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18 pages, 6303 KiB

Open AccessArticle

Genome-Wide Identification of the 14-3-3 Gene Family and Its Involvement in Salt Stress Response through Interaction with NsVP1 in Nitraria sibirica Pall

by Xihong Wan, Rongfeng Duan, Huaxin Zhang, Jianfeng Zhu, Haiwen Wu, Huilong Zhang and Xiuyan Yang

Int. J. Mol. Sci. 2024, 25(6), 3432; https://doi.org/10.3390/ijms25063432 - 19 Mar 2024

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Abstract

14-3-3 proteins are widely distributed in eukaryotic cells and play an important role in plant growth, development, and stress tolerance. This study revealed nine 14-3-3 genes from the genome of Nitraria sibirica Pall., a halophyte with strong salt tolerance. The physicochemical properties, multiple [...] Read more.

14-3-3 proteins are widely distributed in eukaryotic cells and play an important role in plant growth, development, and stress tolerance. This study revealed nine 14-3-3 genes from the genome of Nitraria sibirica Pall., a halophyte with strong salt tolerance. The physicochemical properties, multiple sequence alignment, gene structure and motif analysis, and chromosomal distributions were analyzed, and phylogenetic analysis, cis-regulatory elements analysis, and gene transcription and expression analysis of Ns14-3-3s were conducted. The results revealed that the Ns14-3-3 gene family consists of nine members, which are divided into two groups: ε (four members) and non-ε (five members). These members are acidic hydrophilic proteins. The genes are distributed randomly on chromosomes, and the number of introns varies widely among the two groups. However, all genes have similar conserved domains and three-dimensional protein structures. The main differences are found at the N-terminus and C-terminus. The promoter region of Ns14-3-3s contains multiple cis-acting elements related to light, plant hormones, and abiotic stress responses. Transcriptional profiling and gene expression pattern analysis revealed that Ns14-3-3s were expressed in all tissues, although with varying patterns. Under salt stress conditions, Ns14-3-3 1a, Ns14-3-3 1b, Ns14-3-3 5a, and Ns14-3-3 7a showed significant changes in gene expression. Ns14-3-3 1a expression decreased in all tissues, Ns14-3-3 7a expression decreased by 60% to 71% in roots, and Ns14-3-3 1b expression increased by 209% to 251% in stems. The most significant change was observed in Ns14-3-3 5a, with its expression in stems increasing by 213% to 681%. The yeast two-hybrid experiments demonstrated that Ns14-3-3 5a interacts with NsVP1 (vacuolar H⁺-pyrophosphatase). This result indicates that Ns14-3-3 5a may respond to salt stress by promoting ionic vacuole compartmentalization in stems and leaves through interactions with NsVP1. In addition, N. sibirica has a high number of stems, allowing it to compartmentalize more ions through its stem and leaf. This may be a contributing factor to its superior salt tolerance compared to other plants. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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22 pages, 3653 KiB

Open AccessArticle

Populus euphratica GRP2 Interacts with Target mRNAs to Negatively Regulate Salt Tolerance by Interfering with Photosynthesis, Na⁺, and ROS Homeostasis

by Jing Li, Rui Zhao, Jian Liu, Jun Yao, Siyuan Ma, Kexin Yin, Ying Zhang, Zhe Liu, Caixia Yan, Nan Zhao, Xiaoyang Zhou and Shaoliang Chen

Int. J. Mol. Sci. 2024, 25(4), 2046; https://doi.org/10.3390/ijms25042046 - 07 Feb 2024

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Abstract

The transcription of glycine-rich RNA-binding protein 2 (PeGRP2) transiently increased in the roots and shoots of Populus euphratica (a salt-resistant poplar) upon initial salt exposure and tended to decrease after long-term NaCl stress (100 mM, 12 days). PeGRP2 overexpression in the [...] Read more.

