Characterisation of pectobacterium wasabiae causing blackleg and soft rot diseases in South Africa

Pectolytic bacteria were isolated from potato tubers and stems showing tuber soft rot and blackleg symptoms. Approximately half (52 %) of the isolates could grow at both 27 and 37 °C while another half (48 %) failed to grow at 37 °C. All isolates could be amplified with primers specific to the pectate lyase (pel) gene. Carbon utilisation profiles could not conclusively identify these isolates. PCR amplification using primers specific for Pectobacterium carotovorum subsp. brasiliensis was positive for all isolates that grew at 37 °C. However, the group that did not grow at 37 °C failed to amplify with P. atrosepticum specific primers. To characterise this group of isolates, the intergenic transcribed spacer region (ITS) was amplified and PCR products digested with two restriction enzymes (RsaI and CfoI) to generate ITS-PCR-RFLP profiles. The profiles of these new isolates were compared to those of the type strains of other pectolytic bacteria. Profiles of five of the selected atypical strains generated with the enzyme CfoI appeared to be most similar to those of P. wasabiae type strain. Phylogenetic analysis using concatenated partial gene sequences of housekeeping genes mdh and gapA clustered these isolates together with those of P. wasabiae reference strains thus confirming their identity. These strains were virulent on potato tubers and stems but did not elicit hypersensitive response on tobacco plants. This is the first report of P. wasabiae causing soft rot and blackleg of potatoes in South Africa.     

Characterization of Pectobacterium carotovorum subsp. carotovorum and brasiliense from diseased potatoes in Kenya

Using a DNA-based typing method, 48 bacterial strains isolated from infected potato (Solanum tuberosum) tubers originating from Kenya were characterized. The pel gene specific primers showed that all the 48 bacterial strains were pectolytic. Subspecies-specific primers EXPCCF/EXPCCR and Br1f/L1r identified 66 % of the strains as Pectobacterium carotovorum subsp. carotovorum while 34 % were identified as Pectobacterium carotovorum subsp. brasiliense based on their characteristic band sizes of 550 and 322 bp, respectively. Amplification of the 16S-23S rDNA (ITS) region did not yield observable differences in banding patterns between the Kenyan strains. However, PCR-RFLP analysis together with partial nucleotide sequences of the housekeeping mdh and gapA genes confirmed the results obtained by the specific primers. Phylogenetic analysis of the concatenated partial gene sequences grouped Pectobacterium carotovorum subsp. carotovorum and Pectobacterium carotovorum subsp. brasiliense Kenyan strains together with those identified in other parts of the world with 90 % and 99 % bootstrap support values, respectively. Pathogenicity assays using representative Kenyan strains demonstrated varied levels of tuber maceration ability. The Pectobacterium carotovorum subsp. carotovorum and Pectobacterium carotovorum subsp. brasiliense Kenyan strains were shown to be less aggressive in causing soft rot when compared to type strains. This study describes for the first time the genetic diversity of pectolytic bacteria causing soft rot disease of potatoes in Kenya.     

Characterization of <Emphasis Type="Italic">Dickeya</Emphasis> and <Emphasis Type="Italic">Pectobacterium</Emphasis> strains obtained from diseased potato plants in different climatic conditions of Norway and Poland

Soft rot and blackleg of potato caused by pectinolytic bacteria lead to severe economic losses in potato production worldwide. To investigate the species composition of bacteria causing soft rot and black leg of potato in Norway and Poland, bacteria were isolated from potato tubers and stems. Forty-one Norwegian strains and 42 Polish strains that formed cavities on pectate medium were selected for potato tuber maceration assays and sequencing of three housekeeping genes (dnaX, icdA and mdh) for species identification and phylogenetic analysis. The distribution of the species causing soft rot and blackleg in Norway and Poland differed: we have demonstrated that mainly P. atrosepticum and P. c. subsp. carotovorum are the causal agents of soft rot and blackleg of potatoes in Norway, while P. wasabiae was identified as one of the most important soft rot pathogens in Poland. In contrast to the other European countries, D. solani seem not to be a major pathogen of potato in Norway and Poland. The Norwegian and Polish P. c. subsp. carotovorum and P. wasabiae strains did not cluster with type strains of the respective species in the phylogenetic analysis, which underlines the taxonomic complexity of the genus Pectobacterium. No correlation between the country of origin and clustering of the strains was observed. All strains tested in this study were able to macerate potato tissue. The ability to macerate potato tissue was significantly greater for the P. c. subsp. carotovorum and Dickeya spp., compared to P. atrosepticum and P. wasabiae.     

