Recent advances in the proteomic field have allowed high-throughput experiments to be conducted on chloroplast samples. Many proteomic investigations have focused on either whole chloroplast or sub-plastidial fraction...Recent advances in the proteomic field have allowed high-throughput experiments to be conducted on chloroplast samples. Many proteomic investigations have focused on either whole chloroplast or sub-plastidial fractions. To date, the Plant Protein Database （PPDB, Sun et al., 2009） presents the most exhaustive chloroplast proteome available online. However, the accurate localization of many proteins that were identified in different sub-plastidial compartments remains hypothetical. Ferro et al. （2009） went a step further into the knowledge of Arabidopsis thaliana chloroplast proteins with regards to their accurate localization within the chloroplast by using a semi-quantitative proteomic approach known as spectral counting. Their proteomic strategy was based on the accurate mass and time tags （AMT） database approach and they built up AT_CHLORO, a comprehensive chloroplast proteome database with sub-plastidial localization and curated information on envelope proteins. Comparing these two extensive databases, we focus here on about 100 enzymes involved in the synthesis of chloroplast-specific isoprenoids. Well known pathways （i.e. compartmentation of the methyl erythritol phosphate biosynthetic pathway, of tetrapyrroles and chlorophyll biosynthesis and breakdown within chloroplasts） validate the spectral counting-based strategy. The same strategy was then used to identify the precise localization of the biosynthesis of carotenoids and prenylquinones within chloroplasts （i.e. in envelope membranes, stroma, and/or thylakoids） that remains unclear until now.展开更多
Chlorophyll(Chl) biosynthesis is essential for photosynthesis and plant growth.Glutamyl-tRNA reductase(GluTR) catalyzes glutamyl-tRNA into glutamate-1-semialdehyde(GSA) and initiates the chlorophyll biosynthesis.Even ...Chlorophyll(Chl) biosynthesis is essential for photosynthesis and plant growth.Glutamyl-tRNA reductase(GluTR) catalyzes glutamyl-tRNA into glutamate-1-semialdehyde(GSA) and initiates the chlorophyll biosynthesis.Even though the main role of GluTR has been established,the effects caused by natural variations in its corresponding gene remain largely unknown.Here,we characterized a spontaneous mutant in paddy field with Chl biosynthesis deficiency,designated as cbd1.With intact thylakoid lamellar structure,the cbd1 plant showed light green leaves and reduced Chl and carotenoids(Cars) content significantly compared to the wild type.By map-based gene cloning,the mutation was restricted within a 57-kb region on chromosome 10,in which an mPingA miniature inverted-repeat transposable element(MITE) inserted in the promoter region of OsHemA gene.Both leaf color and the pigment contents in cbd1 were recovered in a complementation test,confirming OsHemA was responsible for the mutant phenotype.OsHemA was uniquely predicted to encode GluTR and its expression level was dramatically repressed in cbd1.Transient transformation in protoplasts demonstrated that GluTR localized in chloroplasts and a signal peptide exists in its N-terminus.A majority of Chl biosynthesis genes,except for POR and CHLG,were down-regulated synchronously by the repression of OsHemA,suggesting that an attenuation occurred in the Chl biosynthesis pathway.Interestingly,we found major agronomic traits involved in rice yield were statistically unaffected,except for the number of full grains per panicle was increased in cbd1.Collectively,OsHemA plays an essential role in Chl biosynthesis in rice and its weak allele can adjust leaf color and Chls content without compromise to rice yield.展开更多
