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Identification of WA-Type Three-Line Hybrid Rice with Real-Time Polymerase Chain Reaction (PCR) Method.
MedLine Citation:
PMID:  22161239     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
A real-time fluorescent PCR (RTF-PCR) was developed to detect and quantify wild abortive (WA)-type three-line hybrid rice (Oryza sativa L.). The mitochondrial R(2-630) WA gene was reported to be closely related to male sterility in plants, and developed as a molecular maker to identify the cytoplasmic male sterility system of hybrid rice. First, we got the DNA sequence of R(2-630) WA gene in 17 rice species with traditional PCR. Then, a pair of specific primers (P(3), P(4)) and TaqMan fluorescence probe (P(3-14)) were designed based on the R(2-630) DNA sequence. The following RTF-PCR was performed on the 17 rice species finally. The results indicate that the probes used here are specific for three-line hybrid rice F(1) and male sterile lines. We can even identify a single hybrid seed using the probes, which confirmed that the probes can be applied to the identification and quantification of the WA-type three-line hybrid rice. In addition, the RFT-PCR system can be optimized when the annealing temperature is 60 °C and the Mg(2+) concentration is 3.5 mmol/L.
Authors:
Y Cheng; B D Gao; H Y Chen; J J Mao; A X Cao; J G Zhu; S F Zhu
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-12-8
Journal Detail:
Title:  Applied biochemistry and biotechnology     Volume:  -     ISSN:  1559-0291     ISO Abbreviation:  -     Publication Date:  2011 Dec 
Date Detail:
Created Date:  2011-12-13     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8208561     Medline TA:  Appl Biochem Biotechnol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Institute of Bast-Fiber crops, Chinese Academy of Agricultural Sciences, Xianjiahu West Road 348, Changsha, China, charles_chengyi@yahoo.com.cn.
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