Phytophthora oleae,a new root pathogen of wild olives

Wild olive (Olea europaea subsp. europaea var. sylvestris) is an important component of Mediterranean forests and a key genetic source for olive improvement programmes. Since 2009, a severe decline caused by Phytophthora cryptogea and P. megasperma has been detected in a protected wild olive forest of high ecological value (Dehesa de Abajo, Seville, Spain). In this natural forest, sampling of roots and soil was carried out on 25 wild olives with symptoms in 2014 and 2015. Apart from the already known P. cryptogea A1 and P. megasperma, a third Phytophthora species was consistently isolated from wild olive rootlets with symptoms. These isolates conformed morphologically with the newly described species P. oleae and were confirmed by analysis of their ITS regions and cox1 sequences. Temperature–growth relationships showed a maximum growth at 19.9 °C on carrot agar medium, making it the lowest temperature Phytophthora species infecting wild olive roots. Pathogenicity was confirmed on 1-year-old healthy wild olive seedlings and was similar to the previously known pathogenic phytophthoras. As temperature requirements are quite different, the three Phytophthora species may be active against wild olive roots in different seasons. However, the prevalence of P. oleae infecting wild olives in recent years could be due to its introduction as a new invasive pathogen. The probable invasive nature of P. oleae, together with increasing rain episodes concentrated in short periods frequent in southern Spain, would allow the outbreak of infections in wild olive forests, and also put cultivated olive orchards at risk.     

疫霉菌对霜脲氰抗性遗传及对霜脲氰和甲霜灵的交互抗性

就疫霉菌对霜脲氰抗性的产生和遗传，以及疫霉菌对霜脲氰和甲霜灵的交互抗性进行了研究。结果表明，苎麻疫霉容易对霜脲氰产生抗药性突变，但抗性在连续３代单游动孢子分离过程中逐渐丧失。大雄疫霉对霜脲氰不易产生抗性突变，恶疫霉、大雄疫霉和苎麻疫霉对甲霜灵和霜脲氰不存在交互抗性     

Fungicide modes of action and resistance in downy mildews

Among oomycetes, Plasmopara viticola on grape and Phytophthora infestans on potato are agronomically the most important pathogens requiring control measures to avoid crop losses. Several chemical classes of fungicides are available with different properties in systemicity, specificity, duration of activity and risk of resistance. The major site-specific fungicides are the Quinone outside inhibitors (QoIs; e.g. azoxystrobin), phenylamides (e.g. mefenoxam), carboxylic acid amides (CAAs; e.g. dimethomorph, mandipropamid) and cyano-acetamide oximes (cymoxanil). In addition, multi-site fungicides such as mancozeb, folpet, chlorothalonil and copper formulations are important for disease control especially in mixtures or in alternation with site-specific fungicides. QoIs inhibit mitochondrial respiration, phenylamides the polymerization of r-RNA, whereas the mode of action of the other two site-specific classes is unknown but not multi-site. The use of site-specific fungicides has in many cases selected for resistant pathogen populations. QoIs are known to follow maternal, largely monogenic inheritance of resistance; they bear a high resistance risk for many but not all oomycetes. For phenylamides, inheritance of resistance is based on nuclear, probably monogenic mechanisms involving one or two semi-dominant genes; resistance risk is high for all oomycetes. The molecular mechanism of resistance to QoIs is mostly based on the G143A mutation in the cytochrome b gene; for phenylamides it is largely unknown. Resistance risk for CAA fungicides is considered as low to moderate depending on the pathogen species. Resistance to CAAs is controlled by two nuclear, recessive genes; the molecular mechanism is unknown. For QoIs and CAAs, resistance in field populations of P. viticola may gradually decline when applications are stopped.     

