M.S.

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Ethel M. Doidge's thesis is placed on the UIR in 2013 to celebrate 140 years of university education in South Africa. She was the first women to receive a D.Sc. from Unisa'a predecessor, the University of the Cape of Good Hope in 1914 / This thesis researched a mango disease which caused considerable loss to mango growers in South Africa at the beginning of the twentieth century, particularly around Barberton, Warm baths and the coastal region of Natal. The disease caused dark angular spots on the leaves. This did not noticeably affect the general health of the tree, but served as a source of infection for the fruit. The infected fruit is detached from the tree by the slightest air movement and falls rotting to the ground. The infection is carried by wind and rain. Several spraying experiments were carried out at Barberton in an orchard placed at the author's disposal by Messrs Winter Brothers during an exceptionally dry season, which showed that spraying with Bordeaux mixture, iron sulphide or Hyco/ was useless in checking the disease. The disease had not been described in the literature before. According to the author, the cause of the disease was a flagellate bacillus Bacillus mangiferae. It invades the parenchyma, wedging apart and killing the cells and causing gummosis, but it does not touch the lignified tissues. The organism is described, and was tested in detail in laboratory experiments and a resume of its salient characters is given in the thesis / Agriculture and  Animal Health / D.Sc. (Botany)

Thesis (MSc)--Stellenbosch University, 1968. / Please refer to full text for abstract

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)--Stellenbosch University, 2001. / ENGLISH ABSTRACT: Bacterial canker of stone fruit, caused by Pseudomonas syringae pv. syringae, is one of the most destructive crop diseases in South Africa. Chemical control has failed completely and effective long-term management strategies will have to rely on the breeding of resistant host trees. To assist in such breeding programmes, investigations into the molecular basis of the interaction between P. s. pv. syringae and stone fruit trees have been undertaken in collaboration with the ARC-Fruit, Wine and Vine Research Institute in Stellenbosch. The aim of this dissertation was to clone and identify genes that are involved in interaction between the bacterial canker pathogen and stone fruit trees. In the first part of the study, the harpin encoding gene of a local strain of the pathogen, P. s. pv. syringae NV, was amplified in a polymerase chain reaction (PCR) strategy with primers based on the hrpAZB sequences of the bean pathogen, P. s. pv. syringae 61. Sequencing of this hrpZpssNvgene revealed a high degree of homology (96%) between the harpin encoding genes and harpin proteins of the two strains. The hrpZpssNvgene was subsequently cloned into the pMAL-c2 expression vector and expressed in Escherichia co/i. This system was used for the production of purified, biologically active, recombinant HrpZpSSNV protein. In the second part of the study, differential display (DD) technology was used to identify genes that are induced in stone fruit trees in response to P. s. pv. syringae and/or its harpin elicitor. For this purpose, actively growing shoots of two Prunus sa/icina cultivars, the moderately resistant cv. 'Laetitia' and the highly susceptible cv. 'Songold' were treated with recombinant harpinpssNvprotein or live P. s. pv. syringae NV bacteria. An untreated control and wounding control was included in the experiment. Total RNA was isolated for comparative mRNA analysis 24 hours after treatment. DD profiles were generated with fifteen primer combinations. Eight candidate bands were re-amplified, cloned and sequenced. Reverse transcription PCR was employed to verify the expression patterns of the cloned bands in the original RNA sample set. Two bands, DDc and DD4 were shown to be differentially expressed between treatments and/or cultivars, while no differences in the expression levels of the remaining six bands (DDa, DDe, DD3, DD5, DD6 