Physiological, Biochemical and Molecular Analyses of an Italian Collection of Agrobacterium tumefaciens strains

Physiological, biochemical and molecular characteristics of Agrobacterium tumefaciens strains isolated in Italy from different host plants were analysed. Diseased plants were collected from several nurseries located in nine different regions. Out of 1293 strains isolated from 12 fruit tree and six ornamental plant species, a group of 120 strains was chosen as representative of the whole collection. The majority of the strains were biovar 2 (82.5%), agrocin 84 sensitive, and were isolated from stone fruit trees. Most of the strains identified as biovar 1 were isolated from ornamental plants and were insensitive to A. radiobacter antagonistic strain K84. Some strains that were isolated from Euonymus spp, Prunus GF 677 and Pyrus communis (pear) OHF tumours could not be allocated to any of the three Agrobacterium biovars. PCR-restriction fragment length polymorphism of the rrs gene plus the intergenic spacer was used for strain fingerprinting and characterisation. Results showed a wide genetic variability within the biovar 1 strains and homogeneity within the biovar 2 group. Biovar 2 strains from Sardinia were highly variable and differed from the biovar 2 strains isolated from the other regions of Italy.     

Serological Differences among Erwinia amylovora Biovars

Serological properties were compared among four biovars of Erwinia amylovora. Two biovars (bvs. 1 and 2) from Maloideae sources outside of Japan, one (bv. 3) from Rubus idaeus and one causing bacterial shoot blight of pear (bv. 4) had different reactions in Ouchterlony double diffusion tests using living cells and lipopolysaccharides (LPS) as antigens. These findings indicate that E. amylovora can be classified into three serotypes; i.e., bvs. 1 and 2, bv. 3 and bv. 4. From results of Ouchterlony double diffusion tests and immunoblots, the specific antigen of the three serotypes may each exist in the LPS of E. amylovora strains. Received 27 March 2002/ Accepted in revised form 2 July 2002     

Rhizogenic Agrobacterium biovar 1 and cucumber root mat in the UK

An unusual root disease of cucumbers has affected an increasing number of hydroponic crops in the UK since 1993. Symptoms include upward growth of roots, root proliferation sufficient to distort the shape of rockwool cubes and slabs, thickened roots and occasionally reduced root production and an increase in bent fruit. Rhizogenic Agrobacterium biovar 1 has previously been associated with the problem. In a survey of 37 nurseries in 1997, rhizogenic Agrobacterium bv. 1 isolates were obtained from 17 of 22 nurseries with symptoms and only occasionally from nurseries without symptoms. Young cucumber plants inoculated with rhizogenic Agrobacterium bv. 1 isolates, previously obtained from cucumber roots with symptoms, developed typical symptoms. The survey data and pathogenicity test support rhizogenic Agrobacterium bv. 1 as the primary causal agent of root mat in the UK. Although Agrobacterium bv. 1 isolates that do not harbour an Ri-plasmid are considered avirulent, their abundance in rockwool slabs as potential vectors of the Ri-plasmid may be a major factor in the epidemiology of the disease.     

Diversity of Ralstonia solanacearum causing potato bacterial wilt in Iran and the first record of phylotype II/biovar 2T strains outside South America

The diversity of 40 strains of Ralstonia solanacearum causing bacterial wilt of potato in the major potato-growing areas of Iran was assessed. Based on rep-PCR genomic fingerprinting, strains fell into two distinct groups. The first group contained 37 of the 40 strains and the second consisted of three strains from a narrow tropical region in Iran. The three strains from the narrow tropical region were found to be phenotypically and genotypically most similar to R. solanacearum biovar 2T strains, whereas all other strains were phenotypically and genotypically identified as being R. solanacearum biovar 2/race 3. Phylogenetic analysis of endoglucanase gene sequence information of two of the strains from the tropical region revealed that they belonged to phylotype II of the R. solanacearum species complex and had 100% sequence similarity to a biovar 2T strain from potato in Peru. This is the first report of the presence of R. solanacearum phylotype II/biovar 2T in Iran and the first report of the existence of this group of R. solanacearum outside South America.     

Host specificity and genetic diversity of race 4 strains of Ralstonia solanacearum

Ralstonia solanacearum race 4 isolates were obtained from Zingiberaceae plants in India during bacterial wilt outbreaks. Polyphasic phenotypic and genotypic analysis revealed intraracial diversity and dominance of biovar 3 over biovar 4. Biovar 3 strains were isolated from very severely wilted Zingiberaceae plants in the field and found to be present across diverse geographical, host and seasonal boundaries. It was hypothesized that these isolates belong to a single, ‘fast wilting’, lineage. Using one ‘fast wilting’ isolate in controlled inoculations, rapid wilt was observed in ginger within 5–7 days. Wilting was also observed in several other closely and distantly related hosts such as turmeric (Curcuma longa), aromatic turmeric (Curcuma aromatica), black turmeric (Curcuma caesia), sand ginger (Kaempferia galanga), white turmeric (Curcuma zeodaria), awapuhi (Zingiber zerumbet), greater galangal (Alpinia galanga), globba (Globba sp.), small cardamom (Elettaria cardamomum) and large cardamom (Ammomum subulatum) of the Zingiberaceae family, and in tomato (Solanum lycopersicum). Molecular analysis, including multiplex PCR‐based phylotyping, sequence analysis of 16S rDNA, 16–23S intergenic spacer and the recN gene, and multilocus sequence typing, revealed minimal differences between fast wilting isolates, confirming that almost all belong to the same lineage. Biovar 4 was isolated from plants showing slow wilt progression and self‐limiting wilting in restricted geographical locations instead, and was identified to be genetically distinct from the fast wilting biovar 3 isolates. To the authors' knowledge, this is the first report of host range and genetic analysis of R. solanacearum race 4 in India.     

