Phosphorous acid treatments control Phytophthora diseases in Australia1

Treatments with a partially neutralized formulation of phosphorous acid containing potassium phosphite were assessed for control of Phytophthora diseases in subtropical and temperate crops in Australia. In Queensland, trunk injections of phosphite (10% solution) controlled severe root rot (Phytophthora cinnamomi) of avocado trees and resulted in the recovery of trees. Single pre-harvest sprays (2.5 kg ha^-1) of phosphite controlled root and heart rot (P. cinnamomi) of pineapples. Foliar sprays of phosphite (64 g per tree) controlled root rot (P. nicotianae var. parasitica) and trunk canker (P. citrophthora) of mandarin trees. In Victoria, a foliar spray of phosphite (300 g ha^-1) reduced root rot (P. clandestina) of subterranean clover and increased dry matter by 1.96 to 5.11 t ha^-1. Trunk injections of phosphite (10% solution) controlled trunk rot (P. cactorum) of peach trees and foliar sprays (10 kg ha^-1) reduced severity of root rot (P. nicotianae var. nicotianae) of tomatoes.     

Interaction between root rot basidiomycetes and Phytophthora species on pedunculate oak

Forest declines are usually complex multifactorial phenomena that involve interactions between different factors. The possible interaction between different types of mycelial pathogens was investigated through artificial inoculation of oak seedlings, involving two root rot basidiomycetes, Collybia fusipes and Armillaria mellea, and two Phytophthora species, P. cinnamomi and P. cambivora. These pathogens were inoculated onto young Quercus robur saplings in greenhouse conditions, either alone or combining a root rot basidiomycete with a Phytophthora species. Three out of the four Phytophthora spp.*root rot basidiomycete combinations tested resulted in significantly greater damage to the oak host than the sum of the damages induced by the individual pathogens. This positive interaction could be significant in oak decline syndrome.     

The effect of three irrigation practices on phytophthora crown and root rot of apple trees under field conditions

The effect of microjet, drip, and two durations of sprinkler irrigation systems on phytophthora crown and root rot of apple trees was examined under field conditions. This eight year study indicates that crown and root rot caused byPhytophthora cactorum was most severe where young MM. 106 rootstock trees were watered by microjet irrigation for 2.3 h each day. There was no difference in infection byP. cactorum when trees were irrigated either by drip or sprinkler irrigation systems. The MM.106 apple rootstock trees watered by drip irrigation for 2.6 h each day were least affected by phytophthora crown and root rot.     

Monitoring of <Emphasis Type="Italic">Phytophthora</Emphasis> species on fruit trees in Bulgaria

Disease on fruit trees in Bulgaria caused by Phytopthora cactorum and P. citrophthora was found in the period 1998–1999. Leaves of some trees become reddish during July, and later in the season fall off. Infected trees die during the same season, or the next season. Observations on symptom development and spread of Phytophthora root and crown rot of fruit trees was undertaken from 1999 to 2009. Disease incidence is between 2% and 14% in some gardens and nurseries. The disease was registered in the regions of Plovdiv, Kjustendil, Sliven, Yambol, Karnobat, Bourgas and Svishtov. Samples from infected plant tissues were taken and isolations were done on selective PARP media, or by applying a baiting bioassay. Based on morphological and cultural characteristics and temperature requirements the following Phytophthora species have been identified: Phytophthora cactorum, P. citrophthora, P. drechsleri, P. cryptogea, hybrid and Pythium. Pathogenicity of the isolates was tested on green apple fruits or one-year-old apple rootstocks. Laboratory studies of the effect of temperature on mycelia growth showed that most isolates can grow from 5° up to 30°C, with an optimum from 18° to 25°C. Only three strains grew at 35–36°C, two developed slowly, one grew well. The optimal pH for mycelia development was tested. Aiming at control of disease, in vivo pot trials have been carried out for studying resistance of rootstocks to P. cactorum. At the end of the growing season a good level of resistance has been shown in the rootstocks M29C, Gizela 6, and MAXMA 14.     

