New treatments to disinfect vine cuttings with Phaeomoniella chlamydospora and the reason for the control

Petri disease is a problem for vineyard caused mainly by the fungus Phaeomoniella chlamydospora. Contaminated seedlings are source of inoculum for the disease. Treatment to disinfect vine rootstock cuttings for seedling production is hot water treatment (HWT) by 50 °C for 30 min, but the efficiency is contested. To improve its efficacy, the study aimed to assess the combination of the following methods and the reason for the control: i) exposition of the fungus to five different temperatures in HWT bath for 30 min; ii and iii) exposition of the fungus and also plants infected with P. chlamydospora to different disinfection treatments (biofumigation = soil + cabbage at 40 °C; temperatures of 40 and 23 °C, all in microcosm), in different periods (7, 14 and 21 days), with and without additional HWT (51 °C for 30 min). The results showed that HWT with high temperatures (55­70 °C) for 30 min inactivated the fungus. Biofumigation technique at 40 °C and the temperature solely of 40 °C applied for up to 21 days and combined with HWT (51 °C for 30 min) inhibited mycelial growth and inactivated the fungus in vine plant tissues without compromising the rooting.

Dual effects of entomopathogenic fungi on control of the pest Lobesia botrana and the pathogenic fungus Eutypella microtheca on grapevine

BACKGROUND: Entomopathogenic fungi (EPF) are the natural enemies of insect pests. Nevertheless, research on the use of EPF for simultaneous prevention of pest and disease agents on the same crop is limited. In this study, we explored the potential dual effects of three strains of the EPF Metarhizium anisopliae on the control of detrimental agents of Vitis vinifera L., including different developmental stages (larvae, pupae, and adult) of the insect pest Lobesia botrana and the phytopathogenic fungus Eutypella microtheca. METHODS: Laboratory pathogenicity trials were performed to examine the effects of the three M. anisopliae strains on the mortality rate of L. botrana. In addition, field trials were conducted to assess the biocontrol potential of one selected M. anisopliae strain on the larval stage of L. botrana. Moreover, inhibitory effects of the three EPF strains on E. microtheca growth were examined in vitro. RESULTS: All the M. anisopliae strains were highly effective, killing all stages of L. botrana as well as inhibiting the growth of E. microtheca. The in vitro mortality of larvae treated with the strains was over 75%, whereas that of treated pupae and adults was over 85%. The three EPF strains showed similar efficacy against larvae and adult stages; never-theless, pupal mortality was observed to be strain dependent. Mortality of L. botrana larvae ranged from 64 to 91% at field conditions. Inhibition of E. microtheca growth reached 50% in comparison to the control. CONCLUSIONS: Our study showed that M. anisopliae strains were highly effective in ensuring control of two different detrimental agents of V. vinifera L., providing new evidence to support the dual effects of entomopathogenic fungi.

Growth supression of plant pathogenic fungi using bryophite extracts / Supressão de crescimento de fungos patogênicos de plantas usando extratos de briófitas

Chemicals are often used in attempts to control diseases caused by plant pathogenic fungi during food production. However, chemicals can have adverse effects not just on food, but they also remain active for a long time within ecosystems, and thus are not environmentally friendly. Therefore,development of bio-treatment and avoiding use of chemicals are urgently needed. With the aim of studying and developing new environmentally-friendly treatments, we tested extracts from selected bryophyte species(Porella platyphylla, Cinclidotus fontinaloides and Anomodon viticulosus) on five plant pathogenic fungi under controlled conditions. The fungi (Botryosphaeria dothidea, Phomopsis viticola, Calosphaeria sp., Colletotrichum acutatum and Monilinia laxa) were selected based on common diseases they cause on fruits and grapevine. They were isolated in cultures and treated with bryophyte extracts. Bryophyte extracts were shown to be effective in suppression of certain plant pathogenic fungi growth and to have a huge potential in development of novel biotechnological treatments and biofungicides. The best results were achieved in inhibition of B. dothidea, P. viticola and Calosphaeria sp.

Fungos fitopatogênicos são controlados com produtos químicos para combater doenças causadas por eles durante a produção de alimentos. Tais produtos são ruins não apenas para a alimentação, mas também podem permanecer por muito tempo nos ecossistemas, portanto, não são ecologicamente corretos. Desta forma, os biotratamentos e a prevenção de usos químicos são urgentemente necessários no futuro próximo. Com o objetivo de estudar e desenvolver nova alternative decontrole de doenças de plantas, testamos os extratos de espécies de briófitas selecionadas (Porella platyphylla, Cinclidotus fontinaloides e Anomodon viticulosus) em cinco fungos fitopatogênicos (Botryosphaeria dothidea, Phomopsis viticola, Calosphaeria sp., Colletotrichum acutatum e Monilinia laxa) em condição controlada. Estes fungos, selecionados com base nas doenças que causam em frutos e videiras, foram isolados em culturas puras e tratados com os respectivos extratos de briófitas. Os extratos de briófitas demonstraram ser eficazes na supressão de certos fungos fitopatogênicos e têm um enorme potencial no desenvolvimento de novos biofungicidas e tratamentos biotecnológicos. Os mais interessantes resultados foram obtidos na inibição de B. dothidea, P. viticola eCalosphaeria sp.

