Characteristics of the chloroplast genome and genetic divergence of Tamarix hispida Willd. 1816 (Tamaricaceae).

Tamarix hispida Willd. 1816, a crucial native plant species in the arid desert region of northwestern China, plays a significant role in maintaining ecological stability. It is instrumental in addressing soil salinity-alkalinity and heavy metal pollution. This research aims to analyze the phylogenetic divergence pattern and evolutionary history of T. hispida by comparing chloroplast genome structures across different populations. Despite the minimal differences in chloroplast genome structure due to conserved genes and junction regions, sequencing was conducted using the Illumina NovaSeq platform to verify the historical evolutionary processes between different populations, followed by assembly and annotation. The results revealed that the T. hispida chloroplast genome is approximately 156,164-156,186 bp in length, with a quadripartite structure and 131 annotated genes. Phylogenetic analysis indicated two lineages within T. hispida, with a divergence time of 3.15 Ma. These findings emphasize the low genetic diversity in T. hispida and offer valuable insights into its evolutionary past. To effectively protect and manage this species, increased scientific research and monitoring of its genetic diversity are necessary. This study underscores the importance of comprehending the genetic mechanisms behind species divergence to develop informed conservation strategies.

The transcription factor ThDOF8 binds to a novel cis-element and mediates molecular responses to salt stress in Tamarix hispida.

Salt stress is a common abiotic factor that restricts plant growth and development. As a halophyte, Tamarix hispida is a good model plant for exploring salt-tolerance genes and regulatory mechanisms. DNA-binding with one finger (DOF) is an important transcription factor (TF) that influences and controls various signaling substances involved in diverse biological processes related to plant growth and development, but the regulatory mechanisms of DOF TFs in response to salt stress are largely unknown in T. hispida. In the present study, a newly identified Dof gene, ThDOF8, was cloned from T. hispida, and its expression was found to be induced by salt stress. Transient overexpression of ThDOF8 enhanced T. hispida salt tolerance by enhancing proline levels, and increasing the activities of the antioxidant enzymes superoxide dismutase (SOD) and peroxidase (POD). These results were also verified in stably transformed Arabidopsis. Results from TF-centered yeast one-hybrid (Y1H) assays and EMSAs showed that ThDOF8 binds to a newly identified cis-element (TGCG). Expression profiling by gene chip analysis identified four potential direct targets of ThDOF8, namely the cysteine-rich receptor-like kinases genes, CRK10 and CRK26, and two glutamate decarboxylase genes, GAD41, and GAD42, and these were further verified by ChIP-quantitative-PCR, EMSAs, Y1H assays, and ß-glucuronidase enzyme activity assays. ThDOF8 can bind to the TGCG element in the promoter regions of its target genes, and transient overexpression of ThCRK10 also enhanced T. hispida salt tolerance. On the basis of our results, we propose a new regulatory mechanism model, in which ThDOF8 binds to the TGCG cis-element in the promoter of the target gene CRK10 to regulate its expression and improve salt tolerance in T. hispida. This study provides a basis for furthering our understanding the role of DOF TFs and identifying other downstream candidate genes that have the potential for improving plant salt tolerance via molecular breeding.

Comparative transcriptomic and metabolomic analyses provide insights into the responses to NaCl and Cd stress in Tamarix hispida.

Salinity and heavy metal pollution seriously affect plant growth. Tamarix hispida (T. hispida) has the potential to remediate soil saline-alkali and heavy metal pollution. In this study, the response mechanisms of T. hispida under NaCl, CdCl2 (Cd) and combined CdCl2 and NaCl (Cd-NaCl) stresses were explored. Overall, the antioxidant system showed changes under the three stresses. The addition of NaCl inhibited the absorption of Cd2+. However, there were obvious differences in the transcripts and metabolites identified among the three stress responses. Interestingly, the number of DEGs was greatest under NaCl stress (929), but the number of differentially expressed metabolites (DEMs) was lowest (48), with 143 and 187 DEMs identified under Cd and Cd-NaCl stress, respectively. It is worth noting that both DEGs and DEMs were enriched in the linoleic acid metabolism pathway under Cd stress. In particular, the content of lipids changed significantly under Cd and Cd-NaCl stress, suggesting that maintaining normal lipid synthesis and metabolism may be an important way to improve the Cd tolerance of T. hispida. Flavonoids may also play an important role in the response to NaCl and Cd stress. These results provide a theoretical basis for cultivating plants with improved salt and cadmium repair abilities.

