This study evaluated AtbZIP69 overexpression in wheat material and verified that it could improve low-nitrogen and drought tolerance in wheat. Our results showed that the expression of AtbZIP69 was upregulated by 20-fold under low-nitrogen stress and several-fold under drought stress. This suggests that AtbZIP69 responds to drought and LN stress in wheat (Fig. 2). This was consistent with previous findings in Arabidopsis. This suggests that the function of AtbZIP69 is conserved across species. Numerous studies have confirmed that bZIP TFs play important roles in the responses of many species to drought and other abiotic stresses (Liu et al., 2014; Yang et al., 2020). Transgenic wheat performed significantly better than the WT under drought stress, regardless of biomass, survival rate, or plant height stress index (Fig. 4). Although several studies have found that bZIP TF is associated with the nitrate transport pathway, there are relatively few studies on the role of bZIP TF in improving low-nitrogen tolerance. Please add some scientific discussion to make the content more explanatory to the scientific questions The findings of this study provide valuable insights into the potential use of AtbZIP69 overexpression in improving low-nitrogen and drought tolerance in wheat. The upregulation of AtbZIP69 gene expression in response to low-nitrogen and drought stress suggests its involvement in the stress response mechanisms of wheat. The observed increase in AtbZIP69 expression by 20-fold under low-nitrogen stress and several-fold under drought stress indicates that this gene is highly responsive to these abiotic stresses. This responsiveness is consistent with previous findings in Arabidopsis, suggesting that the function of AtbZIP69 is conserved across species. This conservation of function highlights the significance of AtbZIP69 in stress signaling pathways. The improved performance of transgenic wheat overexpressing AtbZIP69 under drought stress, as demonstrated by various parameters such as biomass, survival rate, and plant height stress index, further supports the potential of AtbZIP69 in enhancing drought tolerance in wheat. These results suggest that AtbZIP69 overexpression can enhance the adaptive capacity of wheat plants under drought conditions. Furthermore, while previous studies have explored the role of bZIP TFs in drought response in various species, there is relatively limited research on their involvement in improving low-nitrogen tolerance. This study provides evidence that AtbZIP69 overexpression can improve low-nitrogen tolerance in wheat, which adds to our understanding of the complex regulatory mechanisms underlying nitrogen stress responses. Overall, these findings highlight the potential of AtbZIP69 as a key player in enhancing both low-nitrogen and drought tolerance in wheat. Further investigation into the specific mechanisms through which AtbZIP69 functions in stress responses, as well as its interaction with other genes and pathways, could provide additional insights into its potential applications in crop improvement strategies.


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