Effect of nitrogen form and application method on incidence and severity ofPhytophthora crown and root rot of apple trees

The effect of ammonium nitrate broadcast as a soil or through irrigation, urea applied as a foliar spray, and monoammonium phosphate applied as a planting hole treatment on the incidence ofPhytophthora crown and root rot of apple trees was determined under orchard conditions in the Okanagan Valley of British Columbia, Canada. Results from the eight year study showed that ammonium nitrate applied as a single dose in spring at 240 g tree^–1 year^–1, as a split dose at 120 g tree^–1 each in spring and early autumn, and in irrigation water (fertigation) at 7.5 g tree^–1 wk^–1 for 10 wk year^–1 significantly increasedPhytophthora crown and root rot of Macspur on MM106 rootstock. There was no significant difference inP. cactorum infection between the unfertilized control and treatments with urea applied as a foliar spray at 1.0 kg 100 l^–1 of water in spring and early autumn, and monoammonium phosphate applied as a planting hole treatment at 1 g l^–1 of soil at planting time.     

Preliminary evaluation of nine fungicides for control ofPhytophthora cactorum andP. citrophthora associated with crown rot in peach trees

Excised twig assay and excised stem inoculation were used to evaluate nine fungicides (metalaxyl, fosetyl-Al, copper hydroxide, copper sulfate, copper oxychloride, captan, quintozene, propamocarb and chlorothalonil) againstPhytophthora cactorum andP. citrophthora associated with crown rot in peach trees. Segments were soaked in fungicide solutions at different concentrations and then inserted vertically intoP. cactorum orP. citrophthora cultures growing on cornmeal agar plus antibiotics, or inoculated by inserting a mycelium-bearing agar plug directly into the cambium. Following incubation, the bark was scraped off and length of necrosis was measured. Metalaxyl was the only fungicide that inhibited canker development on segments at the manufacturer-recommended concentration. Fosetyl-Al, captan, copper hydroxide and copper sulfate inhibited canker development at 3, 4, 4 and 8 gl^-1, respectively. The other fungicides did not affect canker length significantly compared with non-treated twigs, with the exception of propamocarb, which reduced the development ofP. cactorum on excised stems. The tested methods enabled rapid and effective evaluation of a large number of chemicals to prevent crown rot diseases caused byPhytophthora in the laboratory. http://www.phytoparasitica.org posting Dec. 5, 2001.     

安徽省大豆疫霉根腐病菌的鉴定及rDNA-ITS序列分析

为明确安徽省夏大豆疫霉根腐病的病原菌种类,对采集自涡阳、怀远、固镇3个县的夏大豆病株及土样分离纯化后获得28株菌株,选取6株代表性菌株,通过形态学观察及核糖体DNA-ITS序列分析对其进行鉴定,并测定了其致病型。结果表明,6株菌株在利马豆培养基上菌落白色,质地均匀;菌丝无隔,致密,具近直角分枝;在10%V8C培养液中,游动孢子囊顶生,不脱落,卵形至椭圆形,无明显乳突,有内层出现象,长宽比大于1.6∶1;同宗配合,在利马豆培养基上单株培养产生大量卵孢子,藏卵器球形,雄器大多侧生;接种合丰35大豆品种后出现典型的大豆疫霉根腐病症状。r DNA-ITS序列分析表明,6株菌株与Gen Bank中大豆疫霉Phytophthora sojae的ITS序列同源性高达100%;菌株GY4、GY8、HY11、HY16、GZ10、GZ21的毒力公式分别为1b,2,3a,3b,4,5,6,7;1b,1d,3a,3b;1d,3a,3b,3c,4,5,6,7;2,3c,4,5,6,7;1b,3a,3c,5,8;3a,3b,5,6,7,8;属于6个不同的致病型。研究表明,这6株菌株均为大豆疫霉。     

Real-time detection of <Emphasis Type="Italic">Phytophthora nicotianae</Emphasis> and <Emphasis Type="Italic">P. citrophthora</Emphasis>in citrus roots and soil

Two primers, specific for Phytophthora nicotianae (Pn6) and P. citrophthora (Pc2B), were modified to obtain Scorpion primers for real-time identification and detection of both pathogens in citrus nursery soils and roots. Multiplex PCR with dual-labelled fluorogenic probes allowed concurrent identification of both species ofPhytophthora among 150 fungal isolates, including 14 species of Phytophthora. Using P. nicotianaespecific primers a delayed and lower fluorescence increase was also obtained from P. cactorumDNA. However, in separate real-time amplifications, the aspecific increase of fluorescence from P. cactorum was avoided by increasing the annealing temperature. In multiplex PCR, with a series of 10-fold DNA dilutions, the detection limit was 10 pg l^-1 for P. nicotianaeand 100 pg l^–1 for P. citrophthora, whereas in separate reaction DNA up to 1 pg l^-1 was detected for both pathogens.Simple and rapid procedures for direct DNA extraction from soil and roots were utilised to yield DNA whose purity and quality was suitable for PCR assays. By combining these protocols with a double amplification (nested Scorpion-PCR) using primers Ph2-ITS4 amplifying DNA from the main Phytophthora species (first round) and PnB5-Pn6 Scorpion and Pc2B Scorpion-Pc7 (second round), it was possible to achieve real-time detection of P. nicotianaeand P. citrophthora from roots and soil. The degree of sensitivity was similar to that of traditional detection methods based on the use of selective media. The analyses of artificially and naturally infested soil showed a high and significant correlation between the concentration of pathogen propagules and the real-time PCR cycle threshold.     

