The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Jiahua Xie - One of the best experts on this subject based on the ideXlab platform.
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transformation of long lived albino Epipremnum aureum golden pothos and restoring chloroplast development
Frontiers in Plant Science, 2021Co-Authors: Chiu-yueh Hung, Farooqahmed S. Kittur, Jianjun Chen, Kent O Burkey, Jianhui Zhang, Chayanika Bhattacharya, Carla E Oldham, Xiangying Wei, Jiahua XieAbstract:Chloroplasts are organelles responsible for chlorophyll biosynthesis and photosynthesis, which are one of key targets for crop improvement. Elucidating and engineering genes involved in chlorophyll biosynthesis are important approaches for studying chloroplast development and biogenesis as well as developing new crops. In this study, we report a long-lived albino mutant derived from a popular ornamental plant Epipremnum aureum ‘Golden Pothos’ which could be used as a model for analyzing the function of genes involved in chlorophyll biosynthesis and generating colorful plants. Albino mutant plants were isolated from regenerated populations of variegated ‘Golden Pothos’ whose albino phenotype was previously found to be due to impaired expression of EaZIP, encoding Mg-protoporphyrin IX monomethyl ester cyclase. The mutant plants have been maintained in MS medium for 11 years. Using petioles of the mutant plants as explants with a traceable sGFP gene, an efficient transformation system was developed. Overexpressing Arabidopsis CHL27 (a homolog of EaZIP) but not EaZIP in albino plants restored green color and chloroplast development. Interestingly, in addition to the occurrence of plants with solid green color, plants with variegated leaves and pale-yellow leaves were also obtained in the regenerated populations, suggesting that green color restoration might be resulted from the interaction between the gene and environmental factor(s). Nevertheless, our study shows that these long-lived albino plants along with the established efficient transformation system could be used for creating colorful ornamental plants. This system could potentially be used for investigating physiological processes associated with chlorophyll biosynthesis and chloroplast development as well as various biological activities, which are difficult to achieve using green plants.
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differential expression of a novel gene eaf82a in green and yellow sectors of variegated Epipremnum aureum leaves is related to uneven distribution of auxin
Physiologia Plantarum, 2014Co-Authors: Chiu-yueh Hung, Farooqahmed S. Kittur, Richard J Henny, Jianjun Chen, Kent O Burkey, Makendra Umstead, Bronwyn M Holliday, Jiahua XieAbstract:EaF82, a gene identified in previous studies of the variegated plant Epipremnum aureum, exhibited a unique expression pattern with greater transcript abundance in yellow sectors than green sectors of variegated leaves, but lower abundance in regenerated pale yellow plants than in green plants derived from leaf tissue culture. Studies of its full-length cDNA and promoter region revealed two members with only the EaF82a expressed. Immunoblotting confirmed that EaF82a encodes a 12 kDa protein and its accumulation consistent with its gene expression patterns in different color tissues. Transient expression of EaF82a-sGFP fusion proteins in protoplasts showed that EaF82a seems to be present in the cytosol as unidentified spots. Sequence motif search reveals a potential auxin responsive element in promoter region. Using transgenic Arabidopsis seedlings carrying EaF82a promoter driving the bacterial uidA (GUS) gene, an increased GUS activity was observed when IAA (indole-3-acetic acid) concentration was elevated. In E. aureum, EaF82a is more abundant at the site where axillary buds emerge and at the lower side of bending nodes where more IAA accumulates relative to the upper side. The measurement of endogenous IAA levels in different color tissues revealed the same pattern of IAA distribution as that of EaF82a expression, further supporting that EaF82a is an IAA responsive gene. EaF82a expression in etiolated transgenic Arabidopsis seedlings responded to IAA under the influence of light suggesting a microenvironment of uneven light condition affects the EaF82a transcript levels and protein accumulation in variegated leaves.
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efficient somatic embryogenesis and agrobacterium mediated transformation of pothos Epipremnum aureum jade
Plant Cell Tissue and Organ Culture, 2013Co-Authors: Jietang Zhao, Richard J Henny, Jiahua Xie, Jin Cui, Dennis J Gray, Jianjun ChenAbstract:Leaf and petiole explants of monocotyledonous pothos (Epipremnum aureum) ‘Jade’ were cultured on Murashige and Skoog basal medium supplemented with N-(2-chloro-4-pyridl)-N′-phenylurea (CPPU) or N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ) with α-naphthalene acetic acid (NAA). Somatic embryos appeared directly from explants after 4–8 weeks of culture; 9.1 μM TDZ with 1.1 μM NAA induced 61.1 % leaf discs and 94.4 % of petiole segments to produce plantlets through embryo conversion. Using this established regeneration method and an enhanced green fluorescent protein (GFP) gene (egfp) as a reporter marker, an Agrobacterium-mediated transformation procedure was developed. Leaf discs and petiole segments were inoculated with Agrobacterium tumefaciens strain EHA105 harboring a binary vector pLC902 that contains novel bi-directional duplex promoters driving the egfp gene and hygromycin phosphotransferase gene (hpt), respectively. The explants were co-cultivated with strain EHA105 for 3, 5, and 7 days, respectively prior to selective culture with 25 mg l−1 hygromycin. A 5-day co-cultivation led to 100 % of leaf discs to show transient GFP expression and 23.8 % of the discs to produce stable GFP-expressing somatic embryos. A 7-day co-cultivation of petiole explants resulted in the corresponding responses at 100 and 14.3 %, respectively. A total of 237 transgenic plants were obtained, and GFP fluorescence was observed in all plant organs. Regular PCR and quantitative real-time PCR analyses confirmed the presence of 1 or 2 copies of the egfp gene in analyzed plants. The highly efficient regeneration and transformation systems established in this study may enable genetic improvement of this vegetatively propagated species through biotechnological means.
