Universiẗat, Diss., 2004--Bonn.

Τα υγρά παραπροϊόντα των ελαιοτριβείων (Olive Mill Wastewater, OMW) ή αλλιώς ο κατσίγαρος αποτελούν το κύριο παραπροϊόν της παραγωγικής διαδικασίας του ελαιόλαδου. Αποτελούν έναν από τους σημαντικότερους παράγοντες περιβαλλοντικής ρύπανσης, όπου οι τεράστιες επιπτώσεις τους οφείλονται κυρίως στα φυσικοχημικά τους χαρακτηριστικά. Αν και η απόρριψη ή η ενσωμάτωση των υγρών παραπροϊόντων στο έδαφος αποτελεί μια εναλλακτική μέθοδο επεξεργασίας και αξιοποίησης τους, εντούτοις οι αντιμικροβιακές και φυτοτοξικές ιδιότητες τους δυσχεραίνουν την υλοποίηση της. Παράλληλα όμως η εκμετάλλευση ορισμένων ιδιοτήτων τους συντελεί στη διερεύνηση πρωτότυπων, εναλλακτικών και οικολογικών τρόπων καταπολέμησης διαφόρων φυτοπαθογόνων μικροοργανισμών. Σύμφωνα με την πλειονότητα των βιβλιογραφικών αναφορών τα παραπροϊόντα αυτά είναι πλούσια σε οργανικά και ανόργανα συστατικά ενώ η βασική αιτία της τοξικής τους δράσης είναι η μεγάλη περιεκτικότητα τους σε φαινολικές ενώσεις. Στην παρούσα μεταπτυχιακή διατριβή έγινε προσπάθεια εκτίμησης της δράσης τους ενάντια στο φυτοπαθογόνο μύκητα Sphaerotheca fusca με απώτερο σκοπό να διερευνηθεί η όποια φυτοπροστατευτική τους δράση σε φυτά αγγουριού και η συγκέντρωση η οποία θα παρουσίαζε τη μεγαλύτερη αποτελεσματικότητα χωρίς ταυτόχρονα να προκαλεί φυτοτοξικές αντιδράσεις. Συγκεκριμένα πραγματοποιήθηκε μια σειρά in vitro βιοδοκιμών για τον έλεγχο της φυτοτοξικότητας των υγρών παραπροϊόντων στη βλάστηση σπερμάτων αγγουριού και της εξέλιξης της βλάστησης των κονιδίων του μύκητα, ενώ παράλληλα διεξήχθησαν in vivo και in planta πειράματα για τη μελέτη της εκλεκτικότητας των διαφόρων συγκεντρώσεων των παραπροϊόντων και της ελάχιστης αποτελεσματικής δόσης τους υπό συνθήκες αγρού. Σύμφωνα με τα αποτελέσματα του ελέγχου φυτοτοξικότητας, τα υγρά παραπροϊόντα των ελαιοτριβείων παρουσίασαν έντονη φυτοτοξικότητα ως πυκνό διάλυμα, ενώ η δράση τους περιορίστηκε κυρίως στην επιμήκυνση και τη ζωτικότητα του ριζιδίου. Συγχρόνως προκάλεσαν μια καθυστέρηση τεσσάρων περίπου ημερών στη βλάστηση των σπερμάτων ενώ όσον αφορά τις διάφορες αραιώσεις που δοκιμάστηκαν, διαπιστώθηκε ότι δεν είχαν καμία φυτοτοξική επίδραση ή αντιθέτως η επίδραση τους υπήρξε θετική, προάγοντας την επιμήκυνση του ριζιδίου. Μικροσκοπικές παρατηρήσεις έδειξαν ότι το πυκνό διάλυμα των υγρών παραπροϊόντων παρεμπόδισε την ανάπτυξη του μύκητα για μια περίοδο περίπου μιας εβδομάδας, όταν εφαρμόστηκε μια μέρα πριν την τεχνητή μόλυνση και κάτω από συνθήκες υψηλής πίεσης μολύσματος. Η μείωση του ποσοστού των βλαστημένων κονιδίων και της ανάπτυξης των πρωτογενών και δευτερογενών υφών των κονιδίων και του μυκηλίου αποτελούν προκαταρτικές αποδείξεις της μυκητοστατικής δράσης των υγρών παραπροϊόντων. Τα αποτελέσματα των in vivo και in planta πειραμάτων, τα οποία συνάδουν με αυτά των in vitro βιοδοκιμών που αφορούν την εξέλιξη της βλάστησης των κονιδίων του μύκητα, έδειξαν ότι η προληπτική εφαρμογή των υγρών παραπροϊόντων προκάλεσε την μείωση στην ένταση της ασθένειας όπως επίσης την καθυστέρηση στην έναρξη της, εμφανίζοντας είτε άμεση δράση στα κονίδια του μύκητα είτε έμμεση μέσω του ξενιστή ενεργοποιώντας μηχανισμούς ανοχής των φυτών σε παθογόνα. Συγκρίνοντας τη δράση τους με αυτή του μάρτυρα, αποδείχθηκε ότι η εφαρμογή τους δεν είχε καμία επίδραση στο ρυθμό ανάπτυξης της ασθένειας. Μεγαλύτερη αποτελεσματικότητα εμφάνισε το πυκνό διάλυμα των υγρών παραπροϊόντων και ειδικότερα μετά από την εφαρμογή εβδομαδιαίων επεμβάσεων. Αξίζει να σημειωθεί ότι η δράση του, κάτω από συνθήκες μειωμένης έντασης