Yield and physicochemical characteristics of 'BRS Magna' and 'Isabel Precoce' grapes influenced by pruning in the SãoFrancisco river valley / Produtividade e características físico-químicas das uvas 'BRS Magna' e 'Isabel precoce' influenciadas pela poda no Vale do São Francisco

ABSTRACT: The objective of this study was to evaluate the productive performance and physicochemical characteristics of 'BRS Magna' and 'Isabel Precoce' grapes influenced by two types of pruning. The experiment was carried out during two growing seasons in a commercial vineyard of Petrolina, PE, in the period 2013-2014. Treatments were represented by two cultivars, 'Isabel Precoce' and 'BRS Magna', and two types of pruning, with selection during plant formation (P1) and without selection (P2) of lateral branches, using a randomized experimental block design, in subdivided plots and four replicates. After pruning P2, the amount of buds kept on vines was greater than after pruning P1, favoring an increase in yield. However, the sizes of bunches and berries, content of soluble solids, and titratable acidity were not influenced by the pruning type. In general, the sizes of bunches and berries in 'BRS Magna' were larger than in the 'Isabel Precoce' cultivar. Conversely, content of soluble solids and titratable acidity varied among cultivars as a function of production cycle and type of pruning; however, within the standards described for both cultivars and accepted by the Brazilian legislation.

RESUMO: O presente trabalho teve como objetivo estudar o desempenho produtivo e características físico-químicas das uvas 'BRS Magna' e 'Isabel Precoce' submetidas a dois tipos de poda. Um experimento foi realizado durante dois ciclos de produção, nos anos de 2013 e 2014, em um vinhedo comercial em Petrolina, PE. Os tratamentos foram representados por duas cultivares copa, 'Isabel Precoce' e 'BRS Magna', e dois tipos de poda, com seleção de ramos laterais na formação da planta (P1) e sem seleção de ramos laterais (P2), utilizando delineamento experimental em blocos ao acaso em parcelas subdivididas e quatro repetições. A quantidade de gemas mantidas na videira após a poda é mais elevada na poda P2, favorecendo o aumento da produtividade, mas o tamanho dos cachos e das bagas e o teor de sólidos solúveis e acidez titulável não foram influenciados pelo tipo de poda. O tamanho dos cachos e das bagas da cultivar 'BRS Magna' é, de modo geral, maior que os de 'Isabel Precoce'. Por outro lado, o teor de sólidos solúveis e acidez titulável variaram entre as cultivares em função do ciclo de produção e do tipo de poda, estando dentro dos padrões descritos para ambas cultivares e aceitos pela legislação brasileira.

In vitro propagation of Vitis vinifera L. cv. 'Monastrell'

Background: A protocol for the micropropagation of the grape (Vitis vinifera L.) cultivar 'Monastrell' was developed. Initial plant material was obtained from the sanitary selection of grapevine plants performed by real-time RT-PCR to confirm the absence of Grapevine fanleaf virus, Arabis mosaic virus, Grapevine leafroll-associated virus 1, Grapevine leafroll-associated virus 3, and Grapevine fleck virus. Results: The effects of the salt composition (comparing Lloyd and McCown woody plant medium and Murashige and Skoog medium 1/2 macronutrients) and the growth regulator benzylaminopurine (BAP), at 0 and 8.9 µM, on plant propagation were evaluated using nodes as explants. The most efficient procedure consisted of bud induction in the medium with Lloyd and McCown woody plant salts and 8.9 µM BAP for 30 d along with elongation in cytokinin-free medium for 60 d, which gave 22 nodes/explant (174 plants/initial plant). A second cycle of propagation in a medium without BAP for another 60 d could give approximately 10,000 nodes, which can be obtained after an additional 2 months of culture. All plants acclimatized after the second cycle of multiplication were successfully transferred to soil. Conclusion: We developed an optimal protocol for V. vinifera cv. 'Monastrell' micropropagation, the first described for this cultivar.

Determination of polyphenols and antioxidant activity of Vitis labrusca cv. baile berries.

