A Escaldadura das Folhas da Ameixeira é o principal problema fitossanitário para a produção de ameixas no Brasil. A doença é causada pela bactéria Xylella fastidiosa, transmitida por cigarrinhas (Hemiptera: Cicadellidae, Cicadellinae). Neste trabalho foi realizado o levantamento, análise faunística e a flutuação populacional de Cicadellinae em pomares de ameixeira localizados nos Estados do Rio Grande do Sul (RS) e São Paulo (SP). A captura das cigarrinhas foi realizada com cartões adesivos amarelos (8,5 x 11,5 cm) em dois pomares de ameixeira de cada estado, localizados nos municípios de Paranapanema (SP) e Bento Gonçalves e Farroupilha (RS). Em cada pomar, foram instalados 20 cartões, distribuídos em 10 pontos espaçados 35 x 35 m, com duas alturas de amostragem (0,5 e 1,7m acima do nível do solo). Os cartões adesivos foram trocados quinzenalmente no período de setembro de 2006 a setembro de 2008. Com base na análise faunística, as espécies de cigarrinhas mais abundantes, constantes, freqüentes e dominantes foram analisadas para se conhecer a sua flutuação populacional. Nos dois pomares do RS foram coletados 1103 indivíduos de Cicadellinae distribuídos em 27 espécies. Destas, Erythrogonia dorsalis (Signoret, 1853), Sibovia sagata (Signoret,1854), Spinagonalia rubrovittata Cavichioli, 2008, Macugonalia cavifrons Stål, 1862, Dilobopterus dispar (Germar, 1821), Bucephalogonia xanthophis (Berg, 1879) e Molomea lineiceps Young, 1968 foram predominantes. Em SP, foram coletados 293 cicadelíneos, distribuídos em 10 espécies, sendo Oncometopia facialis (Signoret, 1854) e Molomea consolida Schröder, 1959 predominantes. Verificou-se uma menor diversidade e abundância de espécies de Cicadellinae nos pomares localizados em SP quando comparados com os pomares do RS. A distribuição populacional das espécies predominantes nos pomares do RS concentrou-se nos meses de janeiro a março, e em SP, de outubro a janeiro, correspondendo aos períodos de colheita e póscolheita em ambas as regiões. Os menores índices de captura de cicadelíneos foram obtidos nos meses de entressafra em todos os pomares amostrados. / The plum leaf scald (PLF) is a major threat for plum production in Brazil. PLF is caused by the bacteria Xylella fastidiosa which is transmitted by sharpshooter leafhoppers (Hemiptera: Cicadellidae, Cicadellinae). In this investigation, we surveyed the Cicadellinae species associated with plum orchards located in Rio Grande do Sul (RS) and São Paulo (SP) states, Brazil, in order to identify potential vectors of X. fastidiosa in these regions. We also studied the seasonal fluctuation of predominant species determined by faunistic analysis. In each area, adult sharpshooters were sampled using yellow sticky cards (8,5 x 11,5 cm) in two orchards in Paranapanema (SP) and one in Bento Gonçalves and Farroupilha (RS), respectivelly. In all orchards, traps were spaced 35 m apart in 10 sample units. Vertical distribution was also evaluated by placing cards at 0.5 and 1.7 m above soil level in each sample unit. The survey was conducted during two years from September 2006 to September 2008, by removing sticky cards fortnightly. In plum orchards located in RS state, we collected a total of 1103 specimens of 27 sharpshooter species. Erythrogonia dorsalis (Signoret, 1853), Sibovia sagata (Signoret,1854), Spinagonalia rubrovittata Cavichioli, 2008, Macugonalia cavifrons Stål, 1862, Dilobopterus dispar (Germar, 1821), Bucephalogonia xanthophis (Berg, 1879) and Molomea lineiceps Young, 1968 were predominant in these orchards. We collected 293 individuals of 10 sharpshooter species in SP state, where Oncometopia facialis (Signoret, 1854) and Molomea consolida Schröder, 1959 were predominant. We found a lower diversity and abundance of Cicadellinae species in plum orchards located in SP when compared to RS state. The predominant species were more abundant from January to March in RS and, from October to January in SP, corresponding to plum harvest and pos-harvest periods in both regions. The lowest population of sharpshooters in plum orchards was observed during the plant dormancy period.

A Escaldadura das Folhas da Ameixeira (EFA) é uma das principais doenças que prejudicam a produção de ameixas no Brasil. Ela é causada pela bactéria Xylella fastidiosa (Wells) cujos potenciais vetores são cigarrinhas (Hemiptera: Cercopidae e Cicadellidae, Cicadellinae). Sabe-se que existem diversas espécies de cicadelídeos e cercopídeos em pomares de ameixeira, mas faltam informações sobre as plantas hospedeiras desses insetos e sua importância epidemiológica. Esta pesquisa teve por objetivo associar as cigarrinhas potenciais vetoras com as plantas de ameixeira e com plantas infestantes da vegetação de cobertura dos pomares. Para tal, realizaram-se amostragens de cigarrinhas em três pomares de ameixeira no município de Paranapanema-SP, no período de setembro/2012 a abril/2013, usando-se três métodos distintos: a) rede de varredura em plantas infestantes; b) armadilhas adesivas amarelas colocadas na copa das ameixeiras a 0,5 e 2 m acima do solo; e c) amostragens visuais em ameixeiras e certas plantas infestantes. As cigarrinhas coletadas foram triadas e identificadas em laboratório e os resultados obtidos foram submetidos à análise faunística. Para verificar se as plantas infestantes eram hospedeiras da X. fastidiosa, experimentos de inoculação mecânica foram feitos para tentar estabelecer infecção pela bactéria nas plantas de Bidens pilosa L., Parthenium hysterophorus L., Raphanus sativus L., Euphorbia heterophylla L., Sida rhombifolia L., Solanum americanum Mill. e Lantana camara L. Após meses da inoculação, as plantas foram testadas por PCR e isolamento primários para detectar a infecção por X. fastidiosa. Avaliou-se, também, a ocorrência de transmissão de X. fastidiosa de ameixeiras paras plantas infestantes, por cigarrinhas sabidamente vetoras, Sibovia sagata (Signoret) e Macugonalia cavifrons (Stål). Nas amostragens com rede de varredura, encontraram-se 72 espécies de cigarrinhas associadas às plantas infestantes dos pomares de ameixeiras, pertencentes às famílias Achilidae, Cercopidae, Cicadellidae, Delphacidae, Derbidae, Dictyopharidae, Flatidae e Membracidae. As cigarrinhas foram observadas em um total de oito espécies herbáceas de dicotiledôneas e sete monocotiledôneas. As plantas infestantes que abrigam maiores números de cigarrinha são Paspalum notatum Flügge, Parthenium hysterophorus L. e Raphanus sativus L. Dentre as espécies com potencial de transmitir X. fastidiosa, o cercopídeo Deois schach (Fabricius) e o cicadelíneo Plesiommata corniculata Young predominaram nas plantas infestantes dos pomares de ameixeira, podendo ter um papel chave em uma eventual disseminação primária de X. fastidiosa dessas plantas para ameixeira. Os cicadelíneos Acrogonia citrina Marucci & Cavichioli e Oncometopia facialis (Signoret) predominaram em capturas com armadilhas adesivas amarelas na