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Identification, cloning and sequence analysis of shallot like1-Fuhui673 from a rolled leaf mutant in rice

Leaves are the important organ for photosynthesis in rice and the strong and weak photosynthesis will be directly influenced with their morphologic structure of leaves. Moderate rolling leaf could keep plants upright, improve light transmittance of population, increase efficiency of light utilization and then increase rice yield. We obtained a rolled leaf mutant, named sll1-fh673 ( shallot like1-Fuhui673 ) derived from the restorer line Fuhui673 ( Oryza sativa L. ssp. indica ) of hybrid rice, which were treated with 60 Co-γ radiation. The leaves of the sll1-fh673 rolled inward like a cylinder during the whole growth period. The characteristics of the sll1-fh673 were distinct from those of the wild-type plants, with shorter on plant height and ear length, malformed spikelets, decreased the grain number per spike, smaller grain shape, worse development of the seed. The result of genetic analysis indicated that the phenotype of rolled leaves of sll1-fh673 was controlled by a single recessive nuclear gene. Meanwhile, the sll1-fh673 gene was mapped to a 52-kb region between inDel marker Ri9-20 and Ri9-7 on the Chromosome 9 of rice with Bulked Segregant Analysis and Recessive-class Analysis methods. But there was a cloned rolled leaf gene SLL1 (LOC_Os09g23200) in the mapped regions, which might be the candidate gene of sll1-fh673 . The sequencing analysis revealed that there was a single-base substitution from C to A and five-base deletion in the first exon in sll1-fh673 comparing with the SLL1 gene, leading to frame-shift mutation after the 148 aa and dysfunction of sll1 protein. So we speculated that the sll1-fh673 gene might be allelic to SLL1 gene. These results will deepen our understanding the mechanisms of the leaf rolling and enriches the allelic resource of the SLL1 gene. We will predict binding sites of the sll1-fh673 gene and find interacting protein of down-regulation, and then carry out the molecular mechanism of the leaf rolling, such as electrophoretic mobility shift assay (EMSA), DNA Chip Technology and Yeast One Hybrid System in the future.