Grape juice and grape skin extracts are important commercial source of polyphenolic compounds which exert different functional properties such as color potential, antimicrobial, antioxidant activity, and health benefits. In this paper we describe a sensitive and specific assay for determination of bioactive polyphenolic compounds in Campbell Early (Vitis labrusca cv. baile). Five polyphenolic components were separated on an Agilent Zorbax Extend C18 Column (250 mm × 4.6 mm × 5 μm) and detected by a diode array detector. The mobile phase was composed of (a) aqueous phosphoric acid (0.2%, v/v); and (b) acetonitrile using a gradient elution. Analytes were performed at 25°C with a flow rate of 0.8 ml/min and UV detection at 280, 360, and 520 nm. All calibration curves showed good linear regression (r2 ≥ 0.9999) within tested ranges. Overall intra- and inter-day variations were less than 1.90%, and the average recoveries were 95.5-105% for analytes. The antioxidant activity determined by DPPH radical assay, ranged from 86-105 for extracts, and 165-252 for studied standards (µM trolox/100 g dry wt.). The proposed method would be sensitive enough and reliable for quality control in functional food and modernization of Campbell Early (Vitis labrusca cv. baile) as potent antioxidant agent.

First record of qualitative losses caused by Meghimatium pictum in vineyards of Southern Brazil and the effects of two molluscicides for its control / Primeiro Registro de perdas qualitativas causadas por Meghimatium pictum em vinhedos do Sul do Brasil e os efeitos de dois moluscicidas para o seu controle

This research reports the terrestrial slug Meghimatium pictum (Stoliczka, 1873) (Stylommathophora: Philomycidae) as an agricultural pest for the first time in Southern Brazil vineyards. The species was found in densities exceeding 20 slugs.m-2 in Vitis labrusca L. vineyards at six municipalities of the Southern Brazil's viticulture region. It causes damage a loss by leaving residual mucus on grapes and by consuming grapes already perforated by other organisms, such as insects or birds, or mechanically damaged by in situ compression. The effectiveness of iron phosphate and metaldehyde baits on M. pictum was evaluated in laboratory experiments. Iron phosphate bait was more effective in controlling M. pictum (70%) than metaldehyde bait (15%).

O presente trabalho relata pela primeira vez a lesma terrestre Meghimatium pictum (Stoliczka, 1873) (Stylommathophora: Philomycidae) como uma praga agrícola causando danos em vinhedos no Sul do Brasil. Esta espécie foi encontrada em densidades superiores a 20 lesmas.m-2 danificando uvas da espécie Vitis labrusca L. em seis municípios da região vitícola do Sul do Brasil. O impacto econômico causado por M. pictum está associado ao movimento dos espécimes no dossel da videira e nas uvas, provocando a contaminação residual por muco e o consumo de uvas já perfuradas por outros organismos, tais como insetos ou aves, ou mecanicamente danificadas pela compressão in situ. A eficácia das iscas a base de fosfato de ferro e metaldeído sobre M. pictum também foi avaliada em condições de laboratório. A isca a base de fosfato de ferro foi mais eficiente no controle de M. pictum (70%) do que a isca a base de metaldeído (15%).

Influence of different pretreatment methods on stilbenoids in grapevine / 中草药

Objective: To study the influence of different pretreatment methods of grapevine on the content of stibenoids in grapevine. Methods: The concentration of resveratrol and ε-viniferin was determined by HPLC, the fresh grapevine sample after picked was pretreated by different methods such as drying in the shade, drying in a dryer under different temperature, keeping in cold storage, slicing, pulverization, or whole treating, and the dynamic changes of stilbenoids in grapevine was investigated. Results: The contents of resveratrol and ε-viniferin in the fresh sample were very low, but the contents of resveratrol and ε-viniferin in the grapevine after picked would change significantly if treated as drying in the shade with whole branch, especially the content of resveratrol would rise from 0.063 mg/g to 7.175 mg/g with the extension of treating time, and the content of ε-viniferin would rise from 0.838 mg/g to 4.225 mg/g, but at some extent, the content would decline. The contents of resveratrol and ε-viniferin would not rise if the sample was pretreated as drying in the shade after slicing, drying in a dryer or keeping in cold storage. Conclusion: The influence of different pretreatment methods to the contents of resveratrol and ε-viniferin is very large, during drying process in the shade with whole branch, the contents of resveratrol and ε-viniferin change obviously and display the tendency of increasing at the earlier phase of the drying process, and then decrease with the extension of time. The contents of resveratrol and ε-viniferin can reach the level of utilization with the pretreatment of drying process in the shade.