Impact of kiwifruit bacterial canker on productivity of cv. Hayward kiwifruit using observational data and multivariable analysis

A virulent strain of Pseudomonas syringae pv. actinidiae biovar 3 (Psa), which causes bacterial canker in kiwifruit, was first recorded in New Zealand in November 2010. This strain has severely affected Actinidia chinensis var. chinensis ‘Hort16A’ kiwifruit productivity but its effect on green Actinidia chinensis var. deliciosa ‘Hayward’ kiwifruit productivity has been variable. An observational study design was used to develop explanatory models to quantify the impacts of Psa infection on productivity (tray equivalents per hectare) of Hayward kiwifruit harvested in 2012, using data captured by industry from 2599 orchards. A total of 934 orchards were Psa positive at the end of the study period. Multivariable linear regression was used to model 2012 productivity in the presence of Psa, while controlling for regional differences, elevation, 2011 productivity, harvest dates and application of agrichemicals. The model showed productivity was initially higher in the presence of Psa, and was not reduced until after 1 year of infection. The relationship between protective spray use and productivity was also quantified. It is likely that improved disease management has offset the impact of the disease and future research should consider a reassessment of the effects of disease after longer term exposure to Psa in New Zealand. The use of an observational cohort study to assess disease impacts using multivariable analysis could have wider application in the field of plant epidemiology.     

Suppression of Ralstonia solanacearum in soil following colonization by other strains of R. solanacearum

The effect of prior colonization of a sterile loam soil and a sterile clay loam soil by individual soil bacteria on the subsequent growth of a bacterial wilt pathogen Ralstonia solanacearum YU1Rif43 (tRNA type III: Seal et al. 1992: Appl. Environ. Microbiol., 58, 3759–3761) was investigated. Various strains, belonging to the same type, the same species, the same genus, Gram-negative, Grampositive, or fungi, were used. The degree of suppression of the growth of R. solanacearum YU1Rif43 was markedly different depending on the species that had previously colonized the soil, hereafter referred to as priorcolonists. All the strains belonging to R. solanacearum type III suppressed the growth of R. solanacearum YU1Rif43 markedly, while strains of R. solanacearum type I and type II showed a moderate suppressive effect on R. solanacearum YU1Rif43. The suppressive effect of the strains belonging to species other than R. solanacearum, including fungal strains, was relatively limited, or some strains did not show any suppressive effect. The production of bacteriocin did not appear to be related to the strong suppressive effect of the R. solanacearum type III strains. Possible mechanisms for the suppressive effect of priorcolonists on R. solanacearum YU1Rif43 are discussed in relation to nutrients and physical sites in soil available for growth.     

福建青枯雷尔氏菌的遗传多样性及其生防放线菌的筛选

为了明确福建青枯雷尔氏菌（简称青枯菌）的遗传多样性,综合菌株的演化型、生化型及基于内源葡聚糖酶基因egl的序列变种鉴定,对福建省8个地区的番茄、辣椒和茄子寄主分离的56株青枯菌进行分析。结果表明：供试的56株青枯菌均属于演化型Ⅰ;53株为生化型Ⅲ（占94.64%）,1株为生化型Ⅱ,2株为非标准生化型;从序列变种来看,4株来自茄子的青枯菌均属序列变种15,24株来自辣椒的青枯菌中,23株属于序列变种14,1株为序列变种16,28株番茄青枯菌鉴定出7个序列变种。进一步,选择上述鉴定的生化型Ⅲ和生化型Ⅱ的代表菌株为靶标菌进行生防菌筛选。结果表明,供试14株放线菌中,筛选到1株对生化Ⅲ青枯菌有拮抗作用的放线菌FJAT-31535。基于菌落形态特征和16S rRNA基因序列相似性分析,菌株FJAT-31535属于链霉菌属（Streptomyces sp.）。     

广西烟草青枯菌菌系及其主要生理特性研究

６５个烟草青枯菌菌株均分离自广西的２１个产烟县（市）。全部菌株鉴定属于茄科假单胞。对它们的生化变种、生理小种类型及其他一系列生理特性进行了比较测定。研究结果表明,广西烟草青枯菌的菌系类型较为单一,除３个菌株不能利用甘露醇,暂定为生化变种Ⅲ－１外,其余均属典型的生化变种Ⅲ。根据几种鉴别植物寄主和烟叶组织注射渗透的测定结果,所有菌株也均属生理小种１号。其他１５种生理特性测定结果,除菌株的耐盐能力有一定     

Nitrate alters the flavonoid profile and nodulation in pea (Pisum sativum L.)

A rhizosphere application of NO _inf3 ^sup- and/or naringenin affected the Pisum sativum — Rhizobium leguminosarum biovar viciae symbiosis. NO _inf3 ^sup- (5 mM) lowered while naringenin raised the nodulation status (nodule numbers and weight) and nodule efficiency (C₂H₂ reduction activity). However, the inhibitory effect of NO _inf3 ^sup- was to some extent alleviated when applied in combination with naringenin. The plant biomass was increased by the application of NO _inf3 ^sup- and naringenin, either alone or in combination, while a higher root: shoot ratio was observed only in the naringenin-treated plants. Root flavonoids are known to regulate the expression of nod genes; their high-performance liquid chromatography profile was influenced in different ways by NO _inf3 ^sup- and naringenin.