Marker-Assisted Pyramiding of Blast-Resistance Genes in a japonica Elite Rice Cultivar through Forward and Background Selection

Abstract

Rice blast, caused by Pyricularia oryzae, is one of the main rice diseases worldwide. The pyramiding of blast-resistance (Pi) genes, coupled to Marker-Assisted BackCrossing (MABC), provides broad-spectrum and potentially durable resistance while limiting the donor genome in the background of an elite cultivar. In this work, MABC coupled to foreground and background selections based on KASP marker assays has been applied to introgress four Pi genes (Piz, Pib, Pita, and Pik) in a renowned japonica Italian rice variety, highly susceptible to blast. Molecular analyses on the backcross (BC) lines highlighted the presence of an additional blast-resistance gene, the Pita-linked Pita2/Ptr gene, therefore increasing the number of blast-resistance introgressed genes to five. The recurrent genome was recovered up to 95.65%. Several lines carrying four (including Pita2) Pi genes with high recovery percentage levels were also obtained. Phenotypic evaluations confirmed the effectiveness of the pyramided lines against multivirulent strains, which also had broad patterns of resistance in comparison to those expected based on the pyramided Pi genes. The developed blast-resistant japonica lines represent useful donors of multiple blast-resistance genes for future rice-breeding programs related to the japonica group.

Keywords: KASP marker; Pi genes; Pyricularia oryzae; backcross; breeding; molecular markers; pyramiding; rice.

Figure 1

Molecular markers developed in this work for Pik1 (a) and Pita (b). M = molecular weight marker; Ku = Kusabue (Pik original donor); S = SJKK (donor parent); Sa = Saber; Ka = Katy (Pita original donor); VN = Vialone Nano (recurrent parent). Arrows indicate 500 bp size.

Figure 2

Position of the Pi markers listed in Table 1 and Table 2 in the Pik, Pib, Pita, and Piz homolog genes on Nipponbare reference genome IRGSP-1.0 (GenBank accessions HM048900.1 (LOC_Os11g46210, Os11g0689100), AB013448.1 (LOC_Os02g57305, Os02g0818450), AF207842.1 (LOC_Os12g18360, Os12g0281300), and RAP-DB accession Os06g0286700 (LOC_Os06g17900), respectively).

Figure 3

MABC scheme for the introgression of the four Pi genes (Pib, Piz, Pik, and Pita) where SJKK acted as donor parent (DP) and Vialone Nano as recurrent parent (RP). Numbers indicate the tested plants with respect to the number of plants having the desired marker alleles.