Cisgenesis and Intragenesis: New tools For Improving Crops

Genetically Modified Organisms (GMO) could be the answer for many relevant problems affecting crops. However, improving crops through GMO is also often associated with safety concerns, environmental risks and health issues due to the presence of foreign DNA. These limitations have prompted the development of alternative technologies. Recently, cisgenesis and intragenesis have been developed as new tools aimed to modify crops. While cisgenesis involves genetic modification using a complete copy of natural genes with their regulatory elements that belong exclusively to sexually compatible plants, intragenesis refers to the transference of new combinations of genes and regulatory sequences belonging to that particular species. So far, application of cisgenesis and intragenesis as alternatives to conventional transgenesis are limited to a few species, mainly due to the lack of knowledge of the regulatory sequences required. The grape is one of the most cultivated crops worldwide and is the most economically relevant crop in Chile. Its genomic sequence has been completed, making available new sources of information to improve grape traits by genetic manipulation. This review is focused on the current alternatives to transgenesis in plants, including new approaches to develop marker-free crops, their application to economically relevant crops and future perspectives in the area. Also, the identification of grapevine promoters with a wide range of expression profiles is shown. The expression pattern of these genes was analyzed in different tissues and developmental stages, as well as under several stresses and stimuli, giving a broad range of expression patterns, including genes expressed exclusively during ripening, in response to sugars, senescence and biotic stress, among others. Genes with strong and constitutive expression were also identified. Functional analysis using reporter genes has been conducted in order to confirm the promoter's transcription activity, opening new possibilities for developing cisgenic/intragenic grapevines.

Study of Aging and Hepatoprotective Activity of Vitis vinifera L. Seeds in Albino Rats

Objective: Present study was conducted to investigate in liver of rats from 8-12 weeks old to 20 weeks old, the age dependent changes, carbon tetrachloride mediated changes, and the hepatoprotective effect shown by the seeds of Vitis vinifera L. Method: The hepatoprotective activity was studied by observing the effect of 100 mg/kg dose of ethanolic extract of grape seeds on carbon tetrachloride induced hepatotoxicity in albino rats and results were compared with those of the aged group results. Results: 100 mg/kg b.w. of ethanolic extract of Vitis vinifera seeds produced highly significant decrease in AST, ALT, ALP, bilirubin, albumin levels and significant decrease in the TSP levels compared to the toxic group levels. The levels of AST, ALT, ALP, bilirubin and albumin in aged control rats were found to be significantly higher than the levels in young control animals. MDA levels were slightly higher while GSH levels were lower in aged control rats as compared to young control rats. MDA levels in the toxic group showed highly significant increase compared to the young control levels. Ethanolic extract of seeds of Vitis vinifera significantly lowered the MDA levels. Histopathology results reveal that 100mg/kg/day dose of ethanolic extract of seeds of Vitis vinifera L. cured the hepatic damage to a great extent which was induced by CCl4. Conclusions: Aging leads to the changes in the hepatic structure which are comparable to the changes induced by low doses of a hepatotoxin and the ethanolic extract of seeds of Vitis vinifera L. was effective in bringing about functional improvement of hepatocytes exposed to free radical attack, which was confirmed by biochemical and histological observations.

Hepatoprotective, cardioprotective, and renal-protective effects of organic and conventional grapevine leaf extracts (Vitis labrusca var. Bordo) on Wistar rat tissues

The purpose of this study was to evaluate the beneficial effects of organic and conventional grapevine (Vitis labrusca L.) leaf extracts in reducing hydrogen peroxide-induced stress in the liver, heart and kidney of Wistar rats by measuring lipids and proteins damages (carbonyl assay), as well as the activity of the antioxidant enzymes superoxide dismutase and catalase. The preincubation with 5 mg/mL of organic and conventional grapevine (Vitis labrusca L.) leaf extracts prevented both lipids and proteins oxidative damages in all tissues analyzed. The organic leaf extract was able to restore superoxide dismutase (kidney and liver) and catalase (heart) activities, which were modified by the treatment with H2O2. The conventional extract was able to restore only the catalase activity in liver and heart tissues. The beneficial effects of the V labrusca leaf extract shown in this study could probably be important for formulating dietary supplements, as well as for developing new ingredients with improved antioxidant properties from other plant sources.

