中文摘要：

EcoTILLING （Ecotype Targeting Induced Local Lesions IN Genomes）可快速检测自然群体中的SNP（Single Nucleotide Polymorphism）等DNA变异。第一次建立了简化的茶树EcoTILLING技术体系：首先筛选扩增目的基因的特异PCR引物；然后通过CELI酶切PCR产物优化实验，确定酶切时间20min、10×buffer组成为250mmol／L Tris-HCI（pH7．4）、150mmol／LKCI、15mmol／LMgSO4、4lag／mLBSA和0．04％Triton x-100，反应温度为47．5℃，酶量为1．8μL CJE（Celery Juice Extract）／10μL反应体系时，酶切的谱带信号较强、特异性高；初步确定合适的非变性聚丙烯酰胺凝胶电泳检测条件为凝胶浓度5％、上样量4～5μL、有效分离长度要达到15cm。利用上述简化的EcoTILLING对6份茶树种质870bp的PPO基因片段实现了分型，经测序验证酶切产生的谱带与6份种质内部或不同种质间的SNP位置吻合。

英文摘要：

EcoTILLING（Ecotype Targeting Induced Local Lesions In Genomes） had been used as a SNP（Single Nucleotide Polymorphism） discovery tool to examine DNA variation in natural populations. A simple EcoTILLING technique for tea plant was established. Firstly, specific PCR primers for the amplification of the target gene were screened out. Then through comparison, a fine CELI enzyme digestion could be resulted using 20min with 10× buffer containing 250 mmol/L Tris-HC1 （pH7.4）, 150 mmol/L KCI, 15 mmol/L MgSOa, 4 μg/mL BSA and 0.04%Triton x-100, and 1.8 μL CJE（Celery Juice Extract）/101aL reacting solution at 47.5℃. Moreover, a good picture could be obtained when using 5% polyacrylamide gels （PAGE）, adding 4~5 laL digesting products in PAGE electrophoresis. The 870 bp fragment within the PPO （Polyphenol Oxidase） gene of 6 tea germplasms was distinguished from each other using the simplified EcoTILLING. The CEL I digested bands were ascertained that according to the position of SNP in or between the 6 accessions through sequencing verification.