and DD7) were observed. BLAST similarity searches yielded significant matches for DDe, DD4 and DD7 with plant defense-related genes. / AFRIKAANSE OPSOMMING: Bakteriese kanker van steenvrugte, wat deur Pseudomonas syringae pv. syringae veroorsaak word, is een van die mees verwoestende siektes van landbougewasse in Suid-Afrika. Chemiese beheermaatreëls het geheel en al misluk en effektiewe langtermyn beheerstrategieë sal op die teling van weerstandbiedende gasheerbome moet staatmaak. Ondersoeke na die molekulêre basis van die interaksie tussen P. s. pv. syringae en steenvrugbome is in samewerking met die LNR-Vrugte-, Wyn- en Wingerdnavorsingsinstituut in Stellenbosch van stapel gestuur om tot sulke telingsprogramme by te dra. Die doelwit van hierdie proefskrif was om gene wat betrokke is in die interaksie tussen die bakteriese kanker patogeen en steenvrugbome te kloneer en te identifiseer. In die eerste gedeelte van die studie is die harpien-koderende geen van 'n plaaslike ras van die patogeen, P. s. pv. syringae NV, geamplifiseer in 'n polimerase kettingreaksie (PKR)-strategie met peilers wat op die hrpAZB-geenopeenvolgings van die boontjiepatogeen, P. s. pv. syringae 61, gebaseer is. Volgordebepaling van hierdie hrpZpssNv-geen het 'n hoë vlak van homologie (96%) tussen die harpien-koderende gene en harpien proteïene van die twee rasse getoon. Die hrpZpssNv-geen is vervolgens in die uitdrukkingsvektor pMAL-c2 gekloneer en uitgedruk in Escherichia coli. Hierdie sisteem is vir die produksie van suiwer, biologies-aktiewe, rekombinante HrpZpssNv-proteingebruik. In die tweede gedeelte van die studie is die differensiaalvertoon (DD) tegniek gebruik om gene te identifiseer wat deur P. s. pv. syringae en/of sy harpien elisitar in steenvrugbome geïnduseer word. Vir hierdie doel is aktief-groeiende lote van twee Prunus sa/icina kultivars, die matig weerstandbiedende kv. 'Laetitia' en die hoogs vatbare kv. 'Songold', met rekombinante harpienpssNvproteïen of lewende P. s. pv. syringae NV bakterieë behandel. 'n Onbehandelde- en verwondingskontrole is in die eksperiment ingesluit. Totale RNA is 24 uur na behandeling vir vergelykende mRNA-analise geïsoleer. DD-profiele is met vyftien peilerkombinasies gegenereer. Agt kandidaatbande is geheramplifiseer en gekloneer, waarna hul DNA-opeenvolgings bepaal is. Trutranskriptase-PKR is gebruik om die ekspressiepatrone van die gekloneerde bande in die oorspronklike RNA monsters na te gaan. Daar is vasgestel dat twee van die bande, DDc en DD4, differensieel tussen kultivars en/of behandelings uitgedruk is, terwyl geen verskille in die ekspressievlakke van die oorblywende ses bande (DDa, DOe, 003, DOS, 006 en DO7) waargeneem is nie. BLAST-soektogte het betekenisvolle ooreenkomste vir DDe, DD4 en DD7 met plant weerstandsgeassosieerde gene opgelewer.

Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: The codling moth, Cydia pomonella (Lepidoptera: Tortricidae), is the key pest of apples and pears worldwide. The withdrawal of certain fundamental chemicals from codling moth management spray programmes, due to concerns about human, environmental and ecosystem health, has resulted in the search for softer, more environmentally friendly, and safer control measures. Entomopathogenic nematodes (EPNs), naturally occur in the soil, and actively search for hosts. The interest in using EPNs from the families Heterorhabditidae and Steinernematidae as a control measure was sparked in 1953, when an EPN was discovered in an insect. The aim is to incorporate EPNs in an integrated pest management (IPM) programme, to ensure minimal residue and eventually residue-free pome fruit production in South Africa. In order to ensure EPN success, both the environmental and technical factors influencing their efficacy, were investigated in this study. The biocontrol potential of three imported EPN isolates, being Steinernema feltiae and two isolates of Heterorhabditis bacteriophora (Hb1, Hb2), as well as a local isolate, Steinernema yirgalemense, were evaluated for the control of the codling moth under local conditions. All concentrations of S. yirgalemense, applied