Phenotypic and genetic characterization of Erwinia carotovora from mulberry (Morus spp.)

Phenotypic and genetic characteristics of nine bacterial strains isolated from mulberry ( Morus spp.), which were originally described as Erwinia carotovora ssp. carotovora (Ecc), were investigated. Based on the results of biochemical tests, these bacterial strains were divided into two different types, type 1 and type 2. Two strains of type 1 were similar to Ecc, whereas seven strains of type 2 were distinct from Ecc. A polyphasic study that included serological assay, specific PCR assay for E. carotovora ssp. atroseptica (Eca), PCR-RFLP of a pectate lyase ( pel ) gene and RAPD-PCR was performed on the type 2 strains, and the data were compared with those of related E. carotovora subspecies. The results of serological and specific PCR assays for Eca showed that the type 2 strains were distinct from Eca. In RFLP analysis of the pel gene using Sau 3AI, the type 2 strains showed a unique RFLP pattern. On the basis of RAPD analysis, similarity of RAPD patterns within the type 2 strains was very high. A unique RAPD fragment was isolated from the type 2 strains and used as a probe for Southern hybridization. This probe hybridized only with PCR products from the type 2 strains. Based on phenotypic, serological and genetic characteristics, the type 2 strains isolated from mulberry may belong to a distinct E. carotovora subspecies other than Eca or Ecc.     

Occurrence of a new microsporidium in the skeletal muscle of the flying fish Cypselurus pinnatibarbatus japonicus (Exocoetidae) from Yakushima, Japan

A new microsporidium was observed in the flying fish Cypselurus pinnatibarbatus japonicus (Franz) (Exocoetidae) from Yakushima, Japan. Visual examination revealed the microsporidium to form white elongate nodules in the host's trunk muscle. Monomorphic spores were ovoid to pyriform in shape, with average dimensions of 4.1 x 2.2 microm and possessing a polar tube describing 13-15 coils. Histological observations showed that each parasite focus of infection was encapsulated by a host-produced fibrous membrane. The presence of sporophorous vesicles was not clearly determined. Ribosomal DNA sequence analyses showed the microsporidium to be discrete from other known fish muscle-infecting species and to be most closely related to a clade comprising the Pleistophoridae and Glugea spp. The parasite is provisionally placed as Microsporidium cypselurus sp. n.     

Synthesis and characterization of synthetic analogs of cinnacidin, a novel phytotoxin from Nectria sp

BACKGROUND: The novel natural product cinnacidin was isolated from a fungal fermentation extract of Nectria sp. DA060097. The compound was found to contain a cyclopentalenone ring system with an isoleucine subunit linked through an amide bond. Initial biological characterization of cinnacidin suggested promising herbicidal activity. RESULTS: Two synthetic analogs, (2S,3S)-2-[(3RS,3aSR,6aRS)-3-methoxy-4-oxo-3,3a,4,5,6,6a-hexahydropentalen-1-ylcarbamoyl]-3-methylvaleric acid and benzyl (2S,3S)-2-[(3RS,3aSR,6aRS)-3-methoxy-4-oxo-3,3a,4, 5,6,6a-hexahydropentalen-1-ylcarbamoyl]-3-methylvalerate, were prepared for further characterization, and their herbicidal activities were compared with that of cinnacidin. CONCLUSIONS: The synthetic compounds were highly phytotoxic on a range of weeds. Based on the symptoms in treated plants, the mode of action of these compounds is suggested to be similar to that of coronatine and jasmonic acid. Coronatine was more active against warm-season grasses, while the cinnacidin benzyl ester analog was more effective on cool-season grasses. In a seedling growth bioassay conducted on bentgrass, the cinnacidin analog was equivalent in activity to coronatine.     

Biological and molecular characterization of a novel carmovirus isolated from angelonia

