A microfiltração tangencial aplicada à clarificação de vinho branco tem se tornado uma importante alternativa aos processos convencionais de filtração na indústria vinícola, como a filtração por terras diatomáceas, trasfega e centrifugação. O uso de membranas cerâmicas tem despertado interesse para esta finalidade. Neste trabalho, diferentes membranas cerâmicas e compósitas tubulares, mono e multicanal, de diferentes tamanhos de poro, foram avaliadas quanto à sua morfologia e testes de permeabilidade hidráulica e do vinho branco, a fim de verificar a influência da estrutura destas no fluxo de vinho branco permeado, bem como, avaliar a presença de resíduos sólidos suspensos no vinho permeado decorrente de seu processo de clarificação. As membranas foram caracterizadas por difração de raios X (DRX), microscopia eletrônica de varredura por emissão de campo (FESEM), espectroscopia de dispersão de energia (EDS), porosimetria por intrusão de mercúrio, testes de permeabilidade e retenção de proteínas. O tamanho médio de poro apresentado pelas membranas variou de 0,0056 a 5,0 µm. As membranas compósitas apresentaram ótima retenção proteica, 99%, para solutos com massa molar acima de 45 kDa. Membranas de α-alumina tratadas termicamente a 1450ºC e membranas compósitas de α-alumina/poliamida 66 demonstraram, em média, redução de 99,9% da turbidez inicial do vinho, sendo possível reduzir de 720 NTU para, em média, 0,30 NTU, com possibilidade de estabilização tartárica em sete dias e fluxos de permeado que variaram de 60,8 a 8,8 L.m-2.h-1, respectivamente, a 1,0 bar de pressão. Por outro lado, membranas de mulita e titânia não demonstraram eficiência para a clarificação, sendo que o vinho permeado pela membrana de mulita apresentou turbidez de 20 NTU e aumento do índice de polifenóis totais (IPT). Membranas compósitas permitiram a redução de 37% no valor de IPT do vinho branco. O baixo valor de turbidez obtido nos vinhos permeados pelas membranas de alumina e compósitas demonstra forte perspectiva da utilização destas membranas na clarificação do vinho branco. / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2017-03-24T18:24:14Z No. of bitstreams: 1 Tese Kélen Cristofoli.pdf: 5048002 bytes, checksum: 8a229e6f22e8e9bddc2c19f7a5060a08 (MD5) / Made available in DSpace on 2017-03-24T18:24:14Z (GMT). No. of bitstreams: 1 Tese Kélen Cristofoli.pdf: 5048002 bytes, checksum: 8a229e6f22e8e9bddc2c19f7a5060a08 (MD5) Previous issue date: 2017-03-24 / The tangential microfiltration applied to the clarification of white wine has become an important alternative to conventional filtration processes in the wine industry, for example, the filtration diatomaceous earth, land transfer and centrifugation. The use of ceramic membranes has awakened interest for this purpose. In this work, different and tubular composite ceramic membranes, mono and multi-channel, of different sizes of pore, were evaluated for their morphology and hydraulic permeability tests and white wine, in order to verify the influence of the structure of these in white wine flow permeated as well, evaluate the presence of suspended solids in the waste wine due to its process of clarification. The membranes were characterized by x-ray diffraction (XRD), scanning electron microscopy by field emission (FESEM), energy dispersive spectroscopy (EDS), porosimetria by mercury intrusion tests, permeability and protein retention. The average pore size presented by membranes ranged from 0.0056 to 5.0 µm. Composite membranes showed great protein retention, 99%, for solutes with molecular weight up to 45 kDa. α-alumina membranes treated thermally to 1450ºC and composite membranes of α-alumina/polyamide 66 showed, on average, 99.9% reduction of wine’s turbidity, being possible to reduce from 720 NTU to average 0.30 NTU, with possibility of tartaric stabilization in seven days and permeate fluxes that ranged from 60.8 to 8.8 L. m-2h-1, respectively, the 1.0 bar pressure. On the other hand, mullite and titania membranes have not shown effectiveness for clarification, the wine permeated by the membrane of mullite presented turbidity of 20 NTU and increase the rate of total polyphenols (IPT). Composite membranes have enabled the reduction of 37% in the value of IPT of the white wine. The low turbidity value obtained in wine permeated by composite and alumina membranes, strong perspective of use demonstrates of these membranes in the clarification of white wine.

