated seeds treated with the compound (1–100 μg ml^?1) or the strain (10²–10¹⁰ cells ml^?1) significantly reduced radicle infection by P. capsici without radicle growth inhibition. Conclusions: These results indicate that colonization of pepper root and rhizosphere by the Fl. johnsoniae strain GSE09, which can form biofilms and produce indolic compounds, biosurfactant, and 2,4‐di‐tert‐butylphenol, might provide effective biocontrol activity against P. capsici. Significance and Impact of the Study: To our knowledge, this is the first study demonstrating that the Fl. johnsoniae strain GSE09, as a potential biocontrol agent, can effectively protect pepper plants against P. capsici infection by colonizing the roots.     

Control of leaf-tip necrosis of micropropagated ornamental statice by elimination of endophytic bacteria

Summary Leaf-tip necrosis of micropropagated statice plantlets is a serious problem in commercial laboratories in Taiwan. Endophytic bacteria were detected in plantlets obtained from commercial laboratories with a leaf-tip necrosis problem. Endophytic bacteria were detected in flower stalks collected from four different statice farms at frequencies ranging from 61 to 100%. All plantlets regenerated from flower-stalk explants that tested free of endophytic bacteria did not develop leaf-tip necrosis. The most frequently detected endophytic bacteria were Pasteurella multocida, Stenotrophomonas maltophilia, and Alcaligenes sp. Most endophytic bacteria in statice plantlets were eliminated by the subculture of plantlets on medium with augmentin, cefotaxime, or augmentin plus cefotaxime. Those plantlets freed from endophytic bacteria by subculture on antibiotic-amended medium did not develop leaf-tip necrosis. Our results show that leaf-tip necrosis of micropropagated statice plantlets is associated with endophytic bacteria, and that the disease can be controlled by using explants pre-tested to be free from endophytic bacteria or by the subculture of affected plantlets on antibiotic-amended medium.     

火棘不同组织内生细菌群落多样性

为了解火棘不同组织内生细菌群落多样性,该研究采用高通量测序技术对火棘内生细菌16S rRNA V5～V7可变区进行测序,分析火棘不同组织部位内生细菌群落多样性。结果表明:(1)从火棘根、茎、叶组织中共获得内生细菌OTU 1 818个,其中根部754个,茎部 308 个,叶部756个,三者共有 OTU 152 个。(2)物种分类显示,不同火棘组织内生细菌具有丰富的群落多样性,火棘根部内生细菌种类隶属于23门53纲137目216科373属557种,其中异样根瘤菌属(Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium)和链霉菌属(Streptomyces)为优势属,其相对丰度分别为 10.57%和 8.00%; 茎部内生细菌种类隶属于21门32纲76目126科204属270种,其中马赛菌属(Massilia)和未知分类的丛毛单胞菌科属(unclassified_f_Comamonadaceae)为优势属,其相对丰度分别为31.10%和12.82%; 叶部内生细菌种类隶属于21门52纲130目210科380属581种,其中土芽孢杆菌属(Geobacillus)和假单胞菌属(Pseudomonas)为优势属,其相对丰度分别为12.31%和9.84%。(3)PICRUSt功能预测表明,根部内生细菌物种最丰富,参与各种代谢调控的细菌丰度最高。该研究结果为进一步探讨植物内生细菌功能,挖掘新的有益微生物资源提供了参考。     

Selecting Bacterial Strains for Use in the Biocontrol of Diseases Caused by <Emphasis Type="Italic">Phytophthora capsici</Emphasis> and <Emphasis Type="Italic">Alternaria alternata</Emphasis> in Sweet Pepper Plants

More than 500 isolates of bacteria were obtained from the aerial part and rhizosphere of sweet pepper (Capsicum annuum L.) plants harvested from different places in the Region of Murcia (Spain). The isolates were purified and assayed in vitro against Phytophthora capsici and Alternaria alternata. Sixty isolates (12 %) produced an inhibition zone against at least one of the pathogens, while ten had a strongly inhibitory effect on both pathogens assayed. Microscopic observation of interactions zone showed cell vacuolisation, hyphae lysis and spilling of cytoplasm content of the pathogens in the culture media. These ten isolates were then chosen for biocontrol of Phytophthora root rot and Alternaria leaf spots of pepper plants in vivo. Four of them denominated HS93, LS234, LS523 and LS674 reduced P. capsici root rot by 80, 51, 49 and 54 %, respectively, and A. alternata leaf spots by 54, 74, 62 and 53 %. HS93 belongs to the genus Bacillus and probably the species subtilis, while LS234, LS523 and LS674 belong to the genus Bacillus and probably the species licheniformis. Dry mass of plants treated with these bacteria was significantly higher than that of non-treated and inoculated plants.     

