Phytophthora foot rot control in citrus with Myrothecium roridum1

Foot rot is the most important disease in citrus in Spain. Phytophthora nicotianae var. parasitica is the major cause of this disease. Field treatments with classic contact fungicides have been extensively used for many years to combat stem and trunk Phytophthora diseases in citrus, while, more recently, foliar application of the systemic fungicide fosetyl-A1 has been used with great success. It is now also possible to use a biological control method for foot rot disease. Myrothecium roridum, a cosmopolitan soil hyphomycete with high cellulolytic activity, produces toxins which are very active against Phytophthora fungi. Culture filtrates of M. roridum cultured in liquid medium with low sugar content strongly inhibit in vitro development of P.n. parasitica, P. syringae and P. capsici. In glasshouse trials employingcross inoculations of M. roridum and P.n. parasitica in stems of sweet orange and willowleaf mandarin seedlings, the bark lesions produced by Phytophthora were clearly stopped at 7 or 10 days after inoculation and the cankers subsequently did not show evident activity. In field trials, M. roridum inoculated into the roots of several citrus species limited development of cankers due to P.n. parasitica (artificial or natural infection). The good results open a new way for control of Phytophthora diseases in citrus.     

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.     

First record of Aleurocanthus spiniferus (Homoptera: Aleyrodidae) in Apulia,Southern Italy

The presence of Aleurocanthus spiniferus is reported for the first time in Italy. The orange spiny whitefly was identified in April 2008 in the Apulia region. This is also the first record of this citrus pest in Europe.     

Distribution of Phytophthora spp. in Field Soils Determined by Immunoassay

ABSTRACT Populations of Phytophthora spp. were determined by enzyme-linked immunosorbent assay (ELISA) in field soils used for pepper and soybean production in Ohio. Soybean fields were sampled extensively (64 fields, n = 6 samples per field over 2 years) and intensively (4 fields, n = 64 samples per field in 1 year) to assess heterogeneity of P. sojae populations. Four pepper fields (n = 64), three of which had a history of Phytophthora blight (caused by P. capsici), also were sampled intensively during a 6-month period. Mean (m), variance (v), and measures of aggregation (e.g., variance-to-mean ratio [v/m]) of immunoassay values, translated to Phytophthora antigen units (PAU), were related to the disease history in each of the pepper and soybean fields. Mean PAU values for fields in which Phytophthora root rot (soybean) or blight (pepper) had been moderate to severe were higher than in fields in which disease incidence had been low or not observed. A detection threshold value of 11.3 PAU was calculated with values for 64 samples from one pepper field, all of which tested negative for Phytophthora by bioassay and ELISA. Seven of the eight intensively sampled fields contained at least some detectable Phytophthora propagules, with the percentage of positive samples ranging from 1.6 to 73.4. Mean PAU values ranged from 1 to 84 (extensive soybean field sampling), 6 to 24 (intensive soybean field sampling), and 4 to 30 (intensive pepper field sampling); however, variances ranged from 0 to 7,774 (extensive sampling), 30 to 848 (intensive soybean field sampling), and 5 to 2,401 (intensive pepper field sampling). Heterogeneity of PAU was high in most individual soybean and pepper fields, with values of v/m greater than 1, and log(v) increasing with log(m), with a slope of about 2.0. Spatial autocorrelation coefficients were not significant, indicating there was no relationship of PAU values in neighboring sampling units (i.e., field locations) of the intensively sampled fields. Combined results for autocorrelations and v/m values indicate that Phytophthora was highly aggregated in these fields but that the scale of the aggregation (e.g., average focus size) was less than the size of the sampling units. Because of the observed variability, we calculated that sample sizes of 20 or more would be needed to estimate precisely the mean density of Phytophthora in most cases.     

