Differences in Strain Virulence of the European Stone Fruit Yellows Phytoplasma and Susceptibility of Stone Fruit Trees on Various Rootstocks to this Pathogen

Twenty strains of the European stone fruit yellows (ESFY) phytoplasma showed great differences in virulence when examined by graft inoculation of trees on peach, peach hybrid GF 677 and P. 'Marianna' GF 8/1 rootstocks. The most virulent strains killed all trees on peach rootstocks whereas the mild strains did not cause mortality but induced only mild foliar symptoms and slightly reduced vigour. Virulence often depended on the pathogen–scion combination and was in several cases most severe when the scion consisted of the original host of the pathogen. To examine resistance in stone fruits, trees on a total of 23 rootstocks were inoculated with the ESFY strains. Trees on the Prunus domestica stocks Ackermann's, Brompton and P 1275 and on Prunus cerasifera stock Myrabi were little affected. Slightly more damage occurred in trees on rootstocks GF 677, GF 8–1, and the P. insititia stocks St Julien A and St Julien GF 655/2. Ishtara, P. cerasifera stock Myrobalan, and peach rootstocks Higama and GF 305 were shown to be moderately susceptible and a high susceptibility was found in trees on peach rootstocks Montclar, peach seedling, Rutgers Red Leaf, and Rubira, on apricot seedlings and St Julien 2. Of flowering cherry trees on various rootstocks, the least susceptible were those on Gisela 3 and F 12/1 whereas Gisela 1, Weihroot 158 and Gisela 5 were more affected. Phytoplasmas were detected by either DAPI (4'-6-diamidino-2-phenylindole) staining or polymerase chain reaction in all rootstocks and scions tested. However, detection frequency and phytoplasma concentrations were usually lower in the more tolerant hosts than in susceptible genotypes.     

Relative Resistance of Four Peach Rootstocks to Phytophthora cactorum and Phytophthora megasperma

Shoot samples of four peach rootstocks that are important to the peach industry in Greece: KID I, GF305, GF677, and PR204, were inoculated with Phytophthora cactorum and Phytophthora megasperma in the field and in the glasshouse and were then evaluated with regard to susceptibility. The pathogenicity of P. cactorum and P. megasperma to peach rootstocks was confirmed with an excised twig assay. The peach rootstocks showed differential susceptibility to P. cactorum . GF305 was the least susceptible and KID I was the most susceptible, which suggests that the latter rootstock is unsuitable for orchards in which the conditions are favourable for Phytophthora diseases. GF677 and PR204 were moderately susceptible. The plants that were inoculated with P. megasperma in the field and in the glasshouse showed no sign of infection whereas twigs inoculated in vitro with P. megasperma developed necrosis. GF305 was the most resistant, KID I was the most susceptible and PR204 and GF677 were moderately resistant. The present results demonstrate that none of the four peach rootstocks used in this study was completely resistant to P. cactorum , particularly when rootstocks were flooded periodically to enhance disease development. Therefore an integrated approach including host and resistance cultural practices is recommended to manage diseases caused by P. cactorum in peach orchards in Greece.     

大豆疫霉根腐病抗源筛选

由大豆疫霉菌引起的大豆疫霉根腐病是大豆生产的重要病害,该病已在我国大豆主要产区发生,并在局部地区造成较大产量损失。利用抗病品种是防治大豆疫霉根腐病最有效的方法。本研究目的是筛选大豆疫霉根腐病抗源,为病害防治和抗病品种的选育提供参考。用下胚轴创伤接种方法对120个栽培大豆品种（系）进行接种,鉴定其对10个具有不同毒力大豆疫霉菌菌株的抗性。有110个品种（系）分别抗1～10个大豆疫霉菌菌株,其中以河南大豆品种（系）对疫霉菌的抗性最丰富,安徽、湖北和山西大豆品种（系）也具有抗性多样性。120个大豆品种（系）对10个大豆疫霉菌菌株共产生57个反应型,有4个抗性反应型分别与单个抗病基因的反应型一致,有7个抗性反应型与2个已知基因组合的反应型相同,其他抗性反应型为新的类型。一些大豆品种（系）中可能存在有效的抗大豆疫霉根腐病新基因。     

