Dissertationen zum Thema „Black rot disease“

A quantitative technique has been developed to assay cotton soils for populations of Thielaviopsis basicola, a soil occurring fungus that causes the seedling disease of cotton known as Black Root Rot. The procedure utilizes a soil dilution technique with a carrot extract agar containing etridiazol, Mystatin, streptomycin sulfate, chlortetracycline, calcium carbonate and PCNB. Populations of the fungus have been monitored from April to December, 1986 in a heavily infested Pima S-6 field in cooperation with Bob Cockrill, a Coolidge grower. When field soils containing approximately 600 propagules of the fungus per gram of air dry soil were planted to Pima S-6 in the laboratory, 75-100% and 50-75% cortical decay occurred at 20 and 28 C, respectively. This seedling damage was related to subsequent reduced seedling vigor.

The 1985 and 1986 Cotton Reports have the same publication and P-Series numbers.
NU-Zone (imazalil) + Nu-Flow ND (TCMTB + Chloroneb), NU-Flow ND, and Vitavax (carboxin) were evaluated as seed treatments with and without in-furrow PCNB. The following was learned: - Vitavax-treated seed got out of the ground faster than the other treatments, which brings out the possibility that NU-Flow or NU-Zone slows germination. - Stands and root development were slightly better when NUZone was present. - NU-Zone + NU-Flow ND seed treatment is not totally effective in controlling black root rot in heavily inoculated soils. - NU-Flow ND alone is the least effective of the treatments. - In-furrow PCNB did not affect yields.

Two planting dates, March 28, and April 28 were used to study the effect of soil temperature during planting on black root rot of cotton. Also, several cotton varieties were evaluated for response to the disease under varying soil temperatures and inoculum levels.

L’ascomicete Guignardia bidwellii, agente causale del marciume nero della vite, è un patogeno economicamente importante in alcuni areali viticoli. La conoscenza, disponibile sul marciume nero dell’uva, è stata recuperata dalla letteratura, analizzata e sintetizzata per sviluppare un modello meccanicistico del ciclo di vita del patogeno, guidata dalle variabili meteorologiche e dalla fenologia della vite, e basata sull'analisi dei sistemi. Il modello è stato poi valutato per la sua capacità di rappresentare il sistema reale e la sua utilità per la comprensione di epidemie di marciume nero su foglie e grappoli in un vigneto del Nord Italia, nel 2013 al 2015. Successivamente, le mancanze di conoscenza sono state analizzate, studiate e quindi colmate attraverso specifici esperimenti. In un primo passo, le dinamiche dell’inoculo primario e dei modelli di dispersione (di entrambi ascospore e conidi) da mummie svernate sono state studiate in un vigneto sperimentale per tre anni. In un secondo passo, l'effetto della temperatura e dell'umidità sulla formazione di picnidi di G. bidwellii e la successiva estrusione di cirri, nelle lesioni su foglia, la produzione e la germinazione dei conidi (inoculo secondario), e la lunghezza del periodo di latenza sono stati studiati sia in condizioni di campo che in ambiente controllato. In un terzo passo, sono stati condotti studi in ambiente controllato per studiare la produzione di conidi di G. bidwellii sulle lesioni di foglie, influenzata da lavaggi ripetuti e alternando periodi di secco ed umido. Il modello epidemiologico sviluppato in questa tesi può essere utilizzata da viticoltori come strumento predittivo per la pianificazione di trattamenti fungicidi nei vigneti.
The ascomycete Guignardia bidwellii, causal agent of black-rot on grapevines, is an economically important pathogen in some viticultural areas. The available knowledge on black-rot of grape was retrieved from literature, analyzed, and synthesized to develop a mechanistic model of the life cycle of the pathogen, driven by weather and vine phenology, and based on the systems analysis. The model was then evaluated for its ability to represent the real system and its usefulness for understanding black-rot epidemics on leaves and bunches in a vineyard of north Italy, in 2013 to 2015. Thereafter, weaknesses in our knowledge were analysed and studied through specific experiments. In a first step, dynamics of primary inoculum and dispersal patterns (both ascospores and conidia) from overwintered grape mummies were investigated in an experimental vineyard during three years. In a second step, the effect of temperature and humidity on the formation of G. bidwellii pycnidia and the extrusion of cirri in grape leaf lesions, production and germination of conidia (secondary inoculum), and the length of the latency period were studied under both environmental and controlled conditions. In a third step, environmental-controlled studies were conducted to investigate the production course of G. bidwellii conidia on grape leaf lesions as influenced by repeated washing events and alternate dry and wet periods. The model developed in this thesis can be used by vinegrowers as a predictive tool for scheduling fungicide sprays in the vineyards.