copa das ameixeiras, o que sugere sua participação na disseminação secundária de X. fastidiosa entre árvores de ameixeira. O. facialis foi visualizada nos ramos de ameixeiras e de Lantana camara L. As plantas infestantes Solanum americanum Mill e L. camara permitem colonização por X. fastidiosa após inoculação mecânica. A cigarrinha Sibovia sagata (Signoret) é capaz de transmitir X. fastidiosa de ameixeira para S. americanum. / Plum Leaf Scald (PLS) is one of the major diseases that impair the production of plums in Brazil, caused by the xylem-limited bacterium Xylella fastidiosa (Wells), whose potential vectors in plums are sharpshooter leafhoppers (Hemiptera: Cicadellidae, Cicadellinae) and spittlebugs (Hemiptera: Cercopidae). A number of leafhoppers and and spittlebugs have been reported in Brazilian plum orchards, but their host plants and role in PLS epidemiology are largely unknown. The goal of this research was to investigate the association of potential hopper vectors with plum trees and weedy plants in the ground vegetation of orchards, in order to determine key vector species and weeds involved in PLS epidemiology. Therefore, a hopper survey was carried out in three plum orchards in the municipality of Paranapanema, SP, from September/2012 to April/2013, using three sampling methods: a) sweep net on weed species of the ground vegetation; b) yellow sticky cards placed on the plum canopy at 0.5 and 2 m above ground; and c) visual inspections of plum trees and some weeds. The collected hoppers were sorted and identified in the laboratory, and the data were submitted to faunistic analysis. To check if the weeds were hosts of plum strains of X. fastidiosa, bacterial suspensions were mechanically inoculated in Bidens pilosaL., Parthenium hysterophorus L., Raphanus sativus L., Euphorbia heterophylla L., Sida rhombifolia L., Solanum americanum Mill. Lantana camara L. The plants were assayed for infection of X. fastidiosa by PCR and culture at 2 months after inoculation. Transmission assays of X. fastidiosa from plum to weeds were carried out using two sharpshooter vectors, Sibovia sagata (Signoret) e Macugonalia cavifrons (Stål). The sweep net samplings revealed 72 species of seven hopper families (Achilidae, Cercopidae, Cicadellidae, Delphacidae, Derbidae, Dictyopharidae, Flatidae and Membracidae) associated with eight dicotyledoneous and seven monocotyledoneous weeds in the ground vegetation, with prevalence of Cicadellidae and Cercopidae. Among the potential hopper vectors, the spittlebug Deois schach (Fabricius) and the sharpshooter Plesiommata corniculata Young were predomimant species on the weed species, suggesting that they may play a key role in a possible primary spread of X. fastidiosa from weeds to plum. Paspalum notatum Flügge, Parthenium hysterophorus L. and Raphanus sativus L. were the weed species with the largest hopper populations. The sharpshooters Acrogonia citrina Marucci & Cavichioli e Oncometopia facialis (Signoret) were prevalent species trapped by the yellow sticky cards on the plum canopy, indicating that they may be involved in secondary spread of X. fastidiosa between plum trees in that region. O. facialis was visually detected on branches of plum trees and on the weed Lantana camara L. The weeds Solanum americanum Mill e L. camara allow colonization by X. fastidiosa after mechanical inoculation.The sharpshooter Sibovia sagata (Signoret) transmitted X. fastidiosa from plum to S. americanum.

A Escaldadura das Folhas da Ameixeira (EFA) é uma das principais doenças que prejudicam a produção de ameixas no Brasil. Ela é causada pela bactéria Xylella fastidiosa (Wells) cujos potenciais vetores são cigarrinhas (Hemiptera: Cercopidae e Cicadellidae, Cicadellinae). Sabe-se que existem diversas espécies de cicadelídeos e cercopídeos em pomares de ameixeira, mas faltam informações sobre as plantas hospedeiras desses insetos e sua importância epidemiológica. Esta pesquisa teve por objetivo associar as cigarrinhas potenciais vetoras com as plantas de ameixeira e com plantas infestantes da vegetação de cobertura dos pomares. Para tal, realizaram-se amostragens de cigarrinhas em três pomares de ameixeira no município de Paranapanema-SP, no período de setembro/2012 a abril/2013, usando-se três métodos distintos: a) rede de varredura em plantas infestantes; b) armadilhas adesivas amarelas colocadas na copa das ameixeiras a 0,5 e 2 m acima do solo; e c) amostragens visuais em ameixeiras e certas plantas infestantes. As cigarrinhas coletadas foram triadas e identificadas em laboratório e os resultados obtidos foram submetidos à análise faunística. Para verificar se as plantas infestantes eram hospedeiras da X. fastidiosa, experimentos de inoculação mecânica foram feitos para tentar estabelecer infecção pela bactéria nas plantas de Bidens pilosa L., Parthenium hysterophorus L., Raphanus sativus L., Euphorbia heterophylla L., Sida rhombifolia L., Solanum americanum Mill. e Lantana camara L. Após meses da inoculação, as plantas foram testadas por PCR e isolamento primários para detectar a infecção por X. fastidiosa. Avaliou-se, também, a ocorrência de transmissão de X. fastidiosa de ameixeiras paras plantas infestantes, por cigarrinhas sabidamente vetoras, Sibovia sagata (Signoret) e Macugonalia cavifrons (Stål). Nas amostragens com rede de varredura, encontraram-se 72 espécies de cigarrinhas associadas às plantas infestantes dos pomares de ameixeiras, pertencentes às famílias Achilidae, Cercopidae, Cicadellidae, Delphacidae, Derbidae, Dictyopharidae, Flatidae e Membracidae. As cigarrinhas foram observadas em um total de oito espécies herbáceas de dicotiledôneas e sete monocotiledôneas. As plantas infestantes que abrigam maiores números de cigarrinha são Paspalum notatum Flügge, Parthenium hysterophorus L. e Raphanus sativus L. Dentre as espécies com potencial de transmitir X. fastidiosa, o cercopídeo Deois schach (Fabricius) e o cicadelíneo Plesiommata corniculata Young predominaram nas plantas infestantes dos pomares de ameixeira, podendo ter um papel chave em uma eventual disseminação primária de X. fastidiosa dessas plantas para ameixeira. Os cicadelíneos Acrogonia citrina Marucci & Cavichioli e Oncometopia facialis (Signoret) predominaram em capturas com armadilhas adesivas amarelas na copa das ameixeiras, o que sugere sua participação na disseminação secundária de X. fastidiosa entre árvores de ameixeira. O. facialis foi visualizada nos ramos de ameixeiras e de Lantana camara L. As plantas infestantes Solanum americanum Mill e L. camara permitem colonização por X. fastidiosa após inoculação mecânica. A cigarrinha Sibovia sagata (Signoret) é capaz de transmitir X. fastidiosa de ameixeira para S. americanum. / Plum Leaf Scald (PLS) is one of the major diseases that impair the production of plums in Brazil, caused by the xylem-limited bacterium Xylella fastidiosa (Wells), whose potential vectors in plums are sharpshooter