The sense and antisense fragments of the soluble starch synthase （SSII1） gene and the intron fragment of somatic embryogenesis receptor-like kinase （SERK1） gene were cloned from potato using PCR techniques. The RN...The sense and antisense fragments of the soluble starch synthase （SSII1） 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 SSlll gene was integrated into potato genome. The RT-PCR analysis showed that the expression of SSlll 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.展开更多
基金supported by the National High-Technology R&D Program of China(2006AA100107)the National Natural Science Foundation of China(30871573, 31160298)
文摘The sense and antisense fragments of the soluble starch synthase （SSII1） 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 SSlll gene was integrated into potato genome. The RT-PCR analysis showed that the expression of SSlll 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.