L’ascomicete Guignardia bidwellii, agente causale del marciume nero della vite, è un patogeno economicamente importante in alcuni areali viticoli. La conoscenza, disponibile sul marciume nero dell’uva, è stata recuperata dalla letteratura, analizzata e sintetizzata per sviluppare un modello meccanicistico del ciclo di vita del patogeno, guidata dalle variabili meteorologiche e dalla fenologia della vite, e basata sull'analisi dei sistemi. Il modello è stato poi valutato per la sua capacità di rappresentare il sistema reale e la sua utilità per la comprensione di epidemie di marciume nero su foglie e grappoli in un vigneto del Nord Italia, nel 2013 al 2015. Successivamente, le mancanze di conoscenza sono state analizzate, studiate e quindi colmate attraverso specifici esperimenti. In un primo passo, le dinamiche dell’inoculo primario e dei modelli di dispersione (di entrambi ascospore e conidi) da mummie svernate sono state studiate in un vigneto sperimentale per tre anni. In un secondo passo, l'effetto della temperatura e dell'umidità sulla formazione di picnidi di G. bidwellii e la successiva estrusione di cirri, nelle lesioni su foglia, la produzione e la germinazione dei conidi (inoculo secondario), e la lunghezza del periodo di latenza sono stati studiati sia in condizioni di campo che in ambiente controllato. In un terzo passo, sono stati condotti studi in ambiente controllato per studiare la produzione di conidi di G. bidwellii sulle lesioni di foglie, influenzata da lavaggi ripetuti e alternando periodi di secco ed umido. Il modello epidemiologico sviluppato in questa tesi può essere utilizzata da viticoltori come strumento predittivo per la pianificazione di trattamenti fungicidi nei vigneti.
The ascomycete Guignardia bidwellii, causal agent of black-rot on grapevines, is an economically important pathogen in some viticultural areas. The available knowledge on black-rot of grape was retrieved from literature, analyzed, and synthesized to develop a mechanistic model of the life cycle of the pathogen, driven by weather and vine phenology, and based on the systems analysis. The model was then evaluated for its ability to represent the real system and its usefulness for understanding black-rot epidemics on leaves and bunches in a vineyard of north Italy, in 2013 to 2015. Thereafter, weaknesses in our knowledge were analysed and studied through specific experiments. In a first step, dynamics of primary inoculum and dispersal patterns (both ascospores and conidia) from overwintered grape mummies were investigated in an experimental vineyard during three years. In a second step, the effect of temperature and humidity on the formation of G. bidwellii pycnidia and the extrusion of cirri in grape leaf lesions, production and germination of conidia (secondary inoculum), and the length of the latency period were studied under both environmental and controlled conditions. In a third step, environmental-controlled studies were conducted to investigate the production course of G. bidwellii conidia on grape leaf lesions as influenced by repeated washing events and alternate dry and wet periods. The model developed in this thesis can be used by vinegrowers as a predictive tool for scheduling fungicide sprays in the vineyards.

Thesis (MScAgric)--University of Stellenbosch, 2002.
ENGLISH ABSTRACT: Historically root diseases have been a production-limiting problem for the strawberry industry worldwide. In the Western Cape Province of South Africa the most serious root disease is black root rot, which causes losses of up to 30%, annually. The aims of this study were to investigate aspects of the etiology and epidemiology of this disease in the Western Cape, and to provide information that can be incorporated in an integrated disease management strategy. In Chapter I a summary of published information on this disease is presented. Disease symptoms include severe stunting of plants, which have black, rotted, reduced root systems. Even though this disease is of great economic importance, the etiology remains unresolved. However, soilborne fungal root pathogens, particularly Pythium and Rhizoctonia spp. have been implicated as major role players. Control of this disease, as well as the other root diseases affecting strawberries, has relied on soil fumigation with broad spectrum chemical fumigants, in particular methyl bromide. However, due to the ozone depleting action of methyl bromide it was decided at the signing of the Montreal Protocol to remove this chemical from the market. This action has caused great demand for alternative measures to control root diseases on many crops including strawberries. Development of integrated disease management strategies is dependent upon a more complete understanding of the etiology, biology and ecology of the disease complex. In Chapter 2 fungal pathogens associated with diseased plants were isolated and Koch's postulates were carried out. The