Positive Regulator In Response Research Articles

Genome-wide identification and expression pattern analysis of WRKY transcription factors in response to biotic and abiotic stresses in tea plants (Camellia sinensis)

Plants would encounter various biotic and abiotic stresses during the growth and development. WRKY transcription factors (TFs) as plant-specific TFs, play an important role in responding to various adverse circumstances. Despite some advances were achieved in functional studies of WRKY TFs in tea plants, systematic analysis of the involvement of CsWRKY TFs when facing cold, salt, drought stresses and pathogen and insect attack was lacked. In present study, a total of 78 CsWRKY TFs were identified following the genomic and transcript databases. The expression patterns of CsWRKYs in various organs of tea plants and the expression profiles in response to biotic and abiotic stresses were investigated by examining representative RNA-seq data. Moreover, the effects of hormone treatments (SA and MeJA) on the transcription levels of WRKY TFs were also investigated. The phylogenetic tree of CsWRKY TFs from different species indicated the functional diversity of WRKY TFs was not closely related to their protein classification. Concurrently, CsWRKY70-2 TF was identified as a positive regulator in response to drought stress. This study provided solid and valuable information, helping us better understand the functional diversity of CsWRKY TFs, and laid the foundation for further research on the function of key WRKY genes in tea plants.

BrERF109 positively regulates the tolerances of drought and salt stress in Chinese cabbage

Ethylene response factors (ERFs) are plant-specific transcription factors and play essential roles in plants’ defense responses to abiotic stresses. In this study, we identified an ethylene response factor BrERF109, and explored its roles in response to drought and salt stress. The BrERF109 protein contained an AP2 domain and was localized in the nucleus. Expression analysis suggested that BrERF109 was mainly expressed in root and leaf, and highly induced by drought, salt, heat, and cold stresses. Ectopic expression of BrERF109 in Arabidopsis improved the tolerance to drought and salt stress in transgenic plants, accompanied by reduced ABA sensitivity, enhanced antioxidant enzyme activities and chlorophyll content, as well as induced the expression of canonical stress responsive genes. Silencing of BrERF109 in Chinese cabbage by virus-induced gene silencing (VIGS) led to plants’ susceptibility to drought and salt stress, and suppressed superoxide dismutase and peroxidase activities. Furthermore, BrLTP3, BrCIPK6, BrAZF2, and BrZAT10 genes containing GCC-box in their promoter regions were identified as downstream candidate targets of BrERF109. Overall, our study demonstrated that BrERF109 is a key positive regulator in response to drought and salt stress in Chinese cabbage, and serves as a valuable target gene for breeding new cultivars with enhanced tolerances to abiotic stresses in Brassica vegetables.

Rice OsANN9 Enhances Drought Tolerance through Modulating ROS Scavenging Systems.

Drought is a critical abiotic stress which leads to crop yield and a decrease in quality. Annexins belong to a multi-gene family of calcium- and lipid-binding proteins and play diverse roles in plant growth and development. Herein, we report a rice annexin protein, OsANN9, which in addition to regular annexin repeats and type-II Ca2+ binding sites, also consists of a C2H2-type zinc-finger domain. We found that the expression of OsANN9 was upregulated by polyethylene glycol (PEG) or water-deficient treatment. Moreover, plants that overexpressed OsANN9 had increased survival rates under drought stress, while both OsANN9-RNAi and osann9 mutants showed sensitivity to drought. In addition, the overexpression of OsANN9 increased superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities, which regulate reactive oxygen species homeostasis. Collectively, these findings indicate that OsANN9 may function as a positive regulator in response to drought stress by modulating antioxidant accumulation. Interestingly, the setting rates of osann9 mutant rice plants significantly decreased in comparison to wild-type plants, suggesting that OsANN9 might be involved in other molecular mechanisms in the rice seed development stage.