Overexpression of ThSCL32 confers salt stress tolerance by enhancing ThPHD3 gene expression in Tamarix hispida.

GRAS transcription factors belong to the plant-specific protein family. They are not only involved in plant growth and development but also in plant responses to a variety of abiotic stresses. However, to date, the SCL32(SCARECROW-like 32) gene conferring the desired resistance to salt stresses has not been reported in plants. Here, ThSCL32, a homologous gene of ArabidopsisthalianaAtSCL32, was identified. ThSCL32 was highly induced by salt stress in Tamarix hispida. ThSCL32 overexpression in T. hispida gave rise to improved salt tolerance. ThSCL32-silenced T. hispida plants were more sensitive to salt stress. RNA-seq analysis of transient transgenic T. hispida overexpressing ThSCL32 revealed significantly enhanced ThPHD3 (prolyl-4-hydroxylase domain 3 protein) gene expression. ChIP-PCR further verified that ThSCL32 probably binds to the novel cis-element SBS (ACGTTG) in the promoter of ThPHD3 to activate its expression. In brief, our results suggest that the ThSCL32 transcription factor is involved in salt tolerance in T. hispida by enhancing ThPHD3 expression.

Proteome Dynamics Analysis Reveals the Potential Mechanisms of Salinity and Drought Response during Seed Germination and Seedling Growth in Tamarix hispida.

Understanding the molecular mechanisms of seed germination and seedling growth is vital for mining functional genes for the improvement of plant drought in a desert. Tamarix hispida is extremely resistant to drought and soil salinity perennial shrubs or trees. This study was the first to investigate the protein abundance profile of the transition process during the processes of T. hispida seed germination and seedling growth using label-free proteomics approaches. Our data suggested that asynchronous regulation of transcriptomics and proteomics occurs upon short-term seed germination and seedling growth of T. hispida. Enrichment analysis revealed that the main differentially abundant proteins had significant enrichment in stimulus response, biosynthesis, and metabolism. Two delta-1-pyrroline-5-carboxylate synthetases (P5CS), one Ycf3-interacting protein (Y3IP), one low-temperature-induced 65 kDa protein-like molecule, and four peroxidases (PRX) were involved in both water deprivation and hyperosmotic salinity responses. Through a comparative analysis of transcriptomics and proteomics, we found that proteomics may be better at studying short-term developmental processes. Our results support the existence of several mechanisms that enhance tolerance to salinity and drought stress during seedling growth in T. hispida.

The R2R3-MYB transcription factor ThRAX2 recognized a new element MYB-T (CTTCCA) to enhance cadmium tolerance in Tamarix hispida.

R2R3-MYB transcription factors play an important role in plant development and response to various environmental stresses. In this study, a new R2R3-MYB gene, named ThRAX2, was isolated from T. hispida. ThRAX2 has an open reading frame (ORF) of 1191 bp and encodes a protein of 396 amino acids. ThRAX2 was localized in the nucleus. The overexpression of ThRAX2 in Arabidopsis and T. hispida significantly increased Cadmium (Cd) tolerance. Moreover, the accumulation of cadmium in roots and leaves was significantly reduced. The TF-centred Y1H and Y1H results showed that ThRAX2 was able to specifically bind a new cis-element (MYB-T, CTTCCA). The promoters of some Cd-responsive genes, such as ThSOS1, ThCKX3, ThCAX3A, ThMYB78, ThMIP2, ThTPS4, and ThSOD2, all contained 1-3 MYB-T sequences. Furthermore, chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR) and ChIPquantitative (q)PCR showed that the ThRAX2 gene can bind to ThSOS1, ThCKX3, ThCAX3A and ThMYB78 promoter fragments, including the MYB-T motif. Meanwhile, the qRTPCR results also showed that the expression trends of ThSOS1, ThCKX3, ThCAX3A and ThMYB78 were similar to that of ThRAX2. This finding suggests that Cd tolerance of the ThRAX2 gene may regulate the expression of some downstream genes through specific recognition of the MYB-T motif and participate in regulating intracellular ion homeostasis, transport, and protein activity or enhance antioxidant enzyme activity. This study found a novel cis-acting element that binds ThRAX2 to regulate Cd tolerance, which lays the foundation for the ThRAX2 regulatory mechanism of Cd stress. This study provides a genetic and theoretical basis for the bioremediation of Cd-contaminated land by cultivating transgenic plants in the future.

