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Improving freezing tolerance of transgenic tobacco expressing sucrose: sucrose 1-fructosyltransferase gene from Lactuca sativa

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Abstract

Sucrose: sucrose 1-fructosyltransferase (1-SST) cDNA from Lactuca sativa, coding the enzyme responsible for lower degree polymers fructan biosynthesis, was cloned by RT-PCR and RACE methods. The 1-SST cDNA under the control of CaMV 35S promoter was introduced into tobacco by Agrobacterium-mediated leaf disc transformation protocol. Fructan synthesis in vitro and carbohydrate analysis showed that sense transgenic tobacco plant displayed sucrose: sucrose 1-fructosyltransferse activity. After freezing stress, significant increases in electrolyte leakage and malondialdehyde were found in the wild type and anti-sense transgenic plants, while no apparent differences were observed in sense transgenic plants. Meanwhile, water soluble carbohydrate, fructan and fructose of sense transgenic plants remarkably increased, compared with those of wild type and anti-sense plants. No significant difference was detected in superoxide dismutase activity between transgenic and wild type plants. The above results demonstrated that the expression of 1-SST gene improved the freezing resistance of transgenic tobacco plants.

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Abbreviations

CTAB:

Cetyltrimethyl ammonium bromide

DP:

Degree polymers

1-FFT:

Fructan: fructan 1-fructosyltransferase

MDA:

Malondialdehyde

ORF:

Open reading frame

RACE:

Rapid amplification of cDNA ends

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

1-SST:

Sucrose: sucrose 1-fructosyltransferase

TLC:

Thin layer chromatography

WSC:

Water soluble carbohydrate

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Acknowledgment

This work was supported by the National High Technology Research and Development Program of China (863 Program) (No. 2006AA10A107).

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Authors and Affiliations

  1. College of Life Science and Technology, Ocean University of China, Qingdao, 266003, People’s Republic of China

    Hui-Juan Li & Xue-Cheng Zhang

  2. School of Life Science, Shandong University, 27 Shanda South Road, Jinan, 250100, People’s Republic of China

    Ai-Fang Yang, Feng Gao & Ju-Ren Zhang

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  1. Hui-Juan Li
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  2. Ai-Fang Yang
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  3. Xue-Cheng Zhang
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  4. Feng Gao
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  5. Ju-Ren Zhang
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Correspondence to Ju-Ren Zhang.

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Li, HJ., Yang, AF., Zhang, XC. et al. Improving freezing tolerance of transgenic tobacco expressing sucrose: sucrose 1-fructosyltransferase gene from Lactuca sativa . Plant Cell Tiss Organ Cult 89, 37–48 (2007). https://doi.org/10.1007/s11240-007-9213-8

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