Effect of null mutations in the AbNIK1 gene on saprophytic and parasitic fitness of Alternaria brassicicola isolates highly resistant to dicarboximide fungicides

Alternaria brassicicola field isolates highly resistant to dicarboximide fungicides (DCF) and carrying non‐sense codons or micro‐deletions in the AbNIK1 gene encoding a group III histidine kinase were characterized at the protein level using specific antibodies. These isolates, as well as laboratory mutants expressing the same resistant phenotype, were shown to correspond to null mutants. The fitness of these isolates was compared to that of wild‐type isolates by measuring i) mycelial growth, ii) sporulation and iii) conidial germination under standard conditions. None of these factors were affected in the resistant isolates. Mycelial growth was also measured under stress conditions. Unlike suboptimal incubation temperatures or exposure to the superoxide ion generator menadione, osmotic stress generated by high sorbitol concentrations in the culture medium significantly affected the growth of AbNIK1‐null mutants compared to wild‐types. The effects of these mutations on parasitic fitness were estimated under both controlled and field conditions after inoculation of radish with individual isolates, to measure their aggressiveness, or with mixed conidial suspensions, to measure their competitive ability against wild‐type isolates. These tests on radish seedlings revealed that DCF‐resistant isolates were still aggressive. Despite this, the data obtained in the competition experiments showed that the loss of the ability to synthesize AbNIK1p in A. brassicicola DCF‐resistant mutants was linked to a severe decrease in their competitiveness.     

Identification of Phytophthora species on fruit crops in Turkey by polyacrylamide gel electrophoresis1

To identify 11 Phytophthora isolates obtained from lemon, peach and apple trees, and from strawberry plants, in the Mediterranean region of Turkey, protein, acetylesterase, peroxidase and catalose band patterns of these isolates and the same bands of 10 identified isolates of Phytophthora citrophthora, P. cinnamami and P. cactarum were compared. These 11 regional isolates were identified on the basis of the similarity of their protein and enzyme band patterns to those of identified Phytophthora isolates. It was concluded that protein band patterns could be used for identification of Phytophthora species. With enzyme band patterns, however, it may be possible to identify at species level but it is more practical to use this method only for identification of subspecies.     

Prevalence of Phytophthora species in macadamia orchards in Australia and their ability to cause stem canker

In Australia, Phytophthora cinnamomi is the only species reported as the causal agent of stem canker and root rot in macadamia. In other countries, five Phytophthora species have been reported to cause diseases in macadamia, which led us to question if more than one Phytophthora species is responsible for poor tree health in macadamia orchards in Australia. To investigate this, samples were collected from the rhizosphere, stem, and root tissues of trees with and without symptoms, nurseries, and water sources from 70 commercial macadamia orchards in Australia. Phytophthora isolates were identified based on morphological characteristics and DNA sequencing. P. cinnamomi was the most predominant and widely distributed species, and was obtained from the different types of samples including symptomless root tissues. In addition to P. cinnamomi, only P. multivora was isolated from diseased tissue (stem canker) samples. Six other Phytophthora species were obtained from the rhizosphere samples: P. pseudocryptogea, P. citrophthora, P. nicotianae, P. gondwanense, P. sojae, and a new Phytophthora taxon. Only P. cinnamomi was obtained from macadamia nursery samples, while five Phytophthora species were obtained from water sources. Of the heterothallic Phytophthora species, mating type A2 isolates were dominant in P. cinnamomi isolates, whereas only mating type A1 isolates were obtained for P. nicotianae, P. pseudocryptogea, and P. citrophthora. Pathogenicity assays revealed that P. cinnamomi and P. multivora caused significantly larger stem and leaf lesions than P. citrophthora, P. nicotianae, and P. pseudocryptogea. Phytophthora sp. and P. sojae were nonpathogenic towards leaves and stems.     