μολύσματος, ήταν παρόμοια με αυτήν του θειούχου μυκητοκτόνου Thiovit®. Στα πλαίσια λοιπόν της συγκεκριμένης μεταπτυχιακής διατριβής αποδεικνύεται η προστατευτική δράση των υγρών παραπροϊόντων σε φυτά αγγουριού έναντι του μύκητα Sphaerotheca fusca, η οποία μάλιστα συνοδεύεται από μη φυτοτοξικές επιδράσεις στα υπέργεια όργανα των φυτών. Παρ’όλα αυτά για τη διερεύνηση του ακριβή μηχανισμού ή μηχανισμών δράσης των υγρών παραπροϊόντων, όπως επίσης την εξακρίβωση της εμπλοκής και του ρόλου των φαινολικών ουσιών στη δράση τους, κρίνεται αναγκαία περαιτέρω έρευνα / Olive mill wastewaters (OMW) are the main by-products of olive oil production. Due to their physicochemical characteristics, these wastes constitute one of the major environmental problems. The disposal and the application of OMW to the soil is an alternative method of waste process and utilization. The phytotoxic and antimicrobial properties of OMW, however, prevent the implementation of this method. At the same time, some of the OMW properties contribute to the development of novel and alternative disease-control strategies, with an ecological basis. According to the scientific literature, these wastes contain organic, inorganic and toxic compounds, such as phenolics. The current study aims at the evaluation of olive OMW antifungal activity against Sphaerotheca fusca. The overarching purpose is to investigate the efficacy of OMW in protecting cucumber against powdery mildew, in relation to their concentration and phytotoxicity. Particularly, in vitro bioassays were carried out in order to examine OMW phytotoxicity on cucumber seed germination and their effect on germination progress of S. fusca conidia. In vivo and in planta testings were carried out both to investigate the selectivity of OMW concentrations, and the minimal effective dose in field conditions. OMW showed strong phytotoxicity as undiluted solution. Their activity was restricted to the elongation and growth of the root. OMW caused delay in seed germination by four days, whereas no phytotoxicity reaction occurred at any of the dilutions. In contrast, their activity in a certain dilution was positive, inducing root elongation. Microscopic observations showed that pretreatment of cucumber plants with undiluted OMW, followed by artificial inoculation and under low levels of inoculums, inhibited fungal growth for a week. Both the reduction in conidial germinated rate and the reduction in primary, secondary hyphae and mycelium growth, constitute preliminary evidence of OMW fungistatic activity. The results of in vivo and in planta treatments, which are in accordance with these microscopic observations, indicate that a prophylactic application of OMW reduced the disease severity and caused a delay in disease onset. OMW exerted either a direct effect on the fungal conidia, or acted indirectly, by activating mechanisms of plant tolerance. Despite the delay in disease onset, the rate of disease development was similar compared to the control. OMW were more effective when applied without any prior dilution and especially after a 7-day interval between applications. It is noteworthy that OMW were nearly as effective as sulphur (Thiovit®), under low levels of inoculums. The current study has demonstrated the protective action of OMW against cucumber powdery mildew (Sphaerotheca fusca), followed by the lack of phytotoxic effects on upper plant organs. Further research would be useful to identify the exact mechanism(s) of OMW activity, as well as the role of phenolic compounds in the process.

