Interactions of soil pH,nutrients and moisture on phytophthora root rot of avocado

Summary This experiment employed a factorial design combining 4 soil pH levels, 3 soil moisture levels, with and without the addition ofPhytophthora cinnamomi to the soil to evaluate the conditions that lead to Phytophthora root rot of avocado.An inverse relation between soil pH and leaf production (and root-weight) was observed in nondiseased plants. In soil infested withP. cinnamomi, plant growth and root weights were much depressed by low soil pH, and especially by low soil pH coupled with high soil moisture contents. These interactions were statistically highly significant. Root weights in pots withP. cinnamomi were closely related to the incidence of disease. A disease index was used to visually assess the conditions of roots. Isolation of the pathogen from diseased plant roots confirmed the accuracy of the disease index.A process of elimination suggsts that favorable soil Ca level and not high pHper se was responsible for disease suppression and that the devastating effects of low soil pH was produced by high Mn (and possibly Al) and associated low levels of Ca and P in soil solutions, which led to breakdown of biological control mechanisms.Journal Series No. 2801, Hawaii Institute of Tropical Agriculture and Human Resources.     

Mechanisms of salt tolerance in transgenic Arabidopsis thaliana constitutively overexpressing the tomato 14-3-3 protein TFT7

A hydroponics experiment was conducted to investigate the effects of iron plaque on root surfaces with respect to selenite uptake and translocation within the seedlings of two cultivars of rice (Oryza sativa L. cv Xiushui48 and Bing9652). Different amounts of iron plaque were formed by adding 0, 10, 30, 50, 70 mg Fe l⁻¹ in the nutrient solution. After 24 h of growth, the amount of iron plaque was positively correlated with the Fe²⁺ addition to the nutrient solution. These concentrations of Fe, inducing plaque, had no significant effect on the shoot and root growth of rice plants in 50 μg Se l⁻¹ nutrient solution. The amount of Se accumulated in iron plaque was positively correlated to the amount of iron plaque. Increasing iron plaque decreased the selenium concentration in shoots and in roots. At the same time, the translocation of Se from roots to shoots was reduced with increasing amounts of iron plaque. At both the shorter and longer exposure times, the ratio of root- to-shoot selenium was higher than in the controls. More Se stayed in the roots at the longer exposure time than at the shorter time. The concentration of selenium in the xylem sap was sharply decreased with increasing amount of iron plaque on the rice roots. The DCB (dithionite-citrate-bicarbonate)-extracted Se was up to 89.9–91.1% of the total Se when roots with iron plaque (Fe 70) were incubated in 50 μg Se l⁻¹ solution for 30 min. This DCB-extracted Se, however, accounted for only 21.9–28.7% of total Se when roots with iron plaque were incubated in the same solution for 3 days. Se adsorbed in iron plaque can be desorbed by low-molecular-weight organic acids, similar to the desorption of Se from ferrihydrite. These results suggest that iron plaque might act as a ‘buffer’ for Se in the rhizosphere.     

Containment and spot eradication of a highly destructive,invasive plant pathogen (<Emphasis Type="Italic">Phytophthora cinnamomi</Emphasis>) in natural ecosystems

The invasive plant pathogen Phytophthora cinnamomi (Stramenopila, Oomycota) has been introduced into 15 of the 25 global biodiversity hotspots, threatening susceptible rare flora and degrading plant communities with severe consequences for fauna. We developed protocols to contain or eradicate P. cinnamomi from spot infestations in threatened ecosystems based on two assumptions: in the absence of living hosts, P. cinnamomi is a weakly competitive saprotroph; and in the ecosystems we treated, the transmission of the pathogen occurs mainly by root-to-root contact. At two P. cinnamomi-infested sites differing in climate and vegetation types, we applied increasingly robust treatments including vegetation (host) destruction, fungicides, fumigation and physical root barriers. P. cinnamomi was not recovered at three assessments of treated plots 6–9 months after treatments. Given the high rates of recovery of P. cinnamomi from untreated infested soil and the sampling frequency, the probability of failing to detect P. cinnamomi in treated soil was <0.0003. The methods described have application in containing large infestations, eradicating small infestations and protecting remnant populations of threatened species.     

