庞宏光;许建锋;张江红;张玉星;亓宝秀;

• 1:河北农业大学园艺学院
• 2:北京林果业生态环境功能提升协同创新中心

摘要(Abstract)：

【目的】针对杜梨组织中富含多酚、多糖等次生代谢物质特点,筛选从杜梨不同组织中提取高质量RNA的方法。【方法】以杜梨不同组织(根、茎、叶、花和果)为材料,分别采用Trizol法、改良CTAB法和RNAprep Pure Plant Kit试剂盒法提取总RNA,利用琼脂糖凝胶电泳和NanoDrop 2000微量分光光度计对RNA进行质量检测。【结果】3种方法均能提出杜梨不同组织中总RNA,但RNAprep Pure Plant Kit法提取的总RNA电泳条带清晰、完整性较好,根、茎、叶、花和果提取的RNA纯度分别为2.07、2.02、2.01、2.01和2.02;浓度分别为186.2 ng·μL~(-1)、174.5 ng·μL~(-1)、163.6 ng·μL~(-1)、167.3 ng·μL~(-1)和121.1 ng·μL~(-1)。以RNAprep Pure Plant Kit试剂盒法提取的总RNA反转录成cDNA为模板,内参基因PbUBI为目的基因进行RT-PCR扩增,均可获得明亮单一条带。【结论】RNAprep Pure Plant Kit试剂盒法更适用于杜梨不同组织总RNA提取,可为下一步进行基因克隆及功能验证等分子生物学研究奠定基础。

关键词(KeyWords)： 杜梨;不同组织;总RNA;纯度;浓度

Abstract:

Keywords:

基金项目(Foundation): 科技创新服务能力建设-科研基地-林果业生态环境功能提升协同创新中心(CEFF-PXM2018_014207_000024)

作者(Author): 庞宏光;许建锋;张江红;张玉星;亓宝秀;

Email:

DOI: 10.13925/j.cnki.gsxb.2018.S.11

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