The transcription of glycine-rich RNA-binding protein 2 (PeGRP2) transiently increased in the roots and shoots of Populus euphratica (a salt-resistant poplar) upon initial salt exposure and tended to decrease after long-term NaCl stress (100 mM, 12 days). PeGRP2 overexpression in the hybrid Populus tremula × P. alba ‘717-1B4’ (P. × canescens) increased its salt sensitivity, which was reflected in the plant’s growth and photosynthesis. PeGRP2 contains a conserved RNA recognition motif domain at the N-terminus, and RNA affinity purification (RAP) sequencing was developed to enrich the target mRNAs that physically interacted with PeGRP2 in P. × canescens. RAP sequencing combined with RT-qPCR revealed that NaCl decreased the transcripts of PeGRP2-interacting mRNAs encoding photosynthetic proteins, antioxidative enzymes, ATPases, and Na⁺/H⁺ antiporters in this transgenic poplar. Specifically, PeGRP2 negatively affected the stability of the target mRNAs encoding the photosynthetic proteins PETC and RBCMT; antioxidant enzymes SOD[Mn], CDSP32, and CYB1-2; ATPases AHA11, ACA8, and ACA9; and the Na⁺/H⁺ antiporter NHA1. This resulted in (i) a greater reduction in Fv/Fm, YII, ETR, and Pn; (ii) less pronounced activation of antioxidative enzymes; and (iii) a reduced ability to maintain Na⁺ homeostasis in the transgenic poplars during long-term salt stress, leading to their lowered ability to tolerate salinity stress. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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16 pages, 842 KiB

Open AccessArticle

Evolutionary Analysis of Six Gene Families Part of the Reactive Oxygen Species (ROS) Gene Network in Three Brassicaceae Species

by Thomas Horst Berthelier, Sébastien Christophe Cabanac, Caroline Callot, Arnaud Bellec, Catherine Mathé, Elisabeth Jamet and Christophe Dunand

Int. J. Mol. Sci. 2024, 25(3), 1938; https://doi.org/10.3390/ijms25031938 - 05 Feb 2024

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Abstract

Climate change is expected to intensify the occurrence of abiotic stress in plants, such as hypoxia and salt stresses, leading to the production of reactive oxygen species (ROS), which need to be effectively managed by various oxido-reductases encoded by the so-called ROS gene [...] Read more.

Climate change is expected to intensify the occurrence of abiotic stress in plants, such as hypoxia and salt stresses, leading to the production of reactive oxygen species (ROS), which need to be effectively managed by various oxido-reductases encoded by the so-called ROS gene network. Here, we studied six oxido-reductases families in three Brassicaceae species, Arabidopsis thaliana as well as Nasturtium officinale and Eutrema salsugineum, which are adapted to hypoxia and salt stress, respectively. Using available and new genomic data, we performed a phylogenomic analysis and compared RNA-seq data to study genomic and transcriptomic adaptations. This comprehensive approach allowed for the gaining of insights into the impact of the adaptation to saline or hypoxia conditions on genome organization (gene gains and losses) and transcriptional regulation. Notably, the comparison of the N. officinale and E. salsugineum genomes to that of A. thaliana highlighted changes in the distribution of ohnologs and homologs, particularly affecting class III peroxidase genes (CIII Prxs). These changes were specific to each gene, to gene families subjected to duplication events and to each species, suggesting distinct evolutionary responses. The analysis of transcriptomic data has allowed for the identification of genes related to stress responses in A. thaliana, and, conversely, to adaptation in N. officinale and E. salsugineum. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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21 pages, 6917 KiB

Open AccessArticle

Physiological, Epigenetic, and Transcriptome Analyses Provide Insights into the Responses of Wheat Seedling Leaves to Different Water Depths under Flooding Conditions

by Bo Li, Wei Hua, Shuo Zhang, Le Xu, Caixian Yang, Zhanwang Zhu, Ying Guo, Meixue Zhou, Chunhai Jiao and Yanhao Xu

Int. J. Mol. Sci. 2023, 24(23), 16785; https://doi.org/10.3390/ijms242316785 - 26 Nov 2023

Viewed by 985

Abstract

Flooding stress, including waterlogging and submergence, is one of the major abiotic stresses that seriously affects the growth and development of plants. In the present study, physiological, epigenetic, and transcriptomic analyses were performed in wheat seedling leaves under waterlogging (WL), half submergence (HS), [...] Read more.

