Thesis (MSc)--University of Stellenbosch, 2011. / ENGLISH ABSTRACT: Plants are constantly exposed to biotic and abiotic stress inducing factors that threaten their existence. Biotic factors such as pathogens are the cause of huge yield losses to crop plants worldwide with fungal pathogens debatably constituting the worst damage. Fungal pathogens such as Botrytis cinerea, which has a wide host range, release cell wall degrading enzymes called endopolygalacturonases (ePGs) during plant infection. These ePGs break down the pectin component of the cell wall, thus providing an entry route, as well as nutrients for the fungus. Plants have evolved mechanisms to counteract and suppress the action of the ePGs. This is achieved through the action of cell wall associated proteins called polygalacturonaseinhibiting proteins, PGIPs. PGIPs directly inhibit ePGs and their inhibitory action also prolongs the existence of longer chain oligogalacturonide residues which are believed to elicit a cascade of defence responses. In grapevine, a PGIP encoding gene, VvPGIP1, was previously isolated and characterised. VvPGIP1, as well as nine non-vinifera grapevine PGIPs have been expressed in tobacco and shown to be potent antifungal proteins that caused the transgenic tobacco to have strong resistance phenotypes against Botrytis in whole plant infection assays. Following on the tobacco study, two of the non-vinifera PGIPs were expressed in cultivars of the susceptible Vitis vinifera. Characterisation of the putative transgenic population showed that transgene integration was successful, the transgenes were being expressed and there were at least 29 transgenic lines with independent integration events. The transgenic lines were confirmed to have active PGIPs (transgene-derived) in their leaves. Crude protein extracts from 22 lines exhibited 100% inhibition against crude B. cinerea PGs (BcPGs). The plant lines with positive transgene integration, expression, independent integration events and exhibiting 100% transgene-derived PGIP activity were further selected for whole plant and detached leaf antifungal assays where they were challenged with B. cinerea. The whole plant infection assay showed that expression of the non-vinifera PGIPs in V. vinifera promotes susceptibility to B. cinerea, not resistance. This surprising result could perhaps be explained by a quicker and stronger recognition between the pathogen and the host and the stronger activation of defence responses in the host. A more active hypersensitive response in the host would benefit Botrytis being a necrotroph. The type of lesions and the onset and speed of lesion development observed on the transgenics lines versus the wild type support this possibility. Knowledge gaps with regards to the efficiency of the ePG inhibition by the nonvinifera PGIPs during infection of grapevine tissue; the potential changes that might be caused by expressing PGIPs in a grapevine host with a native PGIP with high homology to the transgenes (including potential gene silencing) and the potential impact on defence signalling and defence responses all provides further avenues of study to elucidate this very interesting phenotype further. Overall, this study provides a comprehensively characterised population of transgenic plants that provides useful resources for in vivo analysis of PGIP function in defence, where the host plant harbours a native copy of the PGIP encoding gene. / AFRIKAANSE OPSOMMING: Plante word voortdurend blootgestel aan biotiese en abiotiese faktore, wat stres veroorsaak en hul bestaan bedreig. Biotiese faktore, soos patogene, veroorsaak groot verliese in wêreldwye gewasopbrengste, met swampatogene wat moontlik die grootste skade veroorsaak. Swampatogene, soos Botrytis cinerea, wat ‘n wye reeks gasheerplante kan infekteer, stel selwand-afbrekende ensieme tydens plantinfeksie vry, wat as endo-poligalakturonases (ePG’s). bekend staan. Hierdie ePG’s breek die pektienkomponent van die selwand af, wat gevolglik as ‘n ingangspunt dien,asook voedingstowwe vir die swam verskaf. Plante het meganismes ontwikkel om die aktiwiteit van hierdie ePG’s te bekamp en te onderdruk. Die aktiwiteit van die selwand-geassosieërde proteïene, genaamd poligalakturonase-inhiberende proteïene (PGIP’s), speel hier ‘n rol. PGIP’s inhibeer ePG’s direk en hul inhiberende aktiwiteit verleng ook die bestaan van langketting oligogalakturoniedresidu’s, wat blykbaar ‘n kaskade van weerstandsreaksies kan inisieer. ‘n PGIP-koderende geen, VvPGIP1, is voorheen uit wingerd geïsoleer en gekarakteriseer. VvPGIP1, asook nege nie-vinifera wingerd-PGIP’s is voorheen in tabak uitgedruk en bevestig as proteïene met sterk anti-swamaktiwiteit, soos bevestig deur die bevinding dat die transgeniese tabak ‘n weerstandsfenotipe teen Botrytis in heelplant-infeksietoetse het. Ná die tabakstudie is twee van die nie-vinifera PGIP’s uitgedruk in vatbare V. vinifera-kultivars. Karakterisering van die vermeende transgeniese bevolking het getoon dat die transgeen-integrasie suksesvol was, dat die transgeen uitgedruk word en dat daar ten minste 29 transgeniese lyne met onafhanklike integrasie gebeurtenisse geskep is. Daar is verder bevestig dat die transgeniese lyne aktiewe PGIP’s (transgeen-afkomstig) in hul blare het. Ongesuiwerde proteïenekstrakte van 22 lyne het 100% inhibisie teen ‘n mengsel van ongesuiwerde B. cinerea PGs (BcPGs) getoon. Die plantlyne met positiewe transgeenintegrasie en -uitdrukking, asook onafhanklike integrasiegebeure en wat 100% transgeen-afkomstige PGIP-aktiwiteit getoon het, is verder aan heel-plant en verwyderde blaarswaminfeksies met B cinerea onderwerp. Die heelplantinfeksietoetse het getoon dat uitdrukking van nie-vinifera PGIP’s in V. vinifera ‘n toename, in plaas van ‘n afname, in vatbaarheid teen B. cinerea veroorsaak. Hierdie verbasende resultaat kan moontlik toegeskryf word aan ‘n vinniger en sterker herkenningsreaksie tussen patogeen en gasheer en die moontlike sterker stimulering van weerstandsreaksies in die gasheer. ‘n Meer aktiewe hipersensitiewe reaksie in die gasheer sal tot die voordeel van Botrytis, wat ‘n nektrotroof is, wees. Die tipe letsel, asook die aanvang en spoed van letselontwikkeling wat waargeneem is in transgeniese lyne teenoor die wilde-tipe ondersteun hierdie moontlikheid. Gapings in kennis ten opsigte van die doeltreffendheid van die ePG-inhibisie deur die nievinifera PGIP’s tydens infeksie van wingerdweefsel, die moontlike veranderinge (insluitend ‘n moontlike geenuitdowingseffek) wat veroorsaak kan word deur die uitdrukking van PGIP-gene in ‘n kultivar met ‘n inheemse en baie homoloë PGIP-geen, kon ‘n invloed op weerstandseine en weerstandsreaksies gehad het. Hierdie aspekte lewer verdere studiemoontlikhede om hierdie interessante fenotipe verder te verklaar.Algeheel lewer hierdie studie ‘n breedvoeriggekarakteriseerde bevolking trangeniese plante, wat dien as nuttige hulpbronne vir in vivoanalise van PGIP se funksie in siekteweerstandbiedendheid, veral waar die gasheerplant ‘n inheemse kopie van die PGIP-koderende geen huisves.