most frequently isolated fungal pathogens were Pythium irregulare, Rhizoctonia spp. and Cylindrocarpon destructans. Two morphotypes of Rhizoctonia were isolated viz. a brown and a white type. Pythium irregulare was isolated more frequently in July than in September, and was not isolated at all in November. Rhizoctonia spp. were present at all sampling dates but were more frequently isolated in November than at the other times. All the fungi that were tested were pathogenic and caused root lesions. Cylindrocarpon destructans and Coniella fragariae did not have a stunting effect on the plants. These results confirm a major role for Pythium and Rhizoctonia in the black root rot complex and suggest that there is a complimentary seasonal variation in occurrence between these two pathogens. The Rhizoctonia species and anastomosis groups of isolates obtained from diseased strawberries in the Western Cape Province were determined, and their pathogenicity and relative virulence assessed. Both binucleate and multinucleate types were recovered from diseased roots and identified as R. fragariae and R. so/ani, respectively. All isolates of R. solani were members of anastomosis group (AG) 6, whereas three AG types were identified among isolates of R. fragariae, viz. AG-A, AG-G and AG-I at a relative occurrence of 69%, 25%, 6% respectively. All Rhizoctonia isolates were pathogenic to strawberry, but R. solani (AG 6) was the most virulent causing severe stunting of plants. This is the first species confirmation and AG type identification of Rhizoctonia taxa causing root rot of strawberries in South Africa. An assessment of the presence and quantity of black root rot pathogens associated with soils prior to fumigation and post fumigation with methyl bromide, was made in Chapter 4. Isolations were also made from nursery plants to determine whether any black root rot pathogens were in the plants before transplanting. Results demonstrated that after fumigation the soil was free of all pathogenic fungi associated with the disease. However, the main pathogens involved in black root rot, viz. Rhizoctonia fragariae, R. solani and Pythium spp. were isolated from nursery plants. The fact that the plants are already infected with these pathogens renders the prospects for control of this disease difficult. Further studies are urgently required to develop production practices that can be included in disease management programmes. In vitro studies were carried out to determine the ECso values of different fungicides for isolates of Pythium irregulare, Rhizoctonia fragariae AG-A, AG-G and AG-I and R. solani AG 6. Benomyl, fludioxonil and tolc1ofos-methyl were used in these tests. Field trials were also conducted using these fungicides. In Chapter 5 it is shown that in general application of fungicides improved the yield and did not affect the survival rate of strawberry plants. Fludioxonil showed potential for short-term use. Applications of fungicides that targeted specific fungal genera were not sufficient to control the disease. Seasonal fluctuation of Pythium and Rhizoctonia spp. became apparent with the occurrence of Pythium being relatively high early in the season but low late in the season. Conversely, the occurrence of Rhizoctonia was low at the beginning of the season but high late in the season. In the short-term there is potential for fungicide applications as part of an integrated disease management strategy, but the economic feasibility of this practice needs to be assessed. In this study the major pathogens causing black root rot were identified in the Western Cape Province of South Africa, and important information regarding the epidemiology of the disease was reported. These results can be incorporated in an integrated management strategy to reduce losses of strawberry production, attJibutable to black root rot.
AFRIKAANSE OPSOMMING: Wortelsiektes is wêreldwyd 'n produksie-beperkende probleem vir die aarbeibedryf. . Swartwortelvrot, wat jaarliks verliese van tot 30% veroorsaak, is die belangrikste wortelsiekte in die Wes-Kaap Provinsie van Suid-Afrika. Die doelwitte van hierdie studie was om aspekte van die etiologie en epidemiologie van die siekte in die Wes- Kaap te ondersoek en inligting wat in geïntegreerde siektebestuurstrategië ingesluit kan word, te voorsien. In Hoofstuk 1 word 'n opsomming van gepubliseerde inligting aangaande die siekte uiteengesit. Siektesimptome sluit ernstige verdwerging van plante met swart verotte en verkleinde wortelstelsels in. Alhoewel die siekte van groot ekonomiese belang is, is die etiologie grootliks onbekend. Grondgedraagde wortelpatogene swamme, spesifiek Pythium en Rhizoctonia spp., is egter as belangrike rolspelers geïdentifiseer. Tot dusver het die beheer van hierdie siekte sowel as ander wortelsiektes van aarbeie berus op grondberoking met breë spektrum chemiese berokingsmiddels, spesifiek metielbromied. As gevolg van die osoonafbrekende