Bulk segregant analysis identifies SSR markers associated with Alternaria blight resistance in linseed (Linum usitatissimum L.)

ABSTRACT

Aim: Alternaria blight caused by Alternaria lini is one of the major diseases of linseed which severely affects the yield and productivity. Here, we utilizes F2 mapping population derived from a resistant (JRF-4) and a susceptible (Chambal) genotypes of linseed and SSRs to identify the markers associated with Alternaria blight resistance using bulk segregant analysis approach. Methodology: A population consisting of 154 F2 individuals was developed from the cross between JRF-4 (resistant) and Chambal (susceptible). All 154 F2 individuals were screened with 100 polymorphic SSRs to identify extreme phenotype. Two bulk of extremes phenotypes (disease resistant and disease susceptible) from F2 mapping population were used for the bulked segregant analysis. The SSR primers that distinguished the parental lines were used to amplify the DNA from two bulks and banding pattern was observed to identify the SSRs that can differentiate the resistant and susceptible phenotypes bulk for Alternaria blight. Markers validation was carried out by amplifying DNA from individual plants of each bulk. Results: Out of 100, only 10 markers showed polymorphism among the bulks and of which only three markers viz., LUSc 898_3_12, Lu 2472 and Lu 3078 were able to differentiate the disease resistant and susceptible individuals from F2 population. Further, single marker linear regression approach was used to validate the association of selected polymorphic markers with the disease. The markers LUSc 898_3_12 and Lu 2472 showed significant regression which confirmed their linkage with Alternaria blight resistance. Interpretation: The two markers having significant regression can be used for diseases resistance breeding during marker assisted selection.