ThASR3 confers salt and osmotic stress tolerances in transgenic Tamarix and Arabidopsis.

BACKGROUND: ASR (abscisic acid-, stress-, and ripening-induced) gene family plays a crucial role in responding to abiotic stresses in plants. However, the roles of ASR genes protecting plants against high salt and drought stresses remain unknown in Tamarix hispida. RESULTS: In this study, a salt and drought-induced ASR gene, ThASR3, was isolated from Tamarix hispida. Transgenic Arabidopsis overexpressing ThASR3 exhibited stimulating root growth and increasing fresh weight compared with wild-type (WT) plants under both salt and water deficit stresses. To further analyze the gain- and loss-of-function of ThASR3, the transgenic T. hispida plants overexpressing or RNA interference (RNAi)-silencing ThASR3 were generated using transient transformation. The overexpression of ThASR3 in Tamarix and Arabidopsis plants displayed enhanced reactive oxygen species (ROS) scavenging capability under high salt and osmotic stress conditions, including increasing the activities of antioxidant enzymes and the contents of proline and betaine, and reducing malondialdehyde (MDA) content and electrolyte leakage rates. CONCLUSION: Our results indicate that ThASR3 functions as a positive regulator in Tamarix responses to salt and osmotic stresses and confers multiple abiotic stress tolerances in transgenic plants, which may have an important application value in the genetic improvement of forest tree resistance.

Tamarix hispida NAC Transcription Factor ThNAC4 Confers Salt and Drought Stress Tolerance to Transgenic Tamarix and Arabidopsis.

Salt and drought are considered two major abiotic stresses that have a significant impact on plants. Plant NAC (NAM, ATAF1/2, and CUC2) transcription factors (TFs) have been shown to play vital roles in plant development and responses to various abiotic stresses. ThNAC4, a NAC gene from Tamarix hispida involved in salt and osmotic stress tolerance, was identified and characterized in this study. According to a phylogenetic study, ThNAC4 is a member of NAC subfamily II. Subcellular localization analysis showed that ThNAC4 is located in the nucleus, and transcriptional activation experiments demonstrated that ThNAC4 is a transcriptional activator. Transgenic Arabidopsis plants overexpressing ThNAC4 exhibited improved salt and osmotic tolerance, as demonstrated by improved physiological traits. ThNAC4-overexpressing and ThNAC4-silenced T. hispida plants were generated using the transient transformation method and selected for gain- and loss-of-function analysis. The results showed that overexpression of ThNAC4 in transgenic Tamarix and Arabidopsis plants increased the activities of antioxidant enzymes (SOD, POD, and GST) and osmoprotectant (proline and trehalose) contents under stress conditions. These findings suggest that ThNAC4 plays an important physiological role in plant abiotic stress tolerance by increasing ROS scavenging ability and improving osmotic potential.

Transcriptomic Analysis of Cadmium Stressed Tamarix hispida Revealed Novel Transcripts and the Importance of Abscisic Acid Network.

