Spread of potato virus Y in potato cultivars (Solanum tuberosum L.) with different levels of sensitivity

The aim of this work was to correlate the appearance of the symptoms, multiplication and spread of virus after mechanical inoculation of potato (Solanum tuberosum L.) cultivars showing different levels of susceptibility and sensitivity to Potato virus Y^NTN (PVY^NTN). The potato cultivars used were the resistant cultivar Sante and susceptible cultivars Igor, Pentland squire and Désirée. The spread of the virus PVY^NTN in infected plants was monitored using different methods: DAS-ELISA, tissue printing, immuno-serological electron microscopy and real-time PCR. In all three susceptible cultivars, the virus was detected in the inoculated leaves 4–5 days after inoculation. From there virus spread rapidly, first into the stem, then more or less simultaneously to the upper leaves and roots. Real-time PCR was shown to be very sensitive and enabled viral RNA to be detected in non-inoculated leaves of susceptible cultivar Igor earlier than other methods. Therefore, for exact studies of plant–virus interaction, a combination of methods which detect viruses on the basis of their different properties (coat protein, morphology or RNA) should be used to monitor the spread of viruses.     

Isolation of two strains of a new Tombusvirus (Havel river virus, HaRV) from surface waters in Germany

Two virus isolates from water samples — one from a small stream in South Western Germany and another one from the Havel river in North Eastern Germany c. 500 km away, proved to be strains, named S and H, respectively, of a new Tombusvirus for which the name Havel river virus (HaRV) had been suggested previously in a brief account. Immunoelectron microscopical decoration tests and sequence comparisons of the coat proteins indicated that the two HaRV strains are only distantly related to known Tombusviruses. The closest relationships were found to Cucumber necrosis virus. Nothing is known about their natural hosts. Because the S strain of HaRV was isolated in a woody area from a small stream close to its origin, they may be pathogens of trees or wild plants in such habitats.     

Footsteps from Insect Larvae Damage Leaf Surfaces and Initiate Rapid Responses

Plant resistance to insect herbivory involves gene expression in response to wounding and the detection of insect elicitors in oral secretions (Kessler and Baldwin, 2002, Ann. Rev. Plant/ Biol. 53: 299–328). However, crawling insect larvae stimulate the synthesis of 4-aminobutyrate within minutes and imprints of larval footsteps can be visualized within seconds through superoxide production or transient increases in chlorophyll fluorescence (Bown et al., 2002, Plant Physiol. 129: 1430–1434). Here cryo-scanning electron microscopy was used to demonstrate that larval feet, which are equipped with a perimeter row of hook-like crochets, damage leaf tissue and result in larval footprints. Staining for cell death shows that areas of wounding correspond to footsteps detected through increased chlorophyll fluorescence. Superoxide production in response to footsteps was inhibited by diphenyleneiodonium, an inhibitor of the plasma membrane NADPH oxidase enzyme. Inhibition of superoxide production, however, did not eliminate the detection of cell death. The results demonstrate that larval footsteps damage leaf tissue, and initiate rapid local responses which are not dependent on herbivory or oral secretions. It is proposed that superoxide production at the wound site prevents opportunistic pathogen infection.     

Histological and proteomics analysis of apple defense responses to the development of Colletotrichum gloeosporioides on leaves

Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. is an important fungal pathogen known to cause glomerella leaf spot (GLS). The objective of this study was to analyze the infection of C. gloeosporioides on leaves of apple cv. Fuji (resistant) and cv. Gala (susceptible) and apply proteomics techniques to study the apple defense responses 48 h after inoculation (h.a.i.). On both of cultivars, C. gloeosporioides started to germinate at 3 h.a.i. on adaxial surface and produced appressoria adhering to epidermal cell juxtapositions. Histological analysis showed more stratified parenchyma in leaves of cv. Fuji than cv. Gala associated with differences in the chemical composition of cell walls. Total and unique proteins expressed by cvs. Fuji and Gala at 3, 12, 24 and 48 h.a.i. were detected by comparative proteomes analysis. A total of 42 unique proteins expressed at 24 and 48 h.a.i. were identified by MALDI/TOF mass spectrometry, and most of these proteins were identified as directly involved in defense responses.     

