不同渗透压及pH环境对烟草青枯病菌致病力的影响

为探究不同渗透压和pH环境对烟草青枯病菌Ralstonia solanacearum致病力的影响,用Biolog PM 9～10代谢板中96种渗透压和96种pH环境培养烟草青枯病菌,并采用穿刺法接种于烟草离体叶片,测定不同环境下烟草青枯病菌对烟草的致病情况。结果表明,烟草青枯病菌可致病的渗透压范围包括1%～2%氯化钠、2%～3%硫酸钠、5%～20%乙二醇、1%甲酸钠、2%尿素、1%乳酸钠、20～100 mmol/L磷酸钠、10～100 mmol/L硫酸铵、10～100 mmol/L硝酸钠及10～20 mmol/L亚硝酸钠。可致病pH范围为5.0～8.0;当pH 4.5时,烟草青枯病菌在分别与L-正缬氨酸和5-羟色氨酸共培养时均可致病,与其余33种氨基酸共培养时则均不能致病;当pH 9.5时,烟草青枯病菌在与所有35种供试氨基酸共培养时均不能致病;烟草青枯病菌在葡萄糖苷、辛酸盐、半乳糖苷等10种化合物培养下均可致病。表明渗透压和pH环境会严重影响烟草青枯病菌的生长和致病力。     

Specific Detection of Biovars of Ralstonia solanacearum in Plant Tissues by Nested-PCR-RFLP

A sensitive and specific assay, based on a Nested-PCR-RFLP protocol, was developed for the detection of biovars of Ralstonia solanacearum, the causal agent of bacterial wilt. Oligonucleotide primer pairs were selected within the hrp gene region. Specific amplification of the hrp fragments was obtained for all R. solanacearum strains and also for two closely related species, Pseudomonas syzygii and the blood disease bacterium. No amplification was observed for a wide range of other bacterial species, including R. pickettii and Burkholderia cepacia. Digestion with HindII provided four distinct restriction profiles specific to biovars or groups of biovars of R. solanacearum: one for biovar 1 strains originating from the Southern part of Africa, one for American biovar 1 and biovars 2 and N2 strains, one for biovars 3 and 4 strains, and one for biovar 5 strains. When applied to either pure culture or infected plant tissues, Nested-PCR allowed detection as low as 10³cfu ml^–1, which corresponds to 1cfu per reaction. Amplification was partially or completely inhibited by compounds contained in plant extracts (potato plant and potato tuber, tomato, tobacco, eggplant, pepper and Pelargonium asperum). A combined PVPP/BSA treatment prior to amplification permitted reliable Nested-PCR detection of R. solanacearum strains in plant samples. Nested-PCR-RFLP, assessed with isolates from Reunion Island but also applicable to any R. solanacearum strain, provides a wide range of possible uses for identification, detection and epidemiological investigations.     

烟草青枯病菌与其拮抗菌解淀粉芽胞杆菌的代谢表型差异分析

为探究解淀粉芽胞杆菌X60作为烟草青枯病生防菌剂的潜力,采用Biolog代谢表型技术比较了2种细菌的不同代谢表型。结果表明,烟草青枯病菌和解淀粉芽胞杆菌分别能代谢19%、41%的碳源,43%、77%的氮源,95%、86%的磷源以及100%、69%的硫源,分别有94、91种生物合成途径,49、95种渗透压表型以及19、94种pH代谢表型;解淀粉芽胞杆菌比烟草青枯病菌代谢显著的碳源有L-果胶糖、D-甘露糖等34种,氮源有腺苷、胞苷等29种;烟草青枯病菌比解淀粉芽胞杆菌代谢显著的碳源有D-糖二酸、半乳糖醇等9种,氮源有缩二脲、葡萄糖苷酸等11种;解淀粉芽胞杆菌的渗透压和pH环境适应力比烟草青枯病菌强;解淀粉芽胞杆菌具有脱羧酶和脱胺酶的活性。研究表明,2种细菌的代谢表型间存在较大差异,解淀粉芽胞杆菌的碳源、氮源、渗透压及pH代谢表型较烟草青枯病菌的丰富,烟草青枯病菌的磷源、硫源和生物合成途径代谢表型较解淀粉芽胞杆菌的丰富。     