Flooding stress, including waterlogging and submergence, is one of the major abiotic stresses that seriously affects the growth and development of plants. In the present study, physiological, epigenetic, and transcriptomic analyses were performed in wheat seedling leaves under waterlogging (WL), half submergence (HS), and full submergence (FS) treatments. The results demonstrate that FS increased the leaves’ hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) contents and reduced their chlorophyll contents (SPAD), photosynthetic efficiency (F_v/F_m), and shoot dry weight more than HS and WL. In addition, FS increased catalase (CAT) and peroxidase (POD) activities more than HS and WL. However, there were no significant differences in the contents of H₂O₂, MDA, SPAD, and F_v/F_m, and the activities of superoxide dismutase (SOD) and POD between the HS and WL treatments. The changes in DNA methylation were related to stress types, increasing under the WL and HS treatments and decreasing under the FS treatment. Additionally, a total of 9996, 10,619, and 24,949 genes were differentially expressed under the WL, HS, and FS treatments, respectively, among which the ‘photosynthesis’, ‘phenylpropanoid biosynthesis’, and ‘plant hormone signal transduction’ pathways were extensively enriched under the three flooding treatments. The genes involved in these pathways showed flooding-type-specific expression. Moreover, flooding-type-specific responses were observed in the three conditions, including the enrichment of specific TFs and response pathways. These results will contribute to a better understanding of the molecular mechanisms underlying the responses of wheat seedling leaves to flooding stress and provide valuable genetic and epigenetic information for breeding flood-tolerant varieties of wheat. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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14 pages, 3261 KiB

Open AccessArticle

Brevundimonas vesicularis (S1T13) Mitigates Drought-Stress-Associated Damage in Arabidopsis thaliana

by Can Thi My Tran, Tiba Nazar Ibrahim Al Azzawi, Murtaza Khan, Sajid Ali, Yong-Sun Moon and Byung-Wook Yun

Int. J. Mol. Sci. 2023, 24(23), 16590; https://doi.org/10.3390/ijms242316590 - 22 Nov 2023

Viewed by 677

Abstract

Drought stress is a significant threat to agricultural productivity and poses challenges to plant survival and growth. Research into microbial plant biostimulants faces difficulties in understanding complicated ecological dynamics, molecular mechanisms, and specificity; to address these knowledge gaps, collaborative efforts and innovative strategies [...] Read more.

Drought stress is a significant threat to agricultural productivity and poses challenges to plant survival and growth. Research into microbial plant biostimulants faces difficulties in understanding complicated ecological dynamics, molecular mechanisms, and specificity; to address these knowledge gaps, collaborative efforts and innovative strategies are needed. In the present study, we investigated the potential role of Brevundimonas vesicularis (S1T13) as a microbial plant biostimulant to enhance drought tolerance in Arabidopsis thaliana. We assessed the impact of S1T13 on Col-0 wild-type (WT) and atnced3 mutant plants under drought conditions. Our results revealed that the inoculation of S1T13 significantly contributed to plant vigor, with notable improvements observed in both genotypes. To elucidate the underlying mechanisms, we studied the role of ROS and their regulation by antioxidant genes and enzymes in plants inoculated with S1T13. Interestingly, the inoculation of S1T13 enhanced the activities of GSH, SOD, POD, and PPO by 33, 35, 41, and 44% in WT and 24, 22, 26, and 33% in atnced3, respectively. In addition, S1T13 upregulated the expression of antioxidant genes. This enhanced antioxidant machinery played a crucial role in neutralizing ROS and protecting plant cells from oxidative damage during drought stress. Furthermore, we investigated the impact of S1T13 on ABA and drought-stress-responsive genes. Similarly, S1T13 modulated the production of ABA and expression of AO3, ABA3, DREB1A, and DREB2A by 31, 42, 37, 41, and 42% in WT and 20, 29, 27, 38, and 29% in atnced3. The improvement in plant vigor, coupled with the induction of the antioxidant system and modulation of ABA, indicates the pivotal role of S1T13 in enhancing the drought stress tolerance of the plants. Conclusively, the current study provides valuable insights for the application of multitrait S1T13 as a novel strain to improve drought stress tolerance in plants and could be added to the consortium of biofertilizers. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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18 pages, 9713 KiB

Open AccessArticle

ZIP Genes Are Involved in the Retransfer of Zinc Ions during the Senescence of Zinc-Deficient Rice Leaves

by Yangming Ma, Yanfang Wen, Cheng Wang, Ziniu Wu, Xiaojuan Yuan, Ying Xiong, Kairui Chen, Limei He, Yue Zhang, Zhonglin Wang, Leilei Li, Zhiyuan Yang, Yongjian Sun, Zhongkui Chen and Jun Ma

Int. J. Mol. Sci. 2023, 24(18), 13989; https://doi.org/10.3390/ijms241813989 - 12 Sep 2023

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Abstract

Rice lacks sufficient amounts of zinc despite its vitality for human health. Leaf senescence enables redistribution of nutrients to other organs, yet Zn retransfer during deficiency is often overlooked. In this hydroponic experiment, we studied the effect of Zn deficiency on rice seedlings, [...] Read more.