O objetivo deste estudo foi avaliar os efeitos benéficos de extratos de folhas de videira (Vitis labrusca L.) orgânicas e convencionais em reduzir o dano gerado pelo peróxido de hidrogênio no fígado, coração e rim de ratos Wistar, pela medida de danos a lipídios e a proteínas (Ensaio Carbonyl), como também a modulação sob a atividade das enzimas antioxi-dantes superoxido dismutase e catalase. A pré-incubação com 5 mg/mL de extratos de folhas de videira (Vitis labrusca L.) orgânicas e convencionais previnem ambos danos oxidativos a lipídios e proteínas em todos os tecidos analisados. O extrato de folha orgânica foi capaz de restabelecer a atividade das enzimas superóxido dismutase (rim e fígado) e catalase (coração), as quais foram modificadas pelo tratamento com peróxido de hidrogênio. O extrato convencional foi capaz de restabelecer apenas a enzima catalase no fígado e no coração. Os efeitos do extrato da folha V. labrusca mostrados neste estudo, provavelmente, poderiam ser importantes para a formulação de suplementos dietéticos, bem como para o desenvolvimento de novos ingredientes com propriedades antioxidantes provenientes de outras plantas.

Produção e qualidade de uvas de mesa 'Sugraone' sobre diferentes porta-enxertos no Submédio do Vale do São Francisco / Production and quality of table grapes 'Sugraone' on different rootstocks in the São Francisco River Valley

O presente trabalho teve como objetivos avaliar o comportamento agronômico da cultivar de uva de mesa sem sementes Sugraone sobre quatro porta-enxertos nas condições do Submédio do Vale do São Francisco. O experimento foi conduzido em um vinhedo comercial no município de Sento Sé-BA durante quatro ciclos de produção (2002 à 2004), em sistema de condução do tipo latada e irrigação por gotejamento. Os tratamentos consistiram de quatro cultivares de porta-enxerto ('Paulsen 1103', 'SO4', 'Harmony' e '420A'), utilizando-se delineamento experimental em blocos ao acaso. Foram obtidas maiores produções por planta e vigor vegetativo sobre o porta-enxerto 'Paulsen 1103'. Também observou-se uma tendência de maior massa de cachos, massa e tamanho de bagas sobre este porta-enxerto, embora tenham ocorrido variações entre os ciclos de produção. O porta-enxerto exerceu influência sobre o teor de sólidos solúveis totais, mas não afetou a acidez total e a relação SST/ATT dos frutos.

The present work aimed to evaluate the agronomic performance of the seedless grape cultivar Sugraone on four rootstocks in the São Francisco Valley. The trial was carried out in a commercial vineyard in Sento Sé, State of Bahia for four seasons (2002 to 2004). The treatments were four cultivars of rootstock ('Paulsen 1103', 'SO4', 'Harmony' and '420A'), using a randomized block design. The highest yield per plant and vegetative vigor were obtained on the rootstock Paulsen 1103. Also there was a tendency for higher mass of bunches, mass and size of berries on the same rootstock, although there were variations between seasons. The rootstock showed positive influence on the content of soluble solids, but did not affect the total acidity and TSS/TA. Young plants of grapevine cv. Sugraone showed good agronomic performance on Paulsen 1103, but it is important to continue the studies for a larger number of years to recommend their use in the São Francisco Valley.

Evaluation of drought tolerance of new grapevine rootstock hybrids.

The drought tolerance is a very important property of grapevine rootstocks. For that reason the breeding and selection of new rootstock varieties is focused also on the evaluation of their drought tolerance. In this experiment, altogether 20 new hybrids and 4 existing rootstock varieties were compared and evaluated. The experimental scheme involved 3 variants of water supply. Evaluated were the following properties: growth intensity of annual shoots, CCI (chlorophyll content index) and visual characteristics of plants. The most resistant were hybrids from the pedigree groups C (Binova x Börner), D /Binova x /(Binova x Teleki 5C/) x Börner/, and F (Teleki 5 C x Börner). The following hybrids were classified as drought-tolerant: 17-1-6 (C); 17- 1-9 (C); 17-6-2 (C); 17-6-9 (C); 17-8-2 (D) and 9-20-1 (F). Based on obtained experimental results and also on correlations existing between individual traits it can be concluded that practically all traits under study may be used when evaluating the resistance of plants to drought. The obtained results indicated that the Börner rootstock (and thus also the species Vitis cinerea) can be used as a suitable genetic resource for the purpose of the breeding grapevine rootstocks for tolerance to drought.

Detecção e caracterização molecular dos genes da proteína capsidial de ilarvírus e ampelovírus que infectam fruteiras temperadas / Detection and molecular characterization of ilarvirus and ampelovirus coat protein genes infecting temperate fruit trees

Dentre os principais patógenos que incidem em fruteiras temperadas, destacam-se o Prune dwarf virus (PDV), o Apple mosaic virus (ApMV) e o Grapevine leafroll-associated virus 1 (GLRaV-1). Neste trabalho foram realizadas a detecção e a caracterização molecular dos genes da proteína capsidial de isolados destas três espécies virais. RNAs totais foram extraídos de amostras de folhas de pessegueiros, macieiras e videiras e, nas reações de RT-PCR, foram utilizados oligonucleotídeos específicos para cada espécie viral. Os cDNAs amplificados foram clonados e sequenciados. Foram verificadas altas identidades entre as sequências de nucleotídeos dos genes da proteína capsidial dos isolados brasileiros de PDV, ApMV e GLRaV-1 e isolados de outros países, independente da origem geográfica e da hospedeira. O peso molecular da proteína capsidial destes vírus foi estimado por meio de Western blot em cerca de 24kDa (PDV), 26kDa (ApMV) e 39kDa (GLRaV-1).