我国长江流域和南方地区花生青枯菌遗传多样性分析

为明确不同青枯菌的遗传多样性和其在花生植株上的致病力差异,采用国际上新的青枯菌演化型分类模式,对从我国长江流域和南方地区9个花生种植区分离的95株花生青枯菌Ralstonia solanacearum菌株进行遗传多样性分析,基于内源葡聚糖酶基因egl对青枯菌进行系统发育研究,并对供试青枯菌的致病力进行测定。结果表明,所有95株菌株均属于青枯菌演化型I型,即亚洲分支类型。在序列变种分类上,所检测的9个花生种植区中有8个种植区的花生青枯菌菌株属于序列变种14,仅有1个种植区(广西壮族自治区贺州市)的花生青枯菌菌株属于序列变种48,表明我国长江流域和南方地区花生青枯菌群体遗传多样性水平较低。青枯菌致病力测定结果表明,来自赣州市的菌株GZ-1、贺州市的菌株HZ-2和宜昌市的菌株YC接种到花生植株14 d后,花生的病情指数分别为43.8、75.0和87.5,而来自其它6个花生种植区的菌株接种花生后,其病情指数均为100.0,表明菌株GZ-1和HZ-2的致病力较弱,而其它7个花生种植区代表性菌株的致病力均较强。     

Implementation of an artificial reaction control in a TaqMan method for PCR detection of <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> race 3 biovar 2

A previously published TaqMan PCR test for R. solanacearum race 3 biovar 2 was modified to enable both the validation of negative results and the confirmation of positive results in a closed-tube system. Negative results were validated through the use of a reaction control plasmid, designated pRB2C2, which was designed to generate a 94bp product using the same amplimers targeting the primary diagnostic 68bp sequence in R. solanacearum race 3 biovar 2 DNA. SYBR Green was included in the reaction mix to facilitate the identification of post-reaction products using melt peak analysis. The 94bp reaction control had a melt peak temperature of about 90°C, while the diagnostic target amplicon had a melt peak temperature of about 83°C; thus positive results could be easily confirmed and distinguished from the reaction control product. Addition of pRB2C2 at 100 copies per reaction had no effect on the sensitivity of the TaqMan assay for R. solanacearum race 3 biovar 2, and the modified assay successfully detected R. solanacearum race 3 biovar 2 in infected, asymptomatic tomato stems and leaves as well as in potato tubers and stems.     

Molecular identification of some African strains of Ralstonia solanacearum from eucalypt and potato

Ralstonia solanacearum is a known bacterial pathogen of eucalypt and potato plants in Africa. A survey was undertaken to detect this pathogen in eucalypt plantations in South Africa, the Democratic Republic of Congo, and Uganda. Numerous bacterial strains were isolated from trees with symptoms typical of bacterial wilt, but only seven were positively identified as R. solanacearum. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, based on the hrp (hypersensitive response and pathogenicity) gene region was used to determine and group the biovars of these R. solanacearum strains. The eucalypt isolates and one potato isolate formed a biovar 3 cluster, whereas the two other potato isolates formed a cluster that corresponded to biovar 2. Amplified fragment length polymorphism (AFLP) analysis confirmed these clusters. Therefore, PCR-RFLP can be used as a reliable diagnostic technique to enable researchers to rapidly identify the pathogen.     

Phylogenetic relationships of Ralstonia solanacearum species complex strains from Asia and other continents based on 16S rDNA, endoglucanase, and hrpB gene sequences

The 16S rDNA, endoglucanase, and hrpB genes were partially sequenced for Asian strains of Ralstonia solanacearum spp. complex, including 31 strains of R. solanacearum and two strains each of the blood disease bacterium (BDB) and Pseudomonas syzygii. Additional sequences homologous to these DNA regions, deposited at DDBJ/EMBL/GenBank databases were included in the analysis. Various levels of polymorphisms were observed in each of these DNA regions. The highest polymorphism (approximately 25%) was found in the endoglucanase gene sequence. The hrpB sequence had about 22% poly-morphism. The phylogenetic analysis consistently divided the strains into four clusters, as distinctly shown on the phylogenetic trees of 16S rDNA, hrpB gene, and endo-glucanase gene sequences. Cluster 1 contained all strains from Asia, which belong to biovars 3, 4, 5, and N2. Cluster 2 comprised the Asian strains of R. solanacearum (as biovars N2 and 1) isolated from potato and clove, as well as BDB and P. syzygii. Cluster 3 contained race 3 biovar 2 strains from potato, race 2 biovar 1 strains from banana, and race 1 biovar 1 strains isolated from America, Asia, and other parts of the world. Cluster 4 was exclusively composed of African strains. The results of the study showed the distribution and diversity of the Asian strains, which are present in three of the four clusters. The similarity of Asian strains to those in the other regions was also observed.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession numbers AY464950 to AY465050     

