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对影响枣ISSR反应的相关因子进行了研究。优化的反应体系包含:2.5μL 10×buffer,50 ng模板DNA,2.5 mmol.L-1 Mg2+,0.3 mmol.L-1 dNTPs,0.3μmol.L-1引物,2 U Taq DNA聚合酶,反应体积为25μL;优化的PCR扩增程序为:94℃2 min;94℃l min,退火1 min,72℃2 min,循环35次;72℃延伸10 min;退火温度根据引物温度设定。该ISSR体系被用于17个枣栽培品种遗传图谱分析,结果表明所构建的遗传图谱能较好地反应供试品种的遗传多样性。
Relevant factors affecting the jujube ISSR response were studied. The optimized reaction system consisted of 2.5 μL 10 × buffer, 50 ng template DNA, 2.5 mmol.L-1 Mg2 +, 0.3 mmol.L-1 dNTPs, 0.3 μmol.L-1 primer, 2 U Taq DNA polymerase, reaction volume The optimal PCR procedure was as follows: 94 ℃ for 2 min; 94 ℃ for 1 min, annealing for 1 min, 72 ℃ for 2 min, cycling for 35 cycles; extension at 72 ℃ for 10 min; annealing temperature was set according to the primer temperature. The ISSR system was used for the genetic analysis of 17 jujube cultivars. The results showed that the constructed genetic map can better reflect the genetic diversity of the tested cultivars.