Aigner, Elizabeth Jo.
Thesis (M.S.)--Youngstown State University, 1996. / Includes bibliographical references (p. 86-90).
Determination of the effects of sunlight and UV irradiation on the structure, viability and reapplication frequency of the biopesticide cryptophlebia leucotreta granulovirus in the protection against false codling moth infestation of citrus cropsMwanza, Patrick January 2015 (has links)
Cryptophlebia leucotreta granulovirus (CrleGV-SA) is a baculovirus specifically pathogenic to the citrus pest false codling moth, Thaumatotibia leucotreta. CrleGV- SA is formulated as a commercial biopesticide, Cryptogran® (River Bioscience, South Africa). The virus has a stable, proteinaceous crystalline occlusion body (OB) that protects the nucleocapsid. The major limitation to the use of baculoviruses is their susceptibility to the ultraviolet (UV) component of sunlight, which rapidly and greatly reduces their efficacy as biopesticides. The UVA and UVB components are the most destructive to biological organisms. To date no publication has reported the effect of UV on the structure and virulence of CrleGV, or the effectiveness of the OB as a UV protectant. In this study the effect of UV irradiation on the structure and infectivity of pure CrleGV-SA and Cryptogran® was investigated using scanning electron microscopy (SEM), Raman spectroscopy, qPCR, and bioassays. The project included laboratory and field studies. In the laboratory, CrleGV-SA and Cryptogran® were exposed to either UVA or UVB for periods of 24 hours to 7 days before analysis. In the field, Cryptogran® was applied to trees in a citrus orchard with young fruit. The fruit were collected from 24 hours to 28 days after application and bioassays conducted to assess the effect of sunlight over time on virus structure and efficacy when applied to the northern or southern sides of the trees. No surface morphological changes to the virus were detected using SEM. However, small compositional changes were detected by Raman spectroscopy. qPCR and bioassays demonstrated that UV irradiation damaged the viral DNA, greatly reducing the infectivity of pure CrleGV-SA and Cryptogran®. Exposure to UVB reduced the virulence of the virus more than UVA. The field studies revealed that the activity of CrleGV-SA decreased more on the northern side of the trees than on the southern side.
Georgiou, Kathy Hadje
16 January 2012
MSc., Faculty of Science, University of the Witwatersrand, 2011 / The 2-isopropenyl-2,3-dihydrobenzofuran moiety is found in many naturally occurring compounds including rotenone, a complex pentacyclic molecule isolated from several leguminous plants of the Derris and Lonchocarpus species. Interest in rotenone stems from the fact that it possesses significant pesticidal and piscicidal properties which have been employed for centuries. Furthermore, as it has three stereogenic centres, rotenone poses an interesting and challenging synthetic target for organic chemists. Although various syntheses of this natural compound have been reported, none of these were stereoselective. The first stereoselective total synthesis of rotenone is described in this dissertation. Initially, a model study was conducted in which the simplest of the natural rotenoids, munduserone, was synthesised. The key step in this transformation involves the use of a platinum catalysed 6-endo-hydroarylation reaction of an alkynone intermediate, thus affording munduserone in 6 steps and an overall yield of 23%. We then attended to the synthesis of the more complex rotenoid, rotenone. Rotenone was synthesised by the initial assembly of a chiral (-)-(R)-2-isopropenyl-2,3-dihydrobenzofuran-4-ol moiety, asymmetrically accessible using a stereoselective Pd π-allyl mediated cyclisation of (E)-4-(2,6-dihydroxyphenyl)-2-methylbut-2-enyl methyl carbonate. Having constructed the dihydrobenzofuran in an enantiomeric excess of 94.8%, the chromene part of rotenone could then be synthesised. To this end, the LDA mediated coupling reaction of the formylated dihydrobenzofuran and 1,2-dimethoxy-4-(prop-2-ynyloxy)benzene, gave a secondary alcohol which was subsequently oxidised to the corresponding alkynone, (-)-(R)-(6,7-dimethoxy-2H-chromen-4-yl)(4-methoxy-2-isopropenyl-2,3-dihydrobenzofuran-5-yl)methanone. A 6-endo-hydroarylation reaction was employed as a mild strategy to construct the chromene moiety, (-)-(R)-(6,7-dimethoxy-2H-chromen-4-yl)(4-methoxy-2-isopropenyl-2,3-dihydrobenzofuran-5-yl)methanone. Finally, a deprotection and a base-catalysed intramolecular oxo-Michael addition concluded the first stereoselective synthesis of rotenone in 17 steps and an overall yield of 0.02%
