<Emphasis Type="Italic">Fusarium mangiferae</Emphasis> associated with mango malformation in the Sultanate of Oman

Mango malformation, caused by Fusarium mangiferae, represents the most important floral disease of mango. The first symptoms of this disease were noticed in the beginning of 2005 in plantations at Sohar in the Sultanate of Oman. The affected inflorescences were abnormally enlarged and branched with heavy and dried-out panicles. Based on morphology and DNA-sequence data for the genes encoding translation elongation factor 1α and β-tubulin, the pathogen associated with these symptoms was identified as F. mangiferae.     

Aster yellows subgroup (Candidatus Phytoplasma sp. AY 16S-group,AY-sg) phytoplasma associated with porcelain vine showing witches' broom symptoms in South Korea

 This is the first report of a phytoplasma in porcelain vine [Ampelopsis brevipedunculata var. heterophylla (Thunb.) Hara.] with severe witches' broom symptoms in Korea. On the basis of the polymerase chain reaction-amplified ribosomal DNA, the phytoplasma infecting porcelain vine was classified as a member of the aster yellows subgroup. Received: October 21, 2002 / Accepted: December 20, 2002     

Role of the α subunit of heterotrimeric G-protein in probenazole-inducing defense signaling in rice

 Certain cellular responses to stresses and stimuli are regulated by a G-protein-mediated signaling pathway. A rice dwarf mutant that is defective in the α subunit of the heterotrimeric G-protein was found to be fully protected from blast fungus by the plant activator probenazole (PBZ) despite the 1-day delay in induction of the PR-10 gene PBZ1 by PBZ. These results suggest that the signaling pathway for protection by PBZ is not via the G-protein, although G-protein is involved in the induction of PBZ1 by PBZ. Received: March 27, 2002 / Accepted: August 20, 2002     

Characterization of a phytoplasma associated with witches' broom disease of potatoes in Korea

 Symptoms of witches' broom disease caused by phytoplasma, including general stunting and yellowing, were observed in potatoes (Solanum tuberosum L.) in a storehouse on Jeju Island, Korea in 1998. Based on sequence analysis of DNA products from polymerase chain reaction (PCR)-amplified 16S ribosomal DNA and 16S–23S spacer region using universal phytoplasma primers, the phytoplasma associated with potato witches' broom disease (PWB) was identified as a member of 16S-group VIII. It was most closely related to elm AH phytoplasma (99.7% similarity, accession no. AF268895), which is in the clover proliferation (CP) subgroup. This report is the first from the East Asian continent of a plant pathogenic phytoplasma belonging to the CP subgroup and includes the nucleotide sequence of most of the potato phytoplasma 16S rDNA. Received: May 1, 2002 / Accepted: July 1, 2002     

Homolog of FlhDC, a master regulator for flagellum synthesis: required for pathogenicity in Erwinia carotovora subsp. carotovora

 Erwinia carotovora subsp. carotovora (Ecc) is a causal agent of soft-rot diseases in a wide variety of plants. Here, we have isolated nonmotile mutants in Ecc by in vivo insertional mutagenesis using a transposon Tn5. The sequence disrupted by the Tn5 insertion in YMU1 and YMU5 mutants was highly homologous to that of flhC and flhD genes, respectively. They are involved in the initiation of the expression of flagellum-related genes in many gram-negative bacteria such as Escherichia coli and Salmonella. With electron microscopy, the flhC and the flhD homolog mutants were shown to be aflagellate. Furthermore, the virulence of these mutants was greatly reduced in Chinese cabbage and potato compared to that of the parental strain. These results suggest that flagellar formation is required for the pathogenicity of Ecc. Received: November 5, 2002 / Accepted: December 2, 2002 Acknowledgments This research was supported in part by Grant-in-Aid (12052210) and by a grant from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (13073).     

d -Alanine-d -alanine ligase gene ( ddl ) of Erwinia chrysanthemi strain EC16 II: analysis of pectate lyase regulation using ddl ? mutant

 Erwinia chrysanthemi (Ech) triggers soft rot disease mainly by secreting pectate lyase (Pel), which is regulated in a complex manner by many regulatory genes. In a previous study, we used a gene dosage method to show that the ddl gene, which encodes d-alanine-D-alanine ligase, reduced Pel production and tissue maceration by Ech strain EC16n. In this study, the ddl ⁻ marker-exchanged mutant was shown to overcome the long growth lag caused by various salts in the growth medium and to increase Pel production over that by EC16n, especially in a medium containing magnesium salts. Thus, ddl seems to regulate Pel production in a negative manner. Because the profiles of a gel shift assay using the pelE promoter region as the target DNA with crude extracts of EC16n and ddl ⁻ mutant were distinguishable, Ddl is thought to affect the binding of other regulatory proteins. Expression of the ddl gene was induced in the medium containing a low-molecular-weight fraction of potato extract, but it was reduced in that containing both polygalacturonic acid (PGA) and the fraction. The repression of ddl expression by PGA should contribute in part to the in planta hyperinduction of Pel. Received: May 15, 2002 / Accepted: June 20, 2002     

