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Overexpression of UDP-glucose pyrophosphorylase from Larix gmelinii enhances vegetative growth in transgenic Arabidopsis thaliana

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Abstract

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A UDP-glucose pyrophosphorylase gene ( LgUGPase ) was identified from Larix gmelinii, and its function in enhancing vegetative growth and cellulose biosynthesis was confirmed by analyzing transgenic Arabidopsis thaliana overexpressed LgUGPase .

Abstract

UDP-glucose pyrophosphorylase (UGPase), an important regulatory enzyme in carbohydrate metabolism, catalyzes the reversible production of glucose 1-phosphate and the conversion of uridine triphosphate to uridine diphosphate glucose and pyrophosphate. In this study, a larch UGPase (LgUGPase) gene was isolated from Larix gmelinii. The 1,443-bp open reading frame encodes a protein of 480 amino acids with a predicted molecular weight of 53.7 kDa and shows striking sequence similarity to UGPase proteins from Pinus taeda and Picea sitchensis. Semiquantitative reverse transcription-polymerase chain reaction showed that the LgUGPase gene was expressed primarily in the larch stem in addition to its root and leaf. Southern blot analysis indicated that LgUGPase is encoded by two genes in the L. gmelinii genome. Overexpression of LgUGPase enhanced vegetative growth in transgenic Arabidopsis and increased the contents of soluble sugars and cellulose, and thickened parenchyma cell walls. These results revealed that L. gmelinii UGPase participates in sucrose/polysaccharide metabolism and cell wall biosynthesis, suggesting that LgUGPase may be a good candidate gene for improvement of fiber cell development in plants.

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Abbreviations

CTAB:

Cetyltrimethylammonium bromide

Glc-1-P:

Glucose 1-phosphate

NB:

Nucleotide binding

NPT II:

Neomycin phosphotransferase II gene

ORF:

Open reading frame

RT-PCR:

Reverse transcription-polymerase chain reaction

SuSy:

Sucrose synthase

UDPG:

Uridine diphosphate glucose

UGPase:

UDP-glucose pyrophosphorylase

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Acknowledgments

We would like to thank Professor Nakamura Kenzo in Chubu University for kindly providing pBI101-35::Gus-Hm vector. This study was supported by the National Natural Science Foundation of China (No. 31160143, No. 31060106, No. 30960030 and 31260168), Natural Science Foundation of Inner Mongolia (No. 2010BS0509 and 2009MS0505) and Chun-Hui project (No. Z2009-1-01009).

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Authors and Affiliations

  1. College of Life Sciences, Inner Mongolia University, 235 Daxuexi Road, Hohhot, 010021, China

    Ningning Li, Li Wang & Xiaofei Lin

  2. College of Forestry, Inner Mongolia Agricultural University, Hohhot, 010019, China

    Wenbo Zhang

  3. Graduate School of Science and Technology, Kumamoto University, Kurokami, Kumamoto, 860-8555, Japan

    Katsuaki Takechi & Hiroyishi Takano

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  1. Ningning Li
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  2. Li Wang
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  3. Wenbo Zhang
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  4. Katsuaki Takechi
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  5. Hiroyishi Takano
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  6. Xiaofei Lin
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Corresponding author

Correspondence to Xiaofei Lin.

Additional information

Communicated by R. Schmidt.

The nucleotide sequences reported in this paper have been submitted to the DNA data bank of Japan (DDBJ) under accession number AB830476.

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Li, N., Wang, L., Zhang, W. et al. Overexpression of UDP-glucose pyrophosphorylase from Larix gmelinii enhances vegetative growth in transgenic Arabidopsis thaliana . Plant Cell Rep 33, 779–791 (2014). https://doi.org/10.1007/s00299-013-1558-3

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