Multiple roles of wheat ferritin genes during stress treatment and TaFER5D-1 as a positive regulator in response to drought and salt tolerance

Ferritin not only regulates the plant's iron content but also plays a significant role in the plant's development and resistance to oxidative damage. However, the role of the FER family in wheat has not been systematically elucidated. In this study, 39 FERs identified from wheat and its ancestral species were clustered into two subgroups, and gene members from the same group contain relatively conservative protein models. The structural analyses indicated that the gene members from the same group contained relatively conserved protein models. The cis-acting elements and expression patterns analysis suggested that TaFERs might play an important role combating to abiotic and biotic stresses. In the transcriptional analysis, the TaFER5D-1 gene was found to be significantly up-regulated under drought and salt stresses and was, therefore, selected to further explore the biological functions Moreover, the GFP expression assay revealed the subcellular localization of TaFER5D-1 proteins in the chloroplast, nucleus, membrane and cytoplasm. Over-expression of TaFER5D-1 in transgenic Arabidopsis lines conferred greater tolerance to drought and salt stress. According to the qRT-PCR data, TaFER5D-1 gene over-expression increased the expression of genes related to root development (Atsweet-17 and AtRSL4), iron storage (AtVIT1 and AtYSL1), and stress response (AtGolS1 and AtCOR47). So it is speculated that TaFER5D-1 could improve stress tolerance by promoting root growth, iron storage, and stress-response ability. Thus, the current study provides insight into the role of TaFER genes in wheat.

The JASMONATE ZIM-domain-OPEN STOMATA1 cascade integrates jasmonic acid and abscisic acid signaling to regulate drought tolerance by mediating stomatal closure in poplar.

Drought, which directly affects the yield of crops and trees, is a natural stress with a profound impact on the economy. Improving water use efficiency (WUE) and drought tolerance are relatively effective strategies to alleviate drought stress. OPEN STOMATA1 (OST1), at the core of abscisic acid (ABA) signaling, can improve WUE by regulating stomatal closure and photosynthesis. Methyl jasmonate (MeJA) and ABA crosstalk is considered to be involved in the response to drought stress, but the detailed molecular mechanism is insufficiently known. Here, Populus euphratica, which naturally grows in arid and semiarid regions, was selected as the species for studying MeJA and ABA crosstalk under drought. A yeast two-hybrid assay was performed using PeOST1 as bait and a nucleus-localized factor, JASMONATE ZIM-domain protein 2 (PeJAZ2), was found to participate in MeJA signaling by interacting with PeOST1. Overexpression of PeJAZ2 in poplar notably increased water deficit tolerance and WUE in both severe and mild drought stress by regulating ABA signaling rather than ABA synthesis. Furthermore, a PeJAZ2 overexpression line was shown to have greater ABA-induced stomatal closure and hydrogen peroxide (H2O2) production. Collectively, this evidence establishes a mechanism in which PeJAZ2 acts as a positive regulator in response to drought stress via ABA-induced stomatal closure caused by H2O2 production. Our study presents a new insight into the crosstalk of ABA and jasmonic acid signaling in regulating WUE and drought stress, providing a basis of the drought tolerance mechanism of P. euphratica.

Multiple roles of wheat calmodulin genes during stress treatment and TaCAM2-D as a positive regulator in response to drought and salt tolerance

Calmodulin (CaM) and calmodulin-like (CML) proteins are the most prominent calcium (Ca2+) sensing proteins involved in Ca2+-signaling processes. However, the function of these calcium sensors in wheat remains unclear. In this study, 15 TaCAMs and 113 TaCMLs were identified from the wheat reference genome. The analysis of cis-acting elements and expression patterns showed that TaCAMs might play an important role in response to abiotic and biotic stresses. TaCAM2-D gene was found to be significantly upregulated under drought and salt stresses, and thus, it was selected to further explore the biological function. Moreover, TaCAM2-D was observed to be localized in the nucleus, membrane and cytoplasm. Overexpression of TaCAM2-D in Arabidopsis conferred greater tolerance to drought and salt. The prediction analysis, the yeast two-hybrid analysis, and bimolecular fluorescence complementation assay indicated that TaCAM2-D interacted with TaMPK8, which is one of the wheat mitogen-activated protein kinases. Thus, the current study provides insights into the understanding of the TaCAM and TaCML genes in wheat.