ABSTRACT A new carmovirus was isolated from Angelonia plants (Angelonia angustifolia), with flower break and mild foliar symptoms, grown in the United States and Israel. The virus, for which the name Angelonia flower break virus (AnFBV) is proposed, has isometric particles, approximately 30 nm in diameter. The experimental host range was limited to Nicotiana species, Schizanthus pinnatus, Myosotis sylvatica, Phlox drummondii, and Digitalis purpurea. Virions were isolated from systemically infected N. benthamiana leaves, and directly from naturally infected Angelonia leaves, using typical carmovirus protocols. Koch's postulates were completed by mechanical inoculation of uninfected Angelonia seedlings with purified virions. Isometric particles were observed in leaf dips and virion preparations from both Angelonia and N. benthamiana, and in thin sections of Angelonia flower tissue by electron microscopy. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis of dissociated purified virus preparations, a major protein component with a molecular mass of 38 kDa was observed. Virion preparations were used to produce virus-specific polyclonal antisera in both Israel and the United States. The antisera did not react with Pelargonium flower break virus (PFBV), Carnation mottle virus (CarMV), or Saguaro cactus virus (SgCV) by either enzyme-linked immunosorbent assay or immunoblotting. In reciprocal tests, antisera against PFBV, CarMV, and SgCV reacted only with the homologous viruses. The complete nucleotide sequence of a Florida isolate of AnFBV and the coat protein (CP) gene sequences of Israeli and Maryland isolates were determined. The genomic RNA is 3,964 nucleotides and contains four open reading frames arranged in a manner typical of carmoviruses. The AnFBV CP is most closely related to PFBV, whereas the AnFBV replicase is most closely related to PFBV, CarMV, and SgCV. Particle morphology, serological properties, genome organization, and phylogenetic analysis are all consistent with assignment of AnFBV to the genus Carmovirus.     

Phylogenetic analyses of plant-growth-promoting rhizobacteria isolated from tomato,lettuce, and Japanese pepper plants in Hyogo Prefecture,Japan

Approximately 30,000 fluorescent bacterial strains isolated from tomato, lettuce, eggplant, Chinese cabbage, and Japanese pepper plants at seven different locations in Hyogo Prefecture, were screened for plant-growth-promoting (PGP) activity to induce disease resistance against bacterial wilt in tomato. The 37 strains that had higher PGP activity were subjected to molecular phylogenetic analyses using the sequences of the 16S rRNA, gyrB and rpoD genes. Most of the strains were identified as Pseudomonas fluorescens or its close relative, P. putida, while a few strains were grouped with more distantly related bacterial species such as Enterobacter and Stenotrophomonas. The phylogenetic relationships among tomato and lettuce isolates mostly coincided with the source locality and host plants, with a few exceptions. In contrast, isolates from Japanese pepper plants did not form their own cluster but represented several different bacterial species.     

A note on the carpenter moth Salagena sp., a new pest of cashew in Zambia

Abstract

Salagena sp. (Lepidoptera: Metarbelidae) is reported for the first time as a pest of cashew. The larva feeds on the bark and bores galleries in the wood, covering the point of attack with a typical mass of frass and silk. The attack is widespread on most adult cashew trees In the western province of Zambia, severely reducing their vigour and opening the way to secondary invaders leading to progressive decline of the trees. Seasonal cycle and preliminary control measures including removal of the grubs, pruning, spraying and swabbing with insecticide, are briefly described.     

A new Monogenean species, Bicotyle lucknowensis sp. n. (Monogenea: Heteraxinidae), from the Indian fish, Gudusia chapra (Ham.)

A new monogenean species, Bicotyle lucknowensis sp. n. of the family Heteraxinidae, has been described from the fish, Gudusia chapra (Ham.), from Lucknow, India. The new species is characterized by the number and shape of cirral spines, presence of an unarmed vagina and details of copulatory complex and haptoral armature.     

Granulinema gen. n. a new dracunculoid genus with two new species (G. carcharhini sp. n. and G. simile sp. n.) from the bull shark, Carcharhinus leucas (Valenciennes), from Louisiana, USA

Two new species of dracunculoid nematodes, G. carcharhini sp. n. and G. simile sp. n., representing a new genus Granulinema gen. n. (Dracunculoidea: Micropleuridae) are described from the bull shark, Carcharhinus leucas, from Louisiana (Lake Borgne), USA; the site of their localization in the host is unknown (probably abdominal cavity). The nematodes of both species were found in tissue juice and only males, juvenile females and body fragments of more advanced but nongravid females were obtained. Granulinema gen. n. differs from Micropleura, the only other genus in the family Micropleuridae, mainly by the presence of marked, dark excretory corpuscles in lateral excretory canals, pointed tail in females, greater number (6) of postanal pairs of caudal papillae in males, and by the presence of conspicuous transverse cuticular ornamentations on the body surface of mature females. The two new species can be easily distinguished from each other by the length of their spicules (0.78-0.90 mm in G. carcharhini sp. n. and 0.20-0.36 mm in G. simile sp. n.); moreover, there are two pairs of preanal papillae in the male of G. carcharhini sp. n., while there are three pairs in G. simile sp. n.     

