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1.
Effects simultaneous and sequential inoculations of Meloidogyne incognita, Ralstonia solanacearum and Phomopsis vexans were studied on the growth, chlorophyll and carotenoid contents of eggplants grown in 25% fly ash and 25% sand mix soil. Plants grown in 25% fly ash mix soil had lesser plant growth than grown in 25% sand ash mix soil. Inoculation of M. incognita / R. solanacearum or P. vexans caused reduction in plant growth, chlorophyll and carotenoid contents in both types of soils but these pathogens in combination caused a greater reduction in than individual inoculation. Inoculation of M. incognita 20 days prior to R. solanacearum caused a greater reduction in plant growth than inoculation of M. incognita prior to P. vexans. Inoculation of P. vexans prior to R. solanacearum caused a lesser reduction in plant growth, chlorophyll and carotenoid contents than inoculation of P. vexans prior to M. incognita. Inoculation of R. solanacearum 20 days prior to M. incognita caused a greater reduction in plant growth, chlorophyll and carotenoid contents than inoculation of R. solanacearum prior to P. vexans. Galling and multiplication of M. incognita was higher in plants grown in 25% sand amended soil than with 25% fly ash soil. R. solanacearum and P. vexans had adverse effects on galling and nematode multiplication. Wilt and blight indices caused by R. solanacearum and P. vexans were 3 respectively. Wilt and blight indices were 4 when two pathogens were inoculated together.  相似文献   

2.
Ralstonia solanacearum, the devastating causal agent of potato bacterial wilt, is a soil-borne bacterium that can survive in the soil for a long time. The development of sensitive on-field detection methods for this pathogen is highly desirable due to its widespread host range and distribution. A novel nanobiosensor was thus developed to detect unamplified genomic DNA of R. solanacearum in farm soil. Gold nanoparticles functionalized with single-stranded oligonucleotides served as a probe to detect R. solanacearum genomic DNA. The advantages of this strategy include rapidity, facile usage and being a visual colorimetric method.  相似文献   

3.
Bacillus amyloliquefaciens IUMC7 and its culture supernatant, which we previously found to suppress Fusarium wilt, were found here to have antimicrobial activity against Ralstonia solanacearum. In in vivo experiments, mushroom compost inoculated with IUMC7 significantly reduced disease severity in tomato plants over that in control soils. The R. solanacearum population decreased in soil inoculated with IUMC7. A TLC–bioautography assay showed that one of the antimicrobial substances produced by IUMC7 is likely an iturin-like lipopeptide. These results suggest that these antimicrobial compounds are responsible for disease suppression and that mushroom compost containing IUMC7 has potential as a biocontrol product.  相似文献   

4.
Ralstonia solanacearum “species complex” (RSSC) represents soil-borne plant pathogenic bacteria, consisting of diverse and widespread strains that cause bacterial wilt on a wide range of host plants. A recent polyphasic taxonomic study has divided the RSSC into three bacterial species; Ralstonia pseudosolanacearum (phylotypes I and III), Ralstonia solanacearum (phylotype II) and Ralstonia syzygii (phylotype IV). Currently, standard identification of RSSC in plant health laboratories mainly relies on performance of two tests that are based on a different principle. However, these tests are inadequate to precisely discriminate among the three bacterial species in the RSSC. The accurate identification of each of the three bacterial species in the RSSC requires additional molecular tests, including a phylotype determination. These methodologies are labor-intensive, time consuming and rather impractical for routine identification purposes in a plant health laboratory. We explored the potential for an accurate identification of R. pseudosolanacearum (phylotypes I and III) and R. solanacearum (phylotype II) in RSSC, upon implementation of the MALDI-TOF MS tool, and after the creation and validation of an in-house database supplementing the commercial database and covering the entire known genetic diversity in RSSC. MALDI-TOF MS is an emerging approach for identification of bacterial plant pathogens and has been shown to be robust and reproducible. Additionally, when compared to the conventional microbial identification methods it is shown to be less laborious and less expensive. Validation data demonstrated that our in-house database (Mass Spectra Profiles, MSPs) was very specific resulting in the rapid and accurate identification of Ralstonia solanacearum (phylotype II), and Ralstonia pseudosolanacearum (phylotypes I and III). Additionally, no false positive results were obtained with our in-house database for other related Ralstonia sp., such as the R. picketii isolate PD 3286, or for the Pseudomonas syringae and Pseudomonas spp. isolates.  相似文献   