by immersion in a suspension of nematodes, gave > 98% control. The two formulated isolates of H. bacteriophora, Hb1-f and Hb2-f, gave < 30% control. When using the same nematode isolates, produced in vivo, S. yirgalemense still resulted in a higher codling moth control of > 90%, compared to 54% and 31% control of the H. bacteriophora Hb1 and Hb2 isolates, respectively. In follow up field trials, S. feltiae resulted in ≥ 80% control, and was thus more effective than both S. yirgalemense and the H. bacteriophora (Hb1) isolates, with 66% and 24%, and 24% and 9% control, for two separate trials, respectively. To validate the data obtained from the field trials, subsequent laboratory bioassays were conducted evaluating temperature regimes, following the same cycle as under natural conditions, with a constant humidity of 100%. Steinernema feltiae proved to be most effective, causing > 90% mortality, followed by S. yirgalemense, with 78% mortality. The two H. bacteriophora isolates (Hb1, Hb2) under the above-mentioned laboratory conditions, resulted in 73% and 59% control, respectively. Humidity thus seems to be the most important factor affecting EPN efficacy during above-ground applications. Steinernema feltiae proved to be a better candidate than S. yirgalemense for the control of the codling moth. The efficacy of different EPN isolates in controlling diapausing codling moth larvae at different temperatures was also evaluated, under local conditions, using spray application. Steinernema feltiae and two isolates of H. bacteriophora Hb1 and Hb2, including two local isolates, S. yirgalemense and Steinernema jeffreyense, were evaluated. The use of S. jeffreyense resulted in the most effective control, with 67% mortality, followed by H. bacteriophora (Hb1) with 42%, and then by S. yirgalemense with 41%. Laboratory bioassays simulating field conditions revealed that S. feltiae was most virulent to codling moth larvae, with 67% mortality by infection, followed by S. yirgalemense with 58%, the H. bacteriophora strain Hb1 with 48%, and the Hb2 strain with 24%. A comparison of the infection and penetration rate of two isolates of H. bacteriophora (Hb1, Hb2), S. feltiae and S. yirgalemense, which was carried out in multiwell plates at 14°C and 25°C, respectively, confirms the dramatic effect of temperature on EPN efficacy. At 14°C, all treatments with EPN species resulted in slower codling moth mortality than they did at 25°C, as after 48 h, < 15% mortality was recorded for all species, whereas at the warmer temperature, > 98% mortality was recorded for all species. After the exposure of washed, cool-treated larvae to 25°C for 24 h, the application of both S. feltiae and S. yirgalemense resulted in 100% mortality, whereas the application of the two H. bacteriophora isolates, Hb1 and Hb2, resulted in 68% and 54% control, respectively, over the same time period. At 14°C, S. feltiae had the highest average penetration rate of 20 IJs/insect, followed by S. yirgalemense with 14 IJs/insect, whereas S. yirgalemense had the highest penetration rate at 25°C, with 39 IJs/insect, followed by S. feltiae, with 9 IJs/insect. The two H. bacteriophora isolates had higher average penetration rates at the higher temperature. This study has highlighted the biocontrol potential of S. jeffreyense, as well as showing that S. feltiae is a cold-active nematode, whereas the other three EPN isolates prefer warmer temperatures. Stacked wooden fruit bins are regarded as preferred overwintering sites for codling moth diapausing larvae. Control strategies against the codling moth in South Africa have been hampered by the reinfestation of orchards by nearby stacked infested fruit bins or by the movement of bins between orchards. Worldwide, wooden fruit bins are systematically being replaced with plastic bins, which, in South Africa, will only be phased out over a few years. The objective of this study was to evaluate the potential of H. bacteriophora, S. feltiae, and S. yirgalemense, to disinfest miniature wooden fruit bins under controlled conditions in the laboratory. After dipping minibins in a suspension