Phytophthora lateralis discovered in an old growth Chamaecyparis forest in Taiwan

The geographic origins of the invasive Phytophthora species, P. lateralis and P. ramorum are unknown. In 2008 soil samples were collected in an old growth yellow cedar (Chamaecyparis obtusa var. formosana) stand in the Ma‐kau Ecological Park in north eastern Taiwan and subjected to Phytophthora baiting procedures at 18°C. Cedar needle baits yielded isolates of a slow growing Phytophthora culture from one soil sample, together with P. cinnamomi. Phytophthora bisheria sp. nov. was obtained from another sample. The slow growing isolates conformed closely to P. lateralis in the morphology of their sporangia and chlamydospores, growth–temperature relationships, absence of gametangia and their ITS and cox II sequences. The isolates’ sporangia were partially caducous, with short pre‐formed pedicels of ca. 3–5 μm, a highly unusual feature in a non‐papillate Phytophthora. The isolates also produced multicellular stromata on cedar decoction agar. Small morphological and molecular differences were observed between the Taiwan‐isolates and Oregon‐control isolates. Taiwan may lie within the geographic centre of origin of P. lateralis. By analogy Japan may also lie within the natural range of P. lateralis; and Japan, along with Taiwan and Yunnan, could be an origin for the closely related P. ramorum.     

Present status of Phytophthora species in the Mediterranean area,especially in relation to citrus1

Of the 44 currently accepted species of Phytophthora, 18 have been reported in the Mediterranean area. The status of each is briefly reviewed. On citrus, P. citrophthora and P. nicotianae var. parasitica are mainly responsible for foot rot and gummosis, and in addition P. hibernalis, P. syringae, P. cifricola and P. cactorum for brown rot of fruits. The incidence of these species is closely linked to their temperature requirements. The use of sour orange as a resistant rootstock has long provided satisfactory control of foot rot and gummosis, while chemical treatments are effective against brown rot. However, there are indications that, with changingcultural practices, the resistance of sour orange is less well expressed, and the species is incidentally also susceptible to citrus tristeza closterovirus. Alternative rootstocks are therefore being sought.     

Phytophthora species distribution in South African citrus production regions

Several Phytophthora spp. are known to cause a range of symptoms on citrus, resulting in significant crop losses worldwide. In South Africa, Phytophthora remains a destructive citrus disease, but the species and their distribution have not been well documented. A total of 162 Phytophthora isolates was collected from 60 citrus orchards in seven provinces of South Africa (Eastern Cape, Kwazulu-Natal, Limpopo, Mpumalanga, Northern Cape, North West and Western Cape). Isolates were identified to the species level through PCR-RFLP (restriction fragment length polymorphism) analyses of the internal transcribed spacer region. The identity of a subset of the isolates was confirmed using morphological and sequence analyses. Phytophthora nicotianae was the predominant species (76 % of isolates) and occurred in 80 % of the orchards in all of the provinces, followed by P. citrophthora (22 % of isolates in 28 % of orchards). The P. citrophthora isolates were further subdivided into two previously identified subgroups, G1 and G2, with most (69 %) of the isolates belonging to the G1 subgroup. Other Phytophthora species included P. multivora in the Western Cape Province, and an unknown species in the Eastern Cape Province with high sequence similarity (98 %) to a putative new species submitted to GenBank as Phytophthora taxon Sisuluriver. Phytophthora palmivora, a known citrus pathogen, was not identified. Most of the P. nicotianae isolates (79 %) were of the A1 mating type. The P. citrophthora isolates were mostly sterile (64 %), including most of the G1 isolates (81 %). The remaining G1 isolates (19 %) belonged to the A1 mating type, whereas almost all G2 isolates belonged to the A2 mating type except for one isolate that was sterile.     