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plant regeneration via direct somatic embryogenesis from leaf and petiole explants of Epipremnum aureum marble queen and characterization of selected variants
Acta Physiologiae Plantarum, 2012Co-Authors: Jietang Zhao, Chiu-yueh Hung, Richard J Henny, Jiahua Xie, Jin Cui, Qian Zhang, Jianjun ChenAbstract:Leaf and petiole explants of Epipremnum aureum ‘Marble Queen’ were cultured on Murashige and Skoog basal medium containing three concentrations of either N-(2-chloro-4-pyridl)-N’-phenylurea (CPPU) or N-phenyl-N’-1, 2, 3-thiadiazol-5-ylurea (TDZ) with 1.07 μM α-naphthalene acetic acid (NAA). Somatic embryos appeared directly from explants after 4–6 weeks of culture. TDZ at 4.54 μM with 1.07 μM NAA induced 75% of leaf explants to produce somatic embryos and 62.5% of explants to produce germinated embryos. Both 8.07 μM CPPU and 9.08 μM TDZ, respectively, with 1.07 μM NAA induced 100% of petiole explants to produce somatic embryos and 78.1 and 91.7% of explants to produce germinated embryos. Plantlets with completely green, variegated, and whitish leaves were identified among regenerated individuals. Some variegation patterns differed greatly from the parent ‘Marble Queen’ within the variegated plants. Flow cytometry analysis of stock plants and selected variants showed that all plants had one identical peak. Analysis of simple sequence repeats amplified from 14 universal chloroplast primer pairs showed no variation between stock plants and selected variants. RT-PCR analysis of EaZIP, a marker gene involved in leaf variegation of E. aureum ‘Golden Pothos’, suggested little expression difference between green and whitish plants or between green and whitish sectors of a variegated leaf. However, SDS-PAGE analysis of proteins showed differences in band intensity and patterns. These results suggest that mechanisms underlying leaf variegation of ‘Marble Queen’ differ from the closely related ‘Golden Pothos’. The established regeneration system and identified variants could be important materials for further investigation of leaf variegation in E. aureum.
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identification of a mg protoporphyrin ix monomethyl ester cyclase homologue eazip differentially expressed in variegated Epipremnum aureum golden pothos is achieved through a unique method of comparative study using tissue regenerated plants
Journal of Experimental Botany, 2010Co-Authors: Chiu-yueh Hung, Jianjun Chen, Yinghsuan Sun, Diane E Darlington, Alfred L Williams, Kent O Burkey, Jiahua XieAbstract:Variegated plants provide a valuable tool for studying chloroplast biogenesis by allowing direct comparison between green and white/yellow sectors within the same leaf. While variegated plants are abundant in nature, the mechanism of leaf variegation remains largely unknown. Current studies are limited to a few mutants in model plant species, and are complicated by the potential for cross-contamination during dissection of leaf tissue into contrasting sectors. To overcome these obstacles, an alternative approach was explored using tissue-culture techniques to regenerate plantlets from unique sectors. Stable green and pale yellow plants were developed from a naturally variegated Epipremnum aureum ‘Golden Pothos’. By comparing the gene expression between green and pale yellow plants using suppression subtractive hybridization in conjunction with homologous sequence search, nine down-regulated and 18 up-regulated genes were identified in pale yellow plants. Transcript abundance for EaZIP (Epipremnum aureum leucine zipper), a nuclear gene homologue of tobacco NTZIP and Arabidopsis CHL27, was reduced more than 4000-fold in qRT-PCR analysis. EaZIP encodes the Mg-protoporphyrin IX monomethyl ester cyclase, one of the key enzymes in the chlorophyll biosynthesis pathway. Examination of EaZIP expression in naturally variegated ‘Golden Pothos’ confirmed that EaZIP transcript levels were correlated with leaf chlorophyll contents, suggesting that this gene plays a major role in the loss of chlorophyll in the pale yellow sectors of E. aureum ‘Golden Pothos’. This study further suggests that tissue-culture regeneration of plantlets from different coloured sectors of variegated leaves can be used to investigate the underlying mechanisms of variegation.
Jianjun Chen - One of the best experts on this subject based on the ideXlab platform.