A microfiltração tangencial aplicada à clarificação de vinho branco tem se tornado uma importante alternativa aos processos convencionais de filtração na indústria vinícola, como a filtração por terras diatomáceas, trasfega e centrifugação. O uso de membranas cerâmicas tem despertado interesse para esta finalidade. Neste trabalho, diferentes membranas cerâmicas e compósitas tubulares, mono e multicanal, de diferentes tamanhos de poro, foram avaliadas quanto à sua morfologia e testes de permeabilidade hidráulica e do vinho branco, a fim de verificar a influência da estrutura destas no fluxo de vinho branco permeado, bem como, avaliar a presença de resíduos sólidos suspensos no vinho permeado decorrente de seu processo de clarificação. As membranas foram caracterizadas por difração de raios X (DRX), microscopia eletrônica de varredura por emissão de campo (FESEM), espectroscopia de dispersão de energia (EDS), porosimetria por intrusão de mercúrio, testes de permeabilidade e retenção de proteínas. O tamanho médio de poro apresentado pelas membranas variou de 0,0056 a 5,0 µm. As membranas compósitas apresentaram ótima retenção proteica, 99%, para solutos com massa molar acima de 45 kDa. Membranas de α-alumina tratadas termicamente a 1450ºC e membranas compósitas de α-alumina/poliamida 66 demonstraram, em média, redução de 99,9% da turbidez inicial do vinho, sendo possível reduzir de 720 NTU para, em média, 0,30 NTU, com possibilidade de estabilização tartárica em sete dias e fluxos de permeado que variaram de 60,8 a 8,8 L.m-2.h-1, respectivamente, a 1,0 bar de pressão. Por outro lado, membranas de mulita e titânia não demonstraram eficiência para a clarificação, sendo que o vinho permeado pela membrana de mulita apresentou turbidez de 20 NTU e aumento do índice de polifenóis totais (IPT). Membranas compósitas permitiram a redução de 37% no valor de IPT do vinho branco. O baixo valor de turbidez obtido nos vinhos permeados pelas membranas de alumina e compósitas demonstra forte perspectiva da utilização destas membranas na clarificação do vinho branco. / The tangential microfiltration applied to the clarification of white wine has become an important alternative to conventional filtration processes in the wine industry, for example, the filtration diatomaceous earth, land transfer and centrifugation. The use of ceramic membranes has awakened interest for this purpose. In this work, different and tubular composite ceramic membranes, mono and multi-channel, of different sizes of pore, were evaluated for their morphology and hydraulic permeability tests and white wine, in order to verify the influence of the structure of these in white wine flow permeated as well, evaluate the presence of suspended solids in the waste wine due to its process of clarification. The membranes were characterized by x-ray diffraction (XRD), scanning electron microscopy by field emission (FESEM), energy dispersive spectroscopy (EDS), porosimetria by mercury intrusion tests, permeability and protein retention. The average pore size presented by membranes ranged from 0.0056 to 5.0 µm. Composite membranes showed great protein retention, 99%, for solutes with molecular weight up to 45 kDa. α-alumina membranes treated thermally to 1450ºC and composite membranes of α-alumina/polyamide 66 showed, on average, 99.9% reduction of wine’s turbidity, being possible to reduce from 720 NTU to average 0.30 NTU, with possibility of tartaric stabilization in seven days and permeate fluxes that ranged from 60.8 to 8.8 L. m-2h-1, respectively, the 1.0 bar pressure. On the other hand, mullite and titania membranes have not shown effectiveness for clarification, the wine permeated by the membrane of mullite presented turbidity of 20 NTU and increase the rate of total polyphenols (IPT). Composite membranes have enabled the reduction of 37% in the value of IPT of the white wine. The low turbidity value obtained in wine permeated by composite and alumina membranes, strong perspective of use demonstrates of these membranes in the clarification of white wine.

