Identification of molecular markers linked to woolly apple aphid (Eriosoma lanigerum) (Hausmann) resistance in apple

Christians, Gillian Eleanore (2003-12)

Thesis (MSc)--Stellenbosch University, 2003.

Thesis

ENGLISH ABSTRACT: Apple (Malus x domestica Borkh.) is an important horticultural crop worldwide and in the Western Cape. The income generated from apple and other deciduous fruit production amounts to approximately 25% of the gross total value of horticultural production in the Western Cape. Unfortunately diseases and pests adversely affect fruit production in this region. Woolly apple aphids (Eriosoma lanigerum L. (Hausmann» have a significant effect on the apple industry in the Western Cape. Damage caused is two-fold, occurring aerially and terrestrially. Insects colonise the plants, feeding off the phloem sap. Aphid infestation around the root system results in repeated infestation of the foliage as it serves as a reservoir of aphids. In extreme cases, the apple cores are also infested, thus affecting the sale of apples. In 1962, Northern Spy was identified as a woolly apple aphid resistant rootstock and has since then formed the basis for traditional rootstock breeding programmes. The Er1 gene in Northern Spy confers resistance. According to one report, the natural resistance of Northern Spy was overcome in South Africa in 1968, but this was not confirmed in an independent study. The main objectives of this study was to firstly identify molecular markers more closely linked to the woolly apple aphid resistance gene, Er1, than existing markers, by applying AFLP technology to selected seedlings, identified to be resistant by conventional phenotyping. If identified, these markers can be incorporated into existing breeding programmes. Secondly, previously identified RAPD and SCAR markers were tested to determine their applicability in local populations for use in breeding programmes. Ultimately the segregation of the Er1 gene in South African populations can be determined if tightly linked markers are identified. Three families were derived from crosses of each of three resistant genotypes, namely Northern Spy, Rootstock 5 and Russian Seedling and a susceptible cultivar, Braeburn. For the three successive years of the study, each resistant genotype was allowed to cross-pollinate in isolation with the susceptible parent. Two hand-pollinated families, Russian Seedling x Liberty and Russian Seedling x Northern Spy, were also included in the study. The amplified fragment length polymorphism (AFLP) technique was used in an attempt to identify markers in the resistant and susceptible seedlings. No markers were identified using this technique. Known sequence characterised amplified regions (SCAR) and random amplified polymorphic DNA (RAPD) markers were used due to their suitability in marker-assisted selection for woolly apple aphid resistance. Varying results were obtained with these markers and no conclusive information was acquired with regard to the segregation of the Er] gene in any of these rootstocks and crosses. This underlines the need for the development of markers that can readily be applied in local breeding programmes. The identification and integration of such markers will greatly benefit the local and world wide apple industries.

AFRIKAANSE OPSOMMING: Appels (Malus x domestica Borkh.) is wêreldwyd en in die Wes-Kaap 'n belangrike landbougewas. Inkomste gegenereer deur appels en ander sagtevrugte vorm bykans 25% van die bruto inkomste uit vrugte in die Wes-Kaap. Siektes en insekpeste verlaag egter die produksie van vrugte in hierdie streek. Appelbloedluise (Eriosoma lanigerum L. (Hausmann» het 'n groot invloed op appelproduksie in die Wes-Kaap. Skade word bogronds en ondergronds aangerig. Insekte koloniseer die plant en leef op floeëmsap. Besmetting van die wortels lei tot herhaalde besmetting van bogrondse dele aangesien die insekte aanteelop die wortels. In uiterste gevalle word die vrugte geaffekteer, wat vrug-verkope beïnvloed. 'Northern Spy' is in 1962 geïdentifiseer as 'n onderstam met natuurlike weerstand teen appelbloedluis en het vir lank die basis gevorm vir tradisionele telingsprogramme. Weerstand word verleen deur die Erf geen. Volgens een verslag is die natuurlike weerstand van Northern Spy egter in 1968 in Suid-Afrika oorkom, maar dit is nog nie in 'n onafhanklike studie bevestig word nie. Die hoof doelstellings van hierdie studie was om eerstens deur middel van die AFLP tegniek molekulêre merkers te identifiseer wat nouer gekoppel is aan die appelbloedluis weerstandsgeen, En, as bestaande merkers. Hierdie tegniek is toegepas op saailinge wat deur konvensionele fenotipering geselekteer is. Indien merkers suksesvol geïdentifiseer is, kan dit in bestaande telingsprogramme geïntegreer word. Tweedens is bestaande RAPD en SCAR merkers ook getoets om hul toepaslikheid te bepaal vir gebruik in plaaslike teelprogramme. Oplaas sal die segregasie van die Erf geen in Suid- Afrikaanse populasies ook deur middel van nou gekoppelde merkers bepaal kan word. Kruisings van elk van die drie weerstandbiedende genotipes, naamlik 'Northern Spy', 'Rootstock 5' en 'Russian Seedling', en die vatbare kultivar, 'Braeburn' , het drie families daargestel. Elke weerstandbiedende genotipe is toegelaat om in isolasie te kruisbestuif met die vatbare ouer. Twee hand-bestuifde families, 'Russian Seedling' x 'Liberty' en 'Russian Seedling' x 'Northern Spy', is in 'n latere stadium van die studie ingesluit. Die AFLP tegniek is gebruik vir die identifikasie van polimorfiese merkers tussen vatbare en weerstandbiedende populasies. Geen merkers is egter geïdentifiseer nie. Bestaande SCAR en RAPD merkers is vervolgens gebruik om te bepaal of hulle geskik is vir gebruik in merker-bemiddelde seleksie vir appelbloedluis weerstand. Wisselende resultate is verkry ten opsigte van amplifikasie, herhaalbaarheid van resultate was swak en geen onweerlegbare bewyse oor die segregasie van die Erfgeen is bekom nie. Dit beklemtoon die noodsaaklikheid om merkers wat geredelik in plaaslike teelprogramme toegepas kan word, te ontwikkel. Die identifikasie en integrasie van sulke merkers sal die plaaslike en wêreld-wye appel industrieë aansienlik bevoordeel.

Please refer to this item in SUNScholar by using the following persistent URL: http://hdl.handle.net/10019.1/53454
This item appears in the following collections: