The Low-Temperature-Induced Viable-But-Nonculturable State Affects the Virulence of Ralstonia solanacearum Biovar 2

ABSTRACT The physiology and virulence of Ralstonia solanacearum biovar 2 strain 1609, kept in water at 4 and 20 degrees C, were studied. At 20 degrees C, total cell and plate count (colony forming units; CFU) numbers were similar, between log 5.03 and log 5.55 CFU, and log 5.03 and log 5.51 cells per ml, at days 0 and 132, respectively. However, CFU in the cultures kept at 4 degrees C dropped from log 6.78 CFU/ml at day 0 to below detection after 84 days. The presence of catalase in the agar resulted in higher CFU, and at day 84, log 1.95 CFU/ml still was detectable. No colonies were observed at day 125. The presence of viable-but-nonculturable (VBNC) cells in the 4 degrees C cultures was confirmed using SYTO9 viability staining. Viable cell numbers were log 1.77 higher than CFU on plates with catalase. At day 84 and after 125 days, log 3.70 viable cells per ml still were present. Shifts in subpopulations differing in viability were found by flow cytometric sorting of 4 degrees C-treated cells stained with SYTO9 (healthy) and propidium iodide (PI; compromised). The SYTO9-stained cell fractions dropped from 99 to 39%, and the PI-stained fractions increased from 0.7 to 33.3% between days 0 and 125. At 20 degrees C, the SYTO9-stained fraction remained stable at 99% until day 132. SYTO9-stained cells sorted from 4 degrees C cultures at day 100 were injected into tomato plants. Upon incubation for 30 days, these plants did not show wilting. However, more than log 4.19 CFU and log 8.17 cells were recovered from these plants. Cells from colonies isolated from the nonwilted plants did not regain their virulence as demonstrated by subsequent injection into several new sets of tomato plants. Cells from 4 degrees C cultures injected at day 125 were not able to cause wilting of, or proliferate in, tomato plants. The threat posed by VBNC R. solanacearum cells upon incubation at 4 degrees C was thus ephemeral because cells lost their capacity to cause disease after 125 days.     

Ralstonia (Pseudomonas) solanacearum Race 3 (Biovar 2) in Surface Water and Natural Weed Hosts: First Report on Stinging Nettle (Urtica dioica)

The population dynamics of the brown rot bacterium Ralstonia (Pseudomonas) solanacearum in surface water of two selected water-areas were monitored over a two-year period. In some cases during summer, high bacterial numbers (up to 10⁶ cfu l^–1) were observed. In a host plant survey a few plants of stinging nettle (Urtica dioica) were found to be a natural host of the bacterium when plants were growing with their roots in contaminated water. The significance of U. dioica in the epidemiology of the brown rot bacterium is not yet known and subject to further investigation. Pathogenicity of R. solanacearum to stinging nettle (U. dioica) and bittersweet (Solanum dulcamara) was demonstrated in a greenhouse experiment.     

Limited survival of Ralstonia solanacearum Race 3 in bulk soils and composts from Egypt

Survival of Ralstonia solanacearum race 3 biovar 2 (phylotype II sequevar 1) in Egyptian soils and compost was studied under laboratory and field conditions. Survival of the pathogen under laboratory conditions varied with temperature, water potential and soil type, with temperature being the major determinant of survival of the pathogen. The effects of temperature and moisture content were variable between different experiments, but survival was generally longer at 15°C than at 4, 28 and 35°C respectively. Survival was also longer when moisture levels were constant compared with varying moisture levels at all temperatures. In experiments to compare the effects of progressive drying in sandy and clay soils there was a difference in survival times between the two soil types. In sandy soils, the pathogen died out more rapidly when soil was allowed to dry out than in controls where the soil was kept at constant water potential. In clay soils there was little difference between the two treatments, possibly due to the formation of a hard impermeable outer layer during the drying process, which retarded water loss from within. Survival in mature composts at 15°C was of the same order of magnitude as in soils but shorter at 28°C, possibly owing to increased biological activity at this temperature, or a resumption of the composting process, with concomitant higher temperatures within the compost itself. The maximum survival time recorded over all soil types and conditions during in vitro studies was around 200 days. In field studies, the maximum survival time in both bare sand and clay was around 85 days at depths up to 50 cm. The survival time was reduced in field experiments carried out in summer to less than 40 days and in one study when the ground was flooded for rice cultivation, the bacterium could not be detected 14 days after flooding. The maximum survival time of R. solanacearum in infected plant material or in infested soil samples incorporated into compost heaps was less than 2 weeks. At the culmination of field soil and compost experiments, no infection was detected in tomato seedlings up to 10 weeks after transplanting into the same soils or composts under glasshouse conditions at a temperature of 25°C.     