Behavior and mutation of <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> in <Emphasis Type="Italic">Solanum toxicarium</Emphasis> grown in aseptic culture

To study the behavior and mutation of Ralstonia solanacearum in Solanum toxicarium, which is resistant to bacterial wilt, S. toxicarium was grown in aseptic culture and inoculated with R. solanacearum. Although 60%–80% of the inoculated plants were wilting after 2 to 3 days, most wilted plants had recovered by 20 days after inoculation. The pathogen was reisolated from over 98% of inoculated plant stems, but the percentage of recovery decreased the closer the isolation sites were toward the upper stem sections. Three colony types, characterized as fluidal white, nonfluidal red, and a mixture of fluidal white and nonfluidal red, were reisolated from the stems. Nonfluidal red colonies were less virulent on tomato plants than fluidal white colonies.     

Role of organic acids in the mechanisms of biological soil disinfestation (BSD)

Biological soil disinfestation (BSD), or reductive soil disinfestation, achieved by amendment with organic materials such as wheat bran followed by flooding and covering the soil surface, has been used to control some soilborne diseases including Fusarium wilt and bacterial wilt of tomato. During a BSD treatment, accumulation of acetic acid and/or butyric acid was detected with high-performance liquid chromatography. Survival of Fusarium oxysporum f. sp. lycopersici or Ralstonia solanacearum was suppressed by these organic acids. Amendment of these organic acids into soil suppressed the survival of R. solanacearum at lower concentrations than the maximum detected in BSD treatment, indicating that production of these organic acids is one of the mechanisms of control. However, F. oxysporum f. sp. lycopersici in soil survived with the maximum concentrations of these organic acids achieved by BSD; thus, involvement of factors other than organic acids may be involved.     

Occurrence of blood disease of banana in Sumatra,Indonesia

Since 1991, the sudden death of cultivated banana plants has been widely observed in the southern region of Sumatra Island, Indonesia. Wilting from loss of petiole and midrib turgidity, yellowing, and necrosis of leaves was followed by death of the whole plant. Reddish brown bacterial ooze exuded from the cut surface of infected pseudostems and fruits. The colony appearance of the isolated bacterium was similar to that of Ralstonia solanacearum. The bacterium was pathogenic to banana plants but not to tomato. Its bacteriological properties agreed with those of blood disease bacterium (BDB) of banana described previously. The 16S rDNA sequence of strain Banana E had conserved bases characteristic of BDB. Based on these results, the causal agent was identified as BDB, which is a close relative of Ralstonia. The isolates have resistance against antibiotics, such as chloramphenicol and tetracycline.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AB095535     

Autoregulator-dependent Control of Extracellular Polysaccharide Production in Phytopathogenic Bacteria

Extracellular polysaccharides (EPSs) likely provide phytopathogenic bacteria a selective advantage both inside and outside plants. Despite the relatively scant knowledge about EPS biosynthesis in phytopathogenic bacteria, it clearly is a well controlled, complex, energy-intensive process. Unexpectedly, three phytopathogenic bacteria have been found to autoregulate EPS production in response to extracellular signal compounds (pheromones) that they produce. Like many bacteria, Pantoea stewartii subsp. stewartii produces a N-acyl-homoserine lactone (AHL) autoinducer. However, unlike most AHL-dependent autoinduction systems, that in P. stewartii subsp. stewartii somehow represses EPS production in the absence of autoinducer. Instead of an AHL-dependent system (which it also has), Ralstonia solanacearum uses a novel autoregulator identified as 3-hydroxypalmitic acid methyl ester to regulate EPS biosynthesis. A lack of this autoregulator in R. solanacearum results in repression of EPS biosynthesis by a complex two-component sensor/response regulator signal cascade. Xanthomonas campestris pv. campestris has two partially overlapping autoregulatory systems. The autoregulators are incompletely characterized, but one diffusible signal factor (DSF) is thought to be a fatty acid derivative and the other diffusible factor (DF) may be a butyrolactone. The autoregulation pathways in X. campestris pv. campestris are essentially unknown, but EPS production is controlled by both the DSF and DF systems, whereas production of extracellular enzymes and pigment production are regulated independently. In a confined micro-environment, population density and intercellular concentrations of an autoregulator will increase in parallel, so autoregulation is one way that bacteria can coordinate gene expression to synthesize EPS only at high cell density. However, because there is often limited evidence that it is actually cell density that is being detected, researchers should not assume a priori that autoregulation must function for quorum sensing. Some possible reasons for why phytopathogenic bacteria would benefit from delaying EPS production are discussed.     

