Karam Singh - Academia.edu (original) (raw)

Papers by Karam Singh

Functional Plant Biology, 2005

Plant Journal, 2000

Dof proteins are unique to plants and contain a single zinc finger DNA-binding domain called the ... more Dof proteins are unique to plants and contain a single zinc finger DNA-binding domain called the Dof domain. OBP1, an Arabidopsis Dof protein, was previously isolated through an interaction with OBF4, an ocs element-binding protein. Two additional Dof proteins, called OBP2 and OBP3, were isolated through homologous screening. All three OBP proteins contain transcriptional activation domains in their C-terminal region. While no significant differences were found between the OBP proteins in terms of their DNA and protein-binding properties, tissue-specific RNA expression patterns were found. The RNA expression levels of all three OBP proteins increased following treatment with auxin, salicylic acid (SA) or cycloheximide, although the level of induction varied among the different proteins and in the tissues tested. These results suggest that even though they have similar DNA binding and protein–protein interaction properties, the different OBP proteins are likely to have distinct functions in specific parts of the plant. There is a good correlation between the expression of the OBP proteins and the ocs element, a stress-response element which is also induced by auxin, SA and cycloheximide. To begin to analyze the function of the OBP proteins, transgenic lines overexpressing OBP3 were generated. These plants showed a severe growth defect with altered root development and yellowish leaves. The severity of the growth defects correlated with OBP3 expression levels and in some cases led to death, suggesting that some Dof proteins play important roles in plant growth and development.

Plant Journal, 2009

Plant glutathione transferases (GSTs) are induced by diverse biotic and abiotic stimuli, and are ... more Plant glutathione transferases (GSTs) are induced by diverse biotic and abiotic stimuli, and are important for protecting plants against oxidative damage. We have studied the primary transcriptional stress response of the entire Arabidopsis GST family to seven stresses, including both biotic and abiotic stimuli, with a focus on early changes in gene expression. Our results indicate that individual GST genes are highly specific in their induction patterns. Furthermore, we have been able to link individual GSTs to particular stress stimuli. Using RNAi, we successfully co-silenced a group of four phi GSTs that represent some of the most highly expressed GST genes. Despite a marked reduction in total phi GST protein levels, the transgenic plants showed no reduction in GST activity as measured using the model substrate 1-chloro-2,4-dinitrobenzene (CDNB), and appeared to have surprisingly robust physical phenotypes during stress. However, analysis of metabolite pools showed oxidation of the glutathione pool in the RNAi lines, and we observed alterations in carbon and nitrogen compounds following salicylic acid and hydrogen peroxide stress treatments, indicative of oxidative modification of primary metabolism. Thus, there appears to be a high degree of functional redundancy within the Arabidopsis GST family, with extensive disruption being required to reveal the roles of phi GSTs in protection against oxidative stress.

Plant Journal, 1993

Ocs-elements, a family of 20 bp DNA sequences, are components of a number of promoters active in ... more Ocs-elements, a family of 20 bp DNA sequences, are components of a number of promoters active in plants. In the maize BMS cell line the dominant ocs-element binding activity is the ocs-element transcription factor complex called OTF. The isolation of cDNA clones from a BMS cDNA expression library for two bZIP (basic region-leucine zipper) proteins that bind the ocs-element sequence and are good candidates for forming at least part of OTF is described. The two ocs-element binding proteins, called OBF3.1 and OBF3.2, are closely related, with the OBF3.1 protein sharing 95.8% amino acid homology with part of the OBF3.2 protein although there were significant differences in the 3′ untranslated regions. Genomic Southern blot analysis revealed a small gene family with a minimum of two OBF3 loci mapping to chromosomes 3L 105 and 8L075. The OBF3.1 protein shared considerable homology with the wheat HBP1b protein (80% amino acid identity) and to a lesser extent with the tobacco TGA1a protein. OBF3.1 like HBP1b was able to bind well to the Hex sequence but poorly to G-box/ABRE sequences. Interestingly, OBF3.1 bound eightfold more efficiently to an ocs-element sequence than TGA1a, raising the possibility that OBF3.1 and TGA1a may be distinct members of an OBF3/TGA subfamily.