Testing variability in pathogenicity ofPhytophthora cactorum, P. citrophthora andP. syringae to apple, pear, peach, cherry and plum rootstocks

The relative virulence ofPhytophthora cactorum andP. syringae originating from almond trees, and ofP. citrophthora originating from citrus, to apple, pear, peach, cherry and plum rootstocks, was studiedin vivo andin vitro. Results of the different experiments were in good agreement. All testedPhytophthora isolates showed little virulence to pear rootstocks-causing only minor crown rot symptoms - and no virulence at all to apple rootstocks. In contrast, they were highly virulent to stone fruit rootstocks, causing crown rot disease. The non-pathogenicity of these isolates to pome rootstocks could be interpreted as strict host specificity.     

Effects of six herbicides on the growth of Phytophthora cactorum and a bacterial antagonist

Six herbicides, incorporated in corn meal agar, were tested for their effects on the growth of Phytophthora cactorum, the growth of a bacterial antagonist B8 (Erwinia sp.), and the antagonistic effect of B8 on P. cactorum Oryzalin and glyphosate significantly reduced the growth of P. cactorum. Neither the growth of B8, nor its antagonistic effect on P. cactorum, was affected by any of the herbicides tested. This suggests that herbicides may not be a limiting factor on the development of bacterial antagonists on a biological control method for the control of crown rot of apple trees.     

Phytophthora crown rot of almonds in Australia 1

Phytophthora crown rot, caused primarily by Phytophthora cambivora, has caused considerable losses in almond orchards in the Willunga and northern plains areas approximately 50 km north and south of Adelaide (AU), respectively. Other Phytophthora species including P. citrophthora, P. cryptogea, P. megasperma and P. syringae have also been associated with diseased trees. Tree losses have been associated with P. cactorum isolated from trunk cankers on scaffold limbs on trees in the Riverland almond-growing area approximately 350 km north-east of Adelaide. The Al mating type of P. cambivora was the most pathogenic to almond seedlings. Cvs Mission and Chellaston, which are commonly used as rootstocks, were highly susceptible to crown rot whereas peach cv. Nemaguard was resistant. An excised twig assay has been developed to screen micropropagated shoots for resistance to P. cambivora. Metalaxyl applied at 5 g active substance per tree in spring, winter and autumn in a shallow trench close to the trunk prevented the development of trunk cankers. Foliar sprays of phosphonate (H₃PO₃) at 2 g active substance per litre in autumn and spring also inhibited canker development.     

Effects of inoculum density,plant age and temperature on disease severity caused by pythiaceous fungi on several plants

Alfalfa, maize, sorghum and sugarbeet plants were inoculated with zoospores ofPhytophthora andPythium species in order to assess the effects of inoculum density, plant age and temperature on disease severity. Seedlings were grown axenically in test tubes and inoculated with zoospore suspensions. Disease severity was assessed by measuring the root growth and discoloration of treated and control seedlings. The incremental root length of all plants decreased and root discoloration increased as inoculum concentration of the pathogen increased. Changes were more intensive among low levels of zoospore concentrations and no significant differences in disease severity were found for inoculum densities higher than 10⁴ zoospores ml^-1. Disease severity was negatively related to plant age. Disease development on sugarbeet seedlings infected withPythium andPhytophthora species was affected by temperature, but the pattern of response was determined by the pathogen’s temperature preferences. The incremental root length decreased as temperature increased up to 25°C. The effect ofPythium dissimile andPhytophthora cactorum on root length was significantly lower at 35°C than at 25°C, whereasPythium aphanidermatum andPhytophthora nicotianae caused significant damage to roots even at 35°C. http://www.phytoparasitica.org posting Dec. 3, 2001.     

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.     

