Phytophthora himalsilva sp. nov. an unusually phenotypically variable species from a remote forest in Nepal

The Himalaya have received little investigation for Phytophthora species. In a remote forest in Western Nepal ten isolates of an unknown Phytophthora were recovered from the rhizosphere of Quercus, Castanopsis, Carpinus and Cupressus spp. The Phytophthora, formally named here as a P. himalsilva sp. nov., is homothallic with either amphigynous or paragynous antheridia and papillate, highly variable sporangia which may also be facultatively caducous. Based on ITS, β-tubulin, and cox I sequences Phytophthora himalsilva falls within Phytophthora Clade 2c together with Phytophthora citrophthora, Phytophthora meadii, Phytophthora colocasiae, and Phytophthora botryosa. It is suggested that Clade 2c has radiated within Asia. Molecular and sporangial characters indicate that P. himalsilva and P. citrophthora may share a recent common ancestor although they have diverged in their breeding systems. Although highly local the P. himalsilva isolates exhibited significant variation in growth rates and optimum temperatures for growth. This may reflect adaptation to different niches within a heterogeneous sub-tropical to temperate forest environment. Their cox I polymorphisms were also rather variable, including possible clustering for subsite. The occurrence of a previously unknown Phytophthora in a remote forest in Nepal highlights once again the plant health risk associated with moving rooted plants and soil between different bio-geographical regions of the world and the need for rapid pathological screening of potential risk organisms.     

Phytophthora polonica, a new species isolated from declining Alnus glutinosa stands in Poland

In a survey of Phytophthora associated with alder decline in Poland, several isolates of a homothallic Phytophthora sp., which could not be assigned to other taxa including Phytophthora alni subspecies, were consistently recovered from rhizosphere soil samples. Their morphology and pathogenicity, as well as sequence data for three nuclear regions (internal transcribed spacer rDNA, elongation factor-1alpha and beta-tubulin) and a coding mitochondrial DNA region (nadh1), were examined. The new Phytophthora species is characterized by the moderate to slow growth rate of its colony in carrot agar at 20 degrees C, high optimal (c. 30 degrees C) and maximum (c. 38 degrees C) growth temperatures, formation of catenulate, often lateral, hyphal swellings, large chlamydospores in agar media and in soil extract, persistent, ovoid to ellipsoid nonpapillate sporangia and large oogonia with paragynous and sometimes amphigynous antheridia. Phytophthora polonica was slightly pathogenic to alder twigs and not pathogenic to trunks of several tree species. In a phylogenetic analysis using either Bayesian inference or maximum likelihood methods, P. polonica falls in clade 8 'sensu Kroon et al. (2004)' of Phytophthora.     

Phytophthora austrocedrae sp. nov., a new species associated with Austrocedrus chilensis mortality in Patagonia (Argentina)

Phytophthora austrocedrae is a new species isolated from necrotic lesions of stem and roots of Austrocedrus chilensis. It is a homothallic species characterized by semipapillate sporangia, oogonia with amphigynous antheridia, and very slow growth (1–2 mm d⁻¹ on V-8 agar at 17.5 °C optimum temperature). Phylogenetic analysis of ITS rDNA sequence indicates that its closest relative is Phytophthora syringae, another species frequently isolated from soil and streams in A. chilensis forests.     

Morphological,Molecular and Pathological Characterization of Phytophthora amaranthi sp. nov. from Amaranth in Taiwan

In the spring of 2007, a serious disease on amaranth was noticed in several farms in the major amaranth production area in central Taiwan. Abundant oospores were found in the disease tissues. A species of Phytophthora was consistently isolated from disease tissues. The organism formed abundant oospores with smooth walls and with amphigynous antheridia in single culture. Sporangia were partially deciduous with short‐ to medium‐length pedicels. Morphological characteristics of this organism did not match any reported Phytophthora species, and the organism was named Phytophthora amaranthi. Pathogenicity tests and molecular characterization confirmed the identity of the organism as a new pathogen of amaranth and a new species of Phytophthora.     