Functional Plant Biology, 2005

New Phytologist, 2006

• Host resistance to aphids is poorly understood. Medicago truncatula, a model legume and cultiva... more • Host resistance to aphids is poorly understood. Medicago truncatula, a model legume and cultivated pasture species, was used to elucidate defense against two aphid species, Therioaphis trifolii f. maculata (spotted alfalfa aphid, SAA) and Acyrthosiphon kondoi (bluegreen aphid, BGA).• Aphid performance and plant damage were compared between near-isogenic cultivars, Mogul and Borung, that differ in resistance to both aphids. Analyses of aphid resistance in Mogul × Borung F2 plants and their progeny revealed modes of action and chromosome locations of resistance genes.• Separate genes were identified for SAA resistance (TTR) and BGA resistance (AKR); both mapped to chromosome 3 but were found to act independently to reduce survival and growth of their target aphid species. The TTR locus controls distinct, and contrasting, local and systemic plant responses between the near-isogenic cultivars.• TTR-mediated plant responses imply interaction between a resistance factor(s) in vascular tissue and a bioactive component(s) of SAA saliva. Features of both resistance traits suggest homology to aphid resistance in other legumes; elucidation of their molecular mechanisms will likely apply to other aphid–plant interactions.Host resistance to aphids is poorly understood. Medicago truncatula, a model legume and cultivated pasture species, was used to elucidate defense against two aphid species, Therioaphis trifolii f. maculata (spotted alfalfa aphid, SAA) and Acyrthosiphon kondoi (bluegreen aphid, BGA).Aphid performance and plant damage were compared between near-isogenic cultivars, Mogul and Borung, that differ in resistance to both aphids. Analyses of aphid resistance in Mogul × Borung F2 plants and their progeny revealed modes of action and chromosome locations of resistance genes.Separate genes were identified for SAA resistance (TTR) and BGA resistance (AKR); both mapped to chromosome 3 but were found to act independently to reduce survival and growth of their target aphid species. The TTR locus controls distinct, and contrasting, local and systemic plant responses between the near-isogenic cultivars.TTR-mediated plant responses imply interaction between a resistance factor(s) in vascular tissue and a bioactive component(s) of SAA saliva. Features of both resistance traits suggest homology to aphid resistance in other legumes; elucidation of their molecular mechanisms will likely apply to other aphid–plant interactions.

Current Opinion in Plant Biology, 2002

Proceedings of The National Academy of Sciences, 1994

Plant Molecular Biology, 2004

Plant glutathione S-transferases (GSTs) are a large group of multifunctional proteins that are in... more Plant glutathione S-transferases (GSTs) are a large group of multifunctional proteins that are induced by diverse stimuli. Using proteomic approaches we identified 20 GSTs at the protein level in Arabidopsis cell culture with a combination of GST antibody detection, LC-MS/MS analysis of 23–30 kDa proteins and glutathione-affinity chromatography. GSTs identified were from phi, tau, theta, zeta and DHAR sub-sections of the GST superfamily of 53 members. We have uncovered preliminary evidence for post-translational modifications of plant GSTs and show that phosphorylation is unlikely to be responsible. Detailed analysis of GST expression in response to treatment with 0.01–1 mM of the plant defence signal salicylic acid (SA) uncovered some interesting features. Firstly, GSTs appear to display class-specific concentration-dependent SA induction profiles highlighting differences between the large, plant specific phi and tau classes. Secondly, different members of the same class, while sharing similar SA dose responses, may display differences in terms of magnitude and timing of induction, further highlighting the breadth of GST gene regulation. Thirdly, closely related members of the same class (GSTF6 and GSTF7), arising via tandem duplication, may be regulated differently in terms of basal expression levels and also magnitude of induction raising questions about the role of subfunctionalisation within this family. Our results reveal that GSTs exhibit class specific responses to SA treatment suggesting that several mechanisms are acting to induce GSTs upon SA treatment and hinting at class-specific functions for this large and important, yet still relatively elusive gene family.

