Abstract
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 (Mangifera indica 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.
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Acknowledgements
This research was supported by Science and Technology Major Project of Guangxi (GXKJ-AA18118046), State Key Laboratory for Conservation and Utilization of Subtropical Agro- bioresources (SKLCUSA-a201906, SKLCUSA-c201901), the six one’ special action of “strengthening agriculture and enriching people” by science and technology Vanguard (Guangxi Agricultural Science and technology League 202204). Innovation Team of Guangxi Mango Industry Project (nycytxgxcxtd-2021-06-02) and Innovation Project of Guangxi Graduate Education (YCSW2021043).
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C.L. and X.-H.H.; conceived of the project and financed this experiment. C.L. and M.-Y.C.; designed and executed the experiments. X.H., T.-T.L., Y.-L.Z., J.-W.Z, and H.-X.Y.; helped with the experiments, and analysed the data. M.-Y.C. and C.L.; wrote the manuscript. All authors have read and approved the final manuscript.
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We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled “Cis-element amplified polymorphism (CEAP), a novel promoter- and gene-targeted molecular marker of plants”. All authors have seen the manuscript and approved the submittal to your journal. None of the material in the paper has been published or is under consideration for publication elsewhere. We thank Bao-Xiang Qin, Rong-Bai Li, Xian-Yu Wang, Ying-Yi Deng, Zheng-Guo Liu (College of agriculture, Guangxi University, Nanning, GX) for kindly providing the materials of rice, tomato, potato, wax gourd, citrus and longan, respectively.
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Chen, M., He, X., Huang, X. et al. Cis-element amplified polymorphism (CEAP), a novel promoter- and gene-targeted molecular marker of plants. Physiol Mol Biol Plants 28, 1407–1419 (2022). https://doi.org/10.1007/s12298-022-01212-5
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DOI: https://doi.org/10.1007/s12298-022-01212-5