An orthogonal design was used to optimize ISSR reaction system for Leptosphaeria biglobosa on oil- seed rape in five factors(Taq polymerase，Mg2 +，dNTPs and the concentration of primer or template) at four levels． A suitable ISSR- PCR reaction system for Leptosphaeria biglobosa was established．The optimized reaction system consists of 1 U Taq DNA polymerase，2． 0 mmol /L Mg2 +，0． 2 mmol /L dNTPs，0． 6 mol /L primer，40 ng template DNA，and 2． 5 L 10 × Buffer．24 primers were selected for the abundant and clear band from 110 random primers and their optimal annealing temperature was decided．This optimization of ISSR- PCR reaction system can be used to study the genetic diversity of the pathogen which was confirmed by testing on the stability of 20 strains of Leptospha- eria biglobosa．
HAO Li-fen
,
SONG Pei-ling
,
LI Zi-qin
,
LI Xin-zhou
,
ZHANG Jian
,
Bao Yu-ying
. Optimization for ISSR- PCR Reaction System on Leptosphaeria biglobosa[J]. Acta Agriculturae Boreali-Sinica, 2013
, 28(4)
: 204
-207
.
DOI: 10.3969/j.issn.1000-7091.2013.04.037
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