水稻黑条矮缩病毒江苏玉米分离物全基因组序列及进化分析

 利用RT-PCR从江苏玉米上克隆水稻黑条矮缩病毒(RBSDV)的全基因组,其基因组10个片段核苷酸数目和蛋白编码情况与已报道的RBSDV分离物基本一致。 基因组的序列一致性分析和系统进化树表明,RBSDV基因组片段间存在频繁的RNA重排现象;江苏玉米分离物(JS)与湖北玉米分离物(HuB)和河北小麦分离物(HeB)的整体亲缘关系较近,而与安徽分离物(AH-MX8)的整体亲缘关系最远。综合本研究及前人对S8 和S10的研究,RBSDV基因组不同片段的进化中RNA重组的作用不同: S1、S2和S4中没有RNA重组;S3、S5、S6、S8和S10中存在低频率重组;S7 和S9存在较高频率重组,其中S7的高频率重组尤为显著。     

Isolation,characterization, and properties of factor F1 from mitochondrial preparations of housefly (Musca domestica L.) thorax

A water-soluble Mg²⁺-dependent ATPase (coupling factor F₁) was isolated from the mitochondria of housefly thorax. It comprised about 14% of the proteins from a crude preparation. The F₁ preparation was nearly homogeneous as assessed by gel electrophoresis, isoelectric focusing, and electron microscopy. It was composed of five subunits with the following apparent molecular weights: α, 68,000; β, 61,000; γ, 38,000; δ, 27,000; and ?, 17,500. The isoelectric pH (pI) of this protein was 7.3. F₁ had a pH optimum of 8.2 and a temperature optimum between 37 and 45°C. The enzyme was fairly stable at 25°C. Nearly complete loss of activity was noticed at 0°C, while at 0 or 25°C, glycerol (20%) partially stabilized the enzyme activity against such inactivation. The K_m value of the enzyme with respect to ATP was 0.4 mM. The activity was stimulated by low concentrations of 2,4-dinitrophenol. The enzyme was inhibited by azide, p-hydroxymercuribenzoate, and guanidine hydrochloride. Oligomycin and the pesticides pyrethrin, cyhexatin, and DDT have no effect on the enzyme activity. However, all of these chemicals inhibited intact Mg²⁺- ATPase. The results are discussed in the light of differential responses of soluble and intact ATPase to these pesticides.     

乌蔹莓在果桑茶园的发生规律及化学防除

乌蔹莓[Cayratia japonica(Thunb.)Gagnep.]又名五爪龙,葡萄科乌蔹莓属多年生草质藤本植物。乌蔹莓是果桑茶园普遍发生的恶性杂草,单株生物量大,种子繁殖,蔓延快,如不及时防除,危害极其严重。现将近年来所作田间调查、定点观察及防除试验总结如下。1分布及危害分别在江苏省仪征市青山(代表丘陵山区黄土地段)、胥浦(代表圩区堤坡沙土地段)、夏州(代表圩区平原湿润沙壤土地段)3个镇,取有代表性果桑茶园3块,每块田调查10个样方(每样方1 m2),计算密度(株数/m2)、盖度(垂直投影覆盖率)、频度(样方中出现次数)。调查时间为5月上旬,结果见表1。由…     

Insecticide resistance management of permethrin and acephate against the cowpea curculio,Chalcodermus aeneus Boheman (Coleoptera: Curculionidae), a pest of the southern pea,Vigna unguiculata (L.) Walp.

To evaluate strategies for management of potential resistance of the cowpea curculio, Chalcodermus aeneus Boheman, to sprayer‐applied permethrin and acephate, five management regimes were evaluated on early, midseason and late sequential plantings of southern peas, Vigna unguiculata (L.) Walp. during 1991–1993. Management regimes were: (1) consecutive permethrin (0.11 kg a.i./ha early‐midseason‐late); (2) consecutive acephate (1.1 kg a.i.ha early‐midseason‐late); (3) consecutive mixture (0.055kg a.i./ha permethrin and 0.55kg a.i./ha acephate early‐midseason‐late); (4) alternation (0.11 kg a.i./ha permethrin early and 1.1 kg a.i./ha acephate midseason and late plantings); and (5) an untreated check early, midseason and late. The effect of management regime on the toxicities (LC₅₀) of permethrin and acephate was determined by topical application to next generation adult cowpea curculios reared from infested southern pea pods harvested from untreated plants in the management fields. In the field, infestation varied more by planting date than by management regime. Toxicity of permethrin to adults selected in the consecutive mixture regime was significantly lower in 1991 and 1992 late season and in 1993 midseason than in early season for the respective years. There was a seasonal decrease in the laboratory toxicity of acephate to adults with all management regimes.     

