Cloning and molecular characterization of five MAPKK genes from grapevine (Vitis vinifera) and two of them positively regulate salt and drought tolerance in transgenic Arabidopsis

Abstract Background: The mitogen-activated protein kinase (MAPK) cascade plays an important role in plant development and stress response. In particular, MAPK kinases (MAPKKs/MKKs) have been reported to play a crucial role in MAPK cascades that mediate a variety of stress responses in plants. Few MKKs in grapevine (Vitis vinifera), however, have been functionally characterized.Results: In the present study, five MKK members in grapevine (‘Pinot Noir’) were identified, cloned and designated as VvMKK1-VvMKK5. A phylogenetic analysis grouped these into four sub-families based on the similarity of their conserved motifs and gene structure to other Arabidopsis MKK members. RT-qPCR results indicated that the expression of VvMKK1, VvMKK2, VvMKK4, and VvMKK5 was up-regulated in mature leaves, young leaves, and roots, but exhibited low expression levels in leaf petioles. VvMKK2, VvMKK3, and VvMKK5 genes were differentially up-regulated when grapevine leaves were inoculated with spores of Erisyphe necator, or treated with SA, ETH, H2O2, or exposed to drought, indicating that these genes may be involved in a variety of signaling pathways. Over expression of VvMKK2 and VvMKK4 genes in transgenic Arabidopsis plants resulted in the production of seeds with a significantly higher germination and survival rate, and better seedling growth under stress conditions than wild-type plants. Overexpression of VvMKK2 in Arabidopsis resulted in improved salt and drought stress tolerance while overexpression of VvMKK4 improved salt stress tolerance.Conclusions: Results of the present investigation provide a better understanding of the interaction and function of MAPKKK-MAPKK-MAPK genes at the transcriptional level in grapevine and led to the identification of candidate genes for improved drought and salt stress in grapes.

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Melatonin-Mediated Regulation of Growth and Antioxidant Capacity in Salt-Tolerant Naked Oat under Salt Stress

International Journal of Molecular Sciences ◽

10.3390/ijms20051176 ◽

2019 ◽

Vol 20 (5) ◽

pp. 1176 ◽

Cited By ~ 14

Author(s):

Wenying Gao ◽

Zheng Feng ◽

Qingqing Bai ◽

Jinjin He ◽

Yingjuan Wang

Keyword(s):

Salt Stress ◽

Antioxidant Capacity ◽

Stress Responses ◽

Nacl Stress ◽

Mitogen Activated Protein Kinase ◽

Naked Oat ◽

Exogenous Application ◽

Transcription Factor Genes ◽

Mitogen Activated Protein

Melatonin (MT; N-acetyl-5-methoxytryptamine) is a pleiotropic signaling molecule that has been demonstrated to play an important role in plant growth, development, and regulation of environmental stress responses. Studies have been conducted on the role of the exogenous application of MT in a few species, but the potential mechanisms of MT-mediated stress tolerance under salt stress are still largely unknown. In this study, naked oat seedlings under salt stress (150 mM NaCl) were pretreated with two different concentrations of MT (50 and 100 μM), and the effects of MT on the growth and antioxidant capacity of naked oat seedlings were analyzed to explore the regulatory effect of MT on salt tolerance. The results showed that pretreating with different concentrations of MT promoted the growth of seedlings in response to 150 mM NaCl. Different concentrations of MT reduced hydrogen peroxide, superoxide anion, and malondialdehyde contents. The exogenous application of MT also increased superoxide dismutase, peroxidase, catalase, and ascorbate peroxide activities. Chlorophyll content, leaf area, leaf volume, and proline increased in the leaves of naked oat seedlings under 150 mM NaCl stress. MT upregulated the expression levels of the lipid peroxidase genes lipoxygenase and peroxygenase, a chlorophyll biosynthase gene (ChlG), the mitogen-activated protein kinase genes Asmap1 and Aspk11, and the transcription factor genes (except DREB2), NAC, WRKY1, WRKY3, and MYB in salt-exposed MT-pretreated seedlings when compared with seedlings exposed to salt stress alone. These results demonstrate an important role of MT in the relief of salt stress and, therefore, provide a reference for managing salinity in naked oat.

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Identification of a novel mitogen-activated protein kinase kinase gene (MKK2) in the oilseed rape Brassica campestris

Biologia ◽

10.2478/s11756-014-0455-8 ◽

2014 ◽

Vol 69 (11) ◽

Author(s):

Teng Zhang ◽

Yuan Wang ◽

Juan Wang ◽

Xiao Xia ◽

Ning Yang ◽

...