leafhoppers (Hemiptera: Cicadellidae, Cicadellinae) and spittlebugs (Hemiptera: Cercopidae). A number of leafhoppers and and spittlebugs have been reported in Brazilian plum orchards, but their host plants and role in PLS epidemiology are largely unknown. The goal of this research was to investigate the association of potential hopper vectors with plum trees and weedy plants in the ground vegetation of orchards, in order to determine key vector species and weeds involved in PLS epidemiology. Therefore, a hopper survey was carried out in three plum orchards in the municipality of Paranapanema, SP, from September/2012 to April/2013, using three sampling methods: a) sweep net on weed species of the ground vegetation; b) yellow sticky cards placed on the plum canopy at 0.5 and 2 m above ground; and c) visual inspections of plum trees and some weeds. The collected hoppers were sorted and identified in the laboratory, and the data were submitted to faunistic analysis. To check if the weeds were hosts of plum strains of X. fastidiosa, bacterial suspensions were mechanically inoculated in Bidens pilosaL., Parthenium hysterophorus L., Raphanus sativus L., Euphorbia heterophylla L., Sida rhombifolia L., Solanum americanum Mill. Lantana camara L. The plants were assayed for infection of X. fastidiosa by PCR and culture at 2 months after inoculation. Transmission assays of X. fastidiosa from plum to weeds were carried out using two sharpshooter vectors, Sibovia sagata (Signoret) e Macugonalia cavifrons (Stål). The sweep net samplings revealed 72 species of seven hopper families (Achilidae, Cercopidae, Cicadellidae, Delphacidae, Derbidae, Dictyopharidae, Flatidae and Membracidae) associated with eight dicotyledoneous and seven monocotyledoneous weeds in the ground vegetation, with prevalence of Cicadellidae and Cercopidae. Among the potential hopper vectors, the spittlebug Deois schach (Fabricius) and the sharpshooter Plesiommata corniculata Young were predomimant species on the weed species, suggesting that they may play a key role in a possible primary spread of X. fastidiosa from weeds to plum. Paspalum notatum Flügge, Parthenium hysterophorus L. and Raphanus sativus L. were the weed species with the largest hopper populations. The sharpshooters Acrogonia citrina Marucci & Cavichioli e Oncometopia facialis (Signoret) were prevalent species trapped by the yellow sticky cards on the plum canopy, indicating that they may be involved in secondary spread of X. fastidiosa between plum trees in that region. O. facialis was visually detected on branches of plum trees and on the weed Lantana camara L. The weeds Solanum americanum Mill e L. camara allow colonization by X. fastidiosa after mechanical inoculation.The sharpshooter Sibovia sagata (Signoret) transmitted X. fastidiosa from plum to S. americanum.

A Escaldadura das Folhas da Ameixeira é o principal problema fitossanitário para a produção de ameixas no Brasil. A doença é causada pela bactéria Xylella fastidiosa, transmitida por cigarrinhas (Hemiptera: Cicadellidae, Cicadellinae). Neste trabalho foi realizado o levantamento, análise faunística e a flutuação populacional de Cicadellinae em pomares de ameixeira localizados nos Estados do Rio Grande do Sul (RS) e São Paulo (SP). A captura das cigarrinhas foi realizada com cartões adesivos amarelos (8,5 x 11,5 cm) em dois pomares de ameixeira de cada estado, localizados nos municípios de Paranapanema (SP) e Bento Gonçalves e Farroupilha (RS). Em cada pomar, foram instalados 20 cartões, distribuídos em 10 pontos espaçados 35 x 35 m, com duas alturas de amostragem (0,5 e 1,7m acima do nível do solo). Os cartões adesivos foram trocados quinzenalmente no período de setembro de 2006 a setembro de 2008. Com base na análise faunística, as espécies de cigarrinhas mais abundantes, constantes, freqüentes e dominantes foram analisadas para se conhecer a sua flutuação populacional. Nos dois pomares do RS foram coletados 1103 indivíduos de Cicadellinae distribuídos em 27 espécies. Destas, Erythrogonia dorsalis (Signoret, 1853), Sibovia sagata (Signoret,1854), Spinagonalia rubrovittata Cavichioli, 2008, Macugonalia cavifrons Stål, 1862, Dilobopterus dispar (Germar, 1821), Bucephalogonia xanthophis (Berg, 1879) e Molomea lineiceps Young, 1968 foram predominantes. Em SP, foram coletados 293 cicadelíneos, distribuídos em 10 espécies, sendo Oncometopia facialis (Signoret, 1854) e Molomea consolida Schröder, 1959 predominantes. Verificou-se uma menor diversidade e abundância de espécies de Cicadellinae nos pomares localizados em SP quando comparados com os pomares do RS. A distribuição populacional das espécies predominantes nos pomares do RS concentrou-se nos meses de janeiro a março, e em SP, de outubro a janeiro, correspondendo aos períodos de colheita e póscolheita em ambas as regiões. Os menores índices de captura de cicadelíneos foram obtidos nos meses de entressafra em todos os pomares amostrados. / The plum leaf scald (PLF) is a major threat for plum production in Brazil. PLF is caused by the bacteria Xylella fastidiosa which is transmitted by sharpshooter leafhoppers (Hemiptera: Cicadellidae, Cicadellinae). In this investigation, we surveyed the Cicadellinae species associated with plum orchards located in Rio Grande do Sul (RS) and São Paulo (SP) states, Brazil, in order to identify potential vectors of X. fastidiosa in these regions. We also studied the seasonal fluctuation of predominant species determined by faunistic analysis. In each area, adult sharpshooters were sampled using yellow sticky cards (8,5 x 11,5 cm) in two orchards in Paranapanema (SP) and one in Bento Gonçalves and Farroupilha (RS), respectivelly. In all orchards, traps were spaced 35 m apart in 10 sample units. Vertical distribution was also evaluated by placing cards at 0.5 and 1.7 m above soil level in each sample unit. The survey was conducted during two years from September 2006 to September 2008, by removing sticky cards fortnightly. In plum orchards located in RS state, we collected a total of 1103 specimens of 27 sharpshooter species. Erythrogonia dorsalis (Signoret, 1853), Sibovia sagata (Signoret,1854), Spinagonalia rubrovittata Cavichioli, 2008, Macugonalia cavifrons Stål, 1862, Dilobopterus dispar (Germar, 1821), Bucephalogonia xanthophis (Berg, 1879) and Molomea lineiceps Young, 1968 were predominant in these orchards. We collected 293 individuals of 10 sharpshooter species in SP state, where Oncometopia facialis (Signoret, 1854) and Molomea consolida Schröder, 1959 were predominant. We found a lower diversity and abundance of Cicadellinae species in plum orchards located in SP when compared to RS state. The predominant species were more abundant from January to March in RS and, from October to January in SP, corresponding to plum harvest and pos-harvest periods in both regions. The lowest population of sharpshooters in plum orchards was observed during the plant dormancy period.

