Establishment of ISSR Reaction System for Armeniaca sibirica Based on Orthogonal Design  

作　　者：Jinjun BAI 

机构地区：[1]College of Chemistry and Chemical Engineering, Yulin University

出　　处：《Agricultural Biotechnology》2012年第3期1-3,6,共4页农业生物技术（英文版）

基　　金：Supported by Start-up Fund for High-level Scientific Research Personnel(11GK30)

摘　　要：[ Objective] The aim was to establish the optimal ISSR-PCR system for Armeniaca sibirica. [ Method] With Armeniaca sibirica as experimental materi- al, genomic DNA was extracted from tender leaves using CTAB method. Orthogonal experiment L9 （34） was conducted with four factors （Mg2＋ , dNTPs, primers, Taq DNA polymerase） in ISSR-PCR system ofArmeniaca sibirica at three levels. The results of orthogonal experimental design were analyzed by using SPSS statisti- cal software. [Result] Different levels of various factors all had significant effects on PCR results, specifically, the amount of Taq DNA polymerase had the most significant effect. The optimal ISSR-PCR reaction system of Armeniaca sibirica was established ： the total PCR reaction volume was 20 ~1, containing 1.0 U of Taq DNA polymerase, 1.0 mmol/L MgCI2,0.15 mmol/L dNTPs and 0.30 ~mol/L primers. Validation result shows that this reaction system is very stable and strongly operational. [ Conclusion] This study laid the foundation for genetic diversity analysis and germplasm resource investigation of Arnniaca sibirica.[ Objective] The aim was to establish the optimal ISSR-PCR system for Armeniaca sibirica. [ Method] With Armeniaca sibirica as experimental materi- al, genomic DNA was extracted from tender leaves using CTAB method. Orthogonal experiment L9 （34） was conducted with four factors （Mg2＋ , dNTPs, primers, Taq DNA polymerase） in ISSR-PCR system ofArmeniaca sibirica at three levels. The results of orthogonal experimental design were analyzed by using SPSS statisti- cal software. [Result] Different levels of various factors all had significant effects on PCR results, specifically, the amount of Taq DNA polymerase had the most significant effect. The optimal ISSR-PCR reaction system of Armeniaca sibirica was established ： the total PCR reaction volume was 20 ~1, containing 1.0 U of Taq DNA polymerase, 1.0 mmol/L MgCI2,0.15 mmol/L dNTPs and 0.30 ~mol/L primers. Validation result shows that this reaction system is very stable and strongly operational. [ Conclusion] This study laid the foundation for genetic diversity analysis and germplasm resource investigation of Arnniaca sibirica.

关 键 词：Armeniaca sibirica PCR ISSR Orthogonal design 

分 类 号：S662.2[农业科学—果树学]