Is there a future for wild grapevine (Vitis vinifera subsp. silvestris) in the Rhine Valley?

The wild grapevine, Vitis vinifera L. subsp. silvestris (Gmelin) Hegi, is considered to be an endangered taxon in Europe, mainly as a consequence of the introduction of pathogens from North America and of the destruction of its habitat. In the Rhine Valley, nearly all populations disappeared due to river management, the intensi.cation of forestry, and the introduction of phylloxera. After a growing awareness of the need to preserve endangered forest ecosystems, attempts to reintroduce wild grapevine in the Rhine Valley were performed, particularly in the French nature reserves Erstein and Offendorf since 1992. However, regular surveys of the plants indicate the rapid decline of the populations. In 2002, we proposed to summarise the knowledge accumulated after 10 years of experiments. Results indicate that from the initial 91 individuals planted in 1992, only 14 survived in 2002 (2 in Erstein, 12 in Offendorf). The failure of the experiment may be explained by several factors: unsuitable sites (too shady, absence of support for the young plants), absence of monitoring, vandalism or predation. According to these results and recent knowledge of the ecology of the plant and of vines in general, new transplantation experiments are proposed in which the plants will be monitored during their establishment in the forests. The success of this second transplant (50 plants per reserve) will be enhanced by restoration projects of the Rhine River dynamics, with partial re-.ooding. Floods should help to avoid, or at least to reduce, pest and disease expansion on future adult plants.     

Effect of phyllotactic position and cultural treatments toward successful direct shoot organogenesis in dwarf ‘Pixie’ grapevine (Vitis vinifera L.)

In Vitis spp. where somatic embryogenesis-based regeneration predominates, an efficient, reproducible and robust method of direct shoot organogenesis from leaf explant material has been established in the dwarf wine grape ‘Pixie’ (Vitis vinifera). This regeneration system was achieved by testing the response of leaf material in two stages of development, and pre-conditioning the explant material in dark conditions and/or in liquid media prior to excising from the plant and placing it on solidified media. The pre-excision treatments included (1) a dark period of 24 h, with no regeneration medium; (2) soaking in regeneration medium followed by a dark period of 24 h; (3) a dark period of 24 h followed by soaking in liquid VRM (Vitis Regeneration Medium); (4) vacuum infiltration in liquid VRM followed by a dark period of 24 h; and (5) a control of no pre-conditioning treatment. Excised leaves from pre-treated intact plants in vitro significantly increased the frequency of shoot organogenesis. The most responsive explant material consisted of young semi-translucent apical leaves varying in size from 3 to 8 mm in length. The most successful combinations of factors contributing to shoot organogenesis involved the solely dark-exposed apical leaves or the soaking in VRM followed by a dark period. These results are expected to facilitate Vitis-related research in genetics, functional genomics, physiology, and other fields.     

Effect of the rootstock on the occurrence of lime-induced chlorosis of potted Vitis vinifera L. cv. ‘Pinot blanc’

The chlorosis susceptible Vitis vinifera L. cv. Pinot blanc was grafted on two hybrid rootstocks with different iron efficiency, as follows: V. Berlandieri × V. rupestris 140 Ru (iron-efficient) and V. riparia × V. rupestris 101-14 (iron-inefficient). The grafted vines were grown in pots of a calcareous and a non-calcareous soil. The shoot growth was periodically checked and leaves, selected at two different times (at the middle of the annual growing period), were assayed for total chlorophyll, ferrous iron, ash alkalinity, percentage of dry matter and chlorosis score. At the end of the growing cycle the roots were oven-dried and weighed. The most significant findings of the trial were: (a) the soil strongly affected the shoot growth, with canes about twice as long in the non-calcareous soil; (b) the iron-efficient rootstock (140 Ru) did not induce chlorosis when growing on the calcareous soil, while the opposite occurred with the iron-inefficient rootstock (101=14); and (c) a high ash alkalinity occurred in light chlorotic leaves compared to green ones, under the same iron concentration.     