This thesis is focused on the problems of the prediction of financial situation of non-financial companies in the Czech Republic and the European Union with shares publically traded on the stock exchange. It is based on the original accounting and structural models of credit risk which enable to enumerate the probability of default. The application of Merton, Altman and Ohlson model in their basic theoretical form for the determination of the probability of default or bankruptcy is dealt with in detail. Modern multi-dimensional econometric and simulation approaches enabled to extend the original Merton model, adding the theory of multi-dimensional D-Vine copulas which project the dependency between the development on the financial market into the structure of dependency of company assets of companies, which serve as a significant indicator for the calculation of the probability of default. Next steps proceed from the creation of prediction models based on the SVM method and decision trees for the purpose of classification of company financial difficulties which are represented by the debt index or liquidity index. Different forms and numbers of input variables are used for this purpose: accounting, market and combined data together with the derived probabilities of default, or more precisely of financial distress.

The reflection of the Anglo-American institute of the agreement on guilt and punishment in the chosen European countries Abstract The goal of this thesis is to describe which elements of an Anglo-American institute of the agreement on guilt and punishment (represented by the American federal legal regulation as the most influential regulation of this institute) are reflected in legal regulations of an agreement on guilt and punishment in chosen European countries (Slovakia and the Czech Republic) by a detailed analysis of particular legal regulations. This thesis is supposed to inform about the history of the institute of the agreement on guilt and punishment in countries of its origin (England, Wales and the USA) including its spreading into countries with European continental law tradition. The thesis is systematically divided into four chapters. The first chapter discusses the history of the institute of the agreement on guilt and punishment and individual subchapters describe historical development in the USA, historical development in England and Wales including its spreading into countries with European continental law tradition, the author also gives reasons why this is happening. The second chapter is focused on defining of key concepts of the thesis which are 'plea bargaining', the agreement on...

京都大学 / 新制・課程博士 / 博士(農学) / 甲第24659号 / 農博第2542号 / 新制||農||1098(附属図書館) / 学位論文||R5||N5440(農学部図書室) / 京都大学大学院農学研究科森林科学専攻 / (主査)教授 北山 兼弘, 教授 北島 薫, 教授 柴田 昌三 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

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Organic mulches and cover crops mulched in situ were assessed for their effects on B. cinerea primary inoculum and disease levels in inflorescences at flowering and/or bunches at harvest. Organic mulches were used to enhance biological degradation of vine debris to reduce levels of B. cinerea primary inoculum the following season. Four mulch types (anaerobically and aerobically fermented marc (grape pressings), inter-row grass clippings and shredded office paper) were applied under ten-year-old Riesling vines in a ten-replicate randomized block design in New Zealand over two consecutive years. Plastic mesh bags, each containing naturally infected vine debris, were placed under vines on bare ground (control) and at the soil-mulch interface, in winter (July) 2003 and 2004. In each year, half the bags were recovered at flowering (December) and the remainder at leaf plucking (February), for assessment of B. cinerea sporulation from the vine debris and debris degradation rate. Bait lamina probes, which measure soil biological activity, were placed in the soil-mulch interface three weeks before each of the two bag-recovery dates in both years and were then removed and assessed at the same times as were the bags. All mulches led to a reduction in B. cinerea sporulation. This reduction was significantly correlated with elevated rates of vine debris decomposition and increased soil biological activity. Over both years, compared with the controls, all treatments gave a 3-20-fold reduction in B. cinerea sporulation, a 1.6-2.6-fold increase in vine