aksie van metielbromied is daar egter tydens die ondertekening van die Montreal Protocol besluit om dié middel van die mark te verwyder. Hierdie besluit het 'n groot aanvraag na alternatiewe beheermaatreëls vir wortelsiektes van verskeie gewasse, insluitende aarbeie, veroorsaak. Die ontwikkeling van geïntegreerd siektebestuurstrategieë is egter afhanklik van 'n meer volledige begrip van die etiologie, biologie en ekologie van die siektekompleks. In Hoofstuk 2 is die patogene swamme wat met die siekte geassosieer word, geïsoleer, en is Koch se postulate uitgevoer. Die mees algemeen geïsoleerde patogene swamme was Pythium irregulare, Rhizoctonia spp. en Cylindrocarpon destructans. Twee morfotipes van Rhizoctonia is geïsoleer, nl. 'n bruin tipe en 'n wit tipe. Pythium irregulare is meer dikwels in Julie as in September geïsoleer, maar glad nie in November nie. Rhizoetonia het tydens alle monstertye voorgekom, maar is meer dikwels in November geïsoleer. Al die swamme wat getoets is, was patogenies en het letsels op die wortels veroorsaak. Cylindroearpon des true tans en Coniella fragariae het nie'n verdwergingseffek op plante gehad nie. Hierdie resultate bevestig die dominante rol van Pythium en Rhizoctonia in die swartwortelvrot kompleks en dui op 'n komplementêre seisoenale variasie in die voorkoms van hierdie twee patogene. Die Rhizoctonia spesies en anastomose groepe (AG) van die isolate geisoleer vanaf siek aarbeiplante in die Wes-Kaap Provinsie is bepaal, en die patogenisiteit en relatiewe virulensie is beraam. Sowel tweekernige as multikernige tipes is vanaf siek wortels geïsoleer en respektiewelik as R. fragariae en R. so/ani geïdentifiseer. Alle isolate van R. so/ani was lede van anastomose groep 6, terwyl drie AG tipes, nl. AGA, AG-G en AG-I onder die R. fragariae isolate geïdentifiseer is met relatiewe voorkomste van 69%, 25%, 6% respektiewelik. Alle Rhizoctonia isolate was patogenies op aarbeie, maar R. so/ani (AG 6) was die mees virulente en het ernstige verdwerging van plante veroorsaak. Hierdie is die eerste bevestiging van spesies en identifisering van AG tipes van Rhizoctonia taksa wat wortelvrot van aarbeie in Suid Afrika veroorsaak. In Hoofstuk 4 is 'n beraming van die voorkoms en hoeveelheid swartwortelvrot patogene geassosieer met grond voor, en na beroking met metielbromied, gemaak. Isolasies is ook vanaf kwekeryplante gemaak om te bepaal of enige swartwortelvrot patogene voor oorplanting in die plante teenwoordig was. Die resultate het getoon dat grond na beroking vry was van alle patogeniese swamme geassosieër met die siekte. Die hoof patogene betrokke in die swartwortelvrot kompleks, nl. Rhizoctonia fragariae, R. so/ani en Pythium spp. was egter in die kwekery plante teenwoordig. Die feit dat plante reeds met hierdie patogene geïnfekteer is, maak die vooruitsigte vir die beheer van hierdie siekte moeilik. Verdere studies word dringend benodig vir die ontwikkeling van produksiepraktyke wat by siektebestuursprogramme ingesluit kan word. In vitro studies om die ECso waardes van die isolate van Pythium irregulare, Rhizoctonia fragariae AG-A, AG-G en AG-I en R. so/ani AG 6 vir die fungisiedes benomyl, fludioxonil en tolclofos-metiel te bepaal, is uitgevoer. Hierdie fungisiedes is ook in veldproewe getoets. In Hoofstuk 5 is getoon dat aanwending van fungisiedes die opbrengs verbeter het en nie die oorlewing van aarbeiplante beïnvloed het nie. Fludioxonil het potensiaal vir korttermyn gebruik getoon. Die aanwending van fungisiedes wat spesifieke swamgenera teiken, was nie voldoende om die siekte te beheer nie. Seisoenale fluktuasies van Pythium en Rhizoctonia spp. het duidelik geword met die relatief hoë voorkoms van Pythium vroeg in die seisoen, maar lae voorkoms laat in die seisoen, terwyl die voorkoms van Rhizoctonia laag was aan die begin van die seisoen, maar hoog later in die seisoen. In die korttermyn is daar potensiaal vir fungisiedtoedienings as deel van 'n geïntegreerde siektebestuurstrategie, maar die ekonomiese haalbaarheid van hierdie praktyk moet bepaal word. In hierdie studie is die hoof patogene wat swartwortelvrot van aarbeie in die Wes-Kaap Provinsie van Suid-Afrika veroorsaak geïdentifiseer, en belangrike inligting rakende die epidemiologie van die siekte is aangeteken. Hierdie resultate kan in 'n geïntegreerde bestuurstrategie geïnkorporeer word om verliese van aarbeiproduksie, toeskryfbaar aan swartwortelvrot te, verminder.

Incidence of Black Root Rot of cotton and soil inoculum potential of the causal agent Thielaviopsis basicola were monitored throughout the season in two adjacent fields planted in mid April, 1987 in Duncan, Arizona. Mean inoculum potential in Field 1 soil was 65 cfu/g and 20 % of the seedlings were infected. In Field 2 the inoculum potential and percentage of infected plants were 225 cfu/g and 93, respectively. No cortical decay was noted after June 6 in either field. Yields were similar in both fields.