Chlorophyll biosynthesis is critical for chloroplast development and photosynthesis in plants. Although reactions in the chlorophyll biosynthetic pathway have been largely known, little is known about the regu-latory ...Chlorophyll biosynthesis is critical for chloroplast development and photosynthesis in plants. Although reactions in the chlorophyll biosynthetic pathway have been largely known, little is known about the regu-latory mechanisms of this pathway. In this study, we found that the dark-grown knockout and knockdown mutants as well as RNA-interference transgenic seedlings of BRAHMA （BRM）, which encodes an SWI2/ SNF2 chromatin-remodeling ATPase, had iligher greening rates, accumulated less protochlorophyllide, and produced less reactive oxygen species than Arabidopsis wild-type plants did upon light exposure. The expression of NADPH：protochlorophyilide oxidoreductase A （PORA）, PORB, and PORC, which catalyze a key step in chlorophyll biosynthesis, was increased in the brm mutants. We found that BRM physically interacted with the bHLH transcription factor PHYTOCHRONIE-iNTERACTING FACTOR 1 （PIF1） through its N-terminal domains. Furthermore, we demonstrated that BRM was directly recruited to the cis-regula-tory regions of PORC, but not of PORA and PORB, at least partially in a PIF1-dependent manner and the level of histone H3 lysine 4 tri-methylation （H3K4me3） at PORC loci was increased in the brm mutant. Taken together, our data indicate that the chromatino-remodeling enzyme BRM modulates PORC expression through interacting with PIF1, providing a novel regulatory mechanism by which plants fine-tune chloro-phyll biosynthesis during the transition from heterotrophic to autotrophic growth.展开更多
The effects of three types of antibiotics （erythromycin,ciprofloxacin and sulfamethoxazole） on the photosynthesis of freshwater algae,Selenastrum capricornutum Printz,were investigated by determining the growth rate...The effects of three types of antibiotics （erythromycin,ciprofloxacin and sulfamethoxazole） on the photosynthesis of freshwater algae,Selenastrum capricornutum Printz,were investigated by determining the growth rate,chloroplast pigments content,seven main precursors （including δ-aminolevulinic acid,porphobilinogen,uroporphyrinogen III,coproporphyrinogen III,protoporphyrin IX,Mg-proporphyrin IX and protochlorophyllide）,and photosynthetic rate during chlorophyll biosynthesis.The antibiotics significantly decreased the growth rate,chlorophyll content,and photosynthetic rate.Erythromycin induced a decreasing effect at a concentration of 0.06 mg/L,while ciprofloxacin and sulfamethoxazole achieved the same results at concentrations higher than 1.5 mg/L.Only erythromycin significantly inhibited chlorophyll biosynthesis,which indicated that it was considerably more toxic to S.capricornutum than ciprofloxacin and sulfamethoxazole,and may pose a high potential risk to aquatic ecosystems.展开更多
Light and chloroplast function is known to affect the plant immune response; however, the underlying mechanism remains elusive. We previously demonstrated that two light signaling factors, FAR-RED ELONGATED HYPOCOTYL ...Light and chloroplast function is known to affect the plant immune response; however, the underlying mechanism remains elusive. We previously demonstrated that two light signaling factors, FAR-RED ELONGATED HYPOCOTYL 3（FHY3）and FAR-RED IMPAIRED RESPONSE 1（FAR1）, regulate chlorophyll biosynthesis and seedling growth via controlling HEMB1 expression in Arabidopsis thaliana. In this study, we reveal that FHY3 and FAR1 are involved in modulating plant immunity. We showed that the fhy3 far1 double null mutant displayed high levels of reactive oxygen species and salicylic acid（SA） and increased resistance to Pseudomonas syringae pathogen infection. Microarray analysis revealed that a large proportion of pathogen-related genes, particularly genes encoding nucleotide-binding and leucine-rich repeat domain resistant proteins, are highly induced in fhy3 far1. Genetic studies indicated that the defects of fhy3 far1 can be largely rescued by reducing SA signaling or blocking SA accumulation, and by overexpression of HEMB1, which encodes a 5-aminolevulinic acid dehydratase in the chlorophyll biosynthetic pathway.Furthermore, we found that transgenic plants with reduced expression of HEMB1 exhibit a phenotype similar to fhy3 far1.Taken together, this study demonstrates an important role of FHY3 and FAR1 in regulating plant immunity, through integrating chlorophyll biosynthesis and the SA signaling pathway.展开更多