Distribution of Phytophthora cinnamomi at different spatial scales: When can a negative result be considered positively?

Abstract In October 1999, patches of dead and dying trees were identified in rainforest vegetation throughout the Tully Falls area in north Queensland, Australia. Previous incidents of patch death in the region had been attributed to Phytophthora cinnamomi. The distribution of P. cinnamomi was assessed by testing for its presence in seven sites displaying signs of dieback and seven sites that appeared healthy. Each site was a circular quadrat, 20‐m radius (total area = 1256.6 m²). Within each quadrat, two perpendicular line transects were constructed. A single soil sample (250 g) was taken at the centre point and at 1‐m intervals along each transect. All soil samples were tested for the presence of P. cinnamomi using a combination of lupin baiting, subsequent culturing and microscopic identification. Of the 1134 samples, 783 recorded positive responses. The mean number of positive responses was not significantly greater in patch death sites than in control sites, suggesting that at this scale of resolution the distribution of P. cinnamomi was uniform. However, at spatial scales of 1‐m intervals across transects the distribution of P. cinnamomi was random.     

Current and projected global distribution of Phytophthora cinnamomi,one of the world's worst plant pathogens

Globally, Phytophthora cinnamomi is listed as one of the 100 worst invasive alien species and active management is required to reduce impact and prevent spread in both horticulture and natural ecosystems. Conversely, there are regions thought to be suitable for the pathogen where no disease is observed. We developed a climex model for the global distribution of P. cinnamomi based on the pathogen's response to temperature and moisture and by incorporating extensive empirical evidence on the presence and absence of the pathogen. The climex model captured areas of climatic suitability where P. cinnamomi occurs that is congruent with all available records. The model was validated by the collection of soil samples from asymptomatic vegetation in areas projected to be suitable by the model for which there were few records. DNA was extracted, and the presence or absence of P. cinnamomi was determined by high‐throughput sequencing (HTS). While not detected using traditional isolation methods, HTS detected P. cinnamomi at higher elevations in eastern Australia and central Tasmania as projected by the climex model. Further support for the climex model was obtained using the large data set from south‐west Australia where the proportion of positive records in an area is related to the Ecoclimatic Index value for the same area. We provide for the first time a comprehensive global map of the current P. cinnamomi distribution, an improved climex model of the distribution, and a projection to 2080 of the distribution with predicted climate change. This information provides the basis for more detailed regional‐scale modelling and supports risk assessment for governments to plan management of this important soil‐borne plant pathogen.     

Benzoic acid induces tolerance to biotic stress caused by Phytophthora cinnamomi in Banksia attenuata

Banksia attenuata plants were treated with soil drenches or foliar sprays of benzoic acid (BZA) to determine induced resistance to Phytophthora cinnamomi. Stems of B. attenuata were inoculated with the pathogen 1 week after treatment with BZA. Resistance was estimated by measuring P. cinnamomi lesions on stems. Treatment with 0.10 mM, 0.25 mM or 0.50 mM BZA caused a reduction in lesion size with 0.50 mM BZA applied as a soil drench being the most effective treatment at suppressing the development of lesions. This is the first report of BZA induced host resistance in any plant species to any pathogen.     

Identifying microorganisms which fill a niche similar to that of the pathogen: a new investigative approach for discovering biological control organisms