Among the main pathogens infecting temperate fruit trees are Prune dwarf virus, Apple mosaic virus and Grapevine leafroll-associated virus 1. In this work the detection and molecular characterization of the coat protein genes of isolates from these viral species were carried out. Total RNA was extracted from peach, apple and grapevine leaves and RT-PCR reactions were performed using specific primers to each virus. The amplified cDNA fragments were cloned and sequenced. High identities were observed between coat protein nucleotide sequences of Brazilian isolates of PDV, ApMV and GLRaV-1 and isolates from other countries, independently from geographic origin and host. Coat protein molecular weights of these viruses were estimated by Western blot to be ca. 24kDa (PDV), 26kDa (ApMV) and 39kDa (GLRaV-1).

Efeitos da quitosana no desenvolvimento in vitro de videiras cv. merlot e no crescimento micelial do fungo Elsinoe ampelina / Effects of the chitosan on the development of grapevines cv. Merlot and on the mycelial growth of the fungus Elsinoe ampelina

Objetivou-se, neste trabalho, avaliar o efeito da quitosana no desenvolvimento in vitro de plântulas de videira cv. Merlot e sua atividade antifúngica sobre Elsinoe ampelina. No primeiro experimento, explantes da cultivar Merlot foram transferidos para meio de cultura DSD1, acrescido das concentrações 0; 25; 50,100; 150 e 200 mg L-1 de quitosana. Após 90 dias de cultivo in vitro, as plântulas foram avaliadas quanto ao número de raízes e de folhas, porcentagem de enraizamento e brotação, comprimento de raízes e de parte aérea, massa fresca da planta. No segundo experimento, incorporou-se às concentrações 0, 60, 120, 180, 240 e 300 mg L-1 de quitosana ao meio BDA, onde inoculou-se o fungo. Posteriormente, avaliou-se o crescimento micelial aos 6 e 9 dias de incubação a 25º C no escuro. No primeiro experimento para as variáveis comprimento médio da parte aérea, massa fresca da planta inteira, porcentagem de enraizamento e porcentagem de estacas brotadas houve decréscimo linear em função das concentrações de quitosana. No segundo experimento, houve efeito linear negativo em função das concentrações crescentes de quitosana, sendo que a inibição do crescimento micelial foi de 81,7 por cento, demonstrando o grande potencial do uso de quitosana no controle da antracnose da videira.

The objective of this research was to evaluate the effects of the chitosan on the development in vitro plantlets of grapevines cv. Merlot and its antifungal action on Elsinoe ampelina. In the first trial, explants of the grapevine cv. Merlot were transferred to growing medium DSD1 plus concentrations of 0; 25; 50; 100; 150 and 200 mg L-1 of the product. After 90 days of in vitro cultivation the plantlets were evaluated for root and leaf number, root and shoot length and fresh mass of roots and shoots. In the second experiment, doses of 0; 60; 120; 180; 240 and 300 mg L-1 were incorporated to the BDA media, where the fungus was inoculated. The mycelial diameter was evaluated at 6 and 9 days after incubation at 25° C in the dark. In the first experiment, for the variables shoot length average, plant fresh mass, rooting percentage and sprouted cuttings there was a linear reduction in function of chitosan doses. In the second trial, there was a negative linear effect with mycelial growth inhibition, of 81.7 percent, showing great the potential of chitosan in the control of grapevine anthracnose.

Garlic extract improves budbreak of the 'Niagara Rosada' grapevines on sub-tropical regions / Extrato de alho melhora a superação da dormência de videiras 'Niagara Rosada' em regiões subtropicais

In the regions with mild winters, usually the chilling requirements for bud dormancy release of grapevines are not fulfilled, and only the highly toxic products CaCN2 (calcium cyanamide) and H2CN2 (hydrogen cyanamide) are recommended in conventional viticulture for budbreak. The purpose of this study was to verify the effect of garlic extract on budbreak of 'Niagara Rosada' grapevines in two different vineyards. The highest dose of garlic extract (70mL L-1) showed a great potential for budbreak in organic production, presenting similar effects of cyanamides in budbreak. This treatment improved the sprouting percentage, number of clusters, accelerated the beginning of sprouting and reduced the cycle between pruning and harvest.