Phytophthora chrysanthemi sp. nov., a new species causing root rot of chrysanthemum in Japan

A new species of Phytophthora was isolated from stem and root rot of chrysanthemum in the Gifu and Toyama prefectures of Japan. The species differs from other Phytophthora species morphologically, and is characterized by nonpapillate, noncaducous sporangia with internal proliferation, formation of both hyphal swellings and chlamydospores, homothallic nature, distinctive intercalary antheridia, and funnel-shaped oogonia. The new species can grow even at 35°C, with an optimum growth temperature of 30°C in V8 juice agar medium. In phylogenetic analyses based on five nuclear regions (LSU rDNA; genes for translation elongation factor 1α, β-tubulin, 60 S ribosomal protein L10, and heat shock protein 90), the isolates formed a monophyletic clade. Although the rDNA ITS region shows a high resolution and has proven particularly useful for the separation of Phytophthora species, it was difficult to align the sequences for phylogenetic analysis. Therefore, ITS region analysis using related species as defined by the multigene phylogeny was performed, and the topology of the resulting tree also revealed a monophyletic clade formed by the isolates of the species. The morphological characteristics and phylogenetic relationships indicate that the isolates represent a new species, Phytophthora chrysanthemi sp. nov. In pathogenicity tests, chrysanthemum plants inoculated with the isolates developed lesions on stems and roots within 3 days, and the symptoms resembled the ones originally observed. Finally, the pathogen’s identity was confirmed by re-isolation from lesions of infected plants.     

Endophytic bacterial flora in root and stem tissues of black pepper (Piper nigrum L.) genotype: isolation, identification and evaluation against Phytophthora capsici

Aim:  To isolate and identify black pepper ( Piper nigrum L) associated endophytic bacteria antagonistic to Phytophthora capsici causing foot rot disease.
Methods and Results:  Endophytic bacteria (74) were isolated, characterized and evaluated against P. capsici . Six genera belong to Pseudomonas spp (20 strains), Serratia (1 strain), Bacillus spp. (22 strains), Arthrobacter spp. (15 strains), Micrococcus spp. (7 strains), Curtobacterium sp. (1 strain) and eight unidentified strains were isolated from internal tissues of root and stem. Three isolates, IISRBP 35, IISRBP 25 and IISRBP 17 were found effective for Phytophthora suppression in multilevel screening assays which recorded over 70% disease suppression in green house trials. A species closest match (99% similarity) of IISRBP 35 was established as Pseudomonas aeruginosa ( Pseudomonas EF568931), IISRBP 25 as P. putida ( Pseudomonas EF568932), and IISRBP 17 as Bacillus megaterium ( B. megaterium EU071712) based on 16S rDNA sequencing.
Conclusion:  Black pepper associated P. aeruginosa , P. putida and B. megaterium were identified as effective antagonistic endophytes for biological control of Phytophthora foot rot in black pepper.
Significance and Impact of the Study:  This work provides the first evidence for endophytic bacterial diversity in black pepper stem and roots, with biocontrol potential against P. capsici infection.     

Evaluation of Oospore Hyperparasites for the Control of Phytophthora Crown Rot of Pepper

Nine isolates of known oospore mycoparasites comprised of six actinomycetes (Actinoplanes missouriensis, A. philippinensis, A. utahensis, Amorphosporangium auranticolor, Ampullariella regularis, Spirillospora albida) and three fungi (Acremonium sp., Humicola fuscoatra, Verticillium chlamydosporium) were tested in the greenhouse for their ability to suppress or delay the onset of crown rot of pepper caused by Phytophthora capsici. Verticillium chlamydosporium applied as a root dip increased the number of healthy plants by more than 100% when peppers were transplanted into soil artificially infested with oospores of Phytophthora capsici, but not when peppers were transplanted into soil naturally infested with P. capsici. The other mycoparasites were ineffective in the greenhouse. All the mycoparasites tested parasitized oospores of P. capsici in vitro.     

