Abstract
Soybean (Glycine max (L.) Merr.) is an important cultivated crop, which requires much water during its growth, and drought seriously affects soybean yields. Studies have shown that the expression of small heat shock proteins can enhance drought resistance, cold resistance and salt resistance of plants. In this experiment, soybean GmHsps_p23-like gene was successfully cloned by RT-PCR, the protein encoded by the GmHsps_p23-like gene was subjected to bioinformatics analysis, and the pCAMBIA3301-GmHsps_p23-like overexpression vector and pCBSG015-GmHsps_p23-like gene editing vector were constructed. Agrobacterium-mediated method was used to transform soybeans to obtain positive plants. RT-PCR detection, rehydration experiment and drought resistance physiological and biochemical index detection were performed on the T2 generation positive transgenic soybean plants identified by PCR and Southern hybridization. The results showed that the overexpression vector plant GmHsps_p23-like gene expression increased. After rehydration, the transgenic overexpression plants returned to normal growth, and the damage to the plants was low. After drought stress, the SOD and POD activities and the PRO content of the transgenic overexpression plants increased, while the MDA content decreased. The reverse was true for soybean plants with genetically modified editing vectors. The drought resistance of the overexpressed soybeans under drought stress was higher than that of the control group, and had a stronger drought resistance. It showed that the expression of soybean GmHsps_p23-like gene can improve the drought resistance of soybean. The cloning and functional verification of soybean GmHsps_p23-like gene had not been reported yet. This is the first time that PCR technology has been used to amplify the soybean GmHsps_p23-like gene and construct an expression vector for this gene. This research has laid the foundation for transgenic technology to improve plant drought resistance and cultivate new drought-resistant transgenic soybean varieties.
Highlights
During growth of soybean, the demand for water is high, and drought seriously affects soybean yield.The annual loss of soybean yield and quality in China is immeasurable [1,2]
3.1 Cloning and Identification of Soybean GmHsps_p23-Like Gene The extracted total RNA of soybeans was detected by 2% agarose gel electrophoresis (Fig. 2), and the good quality total RNA was reverse transcribed to generate cDNA
By constructing the Hsp17.4 gene overexpression vector and RNAi interference vector were constructed respectively, and the relevant indexes were measured after genetic transformation of soybean
Summary
The demand for water is high, and drought seriously affects soybean yield. The annual loss of soybean yield and quality in China is immeasurable [1,2]. As a major grain producing area, the cultivation and production of soybean in Northeast China is greatly affected by extreme weather. 2022, vol., no.6 events, such as high temperatures and drought [3]. Improving plant drought resistance through transgenic technology has shown great application and economic value. Cloning of genes related to drought resistance and transferring them into the genome is one of the most effective methods to cultivate drought-tolerant soybean varieties
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