Pectinolytic bacteria associated with potato soft rot and blackleg in South Africa and Zimbabwe

Soft rot and blackleg can cause severe economic losses in potato production in South Africa and Zimbabwe depending on climatic conditions. The aim of the study was to identify the predominant bacteria causing potato soft rot and blackleg in these countries. Samples, comprising of stems and tubers from potato plants with blackleg and soft rot symptoms were collected from 2006?C2009 from potato production areas where disease outbreaks occurred. The isolates from these plants and tubers yielded Gram negative, pectinolytic bacteria on crystal violet pectate and inoculated tubers. Identification was based on biochemical and phenotypic characteristics, rep-PCR, Amplified Fragment Length Polymorphisms and sequences of gyrB and recA genes. Isolates from Zimbabwe were identified as Pectobacterium carotovorum subsp. brasiliensis (Pcb) (21 isolates), Dickeya dadantii subsp. dadantii (Dd) (20 isolates), P. c. subsp. carotovorum (Pcc) (16 isolates) and P. atrosepticum (Pa) (4 isolates). Pcb, Pcc and Dd subsp. dadantii were isolated from samples collected from all the regions, while Pa was isolated from Nyanga the coolest region in Zimbabwe. In South Africa, however, Pcb was the most common causal agent of soft rot and blackleg. P. atrosepticum was the only pathogen isolated from samples collected in Nyanga, Zimbabwe, and was not isolated from any South African samples. AFLP analysis separated the Pcb strains into 12 clusters, reflecting subdivision in terms of geographic origin, and Pcc isolates were clearly differentiated from Pcb isolates. A large degree of DNA polymorphism was evident among these 12 clusters. The study identified all the pathogens associated with the blackleg/soft rot disease complex.     

Characterization of Malaysian Pectobacterium spp. from vegetables using biochemical, molecular and phylogenetic methods

In August 2011, vegetable crops showing symptoms of maceration and water soaked lesions on their tuber, leaf, and fruit were collected from four major vegetable growing states in Malaysia including Pahang, Johor, Melaka and Selangor. The majority of the causal organisms isolated from infected tissues (52 strains) were identified as Pectobacterium spp. based on PCR amplification of the pectate lyase (pel) gene and amplification of the 16S-23S rRNA (ITS) with G1 and L1 primers. Physiological and biochemical assays divided Malaysian Pectobacterium species into two main groups: Pectobacterium wasabiae and Pectobacterium carotovorum subsp carotovorum. Partial sequence of PCR product from reaction of putative Pectobacterium spp. with 16S rRNA confirmed the results obtained from physiological and biochemical assays used for identification of the bacterium. Application of specific primers such as Eca1F/Eca2r, Br1f/L1r, EXPCCF/EXPCCR, and also ITS-PCR following by RFLP by restriction enzyme (RsaI) successfully differentiated Malaysian P. wasabiae and P. carotovorum subsp carotovorum isolates from other species and subspecies of Pectobacterium. Phylogenetic analysis of Malaysian isolates with housekeeping genes (mdh, gapA) grouped Malaysian P. carotovorum subsp carotovorum and P. wasabiae in the same cluster with P. carotovorum subsp carotovorum (Ecc380) and P. wasabiae (SCRI488) respectively.     

