Arbuscular mycorrhizal fungal-induced alteration to root architecture in strawberry and induced resistance to the root pathogenPhytophthora fragariae

Three strawberry cultivars Elsanta, Cambridge Favourite and Rhapsody were inoculated with eitherGlomus fasciculatum orGlomus etunicatum and their growth compared with non-inoculated plants. The roots of all inoculated plants were 55 to 70% colonised after 98 days. Increases in both root and shoot dry weights were measured. Root architecture was also determined and increases in branching were evident in AMF colonised root systems. The remaining plants were then inoculated with the root pathogenPhytophthora fragariae and allowed to grow for a further 58 days before harvest. In two of the cultivars, Cambridge Favourite and Elsanta, AMF reduced root necrosis by approximately 60 and 30% respectively. Only in the least susceptible cultivar, Rhapsody, was no reduction measured in AMF colonised plants. There were differences in the control conferred by the two arbuscular mycorrhizal fungi and this suggests there may be practical benefits of inoculation. Relationships between the presence of roots of different orders, on inoculation with the pathogen, and subsequent necrosis provided a mechanism for identifying root-architecture driven alteration to susceptibility. Root system necrosis was positively correlated with the proportion of the root system made up of higher order roots (3° to 4°) in non-colonised plants and negatively correlated in AMF colonised plants. These data suggest that root-architecture changes are not important per se but factors expressed concurrently may be.     

Effects of Pratylenchus penetrans on development of strawberry wilt caused by Verticillium dahliae

The nematode Pratylenchus penetrans or the closely related P. fallax occurred in three out of four strawberry plantations infested with Verticillium dahliae surveyed in Kent. When plants of cv. Cambridge Vigour were inoculated with P. penetrans and V. dahliae together in field ‘micro-plots’ the nematode increased the rate of wilt development except when the concentration of V. dahliae microsclerotia in the soil was very low. In a pot experiment, under conditions not conducive to symptom expression, the incidence of infection was increased by nematodes in Cambridge Favourite, Cambridge Vigour and Redgauntlet; the incidence was least in Red-gauntlet, the most resistant cultivar. The nematode appeared to cause local changes in the root cortex which aided hyphal penetration of the adjacent tissues. Growth of strawberry plants in the absence of V. dahliae was not significantly affected even by large populations of the nematode, but growth was reduced by V. dahliae at all rates of inoculation in the field. Infection by Verticillium did not affect the total numbers of nematode per plant at the end of the experiments, although the number per g fresh weight was greater on the small amount of white root on severely diseased plants than on the larger amount on healthy or mildly diseased plants.     

Analysis of Cross-Reactive Antigens of Phytophthora fragariae and Strawberry and their Relation to Resistance and Susceptibility

The presence of cross-reactive antigens between five isolates of P, fragariae (Pf 1, Pf 2, Pf 3, Pf 10 and Pf 11) belonging to five physiological races of the fungus and five strawberry cultivars (Cambridge Favourite, Hapil, Red Gauntlet, Siletz and 52AC18) exhibiting different disease responses to the five isolates was demonstrated by Western blotting. Antiserum anti-H, raised against extracts of healthy Cambridge Favourite roots, detected two antigens which were common to all isolates. Concentration of one of these antigens might be related to the pathogenicity of P. fragariae isolates. Antiserum anti-PfM, raised against mycelial suspensions from the five isolates, detected a doublet of 64 and 61 kDa in the soluble extracts of healthy roots from the five cultivars tested. The corresponding root suspensions revealed numerous other antigens which reacted with anti-PfM. In susceptible interactions, the doublet described above and two additional polypeptides of 38 and 31 kda were detected in large concentrations in both the soluble extracts and root suspensions. These four polypeptides were shown to be present in the healthy roots as well as in the mycelial extracts. In resistant interactions, a 116 kDa polypeptide, present in the root suspensions of the healthy host, was detected in the soluble extracts of the infected roots. It is suggested that these antigens might have a role in resistance and susceptibility.     