Effect of rapid and gradual increase of osmotic stress on survival of entomopathogenic nematodes

The effect of rapid and gradual exposure of entomopathogenic nematodes to osmotic stresses on the induction of a dormant state was determined with the nematodeSteinernema feltiae IS-6 infective juveniles (IJs). Rapid exposure of nematodes to glycerol at concentrations of 24% and 28% (w/w) caused the nematodes to enter a dormant state which was characterized by shrinking and impeded motility of all nematodes within 8 h. However, pre-exposure to gradually increasing glycerol concentrations of 5%, 10% and 18% at 4-h intervals resulted in dormancy after 4 h exposure to 24% glycerol. The total time of exposure to glycerol solution was 16 h in gradual osmotic stress. For nematodes exposed to 24% glycerol solution either rapidly or gradually, recovery occurred after 40 min in distilled water. Infectivity of osmotically stressedS. feltiae IJs was evaluated by two criteria, insect mortality and invasion rate. The assays indicated that infectivity of nematodes desiccated by rapid and gradual osmotic stresses was similar to that of fresh nematodes. Rapid exposure ofS. carpocapsae ‘All’,S. riobravis ‘Texas’,S. glaseri ‘NI’ andHeterorhabditis bacteriophora HP88 IJs to the 24% glycerol solution resulted in dormancy within 8 h. These treatments caused mortality of 48.4% and 11.7% amongS. glaseri Nl andH. bacteriophora HP88 IJs, respectively. Similar effects were observed when these nematode species were exposed to increasing osmotic stress of 5%, 10% and 18% at 6-h intervals. Under these same conditions, mortality ofH. bacteriophora HP88 andS. glaseri Nl IJs was 27.5% and 61.8%, respectively. http://www.phytoparasitica.org posting Sept. 29, 2004.     

Diverse <Emphasis Type="Italic">Fusarium solani</Emphasis> isolates colonise agricultural environments in Ethiopia

Fusarium solani is a fungal pathogen that infects many different genera of plants. It represents one of the two Fusarium spp. commonly isolated from agricultural soils and plant tissues in Ethiopia. To determine the diversity of F. solani in Ethiopia, we studied 43 isolates using Amplified Fragment Length Polymorphism (AFLP) and nucleotide sequences of the Translation Elongation Factor 1α (TEF-1α) and β-tubulin genes. TEF-1α sequences from GenBank, representing previously described species and clades of the F. solani-Haematonectria haematococca complex, were also included for comparative purposes. Phylogenetic analyses of the TEF-1α data separated the isolates into three groups corresponding with the three previously described clades (Clades 1–3) for this fungus. The Ethiopian isolates aggregated into one group corresponding to Clade 3. TEF-1α, β-tubulin and AFLPs further separated the Ethiopian isolates into a number of clusters and apparently novel phylogenetic lineages. Although the biological and ecological significance of these lineages and clusters is unclear, our data show that the Ethiopian agricultural environment is rich in species and lineages of the F. solani-H. haematococca complex.     

Sensitivity of Fusarium moniliforme Isolates to Ipconazole

To estimate the sensitivity of Fusarium moniliforme to ipconazole, a sterol biosynthesis inhibitor (SBI), minimum inhibitory concentrations (MIC) were determined for isolates which were collected before the launch of ipconazole as a rice seed disinfectant. Research institutes from various prefectures in Japan supplied 211 isolates (group I) from their collections, and 84 isolates (group II) were isolated from rice paddy fields in Iwaki, Fukushima Prefecture. In group I, the MIC ranged from 0.10 to 6.25 μg/ml with a peak at 0.39 μg/ml. In group II, MIC values had the same range as group I, but the main peak was at 0.20 μg/ml. Ipconazole sensitivity did not differ significantly among groups I and II. Though the ranges of MIC values for ipconazole, pefurazoate and triflumizole were different in 60 isolates randomly chosen from group I, positive correlations were observed in their sensitivities to SBIs, suggesting a common mechanism in F. moniliforme for lowering sensitivities to SBIs. Among the 14 isolates tested, isolates with MIC values lower than 0.78 μg/ml for ipconazole were pathogenic to rice seedlings, and all the isolates with MIC values higher than or equal to 1.56 μg/ml were not pathogenic in the nursery test. Good protection against isolates causing “Bakanae” disease was obtained by dipping seeds for 24 hr in ipconazole. The pathogenic isolates can be controlled by the seed treatment with the practical dosage of ipconazole because of the adequate margin between the highest MIC value for the pathogenic isolates and the treatment concentration. In addition, the low or lack of pathogenicity of the isolates less sensitive to ipconazole may also contribute to the stable efficacy of ipconazole. Received 8 November 1999/ Accepted in revised form 18 April 2000     

