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A receptor-like kinase gene (GbRLK) from Gossypium barbadense enhances salinity and drought-stress tolerance in Arabidopsis.

Zhao J, Gao Y, Zhang Z, Chen T, Guo W, Zhang T - BMC Plant Biol. (2013)

Bottom Line: Transgenic Arabidopsis with constitutive overexpression of GbRLK exhibited a reduced rate of water loss in leaves in vitro, along with improved salinity and drought tolerance and increased sensitivity to ABA compared with non-transgenic Col-0 Arabidopsis.Overexpression of GbRLK may improve stress tolerance by regulating stress-responsive genes to reduce water loss.Further studying of GbRLK will help elucidate abiotic stress signaling pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Key Laboratory of Crop Genetics & Germplasm Enhancement, MOE Hybrid Cotton R&D Engineering Research Center, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China.

ABSTRACT

Background: Cotton (Gossypium spp.) is widely cultivated due to the important economic value of its fiber. However, extreme environmental degradation impedes cotton growth and production. Receptor-like kinase (RLK) proteins play important roles in signal transduction and participate in a diverse range of processes in response to plant hormones and environmental cues. Here, we introduced an RLK gene (GbRLK) from cotton into Arabidopsis and investigated its role in imparting abiotic stress tolerance.

Results: GbRLK transcription was induced by exogenously supplied abscisic acid (ABA), salicylic acid, methyl jasmonate, mock drought conditions and high salinity. We cloned the promoter sequence of this gene via self-formed adaptor PCR. Sequence analysis revealed that the promoter region contains many cis-acting stress-responsive elements such as ABRE, W-Box, MYB-core, W-Box core, TCA-element and others. We constructed a vector containing a 1,890-bp sequence in the 5' region upstream of the initiation codon of this promoter and transformed it into Arabidopsis thaliana. GUS histochemical staining analysis showed that GbRLK was expressed mainly in leaf veins, petioles and roots of transgenic Arabidopsis, but not in the cotyledons or root hairs. GbRLK promoter activity was induced by ABA, PEG, NaCl and Verticillium dahliae. Transgenic Arabidopsis with constitutive overexpression of GbRLK exhibited a reduced rate of water loss in leaves in vitro, along with improved salinity and drought tolerance and increased sensitivity to ABA compared with non-transgenic Col-0 Arabidopsis. Expression analysis of stress-responsive genes in GbRLK Arabidopsis revealed that there was increased expression of genes involved in the ABA-dependent signaling pathway (AtRD20, AtRD22 and AtRD26) and antioxidant genes (AtCAT1, AtCCS, AtCSD2 and AtCSD1) but not ion transporter genes (AtNHX1, AtSOS1).

Conclusions: GbRLK is involved in the drought and high salinity stresses pathway by activating or participating in the ABA signaling pathway. Overexpression of GbRLK may improve stress tolerance by regulating stress-responsive genes to reduce water loss. GbRLK may be employed in the genetic engineering of novel cotton cultivars in the future. Further studying of GbRLK will help elucidate abiotic stress signaling pathways.