Development of Specific Recombinant Monoclonal Antibodies Against the Lipopolysaccharide of Ralstonia solanacearum Race 3

ABSTRACT Recombinant single-chain antibodies (scFvs) against the lipopolysaccharide of Ralstonia solanacearum (biovar 2, race 3) were successfully selected by phage display from a large combinatorial antibody library. Characterization with regard to cross-reaction and use in routine immunoassays showed that the selected antibodies had improved characteristics when compared with the polyclonal antiserum that is currently used for brown rot diagnosis of potato in the Netherlands. The isolated monoclonal scFvs reacted in both enzyme-linked immunosorbent assay (ELISA) and immunofluorescence cell staining with all race 3 strains tested, but with only some strains belonging to other races. Furthermore, only a few cross-reactions with saprophytic bacteria, which also cross-reacted with polyclonal antisera, were observed. Using ELISA, one of the recombinant antibodies detected as few as 5 x 10(3) bacteria in potato tuber extracts. Therefore, this antibody is potentially useful for detection of R. solanacearum race 3.     

Immunofluorescence Colony-staining (IFC) for Detection and Quantification of Ralstonia (Pseudomonas) solanacearum Biovar 2 (Race 3) in Soil and Verification of Positive Results by PCR and Dilution Plating

A procedure was developed for specific and sensitive quantitative detection of Ralstonia (Pseudomonas) solanacearum biovar 2 (race 3) in soil. It is based on immunofluorescence colony-staining (IFC) followed by confirmation of the identity of fluorescent colonies by PCR-amplification or dilution plating on a semi-selective medium, SMSA. Addition of sucrose and the antibiotics cycloheximide and crystal violet to the non-selective trypticase soy broth agar resulted in increased colony size and staining intensity of R. solanacearum in IFC. Verification of IFC-results by picking cells from IFC-positive colonies followed by dilution plating of the suspended cells on SMSA was highly efficient. The success rate was 92% and 96% with spiked and naturally contaminated soils respectively. Several other bacterial species which cross-reacted with polyclonal antibodies in IFC also grew on SMSA and were difficult to distinguish from R. solanacearum, thereby necessitating confirmation of the results. Rapid verification of IFC-positive results directly by PCR-amplification with primers D2/B specific to division 2 of R. solanacearum had a success rate of 86% and 96% with spiked and naturally contaminated soil samples, respectively. Primers D2/B reacted with all R. solanacearum division 2 strains, and strains of R. syzygii and the banana blood disease bacterium, but not with saprophytic bacteria cross-reacting in IFC with R. solanacearum antibodies. In comparative tests, IFC was able to detect consistently ca. 100 cfu g^–1 of soil, a detection level similar to that found with direct plating on SMSA, but less laboriously, whereas detection level with a bioassay on tomato plants was only 10⁴–10⁵ cfu g^–1 of soil.     

Survival of Ralstonia solanacearum Biovar 2, the Causative Agent of Potato Brown Rot, in Field and Microcosm Soils in Temperate Climates

ABSTRACT After outbreaks of potato brown rot in three different fields in the Netherlands, the fate of the brown rot pathogen, Ralstonia solanacearum biovar 2, was monitored in soil by immunofluorescence colony staining (IFC) supported by R. solanacearum division-2 specific polymerase chain reaction. In selected areas of all fields, the R. solanacearum population densities were initially on the order 10(4) to 10(6) per g of topsoil. These population densities then declined progressively over time. In two fields, however, the pathogen persisted for periods of 10 to 12 months. The survival of a selected R. solanacearum biovar 2 isolate, strain 1609, in three soils, a loamy sand and two different silt loam soils, was further studied in soil microcosm experiments. The effects of temperature and soil moisture content were assessed. At 12 or 15 and 20 degrees C, a gradual decline of the population densities was observed in all three soils, from the established 10(5) to 10(6) CFU g(-1) of dry soil to significantly reduced levels, occasionally bordering the limit of detection (10(2) CFU g(-1)of dry soil), in periods of approximately 90 to 210 days. Soil type affected the rate of population decline at 20 degrees C, with the greatest decline occurring in loamy sand soil. In all three soils, the survival of IFC-detectable R. solanacearum 1609 cells at 4 degrees C was severely impaired, reflected in an accelerated decline of CFU counts, to undetectable numbers. Moreover, indications were found for the occurrence of viable but nonculturable strain 1609 cells in the loamy sand as well as in one silt loam soil under these conditions. In addition, a single freezing-thawing cycle caused a significant additional reduction of the culturable R. solanacearum 1609 populations in the three soils, though detectable populations remained. Moderate soil moisture fluctuations of approximately pF 2 did not affect the survival of R. solanacearum 1609 in soil. Severe drought, however, drastically reduced the populations of strain 1609 CFU in all three soils.     

