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.
Abstract
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.
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Data availability
The data that support the findings of this study are available from the corresponding authors upon reasonable request.
Abbreviations
- AS:
-
Alternative splicing
- GH32:
-
Glycoside hydrolase family 32
- SAI:
-
Soluble acid invertase
- sgRNA:
-
Single guide RNA
- VIN:
-
Vacuolar invertase
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This research is supported in part by National Key Research & Development Program (2018YFD10007002, 2019YFD1001700), Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences, International Atomic Energy Agency/CR Project (D23031).
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Supplementary file 2: Suppl. Fig.S2 Comparison of sugar content in different rice lines. The peak time of fructose is 3.5–4.0 min, the peak time of glucose is 4.0–4.5 min, and the peak time of sucrose is 5.0–5.5 min. Osvin2 is a mutant line at OsVIN2 locus from the wild type (WT); 1390-SbSAI-2-1 is the SbSAI-2-1 overexpressing line. LSU is the abbreviation of Laser Scanning Unit. (DOCX 178 KB)
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Wuyuntanmanda, Han, FX., Dun, BQ. et al. Cloning and functional analysis of soluble acid invertase 2 gene (SbSAI-2) in sorghum. Planta 255, 13 (2022). https://doi.org/10.1007/s00425-021-03772-4
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DOI: https://doi.org/10.1007/s00425-021-03772-4