Understanding the mechanisms of suppressive soils should lead to the development of new strategies to manage pests and diseases. For suppressive soils that have a biological nature, one of the first steps in understanding them is to identify the organisms contributing to this phenomenon. Here we present a new approach for identifying microorganisms involved in soil suppressiveness. This strategy identifies microorganisms that fill a niche similar to that of the pathogen by utilizing substrate utilization assays in soil. To demonstrate this approach, we examined an avocado grove where a Phytophthora cinnamomi epidemic created soils in which the pathogen could not be detected with baiting techniques, a characteristic common to many soils with suppressiveness against P. cinnamomi. Substrate utilization assays were used to identify rRNA genes (rDNA) from bacteria that rapidly grew in response to amino acids known to attract P. cinnamomi zoospores. Six bacterial rDNA intergenic sequences were prevalent in the epidemic soils but uncommon in the non-epidemic soils. These sequences belonged to bacteria related to Bacillus mycoides, Renibacterium salmoninarum, and Streptococcus pneumoniae. We hypothesize that bacteria such as these, which respond to the same environmental cues that trigger root infection by the pathogen, will occupy a niche similar to that of the pathogen and contribute to suppressiveness through mechanisms such as nutrient competition and antibiosis.     

Bioclimatic analysis of the distribution of damage to native plants in Tasmania by Phytophthora cinnamomi

Climatic profiles were generated by the computer program BIOCLIM for three sets of sites in native vegetation in Tasmania: (i) 308 sites at which Phytophthora cinnamomi was isolated from diseased plants Pc+ ive; (ii) 322 sites in healthy plant communities from which P. cinnamomi could not be recovered Pc?ive; and (iii) 801 sites representing the climatic range across Tasmania. A discriminatory analysis comparing the first and third sets indicated that seven of the 16 climatic indices available for analysis were good discriminators of the distribution of damage by P. cinnamomi. The analysis suggests that damage to native vegetation due to P. cinnamomi is unlikely on sites where annual mean temperature does not exceed 7.5°C or annual mean rainfall is < 600 mm. Two maps were produced to indicate those areas of Tasmania that have climates suited to damaging interaction between P. cinnamomi and native vegetation. The first was based on those sites that had annual mean temperature more than 7.5°C and annual mean rainfall less than 600 mm. The second included those sites that matched the values of the Pc+ ive set for all seven good discriminators. The two approaches produced similar results. Areas in which even the most favourable microsites are unlikely to support pathogenic activity by P. cinnamomi constitute less than 20% of the land area. Twelve substantial areas of native vegetation that occur in climates suited to infection by P. cinnamomi. but for which no record of the fungus exists, have been identified.     

Phytophthora cinnamomi in Shanghai and its possible origin

Camellia leaves were most effective for recoveringPhytophthora cinnamomi from soils followed by azalea leaves and cedar needles. A total of 131 isolates ofP. cinnamomi was obtained from soils and roots collected in Shanghai. Among them 125 were A¹ and 6 were A² type. There was little variation in morphological and physiological characteristics among 82 isolates tested. It is suggested that the fungus may have been a recent settler in Shanghai.     

Combined effects of soil properties and Phytophthora cinnamomi infections on Quercus ilex decline

Aims

The importance of soil properties as determinants of tree vitality and Phytophthora cinnamomi root infections was analysed.

Methods

The study comprised 96 declining stands in western Spain, where declining and non-declining holm oak (Quercus ilex L.) trees were sampled. Soil properties (soil depth, Ah horizon thickness, texture, pH, redox potential, soil bulk density and N-NH₄ ⁺ and N-NO₃ ^? concentrations) and P. cinnamomi infections were assessed.

Results

Tree mortality rates increased with low soil bulk densities, which were also associated with more P. cinnamomi-infected trees. Occurrence of infected trees was higher in fine textured soils and in thick Ah horizons. Fine textured soils favoured trees, but with the presence of P. cinnamomi their health status deteriorated. Soil under declining trees had higher N-NO₃ ^?/N-NH₄ ⁺ ratio values than under non-declining trees. Additional soil properties changes associated to grazing were not related to decline and P. cinnamomi infections.

Conclusions

The implications of P. cinnamomi in holm oak decline and the influence of soil properties as contributors to pathogen activity were demonstrated. Fine soil textures and thick Ah horizons, usually favourable for vigour and vitality of trees growing in the Mediterranean climate, were shown to be disadvantageous soil properties if P. cinnamomi was present. Fine soil textures and thick Ah horizons are frequently related with higher levels of soil moisture, which increase the inoculum of the pathogen and favours root infection. Grazing does not seem to be directly linked to Q. ilex health status or P. cinnamomi root rot.     