Rice lacks sufficient amounts of zinc despite its vitality for human health. Leaf senescence enables redistribution of nutrients to other organs, yet Zn retransfer during deficiency is often overlooked. In this hydroponic experiment, we studied the effect of Zn deficiency on rice seedlings, focusing on the fourth leaf under control and deficient conditions. Growth phenotype analysis showed that the growth of rice nodal roots was inhibited in Zn deficiency, and the fourth leaf exhibited accelerated senescence and increased Zn ion transfer. Analyzing differentially expressed genes showed that Zn deficiency regulates more ZIP family genes involved in Zn ion retransfer. OsZIP3 upregulation under Zn-deficient conditions may not be induced by Zn deficiency, whereas OsZIP4 is only induced during Zn deficiency. Gene ontology enrichment analysis showed that Zn-deficient leaves mobilized more biological pathways (BPs) during aging, and the enrichment function differed from that of normal aging leaves. The most apparent “zinc ion transport” BP was stronger than that of normal senescence, possibly due to Zn-deficient leaves mobilizing large amounts of BP related to lipid metabolism during senescence. These results provide a basis for further functional analyses of genes and the study of trace element transfer during rice leaf senescence. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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19 pages, 10119 KiB

Open AccessArticle

The Over-Expression of Two R2R3-MYB Genes, PdMYB2R089 and PdMYB2R151, Increases the Drought-Resistant Capacity of Transgenic Arabidopsis

by Xueli Zhang, Haoran Wang, Ying Chen, Minren Huang and Sheng Zhu

Int. J. Mol. Sci. 2023, 24(17), 13466; https://doi.org/10.3390/ijms241713466 - 30 Aug 2023

Cited by 2 | Viewed by 1142

Abstract

The R2R3-MYB genes in plants play an essential role in the drought-responsive signaling pathway. Plenty of R2R3-MYB S21 and S22 subgroup genes in Arabidopsis have been implicated in dehydration conditions, yet few have been covered in terms of the role of the S21 [...] Read more.

The R2R3-MYB genes in plants play an essential role in the drought-responsive signaling pathway. Plenty of R2R3-MYB S21 and S22 subgroup genes in Arabidopsis have been implicated in dehydration conditions, yet few have been covered in terms of the role of the S21 and S22 subgroup genes in poplar under drought. PdMYB2R089 and PdMYB2R151 genes, respectively belonging to the S21 and S22 subgroups of NL895 (Populus deltoides × P. euramericana cv. ‘Nanlin895′), were selected based on the previous expression analysis of poplar R2R3-MYB genes that are responsive to dehydration. The regulatory functions of two target genes in plant responses to drought stress were studied and speculated through the genetic transformation of Arabidopsis thaliana. PdMYB2R089 and PdMYB2R151 could promote the closure of stomata in leaves, lessen the production of malondialdehyde (MDA), enhance the activity of the peroxidase (POD) enzyme, and shorten the life cycle of transgenic plants, in part owing to their similar conserved domains. Moreover, PdMYB2R089 could strengthen root length and lateral root growth. These results suggest that PdMYB2R089 and PdMYB2R151 genes might have the potential to improve drought adaptability in plants. In addition, PdMYB2R151 could significantly improve the seed germination rate of transgenic Arabidopsis, but PdMYB2R089 could not. This finding provides a clue for the subsequent functional dissection of S21 and S22 subgroup genes in poplar that is responsive to drought. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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19 pages, 10369 KiB

Open AccessArticle

Divergent Cross-Adaptation of Herbicide-Treated Wheat and Triticale Affected by Drought or Waterlogging

by Irina I. Vaseva, Margarita Petrakova, Ana Blagoeva and Dessislava Todorova

Int. J. Mol. Sci. 2023, 24(15), 12503; https://doi.org/10.3390/ijms241512503 - 06 Aug 2023

Cited by 1 | Viewed by 907

Abstract

Widely used agrochemicals that do not exert negative effects on crops and selectively target weeds could influence plant resilience under unfavorable conditions. The cross-adaptation of wheat (Triticum aestivum L.) and triticale (×Triticosecale Wittm.) exposed to two environmental abiotic stressors (drought and [...] Read more.