Thesis (PhD)--University of Stellenbosch, 2003. / ENGLISH ABSTRACT: Grapevine leafroll-associated virus-3 is one of ten members of the C/osteroviridae that are known to infect grapevine. Nine of these viruses are associated with grapevine leafroll disease, of which GLRaV-1 and GLRaV-3 are the most important and widespread. Members of the C/osteroviridae are unique amongst the viruses, as it is the only known family whose members encode a heat shock protein 70 kOa homolog (Hsp70h). The Hsp70h is a movement protein (MP) that is required for the active translocation of the virion structure through the plasmodesmata into adjacent cells. Broad-spectrum resistance to unrelated viruses can be obtained by a pathogen-derived resistance (POR) strategy that is based on the expression of a dysfunctional MP in plants. The Hsp70h has two distinct domains. The N-terminal two thirds of the protein is an ATPase domain and shares high homology with the ATPase domains of all Hsp70h proteins from the C/osteroviridae and Hsp70 proteins from the prokaryote and eukaryote kingdoms. Conserved amino acids are found in the ATPase domain and are required for the positioning of the ATP at the catalytic site for ATP hydrolysis. The C-terminal domain is variable and the function of this domain in the Closteroviridae is not known. In prokaryote and eukaryote Hsp70 proteins, the C-terminal domain is required for protein-protein interactions. The American NY-1 isolate of GLRaV-3 has been sequenced and POR strategies have been attempted with the coat protein, divergent coat protein and replicase genes, but not with a dysfunctional form of the hsp70h gene. In this study, double-stranded RNA was isolated from a commercial vineyard with unknown virus status, but with distinct grapevine leafroll symptoms, and from two grapevine sources of known virus status, one with mild and one with severe symptoms. The GLRaV-3 hsp70h gene was amplified by RT-PCR from the dsRNA and the gene sequence was analysed. The hsp70h gene from the three virus sources contained more than 94% nucleotide sequence homology to the NY-1 isolate and the conserved amino acids required for ATPase activity were present. The hsp70h gene isolated from GLRaV-3 from a commercial Stellenbosch vineyard showing clear leafroll symptoms was selected for further work and was subjected to site-directed mutagenesis to engineer four point mutations in the gene. These four mutations resulted in the substitution of Asn for Asp", Gly for Thr1O, Lys for Glu 174 and Asn for Asp 197. The wild type (WT) and mutated (Mut) forms of the hsp 70h genes were cloned into a bacterial expression vector. Expression of both the WT- and Mut-Hsp proteins was achieved, and the protein was expressed in the insoluble inclusion bodies. All attempts to refold and isolate active proteins from the inclusion bodies were unsuccessful. Attempts to increase the concentration of soluble protein within the expressing bacteria were unsuccessful. Due to the lack of active protein, biochemical tests on the ATPase activity of the WT- and Mut-Hsp proteins could not be conducted. The wt- and mut-hsp genes were cloned into a plant expression vector for transformation into tobacco plants. These transformations were successful and gave rise to 22 Km' and 18 Km' plants from the WT- and Mut-Hsp constructs respectively. Two plant lines, M5 and M10, transformed with the mut-hsp transgene construct, appeared to have a high level of resistance to the challenging potato X potexvirus, whereas all the other tested plants were susceptible to the challenging virus. It was thus shown that a dysfunctional form of the GLRaV-3 Hsp70h could provide resistance to an unrelated virus in tobacco. / AFRIKAANSE OPSOMMING: Wingerdrolblaar-geassosieerde virus 3 (GLRaV-3) is een van 10 lede van die Closteroviridae wat wingerd kan infekteer. Nege van die virusse is met wingerdrolblaar geassosieer. Die GLRaV-1 en GLRaV-3 is die belangrikste en mees wyd verspreide lede van die rolblaar-geassosieerde Closteroviridae. Lede van die Closteroviridae is uniek in die opsig dat die virusse vir 'n 70 kDa-homoloë hitteresponsproteïen (Hsp70h) kodeer. Die Hsp70 is 'n bewegingsproteïen (MP) wat belangrik is vir die translokasie van die virus deur die plasmodesmata na die naasliggende sel. Breë-spektrum weerstand teen onverwante virusse kan behaal word deur 'n patogeen-afgeleide weerstandstrategie (POR), wat op die uitdrukking van 'n disfunksionele MP wat in plante uitgedruk word, gebaseer is. Die Hsp70hproteïen het twee gebiede. Die N-terminale gebied is In ATPase-gebied en toon hoë homologie met ander ATPase-gebiede van Hsp70h-proteïene van die Closteroviridae, asook die prokariotiese en eukariotiese koninkryke. Gekonserveerde aminosure wat belangrik is vir die posisionering van ATP in die katalitiese domein vir ATP-hidrolise is in die ATPase-gebied gevind. Die C-terminale gebied is variërend en die funksie van die gebied in die Closteroviridae is onbekend. In prokariotiese en eukariotiese Hsp70h-proteïene is die C-terminale gebied belangrik vir proteïenproteïen interaksies. Die nukleotiedvolgorde van die Amerikaanse NY-1-isolaat van GLRaV-3 is al bepaal en POR-strategieë is ook op die kapsiedproteïen, uiteenlopende kapsiedproteïen en die replikasie-proteïen uitgevoer, maar nog nie op 'n disfunksionele vorm van die Hsp70h-geen nie. In hierdie studie is dubbelstring-RNA (dsRNA) van 'n kommersiële wingerd met onbekende virusstatus wat rolblaarsimptome toon, geïsoleer, asook van twee wingerde met 'n bekende virusstatus, een met ligte en een met strawwe simptome. Die GLRaV-3 hsp70h-geen is met hulp van die polimerasekettingreaksie-metode (PKR) vanaf die dsRNA geamplifiseer en die geen se nukleotiedvolgorde is bepaal. Die hsp 70-gene van drie verskillende wingerde het meer as 94% homologie met die NY-1-isolaat getoon. Die gekonserveerde aminosure wat vir ATPase-aktiwiteit belangrik is, was teenwoordig. Die hsp70h-geen van GLRaV-3, wat uit 'n kommersiële wingerd met duidelike rolblaarsimptome in die Stellenbosch-gebied geïsoleer is, is vir verdere navorsing gekies en dit is aan setel-gerigte mutagenese blootgestelom vier mutasies van die geen te bewerkstellig. Die gevolg van hierdie vier mutasies was die verandering van Asn na Asp", Gly na Thr1o, Lys na Glu174 en Asn na Asp197. Die wilde (WT) en veranderde (Mut) vorms van die hsp-gene is in 'n bakteriese uitdrukkingsvektor gekloneer. Uitdrukking van beide die WT- en die Mut-Hspproteïene is behaal, maar die proteïene was in die onoplosbare fraksie geleë. Pogings om die onoplosbare proteïene te isoleer en in 'n aktiewe oplosbare vorm te verkry, was onsuksesvol. Verdere pogings om die proteïene in die oplosbare fraksie van die bakteriese ekspressiesisteem uit te druk, was ook onsuksesvol. As gevolg van die gebrek aan aktiewe proteïen kon biochemiese toetse nie op die ATPaseaktiwiteit van die WT- en Mut-Hsp proteïne gedoen word nie. Die wt- en mut-hsp-gene is ook in In plantekspressievektor gekloneer vir transformasie in tabakplante. Hierdie transformasies was suksesvol en het aanleiding gegee tot 22 kanamisienbestande (Km') en 18 Km' plante vanaf die WT- en Mut-Hspkonstrukte onderskeidelik. Twee plantlyne, M5 en M10, wat met die mut-hsptransgene getransformeer is, het 'n hoë vlak van weerstand teen die infekterende aartappelvirus X getoon in vergelyking met ander plante wat met die virus geïnfekteer is. Daar is dus bewys gelewer dat 'n disfunksionele vorm van die GLRaV-3 Hsp70h weerstand kan bied teen 'n onverwante virus in tabak.