Cadmium (Cd) pollution is widely detected in soil and has been recognized as a major environmental problem. Tamarix hispida is a woody halophyte, which can form natural forest on the desert and soil with 0.5 to 1% salt content, making it an ideal plant for the research on response to abiotic stresses. However, no systematic study has investigated the molecular mechanism of Cd tolerance in T. hispida. In the study, RNA-seq technique was applied to analyze the transcriptomic changes in T. hispida treated with 150 µmol L-1 CdCl2 for 24, 48, and 72 h compared with control. In total, 72,764 unigenes exhibited similar sequences in the Non-redundant nucleic acid database (NR database), while 36.3% of all these unigenes may be new transcripts. In addition, 6,778, 8,282, and 8,601 DEGs were detected at 24, 48, and 72 h, respectively. Functional annotation analysis indicated that many genes may be involved in Cd stress response, including ion bonding, signal transduction, stress sensing, hormone responses and ROS metabolism. A ThUGT gene from the abscisic acid (ABA) signaling pathway can enhance Cd resistance ability of T. hispida by regulating the production of ROS under Cd stress and inhibit absorption of Cd. The new transcriptome resources and data that we present in this study for T. hispida may facilitate investigation of molecular mechanisms governing Cd resistance.

Combined transcriptomic and metabolomic analysis reveals the potential mechanism of seed germination and young seedling growth in Tamarix hispida.

BACKGROUND: Seed germination is a series of ordered physiological and morphogenetic processes and a critical stage in plant life cycle. Tamarix hispida is one of the most salt-tolerant plant species; however, its seed germination has not been analysed using combined transcriptomics and metabolomics. RESULTS: Transcriptomic sequencing and widely targeted metabolomics were used to detect the transcriptional metabolic profiles of T. hispida at different stages of seed germination and young seedling growth. Transcriptomics showed that 46,538 genes were significantly altered throughout the studied development period. Enrichment study revealed that plant hormones, such as auxin, ABA, JA and SA played differential roles at varying stages of seed germination and post-germination. Metabolomics detected 1022 metabolites, with flavonoids accounting for the highest proportion of differential metabolites. Combined analysis indicated that flavonoid biosynthesis in young seedling growth, such as rhoifolin and quercetin, may improve the plant's adaptative ability to extreme desert environments. CONCLUSIONS: The differential regulation of plant hormones and the accumulation of flavonoids may be important for the seed germination survival of T. hispida in response to salt or arid deserts. This study enhanced the understanding of the overall mechanism in seed germination and post-germination. The results provide guidance for the ecological value and young seedling growth of T. hispida.

ThCOL2 Improves the Salt Stress Tolerance of Tamarix hispida.

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 COL genes with excellent salt tolerance from the woody halophyte Tamarix hispida, 8 ThCOL 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 ThCOL 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 T. hispida. In particular, the expression level of ThCOL2 changed significantly at most time points in the roots and leaves of T. hispida 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-ThCOL2 and pFGC5941-ThCOL2 were constructed for the transient transformation of T. hispida, and the transient infection of T. hispida with the pROKII empty vector was used as the control to further verify whether the ThCOL2 gene was involved in the regulation of the salt tolerance response of T. hispida. Overexpression of the ThCOL2 gene in plants under 150 mM NaCl stress increased the ability of transgenic T. hispida cells to remove reactive oxygen species (ROS) by regulating the activity of protective enzymes and promoting a decrease in the accumulation of O2- and H2O2, thereby reducing cell damage or cell death and enhancing salt tolerance. The ThCOL2 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 ThNCED1 and ThNCED4 were significantly higher, and the expressions of ThNCED3, ThZEP, and ThAAO3 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 ThCOL2 transcription factor will be searched and identified to analyze the salt tolerance regulatory network of ThCOL2.

ThHSFA1 Confers Salt Stress Tolerance through Modulation of Reactive Oxygen Species Scavenging by Directly Regulating ThWRKY4.