Optimization of Culturing Conditions of a Strain of Phytophthora capsici Pathogenic to Habanero Pepper (Capsicum chinense)

Phytophthora capsici is an oomycete known as the causal agent of wilting disease in Capsicum spp., which causes rotting of roots, crowns, stems, leaves and fruits. To date, little is known about the production of phytotoxic metabolites by P. capsici or their role in the infection process. As part of a project directed towards the isolation and identification of phytotoxins produced by a strain of P. capsici pathogenic to habanero pepper (Capsicum chinense), we have evaluated the effect of factors such as aeration, light and culture medium on the production of mycelium and phytotoxic metabolites by P. capsici. The results showed that culturing P. capsici in potato dextrose broth (PDB) containing habanero pepper leaf infusion, in the dark and under still conditions, results in a high production of mycelium and a high phytotoxicity of the culture filtrate, in the shortest period of time.     

Identification of IAA-producing endophytic bacteria from micropropagated Echinacea plants using 16S rRNA sequencing

The presence of latent bacteria is a serious problem in plant tissue cultures. While endophytes are generally beneficial to plants in situ, they may affect culture growth under the modified conditions in vitro. The present study was undertaken to identify and characterize endophytic bacteria associated with the medicinal plant Echinacea in tissue culture. Based on classical microbiological tests and 16S rRNA analyses, it was found that endophytic bacteria associated with aseptically micropropagated Echinacea plantlets are representatives of several genera, Acinetobacter, Bacillus, Pseudomonas, Wautersia (Ralstonia) and Stenotrophomonas. Based on TLC and HPLC analyses, we found that Pseudomonas stutzeri P3 strain produces plant hormone, auxin (indole-3-acetic acid, IAA). Antibiotic resistance was also assessed as a virulence factor. The majority of endophytic bacteria were resistant to the antibiotic kanamycin, but susceptible to chloramphenicol. Recommendations for propagating Echinacea in vitro cultures involve the addition of chloramphenicol, tetracycline, and ampicillin, antibiotics that cause no side effects on these plant species.     

Biological Control of Phytophthora Root Rot of Pepper Using Trichoderma harzianum and Streptomyces rochei in Combination

A combination of two compatible micro‐organisms, Trichoderma harzianum and Streptomyces rochei, both antagonistic to the pathogen Phytophthora capsici, was used to control root rot in pepper. The population of the pathogen in soil was reduced by 75% as a result. Vegetative growth of the mycelium of P. capsici was inhibited in vitro on the second day after P. capsici and T. harzianum were placed on the opposite sides of the same Petri plate. Trichoderma harzianum was capable of not only arresting the spread of the pathogen from a distance, but also after invading the whole surface of the pathogen colony, sporulating over it. Scanning electron microscopy showed the hyphae of P. capsici surrounded by those of T. harzianum, their subsequent disintegration, and the eventual suppression of the pathogen's growth. Streptomyces rochei produced a zone of inhibition, from which was obtained a compound with antioomycete property secreted by the bacteria. When purified by high‐pressure liquid chromatography, this compound was identified as 1‐propanone, 1‐(4‐chlorophenyl), which seems to be one of the principal compounds involved in the antagonism. A formulation was prepared that maintained the compound's capacity to inhibit growth of the pathogen for up to 2 years when stored at room temperature in the laboratory on a mixture of plantation soil and vermiculite. The two antagonists, added as a compound formulation, were effective at pH from 3.5 to 5.6 at 23–30°C. The optimal dose of the antagonists in the compound formulation was 3.5 × 10⁸ spores/ml of T. harzianum and 1.0 × 10⁹ FCU/ml of S. rochei. This is the first report of a compound biocontrol formulation of these two antagonists with a potential to control root rot caused by P. capsici.     