The purpose of this study was to evaluate the ultrastructural aspects of the host-pathogen relationship between the fungus Sphaerotheca pannosa (Wallr.) Lev. and a susceptible horticultural variety of floribunda rose "Eutin". Attention was given to the penetration of hyphal tubes, and their subsequent development into: haustorial elements in epidermal cells of rose. In addition, changes in the morphology of infected epidermal cells were described. Thin sections of rose leaves infected with S. pannosa were examined in a Hitachi electron microscope, model HS-7, after glutaraldehyde-acrolein fixation, OsO4-uranyl acetate staining, and embedding in Epon plastic. The components of S. pannosa haustoria consisted of a haustorial body with several lobes. The body, and the lobes, were irregular in shape, and were characteristically surrounded by thin haustorial walls. Small amounts of sheath enclosed the haustorial walls, or was found lacking around some haustorial lobes. A slender neck attached the haustorium to the surface hyphal cell, and was considered an extension of that cell, and not the haustorium. At the base of the neck, a septum which contained a central pore, separated the haustorium from the surface hyphal cell, making the haustorium an independent cell. The haustorial protoplasts were similar to hyphal cells on the surface. They contained mitochondria, ribosomes, endoplasmic reticulum, lipid bodies, and vacuoles. Muclei were not detected. Penetration of the epidermal wall of rose by hyphal tubes was accompanied by cellular proliferations on the inner surfaces of the host wall. These proliferations showed layering, and some formed collars around the penetrating tubes. Although the patterns of the proliferations differed from the normal wall layers of the host, one of the layers of the collar exhibited strands of microfibrils. Some haustoria were completely surrounded by cellular proliferations which were similar in electron density to collar material. It was thought that these proliferations were extensions of the collar which formed in response to breakdown of parasitic control. It appeared that the plasma membrane of the host was the sheath membrane, and that it also lined the cavity of the collar.

MSCAGR / Department of Plant Production

The use of biological control agents (BCAs) in the past has shown limited success as its application has often been done incorrectly, and in addition, management practices are rarely altered to incorporate BCAs. Criteria for the correct application of BCAs have been devised as part of the research, and companies selling these products may use the said criteria. Such application will ensure the correct BCAs are used and, more specifically, used under the correct conditions. The powdery mildew (PM) fungus is often seen to develop resistance to synthetic fungicides and, therefore, alternative control measures are required. BCAs as an alternative pose less risk to the environment, workers and the consumer. A pre-trial has been conducted with a range of BCAs to see if they can control powdery mildew (PM) in a greenhouse environment on hydroponically grown cucumber (Cucumis sativus L.) plants using the variety Baccara that has only a moderate tolerance to PM. The BCAs have been compared to the control (synthetic fungicide: Bravo). Comparative work includes Coyier's model, which has been modified and adapted for these trials to determine the percentage of leaf area covered by the PM infection. Furthermore, the number of fruit harvested per treatment, kilogram yield, total mass of yield and average fruit mass is also used to determine the efficacy of the BCAs as these factors have economic significance to commercial growers. The pre-trial showed promise until the fertigation computer failed, resulting in a nutrient shortage and imbalance, confirming that BCAs alone cannot control PM. Synthetic fungicides were applied until control of PM and plant nutrition was regained. BCAs were re-introduced and used until the end of crop production. The confirmation from the pre-trial has led to the inclusion of silicon in conjunction with the BCAs in the two subsequent trials (Trials 1 & 2). Silicon was applied with the BCAs as a foliar spray on a weekly basis. In trials 1 and 2, the cucumber variety, Palladium, with a high genetic tolerance to PM is used, as this variety is suited to form part of the holistic approach used for trials 1 and 2. Trial 1 showed that treatment A, containing Streptomyces griseovirdis and Streptomyces aureofaciens, had the highest yield. Both of these are bacterial BCAs and demonstrated their adaptability to varied climatic conditions, notably when low humidity was experienced. In treatment B, Trichoderma harzianum strains, Rifai and Uppington, show the slowest rate of PM development. In trials 1 and 2, the best actual PM control was obtained by two fungal based BCAs (Trial 1, treatment C was Ampelomyces quisqualis) and (Trial 2, treatment B was Trichoderma harzianum strains, Rifai and Uppington), showing that fungal BCAs have a place for this application, but the growth-enhancing properties of bacterial based BCAs make economic sense and would make them attractive to growers. Treatment A (Streptomyces spp.) had the most number of fruit for the entire growing period and the best overall yield (kg yield) again. Two of the BCA / silicon treatments have marginally better PM control compared to that of the control (E) treatment, although not statistically significant. Treatment E (control) has the highest average fruit mass in this instance but does not have the highest yield (kg yield) when compared to treatments A and B, possibly due to the growth-enhancing properties of most of these BCAs. Therefore, most of these BCA treatments give fairly inconsistent results that vary possibly according to season, humidity and temperature, making it difficult to predict their efficacy. Using combinations or weekly alternations of these BCAs with extremes of climatic adaptation will probably be the most reliable method of obtaining consistent results. Bacterial BCAs are shown to have lower humidity requirements and produce the most consistent results in terms of fruit number, yield and