Nas regiões com invernos amenos, normalmente as exigências de frio para a quebra de dormência de gemas de videiras não são atendidas, e apenas os produtos altamente tóxicos CaCN2 e H2CN2 são recomendados na viticultura convencional para quebra de dormência. A proposta deste estudo foi verificar o efeito do extrato de alho na quebra de dormência de videiras 'Niagara Rosada', em dois diferentes vinhedos. A dose mais elevada de extrato de alho (70mL L-1) demonstrou ter um grande potencial para a quebra de dormência na produção orgânica, apresentando efeitos similares aos das cianamidas, na quebra de dormência. Este tratamento aumentou a porcentagem de brotação e o número de cachos, antecipou o começo da brotação e reduziu o ciclo entre poda e colheita.

Produção de antissoro policlonal utilizando a proteína capsidial recombinante do Rupestris stem pitting-associated virus / Production of polyclonal antiserum using recombinant coat protein of Rupestris stem pitting -associated virus

O Rupestris stem pitting-associated virus (RSPaV) é o agente causal das caneluras do lenho da videira. Este trabalho teve como objetivo produzir antissoro policlonal a partir da proteína capsidial (CP) recombinante do RSPaV e avaliar a sua especificidade e sensibilidade. O gene da CP do RSPaV, com 780pb, foi previamente caracterizado. Esse gene foi subclonado no sítio de restrição EcoRI, no vetor de expressão pRSET-B e o plasmídeo recombinante foi utilizado para induzir a expressão da CP em Escherichia coli. A CP, ligada a uma cauda de seis histidinas, foi purificada por meio de cromatografia de afinidade em coluna de Ni-NTA a partir do extrato de proteínas totais extraídas de E. coli. A identidade da proteína purificada foi confirmada em SDS-PAGE e Western blot, utilizando-se anticorpos comerciais contra a cauda de seis histidinas. A CP recombinante expressada in vitro apresentou massa molecular de cerca de 31kDa. A proteína purificada foi quantificada e 2,55mg foram utilizados para a imunização de um coelho. O antissoro policlonal obtido reagiu com diferentes isolados deste vírus, extraídos de videiras em ELISA indireto.

RSPaV is the causal agent of pitting in the grapevine woody cylinder. The aim of this research was to produce polyclonal antiserum against recombinant RSPaV coat protein (CP) and evaluate its specificity and sensibility. The CP gene (780bp) of RSPaV was previously characterized. This gene was subcloned into the EcoRI site of the pRSET-B expression vector and the recombinant plasmid was used to induce the expression of the CP in E. coli cells. The CP, fused to a 6-His-tag, was purified from E. coli total protein extract by affinity chromatography using Ni-NTA resin. Identity of the purified protein was confirmed by SDS-PAGE and Western blot, using antibodies against the histidine tail. The in vitro-expressed recombinant CP presented a MW of ca. 31kDa. The purified protein was quantified and 2.55mg used for the immunization of a rabbit. The obtained polyclonal antiserum reacted with different RSPaV isolates extracted from grapevines in indirect ELISA.

Genetic changes in grapevine genomes after stress induced by in vitro cultivation, thermotherapy and virus infection, as revealed by AFLP

The Amplification Fragment Length Polymorphism (AFLP) technique was employed to study genetic variations which can be induced in vines by the stress occurring during different aspects of viticulture (in vitro cultivation, in vitro thermotherapy and virus infection). Analysis of AFLP banding patterns, generated by using 15 primer combinations, pointed to negligible genetic variation among plants exposed to individual stress. The average of similarity coefficients between differently stressed plants of the cultivars Müller Thurgau and Riesling were 0.984 and 0.991, respectively, as revealed by AFLP analysis. The low incidence of observed polymorphism demonstrates the high level of genome uniformity in plants reproduced by in vitro micropropagation via nodes, those subjected to in vitro thermotherapy and virus-infected plants.