A C2H2-type zinc finger transcription factor, MdZAT17, acts as a positive regulator in response to salt stress

Salt stress restricts plant growth and productivity worldwide. Zinc finger proteins play important roles in response to various abiotic plant stresses. In this research, we identified and characterized the ZAT17 gene in Malus domestica, which encodes a C2H2-type zinc finger protein. MdZAT17 has two typical conserved zinc finger domains and an ERF-associated amphiphilic repression (EAR) motif. Promoter analysis showed that MdZAT17 contains several stress-related response elements (ABRE, CGTCA-motif, and TC-rich repeats), and qRT-PCR analysis showed that the expression level of MdZAT17 was induced by various abiotic stress treatments. The overexpression of MdZAT17 improved tolerance to salt stress in apple calli. The ectopic expression of MdZAT17 in Arabidopsis enhanced salt stress tolerance and led to lower malondialdehyde (MDA) content, lower reactive oxygen species (ROS) accumulation, and greater anthocyanin accumulation under salt stress. Moreover, the overexpression of MdZAT17 transgenic apple calli and Arabidopsis reduced the sensitivity to abscisic acid (ABA). In conclusion, our results indicate that MdZAT17 plays a positive regulatory role in salt tolerance, providing a theoretical basis for further research on its molecular mechanisms.

Lilium regale Wilson WRKY3 modulates an antimicrobial peptide gene, LrDef1, during response to Fusarium oxysporum

BackgroundWRKY transcription factors (TFs) play vital roles in plant growth and development, secondary metabolite synthesis, and response to biotic and abiotic stresses. In a previous transcriptome sequencing analysis of Lilium regale Wilson, we identified multiple WRKY TFs that respond to exogenous methyl jasmonate treatment and lily Fusarium wilt (Fusarium oxysporum).ResultsIn the present study, the WRKY TF LrWRKY3 was further analyzed to reveal its function in defense response to F. oxysporum. The LrWRKY3 protein was localized in the plant cell nucleus, and LrWRKY3 transgenic tobacco lines showed higher resistance to F. oxysporum compared with wild-type (WT) tobacco. In addition, some genes related to jasmonic acid (JA) biosynthesis, salicylic acid (SA) signal transduction, and disease resistance had higher transcriptional levels in the LrWRKY3 transgenic tobacco lines than in the WT. On the contrary, L. regale scales transiently expressing LrWRKY3 RNA interference fragments showed higher sensitivity to F. oxysporum infection. Moreover, a F. oxysporum-induced defensin gene, Def1, was isolated from L. regale, and the recombinant protein LrDef1 isolated and purified from Escherichia coli possessed antifungal activity to several phytopathogens, including F. oxysporum. Furthermore, co-expression of LrWRKY3 and the LrDef1 promoter in tobacco enhanced the LrDef1 promoter-driven expression activity.ConclusionsThese results clearly indicate that LrWRKY3 is an important positive regulator in response to F. oxysporum infection, and one of its targets is the antimicrobial peptide gene LrDef1.

TIP60 buffers acute stress response and depressive behaviour by controlling PPARγ-mediated transcription

Tat-interacting protein 60 (TIP60) as nuclear receptors (NRs) coregulator, acts as a tumor suppressor and also has promising therapeutic potential to target Alzheimer’s disease. Stress has been implicated in many psychiatric disorders, and these disorders are characterized by impairments in cognitive function. Until now, there are no experimental data available on the regulatory effect of TIP60 in acute stress and depression. There is also no definitive explanation on which specific modulation of target gene expression is achieved by TIP60. Here, we identify TIP60 as a novel positive regulator in response to acute restraint stress (ARS) and a potentially effective target of antidepressants. Firstly, we discovered increased hippocampal TIP60 expressions in the ARS model. Furthermore, using the TIP60 inhibitor, MG149, we proved that TIP60 function correlates with behavioral and synaptic activation in the two-hour ARS. Secondly, the lentivirus vector (LV)-TIP60overexpression (OE) was injected into the hippocampus prior to the chronic restraint stress (CRS) experiments and it was found that over-expressed TIP60 compensates for TIP60 decrease and improves depression index in CRS. Thirdly, through the intervention of TIP60 expression in vitro, we established the genetic regulation of TIP60 on synaptic proteins, confirmed the TIP60 function as a specific coactivator for PPARγ and found that the PPARγ-mediated TIP60 function modulates transcriptional activation of synaptic proteins. Finally, the LV-TIP60OE and PPARγ antagonist, GW9662, were both administered in the CRS model and the data indicated that blocking PPARγ significantly weakened the protective effect of TIP60 against the CRS-induced depression. Conclusively, these findings together support TIP60 as a novel positive factor in response to acute stress and interacts with PPARγ to modulate the pathological mechanism of CRS-induced depression.

