Detection and characterization of phytoplasmas in diseased stone fruits and pear by PCR-RFLP analysis in Turkey

During the late summer-early autumn of 2002, surveys were carried out in Turkey to determine the presence of phytoplasma diseases in fruit trees. Phytoplasmas were detected and characterized by PCR-RFLP analysis and TEM technique in stone fruit and pear trees in the eastern Mediterranean region of the country. Six out of 24 samples, including almond, apricot, peach, pear and plum, gave positive results in PCR assays. RFLP analysis usingSspI andBsaAI enzymes of PCR products obtained with primer pair f01/r01 enabled identification of the phytoplasmas involved in the diseases. Stone fruit trees, including a local apricot variety (‘Sakıt’) and a pear sample, were found to be infected with European stone fruit yellows (ESFY, 16SrX-B) and pear decline (PD, 16SrX-C) phytoplasmas, respectively. This is the first report in Turkey of PD phytoplasma infecting pear and of ESFY phytoplasma infecting almond, apricot, myrobalan plum and peach; ESFY phytoplasma infecting Japanese plum was previously reported. http://www.phytoparasitica.org posting July 21, 2005.     

Hydrogen peroxide localization and antioxidant status in the recovery of apricot plants from European Stone Fruit Yellows

Hydrogen peroxide (H₂O₂) localization and roles of peroxidases, malondialdehyde and reduced glutathione were compared in leaves of apricot (Prunus armeniaca) plants asymptomatic, European Stone Fruits Yellows (ESFY)-symptomatic and recovered. Nested PCR analysis revealed that Candidatus Phytoplasma prunorum, is present in asymptomatic, symptomatic and recovered apricot trees, confirming previous observations on this species, in which recovery does not seem to be related to the disappearance of phytoplasma from the plant.H₂O₂was detected cytochemically by its reaction with cerium chloride, which produces electron-dense deposits of cerium perhydroxides. H₂O₂was present in the plasmalemma of the phloem cells of recovered apricot plant leaves, but not in the asymptomatic or symptomatic material. Furthermore, by labelling apricot leaf tissues with diaminobenzidine DAB, no differences were found in the localization of peroxidases.Protein content in asymptomatic, symptomatic and recovered leaves was not significantly different from one another. In contrast, guaiacol peroxidase activity had the following trend: symptomatic > recovered > asymptomatic, whereas reduced glutathione content followed the opposite trend: asymptomatic > recovered > symptomatic. Moreover, no differences were observed in malondialdehyde concentrations between asymptomatic, symptomatic and recovered leaves. The overall results suggest that H₂O₂ and related metabolites and enzymes appear to be involved in lessening both pathogen virulence and disease symptom expression in ESFY-infected apricot plants.     

Impaired control of IRES-mediated translation in X-linked dyskeratosis congenita

The DKC1 gene encodes a pseudouridine synthase that modifies ribosomal RNA (rRNA). DKC1 is mutated in people with X-linked dyskeratosis congenita (X-DC), a disease characterized by bone marrow failure, skin abnormalities, and increased susceptibility to cancer. How alterations in ribosome modification might lead to cancer and other features of the disease remains unknown. Using an unbiased proteomics strategy, we discovered a specific defect in IRES (internal ribosome entry site)-dependent translation in Dkc1(m) mice and in cells from X-DC patients. This defect results in impaired translation of messenger RNAs containing IRES elements, including those encoding the tumor suppressor p27(Kip1) and the antiapoptotic factors Bcl-xL and XIAP (X-linked Inhibitor of Apoptosis Protein). Moreover, Dkc1(m) ribosomes were unable to direct translation from IRES elements present in viral messenger RNAs. These findings reveal a potential mechanism by which defective ribosome activity leads to disease and cancer.     

