南宁市朱槿曲叶病毒病病原分子鉴定和寄主范围研究

［目的］ 明确南宁市朱槿曲叶病毒病的病原及其寄主范围,初步了解各朱槿品种的抗病性。［方法］ 对采自南宁市区的朱槿病株分离物进行分子鉴定及寄主范围测定;进行朱槿曲叶病毒病发生情况调查,根据发病率高低和症状轻重对南宁市常见的朱槿品种抗病性进行初步评价。［结果］ 基因克隆与序列比较结果表明,该病的病原为木尔坦棉花曲叶病毒（Cotton leaf curl Multan virus, CLCuMV）,其DNA A全长含2 737个核苷酸,伴随有卫星DNAβ分子,分子大小为1 346个核苷酸;研究发现该病毒可以通过B型烟粉虱传播,烟粉虱接种和PCR检测试验〖JP2〗结果表明,该病毒可侵染朱槿、红麻、棉花、西菲葵、黄蜀葵等5种供试的锦葵科植物,表现典型曲叶症状;初步调查评价结果,‘朱砂红朱槿’、‘泰国黄朱槿’、‘重粉朱槿’、‘锦球朱槿’和‘马旦朱槿’属于高度感病品种;‘阿美丽坚朱槿’、‘乳斑朱槿’属于感病品种;‘大红朱槿’、‘大红花’和‘粉喇叭’属于中抗品种;‘红龙朱槿’、‘红喇叭’、‘青杆吊钟’和‘七彩朱槿’属于高抗品种。［结论］ 上述结果可为有效控制朱槿曲叶病毒病提供依据。     

Characterization and host range evaluation of Pseudomonas viridiflava from melon, blite, tomato, chrysanthemum and eggplant

The fluorescent bacterium Pseudomonas viridiflava (Burkholder) Dowson was identified as the causal agent of bacterial leaf blight of melon, tomato, blite, eggplant and of pith necrosis of chrysanthemum plants. Koch's postulates were fulfilled on greenhouse-grown or on potted plants under controlled environmental conditions. Twenty seven cultivated and one weed species, when artificially inoculated, were found to be susceptible to a strain of the bacterium isolated from cucumber. In cross-inoculation tests on potted plants, each strain induced the disease symptoms, independently of the host of origin. To our knowledge, melon and blite have not been previously reported as natural hosts of P. viridiflava. Also this is the first record of the bacterium as a foliar pathogen of melon, tomato, eggplant and blite and as a stem pathogen of chrysanthemum in Greece.     

Tomato leaf curl geminivirus in Australia: occurrence, detection, sequence diversity and host range

The occurrence of whitefly transmitted geminiviruses in Australia was studied using a mixed DNA probe capable of detecting a range of distinct geminiviruses. The only geminivirus species detected was Tomato leaf curl virus (TLCV), which is spread across a vast geographical region of far-northern coastal Australia, an area inhabited by the Australasian-Oceania biotype of Bemisia tabaci . The newly introduced silverleaf whitefly, B. tabaci biotype B, forms high population densities in the eastern coastal region of Queensland and is currently located approximately 150 km from the nearest known TLCV-infected area. The viral host range appeared to be narrow and of 58 species of crop plants and weeds inoculated using the B biotype, only 11 became infected with the virus, including five that did not show foliar symptoms. A DNA fragment of 694 nt, including the complete C4 open reading frame (ORF), the overlapping N-terminal part of the C1 ORF and the viral iterons involved in replication, was amplified from 11 TLCV field isolates and sequenced. Sequence analysis revealed an overall sequence variation of up to 14% in this region, as well as the presence of distinct viral iterons.     

肾形肾状线虫（Rotylenchulus reniformis）浙江杭州 群体形态学和分子特性及寄主范围

 肾形肾状线虫（Rotylenchulus reniformis）是一种重要的蔬菜线虫,寄主范围和分布很广。本文根据形态学特征和形态测量将来自浙江省杭州市蔬菜上的肾状线虫群体鉴定为肾形肾状线虫,为浙江省首次报道。该群体的形态学特征及形态测量值与模式标本基本吻合,个别形态测量值(比值)有变异。通过对该肾形肾状线虫群体的rDNA中ITS区和编码28S RNA基因中的D2D3区的PCR扩增和序列测定,明确二序列长度分别为808 bp和786 bp;序列分析发现该群体ITS序列与肾形肾状线虫种内一些两性生殖群体的亲缘关系较近,同源性高达99.5%～100%,而与一孤雌生殖群体同源性为92.4%;编码28S RNA基因中的D2D3区与已报道群体的同源性达到99.4%,相似度较高;本研究还发现杭州肾形肾状线虫群体的寄主涉及11个科16个属的果蔬。     