Thesis (MScConsEcol)--Stellenbosch University, 1970. / AFRIKAANSE OPSOMMING: Verskeie aspekte van die biologie van die wingerdfilloksera is bestudeer. Die ondergrondse lewensiklus is in spesiale teelbakke waargeneem en die ontwikkelingstydperk van elke stadium word afsonderlik aangegee. Dit is bepaal dat die ontwikkeling van n eier tot n volwasse radicicolae-wyfie gemiddeld 22 dae duur en dat die ontwikkeling van n eier tot n alata gemiddeld 30 dae duur. Die leeftyd en tydperk van eierlegging van n volwasse wyfie asook die aantal eiers wat geproduseer word, word aangegee. Verskillende gegewens wat in verband met oorwintering verkry is, word bespreek. Die tyd van voorkoms van die verskillende stadia van die radicicolae-siklus in die natuur, is bepaal. Die aktiewe · stadium duur vanaf September tot Mei, die oorwinterende stadium vanaf April tot Oktober en nimfe kom voor vanaf November tot April. Met die gegewens verkry, is die aantal generasies per jaar teoreties bepaal. Filloksera is tot op n diepte van 1.2 m in die grond gevind. Daar is vasgestel dat verskillende grondtipes n invloed op die fillokserabesmetting het. Die besmetting neem af soos die persentasie fyn plus medium sandinhoud van n grond styg. Opnames het getoon dat 95% van die wingerde met Jacquez as onderstok in die Stellenbosch-area besmet is en dat ongeveer 25% van alle wingerde in die Olifantsrivier-besproeiingsgebied besmet is. Met behulp van sekere vangapparate is alatae vir die eerste keer in Suid-Afrika in die natuur versamel. Die leeftyd en eierproduksie van alatae en verskillende aspekte van die voorkoms van die galicolae-siklus word bespreek. Twee metodes is toegepas om morfologiese rasse van filloksera·te onderskei. Met behulp van een van die metodes kon verskille tussen die populasies van die twee geografies verskillende areas wat ondersoek is, aangetoon word. / ENGLISH ABSTRACT: None

Thesis (MScAgric)--Stellenbosch University, 2004. / ENGLISH ABSTRACT: Colonies of Planococcus ficus (Signoret) were reared from three different areas, Hex River Valley, Robertson and Stellenbosch. An insectary colony and a table grape colony from Nietvoorbij experimental farm were also included in the study. A range of concentrations of chlorpyrifos was applied topically to individuals from the different colonies. The Stellenbosch population had the lowest LDso, although it was not significantly different from that of the insectary and Robertson colonies. The Hex River Valley and table grape colonies had a significantly higher LDso than the Robertson, Stellenbosch and insectary colonies, although the relative tolerance was 1.5, which would probably not result in significant control failure in the field. However, this does indicate that there is potential for the development of resistance to chlorpyrifos in the vine mealybug in South Africa. / AFRIKAANSE OPSOMMING: Kolonies van Planococcus ficus (Signoret), is versamel en geteel uit drie verskillende areas, Hex.riviervallei, Robertson en Stellenbosch. 'n Bestaande insektarium kolonie van die Lanbou Navorsings Raad en 'n tafeldruif kolonie vanaf Nietvoorbij proefplaas is ook ingesluit in die studie. 'n Reeks konsentrasies van chlorpyrifos is topikaal aangewend aan individue van die verskillende kolonies. Die Stellenbosch populasie het die laagste LDso getoon alhoewel dit nie betekenisvol verskil het van die LDso van die insektarium - en Robertson kolonies nie. Die Hexriviervallei en tafeldruif kolonies se LDso was betekenisvol hoër as die Robertson, Stellenbosch and insektarium kolonies. Alhoewel die relatiewe weerstand 1.5 was, sal dit waarskynlik nie tot 'n aansienlike beheermislukking in die veld lei nie. Nogtans dui dit op die potensiaal vir moontlike ontwikkeling van weerstand teen chlorpyrifos in die wingerdwitluis.