Molecular characterization of <Emphasis Type="Italic">Phytophthora porri</Emphasis> and closely related species and their pathogenicity on leek (<Emphasis Type="Italic">Allium porrum</Emphasis>)

White tip, caused by Phytophthora porri, is a destructive disease in the cultivation of European leek (Allium porrum). P. porri and closely related species such as P. brassicae, P. primulae and P. syringae belong to the phylogenetic clade 8b within the genus Phytophthora. The objectives of this study were to establish the position of P. porri and closely related species within the Phytophthora clade 8b; to study genetic variation among P. porri isolates from leek and closely related species and to test the hypothesis that host-driven speciation has occurred within this clade. AFLP analysis could clearly make a distinction between isolates of P. porri from Allium species and related Phytophthora species such as P. brassicae, P. syringae and P. primulae. DNA similarity and cluster analysis based on 353 markers demonstrated little genetic diversity within the P. porri population from Allium species although Belgian and Dutch P. porri isolates from leek could be distinguished from Japanese P. porri isolates from other Allium species and the P. porri isolate from carrot. Our results point to incipient speciation within the P. porri isolates, which could have been driven by the host plant or by geographic isolation. ITS sequence analysis confirmed the results obtained by AFLP and showed a close relationship between P. porri isolates from Allium and P. primulae and between the P. porri isolate from carrot and P. brassicae. We hypothesize that interspecific hybridization has occurred within this clade.     

Characterization of fungi (Fusarium and Rhizoctonia) and oomycetes (Phytophthora and Pythium) associated with apple orchards in South Africa

Several species of fungi and oomycetes including Fusarium, Rhizoctonia, Phytophthora and Pythium have been reported as root pathogens of apple where they contribute to a phenomenon known as apple replant disease. In South Africa, little is known about specific species in these genera and their pathogenicity toward apple. Therefore, these aspects were investigated along with the development and optimization of qPCR tests for detection and quantification of the most virulent oomycete species. In eight investigated orchards, the oomycete Phythophthora cactorum was widely distributed, while nine Pythium species were differentially distributed among the orchards. Pythium irregulare was the most widely distributed and the most virulent species along with P. sylvaticum, P. vexans and Ph. cactorum. Seven binucleate Rhizoctonia anastomosis groups (AGs) were also differentially distributed among the orchards, with the majority appearing to be non-pathogenic while certain AG-I and AG-F isolates exhibited low virulence on apple. In the genus Fusarium, F. oxysporum was widely distributed, but isolates were non-pathogenic. Fusarium solani and F. avenaceum were less frequently encountered, with only some isolates having low virulence. qPCR data obtained from seedling roots inoculated with the most virulent Pythium species (P. irregulare, P. sylvaticum and P. vexans) and the genus Phytophthora were not always reproducible between trials, or isolates of the same species. In general, seedling growth inhibition was associated with the presence of a low amount of pathogen DNA (±40 fg μl⁻¹ to 2 pg μl⁻¹) in roots. Pythium irregulare, although having the lowest DNA concentrations in roots, was the only species for which a significant negative correlation was found between seedling weight and pathogen DNA concentration.     

Targets for pathology research in protected crops

Due to the need to diminish the amount of pesticides used, alternative ways of controlling fungal diseases have to be developed. Foliar diseases have been managed mainly by chemicals, but research has been started to develop integrated control programmes. Data from studies on epidemiology, in combination with nutritional and climatic management and use of partial resistant cultivars will lead to a reduced use of chemicals. Models used are: Botrytis cinerea in gerbera and roses, and powdery mildew in roses and cucumbers. Research has been intensified on biological control of these pathogens. In closed systems, with recirculation of the nutrient solution, soil-borne fungi can cause serious problems. Not only do diseases known from traditional cultural systems, like fusarium wilt of carnation, fusarium crown and root rot of tomato and Phytophthora and Pythium spp. in several crops occur, but also new problems like a new Phytophthora sp. and Gnomonia sp. in roses and a Cylindrocladium sp. in Spathiphyllum spp. To prevent introduction of pathogens in recirculation systems, emphasis is put on developing an integrated disease management programme by using disease-free planting material, disease-free irrigation water, strict hygienic measures, resistant cultivars, methods of disinfecting the nutrient solution and biological control. Research over the last 5–6 years has revealed good prospects for biological control, especially in closed systems with a limited amount of substrate. Wilt in carnation, caused by Fusarium oxysporum f.sp. dianthi can be prevented effectively by adding a non-pathogenic isolate of F. oxysporum and/or Pseudomonas spp. There are indications that non-pathogenic isolates of F. oxysporum are also effective against wilt disease in other crops. An isolate of Trichoderma harzianum appears to be very effective against fusarium crown and root rot of tomato. More applications of this, and other, biocontrol agents seem possible. However, translation of the results of research to practical application and registration is still very difficult.     

