Identification of Mulberry Dwarf Phytoplasmas in the Genital Organs and Eggs of Leafhopper Hishimonoides sellatiformis

ABSTRACT The presence of mulberry dwarf (MD) phytoplasmas in organs of the inoculative vector insects Hishimonoides sellatiformis and Hishimonus sellatus was determined by means of electron microscopy (EM) and polymerase chain reaction (PCR) assays. Many MD phytoplasmas were detected in genital organs as well as in the intestines, salivary glands, brains, fat bodies, and thoracic ganglia of Hishimonoides sellatiformis, but only in the intestine and salivary glands of Hishimonus sellatus. Many phytoplasmas with characteristic morphology were observed via EM in ovaries, seminal receptacles, and testes, and they were further identified by PCR assays with group I-specific primers. In addition, the organisms were detected by direct or nested PCR assays in eggs (head pigmentation stage of embryos) laid on mulberry shoots by inoculative leafhoppers and in the newly hatched nymphs from these eggs. These findings indicate that transovarial transmission of MD phytoplasmas occurs in Hishimonoides sellatiformis.     

Evaluating environmental impacts of the Japanese beef cow–calf system by the life cycle assessment method

The objectives of this study were to evaluate the environmental impacts of a beef cow–calf system using a life cycle assessment (LCA) method and to investigate the effects of scenarios to reduce environmental impacts on the LCA results. The functional unit was defined as one marketed beef calf, and the processes associated with the cow–calf life cycle, such as feed production, feed transport, animal management, the biological activity of the animal and the treatment of cattle waste were included in the system boundary. The present results showed that the total contributions of one beef calf throughout its life cycle to global warming, acidification, eutrophication and energy consumption were 4550 kg of CO₂ equivalents, 40.1 kg of SO₂ equivalents, 7.0 kg of phosphate (PO₄) equivalents and 16.1 GJ, respectively. The contribution of each process to the total environmental impact in each environmental impact category showed a similar tendency to the contribution of each process in each environmental category reported in the case of the beef fattening system as a whole. The results from this analysis showed that shortening calving intervals by 1 month reduced environmental impacts by 5.7–5.8% in all the environmental impact categories examined in the current study, and increasing the number of calves per cow also reduced environmental impacts in all the categories, although the effects were smaller.     

An epidemiological survey of Chrysanthemum chlorotic mottle viroid in Akita Prefecture as a model region in Japan

In epidemiological surveys on Chrysanthemum chlorotic mottle viroid (CChMVd) in various chrysanthemums cultivated in Akita Prefecture, Japan, in 2002–2005, approximately 20% of cultivars harbored CChMVd, including more symptomatic types than nonsymptomatic. Large-flowered cultivars were less frequently infected than small-flowered and spray types. The number of CChMVd-infected chrysanthemums is increasing, and the disease is found throughout major chrysanthemum-producing districts. Chrysanthemums infected only with CChMVd, in general, had no noticeable symptoms of disease. Most of those dually infected with known viruses and/or viroid also had no symptoms characteristic of chlorotic mottle disease. The lack of noticeable symptoms in major Japanese cultivars may have resulted in the unnoticed spread of the viroid.     

Identification of Petunia hybrida cultivars that diurnally emit floral fragrances

In order to identify genetic resources for breeding fragrant petunias for use as bedding plants, volatile compounds released by day from the flowers of 40 commercial Petunia hybrida cultivars were analyzed using a solid-phase micro-extraction technique coupled with GC–MS. The three cultivars with solid deep-blue flowers that accumulate malvidin in corollas with high tissue pH were found to emit abundant iso-eugenol as the principal floral fragrance. Several other cultivars that emitted considerable amounts of methylbenzoate and/or benzylbenzoate from the flower were also identified. Association between the floral fragrance and the other floral traits such as floral anthocyanin composition and corolla-tissue pH was discussed.     

猪隐秘杆菌型出血性坏死性脾炎

六月龄去势公猪表现昏睡、食欲不振、站立困难。尸检发现脾脏边缘呈多重出血灶。从脾脏、肾脏、肌肉和肝脏中分离出革兰氏阳性杆菌,对分离株(TO16177)的16S rDNA基因序列比较分析发现,其可能与未发表过的隐秘杆菌属HJ57-14E菌株(登记号:gi18873551)(比较675bp个碱基,相似性达99.7%)为同一个属。脾脏组织切片的组织学检查发现呈广泛性坏死和炎症,其中革兰氏阳性杆菌显而易见。肝脏中可见多病灶的坏死斑。免疫组织化学检测发现,分离株与抗化脓性隐秘杆菌属(Arcanobacterium pyogenes)和内氏放线菌(Actinomyces naeslundii)的多克隆抗体具有交叉反应,且与后者的交叉反应更强烈。相似的反应也见于扁桃体的化脓灶中分离株,偶尔也见于脾脏和淋巴结中的分离株。本研究结果表明,这种未公开发布的隐秘杆菌属的细菌会引起生长肥育猪多器官功能衰竭,而后继发急性出血性坏死性脾炎。     