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Plant-parasitic nematodes, consisting of a wide range of species, can cause severe economic losses in most agricultural food crops. Meloidogyne spp. (root-knot nematodes), Criconemoides xenoplax (ring nematode), Xiphinema index (dagger nematode) and Pratylenchus spp. (lesion nematodes) are some of the economically important plant-parasitic nematodes that pose a threat to viticulture and other perennial crops in South Africa. Worldwide there is ever-increasing pressure on pre-plant synthetic soil fumigants and post-plant nematicides. For sustainable nematode management, it is important to have a holistic approach; taking into consideration cultural, biological and chemical options as part of an integrated management approach. Biofumigation has the potential to fit into such an integrated management system and previous research indicates the positive response on soil-borne diseases, nematodes and weeds. Biofumigation occurs where certain plant species, containing glucosinolates (GSL) in the vacuole of the plant cells, come into contact (after cell maceration), with the enzyme myrosinase (MYR) situated in the cytoplasm of the cell, to form active compounds such as isothiocyanate (ITC). When this green manure is applied to infested soil, the ITC has the potential to have a direct suppressive effect on the soil-borne pathogens and there is also an indirect effect that can be expected after green manure soil amendment, because microbial activity is enhanced in the soil. Brassicas are known to possess GSL and MYR in their cells and thus have the potential to be utilized as biofumigation crops. There are, however, differences in the potential within the Brassicaceae family, based on different types and concentrations of GSL present in the different species. To ensure effective biofumigation it is important to use the correct brassica species and have a good understanding of the factors that have a positive impact on the biofumigation action. Laboratory bioassays were done to determine the potential of different cover crops to suppress Meloidogyne javanica and C. xenoplax, when applied as a green manure. The cover crops used for the bioassays included Oats (Avena sativa cv. Pallinup), White mustard (Sinapis alba cv. Braco), Canola (Brassica napus cv. AV Jade), Caliente 199 (Brassica juncea cv. Caliente 199) and Nemat (Eruca sativa cv. Nemat). The plant material was cut into small pieces and mixed with sterilised soil inoculated with either M. javanica or C. xenoplax. Results from the bioassays showed a significant suppression of M. javanica by the three biofumigation species: White mustard, Caliente 199 and Nemat. These results supported previous research, indicating the nematode suppressing effect due to the biofumigation action of certain brassica crops. Canola did not have the same suppressing impact on the M. javanica and gave comparable results to the control, indicating that Canola is not a good biofumigation crop for M. javanica suppression. In terms of biofumigation effect oats did not differ significantly from the control or the three brassicas: White mustard, Caliente 199 and Nemat. In the bioassays done for C. xenoplax no significant differences were found between the green manure treatments and the control. These results indicate that the different crops tested, including the three well known biofumigation crops, did not suppress the C. xenoplax at the applied biomass concentrations used in the bioassay. Crops can also be classified according to their host status for certain plant parasitic nematodes. Crop host trials were conducted to determine the crop host status of the five different cover crops, to M. javanica and C. xenoplax. The crops were planted in sterilised soil, inoculated with the latter plant-parasitic nematodes and left for 60 days, after which, a root gall index analysis was done for M. javanica and for 85 or 92 days after which C. xenoplax was extracted from the soil. All the crops evaluated had a significantly lower root gall index for M. javanica than the control. Nemat and Oats was classified as poor hosts for M. javanica. A visual inspection of the root systems of all the crops was performed to determine whether M. javanica managed to complete its lifecycle in the different root systems. On all root systems, M. javanica managed to form root galls and produce egg masses, from which (J2) juveniles emerged. This indicates that M. javanica did complete its lifecycle in the different root systems of the crops evaluated and that all the cover crops acted as hosts. The expression of the gall symptoms were, however, less severe on Nemat and Oats, compared to the others. In the C. xenoplax crop host trials, all except the Nemat treatment showed a significant difference, compared to the Tomato treatment, with lower C. xenoplax numbers being present in the other crops. The nematode numbers in the different crops, compared well with the control (only inoculated soil), indicating that the crops did not stimulate the reproduction of C. xenoplax. Canola had the lowest numbers of C. xenoplax present after the growing cycle and Caliente 199 also showed a declining trend. In South Africa, the use of annual cover crops in vineyards is an established soil cultivation practice. In a field study, Oats, White mustard, Canola, Caliente 199 and Nemat were established in a vineyard as cover crops for three growing seasons (2009/10, 2010/11, 2011/12), and evaluated for their biofumigation impact, as well as their host impact on the suppression of certain economically important plant-parasitic nematodes. Two cover crop management practices, namely mechanical incorporation (MC) into the top soil and chemical removal of the cover crop (CC) were applied to the different cover crops. Nematode samples were taken in the work row and in the vine row at different times to determine the nematode status. These periods were April/May, before planting the cover crops, as well as 0, 15, 30 and 60 days after the management practices were performed. The crop biomass, measured as dry matter production (DMP) in tons/ha, differed significantly between the different crops, but also showed substantial increases during the three cover crop growing seasons for most crops. During the three consecutive seasons, Canola (CC) and Caliente 199 (CC) showed a constant reduction in the C. xenoplax population in the vine row based on the 60 day analysis. This trend was also observed for the total plant-parasitic nematode population in the vine row for the three seasons, based on 60 day analysis. The same trend took place during the three-year trial period for all the different sampling periods (0, 15, 30 and 60 days). The results can be attributed to the host status of these crops and not primarily because of the biofumigation effect. Both the Canola (CC) and the Caliente (CC) had a substantial increase in DMP during the three growing seasons that might have played a role in this trend. White mustard (CC and MC) showed a significant increase in the C. xenoplax population in the vine row, over the three year period, based on the 60 day analysis. The same trend was found Nemat (CC) and weeds and nematicide (CC) measured at the same period. A positive result from the Meloidogyne sp. analysis was that there was no significant increase in the Meloidogyne sp. in the vine row during the three growing seasons based on the 60 day analysis. This trend was seen in all the different treatments. The results from this study opens the possibility to apply these cover crops as part of a crop rotation programme without expecting an increase in the Meloidogyne sp. population to occur in the vine row through time. / AFRIKAANSE OPSOMMING: Plantparasitiese nematodes, wat bestaan uit 'n wye verskeidenheid van spesies, kan lei tot ernstige ekonomiese verliese in die meeste landbou gewasse. Meloidogyne spp. (knopwortel nematode), Criconemoides xenoplax (ring nematode), Xiphinema index (dolk nematode) en Pratylenchus spp. (letsel nematode) is van dié