Characterisation and detection of Pythium and Phytophthora species associated with grapevines in South Africa

Replant and decline diseases of grapevines not only cause quantitative and qualitative yield losses, but also result in extra costs when vineyards have to be replanted. This study investigated the role of Pythium and Phytophthora in the decline syndrome in South Africa by determining (1) the species associated with nursery and established vines, and (2) pathogenicity of Ph. sp. niederhauserii and P. vexans relative to known grapevine pathogens. Quantitative real-time PCR (qPCR) assays were also developed for detection of the most prevalent oomycete groups. In total, 26 Pythium and three Phytophthora species were identified from grapevine nurseries and established vineyards. The most common infections in sampled nursery vines were caused by P. vexans (16.7%), followed by P. ultimum var. ultimum (15.0%) and P. irregulare (11.7%). In established vineyards, P. irregulare (18.0%) and P. vexans (6.2%) were also among the three most prevalent species, along with P. heterothallicum (7.3%). Three Phytophthora species were also identified from the sampled established vines, of which Ph. cinnamomi (5.1%) was predominant, followed by Ph. sp. niederhauserii (1.1%). In established vineyards a higher incidence and more diverse species composition was observed in spring and winter, than in summer. Pathogenicity studies showed that some Ph. sp. niederhauserii and P. vexans isolates were as aggressive as the known grapevine pathogens Ph. cinnamomi and P. irregulare. Sensitive qPCR assays were developed for the detection of P. ultimum var. ultimum, P. irregulare, P. vexans and the genus Phytophthora. These assays will be invaluable in limiting pathogen dispersal through screening of nursery material. This is especially important since pathogenic species were also isolated from healthy looking vines in nurseries.     

Morphological and molecular characterization of Phytophthora species associated with root and crown rot of pomegranate in Iran

The purpose of this research was to identify the pathogens causing root and crown rot in major pomegranate-growing areas of Iran. Infected tissue samples were collected from trees with symptoms from 49 pomegranate orchards in four provinces of Iran: Fars, Markazi, Isfahan, and Kohgiluyeh va Boyer-Ahmad. In total, 23 Phytophthora spp. isolates were obtained, which were identified as P. cryptogea species complex (12 isolates) and P. cinnamomi (11 isolates) based on morphological characters. Molecular confirmation of identification was performed by inference of phylogeny of ITS-rDNA regions, β-tubulin gene, and the mitochondrial gene cytochrome c oxidase subunit 1. The results of our phylogenetic analysis confirmed the morphological identification of P. cinnamomi isolates and placed them in Clade 7c of Phytophthora. In addition, the P. cryptogea species complex isolates, despite morphological similarities, were in fact four distinct species including P. cryptogea sensu stricto (two isolates), P. pseudocryptogea (one isolate), P. sp. kelmania (one isolate), and P. erythroseptica (six isolates). This is the first report of pomegranate root and crown rot caused by P. cinnamomi and P. cryptogea species complex.     

Phytophthora boehmeriae boll rot: a new threat to cotton cultivation in the mediterranean region

A boll rot of cotton (Gossypium hirsutum L.) was observed for the first time in Greece in August 1993 in Larissa and Volos counties, and in August and September 1995 in Trikala and Phthiotis counties. Fungi of the genusPhytophthora were isolated from diseased plants. Morphological characteristics of the pathogen were recorded on mounts made directly from the infected tissues or after growth of the isolated fungus on corn meal agar or sterile distilled water. Colony morphology, growth rates, features of asexual and sexual structures and maximum growth temperatures were examined. APhytophthora species new to Europe,Phytophthora boehmeriae Sawada, attacking cotton bolls, was identified. The pathogenicity of the isolates was confirmed by artificial inoculations of detached cotton bolls. Analysis of α-esterase isozymes revealed unique banding patterns for isolates ofP. boehmeriae compared with those ofP. cactorum andP. parasitica, which arePhytophthora species with similar morphology.     

