DNA marker-assisted identification of male-fertility restorers helps save considerable resources, which in turn enable increasing the pace and efficiency of breeding hybrid cultivars in crops including chilli. In this article, our objective is to demonstrate the efficiency of DNA-based markers to classify untested advanced breeding lines (ABLs) into restorers and maintainers. We used four reported fertility restoration (Rf) gene-linked Sequence Characterized Amplified Region (SCAR)-based markers and one Cleaved Amplified Polymorphic Sequence (CAPS)-based marker to screen 18 ABLs for fertility restoration and sterility maintenance. One of these four markers (CRF3S1S) was successful in detecting 14 out of 18 ABLs as restorers. Other four markers namely, CRF-SCAR, CaRf-FL-M2, BAC13T7 and OPP-13-CAPS identified 12, 16, 11 and 10 restorers, respectively. We validated these DNA markers using phenotype-based male fertility indicator field and laboratory assayable traits. For this purpose, we evaluated 126 F-1 hybrids involving 18 ABLs and seven cytoplasmic-nuclear genetic male sterile lines for four field assayable male-fertility indicator traits-anther structure, pollen release score, fruit set, seed set and two laboratory assayable indicators-pollen fertility and pollen viability. Results of field and laboratory assays confirmed male fertility restoring ability of 14 out of 18 ABLs which were classified as complete restorers based on marker assay. The SCAR marker, CRF3S1S was highly efficient (100%) in differentiating restorers from maintainers. Based on these results, we suggest the use of CRF3S1S for unambiguous detection of restorers from untested ABLs and/or any germplasm at a much quicker time and limited resources.