Discrimination between cultivars of Vitis vinifera based on molecular variability concerning 5″ untranslated regions of the StSy-CHS genes

 The degree of polymorphism present in 5′ untranslated regions of stilbene synthase (StSy)-like loci was assessed. A ligation-mediated polymerase chain reaction (LM-PCR) cloning strategy was adopted to isolate sequences located immediately upstream of StSy coding regions. Among several clones, 13 randomly chosen fragments were analyzed at the sequence level. Four of the analyzed fragments appeared of particular interest. Two carried sequences reminiscent of micro-satellites, while the remaining fragments contained direct repeats. Oligonucleotides constructed against the specific DNA sequence of these clones disclosed a complex banding pattern when used in polymerase chain reaction (PCR)-analysis of 22 ancient varieties of grapevine. A total of 40 polymorphic bands could be identified and used to calculate coefficients of genetic similarity (GS) between varieties. GS values were used in cluster analysis to differentiate the 22 varieties. The data obtained are in good agreement with available information concerning the relationships between the varieties considered. This suggests the use of the method we have developed in fingerprinting studies of Vitis vinifera germ plasma. Received: 11 April 1996 / Accepted: 14 March 1997     

Response of the Vitis vinifera L. cv. 'Nebbiolo' proteome to Flavescence dorée phytoplasma infection

Flavescence dorée is a serious phytoplasma disease affecting grapevine in several European countries. We studied the interaction of Flavescence dorée phytoplasma with its natural plant host by monitoring the effects of infection on the protein expression profile. Among the 576 analyzed spots, 33 proteins were differentially regulated in infected grapevines. Grouping into MIPS functional categories showed proteins involved in metabolism (21%), energy processes (9%), protein synthesis (3%), protein fate (18%), cellular transport and transport routes (6%), cell defense and virulence (42%). Among the differentially regulated proteins, we selected six targets (thaumatin I, thaumatin II, osmotin-like protein, plant basic secretory protein, AAA(+) Rubisco activase and proteasome α5 subunit) and we analyzed their expression by quantitative RT-PCR on samples collected in 2008 and 2009 in several vineyards in Piedmont region, Italy. There was a positive correlation between mRNA and protein expression for most of the genes in both the years. We discuss the involvement of these proteins in the specific response to phytoplasma infection. To our knowledge, this work is the first to investigate the response of the grapevine proteome to Flavescence dorée phytoplasma infection, and provides reference protein profiles for future comparative proteomic and genomic studies.     

Dynamic distribution of TTK in HeLa cells： insights from an ultrastructural study

Entry into mitosis is driven by signaling cascades of mitotic kinases. Our recent studies show that TTK, a kinetochore-associated protein kinase, interacts with CENP-E, a mitotic kinesin located to corona fiber of kinetochore. Using immunoelectron microscopy, here we show that TTK is present at the nuclear pore adjacent complex of interphase HeLa cells. Upon nuclear envelope fragmentation, TTK targets to the outermost region of the developing kinetochores of monoorient chromosome as well as to spindle poles. After stable attachment, throughout chromosome congression, TTK is a constituent of the corona fibers, extending up to 90 nm away from the kinetochore outer plate. Upon metaphase alignment, TTK departs from the kinetochore and migrates toward the centrosomes. Taken together, this evidence strongly supports a model in which TTK functions in spindle checkpoint signaling cascades at both kinetochore and centrosome.     

Effects of Shading on the Synthesis of Volatile Organic Compounds in ‘Marselan’ Grape Berries (Vitis vinifera L.)

Journal of Plant Growth Regulation - Light is a vital environmental factor that can affect the synthesis of volatile organic compounds (VOCs) in grape berries. However, the mechanism through which...     

Eriophyoid mite damage in Vitis vinifera (grapevine) in Australia: Calepitrimerus vitis and Colomerus vitis (Acari: Eriophyidae) as the common cause of the widespread ‘Restricted Spring Growth’ syndrome

