|Table of Contents|

Establishment of ISSR Reaction System in Quercus variabilis (PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2014年13期
Page:
93-97
Research Field:
Publishing date:

Info

Title:
Establishment of ISSR Reaction System in Quercus variabilis
Author(s):
LI Na; ZHANG Cun-xu; CUI Xiao-yan; JIANG Hong-gang; GUO Le; LIU Wei
College of Forestry，Northwest Agricultural and Forestry University，Yangling,Shaanxi 712100
Keywords:
Quercus variabilis; ISSR; system optimization; genetic diversity
PACS:
S 792.189
DOI:
-
Abstract:
ISSR reaction conditions of Quercus variabilis were optimized systematically via single-factor experiment by using the tender leaves of Quercus variabilis as test materials，resulting in establishment of optimal ISSR reaction system and thermal cycling conditions of Quercus variabilis.The results showed that the optimal ISSR reaction system included a total volume 25 μL containing 2.5 μL 10×Buffer，30 ng template DNA，1.5 U Taq DNA polymerase，2.5 mmol/L MgCl2，0.6 μmol/L primer，0.3 mmol/L dNTPs.Thermal cycling conditions were as follows：denaturation 1 min at 94℃；35 cycles of 30 s at 94℃，60 s at 51.5～59.0℃，2 min at 72℃；and a final extension of 5 min at 72℃ at the end of the amplification.Based on the optimized reaction system，12 primers with superior stability and polymorphism were selected from 100 ISSR primers.166 bands were amplified in 22 individuals of the natural population in Shaanxi Province,among which 142 were polymorphic loci,the percentage of which was 85.54％,Nei’s index 0.2928 and Shannon’s information index was 0.4381,indicating the relatively higher genetic diversity within population of Quercus variabilis.

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Last Update: 2014-08-02