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.     

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.     

Defining Plant Resistance to Phytophthora cinnamomi: A Standardized Approach to Assessment

Phytophthora cinnamomi is a soil‐borne plant pathogen that causes devastating disease in agricultural and natural systems worldwide. While a small number of species survive infection by the pathogen without producing disease symptoms, the nature of resistance, especially under controlled conditions, remains poorly understood. At present, there are no standardized criteria by which resistance or susceptibility to P. cinnamomi can be assessed, and we have used five parameters consisting of plant fresh weight, root growth, lesion length, relative chlorophyll content of leaves and pathogen colonization of roots to analyse responses to the pathogen. The parameters were tested using two plant species, Zea mays and Lupinus angustifolius, through a time course study of the interactions and resistance and susceptibility defined 7 days after inoculation. A scoring system was devised to enable differentiation of these responses. In the resistant interaction with Z. mays, there was no significant difference in fresh weight, root length and relative chlorophyll content in inoculated compared with control plants. Both lesion size and pathogen colonization of root tissues were limited to the site of inoculation. Following inoculation L. angustifolius showed a significant reduction in plant fresh weight and relative leaf chlorophyll content, cessation of root growth and increased lesion lengths and pathogen colonization. We propose that this technique provides a standardized method for plant–P. cinnamomi interactions that could be widely used to differentiate resistant from susceptible species.     

Survival of Phytophthora cinnamomi and Fusarium verticillioides in commercial potting substrates for ornamental plants

Live plants, particularly when accompanied by soil or potting substrates, are considered the main pathway for international spread of plant pathogens. Modern, rapid shipping technologies for international plant trade increase the probability of plant pathogen survival during transport and the subsequent chances of disease outbreaks in new locations. The survival of two model pathogens, an Oomycete, Phytophthora cinnamomi, and a filamentous fungus, Fusarium verticillioides, was studied in two different commercial potting substrates (peat and peat‐free) under glasshouse conditions in the absence of a plant host. Survival rates were analysed at 2, 7, 12 and 17 months after substrate inoculation. Fusarium verticillioides had the longest survival rate, and was still present at 17 months. In contrast, P. cinnamomi survived up to 7 months but was not recovered after 12 or 17 months. There was no significant difference in the number of colony‐forming units (CFUs) of either pathogen in the two substrates, except at 2 months, when higher numbers were recovered from peat substrates.     

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.     

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.     

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.     

Distribution of Phytophthora cinnamomi-Supprcssive Soil in Nature

Phytophthora cinnamomi-suppressivc soils were found to be widely distributed in nature. About 40 % of soil samples collected from locations with different vegetation, soil type or elevation throughout the island of Hawaii were suppressive to chlamydospore germination of Phytophthora cinnamomi. Soil samples collected from the same general areas varied greatly in degree of suppressive-ness to P. cinnamomi, ranging from conducive to strongly suppressive. Among the 155 soil samples tested, those with pH close to 4 or 8 tend to be more suppressive to P. cinnamomi than those with pH close to 6.     

Long‐term phosphite application maintains species assemblages,richness and structure of plant communities invaded by Phytophthora cinnamomi

The impact of the plant pathogen Phytophthora cinnamomi and the fungicide phosphite on species assemblages, richness, abundance and vegetation structure was quantified at three sites in Kwongkan communities in the Southwest Australian Floristic Region. Healthy and diseased vegetation treated with phosphite over 7–16 years was compared with non‐treated healthy and diseased vegetation. After site differences, disease had the greatest effect on species assemblages, species richness and richness within families. Disease significantly reduced cover in the upper and lower shrub layers and increased sedge and bare ground cover. Seventeen of 21 species assessed from the families Ericaceae, Fabaceae, Myrtaceae and Proteaceae were significantly less abundant in non‐treated diseased vegetation. In diseased habitats, phosphite treatment significantly reduced the loss of shrub cover and reduced bare ground and sedge cover. In multivariate analysis of species assemblages, phosphite‐treated diseased plots grouped more closely with healthy plots. Seven of 17 susceptible species were significantly more abundant in phosphite‐treated diseased plots compared with diseased non‐treated plots. The abundance of seven of 10 Phytophthora‐susceptible species was significantly higher along transects in phosphite‐treated vegetation. Comparison of the floristics of healthy non‐treated with healthy‐treated plots showed no significant differences in species assemblages. Of 21 species assessed, three increased in abundance and only one decreased significantly in phosphite‐treated healthy plots. In three Kwongkan communities of the SWAFR, P. cinnamomi had a profound impact on species assemblages, richness, abundance and vegetation structure. There was no evidence of adverse effects of phosphite treatment on phosphorus‐sensitive species, even after fire. Treatment with phosphite enhanced the survival of key susceptible species and mitigated disease‐mediated changes in vegetation structure. In the absence of alternative methods of control in native communities, phosphite will continue to play an important role in the protection of high priority species and communities at risk of extinction due to P. cinnamomi.     