Some diagnostic, biologic and morphologic characteristics of Francisella tularensis strains isolated from the ticks Ixodes ricinus (L.) in the Prague agglomeration

Four infectious agents were isolated from the ticks Ixodes ricinus (L.) collected in the recreational area and park-forest of Prague. On the basis of cultivation, staining, biochemical, serologic properties, pathogenicity for animals and histological tests they were identified as Francisella tularensis with the following features: they are short, gram-negative rods of approximate dimensions of 0.3 X 0.8 micron, growing in enriched media after 3-4 day incubation at 37 degrees C. They form small circular, at first transparent, later greyish turbid colonies with regular rims. They are little active biochemically. They are susceptible to streptomycin and some broad spectrum antibiotics. They react positively with tularemic serum, but in lower titres than those in which this serum reacts with standard antigen. The microbes are highly pathogenic for mice, guinea-pigs, young rats, in which a massive bacteriemia occurs before death, but they do not kill rabbits. They multiply well in chick embryo, but do not grow in cell or tissue cultures. The most important histologic changes were observed in liver and spleen of mice. No pathologic changes were found in brain, lungs, heart, kidneys. Necroses were found in liver and in their marginal zones the microbes were present. Conspicuous were changes in numerous hepatocytes which became enlarged due to microbial multiplication and finally transformed into "sacs" packed with microbes. Histological and electronoptical examination showed that these are intracellular parasites fringed with a light lytic zone. Discussed is the problem to what extent the properties of the isolated strains are typical of F. tularensis as well as the importance of their detection from the aspect of epidemiology and differential diagnostics.     

麦麸饲料中细菌对家蝇幼虫生长发育的影响

从饲料用麦麸中分离出的12种细菌均能支持无菌家蝇Musca domesticaL.幼虫在胰化酪蛋白大豆卵黄琼脂(Trypticase Soy Egg Yolk Agar,TSEYA)培养基中完成整个生长发育过程。1)幼虫在接种香味类香味菌Myroides odoratimimus的TSEYA培养基中生长时间最短,仅需97.61±1.14h;2)幼虫在接种醋酸钙不动杆菌Acinetobacter calcoaceticus的TSEYA培养基中的化蛹率可达到86.81%;3)从接种嗜水汽单胞菌Aeromonas hydrophila的TSEYA培养基中得到的蝇蛹重量最高,达到20.15±0.23mg/个;4)除铜绿假单胞菌Pseudomonas aeruginosa饲养的家蝇羽化率较低(60.87%)外,其余各种细菌饲养的羽化率在84.33%～97.47%之间。此外,枯草芽孢杆菌Bacillussubtilis、香味类香味菌、聚团肠杆菌Enterobacter agglomerans以及成团肠杆菌Pantoea agglomerans可作为单一营养来源支持幼虫完成整个生长发育过程。对枯草芽孢杆菌、金黄色葡萄球菌S...     

Cloning and characterization of an NBS-LRR resistance gene from peanuts (Arachis hypogaea L.)

The nucleotide-binding site (NBS)-leucine-rich repeat (LRR) gene family accounts for the largest number of known disease resistance genes and is one of the largest gene families in plant genomes. In this study, resistance gene analogs (RGAs) were isolated from peanuts based on the NBS domain. A full-length cDNA, PnAG₃, was obtained by rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the length of PnAG₃ was 1882 bp, which included a complete open reading frame of 1335 bp that encoded for the PnAG₃ protein composed of 444 amino acids. Multiple analyses showed that this protein had homology with known resistance proteins, the highest being 48.01% with a resistance protein from Arachis cardenasii. The polypeptide has a typical non-TIR-NBS-LRR gene structure. Real-time fluorescence quantitative PCR analysis showed that after Aspergillus flavus infection, expression of the PnAG₃ gene in J11 (A. flavus-resistant species) increased by 16.68, 11.16 and 25.96 in the seed coat, kernel and pericarp, respectively. However, it only increased 2–3 times in JH1012 (A. flavus-sensitive species). Cloning of the putative resistance gene from peanut provides a basis for studying the structure and function of peanut disease resistance-related genes and disease resistance genetic breeding in peanuts.