2013Chinese Science BulletinOpen access

Genetic analysis and gene cloning of a triangular hull 1 (tri1) mutant in rice (Oryza sativa L.)

Gongneng Feng, Changquan Zhang, Mingyong Tang, Guiyun Zhang, Chenwu Xu, Minghong Gu, Qiaoquan Liu

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Abstract

Grain shape and size are two key factors that determine rice yield and quality. In the present study, a rice triangular hull mutant ( tri1 ) was obtained from the progeny of japonica rice variety Taipei 309 treated with 60 Co γ -rays. Compared to the wild type, the tri1 mutant presents a triangular hull, and exhibits an increase in grain thickness and protein content, but with a slight decrease in plant height and grain weight. Genetic analysis indicated that the mutant phenotype was controlled by a recessive nuclear gene which is stably inherited. Using a map-based cloning strategy, we fine-mapped tri1 to a 47-kb region between the molecular markers CHR0122 and CHR0127 on the long arm of chromosome 1, and showed that it co-segregates with the molecular marker CHR0119. According to the rice genome sequence annotation there are six predicated genes within the mapped region. Sequencing analysis of the mutant and the wild type indicated that there was a deletion of an A nucleotide in exon 3 of the OsMADS32 gene, which could result in a downstream frameshift mutation and premature termination of the predicted polypeptide. Both semi-quantitative and real-time RT-PCR analyses showed that this gene expressed highly in young inflorescences, while expressed at very low levels in other tissues. These results implied that the OsMADS32 gene could be a candidate of TRI1 . Taken together, the results of this study lay the foundation for further investigation into the molecular mechanisms regulating rice caryopsis development.

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Grain shape and size are two key factors that determine rice yield and quality. In the present study, a rice triangular hull mutant ( tri1 ) was obtained from the progeny of japonica rice variety Taipei 309 treated with 60 Co γ -rays. Compared to the wild type, the tri1 mutant presents a triangular hull, and exhibits an increase in grain thickness and protein content, but with a slight decrease in plant height and grain weight. Genetic analysis indicated that the mutant phenotype was controlled by a recessive nuclear gene which is stably inherited. Using a map-based cloning strategy, we fine-mapped tri1 to a 47-kb region between the molecular markers CHR0122 and CHR0127 on the long arm of chromosome 1, and showed that it co-segregates with the molecular marker CHR0119. According to the rice genome sequence annotation there are six predicated genes within the mapped region. Sequencing analysis of the mutant and the wild type indicated that there was a deletion of an A nucleotide in exon 3 of the OsMADS32 gene, which could result in a downstream frameshift mutation and premature termination of the predicted polypeptide. Both semi-quantitative and real-time RT-PCR analyses showed that this gene expressed highly in young inflorescences, while expressed at very low levels in other tissues. These results implied that the OsMADS32 gene could be a candidate of TRI1 . Taken together, the results of this study lay the foundation for further investigation into the molecular mechanisms regulating rice caryopsis development.

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Available abstract

Grain shape and size are two key factors that determine rice yield and quality. In the present study, a rice triangular hull mutant ( tri1 ) was obtained from the progeny of japonica rice variety Taipei 309 treated with 60 Co γ -rays. Compared to the wild type, the tri1 mutant presents a triangular hull, and exhibits an increase in grain thickness and protein content, but with a slight decrease in plant height and grain weight. Genetic analysis indicated that the mutant phenotype was controlled by a recessive nuclear gene which is stably inherited. Using a map-based cloning strategy, we fine-mapped tri1 to a 47-kb region between the molecular markers CHR0122 and CHR0127 on the long arm of chromosome 1, and showed that it co-segregates with the molecular marker CHR0119. According to the rice genome sequence annotation there are six predicated genes within the mapped region. Sequencing analysis of the mutant and the wild type indicated that there was a deletion of an A nucleotide in exon 3 of the OsMADS32 gene, which could result in a downstream frameshift mutation and premature termination of the predicted polypeptide. Both semi-quantitative and real-time RT-PCR analyses showed that this gene expressed highly in young inflorescences, while expressed at very low levels in other tissues. These results implied that the OsMADS32 gene could be a candidate of TRI1 . Taken together, the results of this study lay the foundation for further investigation into the molecular mechanisms regulating rice caryopsis development.

Key concepts: Mutant, Biology, Gene, Genetics, Oryza sativa, Genetic analysis, Cloning (programming), Candidate gene

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