Variation in Genotype and Aggressiveness of Ralstonia solanacearum Race 1 Isolated from Tomato in Taiwan

ABSTRACT A population of Ralstonia solanacearum race 1 from tomato (Lycopersicon esculentum) was analyzed for genetic polymorphism and aggressiveness on tomato. The 46 strains were collected from main tomato-growing areas in Taiwan. Genetic analysis was achieved by two polymerase chain reaction (PCR)-based methods: REP-, ERIC-, and BOX-PCR (collectively as rep-PCR) and random amplified polymorphic DNA (RAPD) techniques. RAPD (with three 10-mers) and rep-PCR revealed 35 and 30 haplotypes, respectively, that were grouped in 14 clusters and 3 clusters, respectively. Distribution of strains into genetic clusters did not appear related to biovar or geographic origin in considering RAPD, rep-PCR, or composite data. Although strains were more dissimilar based on RAPD data than on rep-PCR data, the two techniques gave complementary results for strain clustering. A set of 40 strains representing the main haplotypes was inoculated on six tomato cultivars differing in their bacterial wilt resistance. Six groups differing in general level of aggressiveness and cultivar specificity were detected. Although populations were highly diverse in both genotype and aggressiveness, no association was found between the two characteristics. Although the sample sizes in this study were not adequate to draw definite conclusions about population structure, these results will be valuable for future population genetic studies on R. solanacearum.     

Diversity of Brazilian biovar 2 strains of Ralstonia solanacearum

Ralstonia solanacearum is responsible for bacterial wilt disease. Specific and accurate identification of this pathogen is essential for protection of susceptible crops as well as breeding resistant varieties. Historically, R. solanacearum has been classified into biovars based on the use of sugar and alcohol as carbon sources, into races based on its ability to infect different hosts, more recently into phylotypes based on the intergenic transcribed sequence of the ribosomal RNA genes 16S and 23S and into sequevars based on the endoglucanase gene (egl) sequence. Race 3 biovar 2 (R3Bv2) is widespread in South and Central America, and in Brazil it is present in all potato-producing regions as the most prevalent strain. In this study, we classified 53 Brazilian R. solanacearum biovar 2 (Bv2) strains by traditional and molecular methods. PCR with specific primers confirmed all 53 bacterial strains as belonging to the R. solanacearum species complex, and all were classified as biovar 2A or 2T based on acidification of sugars and alcohols. Multiplex phylotype PCR assigned all strains to phylotype II. Phylogenetic analysis of egl sequences showed that most Bv2 strains from Brazil analyzed in this study did not cluster with known sequevars and are less clonal than the R3Bv2 strains reported for other countries. This is the first study to address the diversity of a collection of Brazilian R. solanacearum strains using the phylotype and sequevar classification scheme.     

Überleben von Ralstonia solanacearum Biovar 2 (Rasse 3), dem Erreger der Schleimkrankheit der Kartoffel, in Boden und auf verschiedenen Materialien (Mikrokosmos)

Microcosm studies were carried out to test the survival of Ralstonia solanacearum biovar 2 (race 3) in soil at the permanent wilting point (wp) water content and at field capacity (fc) water content and on various material. Soils were placed at permanent ?5°C, 4°C, 15°C and 20°C and weekly fluctuating ?10/0/+10°C and the material at 5, 15 °C, 20°C with relative humidity (rh) uncontrolled or at constant 10% or 90%. In soil, survival was clearly dependent on temperature independent of water content. At 20°C Ralstonia solanacearum could be reisolated up to 364 days, at 15°C up to 290 days, at 4°C up to 209 days and at fluctuating temperatures (?10/0/+10°C) only up to 18 days. The lower the temperature, the more the population declined. At 15°C and 20°C appr. 10⁷ cfu/g soil were detected after 100 days, whereas at ?5°C only 10² cfu/g soil were detected after only 18 days. The pathogen was longer detectable in sandy-clay loam than in lighter sandy soil. It could be longer reisolated at wilting point and the populations did not decline as rapidly as at field capacity. Ralstonia solanacearum could best survive on material surfaces like rubber, plastic and varnished metal with maximum survival of 40 days at 5°C and 10% rh. In general there is a low risk of Ralstonia solanacearum overwintering under European climatic conditions when the fields are cleared of plant debris and the soil is frozen. Contamined material surfaces pose the risk of pathogen transmission to healthy tubers.     