Characterization of Pectobacterium carotovorum subsp. odoriferum causing soft rot of stored vegetables

Pectobacterium carotovorum subsp. odoriferum has been generally considered to have a narrow host range and has been isolated most often from chicory. Research was conducted to identify 91 Pectobacterium spp. strains isolated from different vegetables in Europe, North and South America, Asia, and Africa, and to compare their ability to cause disease in chicory and potato. Among the 91 strains, 22 strains from Europe were identified as P. c. subsp. odoriferum. Based on phylogenetic analysis of 16S rDNA, recA, and rpoS gene sequences, strains isolated from stored vegetables clustered together with the type strain of P. c. subsp. odoriferum and clustered separately from the P. c. subsp. carotovorum isolates. Eleven strains previously identified as P. c. subsp. carotovorum were reclassified as P. c. subsp. odoriferum. All P. c. subsp. odoriferum isolates were able to cause soft rot symptoms on chicory and potato. Moreover, the symptoms on potatoes were more severe at temperatures from 15 to 37 °C with P. c. subsp. odoriferum isolates than with P. atrosepticum or P. c. subsp. carotovorum isolates. Tissue maceration by P. c. subsp. odoriferum isolates was highest at 28 °C, and at that temperature tissue maceration was two-times greater for P. c. subsp. odoriferum isolates than for P. c. subsp. carotovorum isolates. Symptoms on inoculated chicory leaves were more severe with P. c. subsp. odoriferum (regardless of origin) than with other subspecies or species. To our knowledge, this is the first report that P. c. subsp. odoriferum occurs on a wide range of vegetables and has the ability to cause soft rot during potato storage.     

Characterization of Pectobacterium species from Iran using biochemical and molecular methods

Characteristics of forty strains from macerated potato tubers and water-soaked lesions of some ornamental plants were studied in north parts of Iran. The causal organisms isolated from infected tissues were identified as Pectobacterium spp. based on their physiological and biochemical assays and confirmed by species and subspecies specific PCR and RFLP analysis of 16S–23S intergenic transcribed spacer region. Artificial inoculation of isolates to their related hosts generated the same symptoms on potato and ornamental plants, from which the same bacteria were isolated and identified. We detected two groups of atypical isolates in this study. The first group from potato classified as Pectobacterium carotovorum subsp. carotovorum by phenotypic tests but was unable to elicit HR on tobacco leaves, to grow at 37°C and to amplify the pel gene relevant to this subspecies. The second one from ornamental plants which was again characterized as Pectobacterium carotovorum subsp. carotovorum in biochemical assays, produced a unique ITS-RFLP profile different from all of known Pectobacterium species and subspecies. Our findings based on phylogenetic analysis using concatenated partial sequences of housekeeping genes mdh and gapA, indicated the occurrence of P. wasabiae as a novel species in potato storage in Iran. Furthermore we detected a distinct clade of Pectobacterium spp. from some ornamental plants including Schlumbergera bridgesii, Syngonium podophyllum and Iris spp.     

Virulence of Pectobacterium carotovorum subsp. brasiliense on potato compared with that of other Pectobacterium and Dickeya species under climatic conditions prevailing in the Netherlands

In western Europe, Pectobacterium carotovorum subsp. brasiliense is emerging as a causal agent of blackleg disease. In field experiments in the Netherlands, the virulence of this pathogen was compared with strains of other Dickeya and Pectobacterium species. In 2013 and 2014, seed potato tubers were vacuum infiltrated with high densities of bacteria (10⁶ CFU mL^?1) and planted in clay soil. Inoculation with P. carotovorum subsp. brasiliense and P. atrosepticum resulted in high disease incidences (75–95%), inoculation with D. solani and P. wasabiae led to incidences between 5% and 25%, but no significant disease development was observed in treatments with P. carotovorum subsp. carotovorum, D. dianthicola or the water control. Co‐inoculations of seed potatoes with P. carotovorum subsp. brasiliense and D. solani gave a similar disease incidence to inoculation with only P. carotovorum subsp. brasiliense. However, co‐inoculation of P. carotovorum subsp. brasiliense with P. wasabiae resulted in a decrease in disease incidence compared to inoculation with only P. carotovorum subsp. brasiliense. In 2015, seed potatoes were inoculated with increasing densities of P. carotovorum subsp. brasiliense, D. solani or P. atrosepticum (10³–10⁶ CFU mL^?1). After vacuum infiltration, even a low inoculum density resulted in high disease incidence. However, immersion without vacuum caused disease only at high bacterial densities. Specific TaqMan assays were evaluated and developed for detection of P. carotovorum subsp. brasiliense, P. wasabiae and P. atrosepticum and confirmed the presence of these pathogens in progeny tubers of plants derived from vacuum‐infiltrated seed tubers.     