Phylogenetic clustering of 4 prevalent virulence genes in <Emphasis Type="Italic">Orientia tsutsugamushi</Emphasis> isolates from human patients

The pathogenicity of microbes is involved in many kinds of virulence genes. The relationships between these virulence genes and strains are not clear in Orientia tsutsugamushi yet. In this study, we confirmed the presence of the virulence genes and classified into O. tsutsugamushi isolates using phylogenetic analysis of the virulence genes. We also compared the fatality rates of every isolate via an infection experiment in BALB/c mice using the O. tsutsugamushi isolates, Deajeon03-01, Wonju03-01, and Muju03-01. Moreover, we compared the phylogenetic analysis, in basis with 56 kDa protein sequence which determined from serotype, and virulence genes of O. tsutsugamushi. Our results showed remarkably different fatality rates between Deajeon03-01 and Muju03-01, which are both Boryong strains of O. tsutsugamushi. Also, clustering analyses including these two isolates gave slightly different results depending on whether they were clustered based on virulence genes or on the 56 kDa protein sequences. Consequently, we conclude that fatality rates in O. tsutsugamushi are correlated with differences in both serotypes and virulence genes. We identified some variations within the virulence genes dnaA, virB8, tolR, and trxA among the isolates.     

Evidence of natural selection for disease resistance in Composite Cross Five (CCV) of barley

Cambridge Composite Cross Five (CCV) of barley was studied utilising hordeins, restriction fragment length polymorphisms (RFLPs) and reaction to powdery mildew with a view to understanding the genetic changes occurring in the population. Changes in the frequency of individual hordein patterns as well as pattern combinations showed directional trends in successive generations in three parallel populations maintained as discrete populations since 1977 in Cambridge. Certain hordein pattern combinations were more common in the resistance classes and there was a strong association between hordein patterns and mildew reaction. RFLP analysis revealed that 80% of a random sample taken from generation F₂₄ of Population I had the same restriction pattern as that of the cultivar Algerian, which was one of the original 30 parental lines of CCV. This cultivar is the source of the Mla1 allele in barley improvement programmes in Europe. We argue, based on supporting evidence from hordein analysis and tests of reaction to selected mildew isolates of known virulence isolates together with UK virulence surveys, that selection for Mla1 in Cambridge has been the predominant evolutionary force in CCV in Cambridge.     

The involvement of fungi in the breakdown of sulphited strawberries

Between 8000 and 9000 tons of strawberries are used annually for jam manufacture in the UK, c. 65 % of which are stored in sulphite liquor (6 % SO₂+lime, c. pH 3.0). Sporadic cases of disintegration of sulphited strawberries have been observed increasingly in recent years. In laboratory experiments breakdown of whole berries in sulphite liquor was achieved by including berries partly rotted with Mucor mucedo, Rhizopus sexualis or R. stolonifer, 1.5-2.5 % infected material causing complete breakdown of all of the berries. The inclusion of up to 25 % berries infected with Botrytis cinerea caused no softening of the berries. The addition of culture filtrates to whole fruit in sulphite liquor confirmed observations with the above fungi but also showed that pectolytic enzymes from Aureobasidium pullulans and Trichosporon pullulans could cause breakdown. The incidence of spoilage fungi on commercially harvested fruit indicated that M. mucedo and R. sexualis were the main cause of breakdown, but that A. pullulans increased tenfold on fruit stored at 15 °C overnight before sulphiting. Results from commercially harvested fruit (cv. Cambridge Favourite) in 1975 and 1976 showed that: (I) fruit should be sulphited as soon as possible after harvest, since the storage of harvested fruit prior to sulphiting may give a poorer quality product; (2) there is little difference in breakdown of fruit from plants treated with the pre-harvest fungicides, Elvaron, Mildothane, or Daconil, and (3) fruit grown in different areas of the UK shows varying amounts of breakdown. This may be due to differences in the infection level of Phycomycetes and contamination by other pectolytic fungi or inherent differences in the susceptibility of the fruit to enzyme attack.     