Survival of Ralstonia solanacearum and Ralstonia pseudosolanacearum in drain water

The survival in drain water of two strains of Ralstonia solanacearum and three strains of Ralstonia pseudosolanacearum, including two strains able to cause wilt in roses, was determined. Water draining from drip‐irrigated rock wool mats on which roses were grown was supplemented with the pathogen and survival was monitored at 4, 12, 20 and 28°C for up to 112 days. All strains were able to survive for at least 112 days in drain water at 12 and 20°C, but at 4°C maximum survival was 56 days. At 28°C, the survival period was strain dependent, but was at least 56 days. Populations declined gradually in non‐sterile drain water to a low level (maximum 100 cfu mL^?1 after 112 days). In sterile drain water (autoclaved prior to addition of populations), no or only a limited decline in populations was found at 112 days, dependent on strain and temperature. Drain water that tested negative for Ralstonia in the dilution plating assay was tested for the presence of cells in a viable but non‐culturable state (VBNCs). Tomato plants were inoculated, but no symptoms developed, and plants sampled 22 days post‐inoculation were negative in a plating assay. Therefore, no indications were found that VBNCs were present.     

7种杀菌剂对姜瘟病菌Z-AQ-2菌株的抑菌活性

青枯菌4号生理小种引起的姜瘟病是我国生姜生产上的巨大障碍。本研究以分离自5种不同寄主的8株青枯菌为研究对象,采用最小抑制浓度法(MIC)评价了它们的铜抗性水平,发现分离于山东安丘的4号小种Z-AQ-2菌株的铜抗性水平最高,为1.6mmol/L;采用生长速率法测定了7种常用杀菌剂对Z-AQ-2菌株的室内毒力,结果表明:3%中生菌素可湿性粉剂对姜瘟病菌的抑菌活性最高,其EC50为5.2mg/L,其次为6%寡糖·链蛋白可湿性粉剂,EC50为21.2mg/L,20%叶枯唑可湿性粉剂在3种不含铜的农药中EC50值最高,为58.7mg/L,4种铜制剂抑菌效果相对较差,EC50为51.5~148.7mg/L;进一步采用Colby法评价了叶枯唑与氢氧化铜、叶枯唑与中生菌素以及中生菌素与氨基寡糖素的混用效果,结果表明:三组药剂的混用均可产生增效或加成效果。     

Comparative behavior of Ralstonia solanacearum biovar 2 in diverse plant species

Ralstonia solanacearum causes bacterial wilt in numerous plant species worldwide. Although biovar 2 mostly affects solanaceous crops, identification of new hosts remains a matter of concern since there is still no clear-cut distinction between host and nonhost plants. In this work we provide data based on histological studies on the status of 20 plant species, most of them of potential interest in crop rotation. Plants were watered with a beta-glucuronidase-expressing derivative of R. solanacearum biovar 2, and after a month of incubation, sections of roots and stems were analyzed to localize the pathogen on surface, in cortex and/or xylem. Depending on whether the xylem was colonized or not, plants were classified as hosts or nonhosts, respectively. Hosts generally affected in a few xylem vessels or occasionally in all xylem bundles were classified as tolerant. These included some cabbage, kidney bean, and rutabaga cultivars, and the weed bittersweet nightshade (Solanum dulcamara). Nonhosts were the cultivars tested of alfalfa, barley, black radish, carrot, celery, colocynth, fennel, fiber flax, field bean, field pea, horseradish, maize, and zucchini. However, barley and maize, though nonhosts, may act as reservoirs for the pathogen. The present work constitutes a basis for further studies on cropping systems in fields where R. solanacearum has been detected.     