Protection of Citrus Rootstocks Against Phytophthora spp. with a Hypovirulent Isolate of Phytophthora nicotianae

ABSTRACT Phytophthora root rot of citrus in Florida is caused by Phytophthora nicotianae and P. palmivora. A naturally occurring isolate of P. nicotianae (Pn117) was characterized as hypovirulent on citrus roots. Pn117 infected and colonized fibrous roots, but caused significantly less disease than the virulent isolates P. nicotianae Pn198 and P. palmivora Pp99. Coincident inoculation of rootstock seedlings of Cleopatra mandarin (Citrus reticulata) or Swingle citrumelo (C. paradisi x Poncirus trifoliata) with the hypovirulent Pn117 and the virulent isolates Pn198 and Pp99 did not reduce the severity of disease caused by the virulent Phytophthora spp. When either rootstock was inoculated with the hypovirulent Pn117 for 3 days prior to inoculation with virulent isolates, preinoculated seedlings had significantly less disease and greater root weight compared with seedlings inoculated with the virulent isolates alone. Recovery of the different colony types of Phytophthora spp. from roots of sweet orange (C. sinensis) or Swingle citrumelo was evaluated on semiselective medium after sequential inoculations with the hypovirulent Pn117 and virulent Pp99. Pn117 was isolated from roots at the same level as the Pp99 at 3 days post inoculation. Preinoculation of Pn117 for 3 days followed by inoculation with Pp99 resulted in greater recovery of the hypovirulent isolate and lower recovery of the virulent compared with coincident inoculation.     

Immunodetection of elicitins from Phytophthora spp. using monoclonal antibodies

Ten monoclonal antibodies were selected from mice immunized with a highly purified elicitin secreted by Phytophthora cryptogea , termed cryptogein. These antibodies could be classified into five groups according to their cross-reactivity to heterologous elicitins from other Phytophthora species, from strict specificity (reacting solely with cryptogein) to broad reactivity (reacting with all four elicitins under study). When examined on BIA core (a real-time biospecific interaction analyser), these monoclonal antibodies were found to recognize at least three different epitopes on the cryptogein molecule. Their use in elicitin detection and quantification was optimized in several ELISA protocols. A mixed monoclonal-polyclonal antibody, indirect DAS-ELISA procedure detected as little as 20 pg of purified elicitin per well (100 μl). The four elicitins could be detected with the aid of one of couple of polyvalent reagents, whilst each one could be detected separately using appropriate monoclonal antibodies. These protocols have been used to detect elicitins secreted by Phytophthora spp. into culture medium as well as in planta following plant inoculation.     

Effect of irrigation on the dynamics of Phytophthora citrophthora populations in soil of citrus orchards1

Effects of irrigation on the dynamics of soil populations of Phytophthora citrophthora have been studied in three citrus orchards in East Sicily (Italy). Significant increases of inoculum levels were detected 24 h after irrigation. In one orchard throughout summer, population of P. citrophthora beneath the tree canopy was more than 15 propagules g-¹ soil, a value which is considered a threshold level for root rot infections.     

Synergism between fungicides with different mechanisms of action against acylalanine-resistant strains of Phytophthora spp.1

The authors have studied the efficacy of two systemic compounds, cymoxanil and propamocarb, used as partners of acylalanines, in limiting mycelial growth of acylalanine-sensitive and resistant strains of Phytophthora capsici and P. nicotianae var. parasitica. The fungicidal activity of the single components was compared to that of the different mixtures, at various ratios. The degree of synergy was calculated using the method of Wadley. The results obtained indicate synergistic interactions of various intensities: synergistic factors (SF) up to seven were found. The highest synergistic effects were observed in the case of resistant Phytophthora strains.     

Involvement of Phytophthora cryptogea in sweet cherry decline in Turkey

Sweet cherry is a major commercial crop in Turkey, the most important producer of the fruit worldwide. Sweet cherry decline was observed in an orchard in Ankara province of Turkey. Affected young trees exhibited reduced tree vigor, yellowing and wilting of leaves, and dieback symptoms resulted in tree death. A Phytophthora sp. was consistently isolated from necroses that appeared on taproots and crowns. The pathogen was identified as Phytophthora cryptogea based on several morphological features and DNA sequence of the internal transcribed spacer (ITS) region. P. cryptogea was pathogenic on excised shoots and 1-year-old cherry rootstocks. This is the first report of P. cryptogea causing disease of sweet cherry in Turkey.     

