Identification Of Genetic Relationship Of Almond By RAPD And ISSR

Almond is a kind of economic Macadamia trees. After a long-term evolution and breeding, there were rich varieties. The study on the genetic diversity of almond benefits the collection, conservation, identification and reasonable use of the the germ plasm. In this study, the RAPD and ISSR makers were used to detect the genetic diversity and cluser analysis on wild almonds and 56 cultivars. The cluster analysis and the genetic diversity analysis were applied to study the genetic relationship and diversity of the accessions. The main results were as follows:1. Considering traits of dry sample by slicon gel and all the other factors, the method about extraction and purification of almond genetic DNA was set. The method was simple, high efficient.2. Using the research results of other nut plants, the RAPD and ISSR amplification programs were optimized to set up the reaction system of RAPD and ISSR fitting for analyzing the resources of almond with the orthogonal diagram.3. Ten RAPD primers and fourteen ISSR primers with polymorphic and informative patterns, were selected from a total of 100 RAPD ones and 86 ISSR ones to determine these individuals' genetic diversity. The 23 RAPD primers and 28 ISSR primers generated 91 RAPD fragments and 169 ISSR fragments, respectively. For RAPD marker 84 polymorphic bands were produced, percentage of polymorphic bands was 92.3%; For ISSR marker, 167 polymorphic bands were produced,percentage of polymorphic bands was 98.8%. Jaccard's genetic similarity matrices and of polymorphic bands was 77.1%. All these 61 lines could be clustered into 5 groups by UPGMA method: the first group is Cultivars of china; the second group is cultivars of USA; the third group is wild almond from the north of Xinjiang region in china; the fourth group is three kind of wild almonds from other place of china; the fifth Group is cultivars from Europe.4. There is a good characteristic band for Bianshitou specie. After the amplification the characteristic band was reclaimed, cloned, and got the sequence. Based on the sequence of the characteristic band, SCAR primer pairs were designed and tested. The result showed that the SCAR primer pairs can marker Bianshitou specie.5. Ten cultivars used in our breeding program were analyzed by RAPD, and an RAPD fingerprint map was obtained using 2 polymorphic primers. These lines could be distinguished by combining 2 primers only.