belangrikste plantparasitiese nematodes wat 'n bedreiging inhou vir wingerd en ander meerjarige gewasse in Suid-Afrika. Wêreldwyd is daar tans toenemende druk op die uitfasering van voor-plant chemiese grondberoking middels en so ook op nauitplant nematisiede. Vir volhoubare nematode bestuur, is dit belangrik om 'n holistiese benadering te volg, in ag genome kulturele, biologiese en chemiese maatreëls as deel van 'n geïntegreerde benadering. Bioberoking het die potensiaal om deel uit te maak van so 'n geïntegreerde benadering en baie vorige navorsing bevestig hierdie positiewe reaksie, in terme van onderdrukking, wat bioberoking op grond-gedraagde siektes, nematodes en onkruid kan hê. Bioberoking kan beskryf word as die reaksie, wat plaasvind wanneer glukosinolaat (GSL), wat teenwoordig is in die vakuool van die plantselle, in kontak kom met die ensiem mirosinase (MYR), nadat selbreking plaasgevind het en die aktiewe verbinding isothiosianaat (ITC) en ander sekondêre metaboliete gevorm word. Wanneer hierdie groen plantmateriaal in die grond ingewerk word, kan ʼn direkte onderdrukkings effek, as gevolg van die ITC, asook ʼn indirekte onderdrukkings effek as gevolg van die stimulasie van mikrobe aktiwiteit, verwag word. Brassica gewasse is bekend daarvoor dat daar GSL en MYR in die plantselle teenwoordig is en hulle besit dus die potensiaal om ITC te vorm. Daar is egter verskille in hierdie potensiaal binne die Brassicaceae familie, wat gebaseer is op verskillende tipes en konsentrasies GSL. Die keuse van ʼn brassica spesie is dus belangrik, tesame met ʼn verskeidenheid van ander faktore, om optimale bioberoking te verseker. Laboratorium biotoetse is gedoen om die bioberokings effek van verskillende dekgewasse op Meloidogyne javanica en C. xenoplax, wanneer dit aangewend word as groenbemesting, te bevestig. Die dekgewasse wat gebruik is sluit in: Hawer (Avena sativa cv. Pallinup), Wit mosterd (Sinapis alba cv. Braco), Canola (Brassica napus cv. AV Jade), Caliente 199 (Brassica juncea cv. Caliente 199) en Nemat (Eruca sativa cv. Nemat). Die plantmateriaal is fyn opgesny en ingewerk in gesteriliseerde grond wat met onderskeidelik M. javanica en C. xenoplax geïnokuleer is. Resultate van die biotoetse vir M. javanica toon dat die drie gewasse; Wit mosterd, Caliente 199 en Nemat, wat alombekend is vir hul bioberoking potensiaal, ʼn betekenisvolle onderdrukkings op M. javanica tot gevolg gehad het. Hierdie biotoetse ondersteun vorige navorsing, waar effektiewe onderdrukking van sekere Meloidogyne spesies as gevolg van bioberoking verkry is. Die resultate dui ook aan dat Canola nie ʼn goeie opsie is vir effektiewe bioberoking om M. javanica onderdrukking te verkry nie. Die Hawer behandeling het nie betekenisvol van die kontrole of van die ander bioberokings gewasse verskil nie. Daar is geen betekenisvolle verskille verkry tussen die kontrole en die ander gewasse tydens die C. xenoplax biotoetse nie. Die resultate dui aan dat die dekgewasse, insluitende die drie bekende bioberokings gewasse, nie C. xenoplax onderdruk teen die toegediende biomassa konsentrasies nie. Gewasse kan ook geklassifiseer word op grond van hul gasheer status vir sekere nematode. Gasheer toetse is gedoen om die gasheer status van die verskillende dekgewasse vir M. javanica en C. xenoplax te bepaal. Dieselfde vyf verskillende dekgewasse is geplant in grond, wat vooraf onderskeidelik met M. javanica en C. xenoplax geïnokuleer is. Plante is gelos om vir `n spesifieke periode te groei waarna ʼn galindeks evaluasie is gedoen om die gasheer status vir M. javanica te bepaal en ʼn nematode ontleding gedoen is om die gasheer status vir C. xenoplax te bepaal. In die M. javanica gasheer toetse was die galindeks van al die gewasse betekenisvol laer as die kontrole. Nemat kan geklassifiseer word as ʼn swak gasheer vir M. javanica en het betekenisvol minder galle as al die ander gewasse, behalwe die Hawer, waarvan dit nie betekenisvol verskil het nie. Nemat pas dus goed in ʼn dekgewas program waar die doel is om die M. javanica populasie te onderdruk tydens die groei van die gewas. ʼn Visuele inspeksie van die wortelstelsels is ook gedoen ten einde te bepaal of die lewensiklus van M. javanica voltooi is. Wortelgalle en eiersakkies was teenwoordig in die wortels van al die verskillende gewasse en larwes het uit die eiers uitgebroei. Dit dui aan dat M. javanica daarin geslaag het om sy lewenssiklus op al die dekgewasse suksesvol te voltooi. Daar was aansienlik minder eiersakke by Nemat en Hawer; wat hul swak gasheer status bevestig. In die biotoetse vir die gasheerstatus van C. xenoplax het al die gewasse, behalwe Nemat, betekenisvol laer C. xenoplax getalle, in vergelyking met die Tamatie behandeling, tot gevolg gehad. Die nematode getalle was soortgelyk aan die kontrole (slegs geïnokuleerde grond), waar geen gewas in medium geplant is nie, en dui dus aan dat die getalle op die verskillende gewasse nie vermeerder het nie. Die Canola behandeling het die laagste C. xenoplax getalle gehad, gevolg deur Caliente 199. Hierdie gewasse toon dus die meeste potensiaal om aangewend te word in 'n rotasie stelsel of dekgewas program, waar die doel is om die C. xenoplax populasie te onderdruk. In Suid-Afrika is die aanwending van spesifieke eenjarige gewasse, as dekgewasse in wingerde, reeds ʼn standaard praktyk met verskeie voordele. In veldproewe oor ʼn tydperk van drie jaar (2009/10, 2010/11, 2011/12) is Hawer, Wit mosterd, Canola, Caliente 199 en Nemat aangeplant as dekgewasse in ʼn wingerd proefperseel. Die doel van die veldproewe was om die effek van dekgewasse op die plantparasitiese nematodes, wanneer dit aangewend word as bioberokings gewasse, te bepaal. Die gasheer status van die gewasse is ook ondersoek om te bepaal wat die effek sal wees op die nematode getalle. Twee dekgewas bestuurspraktyke is toegepas; meganiese inwerk van die dekgewasse in die bogrond (MC) en chemiese beheer van die dekgewasse (CC) en nematode monsters is op verskillende tye in die werksry en in die wingerdry geneem. Hierdie periodes sluit in April/Mei, voor die vestiging van die dekgewasse en 0, 15, 30 en 60 dae nadat die bestuurspraktyk toegepas is. Die dekgewas se biomassa produksie is, op grond van die droë massa produksie (DMP), in ton/ha gemeet, wat betekenisvol verskil het vir die verskillende dekgewas. Daar het ook `n duidelike toename in DMP plaasgevind oor die drie seisoene vir meeste gewasse. Gedurende die drie jaar periode het die Canola (CC) en Caliente 199 behandelings, gemeet 60 dae na die bestuurspraktyk, ʼn konstante afname getoon in die C. xenoplax in die wingerd ry. Dieselfde tendens het ook voorgekom gedurende hierdie periode in die totale plantparasitiese nematodes teenwoordig in die wingerd ry. Daar is ook ʼn geleidelike afnemende tendens in die C. xenoplax in die wingerd ry, oor die verskillende periodes 0, 15, 30 en 60 dae vir die drie opeenvolgende seisoene, waargeneem. Hierdie resultate kan primêr toegeskryf word aan die gasheer status van die dekgewasse, wat in die gasheer proewe as swak gashere vir C. xenoplax aangetoon is. Nog ʼn faktor wat hier ʼn rol speel is die feit dat beide die Canola (CC) en die Caliente 199 (CC) ʼn toename in DMP van meer as 2 ton, gedurende die drie jaar periode, gehad het; wat op sigself ook ʼn bydraende rol kon speel. Wit mosterd (CC en MC) het oor die drie seisoene ʼn betekenisvolle verhoging in die C. xenoplax populasie tot gevolg gehad, gebaseer op die 60 dae ontleding. Dieselfde tendens is ook opgemerk vir die ander behandelings, onder andere Nemat (CC) en die onkruid en aalwurmdoder (CC) behandeling. ʼn Baie positiewe resultaat na afloop van die drie seisoene is die feit dat daar nie ʼn betekenisvolle verhoging in die Meloidogyne sp. populasie in die wingerdry, op grond van die 60 dae onledings, plaasgevind het nie. Dit was ook die geval vir al die ander behandelings. Hierdie resultate ondersteun die moontlikheid om hierdie bioberokings gewasse deel te maak van ʼn geïntegreerde dekgewas benadering, sonder om in die proses die Meloidogyne sp. in die wingerd ry te verhoog.