Anaerobic disinfestation of tare soils contaminated with Ralstonia solanacearum biovar 2 and Globodera pallida

Tare soil is soil attached to harvested products like potato tubers. Tare soil becomes a considerable waste stream after storage, washing and processing of harvested products. There is a high risk on contamination of tare soils with (quarantine) phytopathogens, because of import of harvested products from different regions of the world. Disinfestation is necessary to make tare soils applicable for agricultural production. Anaerobic (non-chemical) soil disinfestation (ASD) and inundation are suitable methods to disinfest tare soils. Two different soils (marine loam and glacial sand) were either treated with five (2011) or four (2012) treatments and these were: (i) no treatment (control), (ii) freshly cut grass, (iii) Herbie 7025, (iv) inundation (five cm water on top of the soil surface), and (v) combination of Herbie 7025 and inundation (2011 only). Containers with treated and untreated soils were inoculated with two quarantine phytopathogens, Ralstonia solanacearum biovar 2 and Globodera pallida. After soil inoculation, the containers were airtight closed and only opened for destructive sampling after 84 days. Then, soils were analysed for the presence of R. solanacearum biovar 2, using immunofluorescence colony staining, and for the presence of viable G. pallida eggs, determined by the number of juveniles hatched from eggs and lured to potato root exudate. Strong (> 99.4 %) declines of both pathogens in treated versus the appropriate control soils were observed. Repetition of the experiment revealed the same pattern and therefore it was concluded that ASD and inundation, as separate treatments, have promise for remediation of tare soils from contaminating quarantine pathogens.     

Comparison of Ethylene Production by Pseudomonas syringae and Ralstonia solanacearum

ABSTRACT Strains of Pseudomonas syringae pv. pisi and Ralstonia solanacearum produced ethylene at rates 20- and 200-fold lower, respectively, than strains of P. syringae pvs. cannabina, glycinea, phaseolicola, and sesami. In the current study, we investigated which ethylene biosynthetic pathways were used by P. syringae pv. pisi and R. solanacearum. Neither the activity of an ethylene-forming enzyme nor a corresponding efe gene homolog could be detected in R. solanacearum, suggesting synthesis of ethylene via 2-keto-4-methyl-thiobutyric acid. In contrast, 2-oxoglutarate-dependent ethylene formation was observed with P. syringae pv. pisi, and Southern blot hybridization revealed the presence of an efe homolog in this pathovar. The efe genes from P. syringae pvs. cannabina, glycinea, phaseolicola, pisi, and sesami were sequenced. Nucleotide sequence comparisons indicated that the efe gene in pv. pisi was not as highly conserved as it was in other P. syringae pathovars. The pv. pisi efe homolog showed numerous nucleotide substitutions and a deletion of 13 amino acids at the C-terminus of the predicted gene product. These sequence alterations might account for the lower rate of ethylene production by this pathovar. All ethylene-producing P. syringae pathovars were virulent on bush bean plants. The overlapping host range of these pathovars suggests that horizontal transfer of the efe gene may have occurred among bacteria inhabiting the same host.     

PM 7/21 (2) Ralstonia solanacearum,R. pseudosolanacearum and R. syzygii (Ralstonia solanacearum species complex)

Specific scope

This Standard describes a diagnostic protocol for Ralstonia solanacearum, Ralstonia pseudosolanacearum and Ralstonia syzygii, i.e. phylotype/sequevar strain in the Ralstonia solanacearum Species Complex (RSSC). 1 It should be used in conjunction with PM 7/76 Use of EPPO diagnostic protocols.

Specific approval and amendment

Approved in 2003‐09. First revised in 2018‐02.     

壳聚糖对青枯劳尔氏菌生长及其生物膜形成的影响

为建立快速检测壳聚糖抑菌效果的方法,研究其对青枯菌生物膜形成的影响并探索其抑菌机理,利用酶标仪检测壳聚糖与菌悬液混合培养物的吸光值,计算抑菌率,同时利用结晶紫染色法对壳聚糖处理后青枯菌生物膜形成能力进行评价,并进行透射电镜观察.结果表明,在壳聚糖浓度低于0.15 mg/mL时,随浓度增加其抑菌效果亦显著增加,其浓度超过0.15 mg/mL之后,其抑菌效果达到同一水平,最高抑菌率为74.3％;而壳聚糖浓度对青枯菌生物膜形成能力未产生显著影响;透射电镜观察结果显示:壳聚糖通过破坏青枯菌细胞膜抑制细菌生长.     