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transformation of long lived albino Epipremnum aureum golden pothos and restoring chloroplast development
Frontiers in Plant Science, 2021Co-Authors: Chiu-yueh Hung, Farooqahmed S. Kittur, Jianjun Chen, Kent O Burkey, Jianhui Zhang, Chayanika Bhattacharya, Carla E Oldham, Xiangying Wei, Jiahua XieAbstract:Chloroplasts are organelles responsible for chlorophyll biosynthesis and photosynthesis, which are one of key targets for crop improvement. Elucidating and engineering genes involved in chlorophyll biosynthesis are important approaches for studying chloroplast development and biogenesis as well as developing new crops. In this study, we report a long-lived albino mutant derived from a popular ornamental plant Epipremnum aureum ‘Golden Pothos’ which could be used as a model for analyzing the function of genes involved in chlorophyll biosynthesis and generating colorful plants. Albino mutant plants were isolated from regenerated populations of variegated ‘Golden Pothos’ whose albino phenotype was previously found to be due to impaired expression of EaZIP, encoding Mg-protoporphyrin IX monomethyl ester cyclase. The mutant plants have been maintained in MS medium for 11 years. Using petioles of the mutant plants as explants with a traceable sGFP gene, an efficient transformation system was developed. Overexpressing Arabidopsis CHL27 (a homolog of EaZIP) but not EaZIP in albino plants restored green color and chloroplast development. Interestingly, in addition to the occurrence of plants with solid green color, plants with variegated leaves and pale-yellow leaves were also obtained in the regenerated populations, suggesting that green color restoration might be resulted from the interaction between the gene and environmental factor(s). Nevertheless, our study shows that these long-lived albino plants along with the established efficient transformation system could be used for creating colorful ornamental plants. This system could potentially be used for investigating physiological processes associated with chlorophyll biosynthesis and chloroplast development as well as various biological activities, which are difficult to achieve using green plants.
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Gibberellin deficiency is responsible for shy-flowering nature of Epipremnum aureum.
Scientific Reports, 2016Co-Authors: Chiu-yueh Hung, Farooqahmed S. Kittur, Jianjun Chen, Richard J HennyAbstract:Epipremnum aureum is an extremely popular houseplant belonging to the Araceae family of angiosperms, but it does not flower either in the wild or under cultivation. We uncovered the potential causes of its shy-flowering nature by building the transcriptome using next-generation sequencing and identifying floral-related genes that are differentially expressed between vertical growth (VG, adult) and horizontal growth (HG, juvenile) plants. Transcripts of the gibberellin (GA) biosynthetic gene EaGA3ox1 and GA-responsive floral meristem identity gene EaLFY were absent in both VG and HG plants, suggesting that a deficiency of bioactive GAs may be responsible for its shy-flowering nature. This hypothesis is supported by undetectable or low levels of three bioactive GAs, and exogenous GA3 triggered flowering in both plants. Our study resolves the mystery why E. aureum fails to flower, and reveals the positive role of GAs in floral transition in perennials.
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differential expression of a novel gene eaf82a in green and yellow sectors of variegated Epipremnum aureum leaves is related to uneven distribution of auxin
Physiologia Plantarum, 2014Co-Authors: Chiu-yueh Hung, Farooqahmed S. Kittur, Richard J Henny, Jianjun Chen, Kent O Burkey, Makendra Umstead, Bronwyn M Holliday, Jiahua XieAbstract:EaF82, a gene identified in previous studies of the variegated plant Epipremnum aureum, exhibited a unique expression pattern with greater transcript abundance in yellow sectors than green sectors of variegated leaves, but lower abundance in regenerated pale yellow plants than in green plants derived from leaf tissue culture. Studies of its full-length cDNA and promoter region revealed two members with only the EaF82a expressed. Immunoblotting confirmed that EaF82a encodes a 12 kDa protein and its accumulation consistent with its gene expression patterns in different color tissues. Transient expression of EaF82a-sGFP fusion proteins in protoplasts showed that EaF82a seems to be present in the cytosol as unidentified spots. Sequence motif search reveals a potential auxin responsive element in promoter region. Using transgenic Arabidopsis seedlings carrying EaF82a promoter driving the bacterial uidA (GUS) gene, an increased GUS activity was observed when IAA (indole-3-acetic acid) concentration was elevated. In E. aureum, EaF82a is more abundant at the site where axillary buds emerge and at the lower side of bending nodes where more IAA accumulates relative to the upper side. The measurement of endogenous IAA levels in different color tissues revealed the same pattern of IAA distribution as that of EaF82a expression, further supporting that EaF82a is an IAA responsive gene. EaF82a expression in etiolated transgenic Arabidopsis seedlings responded to IAA under the influence of light suggesting a microenvironment of uneven light condition affects the EaF82a transcript levels and protein accumulation in variegated leaves.