Phytophthora oleae sp. nov. causing fruit rot of olive in southern Italy

A homothallic Phytophthora species was found to be consistently associated with a rot of mature fruits of two local cultivars of olive (Olea europaea) in Calabria, southern Italy. The phylogenetic analysis of sequences of the ITS1‐5.8S‐ITS2 region and cox1 gene enabled its identification as a new species of clade 2, with a basal position compared to previously described subclades. The new species is described formally with the epithet Phytophthora oleae, referring to the natural matrix from which it was isolated. A unique combination of molecular and morphological characters clearly separates P. oleae from other already described Phytophthora species. This new species produced semipapillate, occasionally bipapillate, persistent sporangia on simple sympodially branching sporangiophores as well as globose and smooth‐walled oogonia, paragynous antheridia and spherical, plerotic oospores. The pathogenicity of P. oleae was confirmed in inoculation trials on fruits of three olive cultivars, including the two local cultivars from which the pathogen had been isolated.     

Characterization of Phytophthora spp. Causing Outbreaks of Citrus Brown Rot in Florida

ABSTRACT Epidemics of citrus brown rot from 1994 to 1997 in the south-central and east-coast citrus areas of Florida were characterized and the causal Phytophthora spp. identified. Two species of Phytophthora, P. palmivora and P. nicotianae, were consistently associated with brown rot. Epidemics caused by P. palmivora appeared to be initiated on immature fruit dropped on the orchard floor. The soilborne fungus infected and sporulated on these fruit and was then disseminated to fruit above 1 m in the canopy. In contrast, infection by P. nicotianae, the common cause of root rot, was confined to the lowest 1 m of the canopy. Fruit infected by P. palmivora produced large amounts of ellipsoidal sporangia available for splash dispersal, whereas those infected by P. nicotianae produced far fewer spherical sporangia. Isolates from brown rot epidemics were compared with P. nicotianae from citrus in Florida and Texas, P. citrophthora in California, P. palmivora, and selected Phytophthora spp. from other hosts. Brown rot symptoms produced by the different pathogenic citrus isolates on inoculated fruit were indistinguishable. Morphology, mating behavior, and isozyme patterns of brown rot isolates from 1988 to 1997 matched P. palmivora from citrus roots, other host plants, and other locations, but were different from characterized isolates of P. citrophthora in California and P. nicotianae in Florida and Texas. Cellulose acetate electrophoresis of the isozyme glucose-6-phosphate isomerase rapidly identified the causal citrus pathogen from infected fruit and soil isolation plates. Although P. palmivora is an aggressive pathogen of citrus roots, bark, and fruit, populations in orchard soils were low compared with P. nicotianae.     

诱导疫霉菌产生游动孢子囊液体培养基的研制

研制了几种诱导疫霉菌产生游动孢子囊的蔬菜汁混合培养液,测定了各培养液诱导苎麻疫霉游动孢子囊的效果以及其对苎麻疫霉菌游动孢囊产生量、形态特征、游动孢子释放、卵孢子产生量、卵孢子的形态、菌丝生长和菌落形态等生物学性状的影响。试验结果显示,各配比培养液均能很好地诱导游动孢子囊的产生,且以较低浓度的培养液对孢子囊诱导效果较好,而高浓度则有利于卵孢子的产生,不同培养液中产生的游动孢子囊均能正常释放游动孢子。其中西红柿∶黄瓜=1∶2配比的培养液简单易行且诱导效果最好,是V₈C的理想替代培养液。     

Diclofop-methyl resistance in populations of Lolium spp. from central Italy

Populations of Lolium spp. collected in central Italy were screened for resistance to acetyl-coenzyme A carboxylase (ACCase)-inhibiting herbicides and compared with known susceptible and resistant Lolium rigidum (Gaud.) populations from Australia. Populations Roma'94 and Tuscania'97 were up to 8- and 7.5-fold more resistant to diclofop-methyl, respectively, than susceptible populations in pot experiments. However, populations Tarquinia'97 and Vetralla'94 were not resistant. Diclofop-methyl resistance levels in the Italian populations were lower than in the Australian populations SLR31 and WLR96 (16.5 and > 64 times more resistant than S respectively). In an agar germination test, Tuscania'97 showed low levels of cross-resistance to fluazifop-p-butyl, whereas no cross-resistance was found in the Roma'94 population.     