A new selective medium (TB-CEN) was developed for isolating Thielaviopsis basicola, cause of black root rot of tobacco, from soil. TB-CEN medium contains etridiazol and nystatin to inhibit the growth of undesired fungi, and unautoclaved extract from carrot to selectively enhance for the growth of T. basicola. Inoculum and/or population densities of T. basicola in five burley tobacco fields were 74-166 propagules per g of soil, and 0-12 propagules per g of soil in three other burley fields. Inoculum and/or population densities of T. basicola in 12 flue-cured and 2 sun-cured tobacco fields were 0-26 propagules per g of soil, and 101 and 402 propagules per g of soil in two other flue-cured fields. Environmental factors apparently had a strong effect on black root rot development, since root rot and plant stunting were severe in two burley fields that had 148 and 158 propagules per g of soil, but were not severe in the two flue-cured fields that had 101 and 402 propagules per g of soil. All of the cultivars planted in the four fields were susceptible. Black root rot was the major disease associated with the stunting of tobacco plants in the burley region of Virginia, but not in the flue- and sun-cured regions. No evidence was found to indicate that endomycorrhizae were involved in tobacco stunting in Virginia. T basicola inoculum density-disease severity studies were conducted both in soil-temperature tanks and in the field. Tobacco seedlings were grown in temperature tanks (20-23 C) for 30-31 days in naturally infested field soil (pH 6.5). For all cultivars tested (Burley 21, NC95, and Va Gold), the mean percent of roots that were rotted increased significantly (P=0.001) as inoculum density increased (R² range for regressions=0.93-0.97). Severe levels of root rot occurred at inoculum densities of 50-200 propagules per g of soil. Significant (P=0.01) reductions in plant growth occurred at inoculum densities as low as 5-10 propagules per g of soil. In a study conducted on a commercial burley tobacco (cv. B21-Ky10) field, inoculum densities of 150 and 683 propagules per g of soil were associated with moderate and severe levels of black root rot, respectively. Differences between soil-temperature tank and field studies appeared to be due to variations in environmental- and host-related factors. In another burley field study, the fungicide imazalil, which completely inhibited the growth of T. basicola when amended into agar media at a concentration of 1.0 μg a.i./ml, failed to control black root rot when it was added to transplanting water (50 ml/plant) at concentrations as high as 1,500 μg a.i./ml.
Ph. D.
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High population levels of Rhizoctonia solani in a field in Coolidge, AZ which also contained Thielaviopsis basicola failed to cause the expected infection by R. solani in untreated seeds of Gossypium hirsutum. Growth chamber experiments with autoclaved soil amended with inoculum of both R. solani, and T. basicola had a significantly lower infection rate by R. solani than did the control soil with R. solani alone.

Neem cake is a product of the cold pressing from the neem kernels to obtain neem oil. Bio-active substances from neem cake extracted solutions were evaluated for their potential to control the root knot nematodes and other pests of plants. In this study different concentrations of the solution extracted from neem cake was tested against the second stage juveniles of the plant parasitic nematode Meloidogyne spp. and four phytopathogenic fungi: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytopthora capsici. Toxicity of neem cake extractions is represented by the EC50 value for the second-stage juvenile (J2) of Meloidogyne spp. and the four phytopathogenic fungi via Probit analysis. A 5% dilution of the solvent extracting from neem cake already caused 100% larval mortality after 24 hours exposure. Undiluted neem cake extraction effectively inhibited the growth of the four phytopathogenic fungi. The EC50 value of neem cake on J2-larvae of Meloidogyne nematode and on the fungi Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici was 0.51, 0.74, 0.30, 0.51 and 4.33%, respectively
Bánh dầu neem là sản phẩm của quá trình ép nhân hạt neem để lấy dầu. Các hoạt chất sinh học từ dịch chiết bánh dầu neem đã được đánh giá có tiềm năng lớn trong phòng trừ tuyến trùng nốt sưng và các loài dịch hại khác của nhiều loại cây trồng. Trong nghiên cứu này các nồng độ dịch chiết khác nhau của bánh dầu neem đã được thử nghiệm khả năng diệt tuyến trùng (ấu trùng tuổi 2 thuộc giống Meloidogyne spp.) và ức chế 4 loài nấm bệnh như: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. và Phytopthora capsici. Độc tính của dịch chiết bánh dầu neem được biểu diễn bởi giá trị EC50 đối với ấu trùng tuổi 2 của tuyến trùng Meloidogyne spp. và các loài nấm bệnh thông qua phân tích Probit. Dịch chiết bánh dầu neem ở nồng độ 5% đã làm chết 100% cá thể IJ2 của Meloidogyne spp sau 24 giờ phơi nhiễm. Dịch nguyên chất bánh dầu neem ức chế cả 4 loài nấm bệnh. Giá trị EC50 của bánh dầu neem lên ấu trùng tuổi 2 của Meloidogyne spp và các loài nấm bệnh Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici tương ứng là 0.51, 0.74, 0.30, 0.51 và 4.33%

Neem cake is a product of the cold pressing from the neem kernels to obtain neem oil. Bio-active substances from neem cake extracted solutions were evaluated for their potential to control the root knot nematodes and other pests of plants. In this study different concentrations of the solution extracted from neem cake was tested against the second stage juveniles of the plant parasitic nematode Meloidogyne spp. and four phytopathogenic fungi: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytopthora capsici. Toxicity of neem cake extractions is represented by the EC50 value for the second-stage juvenile (J2) of Meloidogyne spp. and the four phytopathogenic fungi via Probit analysis. A 5% dilution of the solvent extracting from neem cake already caused 100% larval mortality after 24 hours exposure. Undiluted neem cake extraction effectively inhibited the growth of the four phytopathogenic fungi. The EC50 value of neem cake on J2-larvae of Meloidogyne nematode and on the fungi Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici was 0.51, 0.74, 0.30, 0.51 and 4.33%, respectively.