Widely used agrochemicals that do not exert negative effects on crops and selectively target weeds could influence plant resilience under unfavorable conditions. The cross-adaptation of wheat (Triticum aestivum L.) and triticale (×Triticosecale Wittm.) exposed to two environmental abiotic stressors (drought and waterlogging) was evaluated after treatment with a selective herbicide (Serrate^®, Syngenta). The ambivalent effects of the herbicide on the two studied crops were particularly distinct in waterlogged plants, showing a significant reduction in wheat growth and better performance of triticale individuals exposed to the same combined treatment. Histochemical staining for the detection of reactive oxygen species (ROS) confirmed that the herbicide treatment increased the accumulation of superoxide anion in the flooded wheat plants, and this effect persisted in the younger leaves of the recovered individuals. Comparative transcript profiling of ROS scavenging enzymes (superoxide dismutase, peroxidase, glutathione reductase, and catalase) in stressed and recovered plants revealed crop-specific variations resulting from the unfavorable water regimes in combination with the herbicide treatment. Short-term dehydration was relatively well tolerated by the hybrid crop triticale and this aligned with the considerable upregulation of genes for L-Proline biosynthesis. Its drought resilience was diminished by herbicide application, as evidenced by increased ROS accumulation after prolonged water deprivation. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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20 pages, 8016 KiB

Open AccessArticle

Transcriptome and Small RNA Sequencing Reveals the Basis of Response to Salinity, Alkalinity and Hypertonia in Quinoa (Chenopodium quinoa Willd.)

by Huanan Han, Yusen Qu, Yingcan Wang, Zaijie Zhang, Yuhu Geng, Yuanyuan Li, Qun Shao, Hui Zhang and Changle Ma

Int. J. Mol. Sci. 2023, 24(14), 11789; https://doi.org/10.3390/ijms241411789 - 22 Jul 2023

Cited by 1 | Viewed by 1011

Abstract

Quinoa (Chenopodium quinoa Willd.) is a dicotyledonous cereal that is rich in nutrients. This important crop has been shown to have significant tolerance to abiotic stresses such as salinization and drought. Understanding the underlying mechanism of stress response in quinoa would be [...] Read more.

Quinoa (Chenopodium quinoa Willd.) is a dicotyledonous cereal that is rich in nutrients. This important crop has been shown to have significant tolerance to abiotic stresses such as salinization and drought. Understanding the underlying mechanism of stress response in quinoa would be a significant advantage for breeding crops with stress tolerance. Here, we treated the low-altitude quinoa cultivar CM499 with either NaCl (200 mM), Na₂CO₃/NaHCO₃ (100 mM, pH 9.0) or PEG6000 (10%) to induce salinity, alkalinity and hypertonia, respectively, and analyzed the subsequent expression of genes and small RNAs via high-throughput sequencing. A list of known/novel genes were identified in quinoa, and the ones responding to different stresses were selected. The known/novel quinoa miRNAs were also identified, and the target genes of the stress response ones were predicted. Both the differently expressed genes and the targets of differently expressed miRNAs were found to be enriched for reactive oxygen species homeostasis, hormone signaling, cell wall synthesis, transcription factors and some other factors. Furthermore, we detected changes in reactive oxygen species accumulation, hormone (auxin and ethylene) responses and hemicellulose synthesis in quinoa seedlings treated with stresses, indicating their important roles in the response to saline, alkaline or hyperosmotic stresses in quinoa. Thus, our work provides useful information for understanding the mechanism of abiotic stress responses in quinoa, which would provide clues for improving breeding for quinoa and other crops. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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18 pages, 3261 KiB

Open AccessArticle

Comprehensive Analysis of Rice Seedling Transcriptome during Dehydration and Rehydration

by So Young Park and Dong-Hoon Jeong

Int. J. Mol. Sci. 2023, 24(9), 8439; https://doi.org/10.3390/ijms24098439 - 08 May 2023

Cited by 1 | Viewed by 1714

Abstract

Drought is a harmful abiotic stress that threatens the growth, development, and yield of rice plants. To cope with drought stress, plants have evolved their diverse and sophisticated stress-tolerance mechanisms by regulating gene expression. Previous genome-wide studies have revealed many rice drought stress-responsive [...] Read more.