Evaluation of benomyl and thiophanate-methyl for the control of Verticillium wilt of strawberry in the Netherlands

In pot experiments the fungicides benomyl and thiophanate-methyl controlledVerticillium wilt of strawberry when applied as a soil drench after planting. Both compounds were ineffective as foliar sprays and as root dips prior to planting. Soil drenches applied to commercially grown runner plants in the waiting field (August) and to the same plants in the greenhouse (December or January) increased the yield. On infested ground, a soil drench with thiophanate-methyl promoted the occurrence of crown rot caused byPhytophthora cactorum.     

Early survival of Quercus ilex subspecies from different populations after infections and co‐infections by multiple Phytophthora species

Forests in Europe are threatened by increased diversity of Phytophthora species, but effects on trees of simultaneous infections by Phytophthora and ecological consequences of their coexistence are unknown. This study explored variation in early survival of Quercus ilex to Phytophthora infections and assessed interactions between Phytophthora species when trees were co‐infected. Three Phytophthora species (P. cinnamomi, P. gonapodyides and P. quercina), seeds from 16 populations of Q. ilex (ballota and ilex subspecies) and two infection times were used as sources of variation in two experiments. The influence of Phytophthora species, Q. ilex subspecies and populations on plant germination and survival were analysed using generalized linear mixed models and survival analysis techniques. Germination rates were not influenced by Phytophthora spp. (P = 0.194) but by the subspecies and populations of Q. ilex (P < 0.001). In Phytophthora‐infested soils, Q. ilex subsp. ilex germinated at higher rates than Q. ilex subsp. ballota. Plant survival was strongly influenced by Phytophthora species (P < 0.001), not by the subspecies and populations of Q. ilex. Seedling mortality was reduced and delayed if a less virulent Phytophthora species infected plants prior to infection by a more virulent Phytophthora species. The results help to explain oak decline syndrome and the lack of natural and artificial regeneration of Q. ilex forests. Lack of interspecific variability of early survival to Phytophthora spp. discourages direct sowing for artificial reforestation programmes. Large, thick seeds, giving plants rapid growth, are advantageous traits when soils are infested with Phytophthora spp.     

Soil receptivity toFusarium solani f. sp.pisi and biological control of root rot of pea

Potential antagonists ofFusarium solani f. sp.pisi (Fsp) were selected from soil samples with varying degrees of receptivity to this pathogen. They were tested against Fsp isolate 48 (Fs48), in increasingly complex systems. Most species testedin vitro were able to antagonize Fs48. No relation could be establishedin vitro between the receptivity of the soil from which an isolate originated and its antagonism to Fs48. In soils naturally infested with pea root rot pathogens, which were stored humid at 4°C for a period longer than a year, various isolates ofFusarium, Gliocladium andPenicillium spp. were able to reduce root rot. After sterilization of these soils, onlyGliocladium roseum isolates, added at 10⁵ conidia g^–1 dry soil, significantly reduced disease severity and prevented root weight losses caused by Fs48 at 10⁴ conidia g^–1 dry soil. In soils in which the biota were activated by growing peas before the assays, doses of 10⁶ and 10⁷ ofG. roseum were required to reduce root rot. In these soils, the antagonistic effects of fluorescent pseudomonad strains from soil of low receptivity to Fsp were variable. Some strains of fluorescent pseudomonads, from soil moderately receptive to Fsp and from highly infested soils, were also able to reduce root rot. Disease suppression by pseudomonad strains was more evident in the absence than in the presence ofAphanomyces euteiches in the root rot pathogen complex. The role of receptiveness of the soil with regard to potential antagonists is discussed.     

Detection and Identification of <Emphasis Type="Italic">Phytophthora</Emphasis> Species in Southern Italy by RFLP and Sequence Analysis of PCR-amplified Nuclear Ribosomal DNA