Heat shock transcription factors (HSFs) play critical roles in several types of environmental stresses. However, the detailed regulatory mechanisms in response to salt stress are still largely unknown. In this study, we examined the salt-induced transcriptional responses of ThHSFA1-ThWRKY4 in Tamarix hispida and their functions and regulatory mechanisms in salt tolerance. ThHSFA1 protein acts as an upstream regulator that can directly activate ThWRKY4 expression by binding to the heat shock element (HSE) of the ThWRKY4 promoter using yeast one-hybrid (Y1H), chromatin immunoprecipitation (ChIP), and dual-luciferase reporter assays. ThHSFA1 and ThWRKY4 expression was significantly induced by salt stress and abscisic acid (ABA) treatment in the roots and leaves of T. hispida. ThHSFA1 is a nuclear-localized protein with transactivation activity at the C-terminus. Compared to nontransgenic plants, transgenic plants overexpressing ThHSFA1 displayed enhanced salt tolerance and exhibited reduced reactive oxygen species (ROS) levels and increased antioxidant enzyme activity levels under salt stress. Therefore, we further concluded that ThHSFA1 mediated the regulation of ThWRKY4 in response to salt stress in T. hispida.

Comprehensive analysis of the stress associated protein (SAP) gene family in Tamarix hispida and the function of ThSAP6 in salt tolerance.

Stress associated proteins (SAPs), a class of A20/AN1 zinc finger domain-containing proteins, are involved in a variety of biotic and abiotic stress responses in plants. However, little is known about the SAP gene family and their functions in Tamarix hispida. In this study, we isolated and characterized 11 SAPs from T. hispida. The expression patterns of ThSAPs were analyzed under various stresses (salt and drought) and phytohormone treatment (SA, ABA and MeJA) using real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR). Most ThSAPs exhibited transcriptional responses to abiotic stresses and phytohormones. Among these ThSAPs, ThSAP6 was significantly induced by salt stress. Gain-and loss-of-function analyses revealed that ThSAP6 was a positive regulator of salt stress response. Overexpression of ThSAP6 in T. hispida increased antioxidant enzymes activity and proline content and decreased reactive oxygen species (ROS) accumulation and cell membrane damage under salt stress, while the opposite physiological changes were observed in ThSAP6-RNAi (RNA interference) lines. This study provides a comprehensive description of the SAP gene family in T. hispida, and demonstrates that ThSAP6 is a potential candidate for biotechnological approaches to improve salt tolerance in plants.

Revealing the salt tolerance mechanism of Tamarix hispida by large-scale identification of genes conferring salt tolerance.

The identification of genes conferring salt tolerance is important to reveal plant salt tolerance mechanisms. Here, we employed yeast expression system combined with high-throughput sequencing to identify genes conferring salt tolerance from Tamarix hispida Willd. A total of 1224 potential genes conferring salt tolerance were identified. Twenty-one genes were randomly selected for functional characterization using transient transformation in T. hispida and stable transformation in Arabidopsis thaliana (L.) Heynh. More than 90% of studied genes are found to confer tolerance to salt stress, indicating that the identified genes are reliable. More than 75% of the identified genes were highly expressed in roots rather than in leaves, suggesting roots play an important role in salt tolerance. The genes belonging to 'response to stimulus' were highly accumulated , and these accounted for 32% of the total identified genes. In addition, the processes of 'protein translation', 'osmotic adjustment', 'scavenging of free radicals', 'photosynthesis, detoxification of cells', 'protection of cellular macromolecules' and 'maintenance of cellular pH' play important roles in salt tolerance. This study provides useful information on the salt tolerance mechanism of T. hispida and offers a valuable resource for exploring genes used in salt tolerance breeding.

Overexpression of ThMYB8 mediates salt stress tolerance by directly activating stress-responsive gene expression.

MYB transcription factors are important in abiotic stress responses; however, the detailed mechanisms are unclear. Tamarix hispida contains multiple MYB genes. The present study characterized T. hispida MYB8 (ThMYB8) during salt stress using transgenic T. hispida and Arabidopsis assays. ThMYB8 overexpression and ThMYB8 RNAi analysis demonstrated that ThMYB8 enhanced the salt stress tolerance. Transgenic Arabidopsis ectopic expression of ThMYB8 significantly increased root growth, fresh weight, and seed germination rate compared with that of the wild-type under salt stress. Physiological parameters analysis in T. hispida and Arabidopsis showed that ThMYB8 overexpressing plants had the lowest levels of O2, H2O2, cell death, malondialdehyde, and electrolyte leakage. Overexpression of ThMYB8 regulated Na+ and K+ concentrations in plant tissues while maintaining K+/Na+ homeostasis. Analysis using qRT-PCR and ChIP-PCR identified possible downstream ThMYB8-regulated genes. ThMYB8 regulated the expression of ThCYP450-2 (cytochrome p450-2), Thltk (leucine-rich repeat transmembrane protein kinase), and ThTIP (aquaporin TIP) by binding to the MBSI motif ('CAACTG') in their promoters. The results indicated that ThMYB8 enhanced salt stress tolerance in T. hispida by regulating gene expression related to the activation of stress-associated physiological changes, such as enhanced reactive oxygen species scavenging capability, maintaining K+/Na+ homeostasis, and decreasing the malondialdehyde content and lipid peroxidation cell membranes.

