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| Effects of RNAi Silencing of SSIII Gene on Phosphorus Content and Characteristics of Starch in Potato Tubers |
| DU Hong-hui, YANG Tao, MA Cong-yu, FENG Dan, ZHANG Ning, SI Huai-jun, WANG Di |
1.Gansu Provincial Key Laboratory of Aridland Crop Science, Gansu Key Laboratory of Crop Genetic and Germplasm Enhancement,Lanzhou 730070, P.R.China
2.College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, P.R.China
3.Biological Research Institute, Gansu Academy of Sciences, Lanzhou 730000, P.R.China |
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摘要 The sense and antisense fragments of the soluble starch synthase (SSIII) gene and the intron fragment of somatic embryogenesis receptor-like kinase (SERK1) gene were cloned from potato using PCR techniques. The RNAi plant expression vectors pBI-SSIII-RNAi and pBIC-SSIII-RNAi were constructed which containing fusion fragment of “sense fragment-intron-antisense fragment” driven by the constitutive expression promoter CaMV 35S and the tuber-specific expression promoter CIPP, respectively. The putative transgenic plants of potato cultivars Kexin-1 and Kexin-4 were obtained using Agrobacterium-mediated transformation method. PCR assay showed that the interference fragment of SSIII gene was integrated into potato genome. The RT-PCR analysis showed that the expression of SSIII gene was repressed apparently on the transcription level. Starch granules of the transgenic potato plants were different in morphology and became cracked in starch granule centre compared with the non-transgenic control plants. The amylose content of starch was increased by 2.68-29.05%, amylopectin to amylose ratio of starch had declined significantly, and the phosphorus content of the starch of the transgenic plants was reduced 9.94-58.36% compared with control plants. The results could provide certain foundation for improvement of potato starch quality.
Abstract The sense and antisense fragments of the soluble starch synthase (SSIII) gene and the intron fragment of somatic embryogenesis receptor-like kinase (SERK1) gene were cloned from potato using PCR techniques. The RNAi plant expression vectors pBI-SSIII-RNAi and pBIC-SSIII-RNAi were constructed which containing fusion fragment of “sense fragment-intron-antisense fragment” driven by the constitutive expression promoter CaMV 35S and the tuber-specific expression promoter CIPP, respectively. The putative transgenic plants of potato cultivars Kexin-1 and Kexin-4 were obtained using Agrobacterium-mediated transformation method. PCR assay showed that the interference fragment of SSIII gene was integrated into potato genome. The RT-PCR analysis showed that the expression of SSIII gene was repressed apparently on the transcription level. Starch granules of the transgenic potato plants were different in morphology and became cracked in starch granule centre compared with the non-transgenic control plants. The amylose content of starch was increased by 2.68-29.05%, amylopectin to amylose ratio of starch had declined significantly, and the phosphorus content of the starch of the transgenic plants was reduced 9.94-58.36% compared with control plants. The results could provide certain foundation for improvement of potato starch quality.
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Received: 20 July 2011
Accepted:
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| Fund: This work was supported by the National High-Technology R&D Program of China (2006AA100107) and the National Natural Science Foundation of China (30871573, 31160298). |
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Corresponding Authors:
Correspondence SI Huai-jun, Tel: +86-931-7631077, E-mail: hjsi@gsau.edu.cn; WANG Di, Tel: +86-931-7632826, E-mail: wangd@gsau.edu.cn
E-mail: hjsi@gsau.edu.cn
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| About author: DU Hong-hui, Mobile: 18009325320, E-mail: duhonghui2008@163.com; |
Cite this article:
DU Hong-hui, YANG Tao, MA Cong-yu, FENG Dan, ZHANG Ning, SI Huai-jun, WANG Di.
2012.
Effects of RNAi Silencing of SSIII Gene on Phosphorus Content and Characteristics of Starch in Potato Tubers. Journal of Integrative Agriculture, 12(12): 1985-1992.
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