Plant Molecular Biology, 1997

In animals and fungi, small cysteine-rich proteins called metallothioneins (MTs) play a role in h... more In animals and fungi, small cysteine-rich proteins called metallothioneins (MTs) play a role in heavy metal tolerance. MT genes have been isolated in plants, but their function remains to be elucidated. We have isolated two distinct Vicia faba MT genes that belong to the type 1 group of plant MT genes in contrast to a MT gene we previously isolated that belongs to type 2. We found similarities and differences between the V. faba MT genes. The RNA expression patterns differed and this was most pronounced in roots, which contained high MT1 but very low MT2 RNA levels. Like MT2, MT1 transcript levels were not significantly affected by treatment with Cd, Cu, Fe and Zn, at least under the experimental conditions. MT RNA levels varied in leaves and stem internodes of different developmental ages, with the highest expression in the older tissue. The levels of MT RNA correlated inversely with endogenous Cd, Cu and Fe levels within different internodes, but not with a number of other metals tested (including Zn). The three bean MTs were expressed in Escherichia coli and found to bind Cd, Cu and Zn but not to Fe. The MTs were tested to determine if they differed in their ability to bind a specific metal but no significant differences in binding were observed.

Plant Molecular Biology, 1994

Animal metallothioneins (MTs) are cysteine-rich, low-molecular-weight proteins that bind to heavy... more Animal metallothioneins (MTs) are cysteine-rich, low-molecular-weight proteins that bind to heavy metals and are believed to play a role in their metabolism and detoxification. Genes encoding MT-like proteins have been isolated in a number of plants although their function remains to be elucidated. We describe the isolation and characterization of a bean cDNA encoding an MT-like protein. The bean gene, called MT, was isolated as a result of a differential screen for genes that are expressed in leaves but not in the most common cell type, the mesophyll cell. MT contained two regions with abundant cysteines and sequence comparison found that MT had greatest homology to MT-like subtype 2 from other plant species. Northern blot analysis demonstrated that MT was expressed in the leaf, stem and flower, at very low levels in roots and was not detectable in mesophyll protoplasts. MT transcript levels were not significantly affected by treatment with Cu, Zn or Cd. In the leaf, in situ hybridization studies demonstrated striking cell specificity with MT expression confined predominantly to trichomes. Possible explanations for the pronounced expression of MT in leaf trichomes are discussed.

Molecular Plant-microbe Interactions, 2007

Aphids are major insect pests of plants that feed directly from the phloem. We used the model leg... more Aphids are major insect pests of plants that feed directly from the phloem. We used the model legume Medicago truncatula Gaert. (barrel medic) to elucidate host resistance to aphids and identified a single dominant gene which confers resistance to Acyrthosiphon kondoi Shinji (bluegreen aphid). To understand how this gene conditions resistance to bluegreen aphid, transcription profiling of 23 defense-related genes representing various signaling pathways was undertaken using a pair of near-isogenic lines that are susceptible or resistant to bluegreen aphid. All salicylic acid- and ethylene-responsive genes tested were induced by bluegreen aphid in resistant and susceptible plants, although there were some differences in the magnitude and kinetics of the induction. In contrast, 10 of 13 genes associated with the octadecanoid pathway were induced exclusively in the resistant plants following bluegreen aphid infestation. These results are in contrast to plant-pathogen interactions where similar sets of defense genes typically are induced in compatible interactions, but to a lesser degree and later than in incompatible interactions. Treatment of susceptible plants with methyl jasmonate reduced bluegreen aphid infestation but not to the same levels as the resistant line. Together, these results strongly suggest that the octadecanoid pathway is important for this naturally derived aphid resistance trait.

Plant Journal, 1993

Ocs-elements are a group of related, bipartite promoter elements which have been exploited by two... more Ocs-elements are a group of related, bipartite promoter elements which have been exploited by two distinct groups of plant pathogens, Agrobacterium and certain viruses to express genes in plants. The genes for two Arabidopsis bZIP (basic region-leucine zipper) proteins that bind to ocs-elements have been isolated and characterized. The genes, called OBF4 and OBF5, were isolated by screening an Arabidopsis genomic library with degenerate oligonucleotides complementary to the DNA-binding domains of other plant ocs-element-binding proteins. The OBF4 and OBF5 proteins show 53% amino acid identity but low DNA homology. Southern blot analysis demonstrated that each of the OBF genes is a member of a small family. OBF4 is more similar to the tobacco TGA1a and Arabidopsis TGA1 proteins, while OBF5 is more similar to the maize OBF3.1, wheat HBP1b and Arabidopsis aHBP1b proteins. The DNA-binding properties of OBF4 and OBF5 were similar although OBF5 was able to bind simultaneously to both halves of the ocs-element more efficiently than OBF4. This difference in binding to the ocs-element between two closely related proteins from the same species is potentially significant since binding to both halves of the ocs-element is a pre-requisite for in vivo transcriptional activity.