Genotype Analysis of Collagenase Gene by PCR-SSCP in <Emphasis Type="Italic">Vibrio alginolyticus</Emphasis> and its Association with Virulence to Marine Fish

In this article, the genotypes for Vibrio alginolyticus were analyzed based on collagenase gene by PCR-single strand conformation polymorphism (SSCP) method and their relationship to the virulence for marine fish were studied by a bacteria challenging experiment. Results showed total 13 genotypes designated as A1, A2, A3, A4, B, C, D, E, F, G1, G2, H, and I were identified. Among them, isolates from type A1 were dominant in abundance (48.1%) and the isolates from Nanyou had the highest polymorphisms, in which 11 genotypes were identified from 25 isolates. Moreover, the bacteria challenging experiment showed the virulence of V. alginolyticus was correlated to that of the collagenase-SSCP type (r = 0.438, P < 0.01), which demonstrated the strains from type A1, A2, A3, B, C, F, G1, and I were low-virulent isolates, while those from type A4, D, E, G2, and H were high virulence. These data will provide a new understanding of the relationship between virulence of V. alginolyticus and its molecular marker.     

Sources of crown rot (Phytophthora cactorum) infection in strawberry and the effect of cold storage on susceptibility to the disease

Cold-stored plants of strawberry cultivars Tamella, Cambridge Favourite and Redgauntlet were more susceptible to pathogenic isolates of Phytophthora cactorum than similar plants which had not been cold-stored. Indigenous nonpathogenic isolates of P. cactorum did not cause crown rot in cold-stored plants, although a small number of symptomless latent infections occurred. The majority of P. cactorum isolates causing crown rot symptoms were taken from infected strawberry crowns, although two isolates from gooseberry plants, but of uncertain origin, were also pathogenic. Outbreaks of crown rot in areas with no previous history of the disease therefore probably result from the importation of non-indigenous inoculum with planting material. Assessments of the timing of infection in relation to cold storage revealed that a high incidence of death in the cold store and chronic wilt symptoms on planting from the store resulted from initiating symptomless infections prior to cold storage. However, infection during the period immediately after cold storage resulted in rapid wilt symptoms of Phytophthora crown rot. When plated in sterile distilled water for 24 h, pieces of tissue from infected plants which had died during cold storage produced large numbers of sporangia and zoospores. This indicates that such plant material could provide a potent source of inoculum for infections in the post storage thawing environment. It is proposed that a combination of heightened host susceptibility resulting from cold storage and the presence of scatted latent infections or infected debris among the plants could result in a sudden, large scale appearance of crown rot, as sometimes is seen with cold-stored plantings of strawberries.     

Variation in two components of pathogenicity and its relationship with genetic diversity in Plasmopara halstedii (sunflower downy mildew)

The specificity of the two components of pathogenicity: virulence and aggressiveness and its relationship with genetic variability were analysed in a local Plasmopara halstedii (sunflower downy mildew) population. Pathogenic and molecular analyses were carried out on seven isolates including five progeny isolates of five races arising from two parental races 100 and 710. P. halstedii isolates showed significant differences for all aggressiveness criteria and important genetic variations. Three cases of relationship between virulence and aggressiveness for progeny isolates as compared with parental ones were found as positive, negative or uncorrelated. For solving the specificity of these cases, relationship between the two components of pathogenicity among the isolates of three different races localised in the same genetic clade was positive. The hypothesis explaining these cases is discussed.     

The production of strawberry plants from callus cultures

Shoots were regenerated from callus of the commercially important strawberry varieties Bogota, Brighton, Cambridge Favourite, Hapil, Ostara, Rapella, Red Gauntlet and JILA33 which is a promising selection from a current breeding programme.The callus was initiated from explants of petiole or lamina of leaves of micropropagated shoots in vitro or of lamina or peduncle from greenhouse plants. There was more shoot regeneration with callus from lamina than from petiole although with the variety Hapil, regeneration occurred only with callus from peduncle.With seven of the varieties, shoot regeneration occurred on culture media with BAP and 2,4-D whilst with the remaining variety, Cambridge Favourite, it occurred only with medium which contained 1AA- alanine conjugate in place of 2,4-D.Regenerated shoots rooted readily and the plants produced are being studied for somaclonal variation.     

