Research Article

Rapid Detection of Rice Fragrance Allele badh2-E7 by Recombinant Polymerase Amplification (RPA)  

Jihua Zhou1* , Anpeng Zhang1* , Wenke Xu2 , Can Cheng1 , Fuan Niu1 , Bin Sun1 , Liming Cao1 , Jianming Zhang1 , Huangwei Chu1
1 Institute of Crop Breeding and Cultivation, Shanghai Academy of Agricultural Sciences, Shanghai, 201403, China
2 Shanghai Chongming Sanxing Town Agricultural Comprehensive Technology Extension Service Center, Shanghai, 202152, China
* These authors contributed equally to this work
Author    Correspondence author
Bioscience Methods, 2024, Vol. 15, No. 1   doi: 10.5376/bm.2024.15.0001
Received: 23 Jan., 2024    Accepted: 26 Jan., 2024    Published: 31 Jan., 2024
© 2024 BioPublisher Publishing Platform
This article was first published in Molecular Plant Breeding in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Zhou J.H., Zhang A.P., Xu W.K., Cheng C., Niu F.A., Sun B., Cao L.M., Zhang J.M., and Chu H.W., 2024, Rapid detection of rice fragrance allele badh2-E7 by recombinant polymerase amplification (RPA), Bioscience Method, 15(1): 1-7 (doi: 10.5376/bm.2024.15.0002)

Abstract

Fragrant rice is favored deeply by consumers due to the strong fragrance. Fragrant rice is mainly caused by the loss-of-function mutation of the Betaine aldehyde dehydrogenase 2 (Badh2) gene in rice. Badh2-E7, with 8 bp deletion and 3 bp substitution in exon 7 of Badh2, is the main mutation allele used in fragrant rice breeding. In this study, a genotyping method named RPA-badh2-E7 for Badh2-E7 allele was designed. This method has the characteristics of rapid (completed amplification in 5 min), sensitivity (100-fold than conventional PCR), no strict amplification conditions required (25 °C~45 °C), and independent of PCR amplifier (only one thermostatic incubator is enough for amplification). This method greatly improved the efficiency of molecular marker-assisted selection of rice fragrant genes and breeding of fragrant rice varieties.

Keywords
Rice; Fragrant gene; badh2-E7; RPA
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