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GhMPK16, a novel stress-responsive group D MAPK gene from cotton, is involved in disease resistance and drought sensitivity.

Shi J, Zhang L, An H, Wu C, Guo X - BMC Mol. Biol. (2011)

Bottom Line: In this study, we isolated and characterised GhMPK16, the first group D MAPK gene found in cotton.Furthermore, transgenic Arabidopsis showed reduced drought tolerance and rapid H2O2 accumulation.These results suggest that GhMPK16 might be involved in multiple signal transduction pathways, including biotic and abiotic stress signaling pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, Shandong Agricultural University, Taian, Shandong 271018, China.

ABSTRACT

Background: Mitogen-activated protein kinase (MAPK) cascades play pivotal roles in mediating biotic and abiotic stress responses. In plants, MAPKs are classified into four major groups (A-D) according to their sequence homology and conserved phosphorylation motifs. Members of group A and B have been extensively characterized, but little information on the group D MAPKs has been reported.

Results: In this study, we isolated and characterised GhMPK16, the first group D MAPK gene found in cotton. Southern blot analysis suggests GhMPK16 is single copy in the cotton genome, and RNA blot analysis indicates that GhMPK16 transcripts accumulate following pathogen infection and treatment with multiple defense-related signal molecules. The analysis of the promoter region of GhMPK16 revealed a group of putative cis-acting elements related to stress responses. Subcellular localization analysis suggests that GhMPK16 acts in the nucleus. Transgenic Arabidopsis overexpressing GhMPK16 displayed significant resistance to fungi (Colletotrichum nicotianae and Alternaria alternata) and bacteria (Pseudomonas solanacearum) pathogen, and the transcripts of pathogen-related (PR) genes were more rapidly and strongly induced in the transgenic plants. Furthermore, transgenic Arabidopsis showed reduced drought tolerance and rapid H2O2 accumulation.

Conclusion: These results suggest that GhMPK16 might be involved in multiple signal transduction pathways, including biotic and abiotic stress signaling pathways.

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Related in: MedlinePlus

Subcellular localization of the GhMPK16 protein in onion epidermal cells. (A) Schematic diagram of the 35S-GhMPK16::GFP and 35S-GFP fusion construct. (B) Transient expression of 35S-GFP and 35S-GhMPK16::GFP in onion epidermal cells. Cells were analyzed by laser confocal microscopy 12 h after particle bombardment. The nuclei of the onion cells were visualised by DAPI staining. Bar = 20 μm.
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Figure 3: Subcellular localization of the GhMPK16 protein in onion epidermal cells. (A) Schematic diagram of the 35S-GhMPK16::GFP and 35S-GFP fusion construct. (B) Transient expression of 35S-GFP and 35S-GhMPK16::GFP in onion epidermal cells. Cells were analyzed by laser confocal microscopy 12 h after particle bombardment. The nuclei of the onion cells were visualised by DAPI staining. Bar = 20 μm.

Mentions: To reveal the cellular localization of GhMPK16, a reporter gene encoding GFP was fused to GhMPK16 and placed under the control of the CaMV35S promoter, and immunoblot analysis indicated that GhMPK16::GFP was an integrated fusion protein (Additional file 2: supplementary Figure S2). The biolistic transformation system was used for a transient assay in onion epidermal cells. The nuclear localization of GFP-conjugated GhMPK16 was confirmed in individual transgenic cells by GFP fluorescence, using DAPI staining to detect the nuclei and interference contrast images to detect whole-cell structures. As shown in Figure 3B, the 35S-GhMPK16::GFP construct localized to the nucleus, and the 35S-GFP control construct showed GFP signals in both the cytoplasm and the nucleus. In addition, a program that predicts the subcellular localization of proteins (http://www.bioinfo.tsinghua.edu.cn/SubLoc/) predicted that GhMPK16 is localized in the nucleus with an expected accuracy of nearly 74%. These results indicate that the GhMPK16 protein is likely localized in the nucleus.


