Abstract
BackgroundMechanical strength is a crucial agronomic trait in rice (Oryza sativa), and brittle mutants are thought suitable materials to investigate the mechanism of cell wall formation. So far, almost all brittle mutants are recessive, and most of them are defected in multiple morphologies and/or grain yield, limiting their application in hybrid breeding and in rice straw recycling.ResultsWe identified a semi-dominant brittle mutant Brittle culm19 (Bc19) isolated from the japonica variety Nipponbare through chemical mutagenesis. The mutant showed the same apparent morphologies and grain yield to the wild type plant except for its weak mechanical strength. Its development of secondary cell wall in sclerenchyma cells was affected, along with reduced contents of cellulose, hemicellulose, lignin and sugars in culms and leaves. Positional cloning suggested that the Bc19 gene was allelic to OsCESA4, encoding one of the cellulose synthase A (CESA) catalytic subunits. In this mutant, a C-to-T substitution occurred in the coding sequence of BC19, causing the P507S missense mutation in its encoded product, which was located in the second cytoplasmic region of the OsCESA4 protein. Furthermore, introducing mutant gene Bc19 into the wild-type plant resulted in brittle plants, confirming that the P507S point mutation in OsCESA4 protein was responsible for the semi-dominant brittle phenotype of Bc19 mutant. Reverse correlation was revealed between cellulose contents and expression levels of mutant gene Bc19 among the homozygous mutant, the hybrid F1 plant, and the Bc19 overexpression transgenic plants, implying that gene Bc19 might affect cellulose synthesis in a dosage-dependent manner.ConclusionsBc19, a semi-dominant brittle mutant allele of gene OsCESA4, was identified using map-based cloning approach. The mutated protein of Bc19 possessing the P507S missense mutation behaved in a dosage-dependent semi-dominant manner. Unique brittle effect on phenotype and semi-dominant genetic quality of gene Bc19 indicated its potential application in grain-straw dual-purpose hybrid rice breeding.
Highlights
Mechanical strength is a crucial agronomic trait in rice (Oryza sativa), and brittle mutants are thought suitable materials to investigate the mechanism of cell wall formation
The decrease in cell wall components of transgenic lines TG1 and TG2 were severer than those of mutant Brittle culm19 (Bc19) (Fig. 5b, c), probably because of their higher transcript levels of mutant gene Bc19 guided by the strong promoter of Actin1 (Fig. 5d). These results suggested that the brittle phenotype of the Bc19 mutant was due to the P507S substitution of OsCESA4, and the mutant Bc19 gene blocked the synthesis of essential components of cell wall in a dominant way
NE1031, NC0259, ND8759, and ND2395 were generated by Tos17 insertion in OsCESA genes, and they were aberrant in plant height, leaf size, culm thickness, and fertility (Tanaka et al 2003). bc7 was another brittle mutant allele of OsCESA4 obtained through 60Co-γ radiation, resulting in the premature transcription of the corresponding gene (Yan et al 2007)
Summary
Mechanical strength is a crucial agronomic trait in rice (Oryza sativa), and brittle mutants are thought suitable materials to investigate the mechanism of cell wall formation. SCW generally determines the mechanical strength of cell walls, so defects in biosynthesis of cellulose, hemicellulose and/ or lignin always result in inferior mechanical index and brittle plant bodies, which in turn make these brittle culm mutants valuable materials for understanding the mechanism of SCW formation. A number of brittle mutants have been studied, and some responsible genes have been identified in Arabidopsis thaliana, rice (Oryza sativa) and other cereal crops, which to some extent revealed the mechanism regulating mechanical strength of the plant body and metabolic pathway of plant cell walls. IRX4 encoding a cinnamoyl-CoA reductase (CCR) is essential for lignin biosynthesis; FRAGILE FIBER1 (FRA1) encodes a kinesin-like protein and regulates the oriented deposition of cellulose microfibrils; and mutant of gene FRA2 is attributable to altered fiber cell elongation and expansion (Jones et al 2001; Zhong et al 2002; Burk et al 2001)
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