Silencing GhJUB1L1 (JUB1-like 1) reduces cotton (Gossypium hirsutum) drought tolerance.

Drought stress massively restricts plant growth and the yield of crops. Reducing the deleterious effects of drought is necessary for agricultural industry. The plant-specific NAC (NAM, ATAF1/2 and CUC2) transcription factors (TFs) are widely involved in the regulation of plant development and stress response. One of the NAC TF, JUNGBRUNNEN1 (JUB1), has been reported to involve in drought resistance in Arabidopsis. However, little is known of how the JUB1 gene respond to drought stress in cotton. In the present study, we cloned GhJUB1L1, a homologous gene of JUB1 in upland cotton. GhJUB1L1 is preferentially expressed in stem and leaf and could be induced by drought stress. GhJUB1L1 protein localizes to the cell nucleus, and the transcription activation region of which is located in the C-terminal region. Silencing GhJUB1L1 gene via VIGS () reduced cotton drought tolerance, and retarded secondary cell wall (SCW) development. Additionally, the expression of some drought stress-related genes and SCW synthesis-related genes were altered in the GhJUB1L1 silencing plants. Collectively, our findings indicate that GhJUB1L1 may act as a positive regulator in response to drought stress and SCW development in cotton. Our results enriched the roles of NAC TFs in cotton drought tolerance and laid a foundation for the cultivation of transgenic cotton with higher drought tolerance.

AaWRKY17, a positive regulator of artemisinin biosynthesis, is involved in resistance to Pseudomonas syringae in Artemisia annua

Artemisia annua, a traditional Chinese medicinal plant, remains the only plant source for artemisinin production, yet few genes have been identified to be involved in both the response to biotic stresses, such as pathogens, and artemisinin biosynthesis. Here, we isolated and identified the WRKY transcription factor (TF) AaWRKY17, which could significantly increase the artemisinin content and resistance to Pseudomonas syringae in A. annua. Yeast one-hybrid (Y1H), dual-luciferase (dual-LUC), and electrophoretic mobility shift assay (EMSA) results showed that AaWRKY17 directly bound to the W-box motifs in the promoter region of the artemisinin biosynthetic pathway gene amorpha-4,11-diene synthase (ADS) and promoted its expression. Real-time quantitative PCR (RT-qPCR) analysis revealed that the transcript levels of two defense marker genes, Pathogenesis-Related 5 (PR5) and NDR1/HIN1-LIKE 10 (NHL10), were greatly increased in AaWRKY17-overexpressing transgenic A. annua plants. Additionally, overexpression of AaWRKY17 in A. annua resulted in decreased susceptibility to P. syringae. These results indicated that AaWRKY17 acted as a positive regulator in response to P. syringae infection. Together, our findings demonstrated that the novel WRKY transcription factor AaWRKY17 could potentially be used in transgenic breeding to improve the content of artemisinin and pathogen tolerance in A. annua.

OsMADS23 phosphorylated by SAPK9 confers drought and salt tolerance by regulating ABA biosynthesis in rice.

Some of MADS-box transcription factors (TFs) have been shown to play essential roles in the adaptation of plant to abiotic stress. Still, the mechanisms that MADS-box proteins regulate plant stress response are not fully understood. Here, a stress-responsive MADS-box TF OsMADS23 from rice conferring the osmotic stress tolerance in plants is reported. Overexpression of OsMADS23 remarkably enhanced, but knockout of the gene greatly reduced the drought and salt tolerance in rice plants. Further, OsMADS23 was shown to promote the biosynthesis of endogenous ABA and proline by activating the transcription of target genes OsNCED2, OsNCED3, OsNCED4 and OsP5CR that are key components for ABA and proline biosynthesis, respectively. Then, the convincing evidence showed that the OsNCED2-knockout mutants had lower ABA levels and exhibited higher sensitivity to drought and oxidative stress than wild type, which is similar to osmads23 mutant. Interestingly, the SnRK2-type protein kinase SAPK9 was found to physically interact with and phosphorylate OsMADS23, and thus increase its stability and transcriptional activity. Furthermore, the activation of OsMADS23 by SAPK9-mediated phosphorylation is dependent on ABA in plants. Collectively, these findings establish a mechanism that OsMADS23 functions as a positive regulator in response to osmotic stress by regulating ABA biosynthesis, and provide a new strategy for improving drought and salt tolerance in rice.

