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Transformation of potato using an antisense class I patatin gene and its effect on microtuber formation

Published online by Cambridge University Press:  12 February 2007

Si Huai-Jun ,
Liu Jun  and
Xie Cong-Hua
Si Huai-Jun
Affiliation:
College of Horticulture and Forestry, Huazhong Agricultural University, Wuhan 430070, China College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
Liu Jun
Affiliation:
College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China
Xie Cong-Hua*
Affiliation:
College of Horticulture and Forestry, Huazhong Agricultural University, Wuhan 430070, China
*
*Corresponding author: Email: [email protected]
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Abstract

An antisense class I patatin gene under control of the CaMV 35S promoter was introduced into potato (Solanum tuberosum) cultivar E-potato 3 using the Agrobacterium tumefaciens system. PCR amplification and PCR–Southern blot analysis indicated that the antisense class I patatin gene had been integrated into the potato genome. Northern hybridization analysis showed that the antisense gene transcribed normally in the transgenic potato plants and resulted in a reduction of endogenous class I patatin mRNA. Total soluble protein content and lipid acyl hydrolase activity of microtubers, derived from transformed plants, decreased by a maximum of 36.4% and 31.4%, respectively, compared with control plants. The expression of this antisense gene also resulted in reductions of the plantlets forming tubers, tubers per plantlet and the effective tubers (≥50 mg) of the transformed plants.

Keywords

potatoclass I patatin geneantisense RNAmicrotuber
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 2 , Issue 1 , April 2005 , pp. 7 - 11
Copyright
Copyright © China Agricultural University and Cambridge University Press 2005

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References

Andrews, DL, Beames, B, Summers, MD and Park, WD (1988) Characterization of the lipid acyl hydrolase activity of the major potato (Solanum tuberosum) tuber protein, patatin, by cloning and abundant expression in a baculovirus vector. Biochemistry Journal 252: 199–206.CrossRefGoogle Scholar
Bradford, MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Analytical Biochemistry 72: 248–254.CrossRefGoogle ScholarPubMed
Chomczynski, P and Sacchi, N (1987) Single step method of RNA isolation by acid guanidium thiocyanate–phenol–chloroform extraction. Analytical Biochemistry 162: 156–159.CrossRefGoogle Scholar
Dai, WL, Zhu, Y, Zhao, SY and Wang, XM (1997) A new subtype of potato class I patatin gene. Acta Genetica Sinica 24: 458–463 (in Chinese with English abstract).Google ScholarPubMed
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: 1349.CrossRefGoogle ScholarPubMed
Hofgen, R and Willmitzer, L (1992) Transgenic potato plants depleted for the major tuber protein patatin via expression of antisense RNA. Plant Science 87: 45–54.CrossRefGoogle Scholar
Huang, J, Xie, C-H and Liu, J (2001) Isolation and characterization of a cDNA of patatin, the major soluble protein of potato tuber. Vestnik Saratovskogo Gosudarstvennogo Universiteta 1: 65–70.Google Scholar
Koda, Y and Kikuta, Y (1994) Wound-induced accumulation of jasmonic acid in tissues of potato tubers. Plant Cell Physiology 35: 751–756.CrossRefGoogle Scholar
Liu, J (2001) Mechanism and regulation of microtuber formation of potato. PhD thesis, Huazhong Agricultural University, Wuhan (in Chinese with English abstract).Google Scholar
Liu, J, Xie, CH and Huang, DE (1994) Research on the forming mechanism of potato microtubers: effects of dark and day length on the formation of microtubers. Chinese Potato Journal 8: 138–141 (in Chinese with English abstract).Google Scholar
Mignery, GA, Pikaard, CS and Park, WD (1988) Molecular characterization of the patatin multigene family of potato. Gene 62: 27–44.CrossRefGoogle ScholarPubMed
Murashige, T and Skoog, F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologica Plantarum 15: 473–497.CrossRefGoogle Scholar
Park, WD, Blackwood, C, Mignery, GA, Hermodson, MA and Lister, RM (1983) Analysis of the heterogeneity of the 40,000 molecular weight tuber glycoprotein of potatoes by immunological methods and by NH 2-terminal sequence analysis. Plant Physiology 71: 156–160.CrossRefGoogle Scholar
Racusen, D (1984) Lipid acyl hydrolase of patatin. Canadian Journal of Botany 62: 1640–1644.CrossRefGoogle Scholar
Racusen, D and Foote, M (1980) A major soluble glycoprotein of potato tubers. Journal of Food Biochemistry 4: 43–52.CrossRefGoogle Scholar
Si, HJ, Liu, J and Xie, CH (2003a) Expression of a novel cDNA of potato patatin class I gene in transgenic tobacco and its lipid acyl hydrolase activity. Journal of Agricultural Biotechnology 11: 236–240 (in Chinese with English abstract).Google Scholar
Si, HJ, Xie, CH and Liu, J (2003b) An efficient protocol for Agrobacterium-mediated transformation of microtuber and the introduction of an antisense class I patatin gene into potato. Acta Agronomica Sinica 29: 801–805.Google Scholar
Sonnewald, U, Studer, D, Rocha-Sosa, M and Willmitzer, L (1989) Immunocytochemical localization of patatin, the major glycoprotein in potato (Solanum tuberosum L.) tubers. Planta 178: 176–183.CrossRefGoogle ScholarPubMed
Stiekema, WJ, Heidekamp, F, Dirkse, WG, Beckum, JV, Haan, PD, Bosch, CT et al. , (1988) Molecular cloning and analysis of four potato tuber mRNAs. Plant Molecular Biology 11: 255–269.CrossRefGoogle ScholarPubMed
Twell, D and Ooms, G (1988) Structural diversity of the patatin gene family in potato cv. Desiree. Molecular & General Genetics 212: 325–336.CrossRefGoogle ScholarPubMed
Wolters, AMA and Visser, RGF (2000) Gene silencing in potato: allelic differences and effect of ploidy. Plant Molecular Biology 43: 377–386.CrossRefGoogle ScholarPubMed