Inhibition of the gene expression for granule-bound starch synthase I by RNA interference in sweet potato plants |
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Authors: | Motoyasu Otani Tatsuro Hamada Kenji Katayama Kakefumi Kitahara Sun-Hyung Kim Yasuhiro Takahata Toshihiko Suganuma Takiko Shimada |
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Affiliation: | (1) Research Institute of Agricultural Resources, Ishikawa Prefectural University, Nonoichi, Ishikawa 921-8836, Japan;(2) National Agricultural Research Center for Kyushu Okinawa Region, 6644 Yokoichi, Miyakonojo, Miyazaki 885-0091, Japan;(3) Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan;(4) Present address: Department of Environmental Horticulture, University of Seoul, 90 jeonnong-dong, Dongdaemun-gu, Seoul, 130-743, Korea;(5) Present address: Department of Plant Sciences, University of Colombo, P.O. Box 1490, Colombo, 00300, Sri Lanka |
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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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Keywords: | Sweet potato Transgenic plant Amylose free GBSSI |
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