Drought is a harmful abiotic stress that threatens the growth, development, and yield of rice plants. To cope with drought stress, plants have evolved their diverse and sophisticated stress-tolerance mechanisms by regulating gene expression. Previous genome-wide studies have revealed many rice drought stress-responsive genes that are involved in various forms of metabolism, hormone biosynthesis, and signaling pathways, and transcriptional regulation. However, little is known about the regulation of drought-responsive genes during rehydration after dehydration. In this study, we examined the dynamic gene expression patterns in rice seedling shoots during dehydration and rehydration using RNA-seq analysis. To investigate the transcriptome-wide rice gene expression patterns during dehydration and rehydration, RNA-seq libraries were sequenced and analyzed to identify differentially expressed genes (DEGs). DEGs were classified into five clusters based on their gene expression patterns. The clusters included drought-responsive DEGs that were either rapidly or slowly recovered to control levels by rehydration treatment. Representative DEGs were selected and validated using qRT-PCR. In addition, we performed a detailed analysis of DEGs involved in nitrogen metabolism, phytohormone signaling, and transcriptional regulation. In this study, we revealed that drought-responsive genes were dynamically regulated during rehydration. Moreover, our data showed the potential role of nitrogen metabolism and jasmonic acid signaling during the drought stress response. The transcriptome data in this study could be a useful resource for understanding drought stress responses in rice and provide a valuable gene list for developing drought-resistant crop plants. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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24 pages, 6814 KiB

Open AccessArticle

Populus euphratica GLABRA3 Binds PLDδ Promoters to Enhance Salt Tolerance

by Ying Zhang, Kexin Yin, Jun Yao, Ziyan Zhao, Zhe Liu, Caixia Yan, Yanli Zhang, Jian Liu, Jing Li, Nan Zhao, Rui Zhao, Xiaoyang Zhou and Shaoliang Chen

Int. J. Mol. Sci. 2023, 24(9), 8208; https://doi.org/10.3390/ijms24098208 - 03 May 2023

Cited by 4 | Viewed by 1845

Abstract

High NaCl (200 mM) increases the transcription of phospholipase Dδ (PLDδ) in roots and leaves of the salt-resistant woody species Populus euphratica. We isolated a 1138 bp promoter fragment upstream of the translation initiation codon of PePLDδ. A promoter–reporter construct, PePLDδ-pro [...] Read more.

High NaCl (200 mM) increases the transcription of phospholipase Dδ (PLDδ) in roots and leaves of the salt-resistant woody species Populus euphratica. We isolated a 1138 bp promoter fragment upstream of the translation initiation codon of PePLDδ. A promoter–reporter construct, PePLDδ-pro::GUS, was introduced into Arabidopsis plants (Arabidopsis thaliana) to demonstrate the NaCl-induced PePLDδ promoter activity in root and leaf tissues. Mass spectrometry analysis of DNA pull-down-enriched proteins in P. euphratica revealed that PeGLABRA3, a basic helix–loop–helix transcription factor, was the target transcription factor for binding the promoter region of PePLDδ. The PeGLABRA3 binding to PePLDδ-pro was further verified by virus-induced gene silencing, luciferase reporter assay (LRA), yeast one-hybrid assay, and electrophoretic mobility shift assay (EMSA). In addition, the PeGLABRA3 gene was cloned and overexpressed in Arabidopsis to determine the function of PeGLABRA3 in salt tolerance. PeGLABRA3-overexpressed Arabidopsis lines (OE1 and OE2) had a greater capacity to scavenge reactive oxygen species (ROS) and to extrude Na⁺ under salinity stress. Furthermore, the EMSA and LRA results confirmed that PeGLABRA3 interacted with the promoter of AtPLDδ in transgenic plants. The upregulated AtPLDδ in PeGLABRA3-transgenic lines resulted in an increase in phosphatidic acid species under no-salt and saline conditions. We conclude that PeGLABRA3 activated AtPLDδ transcription under salt stress by binding to the AtPLDδ promoter region, conferring Na⁺ and ROS homeostasis control via signaling pathways mediated by PLDδ and phosphatidic acid. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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21 pages, 10958 KiB

Open AccessArticle

A Cinnamate 4-HYDROXYLASE1 from Safflower Promotes Flavonoids Accumulation and Stimulates Antioxidant Defense System in Arabidopsis

by Yuying Hou, Yufei Wang, Xiaoyu Liu, Naveed Ahmad, Nan Wang, Libo Jin, Na Yao and Xiuming Liu

Int. J. Mol. Sci. 2023, 24(6), 5393; https://doi.org/10.3390/ijms24065393 - 11 Mar 2023

Cited by 3 | Viewed by 1330

Abstract

C4H (cinnamate 4-hydroxylase) is a pivotal gene in the phenylpropanoid pathway, which is involved in the regulation of flavonoids and lignin biosynthesis of plants. However, the molecular mechanism of C4H-induced antioxidant activity in safflower still remains to be elucidated. In this study, a [...] Read more.