The Pepper Mitogen-Activated Protein Kinase CaMAPK7 Acts as a Positive Regulator in Response to Ralstonia solanacearum Infection.

Mitogen-activated protein kinase (MAPK) pathways play a vital role in multiple plant processes, including growth, development, and stress signaling, but their involvement in response to Ralstonia solanacearum is poorly understood, particularly in pepper plants. Herein, CaMAPK7 was identified from the pepper genome and functionally analyzed. The accumulations of CaMAPK7 transcripts and promoter activities were both significantly induced in response to R. solanacearum strain FJC100301 infection, and exogenously applied phytohormones, including methyl jasmonate (MeJA), brassinolide (BR), salicylic acid (SA), and ethephon (ETN), were decreased by abscisic acid (ABA) treatment. Virus-induced gene silencing (VIGS) of CaMAPK7 significantly enhanced the susceptibility of pepper plants to infection by R. solanacearum and downregulated the defense-related marker genes, including CaDEF1, CaPO2, CaSAR82A, and CaWRKY40. In contrast, the ectopic overexpression of CaMAPK7 in transgenic tobacco enhanced resistance to R. solanacearum and upregulated the defense-associated marker genes, including NtHSR201, NtHSR203, NtPR4, PR1a/c, NtPR1b, NtCAT1, and NtACC. Furthermore, transient overexpression of CaMAPK7 in pepper leaves triggered intensive hypersensitive response (HR)-like cell death, H2O2 accumulation, and enriched CaWRKY40 at the promoters of its target genes and drove their transcript accumulations, including CaDEF1, CaPO2, and CaSAR82A. Taken together, these data indicate that R. solanacearum infection induced the expression of CaMAPK7, which indirectly modifies the binding of CaWRKY40 to its downstream targets, including CaDEF1, CaPO2, and CaSAR82A, ultimately leading to the activation of pepper immunity against R. solanacearum. The protein that responds to CaMAPK7 in pepper plants should be isolated in the future to build a signaling bridge between CaMAPK7 and CaWRKY40.

Oryza sativa, C4HC3-type really interesting new gene (RING), OsRFPv6, is a positive regulator in response to salt stress by regulating Na+ absorption.

Salinity negatively affects plant growth, productivity, and metabolism. Therefore, plants have evolved diverse strategies to survive in saline environments. To identify such strategies involving the ubiquitin/26S proteasome system, we characterized molecular functions of a rice C4HC3 really interesting new gene (RING)-type E3-ubiquitin ligase gene. Oryza sativa RING finger protein v6 (OsRFPv6) was highly expressed under conditions of abiotic stress, induced by 100 mM NaCl and 20% PEG. The GFP-OsRFPv6 protein was localized in the plasma membrane and cytosol in rice protoplasts. In vitro ubiquitin assay revealed that OsRFPv6 possessed E3-ubiquitin ligase activity, but its variant OsRFPv6C100A did not. OsRFPv6-overexpressing plants were insensitive to salinity, but their growth was delayed under normal conditions. Under saline conditions, transgenic plants exhibited higher proline, soluble sugar, and chlorophyll content and lower H2 O2 accumulation than wild-type plants. Moreover, transgenic plants exhibited lower Na+ uptake, lower Na+ content, and higher K+ content in the xylem sap assay. Under saline conditions, the expression levels of nine Na+ /K+ transporter genes in roots and leaves were significantly different between transgenic and wild-type plants. Specifically, under both normal and saline conditions, the expression of OsHKT2;1, a Na+ transporter, in the roots of transgenic plants was lower than that in the roots of wild-type plants. These results suggest that OsRFPv6 E3-ubiquitin ligase serves as a positive regulator of salinity response via Na+ uptake.