Susceptibility of Strawberry Varieties to Post-harvest Fungal Spoilage

The susceptibility of strawberry fruits from eleven varieties, Cambridge Favourite, Cambridge Vigour, Cambridge Prizewinner, Red Gauntlet, Chanil, Domanil, Grandee, Litessa, Tamella. Senga Gigana and Precosana, to post-harvest spoilage fungi was investigated over the 1974 and 1975 seasons. Although the fruit from the different varieties from each of two sites at each harvest was contaminated with a similar level of fungi, the pattern of development of the spoilage fungi revealed marked differences between some varieties. Botrytis cinerea and Mucor mucedo were the predominant spoilage fungi on all varieties with Rhizopus sexualis and R. stolonifer being of minor importance. The relative importance of these fungi changed on all varieties as the season progressed, although to a different degree according to variety, site and season. Cambridge Vigour, Senga Gigana, and Chanil were the most susceptible varieties, while Red Gauntlet was the most resistant, especially to the Phycomycetes.     

Ultrastructural changes associated with June Yellows in strawberry and with leaf yellowing symptoms of viral and genetic origin in Fragaria, Rubus and Ribes

Light and electron microscopy was used to study the ultrastructural effects of June Yellows (JY) in leaves of strawberry. Four cultivars of strawberry, affected to different extents by JY, were compared with healthy (JY-free) cv. Cambridge Favourite and with strawberry infected with strawberry crinkle rhabdovirus, Fragaria vesca infected with strawberry mottle virus (SMotV), raspberry and black currant showing virus-induced yellowing and with strawberry and raspberry showing chaemeric yellow sectors in the leaves. Except for isometric virus-like particles detected in SMotV-infected F. vesca, no virus-like particles or structures of other pathogenic agents were found in any of the tissues examined. Leaf cells of JY-affected strawberry showed severe disruption of chloroplasts and plasmalemma, whorls of membranous vesicles and decreased vacuole size. The extent and severity of these abnormalities increased with increased severity of JY symptoms but, even in leaves with mild JY symptoms, chloroplast abnormalities were obvious. In the most severely affected leaves, the cells lacked discrete vacuoles and extensive hypertrophy was seen in other organelles such as nuclei and mitochondria. Few, if any, ultrastructural abnormalities were observed in virus-infected strawberry or F. vesca, or in chaemeric leaves of strawberry and raspberry. By contrast, in raspberry and black currant with yellowed leaves caused by virus infection, the cells showed enlarged chloroplasts, decreased vacuole size and vesicle formation. However, chloroplast enlargement and disruption in this material seemed due to increased size of starch grains which were largely absent from swollen chloroplasts of JY-affected strawberry. The ultrastructural abnormalities observed in JY-affected strawberry are, therefore, not inconsistent with the possibility that a pathogenic agent may be involved in the condition.     

The effect of heat treatment and meristem-tip culture on June Yellows in strawberry

Because there is some evidence that June Yellows (JY) of strawberry may be caused by a pathogenic agent, combinations of heat treatment and meristem-tip culture that are known to eliminate some viruses from tissues were used in attempts to cure affected Cambridge Favourite strawberry plants from JY. None of 397 propagants derived from JY-affected plants subjected to various combinations of these treatments were freed from JY. Indeed, all propagants showed more obvious JY symptoms than the parent plants from which they were derived, suggesting that such treatments may be useful for detecting incipient JY in symptomless strawberry stocks.     

Isolation of Eutypa lata from Juglans regia, and Pathogenicity Studies on Different Wood Plants

The fungus Eutypa lata was isolated from diseased walnut trees (Juglans regia) exhibiting small cankers. The morphological characteristics of the culture and the pathogenicity were compared with those of known isolates of the fungus from other hosts. Inoculation tests with walnut isolates on grape, walnut, almond and apricot yielded characteristic cankers. Furthermore, pathogenicity tests on walnut with isolates from other different hosts resulted in differences in virulence. Similar differences in virulence were observed between 13 single ascospore isolates of the fungus ex apricot inoculated on walnut.     