Evaluating aggressiveness and host range of Alternaria dauci in a controlled environment

The aggressiveness of Alternaria dauci isolates was investigated in greenhouse conditions. Twenty‐seven isolates were pre‐selected from a large collection to represent high diversity according to geographic or host origins and intergenic spacer (IGS) polymorphism. IGS sequence analysis revealed that isolates were grouped within three different clusters. Eleven isolates were selected and inoculated on a susceptible carrot cultivar. Three criteria (mean lesion number, mean necrotic leaf area and mean disease index) were used to assess the aggressiveness of isolates. Continuous variation in aggressiveness was shown and no clear division into isolate classes was evident. For the host range study, two isolates were inoculated under greenhouse conditions onto nine cultivated Apiaceae species, two wild Daucus species and six cultivated non‐Apiaceae species representing six botanical families. Lesions varying in severity were observed on all dicot species (Apiaceae and non‐Apiaceae), but no symptoms developed on the two monocots studied (leek and sweetcorn). Plant species were also differentiated on the basis of expanding lesions (cultivated and wild carrot, dill and fennel) or non‐expanding lesions (other dicot species). Typical A. dauci conidia were observed after in vitro incubation of leaves with symptoms. Fungal structures were isolated from lesions and A. dauci was confirmed on the basis of conidial morphology and specific conventional PCR results. Genotyping of individual isolates performed with microsatellite markers confirmed the presence of the inoculated isolate. The results clearly showed that, in controlled conditions, the host range of A. dauci is not restricted to carrot.     

加拿大蓟天敌─欧洲方喙象的寄主专一性测定

在温度为25-28℃的养虫室内选择具有代表性的菊科植物及豆科、禾本科牧草共10科30种植物对欧洲方喙象进行寄主专一性测定试验,结果表明其食性专一,只取食加拿大蓟,个别成虫或幼虫在饥饿状态下仅少量取食菊科其他植物,但随后即拒绝取食。非选择性试验(饥饿试验)表明欧洲方喙象耐饥饿能力强,不同龄期平均达28d。欧洲方喙象最适宜的生长温度是25℃,高于30℃或低于10℃,生长发育则受到抑制。     

加拿大蓟天敌─欧洲方喙象的寄主专一性测定

在温度为25～28 ℃的养虫室内选择具有代表性的菊科植物及豆科、禾本科牧草共10科30种植物对欧洲方喙象进行寄主专一性测定试验，结果表明其食性专一，只取食加拿大蓟，个别成虫或幼虫在饥饿状态下仅少量取食菊科其他植物，但随后即拒绝取食。非选择性试验（饥饿试验）表明欧洲方喙象耐饥饿能力强，不同龄期平均达28 d。欧洲方喙象最适宜的生长温度是25 ℃，高于30 ℃或低于10 ℃，生长发育则受到抑制。     

Molecular,morphological and pathogenic diversity of Sclerotinia sclerotiorum isolates from common bean (Phaseolus vulgaris) fields in Argentina

White mould, caused by Sclerotinia sclerotiorum, is one of the most threatening fungal diseases occurring across major bean production regions worldwide. In Argentina, under favourable weather conditions, up to 100% seed yield losses occur on susceptible common bean cultivars. The aim of this study was to characterize the diversity of S. sclerotiorum isolates from six dry bean fields in the main production area of Argentina by means of molecular, morphological (mycelium colour, number and pattern of sclerotia distribution) and pathogenic approaches. Among 116 isolates analysed, high genotypic and morphological variability was observed. A total of 52 mycelial compatibility groups (MCGs) and 59 URPs (universal rice primers) molecular haplotypes were found. All the MCGs were location specific, while only 12% of the URP haplotypes were shared among locations. The molecular analysis of variance revealed a significant differentiation among populations, with higher genetic variability within the populations analysed than among them. The aggressiveness of the isolates towards bean seedlings was assessed in the greenhouse. Most of the isolates were highly aggressive, while no variation among locations was observed. The information generated in the present study provides, for the first time, information on the variability of S. sclerotiorum associated with white mould in the main common bean production area in Argentina. In addition, the findings suggest the occurrence of both clonal and sexual reproduction in the populations analysed. This work contributes to the development of sustainable management strategies in bean production aimed to minimize yield losses due to white mould.     