Clostridium bifermentans and C. subterminale are associated with kiwifruit vine decline,known as moria,in Italy

Since 2012, a new pathogenic syndrome has frequently been observed in many areas of kiwifruit cultivation in Italy. The main symptoms include an initial withering of the leaves followed by a total and sudden collapse of plants, mainly occurring during summer. The withered leaves fall and the main and secondary feeder roots appear rotten, sometimes showing a reddish-brown discoloration. The disease, that affects both the green and yellow-fleshed cultivars, has been called kiwifruit vine decline and is locally known as moria. The syndrome has been found consistently associated with soil waterlogging, which frequently occurs either after the traditional agronomical practice of irrigating orchards through surface irrigation or after very heavy rainfall. So far, the role played by bacteria in this syndrome has not been investigated. In the present study, Clostridium spp. were isolated from both rotten roots and soils obtained from Italian kiwifruit orchards affected by the syndrome, indicating for the first time that anaerobic bacteria are able to cause damage to woody crops. C. bifermentans and C. subterminale incited symptoms in kiwifruit in both in vivo and in vitro pathogenicity tests. Soil waterlogging seems to potentially favour colonization of kiwifruit roots by anaerobic bacteria, probably because saturation of the soil can facilitate proliferation and persistence of these bacteria during long periods of the vegetative growth of the crop. The occurrence of anaerobic bacteria does not exclude the possibility that other microorganisms can play additional/synergic role(s) in causing the kiwifruit vine decline.     

中国柑橘黑腐病和褐斑病病原菌的系统发育分析

 近些年链格孢属真菌的分类有了长足的进展,但柑橘褐斑病和黑腐病的病原却还是存在着一些争议。本研究从中国柑橘果实上收集了49株黑腐病菌菌株,并选取了8株从柑橘叶片上分离的具代表性的褐斑病菌菌株,基于Alta1 、endoPG、LSU、OPA10-2、OPAl-3和OPA2-1 等6个基因位点串联序列构建系统发育树。结果表明,柑橘黑腐病和褐斑病都可由不止一种链格孢菌引起,均以Alternaria alternata为主。两种病害的病原菌之间不能通过该系统发育树区分,但在致病性上存在差异,且能通过扩增ACT毒素合成基因进行区分。为了使两种病害的病原更加方便阐述,作者建议以它们的主要类群对其进行命名,柑橘褐斑病病原学名还是遵循前人的称呼,为the tangerine pathotype of A. alternata,即链格孢菌橘致病型,而柑橘黑腐病病原应为A. alternata,即链格孢菌。     

中国柑橘黑腐病和褐斑病病原菌的系统发育分析

近些年链格孢属真菌的分类有了长足的进展,但柑橘褐斑病和黑腐病的病原却还是存在着一些争议。本研究从中国柑橘果实上收集了49株黑腐病菌菌株,并选取了8株从柑橘叶片上分离的具代表性的褐斑病菌菌株,基于Alta1、endoPG、LSU、OPA10-2、OPAl-3和OPA2-1等6个基因位点串联序列构建系统发育树。结果表明,柑橘黑腐病和褐斑病都可由不止一种链格孢菌引起,均以Alternaria alternata为主。两种病害的病原菌之间不能通过该系统发育树区分,但在致病性上存在差异,且能通过扩增ACT毒素合成基因进行区分。为了使两种病害的病原更加方便阐述,作者建议以它们的主要类群对其进行命名,柑橘褐斑病病原学名还是遵循前人的称呼,为the tangerine pathotype of A.alternata,即链格孢菌橘致病型,而柑橘黑腐病病原应为A.alternata,即链格孢菌。     

Observations of Phytophthora spp. in Water Recirculation Systems in Commercial Hardy Ornamental Nursery Stock

Samples of water and sediment were taken from drains, reservoirs and wells from four commercial hardy ornamental nurseries with water recirculation systems. The samples were taken on seven different dates throughout a single year from August 1994 to July 1995. The samples were screened for Phytophthora species using five different methods: direct plating, three bait tests (using lupin seedlings, apples and Rhododendron leaves) and a DAS-ELISA (double-antibody sandwich enzyme-linked immunosorbent-assay) with two antisera. In the nurseries with old water recirculation systems, Phytophthora species were detected in the drains and in the reservoirs. In the nursery with a new recirculation system, the pathogens were only present in the drains. None of the water samples from wells in any of the nurseries were contaminated. Phytophthora species were present in the water as well as in the sediment samples from drains and reservoirs. They were detected in the water recirculation systems irrespective of the season. The number of isolates increased about sevenfold between late summer and spring. At least 12 different Phytophthora species were identified: some isolates were previously unrecorded species. The epidemiology of the pathogens in outdoor water recirculation systems as well as the importance of the results for commercial nurseries is discussed.