Genotypic and phenotypic variability of <Emphasis Type="Italic">Pectobacterium</Emphasis> strains causing blackleg and soft rot on potato in Turkey

Pectinolytic bacteria were isolated from 48 potato plants showing the symptoms of blackleg and collected in different fields of commercial potato production areas at Samsun, Amasya, Corum and Yozgat provinces in Turkey in 2015. The survey resulted in the isolation of 26 pectinolytic strains that belonged to P. atrosepticum, P. carotovorum subsp. brasiliense, P. carotovorum subsp. carotovorum and P. parmentieri species based on molecular identification with species-specific PCR and phenotypic characterization. The identified strains indicated typical biochemical characteristics of the assigned species. For 16 representative Pectobacterium isolates 10 unique rep-PCR band patterns were obtained. The 16S rRNA and recA and gapA gene fragment sequencing confirmed the species identity of the isolates. The phenotypic characterization of the strains revealed that for all assays but one (cellulase, protease activity, swimming but not swarming), the tested Pectobacterium species were significantly different from each other proving the diversity of the strains belonging to these genera. Recent outbreaks of blackleg and/or soft rot in potato production areas in Turkey may pose a threat on other crops, as tomato, pepper, cucumber, onion, cabbage, broccoli and sugar beet are cultivated in the same provinces.     

The use of two complementary DNA assays,AFLP and MLSA,for epidemic and phylogenetic studies of pectolytic enterobacterial strains with focus on the heterogeneous species Pectobacterium carotovorum

Amplified fragment length polymorphism (AFLP) markers and multilocus sequence analysis (MLSA) were used to analyse 63 bacterial strains, including 30 soft‐rot‐causing bacterial strains collected from Syrian potato fields and 33 reference strains. For the MLSA, additional sequences of 41 strains of Pectobacterium and Dickeya, available from the NCBI GenBank, were included to produce a single alignment of the 104 taxa for the seven concatenated genes (acnA, gapA, proA, icd, mtlD, mdh and pgi). The results indicate the need for a revision of the previously classified strains, as some potato‐derived Pectobacterium carotovorum strains were re‐identified as P. wasabiae. The strains that were classified as P. carotovorum during the analyses demonstrated high heterogeneity and grouped into five P. carotovorum highly supported clusters (PcI to PcV). The strains represented a wide range of host plants including potatoes, cabbage, avocados, arum lilies, sugar cane and more. Host specificity was detected in PcV, in which four of the six strains were isolated from monocotyledonous plants. The PcV strains formed a clearly distinct group in all the constructed phylogenetic trees. The number of strains phylogenetically classified as subspecies ‘P. c. subsp. brasiliensis’ in PcIV dramatically increased in size as a result of the characterization of new isolates or re‐identification of previous P. carotovorum and P. atrosepticum strains. The P. carotovorum strains from Syria were grouped into PcI, PcII and PcIV. This grouping indicates a lack of correlation between the geographical origin and classification of these pathogens.     

<Emphasis Type="Italic">Pectobacterium carotovorum</Emphasis> subsp. <Emphasis Type="Italic">carotovorum</Emphasis> can cause potato blackleg in temperate climates

It is well established that the pectinolytic bacteria Pectobacterium atrosepticum (Pca) and Dickeya spp. are causal organisms of blackleg in potato. In temperate climates, the role of Pectobacterium carotovorum subsp. carotovorum (Pcc) in potato blackleg, however, is unclear. In different western and central European countries plants are frequently found with blackleg from which only Pcc can be isolated, but not Pca or Dickeya spp. Nevertheless, tubers vacuum-infiltrated with Pcc strains have so far never yielded blackleg-diseased plants in field experiments in temperate climates. In this study, it is shown that potato tubers, vacuum-infiltrated with a subgroup of Pcc strains isolated in Europe, and planted in two different soil types, can result in up to 50% blackleg diseased plants.     