Keyword(s):

Protein Kinase ◽

Stress Responses ◽

Brassica Campestris ◽

Plant Stress ◽

Mitogen Activated Protein Kinase ◽

Biotic And Abiotic Stress ◽

Kinase Gene ◽

Isolation And Characterization ◽

Mapk Cascades ◽

Mitogen Activated Protein

AbstractMitogen-activated protein kinase (MAPK) cascades participate in various processes, including plant growth and development as well as biotic and abiotic stress responses. MAPK kinases (MKKs), which link MPKs and MPKK kinases, are involved in MAPK cascades by mediating various plant stress responses. However, only a few MKKs from Brassica campestris (rape) have been functionally characterized. This study delivers the results from isolation and characterization of a novel gene, MKK2, from rape. Bioinformatics analysis revealed that the cDNA length of MKK2 is 1,344 bp with an open reading frame of 1,068 bp, which encodes a polypeptide containing 355 amino acids. The obtained MKK2 exhibited a predicted molecular mass of 39.3 kDa and an isoelectric point of 6.8. Quantitative real-time polymerase chain reaction analysis revealed that MKK2 expression can be induced by cold and salt. Western blot analysis revealed that MKK2 protein expression can be induced by cold, salt, and UV-B radiation. The MKK2 protein was localized in the nucleus. These results suggest that MKK2 is important for the regulation of cold- and salt-stress responses in plants.

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3pK, a novel mitogen-activated protein (MAP) kinase-activated protein kinase, is targeted by three MAP kinase pathways.

Molecular and Cellular Biology ◽

10.1128/mcb.16.12.6687 ◽

1996 ◽

Vol 16 (12) ◽

pp. 6687-6697 ◽

Cited By ~ 128

Author(s):

S Ludwig ◽

K Engel ◽

A Hoffmeyer ◽

G Sithanandam ◽

B Neufeld ◽

...

Keyword(s):

Protein Kinase ◽

Map Kinase ◽

Stress Responses ◽

Specific Inhibitor ◽

Mitogen Activated Protein Kinase ◽

Mapk Cascades ◽

Mitogen Activated Protein ◽

Stimulation Of

Recently we have identified a mitogen-activated protein kinase (MAPK)-activated protein kinase, named 3pK (G. Sithanandam, F. Latif, U. Smola, R. A. Bernal, F.-M. Duh, H. Li, I. Kuzmin, V. Wixler, L. Geil, S. Shresta, P. A. Lloyd, S. Bader, Y. Sekido, K. D. Tartof, V. I. Kashuba, E. R. Zabarovsky, M. Dean, G. Klein, B. Zbar, M. I. Lerman, J. D. Minna, U. R. Rapp, and A. Allikmets, Mol. Cell. Biol. 16:868-876, 1996). In vitro characterization of the kinase revealed that 3pK is activated by ERK. It was further shown that 3pK is phosphorylated in vivo after stimulation of cells with serum. However, the in vivo relevance of this observation in terms of involvement of the Raf/MEK/ERK cascade has not been established. Here we show that 3pK is activated in vivo by the growth inducers serum and tetradecanoyl phorbol acetate in promyelocytic HL60 cells and transiently transfected embryonic kidney 293 cells. Activation of 3pK was Raf dependent and was mediated by the Raf/MEK/ERK kinase cascade. 3pK was also shown to be activated after stress stimulation of cells. In vitro studies with recombinant proteins demonstrate that in addition to ERK, members of other subgroups of the MAPK family, namely, p38RK and Jun-N-terminal kinases/stress-activated protein kinases, were also able to phosphorylate and activate 3pK. Cotransfection experiments as well as the use of a specific inhibitor of p38RK showed that these in vitro upstream activators also function in vivo, identifying 3pK as the first kinase to be activated through all three MAPK cascades. Thus, 3pK is a novel convergence point of different MAPK pathways and could function as an integrative element of signaling in both mitogen and stress responses.

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Plant Mitogen-Activated Protein Kinase Cascades in Environmental Stresses

International Journal of Molecular Sciences ◽

10.3390/ijms22041543 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1543

Author(s):

Li Lin ◽

Jian Wu ◽

Mingyi Jiang ◽

Youping Wang

Keyword(s):