Testing for microbiologically active compounds extracted from members of the family laminaceae and other indigenous plants.Gurlal, Prenitha. January 2005 (has links)
The Labiatae is a large family that occurs worldwide and have species that are adapted to almost all habitats and altitudes. Plectranthus is in this family. Plectranthus species are beautiful South African shrubs. The genus Plectranthus belongs to subfamily Nepetoideae of tribe Ocimeae. The test microorganisms were chosen carefully as each one belonged to a different taxonomic group of fungi and bacteria. Biologically active mono- and sesquiterpenoids are frequently found in many species of Plectranthus but there are little published data that directly link the presence of specific compounds in a species with the traditional uses of that species. Various Plectranthus spp. were collected and dried, followed by chemical extraction using various solvents. Dichloromethane extracts of P. fruticosus and P. ecklonii were screened for antibacterial and antifungal activities using the agar well and trench diffusion methods. It was found that both methods produced inconsistent results. The trench method required a bigger volume of plant extract to be filled into the well, hence, better biological activity was observed with that method. The well method required a smaller volume therefore poor activity was noted with this assay. The size of inhibition zones are dosage dependent. Overall, both plant extracts exhibited antibacterial but no antifungal properties. The pure compound (1), 11-Hydroxy-2-(4-hydroxybenzoyl)-5,7,9(11),13-abietatetraen-12-one, isolated from P. ecklonii was found to be the same as compound (10) which was isolated from P. lucidus. P. hadiensis was extracted using dichloromethane and hexane. The dichloromethane extract proved to contain much higher biological activity than the hexane extract. Three pure compounds, identified as diterpenes, were isolated from the crude dichloromethane extract of P. hadiensis. 6,7-Dihydroxyroyleanone-6,7,12-trihydroxy-8,12-abietadiene-ll,14-dione (2) and 7(alpha)-formoxy-6(beta)-hydroxyroyleanone (3) exhibited good antibacterial and antifungal activity but not against all the test organisms. The remaining pure compound, 7(alpha)-acetoxy-6(beta)hydroxyroyleanone (4), exerted good antifungal activity. This was measured by the inhibition zone which measured up to 14mm when compound 4 was tested against S. sclerotiorum. When testing the hexane extract against the Bacillus formulations, the pellets that were suspended once in Ringer's solution produced bigger inhibition zones than the pellets that were suspended twice. This could be due to bacterial cells washing out of the suspension. The dichloromethane extract of P. praetermissus proved to be very active against X campestris, producing an inhibition zone of 8 - 20mm. Two pure compounds were isolated from the crude extract and identified as diterpenes. Compound 5, 20(10--> 5)-abeo1( 10),6,8,11,13-abietapentaene-11,12,16-triol, and compound 6, 11,12,15-trihydroxy-20( 10-->5)-abeo-abieta-1-(10),6,8,11,13-pentaene are both known compounds which have previously been isolated from Salvia apiana. P. cilatus was extracted with chloroform and tested against various microorganisms for antifungal and antibacterial activities. It showed poor biological activity overall, except against S. sclerotiorum. The crude dichloromethane extract of P. zuluensis exhibited good antibacterial activity, which was limited to the Gram negative test organism. The extract produced an inhibition zone of 10-12mm when tested against X campestris. Pure compound 7, 2-hydroxy-4,6dimethoxyacetophenone, exerted excellent inhibition against B. subtilis and S. sclerotiorum. Neither compound 8, 1,2,4-trimethoxy-5-(2-propenyl)-benzene, nor compound 7, inhibited Candida spp., F. oxysporum and R. solani. Two diterpenes were isolated from the aerial plant parts of P. lucidus with dichloromethane and their structures elucidated by spectroscopic means. The pure compound 9, 11-hydroxy19-( 3-methyl-2-butenoyl)-5,7,9(11), 13-abietatetraen-12-one, showed moderate antifungal activity whereas compound 10, 11-hydroxy-2-(4-hroxybenzoyl)-5,7,9(11),13-abietatetraen12- one, showed high antifungal activity