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Phenotypic and Candidate Gene Analysis of a New Floury Endosperm Mutant (osagpl2-3) in Rice

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Abstract

A floury endosperm mutant, osagpl2-3, was isolated from the M2 generation of japonica rice cultivar Nipponbare following ethyl methane sulfonate mutagenesis. The osagpl2-3 mutant produced a white-core endosperm compared to the transparent endosperm of the wild type (WT). The results from scanning electron microscope showed that the osagpl2-3 mutant grains comprised of round and loosely packed starch granules, some of which were compounded. The analysis for cooking and nutrition quality traits indicated that the values of gel consistency, gelatinization temperature, and rapid viscosity analysis profile of osagpl2-3 grains were lower than those of the WT. Besides, the protein content, the contents of nine different amino acids, and the thermodynamic parameters of T p and ΔT 1/2 in osagpl2-3 were also different from those of the WT. Genetic analysis revealed that osagpl2-3 mutation was controlled by a single recessive gene. The osagpl2-3 gene was mapped between InDel markers R1M30 and ID1-12 on rice chromosome 1. In the candidate region of the Nipponbare genome, an annotated gene, LOC_Os01g44220 which encodes a large subunit of putative ADP-glucose pyrophosphrylase named OsAPL2 was considered the optimal candidate. Cloning and sequencing of LOC_Os01g44220 in different plants of the osagpl2-3 mutants revealed a single nucleotide mutation (G→A) in the open reading frame region, which led to a substitution of an acidic amino acid Glu (E) by a basic amino acid Lys (K) accordingly. Furthermore, the mutant site is close to the functional domain which interacts with the ADP-Glc. In brief, these results suggested that the osagpl2-3 is a new mutant of OsAPL2.

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Abbreviations

AC:

Amylose content

ADPG:

ADP-glucose

AGPase:

ADP-glucose pyrophosphorylase

ATP:

Adenosine 5′ triphosphate

GC:

Gel consistency

Glc-1-P:

Glucose-1-phosphate

GT:

Gelatinization temperature

ORF:

Open reading frame

PC:

Protein content

RVA:

Rapid viscosity analysis

PPDK:

Pyruvate orthophosphate dikinase

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Acknowledgments

This work was supported by Zhejiang Provincial Natural Science Foundation of China (no. Y3080217), the Science and Technology Office of Zhejiang Province (no. 2010C32002 and no. 2007C12902), the Program for Innovative Research Team in University(IRT1185), the Fundamental Research Funds for the Central Universities" after 2007C12902), and 151 Foundation for Talents of Zhejiang Province of China. We thank Mr. Bin Zhang and Kemin Wang for the assistance of getting some data during the project. We also thank Dr. Wenqiang Li and Alfred Quampah in the English revision of the manuscript.

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Authors and Affiliations

  1. Agronomy Department, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, 310058, China

    Dapeng Zhang, Jianguo Wu, Yujiang Zhang & Chunhai Shi

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  1. Dapeng Zhang
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  2. Jianguo Wu
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  3. Yujiang Zhang
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Correspondence to Chunhai Shi.

Additional information

Dapeng Zhang and Jianguo Wu contributed equally to this paper.

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Supplementary Table 1

Sequencing primers of candidate gene LOC_Os01g44220. (DOCX 24 kb)

Supplementary Table 2

Putative genes in candidate region. (DOCX 31 kb)

Supplementary Table 3

Physical-chemical parameters of protein transcript of LOC_Os01g44220 of WT and flo6. (DOCX 24 kb)

Supplementary Table 4

Secondary structure parameter of protein transcript of LOC_Os01g44220 of WT and flo6. (DOCX 24 kb)

Supplementary Fig. 1

Tertiary structures of protein transcript of LOC_Os01g44220 of WT and flo6 a and b Prediction tertiary of LOC_Os01g44220.1~6 from WT (a) and flo6 (b). c and d Prediction tertiary of LOC_Os01g44220.7 from WT (c) and flo6 (d). White arrows indicated the changes of secondary structure of protein between WT and flo6. (DOCX 604 kb)

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Zhang, D., Wu, J., Zhang, Y. et al. Phenotypic and Candidate Gene Analysis of a New Floury Endosperm Mutant (osagpl2-3) in Rice. Plant Mol Biol Rep 30, 1303–1312 (2012). https://doi.org/10.1007/s11105-012-0435-5

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