Abstract
Granule-bound starch synthase I (GBSSI) is one of the key enzymes catalyzing the formation of amylose, a linear α(1,4)D-glucan polymer, from ADP-glucose. Amylose-free transgenic sweet potato plants were produced by inhibiting sweet potato GBSSI gene expression through RNA interference. The gene construct consisting of an inverted repeat of the first exon separated by intron 1 of GBSSI driven by the CaMV 35S promoter was integrated into the sweet potato genome by Agrobacterium tumefaciens-mediated transformation. In over 70% of the regenerated transgenic plants, the expression of GBSSI was inactivated giving rise to storage roots containing amylopectin but not amylose. Electrophoresis analysis failed to detect the GBSSI protein, suggesting that gene silencing of the GBSSI gene had occurred. These results clearly demonstrate that amylose synthesis is completely inhibited in storage roots of sweet potato plants by the constitutive production of the double-stranded RNA of GBSSI fragments. We conclude that RNA interference is an effective method for inhibiting gene expression in the starch metabolic pathway.
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References
Chang S, Puryear J, Cairney J (2000) A simple and efficient method for isolation RNA from pine trees. Plant Mol Biol Rep 11:113–116
Hamada T, Kodama H, Nishimura M, Iba K (1996) Modification of fatty acid composition by over- and antisense-expression of a microsomal ω-3 fatty acid desaturase gene in transgenic tobacco. Transgenic Res 5:115–121
Heeres P, Schippers-Rozenboom M, Jacobsen E, Visser RGF (2002) Transformation of a large number of potato varieties: genotype-dependent variation in efficiency and somaclonal variability. Euphytica 124:13–22
Heilersig HJB, Loonen A, Bergervoet M, Wolters AMA, Visser RGF (2006) Post-transcriptional gene silencing of GBSSI in potato: effect of size and sequence of the inverted repeats. Plant Mol Biol 60:647–662
Kim SH, Mizuno K, Fujimura T (2002) Isolation of MADS-box genes from sweet potato (Ipomea batatas (L.) Lam.) expressed specifically in vegetative tissues. Plant Cell Physiol 43:314–322
Kimura T, Ideta O, Saito A (2000) Identification of the gene encoding granule-bound starch synthase I in sweet potato (Ipomoea batatas (L.) Lam.). Plant Biotechnol 17:247–252
Kimura T, Otani M, Noda T, Ideta O, Shimada T, Saito A (2001) Absence of amylose in sweet potato (Ipomoea batatas (L.) Lam.) following the introduction of granule-bound starch synthase I cDNA. Plant Cell Rep 20:663–666
Kuipers AGJ, Jacobsen E, Visser RGF (1994a) Formation and deposition of amylase in the potato tuber starch granule are affected by the reduction of granule-bound starch synthase gene expression. Plant Cell 6:43–52
Kuipers AGJ, Soppe WJJ, Jacobsen E, Visser RGF (1994b) Field evaluation of transgenic potato plants expressing an antisense granule-bound starch synthase gene: increase of the antisense effect during tuber growth. Plant Mol Biol 26:1759–1773
Kuipers AGJ, Soppe WJJ, JacobsenE, Visser RGF (1995) Factors affecting the inhibition by antisense RNA of granule-bound starch synthase gene expression in potato. Mol Gen Genet 246:745–755
Linsmaier EM, Skoog F (1965) Organic growth factor requirement of tobacco tissue culture. Physiol Plant 18:100–127
Noda T, Takahata Y, Sato T, Kumagai T, Yamakawa O (1998) Starch properties and and cell-wall material contents in sweet potatos as affected by flesh color, cultivation method and year. Jpn App Glycosci 45:1–9
Noda T, Kimura T, Otani M, Ideta O, Shimada T, Saito A, Suda I (2002) Physicochemical properties of amylase-free starch from transgenic sweet potato. Carbohydr Polym 49:253–260
Ogita S, Uefuji H, Morimoto M, Sano H (2004) Application of RNAi to confirm theobromine as the major intermediate for caffeine biosynthesis in coffee plants with potential for construction decaffeinated varieties. Plant Mol Biol 54:931–941
Otani M, Shimada T (1996) Efficient embryogenic callus formation in sweet potato (Ipomoea batatas (L.) Lam.). Breed Sci 46:275–280
Otani M, Shimada T, Kimura T, Saito A (1998) Transgenic plant production from embryogenic callus of sweet potato (Ipomoea batatas (L.) Lam.) using Agrobacterium tumefaciens. Plant Biotechnol 15:11–16
Otani M, Wakita Y, Shimada T (2003) Production of herbicide-resistant sweet potato (Ipomoea batatas (L) Lam.) plants by Agrobacterium tumefaciens-mediated transformation. Breed Sci 53:145–148
Shimada H, Tada Y, Kawasaki T, Fujimura T (1993) Antisense regulation of the rice waxy gene expression using a PCR-amplified fragment of the rice genome reduces the amylose content in grain starch. Theo Appl Genet 86:665–672
Shimada T, Otani M, Hamada T, Kim SH (2006) Increase of amylose content of sweet potato starch by RNA interference of the starch branching enzyme II gene (IbSBEII). Plant Biotechnol 23:85–90
Stoutjesdijk PA, Singh SP, Liu Q, Hurlstone CJ, Waterhouse PA, Green AG (2002) hp-RNA-mediated targeting of the Arabidopsis FAD2 gene gives highly efficient and stable silencing. Plant Physiol 129:1723–1731
Visser RGF, Somhorst I, Kuipers GJ, Ruys NJ, Feenstra WJ, Jacobson E (1991) Inhibition of the expression of the gene for granule-bound starch synthase in potato by antisense constructs. Mol Gen Genet 225:289–296
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Communicated by R. Schmidt.
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Otani, M., Hamada, T., Katayama, K. et al. Inhibition of the gene expression for granule-bound starch synthase I by RNA interference in sweet potato plants. Plant Cell Rep 26, 1801–1807 (2007). https://doi.org/10.1007/s00299-007-0396-6
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DOI: https://doi.org/10.1007/s00299-007-0396-6