Protein Kinase ◽

Environmental Stress ◽

Stress Responses ◽

Extreme Temperature ◽

Environmental Stresses ◽

Mitogen Activated Protein Kinase ◽

Hormone Production ◽

Mapk Cascades ◽

Stress Signal ◽

Mitogen Activated Protein

Due to global warming and population growth, plants need to rescue themselves, especially in unfavorable environments, to fulfill food requirements because they are sessile organisms. Stress signal sensing is a crucial step that determines the appropriate response which, ultimately, determines the survival of plants. As important signaling modules in eukaryotes, plant mitogen-activated protein kinase (MAPK) cascades play a key role in regulating responses to the following four major environmental stresses: high salinity, drought, extreme temperature and insect and pathogen infections. MAPK cascades are involved in responses to these environmental stresses by regulating the expression of related genes, plant hormone production and crosstalk with other environmental stresses. In this review, we describe recent major studies investigating MAPK-mediated environmental stress responses. We also highlight the diverse function of MAPK cascades in environmental stress. These findings help us understand the regulatory network of MAPKs under environmental stress and provide another strategy to improve stress resistance in crops to ensure food security.

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Update on the Roles of Rice MAPK Cascades

International Journal of Molecular Sciences ◽

10.3390/ijms22041679 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1679

Author(s):

Jie Chen ◽

Lihan Wang ◽

Meng Yuan

Keyword(s):

Signal Transduction ◽

Abiotic Stress ◽

Stress Responses ◽

Molecular Mechanisms ◽

Developmental Regulation ◽

Mitogen Activated Protein Kinase ◽

Biotic And Abiotic Stress ◽

Mapk Kinase ◽

Mapk Cascades ◽

Mitogen Activated Protein

The mitogen-activated protein kinase (MAPK) cascades have been validated playing critical roles in diverse aspects of plant biology, from growth and developmental regulation, biotic and abiotic stress responses, to phytohormone signal transduction or responses. A classical MAPK cascade consists of a MAPK kinase kinase (MAPKKK), a MAPK kinase (MAPKK), and a MAPK. From the 75 MAPKKKs, eight MAPKKs, and 15 MAPKs of rice, a number of them have been functionally deciphered. Here, we update recent advances in knowledge of the roles of rice MAPK cascades, including their components and complicated action modes, their diversified functions controlling rice growth and developmental responses, coordinating resistance to biotic and abiotic stress, and conducting phytohormone signal transduction. Moreover, we summarize several complete MAPK cascades that harbor OsMAPKKK-OsMAPKK-OsMAPK, their interaction with different upstream components and their phosphorylation of diverse downstream substrates to fulfill their multiple roles. Furthermore, we state a comparison of networks of rice MAPK cascades from signal transduction crosstalk to the precise selection of downstream substrates. Additionally, we discuss putative concerns for elucidating the underlying molecular mechanisms and molecular functions of rice MAPK cascades in the future.

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Cloning, molecular and functional characterization by overexpression in Arabidopsis of MAPKK genes from grapevine (Vitis vinifera)

10.21203/rs.2.14827/v4 ◽

2020 ◽

Author(s):

Gang Wang ◽

Ying-hai Liang ◽

Ji-yu Zhang ◽

Zong-Ming (Max) Cheng

Keyword(s):

Salt Stress ◽

Stress Tolerance ◽

Functional Characterization ◽

Pinot Noir ◽

Transcriptional Level ◽

Environmental Signals ◽

Grapevine Cultivar ◽

Drought And Salt Stress ◽

Mitogen Activated Protein ◽

And Function

Abstract Background : The mitogen-activated protein kinases (MAPKs), as a part of the MAPKKK-MAPKK-MAPK cascade, play crucial roles in plant development as an intracellular signal transduction pathways to respond various environmental signals. However, few MAPKK have been functionally characterized in grapevine. Results : In the study, five MAPKK (MKK) members were identified in grapevine (cultivar ‘Pinot Noir’), cloned and designated as VvMKK1-VvMKK5. A phylogenetic analysis grouped them into four sub-families based on the similarity of their conserved motifs and gene structure to Arabidopsis MAPKK members. qRT-PCR results indicated that the expression of VvMKK1, VvMKK2, VvMKK4, and VvMKK5 were up-regulated in mature leaf and young blades, and roots, but exhibited low expression in leaf petioles. VvMKK2, VvMKK3, and VvMKK5 genes were differentially up-regulated when grapevine leaves were inoculated with spores of Erisyphe necator , or treated with salicylic acid (SA), ethylene (ETH), H 2 O 2, or exposed to drought, indicating that these genes may be involved in a variety of signaling pathways. Over expression of VvMKK2 and VvMKK4 genes in transgenic Arabidopsis plants resulted in the production of seeds with a significantly higher germination and survival rate, and better seedling growth under stress conditions than wild-type plants. Overexpression of VvMKK2 in Arabidopsis improved salt and drought stress tolerance while overexpression of VvMKK4 only improved salt stress tolerance. Conclusions: Results of the present investigation provide a better understanding of the interaction and function of MAPKKK-MAPKK-MAPK genes at the transcriptional level in grapevine and led to the identification of candidate genes for drought and salt stress in grapes.