debris degradation and in the two marc and the paper treatments, a 1.8-4-fold increase in activity of soil organisms. The mulches also altered vine characteristics and elevated their resistance to B. cinerea through changes to the soil environment. Functional soil biological activity, as measured by Biolog Ecoplates and bait lamina probes, was increased 2-4 times in the two marc and paper treatments, compared with the control, an effect relating to the elevated soil moisture and reduced temperature fluctuations under these mulches. Soil nutrient levels and the C:N ratios were also affected in these treatments. The mulched paper lowered vine canopy density by up to 1.4 times that of the other treatments, an effect which probably led to elevated light penetration into the canopy and consequent increased canopy temperature, photosynthesis and lowered canopy humidity. These changes to soil and vine characteristics increased grape skin strength by up to 10% in the paper treatment and sugar concentrations by 1.2-1.4 °Brix in the two marc and paper treatments. The severity of B. cinerea infections in the anaerobic marc, aerobic marc and paper treatments were reduced to 12%, 3% and 2.2% of the control, respectively, in field assessments averaged over two consecutive harvests. Cover crops mulched in situ had similar effects to those of the organic mulches, increasing soil biological activity and reducing B. cinerea primary inoculum and the severity of B. cinerea infection in grapes at harvest (2006). Inter-row phacelia and ryegrass were mulched in winter 2005 and compared with a bare ground control, under 10-year-old Chardonnay vines in a ten-replicate randomized block design. Functional soil biological activity increased by 1.5-4.5 times in the two cover crop treatments compared with the control, an effect possibly related to elevated soil moisture in these treatments. This increase in soil moisture and soil biological activity increased vine debris degradation, reduced B. cinerea primary inoculum on the debris and decreased B. cinerea severity at flowering (December 2005) and harvest (April 2006). These results show the potential of organic mulches and cover crops mulched in situ to enhance soil ecosystem services and improve the sustainability of viticultural practices.

Thesis (PhD (Conservation Ecology and Entomology))--Stellenbosch University, 2008. / The mutual association between honeydew foraging ants and vine mealybugs in vineyards is detrimental to the biological control of the vine mealybug Planococcus ficus (Signoret). This study investigated the relationship between ants, vine mealybugs and their parasitoids to improve biological control of the vine mealybug. The investigation was carried out during two consecutive growing seasons (2005-2006 and 2006-2007) in two major wine grape growing areas of the Western Cape Province, Simondium (two farms) and Breede River Valley (one farm). A presence – absence cluster sampling system was used on a biweekly basis throughout both growing seasons from October to February on each of the farms. Additionally, yellow delta traps with vine mealybug pheromone, were used to catch parasitoids in vineyards. Prior to harvest, economic damage to grape bunches was assessed using a 0-3 damage rating index. Three ant species, Anoplolepis steingroeveri (Forel), Crematogaster peringueyi Emery and Linepithema humile (Mayr) foraged mainly on vine stems during both seasons, relative to other plant parts. Vine mealybugs of all stages were found on all sampling dates. Three species of primary parasitoids attacked the vine mealybug, including Anagyrus sp. near pseudococci (Girault), Coccidoxenoides perminutus (Timberlake) and Leptomastix dactylopii Howard. Bunch damage was significantly different between seasons in the Breede River Valley but not in Simondium. There was a negative linear relationship between ants and parasitoid activity in all vineyards. Spatial Analysis by Distance IndicEs (SADIE) was used to analyse spatial distribution of ants and parasitoids and ArcView, with its extension, Spatial Analyst, were used to map the gap, patch and local association indices where significant association and disassociation occurred. A significant association was found between C. peringueyi and parasitoids and L. humile and parasitoids. There was a significant disassociation between L. humile and C. peringueyi and between A. steingroeveri and C. peringueyi indicating interspecific hostility.

