Optimization and application of real-time PCR method for detecting the expression levels of nitrogen assimilation-related genes in rice |
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| Authors: | S P Zhao X Q Zhao S M Li W M Shi |
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| Institution: | (1) Graduate School of Chinese Academy of Sciences, Beijing, 100000, China;(2) State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing, 210008, China |
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| Abstract: | By optimizing the Mg2+ concentration, Taq enzyme dosage, SYBR Green I (SGI) concentration, and plate reading temperature in PCR system, we established
the method for detecting the expression levels of nitrogen assimilation-related genes in rice by using RT-PCR technique. Based
on this qualified method, we investigated the variations of OsAMT1.1 (one of nitrogen uptake genes) and OsGlt1 (one of nitrogen metabolism genes) expression levels in rice seedlings under conditions of varying nitrogen supply. The results
show that by optimizing the parameters in the PCR system to fit the characters of target genes best, we can successfully quantify
the low-abundant nitrogen transport and metabolism genes in rice quickly and exactly using fluorescence RT-PCR technique.
Published in Russian in Fiziologiya Rastenii, 2006, Vol. 53, No. 4, pp. 625–636.
The text was submitted by the authors in English. |
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| Keywords: | Oryza sativa gene expression PCR optimization real-time PCR nitrogen assimilation |
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