Study On Germplasm Resources Of Pear In The Fujian Province By ISSR Technique

Pear belongs to Rosaceae, Maloideae, Pyrus L. The Chinese pear have been cultivated more than 3,000 years of history, the pear species in the world there are about 30 species, There are 13 kinds of which originated in China. In order to understand the origin of our species of pear genus Pyrus in the whole of the location and the origin of hybrid pear. It's necessary to use the latest methods of molecular biology, the relationship between plant species and the origin of China's pear-depth study done.ISSR molecular marker techniques developed a new molecular biology technology over the past ten years, has been widely used in mammals and higher crop-related research in the field. With this study, ISSR molecular markers for 56 pear cultivars the level of a DNA study aimed at establishing a simple, rapid, high-quality pear extract and purify DNA technology,And the establishment of an appropriate technical system ISSR pear, as well as research on the species cluster analysis, genetic relationship, genetic diversity analysis, Have provided the deeper more important theory basis for the next pear heredity multiplicity and the variety appraisal. The main result is as follows:1. Ripe pear leaves for tannin-rich, polysaccharide, pigments and phenolic secondary metabolites, Uses the hot boric acid law to withdraw DNA, the content is high, does not have disturbance materials and so on protein, RNA or polyphenols, Regardless of the quantity or purity can meet the requirements of ISSR analysis, has established the suitable pear DNA extraction method.2. Through system's research quite, has established the stable pear ISSR-PCR analysis reacting system. The reacting system is: in 20μl systems including pattern plate DNA30 ng, directs the density is 0.4μM, the dNTPs density is 0.20 mM, TaqDNA polymerase amount used is 1 U, 10×Buffer 2 L. Amplification procedure is: 94℃predegeneration 5 min, 94℃denaturation l min, 52℃annealing 1 min, 72℃ extension of 2 min, 40 cycles; 72℃extension 8min, 4℃stored. This system increases the banding is clear, the effect is good, can satisfy the ISSR analysis.3. Primers from 96 of 11 selected ISSR primers, of 56 Pear materials ISSR analysis, A total of 127 ISSR amplified bands, the average of each amplified band of 11.5. A total of 11 primers amplified 113 polymorphic loci, the average of each amplified 10 loci, percentage of polymorphic loci 87.3 %.4. The basis of ISSR analysis for the examination of 56 pear varieties cluster analysis,The results show that,the genetic similarity between 0.513-0.868, average 0.667, Revealed the 56 pear cultivars in Fujian Province of the genetic relationship between, for the future genetic diversity and pear variety identification is more important to provide a deeper theoretical basis.