Nucleotide sequence of the coat protein gene of Mirafiori lettuce virus

 The coat protein (CP) gene of Mirafiori lettuce virus (MiLV), a tentative member of the genus Ophiovirus was isolated and sequenced. The established sequence consists of 1514 nucleotides including one open reading frame (ORF) with 1311 nucleotides that encodes 437 amino acids with a relative molecular mass 48 543. When the ORF was expressed in Escherichia coli, the obtained protein was confirmed as CP by Western blotting using an antiserum against MiLV. Database searches showed that the CP gene of MiLV has a sequence similar to that of Citrus psorosis virus (CPsV), the type species of the genus Ophiovirus. The comparison between MiLV and CPsV CP genes revealed that the identities of the nucleotide and amino acid sequences were 46.5% and 30.9%, respectively. Received: July 29, 2002 / Accepted: October 2, 2002     

新疆碳排放估算及其特征分析

由于二氧化碳有较长的寿命年限及超高排放量,已成为温室效应的标志性气体。依据IPCC（2006）自上而下一阶估算的参考方法,计算得出新疆2003-2011年能源消费产生的二氧化碳量,结果表明：该地区二氧化碳排放量逐年迅速增长,主要表现为固体燃料燃烧产生的二氧化碳的增长,且排放量较高的重点行业主要以化工、电力、钢铁、水泥为代表的高耗能行业。结合新疆能源、经济及环境现状,提出了建立市场准入机制、碳交易机制、优化产业结构及提高能源利用效率等一系列促进新疆低碳经济发展的相关建议。     

Morphological and molecular characterization for a Japanese isolate of tomato powdery mildew Oidium neolycopersici and its host range

 A single conidium of tomato powdery mildew was isolated from heavily infected leaves of tomato (cv. Moneymaker) grown in the greenhouse of Kinki University, Nara Prefecture, Japan. It was successively multiplied so the morphological and taxonomic characteristics of the pathogen and its host range under high humidity conditions could be analyzed. The isolate KTP-01 of the tomato powdery mildew optimally developed infection structures at 25°C under continuous illumination of 3500 lx. More than 90% of the conidia germinated and developed moderately lobed appressoria. After forming haustoria, the pathogen elongated secondary hyphae from both appressoria and conidia. The hyphae attached to leaf surfaces by several pairs of appressoria and produced conidiophores with noncatenated conidia. In addition to its morphological similarity to Oidium neolycopersici, the phylogenetic analysis (based on the sequence of internal transcribed spacer regions of rDNA) revealed that KTP-01 could be classified into the same cluster group as O. neolycopersici. In host range studies, KTP-01 produced abundant conidia on the foliage of all tomato cultivars tested and tobacco (Nicotiana tabacum), and it developed faint colonies accompanied by necrosis on leaves of potato (Solanum tuberosum), red pepper (Capsicum annuum), petunia (Petunia × hybrida), and eggplant (S. melongena). The pathogen did not infect other plant species including Cucurubitaceae plants, which have been reported to be susceptible to some foreign isolates. Thus, the present isolate of the tomato powdery mildew was assigned as O. neolycopersici, a pathotype different from foreign isolates of the pathogen. Received: December 5, 2002 / Accepted: December 26, 2002 Acknowledgments This work was supported in part by a Grant-in-Aid (12660050) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan. We express our deepest thanks to professor Dr. Y. Sato, Toyama Prefectural University, for his kind and valuable suggestion on taxonomic analysis of the powdery mildew pathogen described in the present study.     

Complete set of extrachromosomal DNAs from three pathogenic lines of onion yellows phytoplasma and use of PCR to differentiate each line