5.
Bacterial wilt, caused by Ralstonia solanacearum, is a devastating disease resulting in tremendous losses of economic crops such as plants in the Solanaceae. Recent studies showed that R. solanacearum is spreading from the lowlands to the highlands in China. We studied 97 Chinese R. solanacearum strains that were isolated from four tobacco-growing zones over a wide range of elevations using phylotype specific multiplex polymerase chain reaction (Pmx-PCR) and phylogenetic relationships (egl and mutS). The results showed that all isolates belonged to phylotype I, which were further clustered into eight egl-sequence type groups (egl-group, sequevar): sequevars 13, 14, 15, 17, 34, 44, 54, and 55. In addition, Sequevar 55, found from the highlands, was a new/unknown one. Southeast China (Z3) had the largest number of egl-groups, containing six sequevars. The basin of the Yangzi River (Z1) and southwestern China (Z2) contained five egl-groups. The basin of the Huai River (Z4), near the north of China, where slight bacterial wilt occurred recently, contained a single group, sequevar 15. The distribution of sequevars was associated with elevation. Sequevar 15 was over-represented in lowland elevations, while sequevar 54 and the new/unknown one were only found in areas of moderate to high elevations. This finding suggested that the phylotype I strains infecting tobacco were diverse in China and regional integrated control strategies should be considered.  相似文献   

6.
Real-Time PCR assay was used to quantify the expression of marker genes of the salicylic acid, jasmonic acid and ethylene signaling pathways in seven Solanum lines after inoculation with a Ralstonia solanacearum phylotype I strain, R008. Four Solanum lycopersicum lines (CRA 66, Hawaii 7996, MST 32/1, Quatre carrées), one S. tuberosum line (Spunta), the wild Lycopersicon cerasiforme and Solanum commersonii were used for this investigation. Results revealed very little activation of the jasmonic acid pathway marker genes, lipoxygenase A (LoxA) and protease inhibitor II (Pin2), with no significant difference (p > 0.05) in fold change expression among the Solanum lines. In contrast the salicylic acid pathway marker genes, glucanase A (GluA) and PR-1a, and the ethylene pathway marker genes, osmotin-like (Osm) and PR-1b, were expressed at higher levels with a statistically significant difference (p < 0.05) in fold change expression among the Solanum lines. The resistant lines L. cerasiforme, CRA 66, Hawaii 7996 and S. commersonii showed stronger activation of the salicylic acid and ethylene marker genes than the moderately resistant cultivar (MST 32/1) and the susceptible lines (Quatre carrées and Spunta). The marker genes reached their highest expression levels earlier (4 h.p.i) in the resistant and moderately resistant lines than in the susceptible lines (48 h.p.i.). These results indicate that salicylic acid and ethylene signaling pathways have a significant role in defense against R. solanacearum. The timing and magnitude of the upregulation of gene expression may determine the plant ability to put up a defense response against the pathogen.  相似文献   

7.
The virulence of 29 isolates of Phytophthora infestans collected in potato fields in Hokkaido, Japan, in 2013 and 2014, was tested for race identification. Thirteen different races were identified, each of which had five to eight virulence factors. All of the isolates caused a virulent reaction against plants with R1 and R7, and most of the isolates caused a virulent reaction against plants with R3, R4, R10, and R11. On the other hand, no isolate was virulent against plants with R9. These results demonstrate that the current Japanese P. infestans population is more complex than the population in the 1990s from the viewpoint of race.  相似文献   

8.
Tomato root exudates were analyzed using a bioassay to detect the chemoattractant for Ralstonia solanacearum. An activated charcoal-adsorbed fraction of root exudates from tomato cultivar Oogata-fukuju had chemoattractant activity for R. solanacearum strain MAFF 730138. The active component, purified using a Sep-Pak C18 cartridge, an activated charcoal column, diol-modified silica gel, and NH2-modified silica gel, is a new hydrophobic attractant. The final purified fraction produced a single peak in a diol-modified silica gel HPLC analysis.  相似文献   

9.
Rhizoctonia cerealis causes sharp eyespot in cereals and the pathogen survives as mycelia or sclerotia in soil. Real-time Polymerase Chain Reaction (qPCR) assays based on TaqMan chemistry are highly suitable for use on DNA extracted from soil. We report here the first qPCR assay for R. cerealis using TaqMan primers and a probe based on a unique Sequence Characterised Amplified Region (SCAR). The assay is highly specific and did not amplify DNA from a range of other binucleate Rhizoctonia species or isolates of anastomosis groups of Rhizoctonia solani. The high sensitivity of the assay was demonstrated in soils using a bulk DNA extraction method where 200 μg sclerotia in 50 g of soil were detected. DNA of the pathogen could also be amplified from asymptomatic wheat plants. Using the assay on soil samples from fields under different crop rotations, R. cerealis was most frequently detected in soils where wheat was grown or soil under pasture. It was detected least frequently in fields where potatoes were grown. This study demonstrates that assays derived from SCAR sequences can produce specific and sensitive qPCR assays.  相似文献   