Susceptibility of Thirty Cherry Genotypes on Phytophthora cactorum, P. citrophthora, P. citricola and P. parasitica

The diseases Phytophthor a crown and root rot consist of the most important problems in cherry cultivation. In this study, the susceptibility of 30 cherry genotypes to Phytophthora cactorum , P. citrophthora, P. parasitica and P. citricola was evaluated by using excised twig assay, excised shoot method and stem inoculation method. The results showed that all cherry genotypes tested were susceptible to all Phytophthora isolates used. Phytophthora parasitica and P. citrophthora were the most aggressive species. Host specificity of the Phytophthora isolates used in this study was not found although these isolates were from different plant species. In conclusion, because none of the cherry genotype showed a level of resistance to these pathogens, caution should be taken when these genotypes are used in locations, where these diseases are endemic.     

铁皮石斛疫病及其病原菌

铁皮石斛疫病2001年发现在浙江义乌栽培田间,是由疫霉菌引起的。在田间,疫霉菌侵染茎基部,引起当年移植苗根腐、植株枯萎和死亡,但侵染2-3年植株幼嫩顶部仅引起顶枯症状。通过对病原菌形态学、交配型的观察,以及核糖体DNAITS序列分析,侵染铁皮石斛的5个分离菌株被鉴定为烟草疫霉菌Phytophthora nicotianae。致病性试验表明,铁皮石斛是烟草疫霉菌的寄主。     

Elicitin 172 from an isolate of Phytophthora nicotianae pathogenic to tomato

Elicitin 172, an acid protein with elicitor activity, has been isolated in true form from culture filtrates of Phytophthora nicotianae, the causal agent of crown and root rot of tomato (Lycopersicon esculentum). The M(r) (10,349 +/- 1) of the purified protein, determined by ES-MS, is identical to that calculated for parasiticein using the mean isotopic composition and assuming the occurrence of three disulfide bridges. The primary structure of elicitin 172, determined using also MALDI-MS experiments, shows complete identity with parasiticein, with elicitin 310 and a cloned elicitin gene from P. parasitica (= P. nicotianae), confirming conservation of the elicitin sequence within a single species. The protein induces necrosis (hypersensitive reaction) on tobacco, but no symptoms on tomato, when applied on the leaves. Tomato pretreated with elicitin 172 was affected by P. nicotianae, as well as by the phytotoxic aggregates, naturally occurring with the elicitin in the non permeated dialysis fraction of culture filtrates. Finally, the elicitin induce protection of capsicum (Capsicum annuum) and vegetable marrow (Cucurbita pepo) from P. capsici.     

Occurrence of isolates of Phytophthora colocasiae in Taiwan with homothallic behavior and its significance

Leaf blight and corm rot caused by Phytophthora colocasiae are the most devastating diseases of taro. Oospores of P. colocasiae have not been considered important in the survival in natural soil because the pathogen is heterothallic and there is essentially no chance for the presence of both A1 and A2 mating types in the same host tissue. During our recent survey of the mating type distribution of P. colocasiae in Taiwan seven homothallic isolates of Phytophthora were obtained from diseased taro leaves at Tsu Chi in central Taiwan. These organisms were identified as P. colocasiae based on morphological characteristics, ITS sequence homology and pathogenicity to taro plants. The homothallic isolates of P. colocasiae segregated into A1 and A2 types in addition to the original A1A2 type during asexual reproduction and vegetative growth. The homothallic isolate and the mixture of its A1 and A2 segregants produced abundant oospores in live tissue of taro petioles on or away from soil, indicating the possibility of oospores as a survival structure and the source of genetic variation in certain areas in nature.     

