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Development and validation of a PCR-based functional marker system for the brown planthopper resistance gene Bph14 in rice
Authors:Lei Zhou  Zhijun Chen  Xuyong Lang  Bo Du  Kai Liu  Guocai Yang  Gang Hu  Sanhe Li  Guangcun He  Aiqing You
Institution:1.Hubei Key Laboratory of Food Crop Germplasm and Genetic Improvement, Food Crops Institute, Hubei Academy of Agricultural Sciences, Wuhan 430064, China;2.College of Life Sciences, Wuhan University, Wuhan 430072, China
Abstract:Brown planthopper (BPH) is the most damaging rice pest globally. Resistant varieties are the most effective and environmental strategy for protecting the rice crop from BPH. Functional markers (FMs) designed from polymorphic sites within gene sequences affecting phenotypic variation are highly efficient when used for marker assisted selection (MAS). Bph14 is the first and only cloned insect resistance gene so far in rice. Compared to the sequences of its non-effective alleles there are a number SNP differences. In this study, the method of allele-specific amplification (ASA) was adopted to design a simple, co-dominant, functional marker Bph14P/N for Bph14. Bph14P/N was combined with two specific dominant markers: one, named Bph14P, targets the promoter region of Bph14 and amplifies 566 bp fragments; and the other, Bph14N, targets the LRR region of bph14 and amplifies 345 bp fragments. Specificity and applicability of the functional marker system were verified in two breeding populations and a Chinese mini core collection of Oryza sativa. We recommend the use of this simple, low-cost marker system in routine genotyping for Bph14 in breeding populations.
Keywords:Oryza sativa  BPH resistance  allele-specific amplification (ASA)  MAS  functional marker
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