10.
A blood disease pathogenic strain, Ralstonia syzygii subspecies celebesensis was used to study the possible association of biofilm-forming bacteria with the development and severity of blood disease in banana plants. Therefore, the objective of this study was to determine the effects of mono-culture and co-culture inoculation of isolated biofilm-forming bacteria with the blood disease pathogen in banana pseudostems in glasshouse conditions. Putative biofilm-forming bacteria were isolated from an infected banana plant and were further identified using 16SrRNA sequencing. Four isolates, identified as Enterobacter hormaechei, Enterobacter cloacae, Kosakonia radicincitans and Klebsiella pneumoniae, were inoculated as a mono- and co-culture with R. syzygii subsp. celebesensis into 2 months old banana plants. The observation after the 8 weeks of post inoculation showed that plants which were co-inoculated with the pathogen and K. radicincitans, a biofilm-forming bacterium, were the most susceptible towards the infection. In contrast, plants under two treatments (which were co-inoculated with the pathogen and E. cloacae and the pathogen with E. hormaechei) were less susceptible towards the infection. This study revealed the antagonistic effects of two biofilm-forming strains which reduced the severity of infection caused by the pathogenic agent. Scanning electron micrographs of the cross section of plant rhizomes indicated the dissimilarity of adhesion and host colonization conditions of the pathogen in each infected plant from different treatments.  相似文献   

11.
Trichoderma spp. are used as antagonists against different pathogens. Despite many possibilities of using Trichoderma as an antagonist, there are gaps in the knowledge of the interaction between Trichoderma, cassava and Scytalidium lignicola. This fungus causes cassava black root rot and is an inhabitant of the soil, so it is difficult to control. Antagonists may contribute to the possible induction of resistance of plants because, when exposed to such pathosystems, plants respond by producing antioxidative enzymes. The test for potential inhibition of growth of S. lignicola CMM 1098 in vitro was performed in potato-dextrose-agar with two Trichoderma strains T. harzianum URM3086 and T. aureoviride URM 5158. We evaluated the effect of the two selected Trichoderma to reduce the severity of cassava black root rot and shoots. Subsequently, the production of enzymes (ascorbate peroxidase, catalase, peroxidase and polyphenol oxidase) was evaluated in cassava plants. All two Trichoderma strains show an inhibition of the growth of S. lignicola CMM 1098. The most efficient was T. harzianum URM 3086, with 80.78% of mycelial growth inhibition. T. aureoviride URM 5158 was considered the best chitinase producer. All treatments were effective in reducing severity, especially treatments using Trichoderma. Cassava plants treated with T. aureoviride URM 5158 had the highest enzyme activity, especially peroxidase and ascorbate peroxidase. Trichoderma harzianum URM3086 and Trichoderma aureoviride URM 5158 were effective in reducing the severity of cassava black root rot caused by S. lignicola CMM 1098.  相似文献   

12.
In Hokkaido potato fields, tubers produced from the plants with leaf curl symptoms caused by potato leaf roll virus (PLRV) were noted to be more densely covered with Rhizoctonia sclerotia. This observation led us to hypothesize that potato infected with PLRV would have an increased susceptibility to Rhizoctonia solani. To test this hypothesis, in a pot experiment, we inoculated PLRV-infected mother tubers with Rhizoctonia. As a result, PLRV-infected plants produced significantly fewer and smaller tubers than virus-free plants did, suggesting that PLRV-infected plants are more susceptible than virus-free plants to R. solani. Virus-free seed tubers should thus be used to reduce Rhizoctonia diseases.  相似文献   