Enhancement of resistance to soft rot (Pectobacterium carotovorum subsp. carotovorum) in transgenic Brassica rapa

Brassica rapa (chinese cabbage) is one of the main vegetable crops grown in Asian countries. Pectobacterium carotovorum subsp. carotovorum causes severe economic loss in this crop as well as in other Brassica crops through soft rot disease. Cysteine proteases like bromelain, papain or ficin show toxic effects to herbivorous insects and pathogenic bacteria. They have been known to be critical factors in plant defence mechanisms. The current study investigated the effect of bromelain gene (BL1) of pineapple (Ananas comosus L. Merrill) on enhancing resistance to soft rot in transgenic Brassica rapa ‘Seoulbaechu’. Three homozygous T₂ lines were inoculated with Pectobacterium carotovorum subsp. carotovorum and BL8-2 line showed the lowest rate of infected leaves (RIL) in both wound inoculation and non-wound inoculation, when the non-infected line showed 100 % RIL in both cases. The highest expression of BL1 gene was also observed in BL8-2 homozygous line. Thus, the over-expressed BL1 gene conferred enhanced resistance to soft rot in Brassica rapa.     

Genetic characterisation of <Emphasis Type="Italic">Pectobacterium wasabiae</Emphasis> causing soft rot disease of potato in New Zealand

Pectobacterium wasabiae has a narrow host range, having previously only been associated with Japanese horseradish. However, recent characterisation of Pectobacterium causing soft rotting in New Zealand has identified putative P. wasabiae isolates pathogenic to potato. In this study, phylogenetic reconstruction of acnA and mdh DNA sequences and fluorescent amplified fragment length polymorphisms (fAFLP) were used to confirm the identity of the putative P. wasabiae isolates. Both methods clustered the potato isolates closely with the type strain for P. wasabiae, ICMP9121, and also differentiated them from other plant pathogenic enterobacteria. PCR, DNA hybridisation and hypersensitive response (HR) assays were subsequently used to investigate the presence in P. wasabiae of the type III secretion system (T3SS) as well as other virulence factors known to be involved in development of disease by enterobacteria. Although all P. wasabiae strains appeared to elicit a type III-dependent HR in tobacco, genes associated with the T3SS and the putative virulence factors HecB and DspE could not be detected. Thus, genetic characterisation of P. wasabiae confirmed that it is a naturally occurring pathogen on potato, which does not possess the same suite of virulence factors that are involved in the pathogenicity of other enterobacteria on this host.     

Soft rot of root chicory in Hokkaido and its causal bacteria

In August 2010, bacterial soft rot was found on root chicory (Cichorium intybus var. sativum) in Hokkaido, Japan. Severely infected plants in fields were discolored, had wilted foliage, and black necrosis of petioles near the crown. Wilted leaves subsequently collapsed and died, forming a dry, brown or black rosette. The root and crown became partially or wholly soft-rotted. Slimy masses on infected areas of roots, turned dark brown or black. Gram-negative, rod-shaped, peritrichously flagellated, facultatively anaerobic bacteria were exclusively isolated from rotted roots, and typical symptoms were reproduced after inoculation with the strains. The bacteria were identified as Dickeya dianthicola, Pectobacterium carotovorum subsp. carotovorum, and Pectobacterium carotovorum subsp. odoriferum based on further bacteriological characterization and the sequence analysis of the malate dehydrogenase gene and 16S rRNA gene. These bacteria should be included with the previously reported Dickeya (=Erwinia) chrysanthemi in Saitama Prefecture, Japan, as causal pathogens of bacterial wilt of chicory.     