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efficient somatic embryogenesis and agrobacterium mediated transformation of pothos Epipremnum aureum jade
Plant Cell Tissue and Organ Culture, 2013Co-Authors: Jietang Zhao, Richard J Henny, Jiahua Xie, Jin Cui, Dennis J Gray, Jianjun ChenAbstract:Leaf and petiole explants of monocotyledonous pothos (Epipremnum aureum) ‘Jade’ were cultured on Murashige and Skoog basal medium supplemented with N-(2-chloro-4-pyridl)-N′-phenylurea (CPPU) or N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ) with α-naphthalene acetic acid (NAA). Somatic embryos appeared directly from explants after 4–8 weeks of culture; 9.1 μM TDZ with 1.1 μM NAA induced 61.1 % leaf discs and 94.4 % of petiole segments to produce plantlets through embryo conversion. Using this established regeneration method and an enhanced green fluorescent protein (GFP) gene (egfp) as a reporter marker, an Agrobacterium-mediated transformation procedure was developed. Leaf discs and petiole segments were inoculated with Agrobacterium tumefaciens strain EHA105 harboring a binary vector pLC902 that contains novel bi-directional duplex promoters driving the egfp gene and hygromycin phosphotransferase gene (hpt), respectively. The explants were co-cultivated with strain EHA105 for 3, 5, and 7 days, respectively prior to selective culture with 25 mg l−1 hygromycin. A 5-day co-cultivation led to 100 % of leaf discs to show transient GFP expression and 23.8 % of the discs to produce stable GFP-expressing somatic embryos. A 7-day co-cultivation of petiole explants resulted in the corresponding responses at 100 and 14.3 %, respectively. A total of 237 transgenic plants were obtained, and GFP fluorescence was observed in all plant organs. Regular PCR and quantitative real-time PCR analyses confirmed the presence of 1 or 2 copies of the egfp gene in analyzed plants. The highly efficient regeneration and transformation systems established in this study may enable genetic improvement of this vegetatively propagated species through biotechnological means.
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plant regeneration via direct somatic embryogenesis from leaf and petiole explants of Epipremnum aureum marble queen and characterization of selected variants
Acta Physiologiae Plantarum, 2012Co-Authors: Jietang Zhao, Chiu-yueh Hung, Richard J Henny, Jiahua Xie, Jin Cui, Qian Zhang, Jianjun ChenAbstract:Leaf and petiole explants of Epipremnum aureum ‘Marble Queen’ were cultured on Murashige and Skoog basal medium containing three concentrations of either N-(2-chloro-4-pyridl)-N’-phenylurea (CPPU) or N-phenyl-N’-1, 2, 3-thiadiazol-5-ylurea (TDZ) with 1.07 μM α-naphthalene acetic acid (NAA). Somatic embryos appeared directly from explants after 4–6 weeks of culture. TDZ at 4.54 μM with 1.07 μM NAA induced 75% of leaf explants to produce somatic embryos and 62.5% of explants to produce germinated embryos. Both 8.07 μM CPPU and 9.08 μM TDZ, respectively, with 1.07 μM NAA induced 100% of petiole explants to produce somatic embryos and 78.1 and 91.7% of explants to produce germinated embryos. Plantlets with completely green, variegated, and whitish leaves were identified among regenerated individuals. Some variegation patterns differed greatly from the parent ‘Marble Queen’ within the variegated plants. Flow cytometry analysis of stock plants and selected variants showed that all plants had one identical peak. Analysis of simple sequence repeats amplified from 14 universal chloroplast primer pairs showed no variation between stock plants and selected variants. RT-PCR analysis of EaZIP, a marker gene involved in leaf variegation of E. aureum ‘Golden Pothos’, suggested little expression difference between green and whitish plants or between green and whitish sectors of a variegated leaf. However, SDS-PAGE analysis of proteins showed differences in band intensity and patterns. These results suggest that mechanisms underlying leaf variegation of ‘Marble Queen’ differ from the closely related ‘Golden Pothos’. The established regeneration system and identified variants could be important materials for further investigation of leaf variegation in E. aureum.
Chiu-yueh Hung - One of the best experts on this subject based on the ideXlab platform.
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transformation of long lived albino Epipremnum aureum golden pothos and restoring chloroplast development
Frontiers in Plant Science, 2021Co-Authors: Chiu-yueh Hung, Farooqahmed S. Kittur, Jianjun Chen, Kent O Burkey, Jianhui Zhang, Chayanika Bhattacharya, Carla E Oldham, Xiangying Wei, Jiahua XieAbstract:Chloroplasts are organelles responsible for chlorophyll biosynthesis and photosynthesis, which are one of key targets for crop improvement. Elucidating and engineering genes involved in chlorophyll biosynthesis are important approaches for studying chloroplast development and biogenesis as well as developing new crops. In this study, we report a long-lived albino mutant derived from a popular ornamental plant Epipremnum aureum ‘Golden Pothos’ which could be used as a model for analyzing the function of genes involved in chlorophyll biosynthesis and generating colorful plants. Albino mutant plants were isolated from regenerated populations of variegated ‘Golden Pothos’ whose albino phenotype was previously found to be due to impaired expression of EaZIP, encoding Mg-protoporphyrin IX monomethyl ester cyclase. The mutant plants have been maintained in MS medium for 11 years. Using petioles of the mutant plants as explants with a traceable sGFP gene, an efficient transformation system was developed. Overexpressing Arabidopsis CHL27 (a homolog of EaZIP) but not EaZIP in albino plants restored green color and chloroplast development. Interestingly, in addition to the occurrence of plants with solid green color, plants with variegated leaves and pale-yellow leaves were also obtained in the regenerated populations, suggesting that green color restoration might be resulted from the interaction between the gene and environmental factor(s). Nevertheless, our study shows that these long-lived albino plants along with the established efficient transformation system could be used for creating colorful ornamental plants. This system could potentially be used for investigating physiological processes associated with chlorophyll biosynthesis and chloroplast development as well as various biological activities, which are difficult to achieve using green plants.
