Karakterisering van derivate uit 'n Thinopyrum distichum X tetraploïede rog kruising

Jacobs, Johan Adolf (2002-03)

Thesis (MSc)--University of Stellenbosch, 2002.

Thesis

ENGLISH ABSTRACT: Soil salinity is a major limiting factor of plant and crop growth, because the absorption of water and nutrients is such a complex process while low and moderate salinity are omnipresent. Plant growth is affected negatively if a specific ion concentration exceeds its threshold and becomes toxic. The detrimental effect of soil affected by salt on crop production is increasing worldwide (Tanji, 1990). The level to which plants can tolerate high salinity levels is genetically controlled with several physiological and genetic mechanisms contributing to salt tolerance (Epstein & Rains, 1987). The most effective way of addressing the limitations of crop productivity in saline areas, is the development of salt tolerant varieties. Understanding the genetics of salt tolerance is, therefore, necessary for the development of an effective breeding strategy for salt tolerance. The department of Genetics (US) conducts a wide crosses research programme aiming to transfer genes for salt tolerance to wheat and triticale. The donor species, Thinopyrum disticum, an indigenous coastal wheat grass, adapted to high concentrations of salt, was crossed with cultivated rye (Secale cereale) in an attempt to study the genetics of salt tolerance (Marais et al., 1998). The primary goal of this study was to find molecular markers (RAPD and AFLP) which associate with chromosomes promoting salt tolerance for later attempts to transfer the genes to triticale. Seventy clones of secondary hybrids (Th disticum /4x-rye 1/2x-rye) were tested for salt tolerance and showed different levels of salt tolerance. RAPD-marker analyses were used to identify polymorphisms between salt tolerant and salt sensitive plants. Twelve RAPD primers produced clear, analyzable and repetitive polymorphic . fragments that can be used as useful markers. Different AFLP-primer combinations were tested against the genotypes of 15 clones (Marais & Marais 2001, unpublished data) and produced approximately 2000 clearly distinguishable AFLP fragments, of which 54 (3%) were polymorphic fragments. Two RAPD fragments and 4 AFLP fragments that can be used as possible markers for the presence of chromosomes that contribute to salt tolerance were identified. The interpretation of the markers was complicated by heterogeneity among plants with regard to the origin of their chromosomes and the genetic diversity of the rye genome. It is also possible that chromosome re-arrangement took place during backcrossing, which could have complicated the data.

AFRIKAANSE OPSOMMING: Versouting is een van die groot beperkende faktore op plant- en gewasgroei, omdat die opname van water en voedingstowwe so In ingewikkelde proses is en die effek van lae of matige versouting so alomteenwoordig is. Plantgroei word nadelig geaffekteer as 'n spesifieke ioonkonsentrasie sy drempelwaarde oorskry en toksies word. Die nadelige effek van soutgeaffekteerde grond op gewasproduksie, is wêreldwyd aan die toeneem (Tanji, 1990). Die vlak waartoe plante hoë konsentrasies sout kan hanteer is onder genetiese beheer met verskeie fisiologiese en genetiese meganismes wat 'n bydrae maak tot soutverdraagsaamheid (Epstein & Rains, 1987). Die mees effektiewe manier om die beperkinge op gewas produktiwiteit in versoute gebiede te oorkom, is die ontwikkeling van soutverdraagsame variëteite. Begrip van die genetika van soutverdraagsaamheid is dus noodsaaklik vir die ontwikkeling van In effektiewe telingsstrategie. Die departement Genetika (US) bedryf tans 'n wye-kruisings navorsingsprogram waarmee gepoog word om gene vir soutverdraagsaamheid na korog en koring oor te dra. Die skenkerspesie, Thinopyrum disticum, In inheemse strandkoringgras wat aangepas is by hoë konsentrasies sout, is gekruis met verboude rog (Secale cereale) in 'n poging om die oorerwing van soutverdraagsaamheid te bestudeer (Marais et al., 1998). Die hoofdoel van hierdie studie was om molekulêre merkers (RAPD en AFLP) te vind, wat assosieer met chromosome wat soutverdraagsaamheid bevorder en om nuttige merkers daar te stel vir latere pogings om die gene na korog en koring oor te dra. Ongeveer 70 klone van sekondêre hibriede (Th distichum I 4x-rog /I 2x-rog) is onderwerp aan souttoetse en het verskillende grade van soutverdraagsaamheid getoon. RAPDmerker analise is gebruik om polimorfismes te identifiseer tussen soutverdraagsame en soutsensitiewe plante. Twaalf RAPD inleiers het duidelike, ontleedbare en herhalende polimorfiese fragmente opgelewer en moontlike nuttige merkers uitgewys. Verskillende AFLP-inleier kombinasies, wat getoets is teen die genotipes van 15 klone (Marais & Marais, 2001 ongepubliseerde data) het ongeveer 2000 duidelik onderskeibare AFLP fragmente geproduseer, waarvan 54 (3%) polimorfiese fragmente was. Twee RAPD fragmente en 4 AFLP fragmente is geïdentifiseer wat as moontlike kandidaat merkers gebruik kan word vir die identifisering van chromosome wat bydra tot soutverdraagsaamheid . Die interpretasie van die merkers is bemoeilik deur heterogeniteit tussen die plante wat betref die agtergrond van chromosome wat hulle besit en die genetiese diversiteit van die rog genoom. Dit is ook moontlik dat chromosoom herrangskikking plaasgevind het tydens terugkruising, wat die data verder kon kompliseer.

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