Efeito de concentrações de glicina e inositol no cultivo in vitro de duas frutíferas de clima temperado / Effect of glycine and inositol concentrations on two in vitro cultivated temperate fruit trees

A fim de aprimorar técnicas de cultivo in vitro de duas frutíferas de clima temperado (amoreira-preta cv. Tupy e porta-enxerto de videira cv. Kobber 5BB), testaram-se diferentes concentrações de glicina e inositol, adicionadas ao meio de cultura. Para plântula de amoreira-preta, o meio foi constituído do meio básico MS, acrescido de 30 g L-1 de sacarose e 7 g L-1 de ágar, e o pH ajustado para 5,8 antes da autoclavagem a 121ºC e 1 atm por 20 minutos; e do meio básico DSD1 para porta-enxerto de videira cv. Kobber 5BB, acrescido de 20 g L-1 de sacarose e 7 g L-1 de ágar, e o pH ajustado para 6,4. O experimento com amoreira-preta consistiu de 5 diferentes concentrações de glicina (0; 1,0; 2,0; 4,0 e 8,0 mg L-1), 5 de inositol (0; 50; 100; 200 e 400 mg L-1) e suas combinações. O experimento com videira consistiu de 4 diferentes concentrações de glicina (0; 1,0; 2,0 e 4,0 mg L-1), 4 de inositol (0; 10; 20 e 40 mg L-1) e suas combinações. Segmentos nodais, oriundos de plântulas preestabelecidas in vitro foram excisados e introduzidos em tubos de ensaio contendo 15 mL dos meios de cultura. Posteriormente, os tubos de ensaio foram transferidos para sala de crescimento a 25 ± 2ºC, irradiância de 32 mmol m² s¹ e fotoperíodo de 16 horas. O delineamento experimental utilizado foi o inteiramente casualisado, utilizando-se de doze plântulas por tratamento. Após 70 dias de cultivo in vitro, melhores resultados para a amoreira-preta cv. Tupy foram obtidos com concentração de glicina até a recomendada no meio de cultura MS (2,0 mg L-1) e 4 vezes o valor de inositol. Para o porta-enxerto de videira, melhores resultados foram obtidos na ausência e/ou com baixas concentrações de glicina e concentração de inositol igual ou superior à recomendada no meio de cultura DSD1.

Aiming to improve the in vitro cultivation techniques of two temperate fruit i.e. blackberry cv. Tupy and grapevine rootstock cv. Kobber 5BB, different glycine and inositol concentrations in the culture medium were tested. The culture medium was constituted of MS basal medium, added of 30 g L-1 sucrose and 7 g L-1 agar, and the pH adjusted to 5.8 before the sterilization of 121ºC and 1 atm for 20 minutes, and DSD1 basal medium for grapevine rootstock cv. Kobber 5BB, added of 20 g L-1 sucrose and 7 g L-1 agar, and the pH adjusted to 6.4. The work with blackberry consisted of 5 different concentrations of glycine (0; 1.0; 2.0; 4.0 and 8.0 mg L-1), 5 of inositol (0; 500; 100; 200 and 400 mg L-1) and its combinations. The work with grapevine composed of 4 different concentrations of glycine (0; 1.0; 2.0 and 4.0 mg L-1), 4 of inositol (0; 10; 20 and 40 mg L-1), and its combinations. Nodal segments from in vitro plants was excised and introduced into test tubes containing 15 mL of culture medium. After that, the culture tubes were transferred in a growth room to 25 ± 2ºC, irradiance of 32 mol m-2.s-1 and photoperiod of 16 hours. The experiments were settled in a completely randomized design, using twelve explants per treatment. After 70 days of in vitro cultivation, better results for the blackberry cv. Tupy were obtained with glycine concentration as recommended in the MS culture medium (2 mg L-1) and 4 fold of the inositol value. For the grapevine rootstock, better results were obtained in the absence and/or with low glycine concentrations and the same or higher inositol concentration as recommended in the DSD1 culture medium.

Ocorrência do ácaro vermelho europeu Panonychus ulmi (Koch) (Tetranychidae) associado à cultura da videira no Rio Grande do Sul, Brasil / European red spider mite Panonychus ulmi (Koch) (Tetranychidae) occurrence of vineyards in Rio Grande do Sul, Brazil

Neste artigo é descrita ocorrência e o dano causado pelo ácaro vermelho europeu Panonychus ulmi (Koch) associado à cultura da videira. As coletas foram realizadas na safra 2005-2006 em Vitis vinifera L. da cultivar Merlot, nos municípios de Bento Gonçalves e Candiota, no Estado do Rio Grande do Sul, Brasil. As folhas infestadas apresentaram bronzeamento, com manchas avermelhadas na face adaxial, resultando na queda prematura das mesmas. Esse é o primeiro registro do ácaro vermelho europeu danificando a cultura da videira no Rio Grande do Sul.

The occurrence and the damage caused for the European red spider mite Panonychus ulmi (Koch) associated to the culture of the grapevine are described. The collections had been carried through in 2005-2006 seasons in Vitis vinifera L. of cultivating Merlot in Bento Gonçalves and Candiota counties, in the state of Rio Grande do Sul, Brazil. The infested leaves had presented bronzing, with spots redly in the adaxial face resulting in the premature fall. This is the first register of the European red mite damaging the culture of the grapevine in Rio Grande do Sul.