of 25 IJs/ml of all three EPN species, under optimum conditions of temperature and humidity, the highest percentage of control was obtained using S. feltiae (75%) followed by S. yirgalemense (57%), and then by H. bacteriophora (Hb1) (27%). The addition of adjuvants significantly increased (p < 0.001) S. feltiae infectivity to > 95%, whereas it did not result in a significant increase in H. bacteriophora or S. yirgalemense infectivity. The results indicated that H. bacteriophora would not be a suitable candidate to use for the control of the codling moth larvae in wooden fruit bins. The current preferred candidate for control would be S. feltiae, whose efficacy could be increased by means of the addition of an adjuvant. During winter, when the whole codling moth population are larvae and in diapause, no control measures are applied in orchards. This study has shown that EPNs can be sprayed in orchards to lower the codling moth cohort emerging after winter, as well as be included in an IPM programme. EPNs can act as a second line of defence, through supplementary control, and ensure effective control of the codling moth larvae which survived chemical spray applications, to safeguard against resistant codling moth populations in the next season. / AFRIKAANSE OPSOMMING: Kodlingmot, Cydia pomonella (Lepidoptera: Tortricidae), is ‘n belangrike plaag van appels en pere wêreldwyd. Die onttrekking van sekere fundamentele chemikalieë vanuit die kodlingmot beheerprogram weens die kommer oor menslike, omgewings en ekosisteemgesondheid, het gelei tot die soektog na sagter, meer omgewingsvriendelike en veiliger beheermaatreëls. Entomopatogeniese nematodes (EPNs) kom natuurlik in die grond voor en soek aktief na gashere. Die belangstelling in die gebruik van EPNs van die families Heterorhabditidae en Steinernematidae as 'n beheermaatreël is te danke aan die ontdekking van 'n EPN in ‘n insek in 1953. Die doel is om EPNs in 'n geïntegreerde plaagbeheerprogram (GPB) te inkorporeer om sodoende minimale residue te verseker en uiteindelik residu vrye produksie van kernvrugte in Suid-Afrika. Ten einde die sukses van EPNs te verseker, is beide die omgewings- en tegniese faktore wat hul doeltreffendheid beïnvloed in die studie ondersoek. Die biologiese beheer potensiaal van drie ingevoerde EPN isolate, Steinernema feltiae en twee Heterorhabditis bacteriophora (Hb1, Hb2) isolate, sowel as 'n plaaslike isolaat, Steinernema yirgalemense, is vir die beheer van kodlingmot onder plaaslike toestande geëvalueer. Alle konsentrasies van S. yirgalemense, wat deur indompeling in ‘n suspensie van nematodes toegedien is, het > 98% beheer tot gevolg gehad. Die twee geformuleerde isolate van H. bacteriophora, Hb1-f en Hb2-f, het < 30% beheer gegee. Met die gebruik van dieselfde nematode isolate, wat in vivo geproduseer is, het S. yirgalemense nog steeds 'n hoër kodlingmot beheer van > 90% opgelewer, in vergelyking met die 54% en 31% beheer van die H. bacteriophora Hb1 en Hb2 isolate, onderskeidelik. Steinernema feltiae het in opvolg veldproewe ≥ 80% beheer tot gevolg gehad en was dus meer effektief as beide S. yirgalemense en die H. bacteriophora (Hb1) isolate, met 66% en 24% en 24% en 9% beheer onderskeidelik in twee afsonderlike veldproewe. Om die resultate van die veldproewe te bevestig, is daaropvolgende laboratorium biotoetse uitgevoer en temperatuur regimes is geëvalueer deur die selfde siklus as onder natuurlike toestande te volg, met 'n konstante humiditeit van 100%. Die studie het bewys dat S. feltiae die mees doeltreffende isolate was met > 90% mortaliteit, S. yirgalemense het gevolg met 78% mortaliteit. Die twee H. bacteriophora isolate (Hb1, Hb2) het onderskeidelik onder bogenoemde laboratorium toestande 73% en 59% beheer tot gevolg gehad. Humiditeit blyk dus die belangrikste faktor te wees wat EPN se doeltreffendheid tydens bogrondse toediening affekteer. Die studie het bewys dat S. feltiae 'n beter kandidaat as S. yirgalemense vir die beheer van kodlingmot is. Die doeltreffendheid van verskillende EPN isolate vir die beheer van diapause kodlingmot larwes sowel as EPN se aktiwiteit by