Phytophthora Crown and Root Rot of Apricot Caused by Phytophthora palmivora in Turkey

Forty‐nine Phytophthora isolates were obtained from roots and crown of apricot trees with symptoms of decline grown in commercial orchards in Malatya, Elaz?? and Diyarbak?r provinces, Turkey, in 2011 and 2013. All of the recovered isolates were identified as Phytophthora palmivora on the basis of morphological characteristics. Blast analysis of ITS region sequences of rDNA of 5 isolates revealed 100% identity with a reference isolates of P. palmivora from GenBank. Isolates of P. palmivora were pathogenic on 12‐month‐old wild apricot rootstock ‘Zerdali’ plants that were wound inoculated on the roots and on the crown. This study demonstrated that P. palmivora is the cause of the crown and root rot found on apricot in Turkey. To our knowledge, this is the first report of P. palmivora on this host plant.     

抗大豆疫霉根腐病野生大豆资源的初步筛选

由大豆疫霉菌引起的大豆疫霉根腐病是严重影响大豆生产的毁灭性病害之一.防治该病唯一经济、有效和环境安全的方法是利用抗病品种.本研究对野生大豆资源进行抗大豆疫霉根腐病初步筛选,以期探讨野生大豆的抗性水平、分布和获得抗性野生大豆资源.通过苗期接种大豆疫霉菌对412份野生大豆资源进行抗病性鉴定,有13.4%的资源抗大豆疫霉根腐病,15.3%的资源表现为中间反应类型.对野生大豆资源的来源分析表明,抗大豆疫霉根腐病野生大豆资源在我国分布广泛,其中安徽省野生大豆资源抗性最丰富.     

基于SSR标记分析大豆疫霉根腐病抗源的遗传多样性

丰富的遗传多样性可为大豆育种提供宽阔的遗传基础,本研究基于35对SSR标记,对60份东北地区大豆疫霉根腐病抗性品种进行了遗传多样性分析,共检测到189个等位基因,平均每个位点等位变异数5.4个,多态性信息含量指数(PIC)为0.1550~0.8195,平均为0.6636;遗传相似系数的变异范围为0.31~0.74。利用5对高多态性SSR引物构建了60份抗性材料的指纹图谱,这5对SSR引物构建的指纹图谱可以将60份疫霉根腐病抗性材料逐一区分开。采用NTSYS2.10基于遗传距离的聚类分析,将60份抗性材料分为7个类群,其中78.33%的抗性品种(系)的遗传相似系数在0.45~0.74间,表明遗传差异相对较窄,品种间遗传多样性水平较低。聚类分析与群体遗传结构分析结果有部分重合,均反映出不同地区的抗性材料间存在一定的渗透和交流。     

Phytophthora cinnamomi Associated with Root and Crown Rot of Walnut in Turkey

Walnut decline caused by Phytophthora sp. occurred in an orchard in Sakarya province in Turkey. Affected young trees showed poor growth, leaf discolouration, root and crown rot and eventual death. A Phytophthora sp. isolated from necrotic taproots and crown tissues. The causal agent of the disease was identified as Phytophthora cinnamomi by morphological characteristics and comparing sequences of internal transcribed spacer (ITS) region. Upon conducting pathogenicity test, averaging 7.8‐cm‐long canker developed on basal stem within 2 weeks, while no cankers developed in the control plants.     

梨品种资源果实轮纹病抗性的评价

采用田间人工接种的方法,2009-2010年连续两年对保存于国家梨种质资源更新圃的182个品种进行果实轮纹病抗性评价。结果表明：砂梨、白梨、秋子梨、种间杂交梨、西洋梨和新疆梨的两年平均发病率分别为6.15%、7.20%、7.43%、12.66%、17.00%、18.93%,梨品种间对轮纹病抗性存在差异,不同条件下多数品种轮纹病发病率有差异。依据两年平均发病率进行抗性评价分级,72个品种为高抗、63个品种为抗、25个品种为中抗、14个品种为低抗、8个品种为不抗。     