MdCIB1, an apple bHLH transcription factor, plays a positive regulator in response to drought stress

Drought restricts the plant growth and productivity worldwide. The basic helix-loop-helix (bHLH) transcription factors play key roles in response to various abiotic stresses. In this study, we identified and characterized a bHLH transcription factor, cryptochrome-interacting bHLH 1 (MdCIB1), from apple (Malus domestica). MdCIB1 had a typical bHLH domain and was localized in the nucleus. Moreover, MdCIB1 also interacted with MdCRY2, and participated in blue light response and flowering time regulation. RT-qPCR analysis showed that PEG 6000 notably induced the transcript level of MdCIB1. MdCIB1-overexpressing apple calli displayed improved tolerance to PEG 6000 treatment. Ectopic expression of MdCIB1 in Arabidopsis improved drought stress resistance, as reflected by better physiological indexes under drought stress, such as longer roots, higher chlorophyll levels and survival rate, and lower water loss rate of leaves than Col control. Further investigation demonstrated that MdCIB1 played a positive regulator in ABA sensitivity and ABA-mediated stomatal closure. In addition, MdCIB1 transgenic Arabidopsis exhibited higher proline content, lower malondialdehyde (MDA) content, and H2O2 and O2− accumulation, as well as increased the activities of antioxidant enzymes under drought stress, which were able to protect plants from osmotic and oxidative damages. Besides, the transcripts of ROS scavenging-related genes and stress-responsive genes were also significantly upregulated in MdCIB1 transgenic lines. In summary, our study demonstrated that MdCIB1 plays a positive role in drought resistance.

Isolation and Functional Characterization of a Salt-Responsive Calmodulin-Like Gene MpCML40 from Semi-Mangrove Millettia pinnata.

Salt stress is a major increasing threat to global agriculture. Pongamia (Millettia pinnata), a semi-mangrove, is a good model to study the molecular mechanism of plant adaptation to the saline environment. Calcium signaling pathways play critical roles in the model plants such as Arabidopsis in responding to salt stress, but little is known about their function in Pongamia. Here, we have isolated and characterized a salt-responsive MpCML40, a calmodulin-like (CML) gene from Pongamia. MpCML40 protein has 140 amino acids and is homologous with Arabidopsis AtCML40. MpCML40 contains four EF-hand motifs and a bipartite NLS (Nuclear Localization Signal) and localizes both at the plasma membrane and in the nucleus. MpCML40 was highly induced after salt treatment, especially in Pongamia roots. Heterologous expression of MpCML40 in yeast cells improved their salt tolerance. The 35S::MpCML40 transgenic Arabidopsis highly enhanced seed germination rate and root length under salt and osmotic stresses. The transgenic plants had a higher level of proline and a lower level of MDA (malondialdehyde) under normal and stress conditions, which suggested that heterologous expression of MpCML40 contributed to proline accumulation to improve salt tolerance and protect plants from the ROS (reactive oxygen species) destructive effects. Furthermore, we did not observe any measurable discrepancies in the development and growth between the transgenic plants and wild-type plants under normal growth conditions. Our results suggest that MpCML40 is an important positive regulator in response to salt stress and of potential application in producing salt-tolerant crops.

Overexpression of DBF-Interactor Protein 6 Containing an R3H Domain Enhances Drought Tolerance in Populus L. (Populus tomentosa).

Drought is the primary disaster that endangers agricultural production, including animal husbandry, and affects the distribution, growth, yield, and quality of crops. Previous study had revealed that DIP, as a potential regulator of DBF activity, played an important role in response to drought stress in maize. In this study, a total of 67 DIPs were identified from seventeen land plants, including six tobacco DIPs (NtDIPs). NtDIP6 gene was further selected as a candidate gene for subsequent experiments based on the phylogenetic analysis and structural analysis. The transgenic tobacco and poplar plants over-expressing NtDIP6 gene were generated using the Agrobacterium- mediated method. Although there was not phenotypic difference between transgenic plants and wild-type plants under normal conditions, overexpression of the NtDIP6 gene in transgenic tobacco and poplar plants enhanced the drought tolerance under drought treatments in comparison with the wild type. The content of antioxidant defense enzymes peroxidase (POD), catalase (CAT), and the photosynthetic rate increased in NtDIP6-Ox transgenic tobacco and poplar plants, while the content of malondialdehyde decreased, suggesting that the overexpression of NtDIP6 enhances the antioxidant capacity of transgenic poplar. Furthermore, the results of qRT-PCR showed that the level of expression of drought-related response genes significantly increased in the NtDIP6-Ox transgenic plants. These results indicated that NtDIP6, as a positive response regulator, improves drought stress tolerance by scavenging superoxide via the accumulation of antioxidant defense enzymes.