Leaf and shoot distortions and retarded shoot growth in Vitis vinifera L. prevalent in Australian vineyards in early spring, were investigated in replicated field experiments over 3 yrs. Leaf distortion and retarded shoot growth were identified as damage due to feeding of extremely high populations of over-wintered deutogynes of Calepitrimerus vitis (Nalepa) (grape rust mite). This damage was hitherto known in Australia as Restricted Spring Growth (RSG), a syndrome comprising several growth abnormality symptoms, none with a clearly identified cause or a successful treatment. A successful treatment against C. vitis was used to selectively eliminate RSG, while C. vitis numbers were recorded using a validated trapping technique; intercepting deutogynes migrating from winter shelters in the wooden vine structure, to emerging green tissues. Severe leaf distortion was associated with >400 C. vitis deutogynes per spur, while >1000 per spur had the added effect of severely retarding shoot growth. A 43.0–47.2% shoot length reduction was recorded for Cabernet Sauvignon, 27.1–32.8% for Sauvignon Blanc, when 4–6 leaves were separated. Symptoms were most prominent up to 8–9 separated leaves, however 24.7–30.4% shoot length reduction was still evident at flowering, and 12.8% circa fruit set. C. vitis effect on vine fruitfulness, and yield parameters at fruit set, were also studied. Once successfully treated to prevent C. vitis damage, poor bud burst remained evident in some vineyards. Surveys of unburst buds from such vineyards revealed presence of Colomerus vitis (Pagenstecher) (grape bud mite). When Col. vitis numbers in unburst buds reached 100–500 per bud, apical meristems of primary, and commonly also secondary buds were dead, preventing bud burst. The remaining living scale tissue was distinctly scarred. Bud and associated shoot damage were documented. Retarded shoot growth and leaf distortion, previously attributed to RSG, are misdiagnosed C. vitis spring feeding damage. Clustered high infestations of Col. vitis can cause bud-axis necrosis, bud burst failure, shoots with short basal internodes, and short, thin, zigzagged shoots with absent fruit clusters; all previously considered RSG.This revised version was published online in May 2005 with a corrected cover date.     

A novel Bacteroidetes symbiont is localized in Scaphoideus titanus, the insect vector of Flavescence dorée in Vitis vinifera

Flavescence dorée (FD) is a grapevine disease that afflicts several wine production areas in Europe, from Portugal to Serbia. FD is caused by a bacterium, "Candidatus Phytoplasma vitis," which is spread throughout the vineyards by a leafhopper, Scaphoideus titanus (Cicadellidae). After collection of S. titanus specimens from FD-contaminated vineyards in three different areas in the Piedmont region of Italy, we performed a survey to characterize the bacterial microflora associated with this insect. Using length heterogeneity PCR with universal primers for bacteria we identified a major peak associated with almost all of the individuals examined (both males and females). Characterization by denaturing gradient gel electrophoresis confirmed the presence of a major band that, after sequencing, showed a 97 to 99% identity with Bacteroidetes symbionts of the "Candidatus Cardinium hertigii" group. In addition, electron microscopy of tissues of S. titanus fed for 3 months on phytoplasma-infected grapevine plants showed bacterial cells with the typical morphology of "Ca. Cardinium hertigii." This endosymbiont, tentatively designated ST1-C, was found in the cytoplasm of previtellogenic and vitellogenic ovarian cells, in the follicle cells, and in the fat body and salivary glands. In addition, cell morphologies resembling those of "Ca. Phytoplasma vitis" were detected in the midgut, and specific PCR assays indicated the presence of the phytoplasma in the gut, fat body and salivary glands. These results indicate that ST1-C and "Ca. Phytoplasma vitis" have a complex life cycle in the body of S. titanus and are colocalized in different organs and tissues.     

Dynamic properties of pH-dependent structural organization of the amyloidogenic ��-protein (1�C40)

The structural organization of the amyloidogenic β-protein containing 40 amino acid residues (Aβ40) was studied by the high temperature molecular dynamics simulations in the acidic (pH ∼ 3) and basic (pH ∼ 8) pH regions. The obtained data suggest that the central Ala21-Gly29 segment of Aβ40 can adopt folded and partially unfolded structures. At the basic pH, this segment forms folded structures stabilized by electrostatic interactions and hydrogen bonds. At the acidic pH, it forms partially unfolded structures. Two other segments flanking to the central segment exhibit the propensity to adopt unstable interconverting α-helical, 3₁₀-helical and turn-like structures. One of these segments is comprised of the Ala30-Val36 residues at both of the considered pHs. The second segment is comprised of the Glu11-Phe20 at the basic pH and of the Glu11-Val24 residues at the acidic pHs. The revealed pH-dependent structuration of the Aβ40 allowed us to suggest a possible scenario for initial Aβ aggregation. According to this scenario, the occurrence of the partially unfolded states of the Ala21-Gly29 segment plays main role in the Aβ oligomerization process.Key words: amyloid-β protein, Alzheimer disease, oligomerization, fibril, electrostatic interactions, molecular dynamics simulations     

