Cloning of Sugar Transporter MeSWEET2b in Cassava and Its Function Analysis Under Low Temperature Stress

doi:10.13560/j.cnki.biotech.bull.1985.2025-0857

Abstract

Abstract:

Objective To study the function of the MeSWEET2b gene in cassava (Manihot esculenta Crantz) under low-temperature stress, and to provide theoretical basis for exploring the molecular mechanism of cassava tolerance to cold and breeding new varieties with low-temperature tolerance. Method Bioinformatics method was used to analyze the sequence characteristics, phylogenetic relationships of the MeSWEET2b gene. Transient expression of the MeSWEET2b protein in tobacco was to determine its subcellular localization. RT-qPCR was adapted to analyze the expressions of the MeSWEET2b gene at different growth stages, under darkness treatments of different sugar and low-temperature stress. Virus induced gene silencing (VIGS) technology was applied to study the function of this gene under low-temperature stress. Result The MeSWEET2b protein had 7 transmembrane domains, and phylogenetic tree analysis indicated that it belonged to the SWEETs Clade I subfamily and had the closest homology with Arabidopsis AtSWEET2. The subcellular localization results showed that MeSWEET2b was located on the cell plasma membrane. The RT-qPCR analysis showed that MeSWEET2b had the highest expression in the leaves during the expansion period (7 months). For the water culture seedlings of cassava treated with different sugars, the expression trends of MeSWEET2b in glucose and fructose were consistent. As the temperature of cold stress decreased and the time extended, the expression of MeSWEET2b in cassava showed a significantly increased. Using VIGS technology to silence MeSWEET2b, under 4 ℃ low-temperature stress, the wilting of the leaves in the MeSWEET2b silenced plants were significantly more severe than that of the control, and the contents of glucose and fructose significantly reduced. Conclusion The silencing of MeSWEET2b gene led to the intolerance of cassava to low temperatures. It is speculated that this gene plays an important role in the cold tolerance of cassava.

Key words: cassava, sugar transporter, MeSWEET2b, low temperature stress, VIGS

XUE Jing-jing, AN Fei-fei, LUO Xiu-qin, CAI Jie. Cloning of Sugar Transporter MeSWEET2b in Cassava and Its Function Analysis Under Low Temperature Stress[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2025-0857.

Figures/Tables 10

Fig. 1 CDS sequences and amino acid sequences of MeSWEET2b* indicates stop codon, underline indicates conserved domains MtN3_slv super family, bold font indicates transmembrane sequence

Fig. 2 Phylogenetic analysis and acid sequence alignment of MeSWEET2b

Fig. 3 Subcellular localization of MeSWEET2b

Fig. 4 Expression analysis of MeSWEET2b at different developmental stages

Fig. 5 Expression analysis of MeSWEET2b in dark treatment with different sugar solutions

Fig. 6 Expression analysis of MeSWEET2b under low temperature stress

Fig. 7 Expression level analysis of cassava MeSWEET2b silenced plants

Fig. 8 Phenotypic characteristics of cassava MeSWEET2b silenced plants under low-temperature stress

Fig. 9 Sugar content of cassava MeSWEET2b silenced plants under low-temperature stress

Fig. 10 Analysis of physiological indicators in cassava MeSWEET2b silenced plants under low-temperature stress

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