Antimicrobials in Phytophthora isolation media and the growth of Phytophthora species

Many recently described Phytophthora species detected using high-throughput sequencing have never been isolated into culture. NARH is a commonly used isolation medium containing cornmeal agar with n ystatin 22.72 ppm, a mpicillin 100 ppm, r ifampicin 10 ppm, and h ymexazol 50 ppm. We investigated whether the antimicrobial compounds in this medium selectively inhibit growth of some Phytophthora species. Growth of 10 Phytophthora species from 10 Phytophthora clades was tested in NARH medium with antimicrobials in a range of concentrations, singly and in combination: nystatin 12.5–100 ppm, ampicillin 62.5–500 ppm, rifampicin 5–40 ppm, hymexazol 12.5–100 ppm and in addition, chloramphenicol 5–40 ppm. Two combinations of antimicrobials that supported good growth of the greatest number of species were selected: (a) nystatin 12.5 ppm, ampicillin 250 ppm, rifampicin 10 ppm, and hymexazol 12.5 ppm, and (b) these compounds with the addition of chloramphenicol 5 ppm. Subsequently the growth of 47 Phytophthora species from all clades was tested in media with these two combinations and standard NARH, both in aseptic culture and in plates with leaf baits infected from soils known to be infested with Phytophthora. Although growth of some Phytophthora species was better with the new combinations of antimicrobials in aseptic culture, suppression of competing microorganisms was best in standard NARH. Sensitivity to the NARH antimicrobials is not the reason for the difficulty or failure of isolating many species detected using high-throughput sequencing. NARH is a robust and appropriate medium for isolation of Phytophthora species from all clades.     

Relative parasitic fitness of isolates of Pyricularia oryzae Cav. with different sensitivities to fungicides

The relative parasitic fitness of isolates of Pyricularia oryzae with different sensitivities to isoprothiolane (di-isopropyl 1,3-dithiolan-2-ylidenemalonate) and IBP (S-benzyl O, O-di-isopropyl phosphorothioate) was studied in the absence of fungicides. Four field isolates (S-1,S-2,MR-1 and MR-2) and two in-vitro mutants (Rvt-1 and Rvt-2) were used for disease epidemics. S-l and S-2 were wild types; MR-1 and MR-2 were sensitive to isoprothiolane and moderately resistant to IBP, and Rvt-1 and Rvt-2 were resistant to both fungicides. Twelve epidemics were made by inoculating rice seedlings with mixed conidial suspensions of two isolates or mutants. The value of relative parasitic fitness (W = 0-1.0) was calculated for each isolate and epidemic. S-l and S-2 were stronger (W = 1.0) than MR-2, Rvt-1 and Rvt-2; but weaker (W = 0.75, 0.73, respectively) than MR-1. MR-1 was strongest among all isolates and mutants used. MR-2 was slightly weaker (W = 0.9-1.0) than S-l and S-2, but stronger than Rvt-1 and Rvt-2. Rvt-1 and Rvt-2 had smaller values of W, ranging from 0.25-0.58, in the epidemics with each field isolate. These results suggest that the proportions of in-vitro mutants do not increase unless intensive selection pressure is given, and would be expected to decrease rapidly after the selection pressure is removed. Isolates moderately resistant to IBP, such as MR-1 and MR-2, however, had high values of W, suggesting that they would increase or would not decrease rapidly in the absence of selection pressure. These results may well explain why isolates highly resistant to isoprothiolane and IBP have seldom been found, and why a large number of isolates moderately resistant to IBP but sensitive to isoprothiolane have been observed in the field.     