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Phylogenetic analysis of GbRLK. (a) Alignment of deduced amino acid sequences of GbRLK and other kinases. (AAB47421, Pto from tomato; BAB08490, protein kinase from Arabidopsis; AAT70497, protein kinase from peach; BAD37843, protein kinase from rice; AAD44031, PK3 from barley; AAK20741, TAK33 from barley; AAB82755, Xa21 from rice). (b) Phylogenetic relationships between GbRLK and other kinase proteins from Gossypium hirsutum and Gossypium barbadense. (c) Phylogenetic relationships between GbRLK and other reported receptor-like kinase proteins from the eudicot Arabidopsis thaliana (AtANXUR1, AEE74120; AtPBS1, AED91858; AtCERK1, AEE76532; AtPR5K, AED94289; AtACR4, AEE79920; AtWAK1, AEE30079; AtRKF3, AEC10923; AtBRI1, AEE87069; AtERECTA, AEC07825; AtCLV1, AEE35762; AtRLK5, AEE85494; AtPERK1, AEE76920; AtPEK12, AEE30401; AtCRK1, AEE29801; AtSD113, AEE28721; AtRKS1, AEE28720; AtCRK5, EFH43988; AtCRK10, AEE84719; AtRLK3, CAA09731; AtFLS2, AED95370; AtLRK13, AEE78027; AtRLK1, AED97394; AtRLK4, BAC42412; AT4G27300; AT4G11890; AT5G24080; AT4G32300; AT5G01540; AT4GO2410; AT3G59750; AT1G09970; AT1G66880; AT2G33580) and some representative members of Oryza sativa (Xa21, AAB82755), Lycopersicon esculentum (LePTO, BAB08490), and Triticum aestivum (TaLRK10, AAC49629). The phylogenetic tree was constructed using the maximum likelihood method in MEGA 5.0. Numbers on internal nodes are the percentage bootstrap support values (1000 re-sampling); only values exceeding 50% are shown.
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Figure 1: Phylogenetic analysis of GbRLK. (a) Alignment of deduced amino acid sequences of GbRLK and other kinases. (AAB47421, Pto from tomato; BAB08490, protein kinase from Arabidopsis; AAT70497, protein kinase from peach; BAD37843, protein kinase from rice; AAD44031, PK3 from barley; AAK20741, TAK33 from barley; AAB82755, Xa21 from rice). (b) Phylogenetic relationships between GbRLK and other kinase proteins from Gossypium hirsutum and Gossypium barbadense. (c) Phylogenetic relationships between GbRLK and other reported receptor-like kinase proteins from the eudicot Arabidopsis thaliana (AtANXUR1, AEE74120; AtPBS1, AED91858; AtCERK1, AEE76532; AtPR5K, AED94289; AtACR4, AEE79920; AtWAK1, AEE30079; AtRKF3, AEC10923; AtBRI1, AEE87069; AtERECTA, AEC07825; AtCLV1, AEE35762; AtRLK5, AEE85494; AtPERK1, AEE76920; AtPEK12, AEE30401; AtCRK1, AEE29801; AtSD113, AEE28721; AtRKS1, AEE28720; AtCRK5, EFH43988; AtCRK10, AEE84719; AtRLK3, CAA09731; AtFLS2, AED95370; AtLRK13, AEE78027; AtRLK1, AED97394; AtRLK4, BAC42412; AT4G27300; AT4G11890; AT5G24080; AT4G32300; AT5G01540; AT4GO2410; AT3G59750; AT1G09970; AT1G66880; AT2G33580) and some representative members of Oryza sativa (Xa21, AAB82755), Lycopersicon esculentum (LePTO, BAB08490), and Triticum aestivum (TaLRK10, AAC49629). The phylogenetic tree was constructed using the maximum likelihood method in MEGA 5.0. Numbers on internal nodes are the percentage bootstrap support values (1000 re-sampling); only values exceeding 50% are shown.

Mentions: A functional GbRLK gene [NCBI accession number KC422776], which was induced by Verticillium dahliae, was previously identified among a group of expressed resistance gene analogs from the disease-resistant cotton Gossypium barbasense cv. Hai7124 using RACE[32]. The cDNA was 1536-bp in length and the predicted ORF starts at nucleotide 330 and ends at nucleotide 1409, encoding a predicted protein of 359 amino acids. BLASTx searches revealed that the predicted product shared 68% identity and 75% similarity with the soybean G-type lectin S-receptor-like serine/threonine -protein kinase (E value: 0), 56% identity and 72% similarity with the rice PK3 (E value: 1e-98), 48% and 65% (E value: 4e-73) with the wheat TAK33, and 48% and 64% (E value: 2e-71) with the barley receptor-like protein kinase, respectively. The protein possessed intact signature features of protein tyrosine kinases, and contained conserved motifs characteristic of I-XI of receptor-like protein kinases, which are possessed in the majority of protein kinases. Multiple sequence alignments using BLASTx revealed that the protein shared homology in other regions with protein kinases from various other crop species (Figure 1a). We therefore conclude that this protein may belong to the receptor-like protein kinase family and be a member of the RLK gene family. This gene was thereafter designated as G. barbadense receptor-like protein kinase (GbRLK). GbRLK contains an ATP binding site (residues 32 to 40), a D activation site (at residue 150), four N-glycosylation sites, four N-myristoylation sites, three C terminal phosphorylation sites, one amidation site, four tyrosine kinase II phosphorylation sites, and a membrane-spanning region (residues 28–46).


