Overexpression of 2-Cys Peroxiredoxin alleviates the NaHCO3 stress-induced photoinhibition and reactive oxygen species damage of tobacco.

Plant 2-cysteine peroxiredoxin (2-Cys Prx) is a mercaptan peroxidase localized in chloroplasts and has unique catalytic properties. To explore the salt stress tolerance mechanisms of 2-Cys Prx in plants, we analyzed the effects of overexpressing the 2-CysPrx gene on the physiological and biochemical metabolic processes of tobacco under NaHCO3 stress through joint physiological and transcriptomic analysis. These parameters included growth phenotype, chlorophyll, photosynthesis, and antioxidant system. After NaHCO3 stress treatment, a total of 5360 differentially expressed genes (DEGs) were identified in 2-Cysprx overexpressed (OE) plants, and the number of DEGs was significantly lower than 14558 in wild-type (WT) plants. KEGG enrichment analysis showed that DEGs were mainly enriched in photosynthetic pathways, photosynthetic antenna proteins, and porphyrin and chlorophyll metabolism. Overexpressing 2-CysPrx significantly reduced the growth inhibition of tobacco induced by NaHCO3 stress, alleviating the down-regulation of the DEGs related to chlorophyll synthesis, photosynthetic electron transport and the Calvin cycle and the up-regulation of those related to chlorophyll degradation. In addition, it also interacted with other redox systems such as thioredoxins (Trxs) and the NADPH-dependent Trx reductase C (NTRC), and mediated the positive regulation of the activities of antioxidant enzymes such as peroxidase (POD) and catalase (CAT) and the expression of related genes, thereby reducing the accumulation of superoxide anion (O2·-), hydrogen peroxide (H2O2) and malondialdehyde (MDA). In conclusion, 2-CysPrx overexpression could alleviate the NaHCO3 stress-induced photoinhibition and oxidative damage by regulating chlorophyll metabolism, promoting photosynthesis and participating in the regulation of antioxidant enzymes, and thus improve the ability of plants to resist salt stress damage.

Heterologous expression of Zygophyllum xanthoxylon zinc finger protein gene (ZxZF) enhances the tolerance of poplar photosynthetic function to drought stress.

The ZxZF transcription factor (TF) of Zygophyllum xanthoxylon (Bunge) Maxim, an extremely drought-resistant woody plant, is a C2H2 zinc finger protein. Studies have shown that C2H2 zinc finger proteins play important roles in activating stress-related genes and enhancing plant resistance. However, their function in regulating plant photosynthesis under drought stress is not well understood. Since poplar is an important greening and afforestation tree species, it is particularly important to cultivate excellent drought-tolerant varieties. The ZxZF transcription factor (TF) was heterogeneously expressed in Euroamerican poplar (Populus × euroameracana cl.'Bofengl') by genetic transformation. Based on the mechanism and potential function of poplar photosynthesis regulated by ZxZF under drought stress, transcriptomic and physiological determinations were used to reveal the important role of this gene in improving the drought resistance of poplar. The results showed that the overexpression of ZxZF TF in transgenic poplars could improve the inhibition of Calvin cycle by regulating stomatal opening and increasing the concentration of intercellular CO2. The chlorophyll content, photosynthetic performance index, and photochemical efficiency of transgenic lines under drought stress were significantly higher than those of the wild type (WT). The overexpression of ZxZF TFs could alleviate the degree of photoinhibition of photosystems II and I under drought stress and maintain the efficiency of light energy capture and the photosynthetic electron transport chain. The transcriptomic data also showed that differentially expressed genes between the transgenic poplar and WT under drought stress were primarily enriched in metabolic pathways related to photosynthesis, such as photosynthesis, photosynthesis-antenna protein, porphyrin and chlorophyll metabolism, and photosynthetic carbon fixation, and the downregulation of genes related to chlorophyll synthesis, photosynthetic electron transport and Calvin cycle were alleviated. In addition, the overexpression of ZxZF TF can alleviate the inhibition of NADH dehydrogenase-like (NDH) cyclic electron flow of the poplar NDH pathway under drought stress, which plays an important role in reducing the excess pressure of electrons on the photosynthetic electron transport chain and maintaining the normal photosynthetic electron transport. In summary, the overexpression of ZxZF TFs can effectively alleviate the inhibition of drought on the assimilation of carbon in poplar and have a positive impact on light energy capture, the orderly transport of photosynthetic electron transport chain and the integrity of the photosystem, which is highly significant to acheivean in-depth understanding of the function of ZxZF TFs. This also provides an important basis for the breeding of new transgenic poplar varieties.

