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Construction of a microsatellite based genetic linkage map of almond.

Almond (Prunus dulcis) is the most important nut crop in terms of world production. Due to its health benefit and high nutritional value the consumption and world supply of almond is increasing. To remain competitive in the world market, the Australian almond breeding program was established to produce cultivars with better adaptation to Australian conditions. As part of this program an almond mapping population consisting of 93 F₁ progeny derived from a cross between the American cultivar ‘Nonpareil’ (NP) and the European self-compatible cultivar ‘Lauranne’ (LA) was produced to construct the genetic linkage maps. The first almond linkage map developed prior to the commencement of this project failed to produce the eight linkage groups similar to the basic chromosome number of almond (x = 8) and many large gaps were also observed on the linkage groups. Therefore, more markers were needed to saturate the maps. Microsatellite markers are considered one of the best choices for mapping studies. 195 microsatellite markers isolated from Prunus species were obtained from published papers or by personal communication. Polymorphism was revealed by three different methods, and in general, polyacrylamide gel electrophoresis (PAGE) compared to the fluorescent labelled marker detection using an automated DNA sequencer or agarose gel electrophoresis, showed the most efficient and cost effective method of genotyping. A subset of 54 markers which produced reliable and easily interpretable polymorphic bands was selected to screen the whole mapping population. Microsatellites originally isolated from almond species showed the highest rate of amplification and polymorphism followed by peach microsatellites and the least informative markers were isolated from cherry. It seems that the level of transportability and usefulness of microsatellite markers is related to the genetic distance of the closely related species. Almond and peach belong to the same subgenus (Amygdalus) and other Prunus species are classified in Prunophora subgenus. The nut, or kernel, is the commercial part of the almond tree, thus to improve the quality of fruit an understanding of environmental influence, heritability and correlation of traits is required. Pomological and quality characters such as: shell hardness, kernel size, shape, taste, pubescence, colour, and percentage of doubles were measured during three consecutive years (2005-2007) on the total mapping population, but data analysis (ANOVA) was performed only on trees that survived for all three years. Most of the traits showed high broad-sense heritability and kernel shape showed the highest heritability of H² = 0.92 suggesting high genetic control of this trait. Occasionally larger kernels than either parent were found in the progeny indicating potential for improvement of this trait even with smaller kernel size parent that encompass many desirable characters. High correlation was also found between the in-shell and kernel weight (r = 0.74), kernel length / kernel width (r = 0.67), kernel weight to kernel length (r = 0.78) and kernel width (r = 0.80). This correlation estimation pointed out in this study indicates that the improvement of one character may result the progress in another trait. Neither of the parents in the mapping population had bitter or obvious slightly bitter taste but slightly bitter kernels were observed among the progeny. Amygdalin was assumed to be responsible for bitter taste in almond; therefore we measured the amount of amygdalin in sweet and slightly bitter kernel progeny by HPLC. However, the results showed that amygdalin exists in sweet kernels as well. Although the average amount of amygdalin in slightly bitter kernels (20.34 mg kg⁻¹ FW) was higher than sweet kernels (3.67 mg kg⁻¹ FW), some sweet kernels had higher amounts of amygdalin suggesting the impact of other components on slightly bitter kernel. The highest variability within the traits was observed in the percentage of double kernel, which showed the highest standard error. Strong environmental effects, particularly low temperature at pre-blossom time is speculated to produce much higher double kernels. Three genetic linkage maps, one for each parent and an integrated map were constructed by the addition of 54 new microsatellite markers to the previous dataset. All the data was scored and coded according to the coding system necessary by JoinMap3 which was used for map construction. 131 markers including microsatellite, ISSR, RAPD, SCAR and S-allele markers were placed on the integrated map covering 590.7 cM with the average density of 4.5 cM/marker. The minimum number of six microsatellite markers was placed on linkage group 8 and the linkage group 1 which is the longest linkage group has 14 microsatellite markers. Comparative mapping study with other Prunus maps, especially with the highly saturated reference map showed complete synteny and minor changes in the order of four markers on linkage groups compared with Prunus reference map. The conservation of molecular marker order observed in this study supports the idea of looking at Prunus genome as a single genetic system and practical application of this similarity would be in cross-transportability of microsatellite markers from well developed linkage maps to the less studied species in Prunus. Ten microsatellite loci placed on our map have not been reported before and could be used to improve the density of other Prunus maps, especially the reference map. This study contributed to the better understanding of the mode of inheritance and environmental effect on morphological traits and the effect of amygdalin on kernel taste. The most saturated microsatellite based almond linkage map developed in this study can serve as a framework for future almond breeding program in Australia and benefit Prunus improvement programs internationally. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1348850 / Thesis (Ph.D.) - University of Adelaide, School of Agriculture, Food and Wine, 2008

Identiferoai:union.ndltd.org:ADTP/264598
Date January 2008
CreatorsTavassolian, Iraj
Source SetsAustraliasian Digital Theses Program
Detected LanguageEnglish

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