In vivo confocal microscopy in the normal corneas of cats, dogs and birds

OBJECTIVE: To evaluate the applicability of in vivo confocal microscopy (IVCM) in veterinary ophthalmology and analyze the morphology of living, healthy cornea. ANIMALS EXAMINED: Thirty-seven dogs, 34 cats and five birds. PROCEDURE: Various corneal sublayers were visualized in the central region using an in vivo confocal corneal microscope (HRTII/RCM). RESULTS: An investigation method was developed and adapted for use on animals with varying skull forms and eye positions. Real-time images of the epithelial cells, the corneal stroma and the endothelial layer were obtained. The corneal stromal nerve trunks and the subepithelial and basal epithelial nerve plexus were visualized. In dogs, full corneal thickness (FCT) was 585 +/- 79 microm (mean +/- SD) and endothelial cell density (ECD) 3175 +/- 776 cells/mm(2) (mean +/- SD). In cats, FCT was 592 +/- 80 microm and ECD 2846 +/- 403 cells/mm(2). There were no significant differences between canine and feline FCT and ECD and no morphologic differences could be seen between dogs and cats. The bird images revealed a number of structural differences. CONCLUSION: Noninvasive IVCM allows accurate detection of corneal sublayers, corneal pachymetry, endothelial cell density and corneal innervation in various animal species. For clinical usage, patients must be under general anesthesia. The confocal images provided anatomic reference images of various healthy corneal structures in dogs, cats and birds.     

大银鱼卵细胞粘丝扫描电镜观察

用扫描电镜观察了大银鱼的第Ⅲ时相，第Ⅳ时相，第Ⅴ时相卵母细胞和受精卵的卵细胞粘丝的外部形态特征和它们的粘性，并讨论了卵细胞粘丝的功能。     

Association of aster yellows subgroup 16SrI-B phytoplasmas with a disease of Rehmannia glutinosa var. purpurea

A new phytoplasma disease of Rehmannia glutinosa var. purpurea was observed in the Czech Republic in 1998. Infected plants showing severely proliferating shoots, leaves reduced in size with vein clearing and chlorosis, shortened internodes and virescent petals died in advanced stages of the disease. Electron microscopy examination of the ultra-thin sections revealed the presence of numerous polymorphic bodies in phloem tissue of leaf midribs and petioles. The disease was successfully transmitted from infected plant via a dodder bridge into periwinkle ( Catharanthus roseus ). The phytoplasma aetiology of this disease was further confirmed by polymerase chain reaction (PCR) using universal primers R16F2/R16R2. Restriction fragment length polymorphism (RFLP) analysis of amplification products indicated the presence of aster yellows related phytoplasmas (16SrI-B) in naturally infected samples of R. glutinosa var . purpurea and in symptomatic periwinkle after dodder transmission of the agent. A comparison of the amplified sequence with 17 sequences available in the GenBank confirmed the classification of the phytoplasma in the subgroup 16SrI-B. This is the first report of natural occurrence of phytoplasma-associated disease in R. glutinosa var. purpurea.     

福建果树根部的芒果半轮线虫记述

 芒果半轮线虫(Hemicriconemoides mangiferae Siddiqi,1961)在福建省一些果园发生。寄主有龙眼(Euphoria longana)、荔枝(Litchi chinensis)、芒果(Mangifera indica)、橄榄(Canarium album)、枇杷(Eriobotrya japonica)和葡萄(Vitis vinifera)等果树。雌虫唇部正面、环纹和阴门,雄虫侧带以及幼虫角质膜刺突等细微结构用扫描电子显微镜观察。     

Leek Proliferation: A New Phytoplasma Disease in the Czech Republic and Italy

During the summer 1996, twelve of twenty-eight leek plants located in a garden near eské Budjovice, South Bohemia exhibited symptoms typical of diseases associated with phytoplasmas. In summer 1998 similar symptoms were detected in leek plants in a field used for seed production located in Romagna, North Italy. In both cases the plants were established in the spring of the previous year. Plants showed flower abnormalities: stamen elongation, anther sterility, pistil proliferation, as well as poor, if any, seed production. Phytoplasma-like structures were detected by scanning and transmission electron microscopy in phloem sieve elements in the Czech diseased plants, but not in healthy ones. Nested-PCR amplifications of extracted DNA with phytoplasma-specific oligonucleotide primer pairs confirmed the presence of phytoplasmas in these plants at low concentrations. Restriction fragment length polymorphism analyses of amplified ribosomal sequences allowed the identification of detected phytoplasmas: all the samples from the Czech Republic contained aster yellows related phytoplasmas (16SrI-B) while in the Italian samples aster yellows related phytoplasmas (16SrI-B) together with stolbur related phytoplasmas (16SrXII-A) were identified. This is the first report of detection and identification of a phytoplasma disease of leek in the Czech Republic and Italy.     

家鸡蛋壳组织结构的扫描电镜观察

用扫描电子显微技术对家鸡蛋壳结构进行研究的结果，进一步直观形象地丰富了我们在这个领域的知识。