Morphological,molecular phylogenetic and pathogenic analyses of Erysiphe spp. causing powdery mildew on cashew plants in Brazil

Cashew powdery mildew is presently the most important disease of cashew trees in all Brazilian growing regions. Although it was described over a century ago, it had never threatened the Brazilian cashew industry until the first decade of the 21st century. Morphological and pathogenic evidence indicated the possibility of different pathogen species being involved in early and late types of cashew powdery mildew. This study was designed to elucidate this issue by comparing two different powdery mildew fungi occurring on cashew plants in Brazil according to the morphological characteristics, phylogenetic relationships with closely related powdery mildew fungi and pathogenic relationships. Based on morphology, molecular phylogenetics and pathogenicity on cashew, it was shown that two species of powdery mildew specimens are without question associated with cashew trees. One species, which infects young immature tissues such as shiny leaves, flowers and young fruits, is Erysiphe quercicola, while Erysiphe necator is associated exclusively with mature leaves. This is the first report of both E. quercicola and E. necator causing cashew powdery mildew, and the first detection of E. necator on cashew.     

裸仁美洲南瓜叶枯病病原菌鉴定及其寄主范围和侵染条件

近年来,在甘肃省武威地区裸仁美洲南瓜发生了一种叶枯病,采用常规组织分离法对罹病组织进行了病原菌分离培养,以及病原菌的形态、分离培养性状观察和致病性测定.结果表明,引起裸仁美洲南瓜叶枯病的致病菌为瓜链格孢Alternaria cucumerina(E11.et Ev.)E11iott.寄主范围测定表明,该病原菌可侵染所有供试葫芦科植物,其中南瓜、西葫芦和甜瓜发病最严重;丝瓜和苦瓜发病较轻;该病原菌还可引起白菜和菜豆发病.温度和保湿时间对病原菌的侵染影响较大,25～30℃温度范围内最易侵染,病害潜育期仅为2～3天;当保湿时间60h时,病叶率达到88%,而保湿时间为6h时,病叶率仅为8%.南瓜不同品种对叶枯病的抗性差异较大,其中二星、天然、光板1号、无壳(李建民)和金无壳为高抗品种;四星、粒赛金、金大地、郑峰光板、日本南瓜为高感品种.     

Distribution,host range and some properties of a virus disease of sunflower in Kenya

Abstract

Apparently similar viruses were associated with two different disease symptoms in sunflower (Helianthus annuus) in Kenya, one a yellow blotch and the other a leaf crinkle. The virus occurred in all sunflower‐growing regions but the crinkle disease was confined to areas east of the rift valley. Disease incidence varied with locality and season. Mechanical transmission of virus from plants with either disease was possible, but the yellow blotch virus was also transmissible from an important alternate field host, Tridax procumbens by Aphis gossypii to 23 species in three families. Partially purified preparations of virus made from mechanically inoculated seedlings affected with either symptom type contained 26 nm virus particles which, in ISEM tests, reacted with antisera prepared against six members of the luteovirus group.     

Characterization of Passionfruit severe leaf distortion virus,a novel begomovirus infecting passionfruit in Brazil,reveals a close relationship with tomato‐infecting begomoviruses

Molecular and biological characterization of the begomovirus isolate BR:LNS2:Pas:01, obtained from yellow passionfruit plants in Livramento de Nossa Senhora, Bahia state, Brazil, was carried out. Sequence analysis demonstrated that the BR:LNS2:Pas:01 DNA‐A had highest nucleotide sequence identity with Tomato chlorotic mottle virus (77%) and had five ORFs corresponding to the genes cp, rep, trap, ren and ac4. The DNA‐B had highest nucleotide sequence identity with Tomato yellow spot virus (74%) and two ORFs corresponding to the genes mp and nsp. These identity values indicate that this isolate represents a new begomovirus species, for which the name Passionfruit severe leaf distortion virus (PSLDV), is proposed. Phylogenetic analysis clustered the PSLDV DNA‐A and ‐B in a monophyletic branch with Brazilian tomato‐infecting begomoviruses. The isolate’s host range was restricted to species from the Passifloraceae and Solanaceae. PSLDV‐[BR:LNS2:Pas:01] was capable of forming pseudorecombinants with tomato‐infecting begomoviruses, reinforcing its close relationship with these viruses and suggesting a possible common origin. However, the virus was not capable of infecting tomato.     