Small cardamom (Elettaria cardamomum Maton.) and ginger (Zingiber officinale Roxb) bacterial wilt is caused by same strain of Ralstonia solanacearum: a result revealed by multilocus sequence typing (MLST)

Bacterial wilt in cardamom (Elettaria cardamomum Maton) was observed in Kerala state of India. Infected plants showed wilting wherein all leaves roll or curl upward towards the midrib centre, turn yellow, and the whole plant finally dies; the collar region shows water-soaked lesions initially and turns dark brown eventually; copious quantity of bacterial exudate is observed on the cut end of the pseudostem. The bacterium was identified as Ralstonia solanacearum based on a panel of phenotypic characters such as fluidal white colony on Kelman’s medium, biovar assay and biolog assay (BiologGN), and genotypic characters such as Multiplex-PCR based phylotyping, sequences of 16S rDNA, 16-23S intergenic region, and recN gene. Collectively these tests revealed that the R. solanacearum infecting cardamom belong to biovar 3 and phylotype 1 confirming its Asian origin. Upon soil inoculation, the bacterium caused typical wilting of the cardamom plants in three weeks and ginger plantlets in two weeks. Cross transmissibility of the bacterium was observed in cardamom and ginger wherein the plants succumbed to wilt when R. solanacearum from either of the host was inoculated. BOX-PCR fingerprinting revealed that the strain is identical (100%) to a ginger strain of R. solanacearum, which is widely prevalent in the Indian sub-continent. Furthermore, Multilocus Sequence Typing (MLST) based strain comparison confirmed that cardamom and ginger strain were identical to each other at 11 loci. Apart from striking phenotypic and genotypic (allelic) similarities, geographical origin, and cross transmissibility of the cardamom strain of R. solanacearum strongly suggest that the new occurrence of wilt of cardamom in India could have an origin in bacterial wilt of ginger. Perusal of records on Ralstonia-induced bacterial wilt in crop plants, particularly among the Zingiberaceae family, reveals that this is a new report of bacterial wilt disease in small cardamom.     

Phenotypic characterization of Pseudomonas fluorescens PfG32R and its spontaneous gacS mutants and biocontrol activity against bacterial wilt disease of tomato

Pseudomonas fluorescens strain PfG32R actively suppresses the occurrence of bacterial wilt disease of tomato caused by Ralstonia solanacearum. To determine the characteristics affected by spontaneous mutations in the gacS gene, three spontaneous mutants NR1, NR9, and ASW6 were tested for production of several enzymes, antimicrobial activity, and biocontrol activity against bacterial wilt disease of tomato in a greenhouse. Production of biosurfactant and of enzymes such as Tween-80 lipase, gelatin protease, and lecithinase; swarming activity; and antifungal activities against Fusarium oxysporum f. sp. radicis-lycopersici and F. graminearum were inactivated by mutations in the gacS gene, indicating that they are under the regulation of the GacS/GacA two-component regulatory system. Levan production, swimming, antifungal activity against Magnaporthe grisea, and antibacterial activity against R. solanacearum and Erwinia carotovora ssp. carotovora were not controlled by the GacS/GacA system. Biocontrol activities of the three spontaneous mutants varied from strain to strain, indicating that the biocontrol activities of PfG32R are influenced not only by the GacS/GacA system but also by other complex factors, which may not be regulated by the system.     

Identification of a bacterium isolated from galls on carrot and weeds

In 2004, bacterial galls were found on the roots of carrots in Shizuoka Prefecture, Japan. Galls were about 0.1–2 cm in diameter, light brown in color and had rough surfaces. In 2005, similar galls were found on the roots of three weeds: henbit (Lamium amplexicaule L.), Persian speedwell (Veronica persica Poir.) and leaf mustard (Brassica juncea L.). A bacterium that forms white, rough colonies was isolated from the carrot and weeds galls. The bacterial isolates had properties identical with Rhizobacter dauci Goto and Kuwata. Phylogenetic analysis based on 16S rDNA sequences showed that the carrot isolate had the highest homology (similarity of 100%) with that of the type strain of R. dauci. Rep-PCR genomic fingerprinting using BOX A1R primer showed that the carrot and weeds isolates were nearly identical. Pathogenicity of the isolates was confirmed by inoculating the roots of carrots and the weeds. After 2–5 weeks, they formed galls on the roots of the original host species and on other plant species tested. The galls were indistinguishable from those formed naturally, and the inoculated bacterium was reisolated. Thus, the causal bacterium of carrot and weeds gall was identified as R. dauci, and the bacterium was found to have a wider host range than previously known. These weed hosts may serve as inoculum sources for carrot bacterial gall disease.     

