京都大学 / 0048 / 新制・課程博士 / 博士(理学) / 甲第20752号 / 理博第4328号 / 新制||理||1622(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)教授 石田 厚, 教授 工藤 洋, 教授 田村 実 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DGAM

Fusarium oxysporum Schl. var. vanillae (Tucker) Gondon is known to cause root rot in Vanilla planifolia Andrews in most regions where it is grown, including the major plantations in Xishuangbanna, Yunnan Province of China. This is of serious economic concern to the Province since the vanilla flavouring extractable from the beans of the plant is a valuable food product and an important export commodity. There are no fungicides registered for the control of Fusarium root rot and the only available chemical control methods are ineffective and cause serious contamination of the soil. Breeding for resistance is difficult when no dominant gene is known or where little information is available on fungal pathogenicity. Biocontrol is the main alternative for disease control in this crop, an attractive approach because of increasing concerns for environmental protection. The investigation considers two biocontrol strategies: first the introduction of virulent, antagonistic, non-pathogenic strains, closely-related to the pathogen, to overcome pathogenic populations in infected soils; second the use of essential oils with antimicrobial properties when applied to infected soils. Pathogenicity tests have been done on 81 out of 87 F. oxysporum isolates collected in Yunnan Province. Among these, 32 isolates were non-pathogenic and 49 were pathogenic. The pathogenicity results showed the complexity of F. oxysporum in Yunnan. Seventeen isolates were recovered from the Daluo plantation, of which 14 were pathogenic isolates and 3 non-pathogenic isolates; 26 from the Menglun plantation, in which 12 were pathogenic and 14 were non-pathogenic; 18 isolates from the Manjingdai plantation, in which 12 isolates were pathogenic, whilst the other 6 were non-pathogenic and 20 were obtained from the plantation in Hekou i County, of which 11 were pathogenic isolates and 9 were non-pathogenic. Genetic diversity within this population of F. oxysporum has been investigated with respect to vegetative compatibility and to determine the relationship between VCGs and virulence. The VCG results showed that the 87 strains of Fusarium oxysporum f.sp vanillae isolated from Yunnan Province were complex. They could be distributed into 12 different VCGs and that a direct relationship between VCGs group and virulence could not be drawn. Two non-pathogenic strains, ML-5-2 and HK-5b-4-1, have been screened from 87 strains as candidate biocontrol agents by pathogenicity and VCG, which are self-incompatible and closely related to the pathogens. These two strains were effective in vanilla root rot control in controlled environments, but their effects in field experiments were less conclusive. Seven essential oils, which have long been regarded as having inhibitory effects on pathogens in nature, have also been investigated as biocontrol agents. Three oils, cinnamon oil, thyme oil and clove oil, were effective in inhibiting the growth of pathogen in vitro. These oils may develop into useful components of different management strategies with non-pathogenic strains. For the future, consideration will need to be given to the mechanism(s) of the interaction of the antagonistic components with the soil microbe population and host plant and also to appropriate formulation, to take account of soil type, crop status, cultural practices, environmental and economic factors. Biocontrol methods have considerable potential but must be acceptable to farmers as part of an overall crop management programme.

Dans la forêt de pin maritime (Pinus pinaster) des Landes de Gascogne (sud-ouest de France), la mortalité des pins causée par le champignon pourridié Armillaria ostoyae (Basidiomycète) a augmenté au cours des 30 dernières années. Les premiers cas de cette maladie ont été signalés quelques années après un changement majeur dans l'utilisation des terres, qui a eu lieu dans cette région suite au remplacement des landes et marais d'origine par une forêt plantée et gérée da façon intensive. Notre objectif était de comprendre les facteurs à l'origine de cette maladie émergente. Pour cela, nous avons étudié la distribution spatiale des dommages causés par le pathogène en relation avec des facteurs historiques, estimé la variabilité des traits fongiques liés au parasitisme et saprophytisme, et étudié l'histoire démographique d'A. ostoyae. La répartition actuelle de la mortalité induite par A. ostoyae est apparue dépendre de la présence des forêts préexistantes, ce qui suggère qu'A. ostoyae était fréquent dans ces zones forestières anciennes, qui ont agi comme un réservoir pour la colonisation des forêts plantées récentes. La production de rhizomorphes était significativement corrélée avec la virulence, suggérant que ce trait joue un rôle important dans le stade parasitaire d'A. ostoyae. Aucune relation significative entre le parasitisme et saprophytisme n'a été détectée, suggérant une absence de compromis évolutif entre ces traits. Enfin, le meilleur scénario démographique pour expliquer la structure de la population d'A. ostoyae dans la forêt des Landes est un scénario en deux étapes : il y aurait eu d'abord une diminution puis une expansion de la population fongique, qui semblait suivre la dynamique de la population d'hôtes. Le temps de génération d’A. ostoyae a été estimé entre 10 et 20 ans. / In the maritime pine (Pinus pinaster) forest of the Landes de Gascogne (south-western France), pine mortality due to the root rot fungus Armillaria ostoyae (Basidiomycete) has been increasing over the last 30 years. The first cases of this disease were reported a few years after a major change in land use which occurred in this region following the replacement of original moors by an intensively managed planted forest. Our aim was to understand the factors driving this disease emergence. For this, we investigated the spatial distribution of pathogen damage related to historical factors, estimated the variation in fungal traits related to parasitism and saprophytism and investigated the demographic history of A. ostoyae. The current distribution of A. ostoyae mortality appeared depending on the pre-existing forests, suggesting that A. ostoyae was commonly distributed in pre-existing forest areas which acted as a reservoir for the colonization of recent planted forests. The rhizomorphs production was significantly correlated with virulence, suggesting that this trait plays an important role in the parasitic stage of A. ostoyae, but no significant relationship between parasitism and saprophytism components was detected, which may suggest that there is no trade-off between these traits. Finally, the best demographic scenario to explain A. ostoyae population structure in the Landes forest is a two step scenario: there was first a decrease and then an expansion in the fungal population, which appeared to follow the dynamics of the host population. The generation time of A. ostoyae was estimated between 10 and 20 years.