Dissertation (PhD (Agric))--University of Stellenbosch, 2005. / ENGLISH ABSTRACT: During the past few years a drastic reduction has been noted in the survival rate of grafted grapevines in nurseries, as well as in young vineyards in the Western Cape Province of South Africa. Circumstantial evidence suggested that Cylindrocarpon spp., which cause black foot disease of grapevine, were associated with this decline. Black foot disease of grapevine is a relatively new, and as yet poorly known disease affecting vines in various countries where grapevines are cultivated. Primary aims of this research have been (1) to conduct nursery surveys in order to determine which fungi are involved in the decline phenomenon, with special reference to the involvement of Cylindrocarpon spp., (2) to identify and characterise the organisms believed to be the causal organisms of black foot disease, and (3) the development of control and/or management strategies to prevent or eradicate Cylindrocarpon infections. Nursery grapevines were sampled at different stages from three commercial nurseries in the Wellington area of the Western Cape Province and were investigated during the 19992000 season by means of destructive sampling. The first samples were taken in September from callused cuttings prior to planting in nurseries. After planting, asymptomatic rooted cuttings were selected from nurseries after 3, 6 and 9 months. Isolation studies clearly demonstrated that different “Cylindrocarpon spp.” infected cuttings from nursery soils. These species rarely occurred in rootstock propagation material prior to planting. At the time of planting, the susceptible basal ends (especially the pith area) of most of the nursery cuttings are partly or even fully exposed. Callus roots also break during the planting process, resulting in small wounds susceptible to infection by soilborne pathogens. The isolation studies revealed that the first infections occurred in the roots, followed by infections of the rootstocks. These infections increased progressively during the course of the growing season. Substantial variation in cultural and morphological characters was observed among the Cylindrocarpon isolates obtained from the nursery survey, as well as from isolations that were made from diseased grapevines. Morphological and phylogenetic studies were conducted to identify these “Cylindrocarpon spp.” and to establish their association with black foot disease. Sequences of the partial nuclear large subunit ribosomal DNA (LSU rDNA), internal transcribed spacers 1 and 2 of the rDNA including the 5.8S rDNA gene (ITS), and partial β-tubulin gene introns and exons were used for phylogenetic inference. Phylogenetic analyses confirmed the diversity observed among the isolates and four Cylindrocarpon-like species were identified. One of these species was initially identified as Cylindrocarpon destructans. However, further research revealed C. destructans to represent a species complex. Grapevine isolates of “C. destructans” proved to be identical to the ex-type strain of Cylindrocarpon liriodendri, which also produced a teleomorph, Neonectria liriodendri in culture. A second species was newly described in this study as Cylindrocarpon macrodidymum (Neonectria macrodidyma). The two remaining Cylindrocarpon-like species were placed in a new genus, Campylocarpon. The two species were named Campylocarpon fasciculare and Campylocarpon pseudofasciculare. Pathogenicity studies confirmed that all four species were able to reduce root and shoot mass significantly. Knowledge obtained pertaining to the disease cycle of black foot disease suggest that suitable management strategies should focus on prevention of primary infection in nurseries. However, at present, no fungicides are registered for control of this disease in South African vineyards or nurseries. Thirteen fungicides were screened in vitro for mycelial inhibition of these pathogens. Prochloraz manganese chloride, benomyl, flusilazole and imazalil were the most effective fungicides tested, and were subsequently included in semi-commercial field trials. Basal ends of grafted cuttings were dipped (1 min) in various chemical and biological treatments prior to planting in open-rooted nurseries. Black foot pathogens were not isolated from grafted cuttings prior to planting in nurseries. Additional treatments involved soil amendments with Trichoderma formulations and hot water treatment (50°C for 30 min) of dormant nursery grapevines. Field trials were evaluated after a growing season of eight months. The incidence of black foot pathogens was not significantly and/or consistently reduced by the majority of chemical or biological treatments. However, these pathogens were not isolated from uprooted plants that were subjected to hot water treatment. It is therefore recommended that hot water treatment of dormant nursery plants be included in an integrated strategy for the proactive management of black foot disease in grapevine nurseries. / AFRIKAANSE OPSOMMING: Gedurende die afgelope paar jaar is ‘n drastiese afname waargeneem in die sukses van geënte wingerdplante in kwekerye, sowel as jong wingerde van die Wes-Kaap. Omstandigheidsgetuienis dui daarop dat Cylindrocarpon spp., wat die wingerdsiekte swartvoet veroorsaak, geassosieer word met hierdie agteruitgang. Swartvoet is ‘n relatiewe nuwe siekte waarvan daar baie min inligting bekend is, alhoewel dit voorkom in verskeie lande waar wingerd verbou word. Die primêre doel van navorsing was (1) om opnames in wingerdkwekerye uit voer om te bepaal watter swamme betrokke is by die verskynsel van agteruitgang, met spesiale verwysing na die betrokkenheid van Cylindrocarpon spp., (2) om die organismes te identifiseer en te karakteriseer wat daarvan verdink word dat hulle die siekte swartvoet veroorsaak, en (3) om beheer en/of bestuurspraktyke te ontwikkel om Cylindrocarpon infeksies te voorkom of uit te wis. Kwekeryplantjies in drie kommersiële kwekerye in die Wellington omgewing van die Wes-Kaap is gedurende verskillende tye gedurende die groeiseisoen gemonitor. Die opnames het plaasgevind gedurende die 19992000 seisoen deur middel van destruktiewe monsterneming. Die eerste monsters is geneem in September nadat die stokkies geënt en gekallus is en voordat dit in die kwekery geplant is. Na plant is asimptomatiese, gewortelde plante vanuit die kwekerye na 3, 6 en 9 maande uitgehaal. Isolasiestudies dui duidelik daarop dat verskillende “Cylindrocarpon spp.” plante vanuit die kwekerygrond geïnfekteer het. Hierdie spesies het selde voorgekom in onderstok-voortplantingsmateriaal voor plant. Tydens plant is die vatbare basale gedeelte, veral die pit, van die meeste geënte stokkies gedeeltelik of selfs volledig blootgestel. Kalluswortels breek ook tydens plant wat wonde laat vir infeksie deur grondgedraagde siektes. Die isolasiestudies dui ook daarop dat die eerste infeksies in die wortels plaasgevind het, gevolg deur infeksies van die onderstokke. Hierdie infeksies het toenemend voorgekom gedurende die verloop van die groeiseisoen. Substansiële