Combined expression of chitinase and β-1,3-glucanase genes in indica rice ( Oryza sativa L.) enhances resistance against Rhizoctonia solani

Abstract

Agrobacterium-mediated transformation of rice was done using the binary vector pNSP3, harbouring the rice chitinase ( chi11) gene under maize ubiquitin promoter and the tobacco β-1,3-glucanase gene under CaMV 35S promoter in the same T-DNA. Four of the six T 0 plants had single copies of complete T-DNAs, while the other two had complex integration patterns. Three of the four single-copy lines showed a 3:1 segregation ratio in the T 1 generation. Northern and western blot analyses of T 1 plants revealed constitutive expression of chitinase and β-1,3-glucanase genes. Homozygous T 2 plants of the single-copy lines CG20, CG27 and CG53 showed 62-, 9.6- and 11-fold higher chitinase activity over the control plants. β-1,3-Glucanase activity was 1.1- to 2.5-fold higher in the transgenic plants. Bioassay of homozygous T 2 plants of the three single-copy transgenic lines against Rhizoctonia solani revealed a 60% reduction in sheath blight Disease Index in the first week. The Disease Index increased from 61.8 in the first week to 90.6 in the third week in control plants, while it remained low (26.8–34.2) in the transgenic T 3 plants in the corresponding period, reflecting the persistence of sheath blight resistance for a longer period.