Construction of Tissue Culture System of Onobrychis viciaefolia Scop ‘Mengnong’ Anthers

Abstract

Background: To accelerate the establishment of a genetic improvement system for Onobrychis viciaefolia Scop. by cultivating haploid plants and producing pure parental material. Methods: We endeavored to set up the Onobrychis viciaefolia Scop. anther culture system, examine the suitable anther pre-treatment method and the factors influencing medium type and hormone ratio during anther regeneration. Result: The regenerative capacity of intact anthers was preserved by 48h low temperature (4°C) medium treatment. Or the broken anthers were obtained when the regenerated grass was budding and preserved in the medium after 48 hours of low temperature treatment; callus induction was based on MS and the callus rate was 18.33% when 1.0 mg/L 2,4-D + 0.75 mg/L 6-BA + 0.8 mg/L NAA + 2.0 mg/L KT was added. With N6 as the basic medium, the addition of 2.0 mg/L 2,4-D + 0.75 mg/L 6-BA + 0.4 mg/L NAA + 0.5 mg/L KT resulted in a healing rate of 16.19%; the addition of 0.5 mg/L 6-BA + 1.0 mg/L KT to the MS medium resulted in a differentiation rate of 68%; during the construction of the anther histoponic system, browning and waterlogging of the healing tissues and vitrification of the seedlings hindered the formation of regenerated plants.