Bánh dầu neem là sản phẩm của quá trình ép nhân hạt neem để lấy dầu. Các hoạt chất sinh học từ dịch chiết bánh dầu neem đã được đánh giá có tiềm năng lớn trong phòng trừ tuyến trùng nốt sưng và các loài dịch hại khác của nhiều loại cây trồng. Trong nghiên cứu này các nồng độ dịch chiết khác nhau của bánh dầu neem đã được thử nghiệm khả năng diệt tuyến trùng (ấu trùng tuổi 2 thuộc giống Meloidogyne spp.) và ức chế 4 loài nấm bệnh như: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. và Phytopthora capsici. Độc tính của dịch chiết bánh dầu neem được biểu diễn bởi giá trị EC50 đối với ấu trùng tuổi 2 của tuyến trùng Meloidogyne spp. và các loài nấm bệnh thông qua phân tích Probit. Dịch chiết bánh dầu neem ở nồng độ 5% đã làm chết 100% cá thể IJ2 của Meloidogyne spp sau 24 giờ phơi nhiễm. Dịch nguyên chất bánh dầu neem ức chế cả 4 loài nấm bệnh. Giá trị EC50 của bánh dầu neem lên ấu trùng tuổi 2 của Meloidogyne spp và các loài nấm bệnh Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici tương ứng là 0.51, 0.74, 0.30, 0.51 và 4.33%.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico
O cajueiro tem uma grande importÃncia sÃcio-econÃmica e cultural para a regiÃo Nordeste, por gerar emprego para os agricultores, sendo por muitas vezes a sua Ãnica fonte de renda. Assim, as pesquisas com esta fruteira tÃm aumentado bastante nos Ãltimos anos, visando primariamente, reduzir os danos fitossanitÃrios e elevar a produtividade. As doenÃas, como nas demais espÃcies cultivadas, causam significativos danos. Dentre as doenÃas do cajueiro, a resinose e a podridÃo preta das hastes, ambas causadas pelo fungo Lasiodiplodia theobromae, se destacando entre as principais doenÃas em algumas regiÃes do semi-Ãrido nordestino. PorÃm, ainda hà poucos estudos sobre a biologia, morfologia, caracterizaÃÃo genÃtica desse patÃgeno, fato que dificulta o seu manejo. Este estudo objetivou caracterizar a diversidade genÃtica e patogÃnica de uma populaÃÃo com 22 isolados de L. theobromae associada ao cajueiro, utilizando marcadores microssatÃlites e inoculaÃÃes em plantas de cajueiro. Para a caracterizaÃÃo molecular, extraiu-se o DNA de cada isolado a partir do micÃlio do fungo crescido em meio lÃquido. Cada amostra foi submetida à reaÃÃo de polimerase em cadeia (PCR) com 15 pares de iniciadores desenvolvidos para flanquear regiÃes especificas contendo sequÃncias simples repetidas (microssatÃlites) dessa espÃcie. Os produtos amplificados foram observados em gel de eletroforese corados com brometo de etÃdio, e os dados tabulados em planilha binÃria e analisados pelo mÃtodo de agrupamento nÃo balanceado baseado na mÃdia aritmÃtica (UPGMA), utilizando o programa MVSP. As similaridades genÃticas foram estimadas pelo coeficiente de Nei e Li. Os resultados indicaram uma grande variabilidade genÃtica da populaÃÃo avaliada, representada por diversos grupos de similaridade variando de 22,2% a 100%. TrÃs destes isolados apresentaram entre si, 100% de similaridade genÃtica com os marcadores testados. No entanto, eram morfologicamente diferentes e um deles era procedente de localidade diferente que os demais. Os dados mostraram que a similaridade genÃtica por si sà nÃo deve ser aplicado como Ãnico fator de diversidade entre isolados. Para se testar a patogenicidade, os isolados foram inoculados em mudas de cajueiro, analisando-se o tamanho da lesÃo e a taxa de crescimento do fungo. Os resultados mostraram nÃo haver diferenÃas estatÃsticas entre os isolados, fato que demonstra que, independente do local de origem, todos os isolados mostraram-se patogÃnicos, e a sua taxa de crescimento decresce, isto Ã, a planta deve produzir algum tipo de defesa contra o patÃgeno, o que dificulta a sua infecÃÃo.