小麦种质对茎基腐病抗性评价及优异种质筛选

小麦茎基腐病是由镰孢菌侵染引起的一种世界性土传病害,近年来已严重威胁到我国小麦的安全生产。为筛选具有茎基腐病抗性的小麦种质资源,本研究采用孢子悬浮液浸种法,分别以国外抗病材料Sunco和中国品种陕253为抗病和感病对照,对670份我国小麦品种(系)进行了茎基腐病温室苗期抗性鉴定。结果发现,我国供试品种(系)感病材料(病情指数>30)所占比例达到84%,且包含多个近年来小麦生产中的主推品种,表明我国小麦品种总体抗性水平低是导致茎基腐病近年来发病频率与程度不断增加的重要原因之一。经多轮筛选,发掘获得15份抗病表现稳定、抗性水平与抗病对照Sunco相仿的材料。15份材料平均病情指数在10.9~19.4之间,其株高、抽穗期等农艺性状表现出较为丰富的变异,为我国小麦抗茎基腐病品种选育和抗性遗传研究提供了种质资源。     

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.     

豌豆茎基腐病病原菌分离鉴定及其对杀菌剂的敏感性测定

豌豆(Pisum sativum)是我国重要的豆类经济作物, 病害对豌豆生产造成重大经济损失。通过形态学观察、分子鉴定以及致病性测定, 最终确定引起豌豆茎基腐病的3种病原菌分别为尖孢镰刀菌(Fusarium oxysporum)、芸苔链格孢菌(Alternaria brassicae)和格氏镰刀菌(F. grosmichelii), 优势菌株为尖孢镰刀菌, 分离率为53.6%。室内毒力测定结果表明, 5种供试杀菌剂对3种病原菌的菌丝生长均有抑制作用, 其中咯菌腈和戊唑醇的抑菌效果最好。研究结果为豌豆茎基腐病的防治提供了科学依据。     

Nested PCR Assays for Detection of Monilinia fructicola in Stone Fruit Orchards and Botryosphaeria dothidea from Pistachios in California

Nested polymerase chain reaction (PCR) assays were developed based on microsatellite regions for detection of Monilinia fructicola, the causal agent of brown rot of stone fruits, and Botryosphaeria dothidea, the causal agent of panicle and shoot blight of pistachio. The nested PCR primers specific to M. fructicola were developed based upon the sequence of a species‐specific DNA fragment amplified by microsatellite primer M13. The external and internal primer pairs EMfF + EMfR and IMfF + IMfR amplified a 571‐ and a 468‐bp fragment, respectively, from M. fructicola, but not from any other fungal species present in stone fruit orchards. The nested PCR primer pairs specific to B. dothidea were developed based upon the sequence of a species‐specific 1330‐bp DNA fragment amplified by microsatellite primer T3B. The external and internal primer pairs EBdF + EBdR and IBdF + IBdR amplified a 701‐ and a 627‐bp fragment, respectively, from B. dothidea, but not from any other fungal species associated with pistachio. The nested PCR assays were sensitive enough to detect the specific fragments in 1 fg of M. fructicola or B. dothidea DNA or in the DNA from only two conidia of M. fructicola or B. dothidea. The nested PCR assays could detect small numbers of M. fructicola conidia caught on spore‐trap tapes and detect visible infections of B. dothidea in pistachio tissues. Microsatellite regions with high numbers of copies are widely dispersed in eukaryotic genomes. The results of this study indicate that microsatellite regions could be useful in developing highly sensitive PCR detection systems for phytopathogenic fungi.     