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Gibberellin deficiency is responsible for shy-flowering nature of Epipremnum aureum.
Scientific Reports, 2016Co-Authors: Chiu-yueh Hung, Farooqahmed S. Kittur, Jianjun Chen, Richard J HennyAbstract:Epipremnum aureum is an extremely popular houseplant belonging to the Araceae family of angiosperms, but it does not flower either in the wild or under cultivation. We uncovered the potential causes of its shy-flowering nature by building the transcriptome using next-generation sequencing and identifying floral-related genes that are differentially expressed between vertical growth (VG, adult) and horizontal growth (HG, juvenile) plants. Transcripts of the gibberellin (GA) biosynthetic gene EaGA3ox1 and GA-responsive floral meristem identity gene EaLFY were absent in both VG and HG plants, suggesting that a deficiency of bioactive GAs may be responsible for its shy-flowering nature. This hypothesis is supported by undetectable or low levels of three bioactive GAs, and exogenous GA3 triggered flowering in both plants. Our study resolves the mystery why E. aureum fails to flower, and reveals the positive role of GAs in floral transition in perennials.
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differential expression of a novel gene eaf82a in green and yellow sectors of variegated Epipremnum aureum leaves is related to uneven distribution of auxin
Physiologia Plantarum, 2014Co-Authors: Chiu-yueh Hung, Farooqahmed S. Kittur, Richard J Henny, Jianjun Chen, Kent O Burkey, Makendra Umstead, Bronwyn M Holliday, Jiahua XieAbstract:EaF82, a gene identified in previous studies of the variegated plant Epipremnum aureum, exhibited a unique expression pattern with greater transcript abundance in yellow sectors than green sectors of variegated leaves, but lower abundance in regenerated pale yellow plants than in green plants derived from leaf tissue culture. Studies of its full-length cDNA and promoter region revealed two members with only the EaF82a expressed. Immunoblotting confirmed that EaF82a encodes a 12 kDa protein and its accumulation consistent with its gene expression patterns in different color tissues. Transient expression of EaF82a-sGFP fusion proteins in protoplasts showed that EaF82a seems to be present in the cytosol as unidentified spots. Sequence motif search reveals a potential auxin responsive element in promoter region. Using transgenic Arabidopsis seedlings carrying EaF82a promoter driving the bacterial uidA (GUS) gene, an increased GUS activity was observed when IAA (indole-3-acetic acid) concentration was elevated. In E. aureum, EaF82a is more abundant at the site where axillary buds emerge and at the lower side of bending nodes where more IAA accumulates relative to the upper side. The measurement of endogenous IAA levels in different color tissues revealed the same pattern of IAA distribution as that of EaF82a expression, further supporting that EaF82a is an IAA responsive gene. EaF82a expression in etiolated transgenic Arabidopsis seedlings responded to IAA under the influence of light suggesting a microenvironment of uneven light condition affects the EaF82a transcript levels and protein accumulation in variegated leaves.
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plant regeneration via direct somatic embryogenesis from leaf and petiole explants of Epipremnum aureum marble queen and characterization of selected variants
Acta Physiologiae Plantarum, 2012Co-Authors: Jietang Zhao, Chiu-yueh Hung, Richard J Henny, Jiahua Xie, Jin Cui, Qian Zhang, Jianjun ChenAbstract:Leaf and petiole explants of Epipremnum aureum ‘Marble Queen’ were cultured on Murashige and Skoog basal medium containing three concentrations of either N-(2-chloro-4-pyridl)-N’-phenylurea (CPPU) or N-phenyl-N’-1, 2, 3-thiadiazol-5-ylurea (TDZ) with 1.07 μM α-naphthalene acetic acid (NAA). Somatic embryos appeared directly from explants after 4–6 weeks of culture. TDZ at 4.54 μM with 1.07 μM NAA induced 75% of leaf explants to produce somatic embryos and 62.5% of explants to produce germinated embryos. Both 8.07 μM CPPU and 9.08 μM TDZ, respectively, with 1.07 μM NAA induced 100% of petiole explants to produce somatic embryos and 78.1 and 91.7% of explants to produce germinated embryos. Plantlets with completely green, variegated, and whitish leaves were identified among regenerated individuals. Some variegation patterns differed greatly from the parent ‘Marble Queen’ within the variegated plants. Flow cytometry analysis of stock plants and selected variants showed that all plants had one identical peak. Analysis of simple sequence repeats amplified from 14 universal chloroplast primer pairs showed no variation between stock plants and selected variants. RT-PCR analysis of EaZIP, a marker gene involved in leaf variegation of E. aureum ‘Golden Pothos’, suggested little expression difference between green and whitish plants or between green and whitish sectors of a variegated leaf. However, SDS-PAGE analysis of proteins showed differences in band intensity and patterns. These results suggest that mechanisms underlying leaf variegation of ‘Marble Queen’ differ from the closely related ‘Golden Pothos’. The established regeneration system and identified variants could be important materials for further investigation of leaf variegation in E. aureum.
