Chen Meiyan, He Xinhua, Huang Xing, Lu Tingting, Zhang Yili, Zhu Jiawei, Yu Haixia, Luo Cong
State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, National Demonstration Center for Experimental Plant Science Education, College of Agriculture, Guangxi University, Nanning, 530004 Guangxi People's Republic of China.
Physiol Mol Biol Plants. 2022 Jul;28(7):1407-1419. doi: 10.1007/s12298-022-01212-5. Epub 2022 Aug 2.
In this study, we selected eight cis-elements: AAAG, ACGTG, CCGA, ACTCAT, GGTCA, TATCC, TGAC and GATAA, which are closely related to plant growth and development, signal transduction and stress response. The CEAP primers were 18 nucleotides long and consisted of a central cis-element nucleotide core flanked by a filler sequence at the 5' end and di- or tri-nucleotides at the 3' end. A total of two hundred and twenty-four primers were developed, and the PCR procedure consisted of 5 cycles of low-temperature annealing and 35 subsequent cycles of annealing at 50°C. The PCR products are electrophoretically separated by 1.8-2.3% agarose. The polymorphism of the CEAP marker was amplified in eight mango ( L.) species. The results showed that the CEAP primers could amplify clear, repeatable bands in mango and combine at least four cis-elements from which a large number of bands were amplified and six highly polymorphic primers for each cis-element can reach an accurate clustering result. The results of CEAP marker assays compared with ISSR, CBDP and iPBS marker assays showed that CEAP marker was better than the other three markers in the number of fragment bands, H and I indexes. In addition, we also tested the CEAP markers in rice, tomato, potato, wax gourd, citrus and longan and the results showed that the CEAP marker assay could amplify clear polymorphic bands in different species. Our results indicate that the CEAP markers could be universally used in different species for genetic diversity analysis, relationship analysis, and marker-assisted selection for breeding.
The online version contains supplementary material available at 10.1007/s12298-022-01212-5.
在本研究中，我们选择了八个顺式作用元件：AAAG、ACGTG、CCGA、ACTCAT、GGTCA、TATCC、TGAC和GATAA，它们与植物生长发育、信号转导和应激反应密切相关。CEAP引物长度为18个核苷酸，由中央顺式作用元件核苷酸核心组成，其5'端侧翼为填充序列，3'端为二核苷酸或三核苷酸。共开发了224种引物，PCR程序包括5个低温退火循环和随后35个50℃退火循环。PCR产物通过1.8 - 2.3%的琼脂糖进行电泳分离。CEAP标记的多态性在八个芒果（L.）品种中得到扩增。结果表明，CEAP引物可在芒果中扩增出清晰、可重复的条带，并结合至少四个顺式作用元件，从中扩增出大量条带，每个顺式作用元件的六个高度多态性引物可得到准确的聚类结果。CEAP标记分析结果与ISSR、CBDP和iPBS标记分析结果相比，在片段带数量、H和I指数方面，CEAP标记优于其他三种标记。此外，我们还在水稻、番茄、马铃薯、冬瓜、柑橘和龙眼上测试了CEAP标记，结果表明CEAP标记分析可在不同物种中扩增出清晰的多态性条带。我们的结果表明，CEAP标记可普遍用于不同物种的遗传多样性分析、亲缘关系分析和育种的标记辅助选择。
在线版本包含可在10.1007/s12298-022-01212-5获取的补充材料。