Plant Journal, 2003

Overexpression of a salicylic-acid (SA)-inducible Arabidopsis DNA binding with one finger (Dof) t... more Overexpression of a salicylic-acid (SA)-inducible Arabidopsis DNA binding with one finger (Dof) transcription factor, called OBF-binding protein 3 (OBP3; AtDof3.6), has previously been shown to result in growth defects. In this study, suppressive subtraction hybridization (SSH) was used to isolate genes induced in an OBP3-overexpression line and several putative clones, called OBP3-responsive genes (ORGs), were isolated. The link with the induced expression levels of these genes and OBP3 overexpression was confirmed by analysing additional OBP3-overexpression lines. ORG1 through ORG4 are novel genes, while ORG5 is an extensin gene, AtExt1. While ORG4 has no similarity with other proteins in the database, ORG1 has weak similarity in different regions of the predicted protein with CDC2 and fibrillin. ORG2 and ORG3 share 80% overall identity in their deduced amino acid sequences and contain a basic helix-loop-helix DNA-binding domain, suggesting that ORG2 and ORG3 may be transcription factors. The expression of the ORG1, ORG2 and ORG3 genes was co-regulated under all conditions examined including upregulation by SA and downregulation by jasmonic acid (JA). Fifteen OBP3-silenced lines were generated to further explore the function of OBP3. Although there were no visible phenotypic changes in any of these lines, the expression of ORG1, ORG2 and ORG3 was reduced. Among the ORG genes, ORG1, ORG2 and ORG3 contained the highest number of potential Dof-binding sites in the promoter region, and their expression was significantly increased within 3 h after induction of OBP3 expression using an inducible promoter system, and closely reflected the expression levels of the exogenous OBP3 protein. The results from the gain-of-function and loss-of-function experiments suggest that the ORG1, ORG2 and ORG3 genes are direct target genes of OBP3.

Functional Plant Biology, 2005

Plant Journal, 2000

Dof proteins are unique to plants and contain a single zinc finger DNA-binding domain called the ... more Dof proteins are unique to plants and contain a single zinc finger DNA-binding domain called the Dof domain. OBP1, an Arabidopsis Dof protein, was previously isolated through an interaction with OBF4, an ocs element-binding protein. Two additional Dof proteins, called OBP2 and OBP3, were isolated through homologous screening. All three OBP proteins contain transcriptional activation domains in their C-terminal region. While no significant differences were found between the OBP proteins in terms of their DNA and protein-binding properties, tissue-specific RNA expression patterns were found. The RNA expression levels of all three OBP proteins increased following treatment with auxin, salicylic acid (SA) or cycloheximide, although the level of induction varied among the different proteins and in the tissues tested. These results suggest that even though they have similar DNA binding and protein–protein interaction properties, the different OBP proteins are likely to have distinct functions in specific parts of the plant. There is a good correlation between the expression of the OBP proteins and the ocs element, a stress-response element which is also induced by auxin, SA and cycloheximide. To begin to analyze the function of the OBP proteins, transgenic lines overexpressing OBP3 were generated. These plants showed a severe growth defect with altered root development and yellowish leaves. The severity of the growth defects correlated with OBP3 expression levels and in some cases led to death, suggesting that some Dof proteins play important roles in plant growth and development.

Plant Journal, 2009

Plant glutathione transferases (GSTs) are induced by diverse biotic and abiotic stimuli, and are ... more Plant glutathione transferases (GSTs) are induced by diverse biotic and abiotic stimuli, and are important for protecting plants against oxidative damage. We have studied the primary transcriptional stress response of the entire Arabidopsis GST family to seven stresses, including both biotic and abiotic stimuli, with a focus on early changes in gene expression. Our results indicate that individual GST genes are highly specific in their induction patterns. Furthermore, we have been able to link individual GSTs to particular stress stimuli. Using RNAi, we successfully co-silenced a group of four phi GSTs that represent some of the most highly expressed GST genes. Despite a marked reduction in total phi GST protein levels, the transgenic plants showed no reduction in GST activity as measured using the model substrate 1-chloro-2,4-dinitrobenzene (CDNB), and appeared to have surprisingly robust physical phenotypes during stress. However, analysis of metabolite pools showed oxidation of the glutathione pool in the RNAi lines, and we observed alterations in carbon and nitrogen compounds following salicylic acid and hydrogen peroxide stress treatments, indicative of oxidative modification of primary metabolism. Thus, there appears to be a high degree of functional redundancy within the Arabidopsis GST family, with extensive disruption being required to reveal the roles of phi GSTs in protection against oxidative stress.

