Brassica rapa cysteine protease 3 (BrCP3) is an endopeptidase member of C1A family (papain-like) and CA clan of cysteine protease. Members of papain-like cysteine protease (CP) have emerged to be key enzymes involved in cell death in response to biotic and environmental stresses. Recent progress of CP research features its diverse and overlapping role in basal and effector-triggered immunity. This study was carried out to characterize BrCP3 gene in Chinese cabbage and explore its function in rice – Xanthomonas oryzae pv. oryzae pathosystem. Phylogenetic analysis and BLASTP search of its amino acid sequence to the Arabidopsis Protein TAIR 10 database identified BrCP3 as the closest ortholog of Arabidopsis RD21a. The full-length cDNA of BrCP3 was cloned and transformed into the genome of japonica rice cv. ‘Gopum’. Constitutive overexpression of the transgene is driven by Ubi-1 promoter in pFLCIII vector. Regenerated single copy overexpression rice lines were inoculated with three Xoo Korean isolates. Regulation of rice resistance through cysteine protease activity is evident in the overexpression lines which showed enhanced immunity to three isolates as depicted by reduced lesion length compared with the wild type. Our initial results implicate an interesting biological function of BrCP3 in rice system that has never been reported before.

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An increasing preference for good eating quality of rice among consumers has become one of the important considerations in rice breeding. Amylose content is a leading factor affecting eating quality of rice. Amylose composition is determined by the relative activity of soluble starch synthase (SSS) and granule-bound starch synthase (GBSS). This study focused on modifying the expression of SSSI gene which is responsible for amylopectin and amylose synthesis in rice by using RNA interference (RNAi) technology. The transgenic rice plants showed various amylose contents (11–17%) in rice grains. Favorable rice lines were selected according to genomic PCR, transgene expression and amylose contents analysis. A semi-quantitative RT-PCR was carried out to determine the expression level of SSSI gene after flowering of transgenic rice and wild type. Down-regulation of SSSI gene in transgenic plants was evident in the decreasing expression in rice grains. Accordingly, scanning electron microscopy (SEM) analysis revealed uniform size with smooth curves starch granules in down-regulation rice lines, in contrast with the non-uniform granules in wild type. Results indicated that RNAi-SSSI transgenic lines produced low amylose contents that fell between glutinous and non-glutinous rice. This study showed that down-regulation of endogenous SSSI may improve the eating quality in rice.

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