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Generation of genotyping profile in comm Generation of genotyping profile in comm... - Complex Object ()
Generation of genotyping profile in commercial stone fruits to aid forensic investigations / Aishah Kadher
[Truncatedabstract] The Prunus genus is important because of its high nutritional andculinaryattributes. The fruits belonging to this genus are commonly known asstone fruits or drupes. Thecultivation of stone fruits hasdrivenregional economies. Even though thestone fruits rank third as an economically significant produce, the trade remains principally unconstrained. Deceitful practices by vendors toboost their turnover such as substituting and adulterating high quality fruits with cheaper varieties goes unchecked. Consumers’ rightsare violated as they are cheated by unscrupulous vendors who sell badquality fruits for the price of goodones. Furthermore,the orchardist who developedthe fruit is not rewarded for efforts as royalties are not paid.Hence to avoid commercial exploitation and to protect plant breeders’ rights, it has become vital to authenticate fruit varieties in orderto minimise the incidence of illegalactivity. The traditional techniques used forthe identification of the genus Prunus are based on morphological, physiological andchemical characteristics of the fruits. However, these techniqueshave limitations. For example,multiple names for the same fruit variety canbe assignedand ambiguous inputsin breeding programs have resulted in misinterpretation of descriptiveterms used. Further, physical traits are dependent on environmental factors and thus will vary between different orchards. Consequently, other techniques for complete authentication of the fruits and other plant tissues are required. DNA profiling provides a technique suitable for analysing various fruit varieties, since the DNA within different fruits of the same variety does not change. Commercially sold stone fruit varieties are either of low quality or processed in a way that may lead to degraded DNA in an analytical sample, thus making extraction and amplification difficult.
In this project, the CTAB (Cetyl tri methyl ammonium bromide) method, together with three other nucleic acid extraction techniques, were used to extract DNA from different parts of fruits and seeds of the Prunus genus. The purpose of using different techniques was to determine the reproducibility and quality of the results obtained. The DNA extracted by the CTAB method was successfully amplified using primers selected for this project. The primers used in this study generated adequate bands to differentiate among closely related varieties. The outcome of this study indicates both that the microsatellite technique provides a comparatively rigorous way of examination and also that assays can be fully programmed for high throughput analysis. With the help of this technique it is possible to distinguish closely associated Prunus varieties from one another. Six microsatellites (Pchgms 12, Pchgms 20 F1, Pchgms 20 F2, BPPCT 026, UDP 96-003 and UDP 96-015) were used in this research. The Pchgms series of microsatellites enabled to study the level of polymorphism in stone fruit varieties. The Genescan results of the exocarp, mesocarp and endocarp of all fruit varieties, generated after amplification were compared. The aim of this comparison was to show that the DNA profiles of the mesocarp and exocarp were similar to each other, as these tissues in principle inherit alleles from the maternal parent (flower)...
Thesis (M.For.Sc.)--University of Western Australia, 2010
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