against R. solani, S. sclerotiorum and F. oxysporum. The crude and the pure compounds (formerly isolated from P. parviflorus) showed inhibition against X campestris. The dichloromethane extracts of P. purpuratus subsp. purpuratus and P. purpuratus subsp. tongaensis exhibit similar levels of biological activity when tested against the same test organisms. Poor antibacterial activity was noted with both extracts. However, excellent antifungal activity was depicted when both plant extracts were tested against F. oxysporum, R. solani and S. sclerotiorum. However, the highest biological activity was noted by R. solani which was totally inhibited by both dichloromethane extracts. The pure compound (11) isolated from P. purpuratus subsp. purpuratus was found to have the same chemical structure as compound (9) previously isolated from P. lucidus. The bioautography assay was used to detect and activity-guide the fractionation of antimicrobial compounds from all the Plectranthus spp. tested. The TLC fingerprint showed a zone of clearing around the lower bands of P. fruticosus and P. ecklonii when the plate was sprayed with a suspension of B. subtilis. This result is consistent with the agar well diffusion method. Clear zones were also noted on some bands of the extracts of P. zuluensis, P. ciliatus, P. hadiensis and P. praetermussis. Clear zones indicate inhibition of growth. Other plant extracts tested for biological activity were from the family Lamiaceae, however, not of the genus Plectranthus. Persicaria senegalensis, Pycnostachys reticulata and Ficus sur possessed moderate biological activity overall. It is interesting to note that P. senegalensis and F. sur exert high biological activity against Candida spp. This could be useful as herbal remedies for yeast infections. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.
The isolation, genetic characterisation and biological activity of a South African Phthorimaea operculella granulovirus (PhopGV-SA) for the control of the Potato Tuber Moth, Phthorimaea operculella (Zeller)Jukes, Michael David January 2015 (has links)
The potato tuber moth, Phthorimaea operculella (Zeller), is a major pest of potato crops worldwide causing significant damage to both field and stored tubers. The current control method in South Africa involves chemical insecticides, however, there is growing concern on the health and environmental risks of their use. The development of novel biopesticide based control methods may offer a potential solution for the future of insecticides. In this study a baculovirus was successfully isolated from a laboratory population of P. operculella. Transmission electron micrographs revealed granulovirus-like particles. DNA was extracted from recovered occlusion bodies and used for the PCR amplification of the lef-8, lef-9, granulin and egt genes. Sequence data was obtained and submitted to BLAST identifying the virus as a South African isolate of Phthorimaea operculella granulovirus (PhopGV-SA). Phylogenetic analysis of the lef-8, lef-9 and granulin amino acid sequences grouped the South African isolate with PhopGV-1346. Comparison of egt sequence data identified PhopGV-SA as a type II egt gene. A phylogenetic analysis of egt amino acid sequences grouped all type II genes, including PhopGV-SA, into a separate clade from types I, III, IV and V. These findings suggest that type II may represent the prototype structure for this gene with the evolution of types I, III and IV a result of large internal deletion events and subsequent divergence. PhopGV-SA was also shown to be genetically more similar to South American isolates (i.e. PhopGV-CHI or PhopGV-INDO) than it is to other African isolates, suggesting that the South African isolate originated from South America. Restriction endonuclease profiles of PhopGV-SA were similar to those of PhopGV-1346 and PhopGV-JLZ9f for the enzymes BamHI, HindIII, NruI and NdeI. A preliminary full genome sequence for PhopGV-SA was determined and compared to PhopGV-136 with some gene variation observed (i.e. odv-e66 and vp91/p95). The biological activity of PhopGV-SA against P. operculella neonate larvae was evaluated with an estimated LC₅₀ of 1.87×10⁸ OBs.ml⁻¹ being determined. This study therefore reports the characterisation of a novel South African PhopGV isolate which could potentially be developed into a biopesticide for the control of P. operculella.
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