Green petals pose a challenge for pollinators to distinguish flowers from leaves,but they are valuable as a specialty flower trait.However,little is understood about the molecular mechanisms that underlie the developm...Green petals pose a challenge for pollinators to distinguish flowers from leaves,but they are valuable as a specialty flower trait.However,little is understood about the molecular mechanisms that underlie the development of green petals.Here,we report that CINCINNATA(CIN)-like TEOSINTE BRANCHED 1/CYCLOIDEA/PCF(TCP)proteins play key roles in the control of petal color.The septuple tcp2/3/4/5/10/13/17 mutant produced flowers with green petals due to chlorophyll accumulation.Expression of TCP4 complemented the petal phenotype of tcp2/3/4/5/10/13/17.We found that chloroplasts were converted into leucoplasts in the distal parts of wild-type petals but not in the proximal parts during flower development,whereas plastid conversion was compromised in the distal parts of tcp2/3/4/5/10/13/17 petals.TCP4 and most CIN-like TCPs were predominantly expressed in distal petal regions,consistent with the green–white pattern in wild-type petals and the petal greening observed in the distal parts of tcp2/3/4/5/10/13/17 petals.RNA-sequencing data revealed that most chlorophyll biosynthesis genes were downregulated in the white distal parts of wild-type petals,but these genes had elevated expression in the distal green parts of tcp2/3/4/5/10/13/17 petals and the green proximal parts of wild-type petals.We revealed that TCP4 repressed chlorophyll biosynthesis by directly binding to the promoters of PROTOCHLOROPHYLLIDE REDUCTASE(PORB),DIVINYL REDUCTASE(DVR),and SUPPRESSOR OF OVEREXPRESSION OF CO 1(SOC1),which are known to promote petal greening.We found that the conversion of chloroplasts to leucoplasts and the green coloration in the proximal parts of petals appeared to be conserved among plant species.Our findings uncover a major molecular mechanism that underpins the formation of petal color patterns and provide a foundation for the breeding of plants with green flowers.展开更多
文摘Recent advances in the proteomic field have allowed high-throughput experiments to be conducted on chloroplast samples. Many proteomic investigations have focused on either whole chloroplast or sub-plastidial fractions. To date, the Plant Protein Database （PPDB, Sun et al., 2009） presents the most exhaustive chloroplast proteome available online. However, the accurate localization of many proteins that were identified in different sub-plastidial compartments remains hypothetical. Ferro et al. （2009） went a step further into the knowledge of Arabidopsis thaliana chloroplast proteins with regards to their accurate localization within the chloroplast by using a semi-quantitative proteomic approach known as spectral counting. Their proteomic strategy was based on the accurate mass and time tags （AMT） database approach and they built up AT_CHLORO, a comprehensive chloroplast proteome database with sub-plastidial localization and curated information on envelope proteins. Comparing these two extensive databases, we focus here on about 100 enzymes involved in the synthesis of chloroplast-specific isoprenoids. Well known pathways （i.e. compartmentation of the methyl erythritol phosphate biosynthetic pathway, of tetrapyrroles and chlorophyll biosynthesis and breakdown within chloroplasts） validate the spectral counting-based strategy. The same strategy was then used to identify the precise localization of the biosynthesis of carotenoids and prenylquinones within chloroplasts （i.e. in envelope membranes, stroma, and/or thylakoids） that remains unclear until now.
