OPTIMIZATION OF THE INTER-SIMPLE SEQUENCE REPEATS REACTION SYSTEM IN LOQUAT GERMPLASM

The optimization of ISSR (Inter-simple Sequence Repeats) in Ullera (Eriobotrya japonica Lindl. cv. ‘Ullera’) was investigated with primer AGAGAGAGAGAGAGAGYC. The results showed that in 25 µl reaction system of ISSR, the optimum concentrations of five important components Taq DNA polymerase, Mg²⁺, primer, template DNA and dNTPs were 1.2 U, 3.0 mmol L^-1, 0.24 µmol L^-1, 20 ng and 0.15 mmol L^-1, respectively. Using the optimized system, 48 germplasms of Eriobotrya from USA, Italy, Spain, Japan, Algeria and China were studied with the ISSR technique and AGAGAGAGAGAGAGAGYC as primer. We obtained clear ISSR amplification profile by agarose gel electrophoresis.