Thesis (PhDAgric)--University of Stellenbosch, 2000. / ENGLISH ABSTRACT: Barley leaf scald, caused by Rhynchosporium secalis, is the most important disease of barley (Hordeum vulgare) in the Western Cape province of South Africa. The disease was first reported from South Africa in 1937. The present study is the first attempt to characterise the South African R. secalis population. Topics such as pathogenesisrelated proteins, virulence spectra, variability of pathotypes, sources of variation, host resistance, breeding strategies, molecular characterisation and fungicide sensitivity are summarised in Part 1 of this dissertation. In succeeding Parts the focus is on the characteristics of the local R. secalis population regarding virulence spectrum, DNA polymorphisms, in vitro as well as in vivo fungicide sensitivity. These aspects are treated as separate entities, leading to some duplication which is unavoidable. In Part 2 the virulence spectra of 50 R. secalis isolates from a population in the. Western Cape province were determined. Twenty-one races were detected using 17 differential barley cultivars. The two most prevalent races, namely races 4 and 7 had three and four virulence genes respectively. Both race 4 and 7 were virulent on the most susceptible cultivars, namely West China, Steudelli, C.I.8618 and C.I.2226. Considering the resistance genes reported for cultivars Atlas 46, Turk, and C.I.3515 which showed no susceptible cultivar-pathogen interaction, it would appear that the Rh- Rh3-Rh4 complex is primarily involved in conferring resistance to the local R. secalis isolates. A total of 20 races (47 isolates) characterised in Part 2 were selected for further characterisation by means of DNA fingerprinting. In Part 3 an anonymous multilocus DNA probe was used to characterise the genotypic structure of these isolates by means of RFLP analysis. No correlation between any particular fingerprint pattern, race, district, field or lesion was found. The two most prevalent races, 4 and 7, did not share the same genotypes, even when isolated from the same field or lesion. The genotypic diversity of the isolates studied was 46.5% of the theoretical maximum diversity. The high level of genotypic variation observed in the South African R. secalis population resembled the genotypic diversity observed in other cereal pathogens with known sexual structures. Although no teleomorph has yet been observed, these data suggest that sexual recombination may operate within the local population of R. secalis. In South Africa barley scald is primarily controlled by means of fungicides. The continued use of fungicides on cereal crops results in the build-up of fungicide resistance in the population, which could lower the efficacy of these compounds. These aspects were investigated in Part 4, where isolates (collected during 1993 to 1995) were evaluated in vitro for sensitivity to triadimenol, tebuconazole, flusilazole and propiconazole. The sensitivity fluctuated but in 1995 isolates were significantly less sensitive towards triadimenol than in the previous two years. In a second experiment, isolates collected from two fields with a 5-6 year-history of triadimenol seed treatments and tebuconazole applications, were evaluated for their fungicide sensitivity. A significant positive correlation was observed between tebuconazole and triadimenol sensitivity among,R. secalis populations from these fields. However, such a correlation was not found within the R. secalis population collected during 1993-1995 where shorter crop rotation patterns and a range of fungicides was applied. In a third experiment, the fungicide sensitivity of local R. secalis isolates was evaluated towards two new triazole fungicides, namely bromuconazole and triticonazole. Correlation coefficients observed between these new triazoles and those previously applied in South Africa were not significantly positive. The lack of significant cross-resistance has important practical implications regarding the management of fungicide resistance. In Part 5, isolates with different minimum inhibitory concentration (MIC) towards tebuconazole in vitro (1, 3 and 10 ug/ml) were compared in vivo. The aim of this study was to determine how MIC values would influence virulence (leaf area affected) and sporulation. Results indicated that all isolates were equally fit to induce lesions and sporulate in the absence of tebuconazole. Thus no fitness cost was associated with the degree of tebuconazole sensitivity in the present study. All R. secalis isolates were able to induce lesions on tebuconazole treated leaves, but differed significantly with respect to the percentage leaf area affected. Isolates, least sensitive (MIC = 10 ug/rnl) towards tebuconazole were more adapted on tebuconazole treated leaves, being able to repeatedly cause larger lesions than sensitive R. secalis isolates (MIC = 1 ug/rnl), Sporulation was not significantly different between isolates on lesions of untreated or tebuconazole treated leaves. Larger leaf areas affected and adequate sporulation suggest that a less sensitive population would result in more disease in tebuconazole treated fields. In conclusion, this study revealed the variability associated with the South African R. secalis population regarding virulence spectrum and genotypic structure. The data in this study suggest that it is likely that the local population will easily adapt to newly introduced, single gene resistance. For more durable resistance, higher levels of quantitative resistance should be introduced. This type of resistance is, however, more difficult to identify and incorporate than single gene resistance. Consequently, barley scald control will remain dependent on the efficacy of fungicide applications. Furthermore, the lack of cross-resistance and low frequency of resistant isolates indicates a low risk for the development of fungicide resistance in the local R. secalis population. Other factors such as current crop rotation practices and the range of fungicides being ~pplied also contribute to this low risk level. However, the status of these factors can change over time. The in vivo tebuconazole sensitivity study has indicated that a resistant field population of R. secalis may be able to build-up. It is, therefore, necessary to monitor the fungicide sensitivity of R. secalis isolates at timely intervals with view to successful barley cultivation in the future. / AFRIKAANSE OPSOMMING: Blaarvlek op gars (Hordeum vulgare), veroorsaak deur Rhynchosporium secalis, is die belangrikste siekte van gars in die Wes-Kaap provinsie van Suid-Afrika. Die voorkoms van R. secalis op gars is in Suid-Afrika vir die eertse keer in 1937 gerapporteer. Hierdie studie is die eerste poging tot karakterisering van die plaaslike R. secalis-populasie. Aspekte soos proteïene betrokke by patogenese, virulensiespektra, variabiliteit van patotipes, bronne van variasie, gasheerweerstand, teeltprogramme, molekulêre karakterisering en swamdodersensitiwiteit word in Deel I van die tesis opgesom. In die daaropvolgende gedeelte is die fokus op die karakterisering van die R. secalis-populasie en behels DNA karakterisering, virulensiespektrum, en swamdodersensitiwiteit in vitro asook in vivo. .. In Deel 2 is die virulensiespektra van 50 R. secalis isolate van 'n populasie in die. Wes-Kaap geëvalueer teenoor 17 differensiëel weerstandbiedende gars kultivars en hieruit is 21 rasse geïdentifiseer. Die twee mees algemene rasse (rasse 4 en 7), met onderskeidelik drie en vier virulensie gene, het virulent vertoon teenoor die mees vatbare kultivars soos West China, Steudelli, C.I.8618 en C.I.2226. Geen vatbare kultivar-patogeen interaksies is met kultivars Atlas 46, Turk en C.I.3515, wat al drie die Rh-Rh3-Rh4 kompleks dra, gevind nie. Dit wil dus voorkom asof hierdie genekompleks effektiewe gasheerweerstand teen die plaaslike R. secalis isolate kan bied. 