Evaluation of a diagnostic microarray for the detection of major bacterial pathogens of potato from tuber samples

Potato can be infected with many bacterial pathogens, the detection of which is necessary in seed certification. In this study, a diagnostic microarray previously tested for specificity of probes for detecting the potato bacteria causing blackleg and soft rot (Pectobacterium atrosepticum, Pectobacterium carotovorum, and Dickeya spp.), ring rot (Clavibacter. michiganensis subsp. sepedonicus), scab (Streptomyces scabies and Streptomyces turgidiscabies) and brown rot (Ralstonia solanacearum) from pure culture was evaluated for analytical sensitivity when testing directly from tuber samples. The microarray readily detected all the bacterial species when 100 ng of the target bacterial DNA from pure culture was mixed with DNA from soil microbes and potato. However, detection was inconsistent when total DNA isolated directly from infected tubers or enriched bacterial culture was used. While the high specificity of the probes could be confirmed from the results of the DNA cocktail experiment used as a control, the study demonstrated that the level of analytical sensitivity of the microarray under the tested condition was not sufficient to detect bacteria directly from tubers. Therefore, in addition to the cost and organizational complexities, the low analytical sensitivity and limited reproducibility of the microarray are constraints for establishing the platform for routine detection of potato bacterial pathogens from tuber samples.     

Pectobacterium spp. associated with bacterial stem rot syndrome of potato in Canada

Pectobacterium atrosepticum, P. carotovorum subsp. brasiliensis, P. carotovorum subsp. carotovorum, and P. wasabiae were detected in potato stems with blackleg symptoms using species- and subspecies-specific polymerase chain reaction (PCR). The tests included a new assay for P. wasabiae based on the phytase gene sequence. Identification of isolates from diseased stems by biochemical or physiological characterization, PCR, and multi-locus sequence typing (MLST) largely confirmed the PCR detection of Pectobacterium spp. in stem samples. P. atrosepticum was most commonly present but was the sole Pectobacterium sp. detected in only 52% of the diseased stems. P. wasabiae was most frequently present in combination with P. atrosepticum and was the sole Pectobacterium sp. detected in 13% of diseased stems. Pathogenicity of P. wasabiae on potato and its capacity to cause blackleg disease were demonstrated by stem inoculation and its isolation as the sole Pectobacterium sp. from field-grown diseased plants produced from inoculated seed tubers. Incidence of P. carotovorum subsp. brasiliensis was low in diseased stems, and the ability of Canadian strains to cause blackleg in plants grown from inoculated tubers was not confirmed. Canadian isolates of P. carotovorum subsp. brasiliensis differed from Brazilian isolates in diagnostic biochemical tests but conformed to the subspecies in PCR specificity and typing by MLST.     

Characterization of <Emphasis Type="Italic">Pectobacterium carotovorum</Emphasis> subsp. <Emphasis Type="Italic">carotovorum</Emphasis> causing soft-rot disease on <Emphasis Type="Italic">Pinellia ternata</Emphasis> in China

Pinellia ternata is a traditional Chinese herb which has been used in China for over 1,000 years. A soft-rot disease characterized by water-soaked lesions and soft-rot symptoms with a stinking odour was commonly observed in cultivated fields of this plant, and Pectobacterium-like bacteria were consistently isolated from the infected tissues. Two typical strains (SXR1 and ZJR1), isolated from Shanxi and Zhejiang, respectively, were identified. Pathogenicity tests revealed that these strains were virulent to P. ternata and induced the same symptoms as observed in the field. Characterization involving fatty acid profile, metabolic and physiological properties, 16S rDNA sequence and PCR-RFLP identified both isolates as P. carotovorum subsp. carotovorum (Pcc). The 16S rDNA of both isolates shared 97–99% sequence similarity with that of Pcc strains. The phylogenetic trees showed that both isolates were clustered in the group of Pcc and P. carotovorum subsp. odorifera and both PCR-RFLP profiles were consistent with the pattern E produced by the minority of Pcc strains. Thus, isolates SXR1 and ZJR1 were characterized as Pcc in spite of some differences. This is the first report that Pcc has been proven as a causal agent of soft-rot disease on P. ternata.     

