Fen-Xia Han1, Bao-Qing Dun1, Ji Zhang1, Zhi Wang1, Yi Sui1, Li Zhu2, Gui-Ying Li3. 1. National Engineering Laboratory for Crop Molecular Breeding, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. 2. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. zhuli01@caas.cn. 3. National Engineering Laboratory for Crop Molecular Breeding, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. liguiying@caas.cn.
Abstract
MAIN CONCLUSION: The sorghum soluble acid invertase gene SbSAI-2 was cloned and the function verified in Pichia pastoris and rice, showing the SbSAI-2 affects composition and content of sugar in stem juice. Sugar metabolism is one of the most important metabolic processes in plants, in which soluble acid invertase plays a key role. However, the structure and function of the soluble acid transferase gene in sorghum are still fully unclear. In this study, SbSAI-2 was cloned from the sorghum variety BTx623, and two transcripts were found through sequence analysis, with only one transcript translated into an active protein. There is 72% homology between SbSAI-2 and OsVIN2. The construction of Osvin2 mutant lines and SbSAI-2-1 overexpression lines in Oryza sativa L. japonica. cv. Nipponbare were produced to clarify the invertase functionality. While the invertase activity in the stem of the Osvin2 mutant line was reduced, with no significant difference (P > 0.05), and the contents of fructose and glucose in stem tissue did not change significantly (P > 0.05), and the content of sucrose increased by 38.89% (P < 0.01). In SbSAI-2-1 overexpression lines, the invertase activity in stem was increased by more than 20 times (P < 0.01). The contents of glucose and fructose in stem tissues were increased by two and three times, respectively (P < 0.01), while the content of sucrose was significantly decreased, which was below the detection limit (P < 0.01). This study indicated that SbSAI-2 is a key enzyme related to sucrose metabolism and affects the composition and content of sugar in stems. The result provided further the gene function verification and laid a foundation for the development of molecular markers.
MAIN CONCLUSION: The sorghum soluble acid invertase gene SbSAI-2 was cloned and the function verified in Pichia pastoris and rice, showing the SbSAI-2 affects composition and content of sugar in stem juice. Sugar metabolism is one of the most important metabolic processes in plants, in which soluble acid invertase plays a key role. However, the structure and function of the soluble acid transferase gene in sorghum are still fully unclear. In this study, SbSAI-2 was cloned from the sorghum variety BTx623, and two transcripts were found through sequence analysis, with only one transcript translated into an active protein. There is 72% homology between SbSAI-2 and OsVIN2. The construction of Osvin2 mutant lines and SbSAI-2-1 overexpression lines in Oryza sativa L. japonica. cv. Nipponbare were produced to clarify the invertase functionality. While the invertase activity in the stem of the Osvin2 mutant line was reduced, with no significant difference (P > 0.05), and the contents of fructose and glucose in stem tissue did not change significantly (P > 0.05), and the content of sucrose increased by 38.89% (P < 0.01). In SbSAI-2-1 overexpression lines, the invertase activity in stem was increased by more than 20 times (P < 0.01). The contents of glucose and fructose in stem tissues were increased by two and three times, respectively (P < 0.01), while the content of sucrose was significantly decreased, which was below the detection limit (P < 0.01). This study indicated that SbSAI-2 is a key enzyme related to sucrose metabolism and affects the composition and content of sugar in stems. The result provided further the gene function verification and laid a foundation for the development of molecular markers.
Authors: Xuemei Ji; Wim Van den Ende; Lindsey Schroeven; Stefan Clerens; Koen Geuten; Shihua Cheng; John Bennett Journal: New Phytol Date: 2007 Impact factor: 10.151
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