基金supported by the National Key Research and Development Program of China(2016YFD0101801)the National Excellent Doctoral Dissertation of China(201262)+2 种基金the Key Laboratory of Biology,Genetics and Breeding of Japonica Rice in Mid-lower Yangtze River,Ministry of Agriculture and Rural Affairs,China,the Collaborative Innovation Center for Hybrid Rice in Yangtze River,China,and the Jiangsu Collaborative Innovation Center for Modern Crop Production,China,the National High-Tech R&D Program of China(2014AA10A603-15)the National Key Technologies R&D Program of China during the 12th Five-Year Plan period(2013BAD01B02-16)the Jiangsu Science and Technology Development Program,China(BE2014394 and BE2015363)
文摘Chlorophyll(Chl) biosynthesis is essential for photosynthesis and plant growth.Glutamyl-tRNA reductase(GluTR) catalyzes glutamyl-tRNA into glutamate-1-semialdehyde(GSA) and initiates the chlorophyll biosynthesis.Even though the main role of GluTR has been established,the effects caused by natural variations in its corresponding gene remain largely unknown.Here,we characterized a spontaneous mutant in paddy field with Chl biosynthesis deficiency,designated as cbd1.With intact thylakoid lamellar structure,the cbd1 plant showed light green leaves and reduced Chl and carotenoids(Cars) content significantly compared to the wild type.By map-based gene cloning,the mutation was restricted within a 57-kb region on chromosome 10,in which an mPingA miniature inverted-repeat transposable element(MITE) inserted in the promoter region of OsHemA gene.Both leaf color and the pigment contents in cbd1 were recovered in a complementation test,confirming OsHemA was responsible for the mutant phenotype.OsHemA was uniquely predicted to encode GluTR and its expression level was dramatically repressed in cbd1.Transient transformation in protoplasts demonstrated that GluTR localized in chloroplasts and a signal peptide exists in its N-terminus.A majority of Chl biosynthesis genes,except for POR and CHLG,were down-regulated synchronously by the repression of OsHemA,suggesting that an attenuation occurred in the Chl biosynthesis pathway.Interestingly,we found major agronomic traits involved in rice yield were statistically unaffected,except for the number of full grains per panicle was increased in cbd1.Collectively,OsHemA plays an essential role in Chl biosynthesis in rice and its weak allele can adjust leaf color and Chls content without compromise to rice yield.
文摘Chlorophyll biosynthesis is critical for chloroplast development and photosynthesis in plants. Although reactions in the chlorophyll biosynthetic pathway have been largely known, little is known about the regu-latory mechanisms of this pathway. In this study, we found that the dark-grown knockout and knockdown mutants as well as RNA-interference transgenic seedlings of BRAHMA （BRM）, which encodes an SWI2/ SNF2 chromatin-remodeling ATPase, had iligher greening rates, accumulated less protochlorophyllide, and produced less reactive oxygen species than Arabidopsis wild-type plants did upon light exposure. The expression of NADPH：protochlorophyilide oxidoreductase A （PORA）, PORB, and PORC, which catalyze a key step in chlorophyll biosynthesis, was increased in the brm mutants. We found that BRM physically interacted with the bHLH transcription factor PHYTOCHRONIE-iNTERACTING FACTOR 1 （PIF1） through its N-terminal domains. Furthermore, we demonstrated that BRM was directly recruited to the cis-regula-tory regions of PORC, but not of PORA and PORB, at least partially in a PIF1-dependent manner and the level of histone H3 lysine 4 tri-methylation （H3K4me3） at PORC loci was increased in the brm mutant. Taken together, our data indicate that the chromatino-remodeling enzyme BRM modulates PORC expression through interacting with PIF1, providing a novel regulatory mechanism by which plants fine-tune chloro-phyll biosynthesis during the transition from heterotrophic to autotrophic growth.
基金supported by the National Natural Science Foundation of China (No.40471118)the National Natural Science Foundation-Joint Founds of Guangdong Province (No.U0633006,U0633002)
文摘The effects of three types of antibiotics （erythromycin,ciprofloxacin and sulfamethoxazole） on the photosynthesis of freshwater algae,Selenastrum capricornutum Printz,were investigated by determining the growth rate,chloroplast pigments content,seven main precursors （including δ-aminolevulinic acid,porphobilinogen,uroporphyrinogen III,coproporphyrinogen III,protoporphyrin IX,Mg-proporphyrin IX and protochlorophyllide）,and photosynthetic rate during chlorophyll biosynthesis.The antibiotics significantly decreased the growth rate,chlorophyll content,and photosynthetic rate.Erythromycin induced a decreasing effect at a concentration of 0.06 mg/L,while ciprofloxacin and sulfamethoxazole achieved the same results at concentrations higher than 1.5 mg/L.Only erythromycin significantly inhibited chlorophyll biosynthesis,which indicated that it was considerably more toxic to S.capricornutum than ciprofloxacin and sulfamethoxazole,and may pose a high potential risk to aquatic ecosystems.
