Development and validation of a PCR-based functional marker system for the brown planthopper resistance gene Bphl4 in rice

Lei Zhou; Zhijun Chen; Xuyong Lang; Bo Du; Kai Liu; Guocai Yang; Gang Hu; Sanhe Li; Guangcun He; Aiqing You
September 2013
Breeding Science;Sep2013, Vol. 63 Issue 3, p347
Academic Journal
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). Bphl4 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 mark-er Bphl4P/N for Bphl4. Bphl4P/N was combined with two specific dominant markers: one, named Bphl4P, targets the promoter region of Bphl4 and amplifies 566 bp fragments; and the other, Bphl4N, targets the LRR region of bphl4 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 Bphl4 in breeding populations.


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