A receptor-like kinase gene (GbRLK) from Gossypium barbadense enhances salinity and drought-stress tolerance in Arabidopsis.

Zhao J, Gao Y, Zhang Z, Chen T, Guo W, Zhang T - BMC Plant Biol. (2013)

Phylogenetic analysis of GbRLK. (a) Alignment of deduced amino acid sequences of GbRLK and other kinases. (AAB47421, Pto from tomato; BAB08490, protein kinase from Arabidopsis; AAT70497, protein kinase from peach; BAD37843, protein kinase from rice; AAD44031, PK3 from barley; AAK20741, TAK33 from barley; AAB82755, Xa21 from rice). (b) Phylogenetic relationships between GbRLK and other kinase proteins from Gossypium hirsutum and Gossypium barbadense. (c) Phylogenetic relationships between GbRLK and other reported receptor-like kinase proteins from the eudicot Arabidopsis thaliana (AtANXUR1, AEE74120; AtPBS1, AED91858; AtCERK1, AEE76532; AtPR5K, AED94289; AtACR4, AEE79920; AtWAK1, AEE30079; AtRKF3, AEC10923; AtBRI1, AEE87069; AtERECTA, AEC07825; AtCLV1, AEE35762; AtRLK5, AEE85494; AtPERK1, AEE76920; AtPEK12, AEE30401; AtCRK1, AEE29801; AtSD113, AEE28721; AtRKS1, AEE28720; AtCRK5, EFH43988; AtCRK10, AEE84719; AtRLK3, CAA09731; AtFLS2, AED95370; AtLRK13, AEE78027; AtRLK1, AED97394; AtRLK4, BAC42412; AT4G27300; AT4G11890; AT5G24080; AT4G32300; AT5G01540; AT4GO2410; AT3G59750; AT1G09970; AT1G66880; AT2G33580) and some representative members of Oryza sativa (Xa21, AAB82755), Lycopersicon esculentum (LePTO, BAB08490), and Triticum aestivum (TaLRK10, AAC49629). The phylogenetic tree was constructed using the maximum likelihood method in MEGA 5.0. Numbers on internal nodes are the percentage bootstrap support values (1000 re-sampling); only values exceeding 50% are shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 1: Phylogenetic analysis of GbRLK. (a) Alignment of deduced amino acid sequences of GbRLK and other kinases. (AAB47421, Pto from tomato; BAB08490, protein kinase from Arabidopsis; AAT70497, protein kinase from peach; BAD37843, protein kinase from rice; AAD44031, PK3 from barley; AAK20741, TAK33 from barley; AAB82755, Xa21 from rice). (b) Phylogenetic relationships between GbRLK and other kinase proteins from Gossypium hirsutum and Gossypium barbadense. (c) Phylogenetic relationships between GbRLK and other reported receptor-like kinase proteins from the eudicot Arabidopsis thaliana (AtANXUR1, AEE74120; AtPBS1, AED91858; AtCERK1, AEE76532; AtPR5K, AED94289; AtACR4, AEE79920; AtWAK1, AEE30079; AtRKF3, AEC10923; AtBRI1, AEE87069; AtERECTA, AEC07825; AtCLV1, AEE35762; AtRLK5, AEE85494; AtPERK1, AEE76920; AtPEK12, AEE30401; AtCRK1, AEE29801; AtSD113, AEE28721; AtRKS1, AEE28720; AtCRK5, EFH43988; AtCRK10, AEE84719; AtRLK3, CAA09731; AtFLS2, AED95370; AtLRK13, AEE78027; AtRLK1, AED97394; AtRLK4, BAC42412; AT4G27300; AT4G11890; AT5G24080; AT4G32300; AT5G01540; AT4GO2410; AT3G59750; AT1G09970; AT1G66880; AT2G33580) and some representative members of Oryza sativa (Xa21, AAB82755), Lycopersicon esculentum (LePTO, BAB08490), and Triticum aestivum (TaLRK10, AAC49629). The phylogenetic tree was constructed using the maximum likelihood method in MEGA 5.0. Numbers on internal nodes are the percentage bootstrap support values (1000 re-sampling); only values exceeding 50% are shown.
Mentions: A functional GbRLK gene [NCBI accession number KC422776], which was induced by Verticillium dahliae, was previously identified among a group of expressed resistance gene analogs from the disease-resistant cotton Gossypium barbasense cv. Hai7124 using RACE[32]. The cDNA was 1536-bp in length and the predicted ORF starts at nucleotide 330 and ends at nucleotide 1409, encoding a predicted protein of 359 amino acids. BLASTx searches revealed that the predicted product shared 68% identity and 75% similarity with the soybean G-type lectin S-receptor-like serine/threonine -protein kinase (E value: 0), 56% identity and 72% similarity with the rice PK3 (E value: 1e-98), 48% and 65% (E value: 4e-73) with the wheat TAK33, and 48% and 64% (E value: 2e-71) with the barley receptor-like protein kinase, respectively. The protein possessed intact signature features of protein tyrosine kinases, and contained conserved motifs characteristic of I-XI of receptor-like protein kinases, which are possessed in the majority of protein kinases. Multiple sequence alignments using BLASTx revealed that the protein shared homology in other regions with protein kinases from various other crop species (Figure 1a). We therefore conclude that this protein may belong to the receptor-like protein kinase family and be a member of the RLK gene family. This gene was thereafter designated as G. barbadense receptor-like protein kinase (GbRLK). GbRLK contains an ATP binding site (residues 32 to 40), a D activation site (at residue 150), four N-glycosylation sites, four N-myristoylation sites, three C terminal phosphorylation sites, one amidation site, four tyrosine kinase II phosphorylation sites, and a membrane-spanning region (residues 28–46).