verskillende temperature is ook onder plaaslike toestande, deur bogrondse bespuitings, geëvalueer. Steinernema feltiae en twee isolate van H. bacteriophora (Hb1, Hb2), S. yirgalemense en ‘n ander plaaslike isolaat, Steinernema jeffreyense, is geëvalueer. Die gebruik van S. jeffreyense, het tot die mees effektiewe beheer gelei, met 67% mortaliteit, gevolg deur H. bacteriophora (Hb1) met 42%, en dan S. yirgalemense met 41%. Laboratorium biotoetse wat veldtoestande simuleer, het bewys dat S. feltiae die mees doeltreffend teen kodlingmot larwes is, met 67% mortaliteit tydens infeksie, gevolg deur S. yirgalemense met 58%, die H. bacteriophora Hb1 isolaat met 48%, en die Hb2 isolaat met 24%. 'n Vergelyking van die infeksie- en penetrasie tempo van twee isolate van H. bacteriophora (Hb1, Hb2), S. feltiae en S. yirgalemense wat in 12-put plate teen 14°C en 25°C uitgevoer is, het die dramatiese effek van temperatuur op EPN doeltreffendheid bevestig. By 14°C het alle EPN spesies behandelings stadiger kodlingmot mortaliteit as by 25°C na 48h tot gevolg gehad. ‘n Mortaliteit van < 15% is vir alle spesies aangeteken terwyl by die warmer temperature is > 98% mortaliteit vir alle spesies aangeteken. Na die blootstelling van afgespoelde, koel behandelde larwes aan 25°C vir 24 uur, het die toediening van beide S. feltiae en S. yirgalemense, 100% mortaliteit van larwes tot gevolg gehad terwyl die toediening van die twee H. bacteriophora isolate, Hb1 en Hb2, onderskeidelik 68% en 54% beheer tot gevolg gehad, oor dieselfde tydperk. By 14°C, het S. feltiae die hoogste gemiddelde penetrasie tempo van 20 ILs/ larwe, gevolg deur S. yirgalemense met 14 ILs/ larwe tot gevolg gehad, terwyl S. yirgalemense die hoogste penetrasie tempo getoon het by 25°C met 39 ILs/ insek, gevolg deur S. feltiae met 9 ILs/ insek. Die twee H. bacteriophora isolate (Hb1 en Hb2) het ook hoër gemiddelde penetrasie tempo by die hoër temperatuur getoon. Hierdie studie het die biobeheer potensiaal van S. jeffreyense beklemtoon, asook weereens bevestig dat S. feltiae ‘n koue-aktiewe nematode is, terwyl die ander drie EPN isolate warmer temperature verkies. Hout vrugtekratstapels, word beskou as ‘n ideale oorwintering skuiling vir kodlingmot diapause larwes. In Suid-Afrika word beheerstrategieë teen kodlingmot in die wiele gery deur die herbesmetting van boorde deur nabygeleë besmette hout vrugtekratte of deur die beweging van kratte tussen boorde. Hout vrugtekratte word wêreldwyd stelselmatig vervang met plastiek kratte. Dit sal egter eers oor ‘n aantal jare in Suid-Afrika uitgefaseer word. Die doel van hierdie studie was om die potensiaal van H. bacteriophora, S. feltiae, en S. yirgalemense te evalueer deur miniatuur hout vrugtekratte onder gekontroleerde toestande in die laboratorium te disinfekteer. Na die onderdompeling van die mini vrugtekratte in 'n nematode suspensie van 25 ILs/ml van al drie EPN spesies, onder optimale toestande van temperatuur en humiditeit, is die hoogste persentasie van beheer met die gebruik van S. feltiae (74,85% ± 3.64%) verkry. Die byvoeging van toevoegings middels het S. feltiae se vermoë om te infekteer betekenisvol (p <0,001) tot > 95% verhoog, maar dit het nie tot 'n betekenisvolle toename in die infektiwiteit van H. bacteriophora of S. yirgalemense gelei nie. Die resultate dui daarop dat H. bacteriophora nie 'n geskikte kandidaat is om te gebruik vir die beheer van kodlingmot larwes in besmette hout kratte nie. Die voorkeurkandidaat tans vir beheer is S. feltiae, waarvan die doeltreffendheid verhoog kan word deur middel van die byvoeging van 'n bymiddel. Gedurende die winter wanneer die hele kodlingmot populasie as larwes in diapause is, word geen beheer in boorde toegepas nie. Hierdie studie het getoon dat EPNs in boorde gespuit kan word om sodoende die opkomende kodlingmot populasie na die winter te verlaag en kan ook ingesluit word in 'n GPB program. Die EPNs kan as 'n tweede verdedigingslinie optree en doeltreffende beheer van kodlingmot larwes verseker wat chemiese bespuitings oorleef het, en sodoende beskerming teen weerstandige kodlingmot populasies in die volgende seisoen bied.