Overexpression of VaBAM3 from Vitis amurensis enhances seedling cold tolerance by promoting soluble sugar accumulation and reactive oxygen scavenging

• Published in: Plant cell reports Vol. 43; no. 6; p. 151
• Main Authors: Liang, Guoping, Wang, Han, Gou, Huimin, Li, Min, Cheng, Yongjuan, Zeng, Baozhen, Mao, Juan, Chen, Baihong
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-06-2024; Springer Nature B.V
• Subjects: Accumulation; Amylases; Arabidopsis; Arabidopsis - genetics; Arabidopsis - metabolism; Arabidopsis - physiology; beta-Amylase - genetics; beta-Amylase - metabolism; Biomedical and Life Sciences; Biotechnology; Callus; Catalase; Cell Biology; Cloning; Cold; Cold Temperature; Cold tolerance; Cold-Shock Response - genetics; Cold-Shock Response - physiology; COR gene; CRISPR; Cultivation; Extreme cold; Freezing; Fruit cultivation; Fruits; Gene expression; Gene Expression Regulation, Plant; Germplasm; Grapes; Life Sciences; Low temperature resistance; Original Article; Oxygen; Peroxidase; Phagosomes; Plant Biochemistry; Plant Proteins - genetics; Plant Proteins - metabolism; Plant Sciences; Plants, Genetically Modified; Reactive oxygen species; Reactive Oxygen Species - metabolism; Scavenging; Seedlings; Seedlings - genetics; Seedlings - physiology; Solanum lycopersicum - genetics; Solanum lycopersicum - metabolism; Solanum lycopersicum - physiology; Starch; Starch - metabolism; Sucrose; Sugar; Sugars - metabolism; Superoxide dismutase; Tomatoes; Vitis - genetics; Vitis - metabolism; Vitis - physiology; Vitis amurensis; Wilting; β-Amylase; Callus; Cold stress; CRISPR/Cas9; Grape; VaBAM3
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-024-03236-5

Key message
The VaBAM3 cloned from Vitis amurensis can enhance the cold tolerance of overexpressed plants, but VaBAM3 knock out by CRISPR/Cas9 system weakened grape callus cold tolerance.
In grape production, extreme cold conditions can seriously threaten plant survival and fruit quality. Regulation of starch content by β-amylase (BAM, EC: 3.2.1.2) contributes to cold tolerance in plants. In this study, we cloned the
VaBAM3
gene from an extremely cold-tolerant grape,
Vitis amurensis
, and overexpressed it in tomato and
Arabidopsis
plants, as well as in grape callus for functional characterization. After exposure to cold stress, leaf wilting in the
VaBAM3
-overexpressing tomato plants was slightly less pronounced than that in wild-type tomato plants, and these plants were characterized by a significant accumulation of autophagosomes. Additionally, the
VaBAM3
-overexpressing
Arabidopsis
plants had a higher freezing tolerance than the wild-type counterparts. Under cold stress conditions, the activities of total amylase, BAM, peroxidase, superoxide dismutase, and catalase in
VaBAM3
-overexpressing plants were significantly higher than those in the corresponding wild-type plants. Furthermore, sucrose, glucose, and fructose contents in these lines were similarly significantly higher, whereas starch contents were reduced in comparison to the levels in the wild-type plants. Furthermore, we detected high
CBF
and
COR
gene expression levels in cold-stressed
VaBAM3
-overexpressing plants. Compared with those in
VaBAM3
-overexpressing grape callus, the aforementioned indicators tended to change in the opposite direction in grape callus with silenced
VaBAM3
. Collectively, our findings indicate that heterologous overexpression of
VaBAM3
enhanced cold tolerance of plants by promoting the accumulation of soluble sugars and scavenging of excessive reactive oxygen species. These findings provide a theoretical basis for the cultivation of cold-resistant grape and support creation of germplasm resources for this purpose.