Overexpression of sugarcane SoTUA gene enhances cold tolerance in transgenic sugarcane

Abstract

Alpha (α)-tubulin (TUA) is a primary unit of cytoskeleton microtubules. In recent years, more frequent frosts have caused considerable yield loss in sugarcane (Saccharum spp. hybrid) production. Temperature is one of the most influential abiotic factors affecting the microtubule polymerization state in plants. In this research, we developed overexpressing SoTUA transgenic sugarcane lines from the cold-susceptible cultivar ROC22. The transgene was confirmed by polymerase chain reaction (PCR) and quantificational real-time polymerase chain reaction (qRT-PCR) analysis, while the relative expression of SoTUA protein was verified by Western blot. Compared with the nontransformed control plants (wild type [WT]), the transgenic lines showed higher relative protein expression than the WT at 25 ˚C, which indicates the overexpression of SoTUA protein. The contents of soluble protein and sugar and the activity of peroxidase (POD) were higher while the content of maleicdialdehyde was lower in the transgenic plants than the WT plants under the chilling treatment. Meanwhile, the expression levels of cold-defense related genes (P5CS, WRKY, Cu/Zn-SOD, and POD1) were higher in the transgenic lines than in the WT. These results suggest that the SoTUA protein plays an important role in protecting plants during chilling stress. This positive effect is not affected by transgenic plant generations.

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“…The relative level of gene expression was determined using the 2 ÀΔΔCt formula. GADPH was used as the internal reference (Chen et al, 2021). The primer sequences are listed in Table S1.…”

Genetic Engineering for Enhancing Sugarcane Tolerance to Biotic and Abiotic Stresses

“…Compared with nontransgenic parent plants exposed to low temperatures, transgenic sugarcane exhibited increased chlorophyll, reduced MDA content, and decreased electrolyte leakage [109]. Transgenic plants overexpressing the tubulin gene (TUA), cloned from cold-tolerant sugarcane varieties, enhanced the cold tolerance of cold-susceptible sugarcane varieties, exhibiting increased soluble protein and sugar content, enhanced peroxidase activity, and reduced MDA content compared to non-transgenic plants [110]. Further investigation into the effects of cold stress could have substantial implications for the sugarcane industry, as cold stress negatively affects sugarcane crop yields and quality.…”

Genetic engineering: an efficient approach to mitigating biotic and abiotic stresses in sugarcane cultivation

“…The transgenic sugarcane overexpressing a bacterial isopentenyl-transferase gene ( ipt ) and using a cold-inducible promoter, At COR15 a from A. thaliana , showed increased leaf chlorophyll content, reduced MDA accumulation, and electrolyte leakage concerning the non-transformed parental plants submitted to low temperatures. 61 Chen et al 62 (2021) reported the transgenic plants overexpressing α-tubulin gene ( TUA ), which was cloned from a cold resistant sugarcane variety, improved the cold resistance of a cold susceptible sugarcane variety, showing higher contents of total soluble proteins and sugars, increased peroxidase activity, and lower MDA accumulation than the non-transformed plants under excessive temperature conditions.…”