 Two lines of onion yellows phytoplasma with reduced pathogenicity have been isolated from the original wild-type line (OY-W). One is a line with mild symptoms (OY-M) and the other is a non-insect-transmissible line, also with mild symptoms (OY-NIM). We previously reported heterogeneity in extrachromosomal DNA (EC-DNA) species in these lines. In this report, another EC-DNA, EcOYNIM, from OY-NIM was cloned and sequenced, providing a complete set of EC-DNAs from the three OY lines. To monitor each phytoplasma in synergism or cross-protection experiments, a pair of polymerase chain reaction (PCR) primers that universally amplify a portion of the EC-DNAs that are characteristic of each line was designed. Using this primer set, a line-specific fragment was amplified from the total DNA of each plant inoculated with one or more phytoplasma lines. The PCR product sizes differ for each phytoplasma line, so the lines can be distinguished even in plants infected with multiple lines. Because EC-DNAs are more abundant than chromosomal genes in phytoplasma cells, this primer set will be valuable for detecting and discriminating these phytoplasma lines and for analyzing their interaction. Received: October 21, 2002 / Accepted: January 8, 2003 RID="*" ID="*" The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AB097150 Acknowledgments This work was supported partly by Grants-in-Aid of Scientific Research from the Japan Society for the Promotion of Science (JSPS) (09460155 and 13306004), a Grant-in-Aid of Scientific Research on Priority Areas (C) “Genome Biology” from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and the Program for the Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN) of the Bio-oriented Technology Research Advancement Institution.     

Distinguishable staining with neutral red for GFP-marked and GFP-nonmarked <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> strains simultaneously colonizing root surfaces

 Green fluorescent protein (GFP)-marked Fusarium oxysporum f. sp. melonis and nonmarked F. oxysporum f. sp. fragariae were stained with neutral red. The neutral red stained vacuoles of the fungi without disturbing GFP fluorescence in the cytoplasm. GFP-marked fungi showed fluorescent hyphae with dark-stained vacuoles, whereas nonmarked fungi were detected as nonfluorescent hyphae with dark-dotted vacuoles. Root colonization by these two fungi was monitored using this method. Microconidia attached similarly to the root surface and elongated vegetative hyphae. Only the pathogenic fungi invaded, causing necrosis at the inoculation site. Thus, the present method enabled us to track simultaneously the various formae speciales of F. oxysporum colonizing the root surface. Received: March 25, 2002 / Accepted: September 27, 2002     

Rapid detection of chitosanase activity in chitosanase gene-transformed strains of <Emphasis Type="Italic">Enterobacter cloacae</Emphasis> by lytic infection of specific bacteriophages

 In combination with lytic infection by virulent phages, a simple method for monitoring transgenic strains of Enterobacter cloacae was developed in this study. First, 15 strains of E. cloacae were used as indicator bacteria to isolate virulent phages with different host ranges. Of the phages isolated, five isolates (EcP-22, -35, -45, -55, and -70) were used to construct a set of virulent phages corresponding to all strains of E. cloacae. Using this phage set, a rhizosphere strain (KRM-055E) of E. cloacae was effectively screened from field soil. KRM-055E was transformed with a prokaryotic chitosanase gene csnSM1 and infected with the phage EcP-03, which can lyse the strain most effectively. The lysis of KRM-055E/csn occurred 2 h after inoculation, and the chitosanase activity was simply detected by dropping the lysate onto an agar plate containing glycol chitosan. The positive signal for chitosanase activity was detected in the 2-h lysates, and the signal intensity reached a maximum in the 5-h lysate. The present assay was simple, rapid, inexpensive, easy to perform, and applicable to another strains. Received: August 2, 2002 / Accepted: October 31, 2002 Acknowledgments This work was supported in part by a grant (no. 99L01205) from the “Research for the Future” program of the Japan Society for the Promotion of Science. We are grateful to Dr. M. Sato, National Institute of Agrobiological Science, Dr. H. Okamoto, Fukui Agricultural Experiment Station, and Dr. K. Tsuda, Kyoto Prefectural Institute of Agricultural Biotechnology, for kindly providing E. cloacae strains. We thank Dr. P. Park, Kobe University, for technical support with the electron microscopic observations.     

气候变化对台兰河年平均径流量的影响

以台兰河为研究对象,选用台兰水文站1957-2011年平均流量序列,采用SPSS统计软件对其55 a序列进行趋势分析。结果显示：台兰河径流与降水量的年内分配极不均匀,径流主要集中在夏秋两季,降水量集中在春夏两季,径流与降水量集中期主要分布在7月。识别和提取台兰河年平均流量时间序列的趋势函数QS(t),获得了相应的趋势回归模型;台兰河年平均流量序列存在1957-1981年、1981-1999年、1999-2011年3个时段呈递减→递增→递减变化态势;降水、气温变化是影响台兰河不同时段年平均流量变化的主要因素,年平均径流量变化趋势与气温、降水变化趋势一致。     

Occurrence of chrysanthemum virescence caused by “<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma aurantifolia” in Okinawa

In 1999, a disease of chrysanthemum [Dendranthema grandiflorum (Ramat.) Kitamura], characterized by virescence of flowers, occurred in Okinawa Prefecture. The causal agent was identified as “Candidatus Phytoplasma aurantifolia” based on 16S rDNA sequencing. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession number AB247462.