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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

Sequence analysis of GhMPK16. (A) Alignment of the deduced GhMPK16 protein sequence with other known plant group D MAPKs. Amino acid sequences used in the analysis are from Arabidopsis thaliana (AtMPK16, NP_197402), Triticum aestivum (TaMAPK2, ABC54585), Zea mays (ZmMPK6, NP_001105238), Oryza sativa (OsMPK16-1, ACF06191), Gossypium hirsutum (GhMPK16, FJ966888). These sequences were aligned using the DNAman 6.0 program. Identical and similar amino acids were shaded in black and gray, respectively. The protein kinase subdomains are indicated by Roman numerals (I to XI) over the sequences. The activation-loop is boxed, and the phosphorylation motif (TDY) is marked by an arrow. (B) The phylogenetic relationships of GhMPK16 and other plant MAPKs. Numbers above or below the branches indicate bootstrap values (>50%) from 1,000 replicates. The amino acid sequences of MAPKs used for construction of the tree are deposited to the GenBank database under the following accession numbers: AtMPK3 (NP_190150); NtWIPK (BAA09600); PsMAPK3 (AAF73236); OsMPK5 (AAL87689); AtMPK6 (NP_181907); NtSIPK (AAB58396); NtNTF6 (CAA58760); AtMPK5 (NP_567378); AtMPK4 (NP_192046); OsMPK6 (NP_922504); AtMPK7 (NP_179409); OsMPK3 (ABH01189); NtNTF3 (CAA49592); AtMPK1 (NP_172492); GhMAPK (ABA00652); AtMPK18 (NP_175756); OsMPK8 (CAD54742); AtMPK17 (NP_001030941); AtMPK15 (NP_565070); AtMPK9 (NP_566595); OsMPK12 (AAD52659); AtMPK16 (NP_197402); GhMPK16 (FJ966889); OsMPK15 (ACD76441); ZmMPK6 (NP_001105238); OsMPK16-1 (ACF06191); TaMAPK2 (ABC54585). At, Arabidopsis thaliana; Nt, Nicotiana tobacum; Ps, Pisum sativum; Ta, Triticum aestivum; Zm, Zea mays; Os, Oryza sativa; Gh, Gossypium hirsutum. The Ser/Thr kinase domain is indicated as gray and contains part of the activation-loop motif; the CD domain required for MAPK docking is indicated in black. GhMPK16 is boxed in the diagram and classified as a member of the TDY group.
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Figure 1: Sequence analysis of GhMPK16. (A) Alignment of the deduced GhMPK16 protein sequence with other known plant group D MAPKs. Amino acid sequences used in the analysis are from Arabidopsis thaliana (AtMPK16, NP_197402), Triticum aestivum (TaMAPK2, ABC54585), Zea mays (ZmMPK6, NP_001105238), Oryza sativa (OsMPK16-1, ACF06191), Gossypium hirsutum (GhMPK16, FJ966888). These sequences were aligned using the DNAman 6.0 program. Identical and similar amino acids were shaded in black and gray, respectively. The protein kinase subdomains are indicated by Roman numerals (I to XI) over the sequences. The activation-loop is boxed, and the phosphorylation motif (TDY) is marked by an arrow. (B) The phylogenetic relationships of GhMPK16 and other plant MAPKs. Numbers above or below the branches indicate bootstrap values (>50%) from 1,000 replicates. The amino acid sequences of MAPKs used for construction of the tree are deposited to the GenBank database under the following accession numbers: AtMPK3 (NP_190150); NtWIPK (BAA09600); PsMAPK3 (AAF73236); OsMPK5 (AAL87689); AtMPK6 (NP_181907); NtSIPK (AAB58396); NtNTF6 (CAA58760); AtMPK5 (NP_567378); AtMPK4 (NP_192046); OsMPK6 (NP_922504); AtMPK7 (NP_179409); OsMPK3 (ABH01189); NtNTF3 (CAA49592); AtMPK1 (NP_172492); GhMAPK (ABA00652); AtMPK18 (NP_175756); OsMPK8 (CAD54742); AtMPK17 (NP_001030941); AtMPK15 (NP_565070); AtMPK9 (NP_566595); OsMPK12 (AAD52659); AtMPK16 (NP_197402); GhMPK16 (FJ966889); OsMPK15 (ACD76441); ZmMPK6 (NP_001105238); OsMPK16-1 (ACF06191); TaMAPK2 (ABC54585). At, Arabidopsis thaliana; Nt, Nicotiana tobacum; Ps, Pisum sativum; Ta, Triticum aestivum; Zm, Zea mays; Os, Oryza sativa; Gh, Gossypium hirsutum. The Ser/Thr kinase domain is indicated as gray and contains part of the activation-loop motif; the CD domain required for MAPK docking is indicated in black. GhMPK16 is boxed in the diagram and classified as a member of the TDY group.

Mentions: A BLAST search (http://www.ncbi.nlm.nih.gov/BLAST) and multi-alignment analysis revealed that GhMPK16 is highly related to other group D MAPKs, sharing a homology of 81.48% to AtMPK16, 72.18% to TaMAPK2, 74.56% to OsMPK16-1 and 76.02% to ZmMPK6 (Figure 1A). Moreover, as with other known plant group D MAPKs, GhMPK16 has the same family signature of 11 conserved subdomains, an activation loop (T-loop), a phosphorylation motif (TDY motif) in the T-loop, and an extended C-terminal region relative to groups A, B and C. Additionally, GhMPK16 also lacks a C-terminal CD domain, a feature conserved in MAPKs belonging to the other groups (A, B, C) [4].