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Cloning, molecular and functional characterization by overexpression in Arabidopsis of MAPKK genes from grapevine (Vitis vinifera)

10.21203/rs.2.14827/v3 ◽

2020 ◽

Author(s):

Gang Wang ◽

Ying-hai Liang ◽

Ji-yu Zhang ◽

Zong-Ming (Max) Cheng

Keyword(s):

Salt Stress ◽

Stress Tolerance ◽

Functional Characterization ◽

Pinot Noir ◽

Transcriptional Level ◽

Environmental Signals ◽

Grapevine Cultivar ◽

Drought And Salt Stress ◽

Mitogen Activated Protein ◽

And Function

Abstract Background: The mitogen-activated protein kinases (MAPKs), as a part of the MAPKKK-MAPKK-MAPK cascade, play crucial roles in plant development as an intracellular signal transduction pathways to respond various environmental signals. However, few MAPKK have been functionally characterized in grapevine.Results: In the study, five MAPKK (MKK) members were identified in grapevine (cultivar ‘Pinot Noir’), cloned and designated as VvMKK1-VvMKK5. A phylogenetic analysis grouped them into four sub-families based on the similarity of their conserved motifs and gene structure to Arabidopsis MAPKK members. qRT-PCR results indicated that the expression of VvMKK1, VvMKK2, VvMKK4, and VvMKK5 were up-regulated in mature leaf and young blades, and roots, but exhibited low expression in leaf petioles. VvMKK2, VvMKK3, and VvMKK5 genes were differentially up-regulated when grapevine leaves were inoculated with spores of Erisyphe necator, or treated with salicylic acid (SA), ethylene (ETH), H2O2, or exposed to drought, indicating that these genes may be involved in a variety of signaling pathways. Over expression of VvMKK2 and VvMKK4 genes in transgenic Arabidopsis plants resulted in the production of seeds with a significantly higher germination and survival rate, and better seedling growth under stress conditions than wild-type plants. Overexpression of VvMKK2 in Arabidopsis improved salt and drought stress tolerance while overexpression of VvMKK4 only improved salt stress tolerance.Conclusions: Results of the present investigation provide a better understanding of the interaction and function of MAPKKK-MAPKK-MAPK genes at the transcriptional level in grapevine and led to the identification of candidate genes for drought and salt stress in grapes.

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Chlorosis seedling lethality 1 encoding a MAP3K protein is essential for chloroplast development in rice

BMC Plant Biology ◽

10.1186/s12870-021-03404-9 ◽

2022 ◽

Vol 22 (1) ◽

Author(s):

Jiayan Liang ◽

Qiuxin Zhang ◽

Yiran Liu ◽

Jingjing Zhang ◽

Wenyi Wang ◽

...

Keyword(s):

Stress Responses ◽

Chloroplast Development ◽

Direct Interaction ◽

Mitogen Activated Protein Kinase ◽

Starch Granules ◽

Altered Expression ◽

Mapk Cascades ◽

Mitogen Activated Protein ◽

Seedling Lethality ◽

Eukaryotic Organisms

Abstract Background Mitogen-activated protein kinase (MAPK) cascades are conserved signaling modules in eukaryotic organisms and play essential roles in immunity and stress responses. However, the role of MAPKs in chloroplast development remains to be evidently established. Results In this study, a rice chlorosis seedling lethality 1 (csl1) mutant with a Zhonghua11 (ZH11, japonica) background was isolated. Seedlings of the mutant were characterized by chlorotic leaves and death after the trefoil stage, and chloroplasts were observed to contain accumulated starch granules. Molecular cloning revealed that OsCSL1 encoded a MAPK kinase kinase22 (MKKK22) targeted to the endoplasmic reticulum (ER), and functional complementation of OsCSL1 was found to restore the normal phenotype in csl1 plants. The CRISPR/Cas9 technology was used for targeted disruption of OsCSL1, and the OsCSL1-Cas9 lines obtained therein exhibited yellow seedlings which phenocopied the csl1 mutant. CSL1/MKKK22 was observed to establish direct interaction with MKK4, and altered expression of MKK1 and MKK4 was detected in the csl1 mutant. Additionally, disruption of OsCSL1 led to reduced expression of chloroplast-associated genes, including chlorophyll biosynthetic genes, plastid-encoded RNA polymerases, nuclear-encoded RNA polymerase, and nuclear-encoded chloroplast genes. Conclusions The findings of this study revealed that OsCSL1 played roles in regulating the expression of multiple chloroplast synthesis-related genes, thereby affecting their functions, and leading to wide-ranging defects, including chlorotic seedlings and severely disrupted chloroplasts containing accumulated starch granules.