Vacuolar membrane H+-ATPase c`` subunit gene (ThVHAc``1) from Tamarix hispida Willd improves salt stress tolerance.

The plant vacuolar H+-ATPase (V-ATPase) is a multisubunit complex. In addition to performing basic housekeeping functions, this complex is also involved in abiotic stress resistance in plants. In this study, a V-ATPase c`` subunit gene (ThVHAc``1) from Tamarix hispida Willd was cloned with a 534-bp ORF. Sequence analysis showed that the ThVHAc``1 protein contains four transmembrane helices and lacks a signal peptide. qRT-PCR results showed that ThVHAc``1 was primarily induced by treatments of NaCl, NaHCO3, PEG6000, CdCl2 or ABA in roots, stems and leaves of T. hispida. The expression pattern of ThVHAc``1 was significantly different from that of ThVHAc1 (a V-ATPase c subunit in T. hispida). Furthermore, the cell survival rates and density (OD600) results showed that the transgenic yeast overexpressing ThVHAc``1 exhibited increased tolerance to the above-mentioned abiotic stresses. In addition, the overexpression of ThVHAc``1 confers salt tolerance to transgenic Arabidopsis plants by improving the ROS content and decreasing the accumulation of O2- and H2O2. Similarly, the homologous transformation of the ThVHAc``1 gene into T. hispida also improved salt tolerance. Our results suggest that the ThVHAc``1 gene plays an important role in plant stress tolerance.

A 2-Cys peroxiredoxin gene from Tamarix hispida improved salt stress tolerance in plants.

BACKGROUND: Peroxiredoxins (Prxs) are a large family of antioxidant enzymes that respond to biotic and abiotic stress by decomposing reactive oxygen species (ROS). In this study, the stress tolerance function of the Th2CysPrx gene was further analysed. It lays a foundation for further studies on the salt tolerance molecular mechanism of T. hispida and improved salt tolerance via transgenic plants. RESULTS: In this study, the stress tolerance function of the Th2CysPrx gene was further analysed. The results of transgenic tobacco showed higher seed germination rates, root lengths, and fresh weight under salt stress than wild-type tobacco. Simultaneously, physiological indicators of transgenic tobacco and T. hispida showed that Th2CysPrx improved the activities of antioxidant enzymes and enhanced ROS removal ability to decrease cellular damage under salt stress. Moreover, Th2CysPrx improved the expression levels of four antioxidant genes (ThGSTZ1, ThGPX, ThSOD and ThPOD). CONCLUSIONS: Overall, these results suggested that Th2CysPrx enhanced the salt tolerance of the transgenic plants. These findings lay a foundation for further studies on the salt tolerance molecular mechanism of T. hispida and improved salt tolerance via transgenic plants.

Overexpression of ThSAP30BP from Tamarix hispida improves salt tolerance.