Transcriptome and functional analysis revealed the intervention of brassinosteroid in regulation of cold induced early flowering in tobacco.

Cold environmental conditions may often lead to the early flowering of plants, and the mechanism by cold-induced flowering remains poorly understood. Microscopy analysis in this study demonstrated that cold conditioning led to early flower bud differentiation in two tobacco strains and an Agilent Tobacco Gene Expression microarray was adapted for transcriptomic analysis on the stem tips of cold treated tobacco to gain insight into the molecular process underlying flowering in tobacco. The transcriptomic analysis showed that cold treatment of two flue-cured tobacco varieties (Xingyan 1 and YunYan 85) yielded 4176 and 5773 genes that were differentially expressed, respectively, with 2623 being commonly detected. Functional distribution revealed that the differentially expressed genes (DEGs) were mainly enriched in protein metabolism, RNA, stress, transport, and secondary metabolism. Genes involved in secondary metabolism, cell wall, and redox were nearly all up-regulated in response to the cold conditioning. Further analysis demonstrated that the central genes related to brassinosteroid biosynthetic pathway, circadian system, and flowering pathway were significantly enhanced in the cold treated tobacco. Phytochemical measurement and qRT-PCR revealed an increased accumulation of brassinolide and a decreased expression of the flowering locus c gene. Furthermore, we found that overexpression of NtBRI1 could induce early flowering in tobacco under normal condition. And low-temperature-induced early flowering in NtBRI1 overexpression plants were similar to that of normal condition. Consistently, low-temperature-induced early flowering is partially suppressed in NtBRI1 mutant. Together, the results suggest that cold could induce early flowering of tobacco by activating brassinosteroid signaling.

Trx CDSP32-overexpressing tobacco plants improves cadmium tolerance by modulating antioxidant mechanism.

The effects of overexpression of the thioredoxin-like protein CDSP32 (Trx CDSP32) on reactive oxygen species (ROS) metabolism in tobacco leaves exposed to cadmium (Cd) were studied by combining physiological measures and proteomics technology. Thus, the number of differentially expressed proteins (DEPs) in plants overexpressing the Trx CDSP32 gene in tobacco (OE) was observed to be evidently lower than that in wild-type (WT) tobacco under Cd exposure, especially the number of down-regulated DEPs. Cd exposure induced disordered ROS metabolism in tobacco leaves. Although Cd exposure inhibited the activities of superoxide dismutase (SOD), catalase (CAT), and l-ascorbate peroxidase (APX) and the expression of proteins related to the thioredoxin-peroxiredoxin (Trx-Prx) pathway, the increase in the activities of peroxidase (POD), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione peroxidase (GPX), and glutathione S-transferase (GST) and their protein expression levels played an important role in the physiological response to Cd exposure. Notably, Trx CDSP32 was observed to alleviate the decrease in the expression and activities of SOD and CAT caused by Cd exposure and enhance the function of POD. Trx CDSP32 was observed to increase the H2O2 scavenging capacity of the ascorbic acid-glutathione (AsA-GSH) cycle and Trx-Prx pathway under Cd exposure, and it can especially regulate 2-Cys peroxiredoxin (2-Cys Prx) protein expression and thioredoxin peroxidase (TPX) activity. Thus, overexpression of the Trx CDSP32 gene can alleviate the oxidative damage that occurs in tobacco leaves under Cd exposure by modulating antioxidant defense systems.