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identification of a mg protoporphyrin ix monomethyl ester cyclase homologue eazip differentially expressed in variegated Epipremnum aureum golden pothos is achieved through a unique method of comparative study using tissue regenerated plants
Journal of Experimental Botany, 2010Co-Authors: Chiu-yueh Hung, Jianjun Chen, Yinghsuan Sun, Diane E Darlington, Alfred L Williams, Kent O Burkey, Jiahua XieAbstract:Variegated plants provide a valuable tool for studying chloroplast biogenesis by allowing direct comparison between green and white/yellow sectors within the same leaf. While variegated plants are abundant in nature, the mechanism of leaf variegation remains largely unknown. Current studies are limited to a few mutants in model plant species, and are complicated by the potential for cross-contamination during dissection of leaf tissue into contrasting sectors. To overcome these obstacles, an alternative approach was explored using tissue-culture techniques to regenerate plantlets from unique sectors. Stable green and pale yellow plants were developed from a naturally variegated Epipremnum aureum ‘Golden Pothos’. By comparing the gene expression between green and pale yellow plants using suppression subtractive hybridization in conjunction with homologous sequence search, nine down-regulated and 18 up-regulated genes were identified in pale yellow plants. Transcript abundance for EaZIP (Epipremnum aureum leucine zipper), a nuclear gene homologue of tobacco NTZIP and Arabidopsis CHL27, was reduced more than 4000-fold in qRT-PCR analysis. EaZIP encodes the Mg-protoporphyrin IX monomethyl ester cyclase, one of the key enzymes in the chlorophyll biosynthesis pathway. Examination of EaZIP expression in naturally variegated ‘Golden Pothos’ confirmed that EaZIP transcript levels were correlated with leaf chlorophyll contents, suggesting that this gene plays a major role in the loss of chlorophyll in the pale yellow sectors of E. aureum ‘Golden Pothos’. This study further suggests that tissue-culture regeneration of plantlets from different coloured sectors of variegated leaves can be used to investigate the underlying mechanisms of variegation.
Richard J Henny - One of the best experts on this subject based on the ideXlab platform.
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Gibberellin deficiency is responsible for shy-flowering nature of Epipremnum aureum.
Scientific Reports, 2016Co-Authors: Chiu-yueh Hung, Farooqahmed S. Kittur, Jianjun Chen, Richard J HennyAbstract:Epipremnum aureum is an extremely popular houseplant belonging to the Araceae family of angiosperms, but it does not flower either in the wild or under cultivation. We uncovered the potential causes of its shy-flowering nature by building the transcriptome using next-generation sequencing and identifying floral-related genes that are differentially expressed between vertical growth (VG, adult) and horizontal growth (HG, juvenile) plants. Transcripts of the gibberellin (GA) biosynthetic gene EaGA3ox1 and GA-responsive floral meristem identity gene EaLFY were absent in both VG and HG plants, suggesting that a deficiency of bioactive GAs may be responsible for its shy-flowering nature. This hypothesis is supported by undetectable or low levels of three bioactive GAs, and exogenous GA3 triggered flowering in both plants. Our study resolves the mystery why E. aureum fails to flower, and reveals the positive role of GAs in floral transition in perennials.
-
differential expression of a novel gene eaf82a in green and yellow sectors of variegated Epipremnum aureum leaves is related to uneven distribution of auxin
Physiologia Plantarum, 2014Co-Authors: Chiu-yueh Hung, Farooqahmed S. Kittur, Richard J Henny, Jianjun Chen, Kent O Burkey, Makendra Umstead, Bronwyn M Holliday, Jiahua XieAbstract:EaF82, a gene identified in previous studies of the variegated plant Epipremnum aureum, exhibited a unique expression pattern with greater transcript abundance in yellow sectors than green sectors of variegated leaves, but lower abundance in regenerated pale yellow plants than in green plants derived from leaf tissue culture. Studies of its full-length cDNA and promoter region revealed two members with only the EaF82a expressed. Immunoblotting confirmed that EaF82a encodes a 12 kDa protein and its accumulation consistent with its gene expression patterns in different color tissues. Transient expression of EaF82a-sGFP fusion proteins in protoplasts showed that EaF82a seems to be present in the cytosol as unidentified spots. Sequence motif search reveals a potential auxin responsive element in promoter region. Using transgenic Arabidopsis seedlings carrying EaF82a promoter driving the bacterial uidA (GUS) gene, an increased GUS activity was observed when IAA (indole-3-acetic acid) concentration was elevated. In E. aureum, EaF82a is more abundant at the site where axillary buds emerge and at the lower side of bending nodes where more IAA accumulates relative to the upper side. The measurement of endogenous IAA levels in different color tissues revealed the same pattern of IAA distribution as that of EaF82a expression, further supporting that EaF82a is an IAA responsive gene. EaF82a expression in etiolated transgenic Arabidopsis seedlings responded to IAA under the influence of light suggesting a microenvironment of uneven light condition affects the EaF82a transcript levels and protein accumulation in variegated leaves.