The cashew has a major socio-economic and cultural development for the Northeast region, by generating employment for farmers and is frequently their only source of income. Thus, research on this fruit has increased greatly in recent years, aimed primarily to reduce damage and increase plant productivity. Diseases, as in other crops, cause significant damage. Among cashew diseases, gummosis and black stem rot, both caused by the fungus Lasiodiplodia theobromae, stand out among the major diseases in some regions of the semi-arid. However, there are few studies on the biology, morphology, genetic characterization of this pathogen, a fact that burden their management. This study aimed to characterize the genetic and pathogenic diversity of a population of 22 isolates of L. theobromae associated with cashew, using microsatellite markers and inoculations in cashew plants. For molecular characterization, DNA was extracted from mycelium of each isolated of the fungus grown in liquid medium. Each DNA sample was subjected to a polymerase chain reaction (PCR) with 15 pairs of primers designed to flank specific regions containing simple repeated sequences (microsatellites) in this species. The amplified products were observed in gel electrophoresis stained with ethidium bromide, and the data recorded on a binary and analyzed by grouping unbalanced based on arithmetic mean (UPGMA) using the program MVSP. Genetic similarities were estimated by the coefficient of Nei and Li The results indicated a large genetic variability of the population, represented by different groups of similarity ranging from 22.2% to 100%. Three of these isolates showed 100% genetic similarity with the markers tested. However, they were morphologically differences and one of them was originated from different location than the others. The data showed that the genetic similarity alone should not be applied as the only factor of diversity among isolates. To test the pathogenicity, each isolate was inoculated on cashew seedlings. Lesion development size and recovery of the fungus were recorded. The results showed no statistical differences between the isolates, which demonstrates that, regardless of place of origin, all isolates were pathogenic, and its growth rate decreases, i. e, the plant must produce some form of defense against pathogen, which hinders their infection.

碩士
國立中興大學
植物病理學系所
101
Papaya (Carica papaya) is an economically important crop in Taiwan. In recent years, a bacterial disease, black rot of papaya, seriously occurred in central and southern Taiwan. Based on physiological, biochemical and pathogenicity tests, the causal agent was identified as Erwinia sp. The 16S rDNA sequences of these Erwinia sp. (EP) strains were 97.9% similarity to Erwinia papayae. In order to clarify the taxonomic status of these EP isolates from Taiwan, multilocus sequence analysis (MLSA) and biochemical tests were used to identify the EP strains from black rot of papaya in this study. In the results of biochemical tests, the EP strains could utilize D-mannitol and L-arabinose, but did not produce blue pigment on KB or YDC medium. However, Erwinia papayae produces blue pigment on KB madium and utilizes L-arabinose but not D-mannitol. E. mallotivora does not produce blue pigment and it utilizes D-mannitol but not L-arabinose. The results of physiological and biochemical tests can not effectively identify the taxonomic status of EP strains. Phylogenetic trees were constructed based on the sequences of the four housekeeping genes, gyrB, rpoD, atpD and infB. The four gene sequence of EP strains alignment with E. papayae and E. mallotivora up to 99% identity. Neighbour-joining (NJ), maximum-parsimony (MP) and maximum-likelihood (ML) phylogenies of gyrB, rpoD, atpD and infB genes were compared. The EP strains formed a separate cluster from the E. papayae and E. mallotivora with bootstrap value of 100%. While the EP strains groups on the border of E. papayae with bootstrap value of 78-93 %. The results of this study revealed that the caused agent of black rot of papaya in Taiwan were E. papayae. The MLSA data suggested that EP strains could be a novel species, phylogenetically related to E. papayae.

Dupla diplomação com a Université Libre de Tunis
Chestnut fruits are popular fruits commercialized as fresh or processed ready to be used as products. Because of its high moisture and suitable nutrient content, the chestnut kernel has the potential to support the growth of a wide spectrum of spoilage fungi. The major postharvest problems associated with stored chestnut fruits are rots, which result in major losses in fruit quality. Although the empirical knowledge determines a high level of rot in Portuguese chestnuts, there are no scientific studies on the incidence and severity of the problem, and the causal agents have never been identified. This knowledge is of the utmost importance and urgency for industry and retailers to correctly address the issue, in an effort to reduce yield loss due to rot. The aims of this work were: i) to determine the incidence, abundance and diversity of rots in three chestnut varieties of Trás-os-Montes – Judia, Longal and Martaínha – at different postharvest stages of storage and processing, and ii) to identify the main potential agents of rots. For this purpose chestnuts were internally and externally inspected for presence of damages, infestation and infection. Samples from variety Martaínha were identified as the most resistant to fungal growth, while samples from variety Longal were less resistant to fungal growth and infestation. A high diversity of species has been molecularly identified by sequencing the ITS region: 37 different species belonging to 16 genera. The dominant fungal species found with high frequency were Mucor racemosus f. sphaerosporus (24.2% of frequency), Penicillium brevicompactum (16.7%) and Penicillium thomii (causal agents of green rot; 13.9%), Ciboria batschiana (the causal agent of black rot; 10.6%) and Botrytis cinerea (the causal agent of grey rot10.6%). Gnomoniopsis smithogilvyi, the causal agent of brown rot, was also identified with a frequency of 6.4%. The results show that the causal agents of various chestnut rots already identified in other countries are also present in Portuguese nuts. Studies must follow with the aim of developing control measures against the identified rot-causing fungi.