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Dual control of MAPK activities by AP2C1 and MKP1 MAPK phosphatases regulates defence responses in Arabidopsis

10.1101/2021.06.03.446911 ◽

2021 ◽

Author(s):

Zahra Ayatollahi ◽

Vaiva Kazanaviciute ◽

Volodymyr Shubchynskyy ◽

Kotryna Kvederaviciute ◽

Manfred Schwanninger ◽

...

Keyword(s):

Stress Responses ◽

Stress Hormones ◽

Plant Stress ◽

Mitogen Activated Protein Kinase ◽

Environmental Signals ◽

Mapk Cascades ◽

Mitogen Activated Protein ◽

Plant Stress Responses ◽

Mapk Phosphatases ◽

Defence Responses

Mitogen-activated protein kinase (MAPK) cascades transmit environmental signals and induce stress and defence responses in plants. These signalling cascades are negatively controlled by specific phosphatases of the type 2C Ser/Thr protein phosphatase (PP2C) and dual-specificity phosphatase (DSP) families that inactivate stress-induced MAPKs; however, the interplay between phosphatases of these different types has remained unknown. Our work reveals that different Arabidopsis MAPK phosphatases, the PP2C-type AP2C1 and the DSP-type MKP1, exhibit both specific and overlapping functions in plant stress responses. Each single mutant and ap2c1 mkp1 double mutant displayed enhanced wound-induced activation of MAPKs MPK3, MPK4, and MPK6, as well as induction of a set of transcription factors. Moreover, ap2c1 mkp1 double mutants show an autoimmune-like response, associated with elevated levels the stress hormones salicylic acid and ethylene, and of the phytoalexin camalexin. Interestingly, this phenotype is reduced in ap2c1 mkp1 mpk6 triple mutants, suggesting that the autoimmune-like response is due to MPK6 misregulation. We conclude that the evolutionarily distant MAPK phosphatases AP2C1 and MKP1 contribute crucially to the tight control of MPK6 activity, ensuring appropriately balanced stress signalling and suppression of autoimmune-like responses during plant growth and development.

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Cloning, molecular and functional characterization by overexpression in Arabidopsis of MAPKK genes from grapevine (Vitis vinifera)

10.21203/rs.2.14827/v2 ◽

2020 ◽

Author(s):

Gang Wang ◽

Ying-hai Liang ◽

Ji-yu Zhang ◽

Zong-Ming (Max) Cheng

Keyword(s):

Salt Stress ◽

Stress Tolerance ◽

Functional Characterization ◽

Pinot Noir ◽

Transcriptional Level ◽

Environmental Signals ◽

Grapevine Cultivar ◽

Drought And Salt Stress ◽

Mitogen Activated Protein ◽

And Function

Abstract Background : The mitogen-activated protein kinases (MAPKs), as a part of the MAPKKK-MAPKK-MAPK cascade play crucial roles in plant development as an intracellular signal transduction pathways to respond various environmental signals. However, few MAPKK have been functionally characterized in grapevine. Results : In the study, five MAPKK (MKK) members were identified in grapevine (cultivar ‘Pinot Noir’), cloned and designated as VvMKK1-VvMKK5. A phylogenetic analysis grouped them into four sub-families based on the similarity of their conserved motifs and gene structure to Arabidopsis MAPKK members. qRT-PCR results indicated that the expression of VvMKK1, VvMKK2, VvMKK4, and VvMKK5 were up-regulated in mature leaf and young blades, and roots, but exhibited low expression in leaf petioles. VvMKK2, VvMKK3, and VvMKK5 genes were differentially up-regulated when grapevine leaves were inoculated with spores of Erisyphe necator , or treated with salicylic acid (SA), ethylene (ETH), H 2 O 2, or exposed to drought, indicating that these genes may be involved in a variety of signaling pathways. Over expression of VvMKK2 and VvMKK4 genes in transgenic Arabidopsis plants resulted in the production of seeds with a significantly higher germination and survival rate, and better seedling growth under stress conditions than wild-type plants. Overexpression of VvMKK2 in Arabidopsis improved salt and drought stress tolerance while overexpression of VvMKK4 only improved salt stress tolerance. Conclusions: Results of the present investigation provide a better understanding of the interaction and function of MAPKKK-MAPKK-MAPK genes at the transcriptional level in grapevine and led to the identification of candidate genes for drought and salt stress in grapes.

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