Plant Journal, 1993

Ocs-elements, a family of 20 bp DNA sequences, are components of a number of promoters active in ... more Ocs-elements, a family of 20 bp DNA sequences, are components of a number of promoters active in plants. In the maize BMS cell line the dominant ocs-element binding activity is the ocs-element transcription factor complex called OTF. The isolation of cDNA clones from a BMS cDNA expression library for two bZIP (basic region-leucine zipper) proteins that bind the ocs-element sequence and are good candidates for forming at least part of OTF is described. The two ocs-element binding proteins, called OBF3.1 and OBF3.2, are closely related, with the OBF3.1 protein sharing 95.8% amino acid homology with part of the OBF3.2 protein although there were significant differences in the 3′ untranslated regions. Genomic Southern blot analysis revealed a small gene family with a minimum of two OBF3 loci mapping to chromosomes 3L 105 and 8L075. The OBF3.1 protein shared considerable homology with the wheat HBP1b protein (80% amino acid identity) and to a lesser extent with the tobacco TGA1a protein. OBF3.1 like HBP1b was able to bind well to the Hex sequence but poorly to G-box/ABRE sequences. Interestingly, OBF3.1 bound eightfold more efficiently to an ocs-element sequence than TGA1a, raising the possibility that OBF3.1 and TGA1a may be distinct members of an OBF3/TGA subfamily.

Functional Plant Biology, 2005

New Phytologist, 2006

• Host resistance to aphids is poorly understood. Medicago truncatula, a model legume and cultiva... more • Host resistance to aphids is poorly understood. Medicago truncatula, a model legume and cultivated pasture species, was used to elucidate defense against two aphid species, Therioaphis trifolii f. maculata (spotted alfalfa aphid, SAA) and Acyrthosiphon kondoi (bluegreen aphid, BGA).• Aphid performance and plant damage were compared between near-isogenic cultivars, Mogul and Borung, that differ in resistance to both aphids. Analyses of aphid resistance in Mogul × Borung F2 plants and their progeny revealed modes of action and chromosome locations of resistance genes.• Separate genes were identified for SAA resistance (TTR) and BGA resistance (AKR); both mapped to chromosome 3 but were found to act independently to reduce survival and growth of their target aphid species. The TTR locus controls distinct, and contrasting, local and systemic plant responses between the near-isogenic cultivars.• TTR-mediated plant responses imply interaction between a resistance factor(s) in vascular tissue and a bioactive component(s) of SAA saliva. Features of both resistance traits suggest homology to aphid resistance in other legumes; elucidation of their molecular mechanisms will likely apply to other aphid–plant interactions.Host resistance to aphids is poorly understood. Medicago truncatula, a model legume and cultivated pasture species, was used to elucidate defense against two aphid species, Therioaphis trifolii f. maculata (spotted alfalfa aphid, SAA) and Acyrthosiphon kondoi (bluegreen aphid, BGA).Aphid performance and plant damage were compared between near-isogenic cultivars, Mogul and Borung, that differ in resistance to both aphids. Analyses of aphid resistance in Mogul × Borung F2 plants and their progeny revealed modes of action and chromosome locations of resistance genes.Separate genes were identified for SAA resistance (TTR) and BGA resistance (AKR); both mapped to chromosome 3 but were found to act independently to reduce survival and growth of their target aphid species. The TTR locus controls distinct, and contrasting, local and systemic plant responses between the near-isogenic cultivars.TTR-mediated plant responses imply interaction between a resistance factor(s) in vascular tissue and a bioactive component(s) of SAA saliva. Features of both resistance traits suggest homology to aphid resistance in other legumes; elucidation of their molecular mechanisms will likely apply to other aphid–plant interactions.