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae), the vine mealybug, is of economic importance to the wine and table grape (viticulture) industries, as it characteristically causes more damage than other mealybug species. Mealybug infestations contaminate grapes with their waxy secretions, egg-sacs and honeydew production, on which sooty mould grows, resulting in the fruit being unmarketable. Many export grapes are rejected, prior to shipment, as a result of infestations and phytosanitary concerns with regard to mealybug infestations. They are also vectors for various plant viruses. Up to date, the most common method of mealybug control in South Africa has been the use of chemical insecticides. Unfortunately, mealybugs are difficult to control chemically, due to their secretive/hidden lifestyle, where chemicals do not reach them. Of great concern is the ability of mealybugs to rapidly build up resistance to insecticides as well as the negative environmental effects associated with chemical pesticide use. Alternatively, entomopathogenic nematodes (EPNs), belonging to the families Heterorhabditidae and Steinernematidae, have been identified as lethal insect pathogens and their insecticidal action, towards a variety of insect pests, has proven them to be valuable and effective biocontrol agents. Laboratory bio-assays, to determine the ability of eight different EPN isolates to infect and kill P. ficus, were conducted. Six of the isolates were indigenous species and the other two, Heterorhabditis bacteriophora and Steinernema feltiae, were produced in Germany and are commercially available in South Africa. Planococcus ficus was highly susceptible to two indigenous species, Heterorhabditis zealandica and Steinernema yirgalemense; responsible for 96% ± 2% and 65% ± 10% mealybug mortalities, respectively. Biological studies illustrated that both H. zealandica and S. yirgalemense are able to complete their life cycles within adult female P. ficus. There was no significant difference in the pathogenicity of commercially produced H. bacteriophora, recycled through an insect host, and those from the formulated commercial product. However, commercially produced S. feltiae individuals, that were recycled through an insect host, were statistically more effective than those that were not. The LC50 and LC90 values for H. zealandica, in the current study, were 19 and 82 infective juveniles (IJs) respectively, which were similar to the LC50 and LC90 values for S. yirgalemense at 13 and 80, respectively. The LC50 and LC90, for commercially available H. bacteriophora, were greater than they were for both H. zealandica and S. yirgalemense, with values of 36 and 555, respectively. Such results indicate that there is a definite positive relationship which exists between the concentration of IJs of all three nematode species, used for inoculation, and the percentage mortality of P. ficus. Sand column tests resulted in S. yirgalemense outperforming H. zealandica significantly, with average mortalities of 95% ± 1.4% and 82% ± 4.1%, respectively. As a result S. yirgalemense was chosen for further studies in the field. IJs of commercially produced H. bacteriophora and S. feltiae were exposed to imidacloprid in laboratory bioassays to determine the effect on survival and infectivity. This study established the fact that these two EPN species can be applied, in combination with imidacloprid, in an integrated pest management scheme. Soil application field trials at Welgevallen and Nietvoorbij, using S. yirgalemense and mealybugs in Eppendorf tubes, buried 15 cm in the soil, resulted in 50% ± 10% and 52% ± 12% mealybug mortalities, respectively, when applying IJs at a concentration of 80 IJs/cm2. No significant difference was found between mealybug mortalities as a result of the three IJ concentrations applied (20, 40 and 80 IJs/cm2) for