Abstract:In order to speed up the creation of cytoplasmic male sterility (CMS) restorer line with phytophthora blight resistance in processing type pepper, a hybrid combination was prepared with long fruit inbred line 481410 as female parent and disease resistant restorer lines 9391 and 1021 (1) 1 as male parent. Rf gene and disease resistant gene were introduced into long fruit inbred lines by anther culture technology, molecular marker assisted selection (MAS) was used to preliminarily determine whether DH lines contain Rf gene and disease resistance. The disease resistance of DH lines was verified by seedling resistance identification. The results showed that: (1) twentytwo embryoids were induced by donor parent (481410×9391) F1, and eleven DH lines were obtained. 9 DH lines were obtained from donor [481410×1021(1)1] F1. (2) The PCR results of molecular marker CRFSCAR showed that seven DH lines obtained from donor (481410×9391) F1 could amplify 870 bp specific bands, accounting for 63.6%. However, eight DH lines from donor [481410×1021(1)1] F1 could also amplify 870 bp specific bands, accounting for 88.9%. (3) PCR results of FQ01/RQ01 molecular marker showed that five DH lines from donor (481410×9391) F1 could amplify 717 bp specific bands, accounting for 45.5%. Four DH lines from donor [481410×1021(1)1] F1 could amplify specific bands of 717 bp, accounting for 44.4%. (4) Seven phytophthora blight resistance DH lines carrying Rf gene were preliminarily screened by MAS technology. The identification results of seedling resistance showed that five DH lines were disease resistant and two were moderately disease resistant among the seven DH lines. The investigation of agronomic characters and the test crossing experiment showed that ‘Yulaxuan 32’ and ‘Yulaxuan 71’were single pepper with longer fruit and spicy taste, and both were cytoplasmic male sterility (CMS) restorers. The two DH lines created in this study will lay a foundation for breeding new disease resistant processing pepper varieties with CMS three lines.