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Citrus is among the most important fruit crops worlstwide, and therefore the preservation and improvement of citrus germplasm is of the essence. Citrus breeders are often faced with the difficulty of distinguishing between new and existing cultivars because of the ambiguous nature of morphological traits due to environmental influences and error in human judgement. The protection of new varieties is very important to the breeder. New varieties cannot be patented in South Africa, but it can be protected by Plant Breeders' Rights, only if it is genetically distinguishable and significantly different economically from existing varieties. Cultivars in four genera (c. sinensis, C. paradisi, C. grandis and C. reticulata) included in the Citrus Improvement Programme (CIP) or cultivars awaiting recognition of Plant Breeders' Rights by the International Union for the Protection of New Plant Varieties (UPOV) were analyzed with Isoenzymes, Restriction Fragment Length Polymorphism (RFLP) and Random Amplified Polymorphic DNA (RAPD). Five enzyme systems (PGM, PGI, MDH, GOT and IDH) were analyzed and founded to be suitable for grouping together cultivars belonging to the same genera. It was not suited for routine discrimination of cultivars in a particular genus. RFLP studies were conducted on five grapefruit cultivars, using cDNA clones from a genomic library of Rough Lemon. RFLP studies were valuable for the discrimination of closely related cultivars which probably originated from a common ancestor by bud mutations. This technique was, however, abandoned due to its biohazardous nature and replaced by the PeR-based Random Amplified Polymorphic DNA. RAPDs are easy to perform and gave promisin& results which were exploited to reveal polymorphisms between cultivars within the various groups. Although the interpretation of data produced by this method is often suspicious, it is the best method currently available for cultivar identification. It can playa complementary role in the protection of new varieties when classical morphological interpretation of differences is not capable of determining sufficient distinctness for the awarding of Plant Breeders' Rights.

Thesis (M.Sc. (Plant protection)) --University of Limpopo, 2013. / Studies were initiated to investigate (1) the aggressiveness of the citrus nematode (Tylenchulus semipenetrans Cobb) isolates from two provinces in South Africa (2 experiements ) and (2) the biotype of T. semipenetrans in South Africa. In the aggressive study, isolates from Limpopo and Mpumalanga Provinces were used on Carrizo citrange (Citrus sinensis × Poncirus trifoliata) and rough lemon (Citrus jambhiri) seedling rootstocks under greenhouse conditions (18 experiements). Each seedling was inoculated with 0, 10 000, 20 000, 30 000 and 40 000 J2s of T. semipenetrans isolates, which were arranged in a randomised complete block design, with six replications. At 120 days, the reproductive factor of T. semipenetrans isolate from Mpumalanga Province was significantly higher than that from Limpopo Province. Similarly, due to its higher relative impact on the reproductive factor values, the Mpumalanga isolate reduced plant growth variables more than the Limpopo isolate. Consequently, the Mpumalanga isolate was viewed as being more aggressive than the Limpopo isolate, suggesting that there might be genetic variability and/or adaptation in populations from the two locations. A national study, comprising T. semipenetrans isolates from 18 citrus-producing district municipalities in South Africa was then initiated under greenhouse conditions using isolates from each district – for a total of 18 separate experiments. Three differential hosts, viz. rough lemon, P. trifoliata and olive (Olea europaea), served as treatments, arranged in a randomised complete block design, with 15 replications. Initially, an orchard was randomly selected in each of the six citrus-producing provinces, viz. Eastern Cape, KwaZulu Natal, Limpopo, Mpumalanga, North West and Western Cape. Three-month old differential host seedlings were inoculated with approximately 10 000 J2s of T. semipenetrans and allowed to establish and grow under greenhouse conditions. At 120 days, penetration indices and standardised reproductive potentials/g roots demonstrated that T. semipenetrans failed to reproduce and develop on olive, but reproduced and developed on the other two hosts. Using T. semipenetrans biotype classification system, findings suggested that the biotype in citrus-producing district municipalities was Poncirus biotype. This biotype reproduces on P. trifoliata and hybrid rootstocks, which therefore, suggested that trifoliate orange and its hybrid rootstocks were not suitable for use in managing population nematode densities of T. semipenetrans in South Africa. In conclusion, results of this study demonstrated that the South African T. semipenetrans biotype was Poncirus, which suggested different management decisions and strategies for the citrus industry with regard to the management of this nematode. / By National Research Foundation, National Department of Agriculture, Forestry and Fisheries and the Land Bank Chair of Agriculture − University of Limpopo,