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.     

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.     

Invasion of Euryops floribundus in degraded South African communal grassland: Unpacking the invasion relationship with elevation,soil properties,the quality and quantity of the herbaceous layer

Shrub invasion is a recent occurrence in African grassland ecosystems. This study assessed the distribution of Euryops floribundus (a native shrub species) along an elevation and invasion intensity gradients, and its relationship with soil and herbaceous vegetation in a South African grassland. Shrub density increased (p < 0.05) from the uplands (2,301 plants/ha) to the bottomlands (4,888 plants/ha). Themeda triandra (a highly palatable grass) was dominant at the uplands and sloping sites, whereas Eragrostis plana and Sporobolus africanus (poorly palatable grasses) dominated the bottomlands. Upland soils had the highest organic carbon (OC), phosphorous (P), calcium and nitrogen (N) contents, but the lowest shrub density and herbaceous biomass. Canonical correspondence analysis indicated that elevation, soil OC and N contents determine important vegetation variables along the elevation gradient. In the shrub density gradient, soil P, OC and N responded positively to E. floribundus density. Grass biomass in the moderate and heavily invaded sites was high, but the abundance of palatable grasses declined, suggesting that E. floribundus invasion may be concomitant with a decrease in rangeland condition in terms of quality forage provision.     

采用广义加法模型整合数字高程模型和遥感数据进行植被分布预测

为了采用广义加法模型整合数字高程模型和遥感数据进行植被分布的预测, 并探索耦合环境变量和遥感数据作为预测变量是否能够有效地提高植被分布预测的精度, 选择海拔、坡度、至黄河最近距离、至海岸线最近距离, 以及从SPOT5遥感影像中提取的光谱变量作为预测变量, 采用广义加法模型整合环境变量和光谱变量, 建立植被分布预测模型。研究设置3种建模情景(以环境变量作为预测变量, 以光谱变量作为预测变量, 综合使用环境变量与光谱变量作为预测变量)对黄河三角洲的优势植被类型的分布进行了预测, 并对预测结果采用偏差分析、受试者工作特征曲线和野外采样点对比等3种方法进行了验证。结果表明: (1)基于广义加法模型的植被分布预测方法具有一定的实用性, 可以较为准确地预测植被的分布; 盖度较高的植被类型预测精度较高, 盖度较低的植被类型预测精度较低, 植物群落结构的特点是出现这些差异的主要原因; 综合使用环境变量和光谱变量作为预测变量的模型, 预测精度高于单独以环境变量或者光谱变量作为预测变量的模型。(2)环境变量、光谱变量大多被选入模型, 二者均对植被分布预测有重要的作用; 同一预测变量在不同植被类型的预测模型中的贡献不同, 这与植被的光谱、环境特征差异有关; 同一预测变量在不同的建模情景下对模型的贡献不同, 环境变量与光谱变量的耦合效应可能是导致预测变量对模型的贡献出现变化的原因。     

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.     

Managing road corridors to limit fire hazard. A simulation approach in southern France

Road corridors are sources of fire ignition and fire spread in French Mediterranean areas, but little is known about the flammability of vegetation and the probability of fire ignition and spread to neighbouring forests. This study simulated fire propagation in road corridors with the help of a cellular automata (CA). We assessed the relative importance of vegetation type, fuel treatment and the spatial patterning of vegetation on the probability of fire spreading to forests. The cellular automaton simulator was implemented with different types of corridors (30 m × 40 m), on the basis of an extensive field survey of vegetation. We used data from laboratory flammability experiments to determine the probability of ignition and propagation for each of the 20 cm × 20 cm cells of the CA. The probability of a fire reaching the neighbouring forest (P_FR) indicated that certain types of road corridors represent a very high risk owing to a combination of highly flammable vegetation and high spatial connectivity. The lowest P_FR values correspond to corridors with a decreasing vegetation flammability gradient in the vicinity of the forest. A several meter-wide embankment with low-flammable and/or managed vegetation can substantially reduce P_FR. These results suggest that firewise landscaping and local vegetation management can reduce fire risk in road corridors. Each corridor type should be subjected to specific vegetation management to account for flammability, growth pattern and lifetime.     

