AUTHOR=Zhang Xiaomeng , Jiang Jingwen , Ma Zewang , Yang Yupeng , Meng Lingdong , Xie Fuchun , Cui Guowen , Yin Xiujie 

TITLE=Cloning of TaeRF1 gene from Caucasian clover and its functional analysis responding to low-temperature stress

JOURNAL=Frontiers in Plant Science

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2022.968965

DOI=10.3389/fpls.2022.968965

ISSN=1664-462X

ABSTRACT=<p>Low temperature (LT) is an important threat to the normal growth of plants. In this study, based on the full-length transcriptome sequencing results, the cold resistance genes were cloned from Caucasian clover with strong cold resistance. We cloned the CDS of <italic>TaeRF1</italic>, which is 1311 bp in length and encodes 436 amino acids. The molecular weight of the protein is 48.97 kDa, which had no transmembrane structure, and its isoelectric point (pI) was 5.42. We predicted the structure of <italic>TaeRF1</italic> and found 29 phosphorylation sites. Subcellular localization showed that <italic>TaeRF1</italic> was localized and expressed in cell membrane and chloroplasts. The <italic>TaeRF1</italic> gene was induced by stress due to cold, salt, alkali and drought and its expression level was higher in roots and it was more sensitive to LT. Analysis of transgenic A. thaliana plants before and after LT treatment showed that the <italic>TaeRF1</italic> gene enhanced the removal of excess H<sub>2</sub>O<sub>2</sub>, and increased the activity of antioxidant enzymes, thus improving the plant’s ability to resist stress. Additionally, the OE lines showed increased cold tolerance by upregulating the transcription level of cold-responsive genes (<italic>CBF1</italic>, <italic>CBF2</italic>, <italic>COR15B</italic>, <italic>COR47</italic>, <italic>ICE1</italic>, and <italic>RD29A</italic>). This study demonstrates that <italic>TaeRF1</italic> is actively involved in the responses of plants to LT stress. We also provide a theoretical basis for breeding and a potential mechanism underlying the responses of Caucasian clover to abiotic stress.</p>