Ralstonia solanacearum virulence in tomato seedlings inoculated by leaf clipping

Ralstonia solanacearum is a phytopathogenic bacterium that colonizes the xylem vessels of host plants leading to a lethal wilt disease. Although several studies have investigated the virulence of R. solanacearum on adult host plants, infection studies of this pathogen on the seedling stages of hosts are less common. In a preliminary observation, inoculation of R. solanacearum F1C1 on 6‐ to 7‐day‐old tomato seedlings by a simple leaf‐clip strategy resulted in a lethal pathogenic condition in seedlings that eventually killed these seedlings within a week post‐inoculation. This prompted testing of the effect of this inoculation technique in seedlings from different cultivars of tomato and similar results were obtained. Colonization and spread of the bacteria throughout the infected seedlings was demonstrated using gus‐tagged R. solanacearum F1C1. The same method of inoculating tomato seedlings was used with R. solanacearum GMI1000 and independent mutants of R. solanacearum GMI1000, deficient in the virulence genes hrpB, hrpG, phcA and gspD. Wildtype R. solanacearum GMI1000 was found to be virulent on tomato seedlings, whereas the mutants were found to be non‐virulent. This leaf‐clip technique, for inoculation of tomato seedlings, has the potential to be a valuable approach, saving time, space, labour and costs.     

PCR-based specific detection of <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> race 4 strains

Two primer sets were designed based on the sequence of polymorphic bands that were derived from repetitive sequence-based polymerase chain reaction (rep-PCR) fingerprinting and specifically detected in Ralstonia solanacearum race 4 strains (ginger, mioga, and curcuma isolates). One primer set (AKIF-AKIR) amplified a single band (165bp) from genomic DNA obtained from all mioga and curcuma and some ginger isolates; another set (21F-21R) amplified one band (125bp) from the other ginger isolates. These primer sets did not amplify the bands from genomic DNA of other R. solanacearum strains or of other related bacteria. PCR detection limit for the pathogen was 2 × 10²cfu.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession numbers AB118756 and AB118757     

Analysis of the gacS-gacA regulatory genes of spontaneous mutants of Pseudomonas fluorescens biocontrol strain PfG32R

Pseudomonas fluorescens strain PfG32R, a potential biocontrol agent against soilborne pathogens, frequently loses its antifungal activity and ability to produce enzymes. To characterize genetically the instability of these bacterial functions, we analyzed gacS and gacA genes of PfG32R and three spontaneous mutants of PfG32R (NR1, NR9, and ASW6), which had lost their ability to produce proteases and antifungal activities. The gacS and gacA sequences of PfG32R had 77%–89% and 78%–87% homology, respectively, with several known gacS and gacA homologues. All three spontaneous mutants were subjected to complementation analysis. Introduction of clones containing an intact gacS of PfG32R and another P. fluorescens strain Pf-5 as well as strain tea632 of P. syringae pv. theae complemented all three mutants restoring protease and antifungal activities, indicating a mutation in gacS. In sequencing analysis, the mutants had a deletion or change in amino acids in the conserved sensor kinase domains of GacS. The three mutants maintained both their antibacterial activity against Ralstonia solanacearum and Clavibacter michiganensis ssp. michiganensis and siderophore production, indicating that they are not controlled by the GacS/GacA system. The sequences reported in this article have been deposited in the DDBJ database under accession numbers AB219364 and AB219365.     