Pepper CaMLO6 Negatively Regulates Ralstonia solanacearum Resistance and Positively Regulates High Temperature and High Humidity Responses.

Plant mildew-resistance locus O (MLO) proteins influence susceptibility to powdery mildew. However, their roles in plant responses to other pathogens and heat stress remain unclear. Here, we showed that CaMLO6, a pepper (Capsicum annuum) member of MLO clade V, is a protein targeted to plasma membrane and probably endoplasmic reticulum. The transcript expression level of CaMLO6 was upregulated in the roots and leaves of pepper plants challenged with high temperature and high humidity (HTHH) and was upregulated in leaves but downregulated in roots of plants infected with the bacterial pathogen Ralstonia solanacearum. CaMLO6 was also directly upregulated by CaWRKY40 upon HTHH but downregulated by CaWRKY40 upon R. solanacearum infection. Virus-induced gene silencing of CaMLO6 significantly decreased pepper HTHH tolerance and R. solanacearum susceptibility. Moreover, CaMLO6 overexpression enhanced the susceptibility of Nicotiana benthamiana and pepper plants to R. solanacearum and their tolerance to HTHH, effects that were associated with the expression of immunity- and thermotolerance-associated marker genes, respectively. These results suggest that CaMLO6 acts as a positive regulator in response to HTHH but a negative regulator in response to R. solanacearum. Moreover, CaMLO6 is transcriptionally affected by R. solanacearum and HTHH; these transcriptional responses are at least partially regulated by CaWRKY40.

SbWRKY30 enhances the drought tolerance of plants and regulates a drought stress-responsive gene, SbRD19, in sorghum

WRKY transcription factors have been suggested to play important roles in response and adaptation to drought stress. However, how sorghum WRKY transcription factors function in drought stress is still unclear. Here, we identify a WRKY transcription factor of sorghum, SbWRKY30, which is induced significantly by drought stress. SbWRKY30 is mainly expressed in sorghum taproot and leaf. SbWRKY30 has transcriptional activation activity and functions in the nucleus. Heterologous expression of SbWRKY30 confers tolerance to drought stress in Arabidopsis (Arabidopsis thaliana) and rice by affecting root architecture. In addition, SbWRKY30 transgenic Arabidopsis and rice plants have higher proline contents and SOD, POD, and CAT activities but lower MDA contents than wild-type plants after drought stress. As a homologous gene of the drought stress-responsive gene RD19 of Arabidopsis, SbRD19 overexpression in Arabidopsis improved the drought tolerance of plants relative to wild-type plants. Further analysis demonstrated that SbWRKY30 could induce SbRD19 expression through binding to the W-box element in the promoter of SbRD19. These results suggest that SbWRKY30 functions as a positive regulator in response to drought stress. Therefore, SbWRKY30 may serve as a promising candidate gene for molecular breeding to generate drought-tolerant crops.

Overexpression of a Multiprotein Bridging Factor 1 Gene DgMBF1 Improves the Salinity Tolerance of Chrysanthemum.

Soil salinity represents a major constraint in the growth of chrysanthemum. Therefore, improving salinity tolerance of chrysanthemum has become an important research direction in tolerance breeding. Multiprotein bridging factor 1 (MBF1) is an evolutionarily highly conserved transcriptional co-activator in archaea and eukaryotes and has been reported to play important roles to respond to abiotic stresses. Here, a MBF1 gene induced by salt stress was isolated and functionally characterized from Dendranthema grandiflorum and name as DgMBF1. Overexpression of DgMBF1 in chrysanthemum increased the tolerance of plants to high salt stress compared to wild type (WT). It also showed fewer accumulations of hydrogen peroxide (H2O2), superoxide anion (O2−), higher activities of antioxidant enzymes, more content of proline and soluble sugar (SS) and more favorable K+/Na+ ratio than those of WT under salt stress. In addition, the expression level of genes related to antioxidant biosynthesis, proline biosynthesis, glyco-metabolism and K+/Na+ homeostasis was statistically significant higher in the DgMBF1-overexpressed lines than that in WT. These results demonstrated that DgMBF1 is a positive regulator in response to salt stress and could serve as a new candidate gene for salt-tolerant plant breeding.