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Role of tobacco vacuolar invertase regulated by patatin promoter in potato tuber resistance to cold sweetening

Published online by Cambridge University Press:  15 June 2007

Cheng Shan-Han
Affiliation:
National Center of Vegetable Improvement (Central China);Key Laboratory of Horticultural Plant Biology (Huazhong Agricultural University), Ministry of Education; Potato Engineering and Technology Research Center of Hubei Province, Wuhan 430070, China
Liu Jun*
Affiliation:
National Center of Vegetable Improvement (Central China);Key Laboratory of Horticultural Plant Biology (Huazhong Agricultural University), Ministry of Education; Potato Engineering and Technology Research Center of Hubei Province, Wuhan 430070, China
Xie Cong-Hua
Affiliation:
National Center of Vegetable Improvement (Central China);Key Laboratory of Horticultural Plant Biology (Huazhong Agricultural University), Ministry of Education; Potato Engineering and Technology Research Center of Hubei Province, Wuhan 430070, China
Song Bo-Tao
Affiliation:
National Center of Vegetable Improvement (Central China);Key Laboratory of Horticultural Plant Biology (Huazhong Agricultural University), Ministry of Education; Potato Engineering and Technology Research Center of Hubei Province, Wuhan 430070, China
Li Jing-Cai
Affiliation:
National Center of Vegetable Improvement (Central China);Key Laboratory of Horticultural Plant Biology (Huazhong Agricultural University), Ministry of Education; Potato Engineering and Technology Research Center of Hubei Province, Wuhan 430070, China
*
*Corresponding author. E-mail: [email protected]

Abstract

To slow down the accumulation of reducing sugar in potato tubers exposed to low-temperature storage, an expression vector, pBICNI, including Nt-VIF (Nicotiana tabacum vacuolar inhibitor of beta fructosidase, a vacuolar invertase inhibitor from Nicotiana tabacum) gene regulated by potato tuber specific promoter class I patatin promoter (CIPP) was constructed and transformed into potato (Solanum tuberosum L.) cultivar E-potato 3 (E3). Detection by polymerase chain reaction (PCR), Northern and Southern hybridizations indicated that the full-length Nt-VIF cDNA was transformed successfully into cv. E3. After storing potato tubers of 14 transgenic lines at 4 or 20°C for 30 days, their activities of vacuolar invertase (VI) and reducing sugar (RS) content were analysed. The results showed that there were no significant differences in RS content between transgenic and untransformed (control) tubers stored at 20°C. However, RS content of transgenic lines was obviously reduced at 4°C compared to the control, from 34.0% (line B-13) to 76.8% reduction (line B-1), implying that VI activity was inhibited by Nt-VIF cDNA expression and RS content was reduced. Further analysis revealed a positive linear relationship between VI activity and RS content (VI=0.308RS+0.067), and lines B-1, B-2, B-6, B-9 and B-14 could meet the requirements of potato chips in terms of their low RS content after cold storage.

Type
Research Article
Copyright
Copyright © China Agricultural University and Cambridge University Press 2007

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Footnotes

First published in Journal of Agricultural Biotechnology 2006, 14(5):716–720

References

Chen, F and Hu, XS (2000) The effect of storage temperature on carbohydrate content and chip color of the potato tuber. Acta Horticulturae Sinica 27(3): 218219 (in Chinese with English abstract).Google Scholar
Chen, YH, Ou Yang, B, Li, HX, et al. (2005) Isolation and characterization of cucumber ACC oxidase gene and its upstream. Agricultural Sciences in China 4(1): 16.Google Scholar
Cheng, SH, Su, ZH, Xie, CH, et al. (2004) Effects of variation in activities of starch–sugar metabolic enzymes on reducing sugar accumulation and processing quality of potato tubers. Agricultural Sciences in China 3(7): 519527.Google Scholar
Edwards, K, Johnstone, C and Thompson, C (1991) A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Research 19: 13491358.Google Scholar
Greiner, S, Rausch, T, Sonnewald, U, et al. (1999) Ectopic expression of a tobacco invertase inhibitor homolog prevents cold-induced sweetening of potato tubers. Nature Biotechnology 17: 708711.Google Scholar
Iglesias, VA, Moscone, EA, Papp, I, et al. (1997) Molecular and cytogenetic analyses of stably and unstably expressed transgene loci in tobacco. Plant Cell 9: 12511264.Google ScholarPubMed
Isherwood, FA (1973) Starch–sugar interconversion in Solanum tuberosum. Phytochemistry 12: 25792591.CrossRefGoogle Scholar
Koch, K (2004) Sucrose metabolism: regulatory mechanisms and pivotal roles in sugar sensing and plant development. Current Opinion in Plant Biology 7(3): 235246.CrossRefGoogle ScholarPubMed
Lorberth, R, Ritte, G, Willmitzer, L, et al. (1998) Inhibition of a starch-granule-bound protein leads to modified starch and repression of cold sweetening. Nature Biotechnology 16: 473477.Google Scholar
Rommens, CM, Ye, J, Humara, JM, et al. (2003) Precise breeding. U.S. Patent Application No. 221, 213.Google Scholar
Sambrook, J, Fritsch, EF and Maniatis, T (1989) Molecular Cloning. A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, New York.Google Scholar
Si, HJ, Xie, CH and Liu, J (2003) An efficient protocol for Agrobacterium-mediated transformation with microtuber and the introduction of an antisense class I patatin gene into potato. Acta Agronomy Sinica 29(6): 801805 (in Chinese with English abstract).Google Scholar
Stark, DM, Timmermann, KP, Barry, GF, et al. (1992) Regulation of the amount of starch in plant tissues by ADP glucose pyrophosphorylase. Science 258: 287292.Google Scholar
Sturm, A (1999) Invertases, primary structures, functions and roles in plant development and sucrose partitioning. Plant Physiology 121: 18.Google Scholar
Zrenner, R, Schüler, K and Sonnewald, U (1996) Soluble acid invertase determines the hexose-to-sucrose ratio in cold-stored potato tubers. Planta 198: 246252.Google Scholar