Bottom Line: Transgenic Arabidopsis with constitutive overexpression of GbRLK exhibited a reduced rate of water loss in leaves in vitro, along with improved salinity and drought tolerance and increased sensitivity to ABA compared with non-transgenic Col-0 Arabidopsis.Overexpression of GbRLK may improve stress tolerance by regulating stress-responsive genes to reduce water loss.Further studying of GbRLK will help elucidate abiotic stress signaling pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Key Laboratory of Crop Genetics & Germplasm Enhancement, MOE Hybrid Cotton R&D Engineering Research Center, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China.

ABSTRACT

Background: Cotton (Gossypium spp.) is widely cultivated due to the important economic value of its fiber. However, extreme environmental degradation impedes cotton growth and production. Receptor-like kinase (RLK) proteins play important roles in signal transduction and participate in a diverse range of processes in response to plant hormones and environmental cues. Here, we introduced an RLK gene (GbRLK) from cotton into Arabidopsis and investigated its role in imparting abiotic stress tolerance.

Results: GbRLK transcription was induced by exogenously supplied abscisic acid (ABA), salicylic acid, methyl jasmonate, mock drought conditions and high salinity. We cloned the promoter sequence of this gene via self-formed adaptor PCR. Sequence analysis revealed that the promoter region contains many cis-acting stress-responsive elements such as ABRE, W-Box, MYB-core, W-Box core, TCA-element and others. We constructed a vector containing a 1,890-bp sequence in the 5' region upstream of the initiation codon of this promoter and transformed it into Arabidopsis thaliana. GUS histochemical staining analysis showed that GbRLK was expressed mainly in leaf veins, petioles and roots of transgenic Arabidopsis, but not in the cotyledons or root hairs. GbRLK promoter activity was induced by ABA, PEG, NaCl and Verticillium dahliae. Transgenic Arabidopsis with constitutive overexpression of GbRLK exhibited a reduced rate of water loss in leaves in vitro, along with improved salinity and drought tolerance and increased sensitivity to ABA compared with non-transgenic Col-0 Arabidopsis. Expression analysis of stress-responsive genes in GbRLK Arabidopsis revealed that there was increased expression of genes involved in the ABA-dependent signaling pathway (AtRD20, AtRD22 and AtRD26) and antioxidant genes (AtCAT1, AtCCS, AtCSD2 and AtCSD1) but not ion transporter genes (AtNHX1, AtSOS1).

Conclusions: GbRLK is involved in the drought and high salinity stresses pathway by activating or participating in the ABA signaling pathway. Overexpression of GbRLK may improve stress tolerance by regulating stress-responsive genes to reduce water loss. GbRLK may be employed in the genetic engineering of novel cotton cultivars in the future. Further studying of GbRLK will help elucidate abiotic stress signaling pathways.

Show MeSH
Related in: MedlinePlus