'n Totaal van 20 rasse (47 isolate), gekarakteriseer in Deel 2, is geselekteer vir verdere karakterisering met behulp van DNA bandpatrone. In Deel 3 is 'n anonieme multilokus DNA peiler gebruik om deur middel van RFLP analise die genotipiese struktuur van hierdie R. secalis-isolate te bepaal. Geen assosiasie is gevind tussen DNA bandpatroon en ras, distrik, garsland of letsel nie. Die twee rasse (4 en 7) wat mees algemeen voorkom, het nie dieselfde bandpatroon vertoon nie, ook nie dié afkomstig vanuit dieselfde garsland of letsel nie. Die genotipiese diversiteit van isolate was 46.5% van die teoretiese maksimum diversiteit. Die hoë vlak van variasie waargeneem in die R. secalis populasie is soortgelyk aan variasie waargeneem in ander graanpatogene wat oor 'n geslagtelike stadium in die lewenssiklus beskik. Alhoewel geen geslagtelike stadium tot dusver geidentifiseer is nie, dui die vlak van variasie daarop dat geslagtelike rekombinasie moontlik wel plaasvind binne die plaaslike R. secalis populasie. In Suid-Afrika word blaarvlek op gars primêr deur swamdoders beheer. Die toenemende gebruik van swamdoders op graangewasse veroorsaak moontlik 'n opbou van swamdoderweerstand in die populasie. Dit kan die effektiwiteit van swamdoders verlaag. Hierdie veronderstelling is in Deel 4 ondersoek, waar die sensitiwiteit van isolate in vitro teenoor triadimenol, tebukonasool, flusilasool en propikonasool geëvalueer is. Die triasooi sensitiwiteit van R. secalis isolate wat gedurende die 1993- 1995 seisoen versamel is het gewissel en slegs vir triadimenol was daar 'n tendens na meer weerstandbiedenheid. 'n Swamdoder-evaluasie is in 'n aparte eksperiment op isolate gedoen wat versamel is vanaf twee garslande met 'n 5-6 jaar geskiedenis van triadimenol saadbehandelings en tebukonasool bespuitings. 'n Betekenisvolle positiewe korrelasie is waaJ~geneem tussen tebukonasool en triadimenol sensitiwiteit in R. secalis isolate afkomstig vanaf hierdie twee garslande. 'n Soortgelyke korrelasie is egter nie gevind in die populasie wat gedurende die 1993-1995 seisoene versamel IS me. Laasgenoemde kan moontlik toegeskryf word aan korter wisselboupatrone en die toediening van 'n verskeidenheid van swamdoders. In 'n derde eksperiment is die sensitiwiteit van plaaslike R. secalis isolate teenoor twee nuwe triasole, naamlik bromukonasool en tritikonasool getoets. Die korrelasie waargeneem tussen die twee nuwe triasole en triasooi swamdoders reeds voorheen in gebruik in die Wes-Kaap was me betekenisvol positief me. Die gebrek aan betekenisvolle kruisweerstandbiedendheid het belangrike praktiese implikasies vir die bestuur van swamdoder -weerstandbiedendheid. In Deel 5 is isolate met wisselende minimum inhiberende konsentrasies (MIKs) teenoor tebukonasool in vitro (1, 3 en 10 ug/ml) en in vivo vergelyk. Die doel van hierdie studie was om te bepaal hoe wisselende MIK-waardes virulensie (blaaroppervlakte geïnfekteer) en sporulasie sal beïnvloed. Resultate dui daarop dat alle R. secalis isolate in hierdie studie ewe fiks was om, in die afwesigheid van tebukonasool, letsels te induseer en te sporuleer. Die bevinding is dat die verlies in fiksheid nie geassosieer is met die mate van tebukonasool weerstand nie. Alle R. secalis isolate het die vermoë gehad om letsels op tebukonasool-behandelde blare te veroorsaak maar het betekenisvol verskil ten opsigte van die blaaroppervlakte geaffekteer. Isolate wat minder sensitief (MIK = 10 ug/rnl) teenoor tebukonasool in vitro is, het meer aangepastheid op tebukonasool-behandelde blare getoon. Gevolglik het hierdie isolate herhaaldelik meer letsels veroorsaak as sensitiewe isolate (MIK = 1 ug/ml), Sporulasie het nie betekenisvol verskil tussen isolate vanaf letsels op ondehandelde of tebukonsoolbehandelde blare nie. Hierdie resultate dui egter daarop dat 'n minder sensitiewe populasie tot meer siektevoorkoms in tebukonasool-bespuite lande kan lei. Die studie het die veranderlike karakter van die Suid-Afrikaanse R. secalispopulasie aangaande virulensiespektrum en genotipiese struktuur blootgelê. Dit is dus baie moontlik dat die R. secalis-populasie maklik sal aanpas by teelmateriaal met nuwe enkelgeen-weerstand. Vir volgehoue gasheerweerstand is dit egter nodig dat hoër vlakke van kwantitatiewe weerstand ingeteel moet word. In die praktyk is hierdie tipe weerstand egter baie moeiliker om te identifiseer en by nuwe teelmateriaal in te sluit as in die geval van enkelgeen-weerstand, Dit bring mee dat blaarvlekbeheer afhanklik bly van swamdodertoedienings as beheermaatreël. Die resultate van hierdie studie dui daarop dat daar tans 'n lae risiko vir die ontwikkeling van swamdoderweerstand in die plaaslike populasie is, as gevolg van die afwesigheid van kruisweerstandbiedendheid en die lae voorkoms van weerstandbiediende isolate. Ander faktore soos die wisselboustelsels wat toegepas word en die verskeidenheid van swamdoders toegedien dra ook daartoe by. Ten spyte hiervan kan die status van hierdie faktore egter oor tyd verander. Die in vivo tebukonasool studie het daarop gedui dat 'n weerstandbiedende veldpopulasie van R. secalis die potensiaal het om te vermeerder. Gevolglik is die tydige monitering van swamdodersenisitiwiteit van R. secalis isolate noodsaaklik om 'n volhoubare garsproduksie te verseker.

A escaldadura das folhas, causada por Xanthomonas albilineans, é uma importante doença da cana-de-açúcar, a qual interfere no rendimento e na longevidade de plantas infectadas. A intensidade de doença é variável, especialmente em relação a fatores climáticos e variedades cultivadas. A quantificação de danos é difícil de ser feita através de observações de plantas no campo. O uso de variedades resistentes tem sido eficiente para controle da doença, mas a diversidade do patógeno pode promover a quebra desta resistência. Em campos comerciais localizados no Estado de São Paulo, tem sido observado que a mesma variedade de cana é resistente à escaldadura em uma região, mas suscetível em outra. Este tipo de comportamento tem sugerido a ocorrência de variantes de X. albilineans que podem ser responsáveis pela quebra da resistência das variedades cultivadas. Seria relevante determinar a ocorrência de variabilidade para este patógeno dentro do estado de São Paulo, pois esta informação seria útil para indicação de variedades para as diferentes regiões e para orientar programas de melhoramento visando a obtenção de variedades resistentes. Com base nestas considerações, o objetivo do presente estudo foi investigar a diversidade genética de isolados coletados em áreas comerciais, usando técnicas moleculares como PCR, rep-PCR, RFLP e testes de patogenicidade. Os resultados mostraram que todos os isolados de plantas que exibiam sintomas de escaldadura eram pertencentes à espécie X. albilineans. Ainda, foi possível determinar a ocorrência de diversidade genética e variabilidade patogênica entre os isolados amostrados nas diferentes regiões. Os resultados obtidos no presente trabalho podem contribuir para novas investigações visando confirmar a associação da quebra da resistência de variedades com a variabilidade do patógeno. / Leaf scald, caused by Xanthomonas albilineans, is an important disease of sugarcane, That can interfere on the yield and longevity of infected plants. The disease intensity is variable, especially according to climate factors and cultivated varieties, and the damage quantification is very difficult through observation of plants in the field. Resistant varieties has been efficient to disease control, but the pathogen diversity can promote the breakdow of resistance. In commercial fields located in São Paulo State has been observed that the same sugarcane variety is resistant to leaf scald in a region, but susceptible in another one. This kind of behavior has suggested the occurrence of strains of X. albilineans that can be responsible for the breakdow of resistance of the cultivated varieties. It would be relevant to determinate pothogen variability in São Paulo State, because that information would be useful to indicate varieties for different regions and to guide breeding programs to obtain resistant varieties. Based upon the above points, the objective of the present study was to investigate the genetic diversity of bacterial isolates collected in commercial areas, using molecular techniques as PCR, rep-PCR, RFLP, and pathogenicity assays. The results showed that all isolates from plants exhibiting typical leaf scald symptoms belonged to species X. albilineans. It was also possible to determinate the occurrence of genetic diversity and pathogenic variability among the isolates sampled in the distinct regions. Thus, the results obtained in the present study can contribute to new investigations to confirm the association of the breakdow of resistance with the variability of the pathogen.