variasie in kultuur- en morfologiese eienskappe is waargeneem in die Cylindrocarpon isolate wat tydens die kwekeryopnames versamel is, sowel as van isolasies wat gemaak is uit siek plante. Morfologiese en filogenetiese studies is uitgevoer om hierdie “Cylindrocarpon spp.” te identifiseer en hul betrokkenheid by die siekte swartvoet uit te klaar. Gedeeltelike DNS volgordes van die groot ribosomale subeenheid (“LSU rDNA”), interne getranskribeerde spasiëerderarea (“ITS1, “ITS2”), insluitend die 5.8S rRNS geen, en gedeeltelike β-tubilien geen introns and eksons is gebruik vir filogenetiese analise. Filogenetiese analises het die diversiteit wat waargeneem is tussen die verskillende isolate bevestig deurdat vier Cylindrocarpon-agtige spesies geïdentifiseer is. Een van hierdie spesies is aanvanklik geïdentifiseer as Cylindrocarpon destructans. Verdere navorsing het egter daarop gedui dat C. destructans ‘n spesie-kompleks verteenwoordig. “C. destructans” afkomstig van wingerd blyk identies te wees aan die ex-tipe isolaat van Cylindrocarpon liriodendri, wat ook ’n teleomorf, Neonectria liriodendri in kultuur vorm. ’n Tweede spesie is nuut beskryf in hierdie studie as Cylindrocarpon macrodidymum (Neonectria macrodidyma). Die twee oorblywende Cylindrocarpon-agtige spesies is geplaas in ‘n nuwe genus, Campylocarpon. Die twee spesies staan bekend as Campylocarpon fasciculare en Campylocarpon pseudofasciculare. Patogenisiteitstudies het bevestig dat al vier spesies die vermoë het om wortel- en lootmassa van wingerdplant drasties te verlaag. Kennis wat opgedoen is rakende die lewensiklus van swartvoet dui daarop dat bestuurspraktyke daarop moet fokus om primêre infeksies in wingerdkwekerye te voorkom. Op die oomblik is daar egter geen fungisiedes geregistreer vir die beheer van die siekte in Suid- Afrikaanse wingerde of kwekerye nie. Dertien fungisiedes is in vitro geëvalueer om te bepaal of dit miseliumgroei van hierdie swamme kan inhibeer. Prochloraz mangaan chloried, benomyl, flusilasool en imazalil was die effektiefste fungisiedes wat ondersoek is, en is gevolglik ingesluit in semi-kommersiële veldproewe. Die basale gedeelte van geënte stokkies is gedoop (1 min) in verskeie chemies en biologiese behandelings voordat dit geplant is in die kwekerye. Patogene wat geassosieer word met swartvoet is nie vanuit geënte stokkies geïsoleer voordat dit in die kwekerye geplant is nie. Addisionele behandelings het bestaan uit grondtoevoegings met Trichoderma formulasies, sowel as warmwaterbehandeling (50°C vir 30 min) van dormante kwekeryplante. Die veldproewe is geëvalueer na ‘n groeiseisoen van 8 maande. Die voorkoms van swartvoet patogene is nie betekenisvol/konstant verlaag deur die meeste chemies en biologiese behandelings nie. Hierdie patogene is egter nie vanuit plante geïsoleer wat na uithaal aan warmwaterbehandeling blootgestel is nie. Dit word dus aanbeveel dat warmwaterbehandeling van dormante kwekeryplante deel word van ‘n geïntegreerde strategie vir die pro-aktiewe beheer van swartvoet in wingerdkwekerye.

No description available.

Grapevine powdery mildew is caused by Erysiphe necator, an Ascomycete fungus and an obligate biotroph restricted to growth on its grapevine host. Biotrophic pathogens form a stable association with host cells without directly causing cell death, and take up nutrients from, in the case of powdery mildew ( PM ), host epidermal cells ( Rumbolz et al., 2000 ). As the fungus grows, its increasing biomass becomes a strong nutrient sink capable of altering assimilate flow and storage in the host. To identify host genes that may mediate nutrient delivery to powdery mildew infected tissues and therefore may contribute to disease susceptibility, a candidate gene approach using degenerate and RT - PCR, and a nontargeted approach using microarray analysis was instigated. Once identified, " susceptibility genes " could be targeted for manipulation to provide alternative resistance strategies based on reduced susceptibility in the future. In addition to genes encoding pathogenesis and stress related proteins, microarray analysis revealed that transcript levels of a putative metal transporter and a cell wall structural protein were elevated in infected berry skin, while aquaporin water channels and genes associated with photosynthesis were generally repressed. Degenerate PCR was used to isolated new cell wall invertase, monosaccharide and amino acid transporter genes and initial RT - PCR revealed that expression of genes involved in sugar mobilisation were the most significantly modulated by powdery mildew infection. Previously unreported hexose transporters ( HTs ), ( VvHT3, VvHT4 and VvHT5 ) and a cwINV ( VvcwINV ) had been isolated from cDNA prepared from powdery mildew infected grapevine leaves. Full length clones of grapevine HTs and cwINV were obtained by RACE PCR. Heterologous expression of the three new HTs in yeast confirmed that VvHT4 and VvHT5 mediated glucose uptake, while VvHT3 did not function in the yeast system. However, transient expression of a translational fusion of the VvHT3 protein with green florescence protein in onion epidermal cells indicated that it is targeted to the plasma membrane of plant cells. Quantitative RT - PCR analysis of these new genes, together with previously reported grapevine HTs and cytoplasmic and vacuolar invertases, indicated that expression of VvcwINV and VvHT5, were significantly up - regulated by PM infection, while a vacuolar invertase was strongly down - regulated by PM infection. Invertase activity assays were in agreement with these findings, showing elevated sucrolytic activity in insoluble fractions and reduced sucrolytic activity in soluble fractions. These results suggest that apoplasmic phloem unloading of sucrose in the infected leaf is elevated and that VvHT5 is induced to recover the additional hexoses from the apoplasm. Basic localisation studies indicated that VvHT5 and VvcwINV are not induced specifically in powdery mildew infected leaf regions, but are induced in a more diffuse distribution within infected leaves. To determine if induction of VvHT5 and VvcwINV is specific to PM infection or if other stimuli may also mediate these responses, leaves were inoculated with downy mildew or stressed by wounding. Transcript levels of VvHT5 and VvcwINV were elevated by wounding and downy mildew infection, suggesting that the induction of these genes may be part of a general stress response. To explore the signalling pathways that may underlie these responses, leaves were treated with the plant growth regulators ethylene, jasmonate and abscisic acid. Exogenous application of ethylene and methyl jasmonate only marginally affected the expression of the genes studied, however foliar application of abscisic acid ( ABA ) induced gene expression changes similar to those observed in response to powdery mildew infection and wounding. Promoter sequences of VvHT3, VvHT4, VvHT5 and VvcwINV were isolated and analysed for the presence of regulatory elements. Compared with the promoters of VvHTs that were not induced by