A castanha é um fruto popular comercializado e consumido tanto em fresco como processado. Devido à sua elevada humidade e teor de nutrientes, a castanha tem o potencial de permitir o crescimento de um amplo espectro de fungos de deterioração. Os principais problemas pós-colheita associados à castanha armazenada são as podridões, que resultam em grandes perdas na qualidade dos frutos. Embora o conhecimento empírico evidencie um alto nível de podridão nas castanhas portuguesas, não existem estudos científicos sobre a incidência e gravidade do problema, e os agentes causais nunca foram identificados. Esse conhecimento é da maior importância e urgência para a indústria e os comerciantes resolverem o problema adequadamente, no sentido de reduzirem as perdas de rendimento devido à podridão. Os objetivos deste trabalho foram: i) determinar a incidência, abundância e diversidade de podridões em três variedades de castanha de Trás-os-Montes - Judia, Longal e Martaínha - em diferentes estágios pós-colheita de armazenamento e processamento; e ii) identificar os principais agentes causais das podridões. Para esse fim, as castanhas foram inspecionadas interna e externamente quanto à presença de danos, infestações e infeções. Amostras da variedade Martaínha foram identificadas como as mais resistentes ao crescimento de fungos, enquanto amostras da variedade Longal foram menos resistentes ao crescimento e infestação de fungos. Uma alta diversidade de espécies de fungos foi identificada molecularmente através da região ITS: 37 espécies diferentes pertencentes a 16 géneros. As espécies dominantes encontradas com alta frequência foram Mucor racemosus f. sphaerosporus (24.2% de frequência), Penicillium brevicompactum (16.7%) e Penicillium thomii (13.9%) (agentes causais de podridão verde), Ciboria batschiana (agente causal da podridão negra; 10.6%) e Botrytis cinerea (agente causal da podridão cinzenta; 10.6%). Gnomoniopsis smithogilvyi, agente causal da podridão castanha, também foi identificado, com frequência de 6.4%. Estes resultados mostram que os principais agentes causais das podridões da castanha se encontram também nas castanhas portuguesas. Estão em curso estudos para desenvolvimento de métodos de controlo des fungos nas castanhas.
This work was developed under the scope of the project “ValorCast - Valorização da castanha e otimização da sua comercialização”, ref. PDR2020-101-032030. Funded by Fundo Europeu Agrícola de Desenvolvimento Rural (FEADER) and Portuguese Government, under the scope of Ação 1.1 «Grupos Operacionais», Medida 1. «Inovação», PDR 2020 – Programa de Desenvolvimento Rural do Continente.

碩士
國立屏東科技大學
生物科技系所
105
Black rot disease (BRD) caused by Xanthomonas campestris pv. campestris (Xcc) is considered the most important disease of crucifers worldwide. This destructive disease causes considerable yield losses up to 50% by premature defoliation. Various strategies have been developed to control BRD on cabbage, including the use of antagonistic microorganisms. In this study, we attend to assay the possibility of applying Bacillus amyloliquefaciens strains, PMB04 and PMB05, to control BRD on cabbage. Firstly, we assay the inhibitory effect of B. amyloliquefaciens strains againsts Xcc strains on nutrient agar plate. The inhibitory assay revealed that both antagonistic strains were able to inhibit the growth of Xcc strains. Moreover, PMB04 exhibited strongerinhibitory effect against two Xcc strains. These two antagonistic strains were further used to evaluate their biocontrol efficacy of BRD by two seed coating strategies, including preventing and scavenging treatment. As a result, both two treatments exhibited that BRD on cabbage was significantly suppressed by two B. amyloliquefaciens strains. Above all, PMB05 showed stronger suppression than PMB04 in both two seed coating strategies. On the contrary, the population of Xcc in plants was strongly reduced by PMB04 rather than PMB05. Our previous studies showed that B. amyloliquefaciens PMB05 was able to intensify plant immune system for further enhancing disease resistance of crops. To gain more insight on this response, we observed the deposition of callose to evaluate the activation of PAMP-triggered immunity. Correspondingly, callose deposition on the leaf of cabbage was strongly intensified by the cells of PMB05 upon the inoculation of Xcc62 or Xcc64. In addition, the growth of cabbage was not improved by the application of B. amyloliquefaciens. To optimize the biocontrol efficacy of BRD by PMB05, the fermentation liquid of PMB05 was further applied. Results exhibited that disease incidence and disease severity were completely suppressed. In conclusion, BRD could be controlled by B. amyloliquefaciens strain PMB04 due to the antagonistic effect. Most importantly, plant immune response intensified by B. amyloliquefaciens strain PMB05 and its fermentation liquid provides strong efficacy against BRD. The strong biocontrol efficacy might be due to the intensification of plant immunity by PMB05. We concluded that the application of these two B. amyliliquefaciens strains to control BRD on cabbage would be promising in the future.