The oomycete broad-host-range pathogen Phytophthora capsici

Phytophthora capsici is a highly dynamic and destructive pathogen of vegetables. It attacks all cucurbits, pepper, tomato and eggplant, and, more recently, snap and lima beans. The disease incidence and severity have increased significantly in recent decades and the molecular resources to study this pathogen are growing and now include a reference genome. At the population level, the epidemiology varies according to the geographical location, with populations in South America dominated by clonal reproduction, and populations in the USA and South Africa composed of many unique genotypes in which sexual reproduction is common. Just as the impact of crop loss as a result of P. capsici has increased in recent decades, there has been a similar increase in the development of new tools and resources to study this devastating pathogen. Phytophthora capsici presents an attractive model for understanding broad-host-range oomycetes, the impact of sexual recombination in field populations and the basic mechanisms of Phytophthora virulence. TAXONOMY: Kingdom Chromista; Phylum Oomycota; Class Oomycetes; Order Peronosporales; Family Peronosporaceae; Genus Phytophthora; Species capsici. DISEASE SYMPTOMS: Symptoms vary considerably according to the host, plant part infected and environmental conditions. For example, in dry areas (e.g. southwestern USA and southern France), infection on tomato and bell or chilli pepper is generally on the roots and crown, and the infected plants have a distinctive black/brown lesion visible at the soil line (Fig. 1). In areas in which rainfall is more common (e.g. eastern USA), all parts of the plant are infected, including the roots, crown, foliage and fruit (Fig. 1). Root infections cause damping off in seedlings, whereas, in older plants, it is common to see stunted growth, wilting and, eventually, death. For tomatoes, it is common to see significant adventitious root growth just above an infected tap root, and the stunted plants, although severely compromised, may not die. For many cucurbit fruit, the expanding lesions produce fresh sporangia over days (or even weeks depending on the size of the fruit) and the fruit often look as if they have been dipped in white powdered confectioner's sugar (Fig. 1). Generally, hyphae do not emerge from infected plants or fruit (common with Pythium infections) and all that is visible on the surface of an infected plant is sporangia. IMPORTANCE: Phytophthora capsici presents an oomycete worst-case scenario to growers as it has a broad host range, often produces long-lived dormant sexual spores, has extensive genotypic diversity and has an explosive asexual disease cycle. It is becoming increasingly apparent that novel control strategies are needed to safeguard food production from P. capsici and other oomycetes. Considering that P. capsici is easy to grow, mate and manipulate in the laboratory and infects many plant species, this pathogen is a robust model for investigations, particularly those related to sexual reproduction, host range and virulence. USEFUL WEBSITES: Phytophthora capsici genome database: http://genome.jgi-psf.org/Phyca11/Phyca11.home.html. Molecular tools to identify Phytophthora isolates: http://phytophthora-id.org.     

BROWN ROOT ROT OF TOMATOES

Tomato roots with brown root rot showed three types of lesion: cortical rot of fine roots, 'corky root', and basal stem rot. The fungi most commonly isocated from diseased roots were: Colletotrichum atramentarium, Chaetomium spp., Cephalosporium spp., Volutella ciliata , and a grey mycelial fungus sometimes producing pycnidia ( Pyrenochaeta sp.). Other fungi isocated less frequently were: Myrothecium roridum, Petriella asymmetrica, Trichoderma airide, Phytophthora spp., Alternaria sp. and Fusarium spp. There were differences between the numbers of each species isocated from the three types of lesion in steamed and in unsteamed soils, and some seasonal variation.
Inoculation experiments with seedlings in vitro showed that Ckaetomium cochliodes and Petriella asymmetrica could infect radicles. On older plants growing in soil Colletotrichum atramentarium was the only effective pathogen. Culture filtrates from C. atramentarium, Chaetomium cochliodes , and P. asymmetrica decreased root growth when tested in vitro. Leachates from loam and a loam-manure mixture decreased the growth of tomato root-tip cultures; the effect of these leachates was altered by autoclaving.     