In four neighbouring regions of southern Italy, Basilicata, Campania, Apulia and Calabria, pepper and zucchini plants showing Phytophthora blight symptoms, tomato plants with either late blight or buckeye rot symptoms, plants of strawberry showing crown rot symptoms and declining clementine trees with root and fruit rot were examined for Phytophthora infections by means of polymerase chain reaction (PCR) assays, using primers directed to nuclear ribosomal DNA (rDNA) repeat sequences. All diseased plants and trees examined tested positive. The detected fungal-like organisms were differentiated and characterized on the basis of primer specificity as well as through extensive restriction fragment length polymorphism (RFLP) and sequence analysis of PCR-amplified rDNA. Phytophthora capsici was identified in diseased pepper and zucchini plants, P. infestans was identified in tomato with late blight symptoms whereas buckeye rot-affected tomatoes and diseased strawberry plants proved to be infected by P. nicotianae and P. cactorum, respectively. Declining clementine trees were infected with P. citrophthora and P. nicotianae in about the same proportion. Also, thirty-one pure culture-maintained isolates of Phytophthora which had previously been identified in southern Italy by traditional methods but were never examined molecularly, were examined by RFLP and sequence analysis of PCR-amplified nuclear rDNA. Among these, an isolate from gerbera which had previously been identified by traditional methods only at genus level, was assigned to P. tentaculata. For the remaining pure culture-maintained isolates examined, the molecular identification data obtained corresponded with those delineated by traditional methods. Most of the diseases examined were already known to occur in southern Italy but the pathogens were molecularly detected and fully characterized at nuclear rDNA repeat level only from other geographic areas, very often outside Italy. A new disease to southern Italy was the Phytophthora blight of zucchini. This is also the first report on the presence and molecular identification of P. tentaculata from Italy.     

Variable response of open-pollinated seedling progeny of avocado to Phytophthora root rot

Phytophthora root rot, caused byPhytophthora cinnamomi Rands, is the most important disease of avocado (Persea americana Miller). In an attempt to identify root rot-resistant rootstocks that could ultimately be used under conditions in southern Florida, we screened open-pollinated progeny of avocado from the National Germplasm Repository in Miami. From 1996 to 1998, a total of 2,355 seedlings from 51 accessions were examined in potting mix artificially infested withP. cinnamomi. Most seedlings developed severe root rot, but tolerance was observed in some families (i.e., progeny of certain accessions). Although the most susceptible families developed mean disease ratings of up to 97% root necrosis, mean ratings for the most tolerant families were less than 60%. There was also a strong relationship between the racial background of the female parent and the tolerance of seedlings. Seedlings of the West Indian race and hybrids between it and the Guatemalan race were significantly more tolerant than those from other parents (P< 0.05). Individuals in several families developed < 50% root necrosis, the arbitrary standard of tolerance in this study. Twelve families accounted for 82% (188 of 229) of the tolerant seedlings, and only two of these did not have a West Indian or Guatemalan × West Indian pedigree. Broad-sense heritability for PRR tolerance was 0.45. This is the first report on the inheritance of PRR tolerance in avocado and on the influence of genotype and racial pedigree under controlled conditions.     

Two Phytophthora species causing decline of wild olive (Olea europaea subsp. europaea var. sylvestris)

Since 2009, a severe decline leading to mortality has been observed affecting nearly 5 ha of a wild olive woodland of high ecological value in Seville, southern Spain. Phytophthora cryptogea and P. megasperma were consistently isolated from roots and rhizosphere of trees with symptoms sampled in 2009, 2011 and 2013. The isolates were identified on the basis of colony and reproductive structure morphology as well as temperature–growth relationships, and identification was further corroborated by their ITS and β‐tubulin sequences. Koch's postulates were demonstrated for both species on 1‐year‐old wild olives. Pathogenicity tests showed that both Phytophthora spp. are highly aggressive pathogens, although temperature–growth requirements for each species were distinct. As a consequence, the two species may be active in different seasons and their epidemiology may be differently influenced by global climate change, and they may show their active periods in different climatic scenarios. The climate change models for the Mediterranean Basin forecast a global temperature increase that favours the more thermophilic P. cryptogea. The high susceptibility to phytophthora root rot should not be disregarded in olive breeding programmes where wild olive is used as a source of resistance to verticillium wilt.     

Bait twig method for soil detection of <Emphasis Type="Italic">Rosellinia necatrix</Emphasis>, causal agent of white root rot of Japanese pear and apple,at an early stage of tree infection

Mulberry twigs were inserted into the soil as bait to detect Rosellinia necatrix at an early stage of tree infection in the orchard. R. necatrix was frequently trapped on twigs near the trunk base at soil depths of 6–20 cm within 10–20 days in May–July, suggesting that the incubation period was dependent on soil temperature. Subsequently, we inserted twig in the soil around healthy-looking trees in naturally infested orchards. R. necatrix was trapped from 80.0% of Japanese pear and 75.0% of apple trees that later proved to be infected. This bait twig method facilitated quicker diagnosis of white root rot on Japanese pear and apple at early stages of infection and can be used to detect recurrence of the fungus after fungicide treatment.