| 张子轩，郭融融，次而甲玛，王宏鹏，王俊斌，曹高燚，包曙光，谢晓东，陈小强.适用于转录组测序的大麦胚芽鞘保卫细胞分离方法优化[J].麦类作物学报,2021,(4):410 |
| 适用于转录组测序的大麦胚芽鞘保卫细胞分离方法优化 |
| Optimization of Guard Cell Isolation from Barley Coleoptiles for Transcriptome Sequencing |
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| DOI： |
| 中文关键词: 大麦 胚芽鞘 保卫细胞 酶解 |
| 英文关键词:Barley Coleoptile Guard cell Enzymolysis |
| 基金项目:天津市大学生创新创业训练计划项目（201910061055）；国家级大学生创新创业训练计划项目（202010061003）；国家自然科学基金项目（31671611）；国家自然科学基金青年基金项目(31801446)；天津市研究生科研创新项目（2020YJSS131） |
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| 中文摘要: |
| 为获取高质量的大麦幼苗胚芽鞘保卫细胞以进行转录组相关研究，以大麦品种Morex的幼苗胚芽鞘为实验材料，分别对胚芽鞘表皮条撕取方法和保卫细胞分离方法进行优化。结果表明，通过“刮压”技术，不仅可以获得完整的大麦胚芽鞘表皮条，而且缩短了获取的时间；利用纤维素酶R 10与离析酶R 10制备酶解液，同时在酶解液中加入转录抑制剂，并结合表皮条“穿孔”进行辅助酶解，发现酶解温度为30 ℃、酶解时间为水浴1.5 h时，可以获取大量具有活性的大麦胚芽鞘保卫细胞。 |
| 英文摘要: |
| In order to obtain high-quality coleoptile guard cells of barley seedlings for transcriptome analysis, the methods of epidermal strip tearing and guard cell isolation from coleoptiles of barley variety Morex were optimized. The results showed that “scraping and pressing” technology can obtain the intact barley coleoptile surface strips, and shortened the time. The one-step enzymatic hydrolysis with cellulase R-10,segregation enzyme R-10,and transcriptase inhibitor in the enzymatic hydrolysate, combined with the “puncture” epidermal strip for the auxiliary enzymatic hydrolysis was performed. The optimal temperature and time of enzymatic hydrolysis for removing the epidermal cells was 30 ℃ and 1.5 h in water bath. A large number of active barley coleoptile guard cells were obtained. |
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