基金supported by grants from the National Natural Science Foundation of China(31170221,31325002 and 31300206)the Ministry of Agriculture of China(2014ZX08009-003)
文摘Light and chloroplast function is known to affect the plant immune response; however, the underlying mechanism remains elusive. We previously demonstrated that two light signaling factors, FAR-RED ELONGATED HYPOCOTYL 3（FHY3）and FAR-RED IMPAIRED RESPONSE 1（FAR1）, regulate chlorophyll biosynthesis and seedling growth via controlling HEMB1 expression in Arabidopsis thaliana. In this study, we reveal that FHY3 and FAR1 are involved in modulating plant immunity. We showed that the fhy3 far1 double null mutant displayed high levels of reactive oxygen species and salicylic acid（SA） and increased resistance to Pseudomonas syringae pathogen infection. Microarray analysis revealed that a large proportion of pathogen-related genes, particularly genes encoding nucleotide-binding and leucine-rich repeat domain resistant proteins, are highly induced in fhy3 far1. Genetic studies indicated that the defects of fhy3 far1 can be largely rescued by reducing SA signaling or blocking SA accumulation, and by overexpression of HEMB1, which encodes a 5-aminolevulinic acid dehydratase in the chlorophyll biosynthetic pathway.Furthermore, we found that transgenic plants with reduced expression of HEMB1 exhibit a phenotype similar to fhy3 far1.Taken together, this study demonstrates an important role of FHY3 and FAR1 in regulating plant immunity, through integrating chlorophyll biosynthesis and the SA signaling pathway.
基金supported by the National Science Fund for Distinguished Young Scholars of China(grant 31725005)the Science Fund for the Creative Research Groups of the National Natural Science Foundation of China(grant 31621001)the National Key R&D Program of China(2018YFE0204700).
文摘Green petals pose a challenge for pollinators to distinguish flowers from leaves,but they are valuable as a specialty flower trait.However,little is understood about the molecular mechanisms that underlie the development of green petals.Here,we report that CINCINNATA(CIN)-like TEOSINTE BRANCHED 1/CYCLOIDEA/PCF(TCP)proteins play key roles in the control of petal color.The septuple tcp2/3/4/5/10/13/17 mutant produced flowers with green petals due to chlorophyll accumulation.Expression of TCP4 complemented the petal phenotype of tcp2/3/4/5/10/13/17.We found that chloroplasts were converted into leucoplasts in the distal parts of wild-type petals but not in the proximal parts during flower development,whereas plastid conversion was compromised in the distal parts of tcp2/3/4/5/10/13/17 petals.TCP4 and most CIN-like TCPs were predominantly expressed in distal petal regions,consistent with the green–white pattern in wild-type petals and the petal greening observed in the distal parts of tcp2/3/4/5/10/13/17 petals.RNA-sequencing data revealed that most chlorophyll biosynthesis genes were downregulated in the white distal parts of wild-type petals,but these genes had elevated expression in the distal green parts of tcp2/3/4/5/10/13/17 petals and the green proximal parts of wild-type petals.We revealed that TCP4 repressed chlorophyll biosynthesis by directly binding to the promoters of PROTOCHLOROPHYLLIDE REDUCTASE(PORB),DIVINYL REDUCTASE(DVR),and SUPPRESSOR OF OVEREXPRESSION OF CO 1(SOC1),which are known to promote petal greening.We found that the conversion of chloroplasts to leucoplasts and the green coloration in the proximal parts of petals appeared to be conserved among plant species.Our findings uncover a major molecular mechanism that underpins the formation of petal color patterns and provide a foundation for the breeding of plants with green flowers.
