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Establishment of SSR Reaction System and Analysis of Genetic Diversity of Apple Cultivars

Author: XuXingXing
Tutor: YangMinSheng
School: Agricultural University of Hebei
Course: Tree Genetics and Breeding
Keywords: Apple SSR markers Species identification Genetic Diversity
CLC: S661.1
Type: Master's thesis
Year: 2006
Downloads: 351
Quote: 4
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Abstract

Apple is the world's most widely cultivated fruit trees, because Apple has a child of a long, long growth cycle, plants Japanese self-incompatibility and other characteristics, using traditional methods of apple cultivar identification time consuming and expensive. In recent years, with the rise of molecular biology, especially DNA molecular markers in the development and application of apple, apple varieties make it possible to quickly identify which SSR markers for its rich polymorphism, codominant good reproducibility and easy operation increasing people's attention. In this study, to establish and optimize Apple SSR reaction system, based on the application of SSR markers 42 apple varieties on the genetic diversity has been studied at the molecular level for the apple germplasm and varieties of kinship analysis provides a basis for , and SSR markers used in the identification and variety protection apple varieties were evaluated feasibility. The main results are as follows: (1) a reaction of the apple varieties SSR system optimized to determine the optimum reaction system: In 20μL reaction system, Mg ^{2 , primers, and the optimum concentration of dNTP were 2.5mmol · L ^{-1 , 0.8μmol · L ^{-1 , 0.10mmol · L ^{-1 ; TaqDNA polymerase reaction system should join 1U, template DNA should be added to 15 ~ 30ng. PCR reaction thermal cycling program: 94 ℃ denaturation 4min; 94 ℃ denaturation 1min, 48.5 ~ 52 ℃ annealing 1min, 72 ℃ extension 1min, 30 cycles: 72 ℃ for 5min. After repeated amplification tests show that the system repeatability and stability. (2) In this study, 12 pairs of primer combinations 42 apple varieties were amplified by PCR, amplified a total of 165 DNA bands were polymorphic with 90, accounting for the total number of bands amplified for 54.88%, the different primer with distribution of 5 to 24 with an average of 13.6. SSR bands according to the statistical results, application software NTSYS genetic similarity coefficient calculation, 42 apple varieties similarity coefficients ranged from 0.713 to 1.000, which indicated the test apple germplasm genetic variation among little genetic diversity resistance is low. Was constructed using the UPGMA dendrogram, results showed that: the genetic distance of 0.800 when the threshold SSR markers can well distinguish all outside except for Fuji varieties (lines), will be divided into four groups of apple varieties , most of the varieties of the same or similar pedigree gathered together. Visible, SSR molecular marker technology by means of conventional cross-breeding of cultivars to distinguish, but can better reveal the genetic relationship among varieties. (3) the use of polyacrylamide gel (8%) technology on different apple varieties tested primer amplification products were detected, obtained in a group of each species SSR locus genotype patterns, which initially established an apple cultivars tested SSR genotypes atlas for the establishment of a database of apple varieties with SSR genotypes laid the foundation, but also proved SSR markers used in apple cultivars identification and variety protection is feasible.}}}}

Establishment of SSR Reaction System and Analysis of Genetic Diversity of Apple Cultivars

Abstract

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