The Microscopic Examination of Phytophthora cinnamomi in Plant Tissues Using Fluorescent In Situ Hybridization

The microscopic examination of Phytophthora cinnamomi in plant tissues is often difficult as structures such as hyphae, chlamydospores and oospores are frequently indistinguishable from those of other fungi and oomycetes, with histological stains not enabling species differentiation. This lack of staining specificity makes the localization of P. cinnamomi hyphae and reproductive structures within plant tissues difficult, especially in woody tissues. This study demonstrates that with the use of a species‐specific fluorescently labelled DNA probe, P. cinnamomi can be specifically detected and visualized directly using fluorescent in situ hybridization (FISH) without damage to plant or pathogen cell integrity or the need for subculturing. This approach provides a new application for FISH with potential use in the detailed study of plant–pathogen interactions in plants.     

Effect of Brassica Biofumigant Amendments on Different Stages of the Life Cycle of Phytophthora cinnamomi

The oomycete plant pathogen Phytophthora cinnamomi causes a highly destructive root rot that affects numerous hosts. Integrated management strategies are needed to control P. cinnamomi in seminatural oak rangelands. We tested how biofumigation affects crucial stages of the pathogen's life cycle in vitro, in infested soils under laboratory conditions and in planta. Different genotypes of three potential biofumigant plant species (Brassica carinata, Brassica juncea, Brassica napus) were collected at different phenological stages, analysed for their glucosinolate contents, and subsequently tested. The most effective genotypes against mycelial growth and sporangial production were further tested on the viability of chlamydospores in artificially infested natural soils and in planta on Lupinus luteus, a host highly susceptible to P.cinnamomi. Brassica carinata and B. juncea genotypes inhibited mycelial growth, decreased sporangial production, and effectively inhibited the viability of chlamydospores in soil, but only B. carinata decreased disease symptoms in plants. Effective genotypes of Brassica had high levels of the glucosinolate sinigrin. Biofumigation with Brassica plants rich in sinigrin has potential to be a suitable tool for control of oak root disease caused by P. cinnamomi in Spanish oak rangeland ecosystems.     

Development of a dot immunobinding assay to detect Phytophthora spp. in naturally dark-rooted woody plants

An indirect dot immunobinding assay (DIBA) with a PAb raised against an isolate of P. cinnumomi was evaluated to detect Phytophthora spp. in naturally dark-rooted woody plants. Screening of different modifications of the procedure were done with root samples of Chamaecyparis lawsoniana non-infected and infected with P. cinnamomi. With the optimised DIBA procedure, root infection could be diagnosed by a clearly defined halo around the spot and by a colour change in the spot using Fast Red or Fast Blue substrate. Detection of different Phytophthora species in Chamaecyparis plants from commercial nurseries was successful with the optimised procedure.     

Role of Proanthocyanidins in Resistance of the Legume Swainsona formosa to Phytophthora cinnamomi

Reduced flower pigmentation in the legume Swainsona formosa is associated with increased susceptibility to Phytophthora cinnamomi and other soil‐borne pathogens. This study aimed to identify the mechanism for these differences in susceptibility. Chemical analyses of stem tissues that had been previously inoculated with P. cinnamomi revealed that neither anthocyanin nor total phenolic content increased with infection. Such results suggested that observed differences in susceptibility, as indicated by flower colour, were related to preformed rather than induced stem chemistry. Acetone extracts from healthy, uninfected stem tissues of a red‐flowered line were highly toxic to the fungus, while extracts from a white‐flowered line were non‐toxic and those from a pink‐flowered line were intermediate in toxicity and this was correlated with the total phenolic and proanthocyanidin concentration of the extracts. Precipitation of proanthocyanidins with bovine serum albumen removed the toxicity of the extracts. It was concluded that differences in the proanthocyanidin content of tissues contributed to the differences in disease susceptibility of plants with different flower colours.     