13.
To elucidate how soft rot disease affects soil characteristics and root-associated, culturable microorganisms in Amorphophallus konjac stands, the responses of soil around roots of A. konjac with soft rot disease were investigated in stands with and without soft rot. Changes in the root-associated culturable microbial community and diversity were investigated by dilution plating. Soil characteristics were compared between stands using standard techniques. A. konjac with soft rot had higher concentrations of available soil P and K, NH4–N, organic matter and water content and lower pH compared with plants without. The community composition of root-associated culturable microorganisms differed between stands with and without soft rot. The microbial community associated with soft rot in A. konjac was characterized by four types of abundant microorganisms (Fusarium solani, Fusarium oxysporum, Pseudomonas chlororaphis subsp. aureofaciens and Stenotrophomonas pavanii) and three types of less-abundant microorganisms (Rhizobium radiobacter, Bacillus thuringiensis and Streptomyces cellulosae), and a small number of Bacillus and Streptomyces species in the rhizosphere and rhizoplane soils. Particular microbial combinations were diametrically opposed between plants with and without soft rot. The richness and diversity of root-associated culturable microorganisms were higher in the stand without soft rot than in the stand with soft rot. A. konjac soft rot led to obvious differences in the diversity and community composition of root-associated culturable microorganisms and in soil characteristics.  相似文献   

14.
The reniform nematodes of the genus Rotylenchulus are semi-endoparasites of numerous herbaceous and woody plant roots and distributed in regions with Mediterranean, subtropical and tropical climates. In this study, we provide morphological and molecular characterisation of three out of 11 valid species of the genus Rotylenchulus: R. macrodoratus, R. macrosoma, and R. reniformis from Greece (Crete), Italy and Spain. The overall prevalence of reniform nematodes in wild and cultivated olives in Greece, Italy, and Spain was 11.5%, 19.0% and 0.6%, respectively. In Greece, R. macrodoratus and R. macrosoma were detected in cultivated olive with a prevalence of 8.2% and 6.2%, respectively, but none of them were found in wild olive. This is the first report of R. macrosoma in Greece. Only one reniform nematode species was detected in olive from Italy and Spain, viz. R. macrodoratus and R. macrosoma, respectively. The parasitism of R. macrosoma on hazelnut in northern Spain was also confirmed for the first time. This study demonstrates that R. macrodoratus and R. macrosoma have two distinct rRNA gene types in their genomes, specifically the two types of D2-D3 for R. macrosoma and R. macrodoratus, the two types of ITS for R. macrodoratus and the testing of the ITS variability in other R. macrosoma populations in different countries. Rotylenchulus macrosoma from Greece and Spain showed differences in nucleotide sequences in the ITS region and D2-D3 of 28S rRNA gene.  相似文献   

15.
Ramularia ranunculicola sp. nov. (Mycosphaerellaceae) is described from Iran on Ranunculus muricatus and compared with other species reported on Ranunculus spp. Ramularia carletonii on Lactuca tuberosa and R. nagornyi on Centaurea solstitialis are newly reported from Iran, and Picris strigosa is a new host for R. picridis (= R. inaequalis s. lat.).  相似文献   

16.
The influence of the squash hybrid RS841 rootstock (Cucurbita maxima x C. moschata) on population dynamics of Meloidogyne incognita and yield of cucumber cv. Dasher II was assessed during 2013 and 2014 in a plastic greenhouse. In addition, the relationship between ecophysiological parameters (plant water status, gas exchange, and leaf reflectance) and Pi and cucumber yield were also estimated in 2013. Nematode densities were determined at the beginning (Pi) and at the end (Pf) of each crop, and the relationship between these parameters was used to estimate the maximum multiplication rate (a), the maximum population density (M) and the equilibrium density (E) per grafted and ungrafted cucumber and cropping season. Moreover, the relationship between the multiplication rate (Pf/Pi) and Pi was compared between grafted and ungrafted cucumber per cropping season. Finally, the relative yield of grafted or ungrafted cucumber was plotted against Pi to determine the tolerance limit (T) and the minimum relative yield (m) by the Seinhorst damage function model. Values of a, M and E in grafted cucumber were higher than in ungrafted one irrespective of the cropping season. These results were supported by comparing the relationship between Pf/Pi and Pi between grafted and ungrafted cucumber. The relationship between Pi and yield fitted the Seinhorst damage function. The values of T and m did not differ between grafted and ungrafted each year. Predawn water potential, net photosynthetic rate, and leaf chlorophyll index decreased with increasing Pi. In addition, relative yield was related to variation in net photosynthetic rate and the leaf chlorophyll index. Under the conditions of this study, RS841 rootstock was neither resistant nor tolerant to M. incognita.  相似文献   