<Emphasis Type="Italic">In vitro</Emphasis> Selection of Maize Rhizobacteria to Study Potential Biological Control of <Emphasis Type="Italic">Aspergillus</Emphasis> Section <Emphasis Type="Italic">Flavi</Emphasis> and Aflatoxin Production

The aims of this study were to select bacterial isolates from the non-rhizophere of maize soil and to examine their antagonistic activity against Aspergillus section Flavi strains. The first selection was made through ecophysiological responses of bacterial isolates to water activity (a_w) and temperature stress. Subsequently, an Index of Dominance test (I_D), ecological similarity and inhibition of the lag phase prior to growth, growth rate and aflatoxin B₁ accumulation were used as criteria. From the first assay nine bacterial strains were selected. They grew well at 25 and 30 °C, with growth optima between 0.982 and 0.955 a_W using 48 h of incubation. There was ecological similarity between the bacterial strains Bacillus subtilis (RCB 3, RCB 6), Pseudomonas solanacearum RCB 5, Amphibacillus xylanus RCB 27 and aflatoxigenic Aspergillus section Flavi strains at 0.982 at 25 °C. The predominant interaction between all selected bacteria and fungi in dual culture was mutual intermingling at 0.982. Mutual inhibition on contact and mutual inhibition at a distance was observed at 0.955 a_w, between only four bacteria and some Aspergillus strains. Bacillus subtilis RCB 55 showed antifungal activity against Aspergillus section Flavi strains. Amphibacillus xylanus RCB 27, B.␣subtilis RCB 90 and Sporolactobacillus inulinus RCB 196 increased the lag phase prior to growth and decreased the growth rate of Aspergillus section Flavi strains. Bacillus subtilis strains (RCB 6, RCB 55, RCB 90) and P. solanacearum RCB 110 inhibited aflatoxin accumulation. Bacillus subtilis RCB 90 completely inhibited aflatoxin B₁ accumulation at 0.982 a_W. These results show that the bacterial strains selected have potential for controlling Aspergillus section Flavi over a wide range of relevant environmental conditions in the stored maize ecosystem.     

Bacterial induction of the accumulation of phaseollin,pisatin and rishitin and their antibacterial activity

Bean hypocotyls, pea pods and tomato fruits were tested for phaseollin, pisatin and rishitin production when challenged with the phytopathogenic bacteriaErwinia carotovora, Pseudomonas phaseolicola, P. pisi andP. solanacearum, and their isolated extracellular polysaccharides. All bacteria induced phytoalexin accumulation, whereas only phaseollin and pisatin, but not rishitin, were elicited by EPS. The inhibitory effect of these three phytoalexins on bacterial growth was studied in liquid medium; whereas phaseollin and pisatin strongly inhibited growth, only a slight inhibitory effect resulted from the presence of rishitin in the medium.Samenvatting Bonehypocotylen, erwtepeulen en tomatevruchten werden onderzocht op hun vermogen tot vorming van respectievelijk faseolline, pisatine en rishitine, na inoculatie met de fytopathogene bacteriënErwinia carotovora, Pseudomonas phaseolicola, P. pisi enP. solanacearum en na behandeling met oplossingen van hun extracellulaire polysacchariden (EPS). Alle bacteriesoorten induceerden fytoalexinevorming, terwijl hun EPS wel faseolline- en pisatine-, maar geen rishitinevorming induceerden. Faseolline en pisatine remden de groei van de bacteriën in vloeibaar medium sterk; rishitine daarentegen had slechts een geringe groeiremming tengevolge.     

一株戴尔福特菌NF83-1的鉴定及评价

为研究拮抗菌株NF83-1的分类与生物学特性以及评价其生防能力,通过生理生化特征和16S rRNA基因序列分析对菌株NF83-1进行种属鉴定,以选择性培养基检测其分泌的多种胞外酶,以结晶紫染色法检测其生物膜的形成,采用室内平板对峙生长法测定其抑菌活性,室外盆栽法测定其对番茄立枯病的防效。结果表明,菌株NF83-1被鉴定为戴尔福特菌Delftia tsuruhatensis;菌株NF83-1能产生纤维素酶和嗜铁素,且能在管壁形成较厚的生物膜。菌株NF83-1对立枯丝核菌Rhizoctonia solani、甘蓝黑斑病菌Alternaria brassicae、油菜菌核病菌Sclerotinia sclerotiorum、梨胶孢炭疽病菌Colletotrichum gloeosporioides等病原真菌,梨火疫病菌Erwinia amylovora、番茄青枯病菌Ralstonia solanacearum、水稻细菌性条斑病菌Xanthomonas oryzae pv.oryzicola等革兰氏阴性病原细菌及革兰氏阳性细菌巨大芽胞杆菌Bacillus megaterium都具有明显的抑制作用;菌株NF83-1发酵液原液和100倍稀释液对番茄立枯病的防效分别为48.78%~57.69%和26.83%~36.54%。研究表明,菌株NF83-1是一株具有较好生防应用潜力的拮抗菌株。