A pith necrosis caused by Xanthomonas perforans on tomato plants

Pith necrosis is a common disease of tomato in Europe, mainly caused by Pseudomonas corrugata and other soil-borne species of Pseudomonas. During 2011–2012 a survey was conducted in soil-grown tomato crops in southeastern Sicily (Italy). Plants showed pith necrosis, brown discolouration of the vascular tissues, leaf chlorosis and sometimes wilting of leaves. Thirty bacterial isolates from symptomatic tissues, forming colonies on NA and KB, were identified by morphological, biochemical and physiological tests. Among them, seven isolates were analyzed for their 16S rDNA and 16S–23S spacer region sequence that resulted in 99 % identity to that of the Xanthomonas perforans type strain (GenBank accession number GQ46173over 2.085 bp.). Additional sequences of fusA, gapA, gltA, gyrB, lacF, and lepA from one selected isolate were 100% identical to sequences of the Xanthomonas perforans type strain. X. perforans local isolates showed similar genomic patterns with REP-PCR and fAFLP, and were clearly distinguished from other Xanthomonas spp. type strains. In stem-inoculation assays, bacteria isolated from symptomatic tomato plants identified as P. fluorescens, P. putida, P. marginalis, P. citronellolis, P. straminea, and Pantoea agglomerans induced discolouration of vascular tissues, while Pectobacterium carotovorum subsp. atrosepticum isolates induced soft rot. Conversely, the isolates here identified as Xanthomonas perforans were able to induce pith necrosis, vascular discolouration, longitudinal splits and external lesions on stems. This report of X. perforans causing pith necrosis on tomato represents a potentially serious problem that may limit the productivity of tomato crops.     

In vitro antibacterial activity of nanoemulsion formulation on biofilm,AHL production,hydrolytic enzyme activity,and pathogenicity of Pectobacterium carotovorum sub sp. carotovorum

In the present investigation the in vitro activity of nanoemulsion (AUSN1) was evaluated against the plant pathogen Pectobacterium carotovorum sub sp. carotovorum causing the soft rot disease in numerous horticultural crops. With AUSN1 nanoemulsion treatment, minimum inhibitory concentration (MIC) of 0.09 mg ml⁻¹, minimum bacteriostatic concentration (MBC) of 1.30 mg ml⁻¹ and zone of inhibition of 2.90 cm was observed against P. carotovorum. This formulation was able to reduce the bacterial population by 32.2–51.6%, under biofilm formation condition and a complete elimination of population under normal cultivation conditions. When subjected AUSN1 treatment, reduction in biofilm formation, swarming and swimming motility was observed in P. carotovorum strain and also inhibited the N-acyl homoserine lactones (AHL) activity by 13.27–62.3% and AHL production by 32.4–76.13%. Reduction in exo polysaccharide (EPS) synthesis and hydrolytic enzymes production in P. carotovorum strain was observed due to AUSN1 treatment. Reduced hydrophobicity (36.9–56.4%) and adhesion to polystyrene (22.9–47.5%) and potato tuber surfaces (19.5–46.8%) was observed with AUSN1 treatment. All these collective evidences clearly show that, under laboratory conditions, AUSN1 treatment was able to inhibit the soft root incidence in potato tubers.     

萝卜软腐病生防菌株Hy11的筛选、鉴定及定殖特性

为探讨银杏内生细菌对萝卜软腐病的防病作用,通过块根接种和温室盆栽试验筛选对萝卜软腐菌Pectobacterium carotovorum subsp.carotovorum具有高效抑制作用的生防菌株,共获得8株拮抗作用较强的内生细菌,其中菌株Hy11抑菌作用明显.对生防菌株Hy11进行了形态观察、生理生化特性测定、16S rDNA和gyrB基因序列分析,并应用绿色荧光蛋白基因标记菌株Hy11,研究该菌株在萝卜体内的定殖动态.结果显示,该菌株对萝卜软腐病具有较好的防效,对萝卜块根和幼苗的防治效果分别为77.9％和66.7％;对萝卜幼苗的促生率达113.28％.经鉴定,菌株Hy11为解淀粉芽胞杆菌Bacillus amyloliquefaciens.标记菌株Hy11-gfp在喷雾接种银杏叶片后0～3 d种群数量呈急剧下降趋势,10d后保持相对稳定;其在萝卜的根、茎和叶中均能够定殖,在根中的定殖数量最高可达1.2×104 CFU/g.