GhMPK16, a novel stress-responsive group D MAPK gene from cotton, is involved in disease resistance and drought sensitivity.

Shi J, Zhang L, An H, Wu C, Guo X - BMC Mol. Biol. (2011)

Subcellular localization of the GhMPK16 protein in onion epidermal cells. (A) Schematic diagram of the 35S-GhMPK16::GFP and 35S-GFP fusion construct. (B) Transient expression of 35S-GFP and 35S-GhMPK16::GFP in onion epidermal cells. Cells were analyzed by laser confocal microscopy 12 h after particle bombardment. The nuclei of the onion cells were visualised by DAPI staining. Bar = 20 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3117701&req=5

Figure 3: Subcellular localization of the GhMPK16 protein in onion epidermal cells. (A) Schematic diagram of the 35S-GhMPK16::GFP and 35S-GFP fusion construct. (B) Transient expression of 35S-GFP and 35S-GhMPK16::GFP in onion epidermal cells. Cells were analyzed by laser confocal microscopy 12 h after particle bombardment. The nuclei of the onion cells were visualised by DAPI staining. Bar = 20 μm.
Mentions: To reveal the cellular localization of GhMPK16, a reporter gene encoding GFP was fused to GhMPK16 and placed under the control of the CaMV35S promoter, and immunoblot analysis indicated that GhMPK16::GFP was an integrated fusion protein (Additional file 2: supplementary Figure S2). The biolistic transformation system was used for a transient assay in onion epidermal cells. The nuclear localization of GFP-conjugated GhMPK16 was confirmed in individual transgenic cells by GFP fluorescence, using DAPI staining to detect the nuclei and interference contrast images to detect whole-cell structures. As shown in Figure 3B, the 35S-GhMPK16::GFP construct localized to the nucleus, and the 35S-GFP control construct showed GFP signals in both the cytoplasm and the nucleus. In addition, a program that predicts the subcellular localization of proteins (http://www.bioinfo.tsinghua.edu.cn/SubLoc/) predicted that GhMPK16 is localized in the nucleus with an expected accuracy of nearly 74%. These results indicate that the GhMPK16 protein is likely localized in the nucleus.

Bottom Line: In this study, we isolated and characterised GhMPK16, the first group D MAPK gene found in cotton.Furthermore, transgenic Arabidopsis showed reduced drought tolerance and rapid H2O2 accumulation.These results suggest that GhMPK16 might be involved in multiple signal transduction pathways, including biotic and abiotic stress signaling pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, Shandong Agricultural University, Taian, Shandong 271018, China.

ABSTRACT

Background: Mitogen-activated protein kinase (MAPK) cascades play pivotal roles in mediating biotic and abiotic stress responses. In plants, MAPKs are classified into four major groups (A-D) according to their sequence homology and conserved phosphorylation motifs. Members of group A and B have been extensively characterized, but little information on the group D MAPKs has been reported.

Results: In this study, we isolated and characterised GhMPK16, the first group D MAPK gene found in cotton. Southern blot analysis suggests GhMPK16 is single copy in the cotton genome, and RNA blot analysis indicates that GhMPK16 transcripts accumulate following pathogen infection and treatment with multiple defense-related signal molecules. The analysis of the promoter region of GhMPK16 revealed a group of putative cis-acting elements related to stress responses. Subcellular localization analysis suggests that GhMPK16 acts in the nucleus. Transgenic Arabidopsis overexpressing GhMPK16 displayed significant resistance to fungi (Colletotrichum nicotianae and Alternaria alternata) and bacteria (Pseudomonas solanacearum) pathogen, and the transcripts of pathogen-related (PR) genes were more rapidly and strongly induced in the transgenic plants. Furthermore, transgenic Arabidopsis showed reduced drought tolerance and rapid H2O2 accumulation.

Conclusion: These results suggest that GhMPK16 might be involved in multiple signal transduction pathways, including biotic and abiotic stress signaling pathways.

Show MeSH
Related in: MedlinePlus