DIGE analysis of proteome changes accompanying large resveratrol production by grapevine (Vitis vinifera cv. Gamay) cell cultures in response to methyl-β-cyclodextrin and methyl jasmonate elicitors

We had previously shown that Vitis vinifera cv. Gamay cell suspension accumulates extracellularly large amounts of the phytoalexin trans-resveratrol (tR) in response to elicitation with methylated cyclodextrins (MBCD), which can be triplicated when the elicitor is combined with methyl jasmonate (MeJA). In parallel, new pathogenesis-related proteins accumulated in the apoplast-like extracellular space. The aim of this study was to investigate changes in the grapevine cell proteome potentially related to tR accumulation in response to the above elicitors. The DIGE technique was used to detect statistically significant changes in the cell's proteome. A total number of 1031 unique spots were detected, 67 of which were de-regulated upon elicitation. Sixty-four spots were successfully identified by nLC-MS/MS database search analysis. The tR biosynthetic pathway enzymes were up-regulated by MBCD alone or combined with MeJA, but not by treatment with MeJA alone, in agreement with tR accumulation pattern. Seven spots contained stilbene synthase encoded by four different isogenes. Likewise, four glutathione-S-transferases, potentially involved in tR trafficking within the cell and across membranes, were up-regulated in the same fashion as stilbene synthases. The relation of other de-regulated proteins with other effects caused by elicitors on grapevine cells, namely defense response and cell growth inhibition, is discussed.     

Dynamic complexity of visuo-motor coordination: an extension of Bernstein’s conception of the degrees-of-freedom problem

 Extending Bernstein’s spatial conception of the degrees-of-freedom problem in the human motor system, we introduce a method developed from the theory of non-linear dynamics that allows one to quantify the spatio-temporal, i.e. dynamic, complexity of visuo-motor coordination. The correlation dimension D is used to measure the effective number of dynamic degrees of freedom in the coordination that a subject uses when performing a visuo-motor tracking task. The validity of the estimator employed is demonstrated. Visuo-motor coordination had a low-dimensional (mean D±SD=6.07 ±0.82) dynamic structure, which was consistent with deterministic chaos rather than with pure stochastic noise. D correlated with tracking performance, P. Both D and P were closely related to the degree of visuo-motor compatibility that the task presented to the subject. However, for short periods of training P increased, but D did not. As these seemingly contradictory results suggest, our dynamic conception of the degrees-of-freedom problem may reveal far more intricate visuo-motor interactions than Bernstein could identify on the basis of his spatial analyses of bodily movement patterns and by the methods of evaluation that were available to him at the time. Received: 19 April 1995/Accepted in revised form: 17 June 1996     

Do exogenous polyamines have an impact on the response of a salt-sensitive rice cultivar to NaCl?

In order to analyze the putative impact of polyamines (PAs) on the plant response to salt, seedlings from the salt-sensitive rice cultivar I Kong Pao (IKP) were exposed for 5, 12 and 19 days to 0, 50 or 100 mM NaCl in the absence, or in the presence of exogenous PAs (putrescine (Put), spermidine (Spd) or spermine (Spm) 1mM) or inhibitors of PA synthesis (methylglyoxalbis-guanyl hydrazone (MGBG) 1mM, cyclohexylammonium (CHA) 5mM and D-arginine (D-Arg) 5mM). The addition of PAs in nutritive solution reduced plant growth in the absence of NaCl and did not afford protection in the presence of salt. PA-treated plants exhibited a higher K+/Na+ ratio in the shoots, suggesting an improved discrimination among monovalent cations at the root level, especially at the sites of xylem loading. The diamine Put induced a decrease in the shoot water content in the presence of NaCl, while Spd and Spm had no effects on the plant water status. In contrast to Spd, Spm was efficiently translocated to the shoots. Both PAs (Spd and Spm) induced a decrease in cell membrane stability as suggested by a strong increase in malondialdehyde content of PA-treated plants exposed to NaCl. These results are discussed in relation to the putative functions of PAs in stressed plant metabolism.