In vitro Antagonism of Phytophthora cinnamomi and P. citricola by Isolates of Trichoderma spp. and Gliocladium virens

Three fungi, isolated from soil from which Phytophthora was not obtained, were evaluated for antagonism of Phytophthora spp. shown to cause root rot of chestnut in South Australia. Trichoderma hamatum and T. pseudokoningii appeared to inhibit P. cinnamomi by mycoparasitism. with evidence of parallel growth and coiling, and both Trichoderma spp. and Gliocladium virens grew over P. cinnamomi in vitro, preventing further growth of this pathogen. Antibiotics produced by young T. hamatum cultures and G. virens in culture filtrate experiments inhibited growth of P. cinnamomi and P. citricola. with filtrate from 4-day-old cultures of G. virens showing the greatest potential for biocontrol. All three antagonists prevented P. cinnamomi and P. citricola from causing infection symptoms on micropropagated shoots of chestnut cvs Goldsworthy and Buffalo Queen in an in vitro excised shoot bioassay for biocontrol.     

The ecological impact of Phytophthora cinnamomi in the Stirling Range National Park,Western Australia

Abstract An assessment of the impact of the soil-borne pathogenic fungus, Phytophthora cinnamomi (Oomycetes, Peronosporales), on the plant communities of the Stirling Range National Park was carried out between December 1988 and April 1989. A total of 541 plant species were collected, with the Proteaceae and the Myrtaceae the two largest families in the study region. Of the 330 species assessed for susceptibility to P. cinnamomi, 118 (36%) were recorded as having at least some individuals in a population judged to have been killed by the fungus and 33 (10%) were highly sensitive to the pathogen (more than 80% of plants in a population killed). Several families had large numbers of susceptible species, while others were apparently unaffected by the pathogen. Notably, 85% of proteaceous species assessed were rated as susceptible to P. cinnamomi. Proteaceous elements had a mean projective foliage cover of 40% in healthy plant communities, but had a mean cover of only 10% at sites that had a long history of infestation with the fungus. In contrast, some species with low levels of susceptibility to the pathogen, such as some monocotyledons, were found to be more abundant at old-infested sites than at healthy sites. Growth form may also influence susceptibility, with herbaceous perennials, annuals and geophytes assessed in this survey apparently unaffected by the fungus whereas 48% of woody perennials surveyed were susceptible. Changes in the floristic structure of plant communities may influence the composition of associated animal communities. In particular, vertebrate flower visitors may be vulnerable since 59% of the species with vertebrate-pollinated flowers were found to be susceptible to the pathogen. This research highlights the serious ecological impact of P. cinnamomi on native plant communities and suggests that significant components of the flora and associated fauna of the southwest of Western Australia are endangered by this virulent pathogen.     

Regulation of defence responses in avocado roots infected with Phytophthora cinnamomi (Rands)

Phytophthora cinnamomi occurs worldwide and has a host range in excess of 1,000 plant species. Avocados (Persea americana Mill) have been described as highly susceptible to this soil-borne pathogen. Here, the regulation of defence responses in avocado root seedlings inoculated with P. cinnamomi mycelia is described. A burst of reactive oxygen species (ROS) was observed 4 days after inoculation. The higher physiological concentration of H₂O₂ induced by P. cinnamomi on avocado roots had no effect on in vitro growth of the oomycete. Total phenols and epicathecin content showed a significant decrease, but lignin and pyocianidins exhibited no changes after inoculation. Also, increased nitric oxide (NO) production was observed 72 h after treatment. We studied the effects of one NO donor [sodium nitroprusside (SNP)], and one NO scavenger [2- to 4-carboxyphenyl-4,4,5,5-tetramethylimidazole-1-oxyl-3-oxide (CPTIO)] to determine the role of NO during root colonisation by P. cinnamomi mycelia. Pretreatment of the roots with CPTIO, but not with SNP, inhibited root colonisation suggesting an important role for NO production during the avocado–P. cinnamomi interaction. Our data suggest that although defence responses are activated in avocado roots in response to P. cinnamomi infection, these are not sufficient to avoid pathogen invasion.