关键词: 一串红, SRAP, 体系优化, 品种鉴定

Abstract: The concentrations of primers, dNTP, DNA template, Taq DNA polymerase and annealing temperature were optimized in order to establish SRAP (sequence-related amplified polymorphism) marker system in Salvia splendens and 17 cultivars were identified using the SARP markers established. The suitable SRAP-PCR system was established for Salvia splendens. Totally 25 μL reaction mixture contained PCR buffer (containing 2.0 mmol·L-1 MgCl2), DNA template 60 ng,dNTP mixture 0.2 mmol·L-1,each primer 0.4 μmol·L-1 and Taq DNA polymerase 1 U. Gradient temperature test showed that SRAP-PCR was not sensitive to the variation of annealing temperature (44-56℃) at second stage. Based on the reaction system established, 17 varieties of Salvia splendens were analyzed using 44 SRAP primer combinations and 37 polymorphic primer combinations were screened in total. The PIC (polymorphism information content) value of these markers varied from 0.215 to 0.941, averaging 0.672. The genotypes revealed by each marker ranged from 2 to 17 with average of 6.76. The 17 varieties could be distinguished completely only using a single primer combination F1/R1 and even two varieties quite similar in morphology, Shenzhouhong and Diwang, could be discriminated by several primer combinations, suggesting that SRAP is powerful for variety identification.

Key words: Salvia splendens, SRAP, system optimization, cultivar identification