pathogen infection or wounding, the VvHT5 and VvcwINV promoters contained numerous motifs associated with induction by ABA including ABRE, Myc and Myb binding elements. The path of sugar loading into the mesocarp of grape berries during ripening is still poorly understood and few molecular components associated with this process have been described. Quantitative RT - PCR was used to monitor the expression of five HTs and VvcwINV during Cabernet sauvignon and Shiraz berry development and ripening. Of the three new HTs reported here, the expression of VvHT3 is most consistent with a potential role in sugar loading, while VvHT5 is induced late in this process. VvcwINV transcript levels were high pre - ripening and also during the later stages of ripening, therefore based on this expression pattern, a role for this enzyme during ripening is not clearly evident. These results are discussed in terms of an apoplasmic step in phloem unloading in ripening grape berries. This study has provided new insights into the molecular and biochemical processes associated with the formation of carbohydrate sink metabolism in response to stress stimuli, and sugar delivery to grape berries during ripening. ABA - dependant pathways may mediate the stress - associated induction of VvcwINV and VvHT5, presumably to recruit additional carbohydrates to the affected organ to energise repair and defence responses. At this stage it is unknown if this response is beneficial to pathogen nutrition, however potentially, modification of genes associated with carbohydrate sink metabolism could provide an alternative way to engineer resistance to this pathogen. / Thesis (Ph.D.)--School of Agriculture, Food and Wine, 2006.

There is currently little knowledge about the dynamics of invertebrates in Australian viticultural ecosystems. This study was conducted in Coonawarra vineyards over three seasons (years) and has focused on identifying natural enemies, their seasonal phenology, multiple species interactions, and potential for the suppression of the pest lepidopteran Epiphyas postvittana (Tortricidae). The work presented in this thesis shows that endemic natural enemies have far greater potential to control E. postvittana than has been realised. An initial survey identified a diverse and abundant range of potential natural enemies. Of these, the species most likely to attack E. postvittana include a predatory mite Anystis baccarum and a number of hymenopteran parasitoids. The most abundant parasitoid in the vineyards was a braconid, Dolichogenidea tasmanica. Understanding the characteristic behaviour of parasitoids in response to host density can help to gauge their potential for pest suppression. The results of large-scale field experiments showed that the response of D. tasmanica to the density of E. postvittana was inversely density-dependent, and that parasitism was consistently higher in Cabernet Sauvignon compared with Chardonnay varieties. Despite the fact that interactions among multiple species of natural enemies can increase or decrease pest suppression, particularly when they share a common prey/host, few multispecies interactions have been investigated. Laboratory studies identified a novel interaction between the predatory mite A. baccarum an abundant predator in the vine canopy, the parasitoid D. tasmanica and host E. postvittana larvae. Although A. baccarum readily ate E. postvittana eggs and free roaming larvae, they could not access larva in their silk leaf rolls. However, the addition of D. tasmanica significantly increased predation of E. postvittana larvae, by altering the behaviour of host larvae and increasing their vulnerability to the mite. Experiments conducted at a landscape level in the Coonawarra showed that D. tasmanica was also present in habitat other than vineyards including native vegetation. However, it was not present in highly disturbed habitats. Although the exact mechanism for this remains unknown, results indicate that viticultural practices and resources in the surrounding landscape can influence the presence of parasitoids. Together, the findings presented in this thesis make a significant contribution towards developing sustainable pest management in Australian viticulture. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1320930 / Thesis (Ph.D.) -- University of Adelaide, School of Earth and Environmental Sciences, 2008

There is currently little knowledge about the dynamics of invertebrates in Australian viticultural ecosystems. This study was conducted in Coonawarra vineyards over three seasons (years) and has focused on identifying natural enemies, their seasonal phenology, multiple species interactions, and potential for the suppression of the pest lepidopteran Epiphyas postvittana (Tortricidae). The work presented in this thesis shows that endemic natural enemies have far greater potential to control E. postvittana than has been realised. An initial survey identified a diverse and abundant range of potential natural enemies. Of these, the species most likely to attack E. postvittana include a predatory mite Anystis baccarum and a number of hymenopteran parasitoids. The most abundant parasitoid in the vineyards was a braconid, Dolichogenidea tasmanica. Understanding the characteristic behaviour of parasitoids in response to host density can help to gauge their potential for pest suppression. The results of large-scale field experiments showed that the response of D. tasmanica to the density of E. postvittana was inversely density-dependent, and that parasitism was consistently higher in Cabernet Sauvignon compared with Chardonnay varieties. Despite the fact that interactions among multiple species of natural enemies can increase or decrease pest suppression, particularly when they share a common prey/host, few multispecies interactions have been investigated. Laboratory studies identified a novel interaction between the predatory mite A. baccarum an abundant predator in the vine canopy, the parasitoid D. tasmanica and host E. postvittana larvae. Although A. baccarum readily ate E. postvittana eggs and free roaming larvae, they could not access larva in their silk leaf rolls. However, the addition of D. tasmanica significantly increased predation of E. postvittana larvae, by altering the behaviour of host larvae and increasing their vulnerability to the mite. Experiments conducted at a landscape level in the Coonawarra showed that D. tasmanica was also present in habitat other than vineyards including native vegetation. However, it was not present in highly disturbed habitats. Although the exact mechanism for this remains unknown, results indicate that viticultural practices and resources in the surrounding landscape can influence the presence of parasitoids. Together, the findings presented in this thesis make a significant contribution towards developing sustainable pest management in Australian viticulture. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1320930 / Thesis (Ph.D.) -- University of Adelaide, School of Earth and Environmental Sciences, 2008