fruit mass and a combination of bacterial and fungal based BCAs would possibly be the best as this would control PM and yet still have the growth enhancing properties from the bacterial based BCAs. From the research, it can be said that some BCAs in trials 1 and 2 produce results similar to that of the control in terms of percentage leaf area covered by PM and some are shown to have improved yields. Results produced from certain BCA treatments are thus equal to the control; yet provide an environmentally friendly alternative to synthetic fungicides. Silicon is listed as a beneficial element rather than an essential element; however, literature claims it to be highly effective in treating PM in cucurbits. Results from trials 1 and 2 show that control of PM is possible in most cases, when a holistic approach is used. This approach includes a cucumber variety with a high PM tolerance, optimum nutrition, cultural practices and silicon in combination with the BCAs. A complete change of management practices is necessary to implement such a BCA program. / Agriculture, Animal Health & Human Ecology / M. Tech. (Nature Conservation)

The use of biological control agents (BCAs) in the past has shown limited success as its application has often been done incorrectly, and in addition, management practices are rarely altered to incorporate BCAs. Criteria for the correct application of BCAs have been devised as part of the research, and companies selling these products may use the said criteria. Such application will ensure the correct BCAs are used and, more specifically, used under the correct conditions. The powdery mildew (PM) fungus is often seen to develop resistance to synthetic fungicides and, therefore, alternative control measures are required. BCAs as an alternative pose less risk to the environment, workers and the consumer. A pre-trial has been conducted with a range of BCAs to see if they can control powdery mildew (PM) in a greenhouse environment on hydroponically grown cucumber (Cucumis sativus L.) plants using the variety Baccara that has only a moderate tolerance to PM. The BCAs have been compared to the control (synthetic fungicide: Bravo). Comparative work includes Coyier's model, which has been modified and adapted for these trials to determine the percentage of leaf area covered by the PM infection. Furthermore, the number of fruit harvested per treatment, kilogram yield, total mass of yield and average fruit mass is also used to determine the efficacy of the BCAs as these factors have economic significance to commercial growers. The pre-trial showed promise until the fertigation computer failed, resulting in a nutrient shortage and imbalance, confirming that BCAs alone cannot control PM. Synthetic fungicides were applied until control of PM and plant nutrition was regained. BCAs were re-introduced and used until the end of crop production. The confirmation from the pre-trial has led to the inclusion of silicon in conjunction with the BCAs in the two subsequent trials (Trials 1 & 2). Silicon was applied with the BCAs as a foliar spray on a weekly basis. In trials 1 and 2, the cucumber variety, Palladium, with a high genetic tolerance to PM is used, as this variety is suited to form part of the holistic approach used for trials 1 and 2. Trial 1 showed that treatment A, containing Streptomyces griseovirdis and Streptomyces aureofaciens, had the highest yield. Both of these are bacterial BCAs and demonstrated their adaptability to varied climatic conditions, notably when low humidity was experienced. In treatment B, Trichoderma harzianum strains, Rifai and Uppington, show the slowest rate of PM development. In trials 1 and 2, the best actual PM control was obtained by two fungal based BCAs (Trial 1, treatment C was Ampelomyces quisqualis) and (Trial 2, treatment B was Trichoderma harzianum strains, Rifai and Uppington), showing that fungal BCAs have a place for this application, but the growth-enhancing properties of bacterial based BCAs make economic sense and would make them attractive to growers. Treatment A (Streptomyces spp.) had the most number of fruit for the entire growing period and the best overall yield (kg yield) again. Two of the BCA / silicon treatments have marginally better PM control compared to that of the control (E) treatment, although not statistically significant. Treatment E (control) has the highest average fruit mass in this instance but does not have the highest yield (kg yield) when compared to treatments A and B, possibly due to the growth-enhancing properties of most of these BCAs. Therefore, most of these BCA treatments give fairly inconsistent results that vary possibly according to season, humidity and temperature, making it difficult to predict their efficacy. Using combinations or weekly alternations of these BCAs with extremes of climatic adaptation will probably be the most reliable method of obtaining consistent results. Bacterial BCAs are shown to have lower humidity requirements and produce the most consistent results in terms of fruit number, yield and fruit mass and a combination of bacterial and fungal based BCAs would possibly be the best as this would control PM and yet still have the growth enhancing properties from the bacterial based BCAs. From the research, it can be said that some BCAs in trials 1 and 2 produce results similar to that of the control in terms of percentage leaf area covered by PM and some are shown to have improved yields. Results produced from certain BCA treatments are thus equal to the control; yet provide an environmentally friendly alternative to synthetic fungicides. Silicon is listed as a beneficial element rather than an essential element; however, literature claims it to be highly effective in treating PM in cucurbits. Results from trials 1 and 2 show that control of PM is possible in most cases, when a holistic approach is used. This approach includes a cucumber variety with a high PM tolerance, optimum nutrition, cultural practices and silicon in combination with the BCAs. A complete change of management practices is necessary to implement such a BCA program. / Agriculture, Animal Health and Human Ecology / M. Tech. (Nature Conservation)