【摘要】 目的:优化并验证辣椒“CMS”三系多重PCR分子标记。方法:以改良CTAB法提取的DNA为模板，优化与辣椒“CMS”三系相关的多重PCR分子标记体系，并对20份辣椒种质材料进行育性鉴定，以验证其准确性。结果:优化后20μL的PCR体系含10μL Mix、1μL引物SCAR_130/140、1μL CRF-SCAR、1μL DNA模板，ddH₂O补齐；优化的PCR扩增程序为94℃预变性3 min, 94℃变性30 s, 57.8℃复性30 s, 72℃延伸90 s, 35个循环，最后72℃延伸5 min, 4℃保存。利用优化后的多重PCR对A、B、C三系材料进行验证，准确率分别是85.7%、100%和80%。结论:优化后的多重PCR反应体系重复性好、稳定可靠。更多还原

【Abstract】 Objective: To optimize and validate the molecular marker of "CMS" triplex PCR in pepper. Methods: The DNA extracted by the modified CTAB method was used as a template to optimize the multiplex PCR molecular marker system related to the three lines of "CMS" in pepper, and the fertility identification of 20 pepper germplasm materials was carried out to verify the accuracy of the optimized multiplex PCR. Results: The optimized 20 μL PCR system included 10 μL of PCR Mix, 1 μL of primer SCAR_130/140 and 1 μL of CRF-SCAR, 1 μL of DNA template, and the insufficient part was supplied by ddH₂O. The optimized PCR amplification procedure was pre-denatured at 94 ℃ for 3 min, denaturation at 94 ℃ for 30 s, renaturation at 57.8 ℃ for 30 s, extendation at 72 ℃ for 90 s with 35 cycles, then extended at 72 ℃ for 5 min at last and stored at 4 ℃. The optimized multiplex PCR was used to verify the A, B, and C materials, and the accuracy rates were 85.7%, 100% and 80%, respectively. Conclusion: The optimized multiplex PCR reaction system has good repeatability, stability and reliability.更多还原