The abundance of ground-dwelling invertebrates in a Victorian forest affected by ‘dieback’(Phytophthora cinnamomi) disease

Abstract Phytophthora cinnamomi (cinnamon fungus) is a pathogenic soil fungus that infects plant communities along the southeastern coast of Australia and the southwestern corner of Western Australia. Infection of native plant communities with P. cinnamomi regularly leads to dramatic changes in both the structural and the floristic characteristics of these communities. This study aimed to assess the effect of P. cinnamomi induced changes in plant community attributes upon the abundance and diversity of invertebrates in an area of open forest in the Brisbane Ranges, Victoria. Pitfall trapping was conducted continuously for 1 week within each season over a 3 year period. Invertebrates were sorted to order level, and abundances were compared between times (season and year) and infection status for taxa with normalized distributions (Coleoptera, Collembola, Dermaptera and ants). Non-parametric comparisons were made for other groups (Aranae, Blattodea, Diptera, Hemiptera, Orthoptera, ant ‘morpho-species’ and unidentified larvae) to assess differences on the basis of infection status. Significant differences on the basis of infection were uncommon and, where identified, elevated abundances were more commonly observed at sites infected by P. cinnamomi. Consistent temporal effects (season and year) were observed in normalized data sets. Abundances both within individual taxa and from pooled counts were generally weakly associated with ground-level habitat features. Overall, the impact of P. cinnamomi on vegetative structure and floristics was not reflected in different abundances of ground-dwelling invertebrates.     

Immunological Discrimination of Phytophthora cinnamomi from other Phytophthorae Pathogenic on Chestnut

A polyclonal antiserum (A379) against water soluble proteins from Phytophthora cinnamomi mycelium was produced in rabbit. In ELISA, the 1 : 10 000 diluted antiserum revealed only Phytophthora isolates, not allowing a clear‐cut discrimination among congenerous species, in spite of a generally higher reactivity on P. cinnamomi proteins. The antiserum gave positive reactions in Western blot analyses against mycelial proteins from nine species of Phytophthora and Pythium sp. (grown on rich media), but not with Rhizoctonia solani, binucleate Rhizoctonia, Verticillium dahliae, Fusarium oxysporum and Cryphonectria parasitica. All Phytophthora species showed common epitopes on proteins of molecular masses 77, 66, 51 and 48 kDa. However, a species‐specific protein of 55 kDa was immunodecorated only in P. cinnamomi samples, thus allowing univocal identification of this species. When tested against total proteins from the same fungi grown on water, the antibody revealed diagnostic bands of 55 and 51 kDa in P. cinnamomi only. The antiserum is therefore suitable for the specific identification of P. cinnamomi emerging in distilled water from infected tissues of chestnut, blueberry and azalea.     

Phosphite stimulated histological responses of Eucalyptus marginata to infection by Phytophthora cinnamomi

Phosphite is used to protect plants from the soil borne pathogen, Phytophthora cinnamomi. Although phosphite stimulates resistance to P. cinnamomi, this is the first histological study of its effect on Eucalyptus marginata, an economically important forest tree in Western Australia. Clonal lines of E. marginata, considered resistant and susceptible to P. cinnamomi, were underbark inoculated with P. cinnamomi. 4 days later, they were treated with 0, 2.5, 5 or 10 g L⁻¹ phosphite. Transverse hand sections were stained for suberin and lignin, and histological responses to infection were examined. Defence responses were stimulated at all phosphite concentrations in both clonal lines, and the genotypic difference in lesion length was eliminated within 8 days of treatment. In the resistant line, suberin production was stimulated while in the susceptible line both lignin and suberin were stimulated. By 2 days after treatment, phosphite stimulated a faster rate of suberin production in the resistant line than the susceptible line, but by 4 days after treatment, there was no difference in the increase between the lines. Damage caused by P. cinnamomi was found to extend furthest in the cortex and outer phloem in transverse sections in both genotypes. In the presence of P. cinnamomi, phosphite stimulated mitosis as part of the defence response, with meristematic activity involved in the compartmentalisation of damaged tissue (formation of periderm) and closure of healthy tissue (callus). Phytotoxicity had a detrimental effect in healthy tissues and this was more apparent in the resistant line, where it did not provide the best protection from lesion extension and plant mortality, suggesting phytotoxicity could disrupt defence responses. Phosphite increases the capacity of susceptible and resistant E. marginata clonal lines to wall-off and contain P. cinnamomi colonisation through lignin and suberin deposition, and increased meristematic activity.     