Iron oxide‐humic acid coprecipitates as iron source for cucumber plants

In soil, iron (Fe) solubility depends on complex interactions between Fe minerals and organic matter, but very little is known about plant availability of Fe present in Fe oxides associated with humic substances. For this purpose, this study investigates the effect of Fe mineral crystallinity in the presence of humic acids (HA) on Fe availability to plants. Four Fe–HA mineral coprecipitates were prepared, either in the presence or absence of oxygen, i.e., two goethite (G)‐HA samples containing large amounts of Fe as nanocrystalline goethite and ferrihydrite mixed phases, and two magnetite (M)‐HA samples containing crystalline magnetite. Bioavailability studies were conducted in hydroponic systems on cucumber plants (Cucumis sativus L.) grown under Fe deficient conditions and supplied with the Fe–HA coprecipitates containing goethite or magnetite. Results showed that plants grown in the presence of Fe–HA coprecipitates exhibited a complete recovery from Fe deficiency, albeit less efficiently than plants resupplied with Fe‐chelate fertilizer used as control (Fe‐diethylene triamine penta acetic acid, Fe‐DTPA). However, the supply with either G‐ or M–HA coprecipitates produced different effects on plants: G–HA‐treated plants showed a higher Fe content in leaves, while M–HA‐treated plants displayed a higher leaf biomass and SPAD (Soil–Plant Analysis Development) index recovery, as compared to Fe‐DTPA. The distribution of macronutrients in the leaves, as imaged by micro X‐ray fluorescence (µXRF) spectroscopy, was different in G–HA and M–HA‐treated plants. In particular, plants supplied with the poorly crystalline G–HA coprecipitate with a lower Fe/HA ratio showed features more similar to those of fully recovered plants (supplied with Fe‐DTPA). These results highlight the importance of mineral crystallinity of Fe–HA coprecipitates on Fe bioavailability and Fe uptake in hydroponic experiments. In addition, the present data demonstrate that cucumber plants can efficiently mobilize Fe, even from goethite and ferrihydrite mixed phases and magnetite, which are usually considered unavailable for plant nutrition.     

Iron chlorosis paradox in grapevine

Vitis vinifera L. cv. Pinot blanc was grown in both pot and commercial vineyard under lime‐stress conditions. Dark green to very chlorotic young leaves were sampled in order to analyze chlorophyll and iron (Fe) concentration and content, area, fresh and dry weight, specific weight per area. Iron content per individual leaf was a good parameter to separate dark green leaves from chlorotic ones in both trials, while Fe concentration on DW basis was not. Chlorosis occurrence always decreased leaf dry weight and specific weight per area.     

Localization, morphology, and immunohistochemistry of spinal cord and dorsal root ganglion neurons that innervate the gastrocnemius and superficial digital flexor muscles in cattle

OBJECTIVE: To determine the location, morphology, and neurochemical code of spinal cord and dorsal root ganglion neurons that innervate the gastrocnemius muscle (GM) and superficial digital flexor muscle (FDSM) in cattle. ANIMALS: 5 healthy Friesian male calves. Procedure: 2 different types of neuronalretrograde fluorescent tracers (fast blue and diamidino yellow) were injected into the GM and FDSM, respectively. The neurochemical code (substance P, calcitonin gene-related peptide, galanin, and neuronal nuclear protein) of labeled neurons was investigated by immunohistochemistry. RESULTS: Neurons innervating the GM and FDSM were located along the L6-S2 spinal cord segments and ganglionic levels. A cranial-to-caudal topographic distribution for each muscle was found, indicating that the motor nuclei of the 2 muscles are organized by a somatotopic pattern. The GM and FDSM motoneurons were immunoreactive only for calcitonin gene-related peptide, whereas the afferent neurons were immunoreactive for all of the neurochemical markers considered. CONCLUSION AND CLINICAL RELEVANCE: In our study, location and the extent of neurons that supply the GM and FDSM of cattle were characterized completely. Because the GM and FDSM are involved in spastic paresis of calves and it is thought that spastic paresis results from an excessive activity of the neuromuscular spindle reflex arc, findings in our study may be useful for further electrophysiologic and clinical studies. Knowledge of the neurochemical code of neurons that supply the GM and FDSM in healthy calves could be used to compare chemical alterations in the same neuronal population of affected calves.     