17.
Bacterial pathogens of onion (Allium cepa) plants and their undetected presence in seed can cause substantial losses to onion producers. In this study, 23 Pseudomonas syringae strains were isolated from five onion plants and 18 onion seeds. The symptoms on leaves and seed stalks were irregular lesions with necrotic centres and water soaked margins. The aim of the study was to characterize these P. syringae strains using Biolog GN III carbon source utilization, multilocus sequence typing (MLST) based on partial sequences of four housekeeping genes (cts, gapA, gyrB and rpoD), and to determine whether or not the strains were pathogenic on onion (cv. Granex 33), chive (Allium schoenoprasum cv. Grasiue), leek (Allium porrum cv. Giant Italian) and spring onion (Allium fistulosum cv. Salotte) plants. Both Biolog analysis and MLST analysis separated onion strains into two clusters, one supporting the existence of a new pathovar of P. syringae, and the other corresponding to P. syringae pv. porri. Pseudomonas syringae strains belonging to the new pathovar we pathogenic only on onion plants of the Allium spp. tested. The results of this study revealed that bacterial blight of onion in South Africa is caused by two pathovars of P. syringae sensu lato, namely, the newly described pathovar, allii, and P. syringae pv. porri. The symptoms caused by these two pathovars in the field were indistinguishable.  相似文献   

18.
Phytophthora species are soil-borne pathogens that damage plants in both agro- and natural ecosystems. To suppress the devastating pathogen, we generated a root-specific expression system using a specific promoter (pPRP3) conferring elevated expression of the target gene in roots that are very susceptible to soil-borne pathogens. To verify root-specific expression, we compared β-glucuronidase (GUS) expression driven by a constitutive or root-specific promoters in shoots and roots. In histochemical and fluorometric assays, GUS activity was detected in whole tobacco plants when GUS expression was driven by p35S, but was detected only in the roots by pPRP3. We then expressed a pepper defensin (J1–1) gene in tobacco to elucidate its effect on plant resistance. The accumulation of J1–1 was also tissue-specific in transgenic tobacco plants. Finally, transgenic plants carrying GUS or J1–1 genes in combination with p35S or pPRP3 were inoculated with Phytophthora parasitica var. nicotianae and Pythium aphanidermatum. Disease symptoms were significantly suppressed in transgenic plants that accumulated J1–1, regardless of the promoter used. Furthermore, the expression of PR genes was induced in J1–1 transgenic plants, exhibiting much higher levels in p35S-driven J1–1 plants than in pPRP3::J1–1 plants. These results demonstrated that J1–1 transgenic plants were primed for enhanced expression of PR genes, which provided synergistic effects with the defensin for disease resistance.  相似文献   

19.
Tomato (Solanum lycopersicum L.) ARGINASE2 (ARG2) and THREONINE DEAMINASE2 (TD2) are involved in plant defense. These enzymes act in the midgut of herbivores fed on tomato plants to degrade the essential amino acids Arg and Thr, respectively. Although it has been demonstrated that overexpression of the SlARG2 gene in tomato enhanced its resistance against M. sexta larvae, knock-down the expression of SlTD2 reduced the resistance of tomato to lepidopteran herbivores; it remains unclear whether overexpression of SlTD2 could enhance the resistance of the host plants to herbivores, or whether combined overexpression of SlARG2 and SlTD2 could lead to synergistically enhanced resistance to insects. Here, we generated transgenic Arabidopsis plants overexpressing SlARG2 (SlARG2 OE) and SlTD2 (SlTD2 OE) individually as well as in combination (SlARG2-SlTD2 OE). Overexpression of these genes did not affect Arabidopsis development, seed yield, or Arg and Thr content. Insect-feeding bioassay was performed by feeding diamondback moth (Plutella xylostella L.) larvae on detached leaves of wild-type, SlARG2 OE, SlTD2 OE, and SlARG2-SlTD2 OE plants. Larvae fed on SlARG2 OE leaves showed approximately 31% to 35% reduction in weight and 6% to 10% reduction in survival rate compared to those fed on wild-type leaves. Although larvae fed on SlTD2 OE leaves showed no reduction in survival rate, they gained less weight. Whereas larvae fed on SlARG2-SlTD2 OE leaves showed neither reduction in weight nor reduction in survival rate. We further investigated the arginase enzymatic activity of the SlARG2 OE and SlARG2-SlTD2 OE transgenic plants. The SlARG2 OE line most resistant to diamondback moth larvae displayed the highest arginase activity. Our data indicate that overexpression of SlARG2 or SlTD2 in Arabidopsis can enhance its resistance against diamondback moth, whereas combined overexpression of SlARG2 and SlTD2 did not generate synergistically increased resistance to diamondback moth.  相似文献   

20.
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