Effects of Metalaxyl, Fosetyl-Al, Dimethomorph and Cymoxanil on Phytophthora cactorum of Peach Tree

The activity of metalaxyl, fosetyl-Al, dimethomorph, and cymoxanil against Phytophthora crown rot of peach trees was examined. Application of fosetyl-Al or metalaxyl by painting the trunk (150 g/l) of 3-year-old PR204 trees was inhibitory to growth of the fungus but neither dimethomorph nor cymoxanil were effective. Application of metalaxyl as a soil drench suppressed canker development when the treated trees were subsequently inoculated with Phytophthora cactorum . Fosetyl-Al significantly reduced the growth of fungus compared with cymoxanil, dimethomorph and control but it was not as effective as metalaxyl. Again, dimethomorph and cymoxanil did not influence canker development. Both metalaxyl and fosetyl-Al were active for at least 21 days after applications. Strips of trunk bark were removed from trees, that had been drenched with the tested chemicals 20 days before, and inoculated on the cambium side with P. cactorum . Metalaxyl was the most effective fungicide and fosetyl-Al significantly reduced the development of fungus compared with dimethomorph, cymoxanil and the untreated strips. Colonization of strips treated with dimethomorph was significantly less than untreated strips. In contrast, cymoxanil did not inhibit the growth of fungus. Application of metalaxyl or fosetyl-Al as trunk paint or a soil drench appear to be effective procedures for preventing Phytophthora rot of peach trees.     

大豆品种早熟18抗疫霉根腐病基因的SSR分子标记

大豆品种早熟18是抗疫霉根腐病的有效抗源。本研究鉴定和分子标记早熟18的抗疫霉根腐病基因,以期为该品种的有效利用及分子辅助育种奠定基础。以感病大豆品种Williams与早熟18杂交建立分离群体。抗性遗传分析表明,早熟18对大豆疫霉菌抗性由1个显性单基因控制,该基因被定名为RpsZS18。SSR标记连锁分析表明,RpsZS18位于大豆分子遗传连锁群D1b上的SSR标记Sat_069和Sat_183之间,与这两个标记的遗传距离分别为10．0cM和8．3cM。RpsZS18是D1b连锁群上鉴定的第一个抗疫霉根腐病基因。     

大豆种质对疫霉根腐病抗性特点研究

对1027份中国和国外引进的大豆种质进行了大豆疫霉(Phytophthora sojae)根腐病的抗病性鉴定评价.结果表明,中国大豆种质的抗病性高于国外引进种质;中国南方的大豆种质抗病性较北方种质强,长江流域大豆中抗病种质比率最高,其次为黄淮海流域种质,而东北地区抗病种质较少;不同省份大豆种质的总体抗病性差异明显;育成品系的抗性好于改良品种和农家种,但不同省份来源的农家种、品系和品种抗性存在差异,黑龙江材料抗病性最低,这也是该省大豆疫霉根腐病严重发生的重要原因之一;在大豆籽粒脐色为黄色和褐色的材料中,抗病种质较多.     

野生大豆资源对大豆疫病抗病性和耐病性鉴定

大豆疫病是大豆重要病害之一,在世界范围内导致严重经济损失。防治大豆疫病最有效方法是利用抗病或耐病品种。筛选抗性资源是发掘抗性基因和抗病育种的基础。本研究鉴定了野生大豆资源对大豆疫病的抗病性和耐病性,以期发掘优异抗源。苗期用子叶贴菌块方法鉴定104份野生大豆资源对两个不同毒力的大豆疫霉分离物PSJS2(毒力型:1a,1b,1c,1d,1k,2,3a,3b,3c,4,5,6,7,8)和PS41-1(毒力型:1a,1d,2,3b,3c,4,5,6,7,8)抗性,结果表明33份资源抗PS41-1,35份资源抗PSJS2,其中18份抗两个分离物。在抗病性鉴定基础性上,用菌层接种方法对选择的82份资源进行耐病性鉴定,发现7份高耐病性资源。这些结果表明,野生大豆中可能含有新的大豆疫病抗病和(或)耐病资源,这些抗病或耐病资源可以用于未来大豆抗病育种,以丰富大豆对大豆疫病的抗性遗传基础。     

Evaluation of fungicides and biocontrol products for the control of Phytophthora root rot of hydrangeas

