ThCOL2 Improves the Salt Stress Tolerance of Tamarix hispida

Abstract

The CONSTANS-LIKE (COL) transcription factor has been reported to play important roles in regulating plant flowering and the response to abiotic stress. To clone and screen <i>COL</i> genes with excellent salt tolerance from the woody halophyte <i>Tamarix hispida</i>, 8 <i>ThCOL</i> genes were identified in this study. The expression patterns of these genes under different abiotic stresses (high salt, osmotic, and heavy metal) and abscisic acid (ABA) treatment were detected using quantitative real-time PCR (qRT-PCR). The expression levels of 8 <i>ThCOL</i> genes changed significantly after exposure to one or more stresses, indicating that these genes were all stress-responsive genes and may be involved in the stress resistance response of <i>T. hispida</i>. In particular, the expression level of <i>ThCOL2</i> changed significantly at most time points in the roots and leaves of <i>T. hispida</i> under salt stress and after ABA treatments, which may play an important role in the response process of salt stress through a mechanism dependent on the ABA pathway. The recombinant vectors pROKII-<i>ThCOL2</i> and pFGC5941-<i>ThCOL2</i> were constructed for the transient transformation of <i>T. hispida</i>, and the transient infection of <i>T. hispida</i> with the pROKII empty vector was used as the control to further verify whether the <i>ThCOL2</i> gene was involved in the regulation of the salt tolerance response of <i>T. hispida</i>. Overexpression of the <i>ThCOL2</i> gene in plants under 150 mM NaCl stress increased the ability of transgenic <i>T. hispida</i> cells to remove reactive oxygen species (ROS) by regulating the activity of protective enzymes and promoting a decrease in the accumulation of O^2- and H₂O₂, thereby reducing cell damage or cell death and enhancing salt tolerance. The <i>ThCOL2</i> gene may be a candidate gene associated with excellent salt tolerance. Furthermore, the expression levels of some genes related to the ABA pathway were analyzed using qRT-PCR. The results showed that the expressions of <i>ThNCED1</i> and <i>ThNCED4</i> were significantly higher, and the expressions of <i>ThNCED3</i>, <i>ThZEP</i>, and <i>ThAAO</i>3 were not significantly altered in OE compared with CON under normal conditions. But after 24 h of salt stress, the expressions of all five studied genes all were lower than the normal condition. In the future, the downstream genes directly regulated by the <i>ThCOL2</i> transcription factor will be searched and identified to analyze the salt tolerance regulatory network of <i>ThCOL2</i>.