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Optimization of RNA In Situ Hybridization for mRNA Localization Detection in Mature Tissue of Cucumber Seedlings

Beijing Key Laboratory of Growth and Developmental Regulation for Protected Vegetable Crops, China Agricultural University, Beijing 100193, China
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Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Plants 2020, 9(11), 1461; https://doi.org/10.3390/plants9111461
Received: 7 October 2020 / Revised: 23 October 2020 / Accepted: 27 October 2020 / Published: 29 October 2020
Cucumber (Cucumis sativus L.) is one of the main vegetable crops in China. The physiological cultivation mechanism and gene function characteristics of cucumber are of great significance to the construction of modern agriculture. Due to the low genetic transformation rate of cucumber, only in situ hybridization, which does not involve the progress of gene modified transformation, is convenient to study mRNA localization, so it is more suitable for determination on mRNA localization in the mature tissue of cucumber. At present, the existing in situ hybridization technology system is more suitable for cucumber meristem than for the mature tissue of cucumber seedlings. Therefore, we optimized the traditional plant in situ hybridization protocol. Taking a known gene CsNPF7.2 (Nitrate Transporter Families protein) as an example, we then optimized the steps of plant tissue culture, gene probe preparation, plant material sampling and fixation, preparation of cross section, hybridization pretreatment, hybridization incubation, chromogenic reaction, microscopy examination, and treatment after reaction termination in order to obtain a new RNA in situ hybridization technique suitable for identification on mRNA localization in mature tissues of cucumber seedlings. This optimized technique will ensure the yield of probes, the integrity of RNA molecules, and the clarity and integrity of plant tissue structure, which is conducive to the study of gene function and screening of key genes in cucumber. View Full-Text
Keywords: cucumber; mature tissue of seedling; RNA in situ hybridization; mRNA localization cucumber; mature tissue of seedling; RNA in situ hybridization; mRNA localization
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MDPI and ACS Style

Liu, Z.; Hu, X.; Nie, J.; Li, X.; Wang, Q.; Liu, W.; Wang, T.; Lu, X.; Gao, S.; Gao, L.; Zhang, W. Optimization of RNA In Situ Hybridization for mRNA Localization Detection in Mature Tissue of Cucumber Seedlings. Plants 2020, 9, 1461. https://doi.org/10.3390/plants9111461

AMA Style

Liu Z, Hu X, Nie J, Li X, Wang Q, Liu W, Wang T, Lu X, Gao S, Gao L, Zhang W. Optimization of RNA In Situ Hybridization for mRNA Localization Detection in Mature Tissue of Cucumber Seedlings. Plants. 2020; 9(11):1461. https://doi.org/10.3390/plants9111461

Chicago/Turabian Style

Liu, Zixi, Xi Hu, Jing Nie, Xiaojun Li, Qing Wang, Wenqian Liu, Tao Wang, Xiaohong Lu, Shunli Gao, Lihong Gao, and Wenna Zhang. 2020. "Optimization of RNA In Situ Hybridization for mRNA Localization Detection in Mature Tissue of Cucumber Seedlings" Plants 9, no. 11: 1461. https://doi.org/10.3390/plants9111461

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