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efficient somatic embryogenesis and agrobacterium mediated transformation of pothos Epipremnum aureum jade
Plant Cell Tissue and Organ Culture, 2013Co-Authors: Jietang Zhao, Richard J Henny, Jiahua Xie, Jin Cui, Dennis J Gray, Jianjun ChenAbstract:Leaf and petiole explants of monocotyledonous pothos (Epipremnum aureum) ‘Jade’ were cultured on Murashige and Skoog basal medium supplemented with N-(2-chloro-4-pyridl)-N′-phenylurea (CPPU) or N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ) with α-naphthalene acetic acid (NAA). Somatic embryos appeared directly from explants after 4–8 weeks of culture; 9.1 μM TDZ with 1.1 μM NAA induced 61.1 % leaf discs and 94.4 % of petiole segments to produce plantlets through embryo conversion. Using this established regeneration method and an enhanced green fluorescent protein (GFP) gene (egfp) as a reporter marker, an Agrobacterium-mediated transformation procedure was developed. Leaf discs and petiole segments were inoculated with Agrobacterium tumefaciens strain EHA105 harboring a binary vector pLC902 that contains novel bi-directional duplex promoters driving the egfp gene and hygromycin phosphotransferase gene (hpt), respectively. The explants were co-cultivated with strain EHA105 for 3, 5, and 7 days, respectively prior to selective culture with 25 mg l−1 hygromycin. A 5-day co-cultivation led to 100 % of leaf discs to show transient GFP expression and 23.8 % of the discs to produce stable GFP-expressing somatic embryos. A 7-day co-cultivation of petiole explants resulted in the corresponding responses at 100 and 14.3 %, respectively. A total of 237 transgenic plants were obtained, and GFP fluorescence was observed in all plant organs. Regular PCR and quantitative real-time PCR analyses confirmed the presence of 1 or 2 copies of the egfp gene in analyzed plants. The highly efficient regeneration and transformation systems established in this study may enable genetic improvement of this vegetatively propagated species through biotechnological means.
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plant regeneration via direct somatic embryogenesis from leaf and petiole explants of Epipremnum aureum marble queen and characterization of selected variants
Acta Physiologiae Plantarum, 2012Co-Authors: Jietang Zhao, Chiu-yueh Hung, Richard J Henny, Jiahua Xie, Jin Cui, Qian Zhang, Jianjun ChenAbstract:Leaf and petiole explants of Epipremnum aureum ‘Marble Queen’ were cultured on Murashige and Skoog basal medium containing three concentrations of either N-(2-chloro-4-pyridl)-N’-phenylurea (CPPU) or N-phenyl-N’-1, 2, 3-thiadiazol-5-ylurea (TDZ) with 1.07 μM α-naphthalene acetic acid (NAA). Somatic embryos appeared directly from explants after 4–6 weeks of culture. TDZ at 4.54 μM with 1.07 μM NAA induced 75% of leaf explants to produce somatic embryos and 62.5% of explants to produce germinated embryos. Both 8.07 μM CPPU and 9.08 μM TDZ, respectively, with 1.07 μM NAA induced 100% of petiole explants to produce somatic embryos and 78.1 and 91.7% of explants to produce germinated embryos. Plantlets with completely green, variegated, and whitish leaves were identified among regenerated individuals. Some variegation patterns differed greatly from the parent ‘Marble Queen’ within the variegated plants. Flow cytometry analysis of stock plants and selected variants showed that all plants had one identical peak. Analysis of simple sequence repeats amplified from 14 universal chloroplast primer pairs showed no variation between stock plants and selected variants. RT-PCR analysis of EaZIP, a marker gene involved in leaf variegation of E. aureum ‘Golden Pothos’, suggested little expression difference between green and whitish plants or between green and whitish sectors of a variegated leaf. However, SDS-PAGE analysis of proteins showed differences in band intensity and patterns. These results suggest that mechanisms underlying leaf variegation of ‘Marble Queen’ differ from the closely related ‘Golden Pothos’. The established regeneration system and identified variants could be important materials for further investigation of leaf variegation in E. aureum.