A Escaldadura das Folhas da Ameixeira (EFA) é a principal doença da cultura no Brasil, sendo causada pela bactéria Xylella fastidiosa e transmitida entre plantas pela ação de insetos vetores, mas há carência de informações sobre a identidade dos vetores e plantas hospedeiras para estirpes de X. fastidiosa causando EFA. Objetivando subsidiar uma proposta de manejo da EFA, foram realizados estudos sobre a transmissão de X. fastidiosa por vetores em ameixeira, identificação de plantas hospedeiras da bactéria em vegetação de cobertura dos pomares que possam servir como fontes de inóculo, capacidade de colonização de estirpes de ameixeira, cafeeiro e citros em inoculações cruzadas e validação da técnica de inoculação mecânica como método de avaliação de resistência de cultivars a X. fastidiosa em programas de melhoramento de ameixeira. Inicialmente, por meio de testes de colonização por X. fastidiosa foram identificadas Ocimum basilicum, Vernonia condensata e Pentas lanceolata como plantas não hospedeiras da bactéria, permitindo a criação de cigarrinhas sadias que foram utilizadas nos ensaios de transmissão. As cigarrinhas Macugonalia cavifrons, M. leucomelas e Sibovia sagata (Hemiptera: Cicadellidae: Cicadellinae) foram identificadas como vetoras de X. fastidiosa em ameixeira com eficiência de transmissão por indivíduo variando de 12 a 21%. Para identificação de hospedeiros alternativos do patógeno, avaliaram-se 12 espécies herbáceas (folhas e raízes) predominantes na vegetação de cobertura de pomares de ameixeira com elevada incidência de EFA, sendo dois no município de Videira, SC e três no município de Jarinú, SP. Amostras de nove delas (Bidens pilosa, Lepidium ruderale, Lolium multiflorum, Plantago major, Parthenium hysterosphorus, Raphanus sativus, Rumex sp., Solanum americanum e Vernonia sp. foram positivas para X. fastidiosa em folhas e/ou raízes, pelo teste de reação de polimerase em cadeia (PCR), Testes de inoculação mecânica com um isolado de X. fastidiosa de ameixeira comprovaram a capacidade dessa estirpe bacteriana em colonizar plantas de B. pilosa, L. ruderale, R. sativus e S. americanum. Estudos de inoculação cruzada demonstraram que isolados de X. fastidiosa de citros são capazes de colonizar cafeeiro, embora em menor proporção que isolados de cafeeiro. Um isolado de ameixeira foi capaz de colonizar citros e cafeeiro, sendo que em citros a proporção de plantas infectadas foi maior. Isolados de X. fastidiosa de citros e cafeeiro não colonizam plantas de ameixeira, sendo estas colonizadas apenas pelos isolados homólogos. Estudos de inoculação mecânica de dois isolados de X. fastidiosa de ameixeira em diferentes cultivares dessa planta hospedeira, expressaram resultados corroborativos com a pré-caracterização da resistência das cultivares, validando a técnica para uso em programas de melhoramento, com melhor precisão nos resultados e ganho de tempo na avaliação de resistência à EFA. / The Plum Leaf Scald (PLS) is the main disease of plum crop in Brazil being caused by the bacterium X. fastidiosa, which is transmitted among plants by insect vectors. However, additional information is needed about the identity of vectors and host plants of X. fastidiosa isolates that cause PLS. In order to subsidize PLS management proposal, studies on X. fastidiosa vector transmission in plum, identification of host plants from orchard vegetation cover that can be source of inoculum, bacterial colonization of strains isolated from plum, coffee and citrus in cross inoculations, and validation of mechanical inoculation technique as method for assessing resistance of plum varieties against X. fastidiosa in breeding programs, were carried out. Firstly, studies of colonization by X. fastidiosa revealed that three plants are non-hosts: Ocimum basilicum, Vernonia condensata and Pentas lanceolata, allowing the maintenance of healthy leafhoppers colonies. The sharpshooters Macugonalia cavifrons, M. leucomelas and Sibovia sagata were confirmed as vectors of X. fastidiosa in plum with transmission efficiency per individual ranging from 12 to 21%. In order to identify alternative hosts of the bacterium in plum orchards, 12 plant species (leaves and roots), which are predominant in the vegetation from orchards where PLS incidence is high, two of them being located at Videira - SC and three at Jarinu - SP, were chosen to be tested. Samples of leaves and/or roots from nine plant species (Bidens pilosa, Lepidium ruderale, Lolium multiflorum, Plantago major, Parthenium hysterosphorus, Raphanus sativus, Rumex sp., Solanum americanum and Vernonia sp.) were positive for X. fastidiosa by Polymerase Chain Reaction (PCR). Mechanical inoculation tests using a plum isolate of X. fastidiosa showed the capacity of this strain in colonizing the plants B. pilosa, L. ruderale, R. sativus and S. americanum. Studies on cross inoculation demonstrated that citrus X. fastidiosa isolates are able to colonize coffee plants, although in lower intensity than strains isolated from coffee. A plum isolate was able to colonize citrus and coffee plants, but the proportion of infected plants was higher in citrus. On the other hand, isolates of X. fastidiosa from citrus and coffee do not colonize plum plants and these were only colonized by homologous isolates. Studies on mechanical inoculation of two plum strains of X. fastidiosa in different plant cultivars revealed a pre-characterization on the resistance of these cultivars, validating the technique for use in breeding programs with a better precision in results as well as saving time on the assessment of resistance against PLS.