both vineyards. Persistence trials indicated that after four months post application, Cydia pomonella larval mortalities showed no significant reduction in infectivity on the Welgevallen vineyard, while on the Nietvoorbij vineyard there were no larval mortalities. Tests to establish whether or not S. yirgalemense and H. zealandica produced ant deterrent factors, showed no significant differences between the number of intact cadavers for both nematode species and for cadavers that were either four or six days old. There is, however, indication that deterrent factors may be in action in cadavers that were used six days after inoculation with 60% and 49% remaining intact for H. zealandica and S. yirgalemense infected cadavers respectively. All freeze killed cadavers were consumed by Linepithema humile (Mayr) (Argentine ant). The effects of low temperatures on EPN movement and infectivity were tested for H. zealandica and S. yirgalemense in the laboratory. The mortality of P. ficus at 14˚C, as opposed to 25˚C, for S. yirgalemense and H. zealandica were found to be 9.1% ± 2.6% and 2.5% ± 1.2% respectively. Vertical sand column tests were also conducted at 14˚C for S. yirgalemense and H. zealandica, which produced low mealybug mortalities of 3.5% ± 2.4% and 8.5% ± 1.4% respectively. This illustrates the low infectivity of the two local species at low temperatures. Laboratory persistence trials, conducted over a period of four months with S. yirgalemense, showed steady persistence of 100%, while H. zealandica had a statistically significant decrease of codling moth mortalities to 44% ± 5%. A three armed olfactometer was designed to establish if S. yirgalemense responds and moves towards chemical cues in the soil. A significant greater average number of IJs moved towards the grape vine roots (246 ± 0.124 IJs), than to the mealybugs (133 ± 0.168 IJs) and to the control (4 ± 1.02 IJs). This demonstrates that S. yirgalemense does actively seek out its hosts and that volatile cues produced by damaged grape vine roots, are more attractive to the EPN than cues produced by P. ficus. This study illustrates that S. yirgalemense has great potential as a biopesticide for controlling P. ficus in the soil of South African grape vineyards. Emphasis was placed on soil application of S. yirgalemense in the field, which produced good results, while laboratory tests indicate the potential for further aerial field application trials on grape vines. As the EPNs are not negatively affected by the agrochemical imidacloprid, the simultaneous use and combined action of both agents will potentially provide the farmer with excellent control against P. ficus. Further field- and aerial application studies will complement the current study and hopefully provide positive results for the efficient control of P. ficus found on grape vines. / AFRIKAANSE OPSOMMING: Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae), die wingerd witluis, is van groot ekonomiese belang vir die wyn en tafeldruif industrieë, aangesien dit kenmerkend meer skade veroorsaak as enige ander witluis spesies. Witluis infestasies besmet druiwe met hulle wasagtige afskeidings, eierssakke en heuningdou produksie, waarop swamme groei, wat tot gevolg het dat die druiwe onbemarkbaar is. Baie besendings druiwe, bestem vir uitvoer, word afgekeur weens witluis besmettings en ook as gevolg van fitosanitêre oorwegings. Hulle tree ook op as vektore van verskeie plantvirusse. Die mees algemene manier waarmee witluis in Suid-Afrika beheer word, is chemiese behandeling. Ongelukkig is witluis baie moeilik om chemies te beheer vanweë hulle verskuilde lewenswyse wat dit moeilik maak vir chemikalieë om hulle te bereik. Die vermoë van witluis om vinnig weerstand op te bou teen insekdoders, asook die negatiewe effek van chemiese middels op die omgewing, is kommerwekkend. Alternatiewelik, kan