Effect of mycorrhizal status of avocado seedlings on root rot caused byPhytophthora cinnamomi

Summary The mycorrhizal fungusGlomus fasciculatus stimulated growth of avocado seedlings in the presence or absence ofPhytophthora cinnamomi. It appeared thatG. fasciculatus had no effect upon infection byP. cinnamomi or subsequent disease development.     

Potential for dissemination of Phytophthora cinnamomi by feral pigs via ingestion of infected plant material

Feral pigs have long been implicated as potential vectors in the spread of the devastating plant pathogen Phytophthora cinnamomi due to their rooting and wallowing activities which may predispose them as vectors of infested soil. In this study, we aim to determine whether feral pigs have the potential to act as vectors of plant pathogens such as P. cinnamomi through their feeding activity. The typically omnivorous diet of feral pigs may also lead to the passage of P. cinnamomi infected plant material through their digestive system. This study investigates the potential for feral pigs to pass viable P. cinnamomi in their faeces following the ingestion of millet seeds, pine plugs and Banksia leptophilia roots inoculated with P. cinnamomi. Recovery rates of P. cinnamomi from the millet seeds, pine plugs and B. leptophilia roots following a single ingested bolus were 33.2, 94.9 and 10.4 %, respectively supported by quantitative PCR analysis. These results demonstrate that P. cinnamomi remain viable within infected plant material following passage through the pig digestive tract, although the digestive processes reduce the pathogen’s viability. An inverse relationship was observed between the viability of infected material and passage time, suggesting that partially digested plant material provides protection for P. cinnamomi against the adverse environmental conditions of the pig digestive tract. Phytophthora cinnamomi remained viable for up to 7 days in larger pieces of colonised woody plant material such as the pine plugs. A plant infection trial using passaged P. cinnamomi colonised pine plugs showed that even material that remained in the digestive tract for 7 days was capable of infecting and killing healthy plants, susceptible to P. cinnamomi. This study provides compelling evidence that feral pigs have the ability to transport viable P. cinnamomi in their digestive tract.     

Trunk injection of fosetyl‐aluminium controls the root disease caused by Phytophthora cinnamomi on Quercus ilex woodlands

In Spain, Quercus open woodlands are animal ranching systems of organic production seriously threatened by the exotic pathogen Phytophthora cinnamomi. The root disease it causes kills thousands of oaks annually. Effective disease management needs to integrate different techniques, and the use of a resistance inducer such as fosetyl‐Al can play a key role, because the use of potassium phosphite is prohibited in Spain. In a woodland where the pathogen recently arrived, 60 holm oaks in three different defoliation classes (asymptomatic, slight and moderate defoliation) were selected for trunk injection with pressurised capsules containing 4% of commercial fosetyl‐Al or water (controls). Holm oaks were checked periodically for defoliation and presence of the pathogen in roots and rhizosphere soil. Three years after treatments, defoliation was significantly lower in oaks treated with fosetyl‐Al, which even increased canopy cover, in comparison with control oaks, independent of the initial defoliation class considered. Chlamydospore density in rhizosphere soil, as well as the presence of the pathogen into the roots, was not significantly influenced by fosetyl‐Al treatments, although a trend to a lower presence of P. cinnamomi in roots was observed in treated oaks at every soil inoculum density detected. This study has shown that fosetyl‐Al, a phosphonate registered as a fungicide in the European Union, provides protection to holm oaks against P. cinnamomi, even exhibiting a therapeutic effect on pre‐existing infections. Consequently, this effective measure should be considered as part of the integrated approach to control this highly destructive pathogen in holm oak woodlands.