Current Opinion in Plant Biology, 2002

Proceedings of The National Academy of Sciences, 1994

Plant Molecular Biology, 2004

Plant glutathione S-transferases (GSTs) are a large group of multifunctional proteins that are in... more Plant glutathione S-transferases (GSTs) are a large group of multifunctional proteins that are induced by diverse stimuli. Using proteomic approaches we identified 20 GSTs at the protein level in Arabidopsis cell culture with a combination of GST antibody detection, LC-MS/MS analysis of 23–30 kDa proteins and glutathione-affinity chromatography. GSTs identified were from phi, tau, theta, zeta and DHAR sub-sections of the GST superfamily of 53 members. We have uncovered preliminary evidence for post-translational modifications of plant GSTs and show that phosphorylation is unlikely to be responsible. Detailed analysis of GST expression in response to treatment with 0.01–1 mM of the plant defence signal salicylic acid (SA) uncovered some interesting features. Firstly, GSTs appear to display class-specific concentration-dependent SA induction profiles highlighting differences between the large, plant specific phi and tau classes. Secondly, different members of the same class, while sharing similar SA dose responses, may display differences in terms of magnitude and timing of induction, further highlighting the breadth of GST gene regulation. Thirdly, closely related members of the same class (GSTF6 and GSTF7), arising via tandem duplication, may be regulated differently in terms of basal expression levels and also magnitude of induction raising questions about the role of subfunctionalisation within this family. Our results reveal that GSTs exhibit class specific responses to SA treatment suggesting that several mechanisms are acting to induce GSTs upon SA treatment and hinting at class-specific functions for this large and important, yet still relatively elusive gene family.

Plant Molecular Biology, 1997

In animals and fungi, small cysteine-rich proteins called metallothioneins (MTs) play a role in h... more In animals and fungi, small cysteine-rich proteins called metallothioneins (MTs) play a role in heavy metal tolerance. MT genes have been isolated in plants, but their function remains to be elucidated. We have isolated two distinct Vicia faba MT genes that belong to the type 1 group of plant MT genes in contrast to a MT gene we previously isolated that belongs to type 2. We found similarities and differences between the V. faba MT genes. The RNA expression patterns differed and this was most pronounced in roots, which contained high MT1 but very low MT2 RNA levels. Like MT2, MT1 transcript levels were not significantly affected by treatment with Cd, Cu, Fe and Zn, at least under the experimental conditions. MT RNA levels varied in leaves and stem internodes of different developmental ages, with the highest expression in the older tissue. The levels of MT RNA correlated inversely with endogenous Cd, Cu and Fe levels within different internodes, but not with a number of other metals tested (including Zn). The three bean MTs were expressed in Escherichia coli and found to bind Cd, Cu and Zn but not to Fe. The MTs were tested to determine if they differed in their ability to bind a specific metal but no significant differences in binding were observed.

Plant Molecular Biology, 1994

Animal metallothioneins (MTs) are cysteine-rich, low-molecular-weight proteins that bind to heavy... more Animal metallothioneins (MTs) are cysteine-rich, low-molecular-weight proteins that bind to heavy metals and are believed to play a role in their metabolism and detoxification. Genes encoding MT-like proteins have been isolated in a number of plants although their function remains to be elucidated. We describe the isolation and characterization of a bean cDNA encoding an MT-like protein. The bean gene, called MT, was isolated as a result of a differential screen for genes that are expressed in leaves but not in the most common cell type, the mesophyll cell. MT contained two regions with abundant cysteines and sequence comparison found that MT had greatest homology to MT-like subtype 2 from other plant species. Northern blot analysis demonstrated that MT was expressed in the leaf, stem and flower, at very low levels in roots and was not detectable in mesophyll protoplasts. MT transcript levels were not significantly affected by treatment with Cu, Zn or Cd. In the leaf, in situ hybridization studies demonstrated striking cell specificity with MT expression confined predominantly to trichomes. Possible explanations for the pronounced expression of MT in leaf trichomes are discussed.