entomopatogeniese nematodes (EPNs) van die families Heterorhabditidae en Steinernematidae gebruik word vir die beheer van witluis. Hierdie nematodes is geïdentifiseer is as dodelike insek patogene, vir ʼn groot verskeidenheid van pes insekte en daar is bewys dat hulle as waardevolle en effektiewe biologiese beheer agente kan optree. Laboratorium biotoetse is gedoen om die vermoë van agt EPN isolate te evalueer om P. ficus te beheer. Ses van die EPN isolate is inheems, terwyl die ander twee, Heterorhabditis bacteriophora en Steinernema feltiae, in Duitsland produseer is en kommersieel beskikbaar is in Suid-Afrika. Planococcus ficus is hoogs vatbaar vir die twee inheemse EPN spesies, naamlik Heterorhabditis zealandica en Steinernema yirgalemense en hulle is verantwoordelik vir 96% ± 2% en 65% ± 10% van witluis mortaliteit. Biologiese studies het aangetoon dat beide H. zealandica en S. yirgalemense in staat is om hul lewensiklus te voltooi in volwasse wyfies van P. ficus. Daar is geen beduidende verskil gevind in die patogenisiteit van die geformuleerde kommersiële produk H. bacteriophora en dié wat in vivo geproduseer is nie. Daar is egter in die geval van S. feltiae, gevind dat die nematodes, wat in insekte produseer is, statisties beduidend meer effektief was, as dié wat kommersieel beskikbaar was. Die LC50 en die LC90 waardes van H. zealandica, in die huidige studie, was 19 en 82 infektiewe larwes (IJs), wat baie naby die LC50 en LC90 waarders van S. yirgalemense van 13 en 80 was. Die LC50 en LC90 vir die kommersieel beskikbare H. bacteriophora was groter as vir beide H. zealandica en S. yirgalemense, met waardes van 36 en 555 onderskeidelik. Hierdie resultate dui daarop dat daar ʼn positiewe verwantskap bestaan tussen die konsentrasie van die IJs van drie EPN spesies en die persentasie mortaliteit van P. ficus. Sand kolomtoetse dui daarop dat S. yirgalemense baie beter vaar as H. zealandica met gemiddelde mortaliteite van 95% ± 1.4% en 82% ± 4.1% onderskeidelik. Op grond van hierdie resultate is S. yirgalemense gebruik vir verdere veld studies. IJs van kommersieel geproduseerde H. bacteriophora en S. feltiae is in laboratorium biotoetse blootgestel aan imidacloprid om die effek op die oorlewing en infektiewe vermoë vas te stel. Hierdie studie het aangetoon dat die twee EPN spesies aangewend kan word saam met imidacloprid in ʼn geïntegreerde plaagbestuur opset. Grond aanwendings is in veld proewe by Welgevallen en Nietvoorbij gedoen deur gebruik te maak van S. yirgalemense en P. ficus volwasse wyfies in Eppendorf buisies, 15 cm in die grond begrawe, het albei 50% ± 10% en 52% ± 10% witluis mortaliteit, respektiewelik, tot gevolg gehad, met die toediening van nematodes teen ʼn konsentrasie van 80 IJs/cm2. Geen beduidende verskille is gevind tussen die witluismortaliteit en die resultate van die verskillende EPN konsentrasies (20, 40 en 80 IJs/cm2) wat op beide wingerde toegedien is nie. Oorlewings toetse het aangedui dat, drie maande na toediening, met Cydia pomonella as indikator, geen beduidende verskille in die infeksie potensiaal van die Welgevallen wingerd to gevolg gehad het nie, terwyl daar op die Nietvoorbij wingerd geen verdere larvale mortaliteit gevind is was nie. Toetse om vas te stel of S. yirgalemense en H. zealandica afkrikmiddels vir miere in besmette insek kadawers produseer het aangetoon dat daar geen beduidende verskil is tussen die getal kadawers wat intakt is vir beide EPN spesies en kadawers wat vier of ses dae oud is nie. Daar is egter aangetoon dat die afskrikmiddels wel ses dae na infeksie deur insek kadawers afgeskei word; aangesien 60% en 49% van die oorblywende kadawers nog volledig was toe dit geïnfekteer was met H. zealandica en S. yirgalemense, onderskeidelik. Al die insek kadawers, wat deur bevriesing doodgemaak is, was deur Linepithema humile (Mayr) (Argentynse mier) verorber.

