Abstract
BackgroundRice sheath blight, caused by Rhizoctonia solani is one of the most devastating diseases of rice. It is associated with significant reduction in rice productivity worldwide. A mutant variant of mannose binding Allium sativum leaf agglutinin (mASAL) was previously reported to exhibit strong antifungal activity against R. solani. In this study, the mASAL gene has been evaluated for its in planta antifungal activity in rice plants.ResultsmASAL was cloned into pCAMBIA1301 binary vector under the control of CaMV35S promoter. It was expressed in an elite indica rice cv. IR64 by employing Agrobacterium tumefaciens-mediated transformation. Molecular analyses of transgenic plants confirmed the presence and stable integration of mASAL gene. Immunohistofluorescence analysis of various tissue sections of plant parts clearly indicated the constitutive expression of mASAL. The segregation pattern of mASAL transgene was observed in T1 progenies in a 3:1 Mendelian ratio. The expression of mASAL was confirmed in T0 and T1 plants through western blot analysis followed by ELISA. In planta bioassay of transgenic lines against R. solani exhibited an average of 55 % reduction in sheath blight percentage disease index (PDI).ConclusionsThe present study opens up the possibility of engineering rice plants with the antifungal gene mASAL, conferring resistance to sheath blight.
Highlights
Rice sheath blight, caused by Rhizoctonia solani is one of the most devastating diseases of rice
We report the stable transformation of an elite indica rice, IR-64, with the antifungal gene mannose binding Allium sativum leaf agglutinin (mASAL)
Development of mASAL expressing transgenic plants The plant expression cassette comprising cauliflower mosaic virus 35S (CaMV35S) promoter, a 333 bp mASAL coding sequence and a nos terminator was cloned into HindIII/EcoRI site of pCAMBIA1301
Summary
Rice sheath blight, caused by Rhizoctonia solani is one of the most devastating diseases of rice. It is associated with significant reduction in rice productivity worldwide. A mutant variant of mannose binding Allium sativum leaf agglutinin (mASAL) was previously reported to exhibit strong antifungal activity against R. solani. Sheath blight is considered to be an important disease of rice next to the blast disease. It is caused by a cosmopolitan, soil-borne basidiomycete necrotrophic fungus Rhizoctonia solani Kühn (teleomorph: Thanatephorus cucumeris). Sheath blight results in severe damage and reduces the rice yield by 8–50 % in the rice-growing countries of Asia [1]. The damage caused by sheath blight has become more alarming nowadays, due to the increased use of semi-dwarf, nitrogen-responsive
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