Initial incidence and development of red core of strawberry

The initial incidence of the disease caused by Phytophthora fragariae in healthy stock of strawberry, cv. Merton Princess, was not affected by different systems of cultivation or by no cultivation for several months before preparing infested soil for planting. In disease development studies, healthy plants which were planted in infested soil in April and sampled at 35-day intervals showed symptoms of red core on 1 September. Disease severity (mean % number of diseased roots) increased with time and, when averaged over intervals of 70 days, the greatest increases occurred between the end of July and the beginning of October and between December and February. Whereas the former was due to infection of plants in which the total numbers of new roots were low, the latter was due to an increase in the numbers of diseased roots. Most primary roots were produced in October and early November and 75% of the final total were formed by 15 December. Healthy plants of the cvs Cambridge Favourite and Redgauntlet, dug from the field at monthly intervals and challenged with zoospores, were less susceptible in May and June than in other months. It is suggested that a fungicide applied in late September or October would protect the maximum number of new roots and achieve the optimum effect for a single application.     

A virulent parent with probiotic progeny: comparative genomics of Escherichia coli strains CFT073, Nissle 1917 and ABU 83972

Escherichia coli is a highly versatile species encompassing a diverse spectrum of strains, i.e. from highly virulent isolates causing serious infectious diseases to commensals and probiotic strains. Although much is known about bacterial pathogenicity in E. coli, the understanding of which genetic determinants differentiates a virulent from an avirulent strain still remains limited. In this study we designed a new comparative genomic hybridization microarray based on 31 sequenced E. coli strains and used it to compare two E. coli strains used as prophylactic agents (i.e. Nissle 1917 and 83972) with the highly virulent uropathogen CFT073. Only relatively minor genetic variations were found between the isolates, suggesting that the three strains may have originated from the same virulent ancestral parent. Interestingly, Nissle 1917 (a gut commensal strain) was more similar to CFT073 with respect to genotype and phenotype than 83972 (an asymptomatic bacteriuria strain). The study indicates that genetic variations (e.g. mutations) and expression differences, rather than genomic content per se, contribute to the divergence in disease-causing ability between these strains. This has implications for the use of virulence factors in epidemiological research, and emphasizes the need for more comparative genomic studies of closely related strains to compare their virulence potential.     

Frequency and geographical distribution of specific virulence factors in Bremia lactucae populations in England from 1973 to 1975

Samples of lettuces infected with Bremia lactucae were collected from seven areas of England during 1973-5. These were tested under controlled environmental conditions on a range of differentially resistant cultivars to determine the frequency of virulence (V) factors in the pathogen population. In most cases the results could be accommodated by the relationship advanced by Crate & Johnson (1976). A virulence phenotype was determined for each isolate and individual virulences within isolates were identified as present at high or low frequencies. The most common V factors were V3, V4, V6 and V8 which reflected the resistance (R) factor complement of lettuce cultivars currently in commercial production. There were few gross differences in the geographical distribution of virulence factors between the seven areas. In the West Central region, however, which encompasses the Evesham and Lancashire production areas, results indicated that the use of cultivars containing resistance (R) factors 2 and 8 respectively would have provided protection from the disease during 1974. There were differences in virulence combinations at the holding or locality level within each large area, which could be exploited by growers providing testing was done in their particular locality. Universally susceptible cultivars and those with only limited resistance factors were often found to be infected with B. lactucae populations which contained large numbers of V-factors. It might have been expected that these pathogen populations would have been restricted to cultivars with large numbers of R-factors. This finding conflicts with the suggestion that pathogen populations with complex virulence patterns are ‘unfit’ as compared with those with more simple virulence components.     

Effect of culture substrates on virulence of Beauveria bassiana (Ascomycota: Cordycipitaceae) conidia against the browntail moth,Euproctis chrysorrhoea (Lepidoptera: Lymantriidae)

We studied the effect of different solid substrates on virulence of two Beauveria bassiana isolates against the browntail moth, Euproctis chrysorrhoea (L.) (Lep.: Lymantriidae). Conidia produced on wheat grains, wheat flour, wheat bran, rice flour, rice bran, rice paddy, corn flour, millet, and Sabouraud's dextrose agar with 1% yeast extract (SDAY) as control were compared. There were significant differences among these substrates for their effects on the virulence of produced conidia. Applying 10⁷ conidia/mL of B. bassiana EUT105, produced on rice bran caused the highest (84.9%) and on rice flour, the lowest (57.6%) mortalities. Bioassay on fifth-instar larvae using aerial conidia harvested from wheat grains, rice paddy, and SDAY indicated that conidia from wheat grains had the highest virulence while those from rice paddy, the lowest.