Dans cette étude, nous avons dans un premier temps adapte le protocole d'embryogenèse somatique primaire a différentes variétés d'hybrides porte-greffes (3309C, 110R, Fercal, 41B et SO4) en nous appuyant sur l'expérience acquise au laboratoire sur Vitis vinifera cv Chardonnay. Les résultats montrent que le génotype, le type d'explant (étamine, fleur ou nœud), le type et la dose d'auxine utilisés dans le milieu d’induction (2,4-D ou 2,4,5-T) ont une influence sur les efficacités d'embryogenèse somatique. En effet, pour le 3309C, l'utilisation du 2,4,5-T dans le milieu d'induction a montré une efficacité embryogène supérieure à partir de nœuds par rapport à celle obtenue à partir d'étamines. Cependant la meilleure efficacité a été obtenue à partir de fleurs de cette variété, sur un milieu d'induction contenant du 2,4-D. De plus, le protocole d'embryogenèse somatique secondaire utilise de manière récurrente au laboratoire nous a permis d'obtenir des masses embryogènes ainsi que des embryons somatiques secondaires de ces porte-greffes. Le protocole de conversion des embryons en plantes, en présence de 4,5 uM de cytokinine (BAP) s'est avère efficace pour le 11OR et le 41B. Dans un second temps, nous avons co-cultivé le matériel embryogène obtenu pour quatre de ces génotypes (110R, 3309C, Fercal et 41B), avec Agrobacterium tumefaciens contenant trois constructions génétiques : (i) une copie d'une séquence partielle (1020 pb) du gène de la coque protéique du virus en orientation sens; (ii) une partie courte en sens et en anti-sens (280 pb) de cette même séquence formant une structure en épingle a cheveux (hpRNA = hairpin RNA) ; (iii) un amiRNA ciblant une séquence virale. Le gène bactérien codant la néomycine phosphotransférase et conférant la résistance à un antibiotique, la kanamycine, a été utilisé comme gène de sélection. Les conditions de sélection a la kanamycine ont nécessité des adaptations expérimentales telles que l’ajustement de la concentration en antibiotique puisque la sélection avec 75 mg.L-1 de kanamycine s'avère insuffisamment drastique dans Ia plupart de nos expériences de co-cullture. Les résultats d'analyse moléculaire par PCR ont montré l'amplification probable des fragments d'intérêt (CPGFLV et amiRI1TA-71) dans des échantillons de 11OR et de 41B résistants à la kanamycine. Cependant des analyses moléculaires supplémentaires par AL-PCR ne nous ont pas renseignées sur une éventuelle intégration du transgène amiRATA-71 dans des masses embryogènes de 41B. / In this study, we initially adapted the protocol of primary somatic embryogenesis in different varieties of hybrid rootstocks (3309C, 110R, Fercal, 41B and SO4) building on the experience gained in the laboratory on Vitis vinifera cv Chardonnay. The results show that the genotype, the explant type (stamen, flower or node), the type and the dose of auxin used in the induction medium (2,4-D or 2,4,5-T) influence the efficiency of somatic embryogenesis. Indeed, for the 3309C, the use of 2,4,5-T in the induction medium showed a higher efficiency from embryogenic nodes compared to that obtained from stamens. However, the better efficiency was obtained from the flowers of this variety on an induction medium containing 2,4-D. In addition, a protocol used in the laboratory for secondary somatic embryogenesis allowed us to obtain embryogenic masses as well as secondary somatic embryos from these rootstocks. The protocol conversion of embryos into plants, in the presence of 4.5 [tM of cytokinin (BAP), was effective for the 110R and 41B. In a second step, we co-cultivated embryogenic material obtained for four of these genotypes (110R, 3309C, Fercal and 41B), with Agrobacteriwn tumefaciens containing three genetic constructs: (i) a copy of a partial sequence (1020 bp) of the coat protein gene of the virus in the sense orientation, (ii) a short part-way and antisense (280 bp) of the same sequence forming a hairpin structure (hairpin RNA = hpRNA) (iii) one amiRNA targeting a viral sequence. The nptll bacterial gene encoding neomycin phosphotransferase and conferring resistance to the antibiotic kanamycin, was used as the selection gene. The selection conditions to kanamycin have required experimental adaptations such as adjusting the concentration of antibiotic because the selection with 75 mg.L-1 of kanamycin was not enough drastic in most of our experiments of co-culture. The results of molecular analysis by PCR showed probable amplification of fragments of interest (CPGFLV and amiRNA-71) in samples of 11OR and 41B resistant to kanamycin. However, additional molecular analysis by AL-PCR did not inform us about a possible integration of the transgene amiRNA-71 in embryogenic masses of 41B.