A Escaldadura das Folhas da Ameixeira (EFA) é a principal doença da cultura no Brasil, sendo causada pela bactéria Xylella fastidiosa e transmitida entre plantas pela ação de insetos vetores, mas há carência de informações sobre a identidade dos vetores e plantas hospedeiras para estirpes de X. fastidiosa causando EFA. Objetivando subsidiar uma proposta de manejo da EFA, foram realizados estudos sobre a transmissão de X. fastidiosa por vetores em ameixeira, identificação de plantas hospedeiras da bactéria em vegetação de cobertura dos pomares que possam servir como fontes de inóculo, capacidade de colonização de estirpes de ameixeira, cafeeiro e citros em inoculações cruzadas e validação da técnica de inoculação mecânica como método de avaliação de resistência de cultivars a X. fastidiosa em programas de melhoramento de ameixeira. Inicialmente, por meio de testes de colonização por X. fastidiosa foram identificadas Ocimum basilicum, Vernonia condensata e Pentas lanceolata como plantas não hospedeiras da bactéria, permitindo a criação de cigarrinhas sadias que foram utilizadas nos ensaios de transmissão. As cigarrinhas Macugonalia cavifrons, M. leucomelas e Sibovia sagata (Hemiptera: Cicadellidae: Cicadellinae) foram identificadas como vetoras de X. fastidiosa em ameixeira com eficiência de transmissão por indivíduo variando de 12 a 21%. Para identificação de hospedeiros alternativos do patógeno, avaliaram-se 12 espécies herbáceas (folhas e raízes) predominantes na vegetação de cobertura de pomares de ameixeira com elevada incidência de EFA, sendo dois no município de Videira, SC e três no município de Jarinú, SP. Amostras de nove delas (Bidens pilosa, Lepidium ruderale, Lolium multiflorum, Plantago major, Parthenium hysterosphorus, Raphanus sativus, Rumex sp., Solanum americanum e Vernonia sp. foram positivas para X. fastidiosa em folhas e/ou raízes, pelo teste de reação de polimerase em cadeia (PCR), Testes de inoculação mecânica com um isolado de X. fastidiosa de ameixeira comprovaram a capacidade dessa estirpe bacteriana em colonizar plantas de B. pilosa, L. ruderale, R. sativus e S. americanum. Estudos de inoculação cruzada demonstraram que isolados de X. fastidiosa de citros são capazes de colonizar cafeeiro, embora em menor proporção que isolados de cafeeiro. Um isolado de ameixeira foi capaz de colonizar citros e cafeeiro, sendo que em citros a proporção de plantas infectadas foi maior. Isolados de X. fastidiosa de citros e cafeeiro não colonizam plantas de ameixeira, sendo estas colonizadas apenas pelos isolados homólogos. Estudos de inoculação mecânica de dois isolados de X. fastidiosa de ameixeira em diferentes cultivares dessa planta hospedeira, expressaram resultados corroborativos com a pré-caracterização da resistência das cultivares, validando a técnica para uso em programas de melhoramento, com melhor precisão nos resultados e ganho de tempo na avaliação de resistência à EFA. / The Plum Leaf Scald (PLS) is the main disease of plum crop in Brazil being caused by the bacterium X. fastidiosa, which is transmitted among plants by insect vectors. However, additional information is needed about the identity of vectors and host plants of X. fastidiosa isolates that cause PLS. In order to subsidize PLS management proposal, studies on X. fastidiosa vector transmission in plum, identification of host plants from orchard vegetation cover that can be source of inoculum, bacterial colonization of strains isolated from plum, coffee and citrus in cross inoculations, and validation of mechanical inoculation technique as method for assessing resistance of plum varieties against X. fastidiosa in breeding programs, were carried out. Firstly, studies of colonization by X. fastidiosa revealed that three plants are non-hosts: Ocimum basilicum, Vernonia condensata and Pentas lanceolata, allowing the maintenance of healthy leafhoppers colonies. The sharpshooters Macugonalia cavifrons, M. leucomelas and Sibovia sagata were confirmed as vectors of X. fastidiosa in plum with transmission efficiency per individual ranging from 12 to 21%. In order to identify alternative hosts of the bacterium in plum orchards, 12 plant species (leaves and roots), which are predominant in the vegetation from orchards where PLS incidence is high, two of them being located at Videira - SC and three at Jarinu - SP, were chosen to be tested. Samples of leaves and/or roots from nine plant species (Bidens pilosa, Lepidium ruderale, Lolium multiflorum, Plantago major, Parthenium hysterosphorus, Raphanus sativus, Rumex sp., Solanum americanum and Vernonia sp.) were positive for X. fastidiosa by Polymerase Chain Reaction (PCR). Mechanical inoculation tests using a plum isolate of X. fastidiosa showed the capacity of this strain in colonizing the plants B. pilosa, L. ruderale, R. sativus and S. americanum. Studies on cross inoculation demonstrated that citrus X. fastidiosa isolates are able to colonize coffee plants, although in lower intensity than strains isolated from coffee. A plum isolate was able to colonize citrus and coffee plants, but the proportion of infected plants was higher in citrus. On the other hand, isolates of X. fastidiosa from citrus and coffee do not colonize plum plants and these were only colonized by homologous isolates. Studies on mechanical inoculation of two plum strains of X. fastidiosa in different plant cultivars revealed a pre-characterization on the resistance of these cultivars, validating the technique for use in breeding programs with a better precision in results as well as saving time on the assessment of resistance against PLS.

A escaldadura das folhas, causada por Xanthomonas albilineans, é uma importante doença da cana-de-açúcar, a qual interfere no rendimento e na longevidade de plantas infectadas. A intensidade de doença é variável, especialmente em relação a fatores climáticos e variedades cultivadas. A quantificação de danos é difícil de ser feita através de observações de plantas no campo. O uso de variedades resistentes tem sido eficiente para controle da doença, mas a diversidade do patógeno pode promover a quebra desta resistência. Em campos comerciais localizados no Estado de São Paulo, tem sido observado que a mesma variedade de cana é resistente à escaldadura em uma região, mas suscetível em outra. Este tipo de comportamento tem sugerido a ocorrência de variantes de X. albilineans que podem ser responsáveis pela quebra da resistência das variedades cultivadas. Seria relevante determinar a ocorrência de variabilidade para este patógeno dentro do estado de São Paulo, pois esta informação seria útil para indicação de variedades para as diferentes regiões e para orientar programas de melhoramento visando a obtenção de variedades resistentes. Com base nestas considerações, o objetivo do presente estudo foi investigar a diversidade genética de isolados coletados em áreas comerciais, usando técnicas moleculares como PCR, rep-PCR, RFLP e testes de patogenicidade. Os resultados mostraram que todos os isolados de plantas que exibiam sintomas de escaldadura eram pertencentes à espécie X. albilineans. Ainda, foi possível determinar a ocorrência de diversidade genética e variabilidade patogênica entre os isolados amostrados nas diferentes regiões. Os resultados obtidos no presente trabalho podem contribuir para novas investigações visando confirmar a associação da quebra da resistência de variedades com a variabilidade do patógeno. / Leaf scald, caused by Xanthomonas albilineans, is an important disease of sugarcane, That can interfere on the yield and longevity of infected plants. The disease intensity is variable, especially according to climate factors and cultivated varieties, and the damage quantification is very difficult through observation of plants in the field. Resistant varieties has been efficient to disease control, but the pathogen diversity can promote the breakdow of resistance. In commercial fields located in São Paulo State has been observed that the same sugarcane variety is resistant to leaf scald in a region, but susceptible in another one. This kind of behavior has suggested the occurrence of strains of X. albilineans that can be responsible for the breakdow of resistance of the cultivated varieties. It would be relevant to determinate pothogen variability in São Paulo State, because that information would be useful to indicate varieties for different regions and to guide breeding programs to obtain resistant varieties. Based upon the above points, the objective of the present study was to investigate the genetic diversity of bacterial isolates collected in commercial areas, using molecular techniques as PCR, rep-PCR, RFLP, and pathogenicity assays. The results showed that all isolates from plants exhibiting typical leaf scald symptoms belonged to species X. albilineans. It was also possible to determinate the occurrence of genetic diversity and pathogenic variability among the isolates sampled in the distinct regions. Thus, the results obtained in the present study can contribute to new investigations to confirm the association of the breakdow of resistance with the variability of the pathogen.