C4H (cinnamate 4-hydroxylase) is a pivotal gene in the phenylpropanoid pathway, which is involved in the regulation of flavonoids and lignin biosynthesis of plants. However, the molecular mechanism of C4H-induced antioxidant activity in safflower still remains to be elucidated. In this study, a CtC4H1 gene was identified from safflower with combined analysis of transcriptome and functional characterization, regulating flavonoid biosynthesis and antioxidant defense system under drought stress in Arabidopsis. The expression level of CtC4H1 was shown to be differentially regulated in response to abiotic stresses; however, a significant increase was observed under drought exposure. The interaction between CtC4H1 and CtPAL1 was detected using a yeast two-hybrid assay and then verified using a bimolecular fluorescence complementation (BiFC) analysis. Phenotypic and statistical analysis of CtC4H1 overexpressed Arabidopsis demonstrated slightly wider leaves, long and early stem development as well as an increased level of total metabolite and anthocyanin contents. These findings imply that CtC4H1 may regulate plant development and defense systems in transgenic plants via specialized metabolism. Furthermore, transgenic Arabidopsis lines overexpressing CtC4H1 exhibited increased antioxidant activity as confirmed using a visible phenotype and different physiological indicators. In addition, the low accumulation of reactive oxygen species (ROS) in transgenic Arabidopsis exposed to drought conditions has confirmed the reduction of oxidative damage by stimulating the antioxidant defensive system, resulting in osmotic balance. Together, these findings have provided crucial insights into the functional role of CtC4H1 in regulating flavonoid biosynthesis and antioxidant defense system in safflower. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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21 pages, 5277 KiB

Open AccessArticle

Comprehensive Genome-Wide Analyses of Poplar R2R3-MYB Transcription Factors and Tissue-Specific Expression Patterns under Drought Stress

by Xueli Zhang, Haoran Wang, Ying Chen, Minren Huang and Sheng Zhu

Int. J. Mol. Sci. 2023, 24(6), 5389; https://doi.org/10.3390/ijms24065389 - 11 Mar 2023

Cited by 7 | Viewed by 2237

Abstract

R2R3-type MYB transcription factors are implicated in drought stress, which is a primary factor limiting the growth and development of woody plants. The identification of R2R3-MYB genes in the Populus trichocarpa genome has been previously reported. Nevertheless, the diversity and complexity of the [...] Read more.

R2R3-type MYB transcription factors are implicated in drought stress, which is a primary factor limiting the growth and development of woody plants. The identification of R2R3-MYB genes in the Populus trichocarpa genome has been previously reported. Nevertheless, the diversity and complexity of the conserved domain of the MYB gene caused inconsistencies in these identification results. There is still a lack of drought-responsive expression patterns and functional studies of R2R3-MYB transcription factors in Populus species. In this study, we identified a total of 210 R2R3-MYB genes in the P. trichocarpa genome, of which 207 genes were unevenly distributed across all 19 chromosomes. These poplar R2R3-MYB genes were phylogenetically divided into 23 subgroups. Collinear analysis demonstrated that the poplar R2R3-MYB genes underwent rapid expansion and that whole-genome duplication events were a dominant factor in the process of rapid gene expansion. Subcellular localization assays indicated that poplar R2R3-MYB TFs mainly played a transcriptional regulatory role in the nucleus. Ten R2R3-MYB genes were cloned from P. deltoides × P. euramericana cv. Nanlin895, and their expression patterns were tissue-specific. A majority of the genes showed similar drought-responsive expression patterns in two out of three tissues. This study provides a valid cue for further functional characterization of drought-responsive R2R3-MYB genes in poplar and provides support for the development of new poplar genotypes with elevated drought tolerance. Full article

(This article belongs to the Special Issue Plant Response to Abiotic Stress 2.0)

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