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)

Sequence analysis of GhMPK16. (A) Alignment of the deduced GhMPK16 protein sequence with other known plant group D MAPKs. Amino acid sequences used in the analysis are from Arabidopsis thaliana (AtMPK16, NP_197402), Triticum aestivum (TaMAPK2, ABC54585), Zea mays (ZmMPK6, NP_001105238), Oryza sativa (OsMPK16-1, ACF06191), Gossypium hirsutum (GhMPK16, FJ966888). These sequences were aligned using the DNAman 6.0 program. Identical and similar amino acids were shaded in black and gray, respectively. The protein kinase subdomains are indicated by Roman numerals (I to XI) over the sequences. The activation-loop is boxed, and the phosphorylation motif (TDY) is marked by an arrow. (B) The phylogenetic relationships of GhMPK16 and other plant MAPKs. Numbers above or below the branches indicate bootstrap values (>50%) from 1,000 replicates. The amino acid sequences of MAPKs used for construction of the tree are deposited to the GenBank database under the following accession numbers: AtMPK3 (NP_190150); NtWIPK (BAA09600); PsMAPK3 (AAF73236); OsMPK5 (AAL87689); AtMPK6 (NP_181907); NtSIPK (AAB58396); NtNTF6 (CAA58760); AtMPK5 (NP_567378); AtMPK4 (NP_192046); OsMPK6 (NP_922504); AtMPK7 (NP_179409); OsMPK3 (ABH01189); NtNTF3 (CAA49592); AtMPK1 (NP_172492); GhMAPK (ABA00652); AtMPK18 (NP_175756); OsMPK8 (CAD54742); AtMPK17 (NP_001030941); AtMPK15 (NP_565070); AtMPK9 (NP_566595); OsMPK12 (AAD52659); AtMPK16 (NP_197402); GhMPK16 (FJ966889); OsMPK15 (ACD76441); ZmMPK6 (NP_001105238); OsMPK16-1 (ACF06191); TaMAPK2 (ABC54585). At, Arabidopsis thaliana; Nt, Nicotiana tobacum; Ps, Pisum sativum; Ta, Triticum aestivum; Zm, Zea mays; Os, Oryza sativa; Gh, Gossypium hirsutum. The Ser/Thr kinase domain is indicated as gray and contains part of the activation-loop motif; the CD domain required for MAPK docking is indicated in black. GhMPK16 is boxed in the diagram and classified as a member of the TDY group.
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Related In: Results  -  Collection

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Figure 1: Sequence analysis of GhMPK16. (A) Alignment of the deduced GhMPK16 protein sequence with other known plant group D MAPKs. Amino acid sequences used in the analysis are from Arabidopsis thaliana (AtMPK16, NP_197402), Triticum aestivum (TaMAPK2, ABC54585), Zea mays (ZmMPK6, NP_001105238), Oryza sativa (OsMPK16-1, ACF06191), Gossypium hirsutum (GhMPK16, FJ966888). These sequences were aligned using the DNAman 6.0 program. Identical and similar amino acids were shaded in black and gray, respectively. The protein kinase subdomains are indicated by Roman numerals (I to XI) over the sequences. The activation-loop is boxed, and the phosphorylation motif (TDY) is marked by an arrow. (B) The phylogenetic relationships of GhMPK16 and other plant MAPKs. Numbers above or below the branches indicate bootstrap values (>50%) from 1,000 replicates. The amino acid sequences of MAPKs used for construction of the tree are deposited to the GenBank database under the following accession numbers: AtMPK3 (NP_190150); NtWIPK (BAA09600); PsMAPK3 (AAF73236); OsMPK5 (AAL87689); AtMPK6 (NP_181907); NtSIPK (AAB58396); NtNTF6 (CAA58760); AtMPK5 (NP_567378); AtMPK4 (NP_192046); OsMPK6 (NP_922504); AtMPK7 (NP_179409); OsMPK3 (ABH01189); NtNTF3 (CAA49592); AtMPK1 (NP_172492); GhMAPK (ABA00652); AtMPK18 (NP_175756); OsMPK8 (CAD54742); AtMPK17 (NP_001030941); AtMPK15 (NP_565070); AtMPK9 (NP_566595); OsMPK12 (AAD52659); AtMPK16 (NP_197402); GhMPK16 (FJ966889); OsMPK15 (ACD76441); ZmMPK6 (NP_001105238); OsMPK16-1 (ACF06191); TaMAPK2 (ABC54585). At, Arabidopsis thaliana; Nt, Nicotiana tobacum; Ps, Pisum sativum; Ta, Triticum aestivum; Zm, Zea mays; Os, Oryza sativa; Gh, Gossypium hirsutum. The Ser/Thr kinase domain is indicated as gray and contains part of the activation-loop motif; the CD domain required for MAPK docking is indicated in black. GhMPK16 is boxed in the diagram and classified as a member of the TDY group.
Mentions: A BLAST search (http://www.ncbi.nlm.nih.gov/BLAST) and multi-alignment analysis revealed that GhMPK16 is highly related to other group D MAPKs, sharing a homology of 81.48% to AtMPK16, 72.18% to TaMAPK2, 74.56% to OsMPK16-1 and 76.02% to ZmMPK6 (Figure 1A). Moreover, as with other known plant group D MAPKs, GhMPK16 has the same family signature of 11 conserved subdomains, an activation loop (T-loop), a phosphorylation motif (TDY motif) in the T-loop, and an extended C-terminal region relative to groups A, B and C. Additionally, GhMPK16 also lacks a C-terminal CD domain, a feature conserved in MAPKs belonging to the other groups (A, B, C) [4].

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