Xanthomonas albilineans est la bactérie responsable de l'échaudure des feuilles, une maladie vasculaire et létale de la canne à sucre. Cette plante, domestiquée depuis plus de 5000 ans, est aujourd'hui cultivée sur près de 26 millions d'hectares. Depuis le début du 20ème siècle, les variétés naturelles de canne à sucre ont été remplacées par des variétés hybrides issues de croisements artificiels et présentant des caractères agronomiques plus intéressants. La transmission de X. albilineans est essentiellement liée au mode de multiplication par bouturage de la canne à sucre, les boutures produites à partir d'un plant infecté ayant de fortes chances d'être contaminées. L'utilisation d'outils de coupe favorise également la transmission de la bactérie à partir d'un plant infecté. Une transmission aérienne a toutefois été décrite pour au moins un groupe génétique de souches de X. albilineans appelé PFGE-B et caractérisé à l'aide de la technique d'électrophorèse en champ pulsé. Mon sujet de thèse visait, par une étude de génomique comparative et évolutive sur plusieurs souches de X. albilineans, à, d'une part, comprendre l'histoire de l'association de cette bactérie avec la canne à sucre, et, d'autre part, identifier des gènes candidats qui pourraient expliquer la pathogénie de X. albilineans et la capacité des souches du groupe PFGE-B à être transmises par voie aérienne.En comparant 12 souches de X. albilineans appartenant à neuf groupes PFGE différents, j’ai construit une phylogénie solide proposant l'existence de huit lignées regroupées en deux clades. Cette phylogénie a été confirmée en utilisant trois méthodes de phylogénomique différentes. De façon intéressante, cette phylogénie permet de proposer des hypothèses sur l'évolution de X. albilineans et sur l'introduction de cette bactérie chez la canne à sucre. L'analyse des gènes présents dans le génome de ces 12 souches appartenant à neuf groupes PFGE différents m'a également permis d'identifier un système RM (restriction/modification) qui n'est présent que chez les souches PFGE-B et qui pourrait expliquer pourquoi ces souches sont plus résistantes aux phages que celles appartenant aux autres groupes PFGE. En facilitant la survie des bactéries dans l'environnement, cette résistance aux phages pourrait jouer un rôle important dans la transmission aérienne.La comparaison de 11 souches de X. albilineans appartenant au groupe PFGE-B m'a permis de construire, à partir de 67 SNP (Single Nucleotide Polymorphism), une phylogénie qui indique que les Caraïbes, où la canne à sucre n'a été introduite qu'à la fin du 15ème siècle, pourraient être un centre de dispersion des souches PFGE-B. Cette étude a également mis en évidence un polymorphisme au niveau des spacers d'un locus CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats system). L'étude de ce polymorphisme sur 18 souches de X. albilineans non séquencées a permis de démontrer que les souches de Floride sont originaires des Caraïbes et de proposer des hypothèses concernant l'origine des souches PFGE-B d’autres régions du monde, même si le séquençage d'un plus grand nombre de souches sera nécessaire pour confirmer ces hypothèses.En analysant la séquence du génome de deux souches qui avaient été initialement classées dans l'espèce X. albilineans mais qui n'avaient pas été isolées à partir de tiges de canne à sucre, j’ai mis en évidence une nouvelle espèce que nous avons appelée 'Xanthomonas pseudalbilineans'. Enfin, la comparaison du génome fini et annoté de la souche PFGE-B GPE PC73 avec le génome d'autres espèces bactériennes phytopathogènes m'a permis de proposer des hypothèses pour expliquer la pathogénie de X. albilineans. / Xanthomonas albilineans is the bacterium causing leaf scald, a vascular disease being lethal for sugarcane. This plant, domesticated for over 5,000 years, is nowadays grown across almost 26 million hectares. Since the beginning of 20th century, natural varieties of sugarcane have been replaced by hybrid varieties resulting from artificial crossings. The transmission of X. albilineans is mainly linked to the fact that sugarcane is propagated by taking cuttings, with the cuttings from an infected plant likely to be contaminated. The use of cutting instruments also facilitates transmission of the bacteria from an infected plant. An aerial transmission has nevertheless been described, at least for one genetic group of strains from X. albilineans, namely PFGE-B, characterized using the pulsed-field gel electrophoresis technique. My thesis topic consisted in performing a comparative and evolutive genomic study of several strains of X. albilineans in order, on one hand, to better understand the history of association of this bacterium with the sugarcane, and, on the other hand, to identify candidate genes which may explain the pathogenicity of X. albilineans as well as the ability of PFGE-B strains to be aerially transmitted. Comparing 12 strains of X. albilineans belonging to nine different PFGE groups, I built a robust phylogeny which proposes the existence of eight lineages clustered in two clades. This phylogeny has been confirmed by three different phylogenomic methods. Interestingly, this phylogeny allows to propose hypotheses about the evolution of X. albilineans and the introduction of this bacterium in sugarcane. The analysis of genes from the genomes of these 12 strains belonging to nine different PFGE groups also allowed me to identify a RM (Restriction/Modification) system which is present only in PFGE-B strains and which may explain why these strains are more resistant to phages that those belonging to other PFGE groups. By facilitating the survival of bacteria in the environment, this resistance to phages may play an important role in aerial transmission. The comparison of 11 strains of X. albilineans belonging to the PFGE-B group allowed me to build, based on 67 SNPs (Single Nucleotide Polymorphism), a phylogeny which indicates that the Caribbean area, where sugarcane was introduced not before the end of the 15th century, might be a center for dispersion of PFGE-B strains. This study also revealed a polymorphism of spacers of a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats system) locus. The study of this polymorphism in 18 non-sequenced strains of X. albilineans allowed to demonstrate that strains from Florida originated in the Caribbean region, and to propose hypotheses regarding the origin of PFGE-B strains from other regions of the world, even if the sequencing of a higher number of strains will be required to confirm these hypotheses.Analyzing the sequence of the genome of two strains, initially thought to belong to the X. albilineans species but which weren’t isolated from sugarcane stalks, I revealed a new species that we called ‘Xanthomonas pseudalbilineans’. Finally, the comparison of the finished and annotated genome of the PFGE-B strain GPE PC73 with the genome of other phytopathogenic bacteria allowed me to propose hypotheses to explain X. albilineans’ pathogenicity.