Differentiating apple sports by pollen ultrastructure : a thesis presented in partial fulfilment of the requirements for the degree of Master of Horticultural Science at Massey University

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Massey University
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Cultivars are plants that form distinct, uniform and stable phenotypes. New cultivars can be protected by Plant Variety Rights (PVR) which allow the owner exclusive rights to the propagation and sale of the plant material. Current PVR identification methods for apple cultivars require detailed records of tree, flower and fruit characteristics to differentiate the new cultivars from known cultivars. This method is slow, expensive and unable to cope with the increasing numbers of sports. Biochemical identification methods such as isozymes, restriction fragment length polymerisation (RFLP), random amplified polymorphism DNAs (RAPD), and minisatellite probes, can quickly and objectively differentiate cultivars, but cannot differentiate apple sports. Previous research suggested that pollen ultrastructure could be an alternative method for plant identification. This thesis is concerned with the development of a technique to differentiate apple sports using pollen exine patterns. Scanning electron microscopy was used to capture images of the apple pollen grain and the exine surface. A digital image analysis algorithm was developed to extract quantitative data from the pollen grain dimensions and pore characteristics, and a Fast Fourier transform extracted quantitative data from the ridge patterns. Statistical methods were applied to the data to differentiate the sports. Pollen harvested from apple flowers in the spring were wider than pollen harvested from flowers forced out of season under artificial conditions. Significant differences between trees were found for pollen grain length:width ratio, percent pore coverage, pore area and pore length but further research is required. However, apple cultivars types 'Red Delicious' and 'Gala' were successfully differentiated by pore and pollen grain variables, and 'Aversang' and 'Ultrared' sports of 'Red Delicious', and 'Splenola' and 'Galalea' sports Of 'Gala' were successfully differentiated by exine ridge patterns and pollen grain measurements. Differentiation of apple sports by pollen requires further development but may be one of the only quick, objective identification methods that can differentiate sports. Sport differentiation would greatly aid PVR establishment and enforcement.
Apples, Breeding, Varieties, Pollen -- Identification