High tolerance of European plum varieties to plum leptonecrosis

A 13 year comparative study was carried out on the behaviour of four European and two Japanese plum varieties grown in adjacent rows in an area of northern Italy where plum leptonecrosis is epidemic. Within seven years, 100% of the Japanese plum trees became symptomatically infected. Nine years after planting, five trees of each of the European cvs, which were asymptomatic, were top-grafted with healthy buds of the cv Ozark Premier, which is an indicator for plum leptonecrosis. Based on the results of PCR analysis, DAPI staining and on the reaction of the top-grafted Ozark Premier indicators, 50% of the European plum trees, despite their healthy appearance, were shown to be infected with plum leptonecrosis. The detectable presence and graft transmissibility of the plum leptonecrosis phytoplasma in the asymptomatic European plum trees means that the European plum trees are not resistant to the infection but that they are tolerant. The active presence of a still unknown vector/s in the investigated area is stressed as well as the important role of Prunus domestica L. played in the conservation and spread of plum leptonecrosis.     

Dyskeratosis congenita and cancer in mice deficient in ribosomal RNA modification

Mutations in DKC1 cause dyskeratosis congenita (DC), a disease characterized by premature aging and increased tumor susceptibility. The DKC1 protein binds to the box H + ACA small nucleolar RNAs and the RNA component of telomerase. Here we show that hypomorphic Dkc1 mutant (Dkc1m) mice recapitulate in the first and second generations (G1 and G2) the clinical features of DC. Dkc1m cells from G1 and G2 mice were impaired in ribosomal RNA pseudouridylation before the onset of disease. Reductions of telomere length in Dkc1m mice became evident only in later generations. These results suggest that deregulated ribosome function is important in the initiation of DC, whereas telomere shortening may modify and/or exacerbate DC.     

Different responses to root infection with endophytic microorganisms of vitis vinifera L. cv. pinot blanc grown on calcareous soil

One‐year‐old rooted cuttings of Vitis vinifera L. cv. “Pinot blanc”; grafted on “Kober 5BB”; hybrid rootstock were grown in pots containing a calcareous soil. Before potting, the roots were treated by a suspension of the siderophore producing bacteria Pseudomonas fluorescens, other Pseudomonas spp., and some unidentified microorganisms. The tested bacteria were endophytic microorganisms from maize and they were chosen according to their ability to produce fluorescent pigments. The shoot growth was checked every ten days, and leaf blades, sampled about 80 days after bud burst, were analyzed for chlorophyll and mineral element concentration. Chlorosis rating was checked by visual screening in order to control the effect of the treatments on the chlorosis. At the end of the growing cycle dry matter and iron (Fe) partitioning were analyzed. The most significant findings of the trial were: a) the tested microorganisms did not improve significantly lime‐tolerance of the tested plants; b) the microorganisms were able to modify phosphorus (P), potassium (K), and Fe mineral nutrition of the vines.     

Differentially-regulated defence genes in Malus domestica during phytoplasma infection and recovery

To improve knowledge about plant/phytoplasma interactions and, in particular, about the ‘recovery’ phenomenon in previously-infected plants, we investigated and compared expression levels of several defence-related genes (four pathogenesis-related proteins and three jasmonate-pathway marker enzymes) in apple plants showing different states of health: vigorous (healthy), phytoplasma-infected, and recovered. Real Time-PCR analyses demonstrated that genes are differentially expressed in apple leaf tissue according to the plants’ state of health. Malus domestica Pathogenesis-Related protein (MdPR) 1, MdPR 2 and MdPR 5 were significantly induced in leaves of diseased and symptomatic plants compared to leaves of those plants that were healthy or recovered. On the other hand, levels of all the jasmonate (JA)-pathway marker genes that we selected for this study, were up-regulated in the leaves of recovered plants compared to the diseased ones. In conclusion, our study demonstrated that two different sets of defence genes are involved in the interactions between apple plants and ‘Candidatus Phytoplasma mali’ (‘Ca. P. mali’) and that these genes are differentially expressed during phytoplasma infection or recovery.