番茄灰霉病菌对咯菌腈的抗性诱变及抗药突变体的生物学特性

为评估番茄灰霉病菌Botrytis cinerea对咯菌腈的抗性风险,就室内经紫外照射获得抗药突变体的方法及抗性突变体的生物学性状进行了研究。结果表明:番茄灰霉病菌分生孢子的紫外照射亚致死时间为90~120 s;经亚致死时间紫外照射后,4个亲本菌株中有2个菌株共产生了6个抗咯菌腈的突变体,其EC50值是亲本菌株的310倍以上,抗性突变频率为3.13×10-7;经紫外照射诱变获得的所有抗性突变体在菌丝生长速率、产孢量、产菌核能力及其在番茄果实上的致病性方面均比其亲本菌株明显降低。相关分析显示,所得抗咯菌腈突变体对氟啶胺、啶菌唑、啶酰菌胺和嘧霉胺无交互抗性。表明番茄灰霉病菌对咯菌腈的抗药性风险较低。     

Specific and Sensitive Detection of Phytophthora nicotianae By Simple and Nested-PCR

Phytophthora nicotianae Breda de Haan is one of the most important soil-borne plant pathogens. The identification of this pathogen based on morphological or physiological characters is time-consuming and labour-intensive and requires comprehensive knowledge of fungi. Molecular analysis of the internal transcribed spacer (ITS) regions of rDNA is a novel and very effective method of species determination. Based on this concept, conventional and single closed tube nested-PCRs were developed for the specific and sensitive detection of P. nicotianae. Two new specific primers, designed from the spacer regions ITS1 and ITS2, internal to the nucleotide sequence flanked by universal primers ITS4 and ITS6, were used. To evaluate the specificity of the method, 36 morphologically characterized isolates were tested. A positive reaction, characterized by an amplification product of 737 bp, was shown by all P. nicotianae isolates and two P. nicotianae/cactorum hybrids. No amplification product was observed when other Phytophthora species and genera were assayed. The sensitivity of this method was analysed by serial dilutions of a defined amount of fungal DNA in a healthy root extract. Nested-PCR was at least 1000 times more sensitive than conventional PCR. In addition, samples from different infection sites, origins and crops, samples from nutrient solution, water and the rockwool used in hydroponic cultures, were analysed to validate this method.     

Control of Phytophthora species in plant stock for habitat restoration through best management practices

Emergent plant pathogens represent one of the most significant threats to biodiversity, and exotic Phytophthora species have recently emerged as a serious problem in restored habitats in California and in nurseries producing the plant stock. It is hypothesized that ‘best management practices’ prescribed through a Phytophthora Prevention Programme (PPP) could be useful in minimizing phytophthora disease incidence. To understand the magnitude of the problem and the efficacy of the PPP, plants in restoration nurseries were evaluated for (i) the Phytophthora species assemblage present in the absence of the PPP, and (ii) the effectiveness of the PPP to reduce them. Sampling included 203 plants grown in the absence of the PPP, and 294 grown implementing the PPP. Only samples collected in the absence of the PPP were Phytophthora-positive, and cumulatively yielded 55 isolates from 13 different taxa, including 1 putative interspecific hybrid genotype. There were 21 novel Phytophthora–plant species combinations. The most common Phytophthora species was P. cactorum. Four plant species had the highest disease incidence, namely: Diplacus aurantiacus (50 ± 11.2%), Heteromeles arbutifolia (33 ± 9.6%), Ceanothus thyrsiflorus (30 ± 8.4%), and Frangula californica (30 ± 8.4%). Disease incidence in nurseries after the implementation of the PPP dropped to zero (P < 0.001), and was unaffected to any significant degree by nursery differences, or plant species tested. This study identifies a large number of novel ‘plant species × Phytophthora species’ combinations, and provides for the first time strong evidence that the PPP significantly reduced Phytophthora in plant stock for habitat restoration.