Abstract

Phytophthora root rot is one of the most important diseases in almost all hydrangeas of nursery production. In this study, the efficacy of fungicides and biocontrol products against Phytophthora root rot of hydrangea was assessed in greenhouse and field experiments. Treatments used in field or greenhouse experiments were RootShield PLUS⁺, MBI110, IT-5103, Grotab, OxiPhos, TerraClean 5.0?+?TerraGrow program, Segovis, Pageant Intrinsic, Empress Intrinsic and Subdue Maxx. Pots/plots were inoculated with Phytophthora nicotianae grown on rice grains, sterilised rice grains were used for negative controls. After the trials, plant growth data (total plant weight, root weight, plant height, plant width) were recorded, and roots were assessed for disease severity using a scale of 0–100%. The treatments most effective in reducing Phytophthora root rot severity were Segovis, Empress Intrinsic, Subdue Maxx, TerraClean 5.0?+?TerraGrow program in both greenhouse and field experiments. This study will help nursery producers make proper management decisions by using recommended fungicides and biocontrol products of this study in a rotation or alone to manage Phytophthora root rot of hydrangea.     

The relationship between colonisation and crown rot symptoms in strawberry plants infected with Phytophthora cactorum

Strawberry tissues infected with Phytophthora cactorum were comminuted and plated in a selective antibiotic agar medium to determine levels of tissue colonisation as indicated by the number of colony forming units (CFU) recovered per gramme of infected tissue. The number of CFU recovered per gramme of tissue increased logarithmically with the amount of necrosis in infected crown, leaf and petiole tissues. Under the conditions of enhanced susceptibility to infection and colonisation caused by cold storage treatments, this relationship between colonisation and necrosis was not significantly altered in the susceptible cv. Tamella. A recovery index was used to determine the effect of infected tissues on the recovery of CFU. This indicated that increasing levels of host colonisation stimulated CFU recovery and may partly explain the large increase in CFU g^-1 with larger amounts of necrosis. The amount of tissue colonisation was greater in inoculated plants of the susceptible cv. Tamella than in less susceptible cv. Cambridge Favourite, although the necrotic tissues of the latter contained more CFU g^-1, indicating a greater level of tolerance to colonisation. In cv. Tamella small amounts of colonisation were capable of causing wilt symptoms, although no wilted plants contained less than 200 CFU g^-1. Conversely, plants containing more than 1000 CFU g^-1 always wilted. In the early stages of infection, low levels of colonisation could be detected in strawberry crowns in the absence of symptoms. Dormant strawberry plants of cv. Tamella were readily infected by P. cactorum zoospore inoculations but, unlike actively growing plants, the majority of infections remained latent. These latent infections exhibited little or no symptoms and CFU recoveries from infected tissues were always below 100 CFU g^-1.     

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.     

Biological Control of Phytophthora Root Rot of Pepper Using Trichoderma harzianum and Streptomyces rochei in Combination

A combination of two compatible micro‐organisms, Trichoderma harzianum and Streptomyces rochei, both antagonistic to the pathogen Phytophthora capsici, was used to control root rot in pepper. The population of the pathogen in soil was reduced by 75% as a result. Vegetative growth of the mycelium of P. capsici was inhibited in vitro on the second day after P. capsici and T. harzianum were placed on the opposite sides of the same Petri plate. Trichoderma harzianum was capable of not only arresting the spread of the pathogen from a distance, but also after invading the whole surface of the pathogen colony, sporulating over it. Scanning electron microscopy showed the hyphae of P. capsici surrounded by those of T. harzianum, their subsequent disintegration, and the eventual suppression of the pathogen's growth. Streptomyces rochei produced a zone of inhibition, from which was obtained a compound with antioomycete property secreted by the bacteria. When purified by high‐pressure liquid chromatography, this compound was identified as 1‐propanone, 1‐(4‐chlorophenyl), which seems to be one of the principal compounds involved in the antagonism. A formulation was prepared that maintained the compound's capacity to inhibit growth of the pathogen for up to 2 years when stored at room temperature in the laboratory on a mixture of plantation soil and vermiculite. The two antagonists, added as a compound formulation, were effective at pH from 3.5 to 5.6 at 23–30°C. The optimal dose of the antagonists in the compound formulation was 3.5 × 10⁸ spores/ml of T. harzianum and 1.0 × 10⁹ FCU/ml of S. rochei. This is the first report of a compound biocontrol formulation of these two antagonists with a potential to control root rot caused by P. capsici.