Molecular Plant-microbe Interactions, 2007

Aphids are major insect pests of plants that feed directly from the phloem. We used the model leg... more Aphids are major insect pests of plants that feed directly from the phloem. We used the model legume Medicago truncatula Gaert. (barrel medic) to elucidate host resistance to aphids and identified a single dominant gene which confers resistance to Acyrthosiphon kondoi Shinji (bluegreen aphid). To understand how this gene conditions resistance to bluegreen aphid, transcription profiling of 23 defense-related genes representing various signaling pathways was undertaken using a pair of near-isogenic lines that are susceptible or resistant to bluegreen aphid. All salicylic acid- and ethylene-responsive genes tested were induced by bluegreen aphid in resistant and susceptible plants, although there were some differences in the magnitude and kinetics of the induction. In contrast, 10 of 13 genes associated with the octadecanoid pathway were induced exclusively in the resistant plants following bluegreen aphid infestation. These results are in contrast to plant-pathogen interactions where similar sets of defense genes typically are induced in compatible interactions, but to a lesser degree and later than in incompatible interactions. Treatment of susceptible plants with methyl jasmonate reduced bluegreen aphid infestation but not to the same levels as the resistant line. Together, these results strongly suggest that the octadecanoid pathway is important for this naturally derived aphid resistance trait.

Plant Journal, 1993

Ocs-elements are a group of related, bipartite promoter elements which have been exploited by two... more Ocs-elements are a group of related, bipartite promoter elements which have been exploited by two distinct groups of plant pathogens, Agrobacterium and certain viruses to express genes in plants. The genes for two Arabidopsis bZIP (basic region-leucine zipper) proteins that bind to ocs-elements have been isolated and characterized. The genes, called OBF4 and OBF5, were isolated by screening an Arabidopsis genomic library with degenerate oligonucleotides complementary to the DNA-binding domains of other plant ocs-element-binding proteins. The OBF4 and OBF5 proteins show 53% amino acid identity but low DNA homology. Southern blot analysis demonstrated that each of the OBF genes is a member of a small family. OBF4 is more similar to the tobacco TGA1a and Arabidopsis TGA1 proteins, while OBF5 is more similar to the maize OBF3.1, wheat HBP1b and Arabidopsis aHBP1b proteins. The DNA-binding properties of OBF4 and OBF5 were similar although OBF5 was able to bind simultaneously to both halves of the ocs-element more efficiently than OBF4. This difference in binding to the ocs-element between two closely related proteins from the same species is potentially significant since binding to both halves of the ocs-element is a pre-requisite for in vivo transcriptional activity.

Plant Journal, 2003

Overexpression of a salicylic-acid (SA)-inducible Arabidopsis DNA binding with one finger (Dof) t... more Overexpression of a salicylic-acid (SA)-inducible Arabidopsis DNA binding with one finger (Dof) transcription factor, called OBF-binding protein 3 (OBP3; AtDof3.6), has previously been shown to result in growth defects. In this study, suppressive subtraction hybridization (SSH) was used to isolate genes induced in an OBP3-overexpression line and several putative clones, called OBP3-responsive genes (ORGs), were isolated. The link with the induced expression levels of these genes and OBP3 overexpression was confirmed by analysing additional OBP3-overexpression lines. ORG1 through ORG4 are novel genes, while ORG5 is an extensin gene, AtExt1. While ORG4 has no similarity with other proteins in the database, ORG1 has weak similarity in different regions of the predicted protein with CDC2 and fibrillin. ORG2 and ORG3 share 80% overall identity in their deduced amino acid sequences and contain a basic helix-loop-helix DNA-binding domain, suggesting that ORG2 and ORG3 may be transcription factors. The expression of the ORG1, ORG2 and ORG3 genes was co-regulated under all conditions examined including upregulation by SA and downregulation by jasmonic acid (JA). Fifteen OBP3-silenced lines were generated to further explore the function of OBP3. Although there were no visible phenotypic changes in any of these lines, the expression of ORG1, ORG2 and ORG3 was reduced. Among the ORG genes, ORG1, ORG2 and ORG3 contained the highest number of potential Dof-binding sites in the promoter region, and their expression was significantly increased within 3 h after induction of OBP3 expression using an inducible promoter system, and closely reflected the expression levels of the exogenous OBP3 protein. The results from the gain-of-function and loss-of-function experiments suggest that the ORG1, ORG2 and ORG3 genes are direct target genes of OBP3.