Ralstonia solanacearum Biovar 2/Rasse 3 (Erreger der Schleimkrankheit der Kartoffel) in Oberflächengewässern und Solanum dulcamara – Ergebnisse eines mehrjährigen Monitorings in Bayern

The potato brown rot, caused by the bacterium Ralstonia solanacearum biovar 2/race 3 (potato race) was classified as quarantine pathogen by the European Union (EU) due to the risks it would pose for the cultivation of potatoes and tomatoes. Quarantine regulations stipulate control surveys and tests on potatoes used as seed, for food as well as industrial purposes and in surface water to check for contamination with R. solanacearum. Toward this end, the Institute for Plant Protection of the Bavarian State Research Center for Agriculture has been conducting an intensive survey for the presence of the potato brown rot pathogen in Bavarian rivers since 1997. An important component of this monitoring is the testing of potential weed hosts growing near river banks. Every year, from June to September, water- and plant samples have been collected from rivers in potato cultivation areas and near potato processing plants. Since the start of the survey, a total of 70 rivers has been tested. The presence of the pathogen was checked by immunofluorescence antibody staining (IFAS) and polymerase chain reaction (PCR). Positive IFAS or PCR tests were checked again by biotests and pathogenicity tests on eggplant (Solanum melongena). So far R. solanacearum could be found in nine Bavarian rivers. Concerning wild plants growing near the river banks the bacterium could be isolated only from roots of the bittersweet nightshade (Solanum dulcamara). However, from stinging nettle (Urtica dioica), nodding beggartick (Bidens cernua), black nightshade (Solanum nigrum), great yellowcress (Rorippa amphibia) and gipsywort (Lycopus europaeus) R. solanacearum could not be isolated. In contaminated rivers, with S. dulcamara growing on the river banks, R. solanacearum could be detected repetitively for several years. In 1997 it was found that rivers can be contaminated by the pathogen, if insufficiently treated sewage from potato processing plants is released into waterways. Potato cultivation is threatened by surface water only, if contaminated water is used to irrigate potato fields. Consequently, surface water should never be used for irrigation of potato fields or application of plant protection products.     

姜瘟病菌拮抗放线茵的筛选与鉴定

从山东各地区姜田随机采集土样30份,利用稀释分离法分离到放线菌菌株621株.采用琼脂移块法测定这些菌株对姜瘟病菌(Ralstonia solanacearum)的拮抗作用,共筛选出具有拮抗作用的放线菌菌株53株.选择抑菌活性最强的菌株Y7进行进一步研究.菌株Y7孢子丝顶端呈小螺旋形或钩状,孢子椭圆形至圆柱形.在5种供试培养基中,菌株Y7只在燕麦培养基上产生黄色色素.该菌株能利用麦芽糖、葡萄糖、果糖、L(+)树胶醛糖、半乳糖和山梨糖,不能利用蔗糖、木糖、肌醇、纤维二糖和山梨醇.形态特征、生理生化特征和16S rDNA序列分析结果表明,Y7为淡紫灰链霉菌(Streptomycetes lavendulae).正交实验结果表明其最佳发酵条件是小米培养基pH 7.0,28℃,培养14 d.     

广东茄科青枯菌致病力分化及其DNA多态性分析

 分别采自广州、增城、东莞、花都、三水、清远、电白、高要8个菜区的番茄、茄子和辣椒上的31个青枯病菌株,经人工接种于10个鉴别寄主植物上,结果表明,它们的致病力存在明显的差异。聚类分析这31个菌株,可以聚为3个组:第I组菌株主要来自种植番茄和茄子历史较长的广州、东莞、增城老菜区,其致病力较强;第Ⅱ组菌株主要分离自茄子和辣椒上,其致病力中等;第Ⅲ组菌株主要来自近年来新发展的三水市各番茄产区,它们的致病力较弱。从200个随机引物中筛选出17个引物用于上述31个菌株DNA的RAPD分析,共扩增出523条带,其中468条为多态性带,占89.5%。聚类分析这31个菌株,又可聚为4个簇群:第I簇群主要分离自已推广种植多年的丰顺、金丰等抗病番茄品种上;第Ⅱ簇群分离自抗病的番茄新品种新星、年丰和石碣紫红茄上;第IV簇群主要分离自辣椒和茄子;而第Ⅲ簇群来源包括番茄、茄子和辣椒这3种寄主植物。上述试验结果说明,广东茄科青枯菌的致病力存在明显的分化现象,其DNA存在较高的遗传多样性。