辣椒疫霉菌侵染模型和侵染条件定量研究

在生长箱内控制条件下分析测定了土壤温度、水分含量对辣椒疫病死苗率的影响.结果表明:土壤温度和水分状况是决定辣椒疫病菌侵染的重要因子,病菌侵染的最适土壤温度为22 ℃～28 ℃,最适土壤含水量为40%,土壤过于干燥和过饱和都不利于病菌侵染发病;辣椒疫病死苗率与土壤温度、水分含量及其互作可用数学模式描述.田间调查发现,辣椒疫病田间流行趋势可用Gompertz模型描述,发病率与初始发病率、土壤温度、水分含量以及空气温度密切相关.建立了田间辣椒疫病发病率预测模型.     

BAC-derived markers converted from RFLP linked to Phytophthora capsici resistance in pepper (Capsicum annuum L.)

Phytophthora capsici Leonian, an oomycete pathogen, is a serious problem in pepper worldwide. Its resistance in pepper is controlled by quantitative trait loci (QTL). To detect QTL associated with P. capsici resistance, a molecular linkage map was constructed using 100 F(2) individuals from a cross between Capsicum annuum 'CM334' and C. annuum 'Chilsungcho'. This linkage map consisted of 202 restriction fragment length polymorphisms (RFLPs), 6 WRKYs and 1 simple sequence repeat (SSR) covering 1482.3 cM, with an average interval marker distance of 7.09 cM. QTL mapping of Phytophthora root rot and damping-off resistance was performed in F(2:3) originated from a cross between resistant Mexican landrace C. annuum 'CM334' and susceptible Korean landrace C. annuum 'Chilsungcho' using composite interval mapping (CIM) analysis. Four QTL explained 66.3% of the total phenotypic variations for root rot resistance and three 44.9% for damping-off resistance. Of these QTL loci, two were located close to RFLP markers CDI25 on chromosome 5 (P5) and CT211A on P9. A bacterial artificial chromosome (BAC) library from C. annuum 'CM334' was screened with these two RFLP probes to obtain sequence information around the RFLP marker loci for development of PCR-based markers. CDI25 and CT211 probes identified seven and eight BAC clones, respectively. Nine positive BAC clones containing probe regions were sequenced and used for cytogenetic analysis. One single-nucleotide amplified polymorphism (SNAP) for the CDI25 locus, and two SSRs and cleaved amplified polymorphic sequence (CAPS) for CT211 were developed using sequences of the positive BAC clones. These markers will be valuable for rapid selection of genotypes and map-based cloning for resistance genes against P. capsici.     

Phylogenetic relationship of Phytophthora cryptogea Pethybr. & Laff and P. drechsleri Tucker

The phylogeny and taxonomy of Phytophthora cryptogea and Phytophthora drechsleri has long been a matter of controversy. To re-evaluate this, a worldwide collection of 117 isolates assigned to either P. cryptogea, P. drechsleri or their sister taxon, Phytophthora erythroseptica were assessed for morphological, physiological (pathological, cultural, temperature relations, mating) and molecular traits. Multiple gene phylogenetic analysis was performed on DNA sequences of nuclear (internal transcribed spacers (ITS), ß-tubulin, translation elongation factor 1α, elicitin) and mitochondrial (cytochrome c oxidase subunit I) genes. Congruence was observed between the different phylogenetic data sets and established that P. drechsleri and P. cryptogea are distinct species. Isolates of P. drechsleri form a monophyletic grouping with low levels of intraspecific diversity whereas P. cryptogea is more variable. Three distinct phylogenetic groups were noted within P. cryptogea with an intermediate group providing strong evidence for introgression of previously isolated lineages. This evidence suggests that P. cryptogea is an operational taxonomic unit and should remain a single species. Of all the morphological and physiological traits only growth rate at higher temperatures reliably discriminated isolates of P. drechsleri and P. cryptogea. As a homothallic taxon, P. erythroseptica, considered the cause of potato pink rot, is clearly different in mating behaviour from the other two species. Pathogenicity, however, was not a reliable characteristic as all isolates of the three species formed pink rot in potato tubers. The phylogenetic evidence suggests P. erythroseptica has evolved from P. cryptogea more recently than the split from the most recent common ancestor of all three species. However, more data and more isolates of authentic P. erythroseptica are needed to fully evaluate the taxonomic position of this species.