QTL mapping of anthracnose (<Emphasis Type="Italic">Colletotrichum</Emphasis> spp.) resistance in a cross between <Emphasis Type="Italic">Capsicum annuum</Emphasis> and <Emphasis Type="Italic">C. chinense</Emphasis>

Anthracnose fruit rot is an economically important disease that affects pepper production in Indonesia. Strong resistance to two causal pathogens, Colletotrichum gloeosporioides and C. capsici, was found in an accession of Capsicum chinense. The inheritance of this resistance was studied in an F₂ population derived from a cross of this accession with an Indonesian hot pepper variety (Capsicum annuum) using a quantitative trait locus (QTL) mapping approach. In laboratory tests where ripe fruits were artificially inoculated with either C. gloeosporioides or C. capsici, three resistance-related traits were scored: the infection frequency, the true lesion diameter (averaged over all lesions that actually developed), and the overall lesion diameter (averaged over all inoculation points, including those that did not develop lesions). One main QTL was identified with highly significant and large effects on all three traits after inoculation with C. gloeosporioides and on true lesion diameter after inoculation with C. capsici. Three other QTL with smaller effects were found for overall lesion diameter and true lesion diameter after inoculation with C. gloeosporioides, two of which also had an effect on infection frequency. Interestingly, the resistant parent carried a susceptible allele for a QTL for all three traits that was closely linked to the main QTL. The results with C. capsici were based on less observations and therefore less informative. Although the main QTL was shown to have an effect on true lesion diameter after inoculation with C. capsici, no significant QTL were identified for overall lesion diameter or infection frequency.     

Biocontrol activity and root colonization by Pseudomonas corrugata strains CCR04 and CCR80 against Phytophthora blight of pepper

Previously, we selected Pseudomonas corrugata strains CCR04 and CCR80 as rhizobacteria suppressive to Phytophthora blight of pepper caused by Phytophthora capsici. In this study, we investigated soil microbial activity in pepper plants root-drenched with strains CCR04 and CCR80 in relation to their biocontrol activity, root colonization by using bacterial population counts and scanning electron microscopy, biofilm formation and cell motility as well as cell sensitivity to hydrogen peroxide (H₂O₂). As a result, strains CCR04 and CCR80 more effectively suppressed disease expression in pepper plants through root colonization than did Paenibacillus polymyxa AC-1 (positive control), Escherichia coli DH5α (negative control) or MgSO₄ solution (untreated control). Strains CCR04 and CCR80 had efficient biofilm formation and cell motility (swimming and swarming activities) abilities and responded to certain tested compounds (amino acids, organic acids and sugars), which can be found in root exudates. Strains CCR04 and CCR80 and the positive control strain AC-1 were relatively insensitive to H₂O₂, a reactive oxidative species at concentration up to 20 mM, unlike the negative control strain DH5α. Taken together, these results suggest that P. corrugata CCR04 and CCR80 can effectively inhibit P. capsici infection of pepper plants through successful colonization of plant roots. This bacterial colonization may be facilitated by the biofilm formation ability and cell motility in addition to reduced sensitivity to H₂O₂ and probably the production of antimicrobial compounds. These findings highlight the potential of strains CCR04 and CCR80 as biocontrol agents for the management of Phytophthora blight of pepper.     

Inoculation of Paenibacillus illinoisensis alleviates root mortality, activates of lignification-related enzymes, and induction of the isozymes in pepper plants infected by Phytophthora capsici

To investigate the variations of the enzymes responsible for lignification, after inoculation with Phytophthora capsici and/or Paenibacillus illinoisensis KJA-424, in relation to biocontrol of Phytophthora blight in pepper, roots of two-month-old plants were inoculated with P. capsici inoculation (P), and co-inoculation of P. capsici and P. illinoisensis cell cultures (P + A). Root mortality of pepper plants induced by inoculation of P. capsici was completely recovered by co-inoculation with antagonistic KJA-424. At day 7, peroxidase (POD) activity increased by 36.7% in P-treated roots but by 7.1% only in P + A-treated, compared with control. Polyphenol oxidase (PPO) activity increased for 3 days and then drastically decreased in P-treated roots but maintained a constant level in control and P + A-treated. At day 7, PPO activity in P-treated leaves decreased but recovered to the level of control in the P + A-treated. Three major POD isozymes (45, 53, and 114 kDa) were shown in P-treated roots, while two major (53 and 114 kDa) in control and P + A-treated, suggesting that the 45 kDa of POD was actively induced in P-treated roots but not induced in P + A-treated roots. A PPO isozyme of 80 kDa was induced in P-treated roots but not induced by co-treated with KJA-424. In leaves, the POD isozyme of 45 kDa appears to be systemically induced in P-treated only. The PPO isozyme of 80 kDa in leaves was not induced by pathogen challenge but recovered by co-inoculated with P. illinoisensis. All these results suggest that the inoculation of an antagonist, P. illinoisensis alleviates root mortality, activates of lignification-related enzymes and induction of the isozymes in pepper plants infected by P. capsici.