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Thidiazuron promotes adventitious shoot regeneration from pothos (Epipremnum aureum) leaf and petiole explants
In Vitro Cellular & Developmental Biology - Plant, 2002Co-Authors: Jianjn Chen, Richard J Henny, Yingfeng Huang, Russell D. Caldwell, Cynthia A. RobinsonAbstract:Regeneration of adventitious shoots of pothos ( Epipremnum aureum Linden and Andre) ‘Jade’ was obtained using leaf and petiole explants preprated from shoot tips of 3-yr-old greenhouse-grown plants. Explants were cultured on Murashige and Skoog (MS) basal medium supplemented with thidiazuron (TDZ), 6-(4-hydroxy-3-methy-trans-2-butenyl-amino)purine (zeatin) or N -isopentenylaminopurine (2iP) individually with α-naphthaleneacetic acid (NAA) in 18 combinations. Callus was initiated from cut surface and along the midrib or major vein of leaf sections. Shoot regeneration from leaf and petoole explants occurred in 30d on medium containing 1, 5 or 10μ M TDZ with 0.5 or 1.0μ M NAA except petioles on medium with 10 μ M TDZ and 1.0 μ M NAA where regeneration failed. More time (50d) was needed for shoot regeneration when explants were cultured on medium containing either 2iP or zeatin with NAA. Regeneration frequencies were up to 20% and 50% for leaf and petiole explants, respectively. Shoot numbers per responding explant attained 30 for leaf and petiole explants on medium containing TDZ but only one to four on medium containing either 2iP or zeatin. These results indicate that TDZ is a more effective cytokinin for in vitro regeneration of pothos than either zeatin or 2iP.. Shoots elongated readily and rooted well on MS basal medium, without plant growth regulators. Plantlets acclimatized rapidly and grew vigorously in the greenhouse after transfer to pots containing a commerecial potting medium.
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glutathione dependent formaldehyde dehydrogenase from golden pothos Epipremnum aureum and the production of formaldehyde detoxifying plants
Plant Biotechnology, 2011Co-Authors: Yuichi Tada, Yusuke KiduAbstract:Glutathione-dependent formaldehyde dehydrogenase (FALDH) is an enzyme involved in formaldehyde metabolism in eukaryotes. FALDH cDNA was cloned from golden pothos, which is reported to effectively purify gaseous formaldehyde from enclosed room atmosphere. FALDH cDNAs from Arabidopsis, rice and golden pothos were overexpressed in transgenic Arabidopsis, and the enzyme activity was compared to determine the one most suitable for the molecular breeding of formaldehyde-detoxifying plants. The transgenic lines exhibited modified levels of the FALDH transcript, i.e. 10-800% compared to the endogenous transcript, due to either an overexpression or a cosuppression phenotype. The enzyme activity in the crude leaf extract was not proportionate, but did correlate with the transcription levels with certain exceptions. The FALDHs from the three plant species indicated similar enzymatic activity on average. The capacity to detoxify exogenous formaldehyde in the transformants with the FALDHs was determined at the whole plant level. Plants overexpressing FALDH from the three plant species displayed up to a 40% increase in their efficiency to take up exogenous formaldehyde as compared with the wild-type plants. On the other hand, no difference in the survival rate was observed among the transformants and wild type plants on formaldehyde-containing agar medium. These results show the FALDH from golden pothos to be similarly or more effective for detoxifying formaldehyde in transgenic plants compared with Arabidopsis and rice, and that this cDNA is applicable to the molecular breeding of formaldehyde detoxifying plants.
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isolation and characterization of formaldehyde responsive genes from golden pothos Epipremnum aureum
Plant Biotechnology, 2010Co-Authors: Yuichi Tada, Tomoya Matsuzaki, Yuya TanakaAbstract:Plants absorb and metabolize formaldehyde, a C-1 compound that is one of the main indoor air pollutants. To elucidate the molecular mechanism of formaldehyde metabolism in plants, we isolated formaldehyde-responsive genes from golden pothos by means of GeneFishing PCR. We focused on the immediate-early response genes following formaldehyde treatment. Two full length cDNA sequences corresponding to a putative class II chitinase and a hypothetical novel protein, which we termed DEG2, were generated by rapid amplification of cDNA ends (RACE). The chitinase, which we designated EaCHI1, was up-regulated in the leaves and stems, whereas DEG2 was up-regulated in the leaves and roots of formaldehyde-treated golden pothos. Phylogenetic analysis showed that putative class II chitinases were split into two groups, monocot and dicot. EaCHI1 belonged to the former group, occupying the most basal level among the monocot chitinases analyzed. The identification of chitinase as a formaldehyde-responsive gene suggests a novel physiological role for this enzyme in plant carbon metabolism and environmental responses. The DEG2 sequence was not similar to any known protein sequence.
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genetic transformation of golden pothos Epipremnum aureum mediated by agrobacterium tumefaciens
Plant Cell Tissue and Organ Culture, 2008Co-Authors: Katsutoshi Kotsuka, Yuichi TadaAbstract:To establish a procedure for Agrobacterium tumefaciens-mediated transformation of golden pothos (Epipremnum aureum) plants, the effects of selection antibiotics and the preculture period of stem explants before A. tumefaciens infection were examined. Explants were co-cultivated with A. tumefaciens EHA105, harboring the plasmid pGWB2/cGUS, on a somatic embryo-inducing medium supplemented with acetosyringone. Resulting transgenic somatic embryos were screened on an antibiotic selection medium, and the transgenic pothos plants were regenerated on a germination medium. Hygromycin was the optimum selection antibiotic tested. The preculture period significantly affected the transformation efficiency, with explants precultured for one-day showing the best efficiency (5–30%). Both transformed hygromycin-resistant embryos and regenerated plants showed β-glucuronidase activity. Southern blot analysis confirmed transgene integration into the pothos genome. This reproducible transformation system for golden pothos may enable the molecular breeding of this very common indoor plant.