Molecular characterization and experimental host range of Euphorbia mosaic virus-Yucatan Peninsula, a begomovirus species in the Squash leaf curl virus clade

Euphorbia mosaic virus (EuMV), a tentative species within the genus Begomovirus, was isolated from Euphorbia heterophylla plants growing in the Yucatan Peninsula, Mexico. The complete bipartite genome was cloned from total DNA extracts and the nucleotide (nt) sequence was determined. The DNA-A sequence of the EuMV-Yucatan Peninsula (EuMV-YP) isolate shared 95% nt identity with the partially characterized type EuMV isolate from Puerto Rico. The EuMV-YP genome organization was like that of other New World, bipartite begomoviruses. The DNA-A component was 2613 nt in size, while the DNA-B component was 2602 nt long. The 165-nt common region (CR) sequence for the DNA-A and DNA-B components shared a lower than expected nt identity of 86%. The organization and iterons of the putative AC1 binding site of EuMV-YP were similar to those of begomoviruses in the Squash leaf curl virus (SLCV) clade. Characteristic disease symptoms were reproduced in E. heterophylla plants inoculated at the seedling stage using the cloned viral DNA-A and DNA-B components, confirming disease aetiology. Results of an experimental host-range study for EuMV-YP indicated that it infected at least five species in three plant families, including the Euphorbiaceae ( E. heterophylla ), Solanaceae ( Datura stramonium , pepper, tomato) and Fabaceae (bean). Phylogenetic analysis of the DNA-A and DNA-B components indicated that EuMV-YP is a New World begomovirus and that it is a new member of the SLCV clade.     

Dominance of a single clonal lineage in the Phytophthora infestans population from northern Shaanxi,China revealed by genetic and phenotypic diversity analysis

Late blight caused by Phytophthora infestans is the most serious disease of potato worldwide. To understand the P. infestans population structure in northern Shaanxi, an emerging potato production region in China, 125 single‐lesion isolates were randomly collected from farmers' fields in 2009 and characterized phenotypically and genotypically. A mating type assay showed that 94 isolates were A1 mating type. Virulence determination of selected isolates on a set of differential potato lines containing R1 to R11, respectively, showed the presence of two pathotypes, of which the pathotype lacking avirulence genes Avr3, Avr4 and Avr10 was dominant. Isolates lacking all avirulence factors Avr1 to Avr11 were detected but at lower frequency (13·6%). Analysis for mtDNA haplotype showed all 61 examined isolates were IIa. A total of seven multilocus genotypes were distinguished among 125 isolates, as determined with seven polymorphic microsatellite markers. The genotype SG‐1 was dominant in the population with a frequency of 75·2% and was present throughout the region. Analysis of the phenotypic and genotypic structures of P. infestans populations indicated strict clonal reproduction of the pathogen and suggested that sexual reproduction probably does not occur. Potential implications for disease management are discussed.     

Comparative analysis of pathogenic and nonpathogenic Fusarium oxysporum populations associated with banana on a farm in Minas Gerais,Brazil

Fusarium wilt is one of the most devastating diseases on banana. The causal agent, Fusarium oxysporum f. sp. cubense (Foc) is genetically diverse and its origin and virulence are poorly understood. In this study, pathogenic Foc isolates and nonpathogenic F. oxysporum isolates from Minas Gerais in Brazil were compared using EF‐1α and IGS sequences. This allowed the examination of the origin and evolutionary potential of Foc in a country outside the region of origin of the banana plant. Two different sequence types were found among Foc isolates. One appeared to be of local origin because it was identical to the sequence type of the largest group of nonpathogenic isolates. To explore if the ‘local’ Foc isolates had acquired pathogenicity either independently through coevolution with the host, or through horizontal gene transfer (HGT) of pathogenicity genes from other, probably introduced, Foc isolates, the presence and sequence of putative SIX effector genes were analysed. Homologues of SIX1, SIX3 and SIX8 were found. SIX1 sequences were identical and exclusively found in all pathogenic isolates, while variable ratios of sequences of multicopy gene SIX8 were found among nonpathogenic and different pathogenic isolates. This observation supports the HGT hypothesis. Horizontal transfer of genes between isolates of F. oxysporum has important implications for the development of reliable diagnostic tools and effective control measures. Full genome sequencing is required to confirm HGT and to further unravel the virulence mechanisms of forma specialis cubense.