Mottle necrosis of sweet potato caused by <Emphasis Type="Italic">Pythium scleroteichum</Emphasis> in Japan and varietal difference in susceptibility to the disease

Severe mottle necrosis (Shirogusare-byo in Japanese) was found on mature tubers of sweet potato (Ipomoea batatas) in Ibaraki Prefecture in October 2004. The causal organism was identified as Pythium scleroteichum hitherto unknown in Japan. Sweet potato cultivar Purple Sweet Lord was more susceptible than cultivars Beniazuma, Benimasari, Koukei-14, and Tamayutaka to the pathogen at 25°C, while this difference in the susceptibility was not clear at 15°C.     

A Conjugative Plasmid Carrying the efe Gene for the Ethylene-Forming Enzyme Isolated from Pseudomonas syringae pv. glycinea

ABSTRACT Previous work suggested that the efe gene encoding the ethylene-forming enzyme was present in the plasmids of three pathovars of Pseudomonas syringae including glycinea, phaseolicola (kudzu strains), and cannabina. However, no direct evidence to support this assumption had been presented. In the current study, we isolated the conjugative plasmid harboring the efe gene (ethylene plasmid) designated pETH2 from P. syringae pv. glycinea MAFF301683. pETH2 was detected by Southern blot hybridization using the efe probe, marked with the transposon mini-Tn5-Km1, and transferred into P. syringae Ni27(n), which does not produce ethylene. The transconjugant Ni27(n) (pETH2) produced ethylene at a level similar to pv. glycinea MAFF301683. In addition, the plasmid designated pCOR2, which encodes coronatine biosynthesis genes, was detected in the same strain. Although the molecular size of the plasmid pCOR2 was not easily distinguishable from pETH2, pCOR2 transferred independently into Ni27(n) and the transconjugants produced coronatine. These findings suggested that the efe gene has been horizontally dispersed among pathovars of P. syringae by plasmid-mediated conjugation in nature.     

Diazepam potentiates the positive inotropic effects of histamine and forskolin in guinea-pig papillary muscles

There have been diverse reports on the effects of diazepam on cardiac contractility. The purpose of this study was to examine whether diazepam modifies the inotropic response elicited by histamine on an isolated guinea-pig papillary muscle. The responses of electrically driven papillary muscle to histamine and cyclic AMP-related inotropic agents were recorded in the absence and in the presence of diazepam. Histamine and forskolin, which directly stimulate adenylate cyclase, significantly increased the contractile force in the papillary muscle in a concentration-dependent manner. A histaminergic H₂-receptor antagonist, cimetidine, but not a H₁-receptor antagonist, diphenhydramine, at 10 μ M produced a rightward shift in the concentration-response curve for histamine. Diazepam (10 μ M ) shifted the concentration-response curve for histamine and forskolin to the left by 1.8 and 1.6 times, respectively. Neither a central type (fulmazenil) nor a peripheral type (PK11195) of benzodiazepine receptor antagonist modified the effect of diazepam on the histaminergic-evoked contraction. Phosphodiesterase blockade by 3-isobutyl-1-methylxanthine shifted the concentration-dependent curve for histamine to the left. A combination of 3-isobutyl-1-methylxanthine also produced a leftward shift of the curve. However, there was no significant difference between the 3-isobutyl-1-methylxanthine only group and the combination group. These results indicate that diazepam potentiates the positive inotropic effect produced by histamine, probably mediated via an increase in cyclic AMP levels induced by histamine.     

Intracellular recording of rat neuron activity at the medial preoptic area of the hypothalamus using triangular wave microelectrode oscillation

A triangular wave microelectrode oscillating apparatus was constructed to evaluate an intracellular recording of rat neuron activity in the medial preoptic area (POA) of the hypothalamus. In this apparatus, electrodes passed a current with a frequency of 1.0 to 1.8 kHz and a voltage of 2.2 to 3.2 V and produced micro-oscillation of the electrode tip. The electrode was inserted into a neuron of the rat POA in vivo. In vivo recording of the activity of the rat POA neuron was possible. By means of electrical stimulation of the median eminence arcuate of the hypothalamus, an intracellular recording of antidromic, orthodromic or non-responding neuron was also possible. As a result, various components of the action potential such as the resting, threshold and spike potentials, and depolarization and repolarization such as after-hyperpolarization and after-depolarization were observed. The resting potentials ranged from 45 to 90 mV, and POA neurons possessed action potentials of almost the same magnitude. Several problems, however, remain to be solved. In general, the time available for the intracellular recordings is too short. The cells survive only for 15 minutes at the longest and may die in only a few minutes. An improvement of the apparatus was mandatory.