Multiplex PCR genotyping for five bacterial blight resistance genes applied to marker-assisted selection in rice (Oryza sativa)
Journal
Plant Breeding
Journal Volume
135
Journal Issue
3
Pages
309-317
Date Issued
2016-06
Author(s)
Ruishen Yap
Yu-Chia Hsu
Yong-Pei Wu
Yann-Rong Lin
Chei-Wei Kuo
Abstract
Bacterial blight (BB) is the most economically damaging disease of rice in Asia and other parts of the world. In this study, a multiplex PCR genotyping method was developed to simultaneously identify genotypes of five BB resistance genes, Xa4, xa5, Xa7, xa13 and Xa21. The resistance R alleles were amplified using five functional markers (FMs) to generate amplicons of 217, 103, 179, 381 and 595bp in IRBB66. Amplicons of 198, 107, 87, 391 and 467bp corresponded to susceptible alleles in Taiwanese japonica rice cultivars. In backcross breeding programmes, the multiplex PCR assay was integrated into selection from a population using BB resistance donor IRBB66 crossed to rice cultivar Tainung82'. Two plants with homozygosity for Xa4, xa5, Xa7, xa13 and Xa21 were selected from 1100 BC2F2 plants. In addition, the five BB resistance genes were also accurately identified in F-2 populations. This multiplex PCR method provides a rapid and efficient method for detecting various BB resistance genes, which will assist in pyramiding genes to improve durability of BB resistance in Taiwanese elite rice cultivars.
Subjects
Oryza sativa
multiplex PCR genotyping
bacterial blight resistance
marker-assisted selection
Publisher
Wiley
Type
journal article