Histone deacetylases (HDACs) play an important regulatory role in plant response to biotic and abiotic stresses. They improve plant stress resistance by increasing the degree of histone acetylation associated with stress-responsive genes. SAP30BP, a human transcriptional regulatory protein, can increase histone deacetylase activity by regulating the deacetylation levels of lysines 9 and 14 in histone H3. In this study, a ThSAP30BP gene was cloned and characterized from Tamarix hispida (a kind of woody halophyte). The expression patterns of ThSAP30BP under different abiotic stresses and hormone treatments were detected by qRT-PCR. The results showed that ThSAP30BP was significantly upregulated at most time points under various stress treatments, suggesting that ThSAP30BP may be related to the abiotic stress response of T. hispida. To further analyze the salt stress resistance function of the ThSAP30BP gene, the plant overexpression vector pROKII-ThSAP30BP was instantaneously constructed and transformed into T. hispida. Meanwhile, the empty vector pROKII was also transformed as a control. The activities of SOD and POD, the contents of H2O2 and MDA, the relative conductance and the staining of NBT, DAB and Evans blue were analyzed and compared under salt stress. The results showed that the overexpression of ThSAP30BP in T. hispida reduced the accumulation of ROS in plants and the cell death rate under salt stress. These results suggested that ThSAP30BP may play an important physiological role in salt tolerance of T. hispida.

The ThSOS3 Gene Improves the Salt Tolerance of Transgenic Tamarix hispida and Arabidopsis thaliana.

The salt overly sensitive (SOS) signal transduction pathway is one of the most highly studied salt tolerance pathways in plants. However, the molecular mechanism of the salt stress response in Tamarix hispida has remained largely unclear. In this study, five SOS genes (ThSOS1-ThSOS5) from T. hispida were cloned and characterized. The expression levels of most ThSOS genes significantly changed after NaCl, PEG6000, and abscisic acid (ABA) treatment in at least one organ. Notably, the expression of ThSOS3 was significantly downregulated after 6 h under salt stress. To further analyze ThSOS3 function, ThSOS3 overexpression and RNAi-mediated silencing were performed using a transient transformation system. Compared with controls, ThSOS3-overexpressing transgenic T. hispida plants exhibited greater reactive oxygen species (ROS)-scavenging capability and antioxidant enzyme activity, lower malondialdehyde (MDA) and H2O2 levels, and lower electrolyte leakage rates under salt stress. Similar results were obtained for physiological parameters in transgenic Arabidopsis, including H2O2 and MDA accumulation, superoxide dismutase (SOD) and peroxidase (POD) activity, and electrolyte leakage. In addition, transgenic Arabidopsis plants overexpressing ThSOS3 displayed increased root growth and fresh weight gain under salt stress. Together, these data suggest that overexpression of ThSOS3 confers salt stress tolerance on plants by enhancing antioxidant enzyme activity, improving ROS-scavenging capability, and decreasing the MDA content and lipid peroxidation of cell membranes. These results suggest that ThSOS3 might play an important physiological role in salt tolerance in transgenic T. hispida plants. This study provides a foundation for further elucidation of salt tolerance mechanisms involving ThSOSs in T. hispida.

The NAC Protein from Tamarix hispida, ThNAC7, Confers Salt and Osmotic Stress Tolerance by Increasing Reactive Oxygen Species Scavenging Capability.

Plant specific NAC (NAM, ATAF1/2 and CUC2) transcription factors (TFs) play important roles in response to abiotic stress. In this study, we identified and characterized a NAC protein, ThNAC7, from Tamarix hispida. ThNAC7 is a nuclear localized protein and has transcriptional activation activity. ThNAC7 expression was markedly induced by salt and osmotic stresses. Transiently transformed T. hispida seedlings overexpressing ThNAC7 (OE) or with RNA interference (RNAi) silenced ThNAC7 were generated to investigate abiotic stress tolerance via the gain- and loss- of function. Overexpressing ThNAC7 showed an increased reactive oxygen species (ROS) scavenging capabilities and proline content, which was accomplished by enhancing the activities of superoxide dismutase (SOD) and peroxidase (POD) in transiently transformed T. hispida and stably transformed Arabidopsis plants. Additionally, ThNAC7 activated these physiological changes by regulating the transcription level of P5CS, SOD and POD genes. RNA-sequencing (RNA-seq) comparison between wild-type and ThNAC7-transformed Arabidopsis showed that more than 40 known salt tolerance genes might regulated by ThNAC7, including stress tolerance-related genes and TF genes. The results indicated that ThNAC7 induces the transcription level of genes associated with stress tolerance to enhance salt and osmotic stress tolerance via an increase in osmotic potential and enhanced ROS scavenging.