Pentatricopeptide repeat （PPR） proteins, charac- terized by tandem arrays of a 35 amino acid motif, have been suggested to play central and broad roles in modulating the expression of organelle genes in plants. Howe...Pentatricopeptide repeat （PPR） proteins, charac- terized by tandem arrays of a 35 amino acid motif, have been suggested to play central and broad roles in modulating the expression of organelle genes in plants. However, the molecular mechanisms of most rice PPR genes remains unclear. In this paper, we isolated and characterized a temperature-conditional virescent mutant, OsV4, in rice （Oryza sativa cultivar Jiahual （WT, japonica rice variety））. The mutant displays albino phenotype and abnormal chloroplasts at the three leaf stage, which gradually turns green after the four leaf stage at a low temperature （20℃）. But the mutant always develops green leaves and well-developed chloroplasts at a high temperature （32℃）. Genetic and molecular analyses uncovered that OsV4 encodes a novel chloroplast-targeted PPR protein including four PPR motifs. Further investigations show that the mutant phenotype is associated with changes in chlorophyll content and chloroplast development. The OsV4 transcripts only accumulate to high levels in young leaves, indicating that its expression is tissue-specific. In addition, transcript levels of some ribosomal components and plastid- encoded polymerase-dependent genes are dramatically re- duced in the albino mutants grown at 20℃. These findings suggest that OsV4 plays an important role during early chloroplast development under cold stress in rice.展开更多
Although the multiple organellar RNA editing factors （MORFs） in the plastids of Arabidopsis thaliana have been extensively studied, molecular details underlying how MORFs affect plant development in other species, p...Although the multiple organellar RNA editing factors （MORFs） in the plastids of Arabidopsis thaliana have been extensively studied, molecular details underlying how MORFs affect plant development in other species, particularly in rice, remain largely unknown. Here we describe the characterization of wspl, a rice mutant with white-stripe leaves and panicles. Notably, wspl exhibited nearly white immature panicles at the heading stage. Transmission electron microscopy analysis and chlorophyll content measurement re- veale i a chloroplast developmental defect and reduced chlorophyll accumulation in wspl. Positional cloning of WSP1 found a point mutation in OsO4g51280, whose putative product shares high sequence similarity with MORF proteins. Complementation experiments demonstrated that WSP1 was responsible for the variegated phenotypes of wspl. WSP1 is localized to chloroplasts and the point mutation in wspl affected the editing of multiple organellar RNA sites. Owing to the defect in plastid RNA editing, chloroplast ribosome biogenesis and ndhA splicing were also impaired in wspl, which may affect normal chloroplast development in the leaves and panicles at the heading stage. Together, our results demonstrate the importance of rice WSP1 protein in chloroplast development and broaden our knowledge about MORF family members in rice.展开更多
Albino mutants are useful genetic resource for studying chlorophyll biosynthesis and chloroplast development and cloning genes involved in these processes in plants. Here we report a novel rice mutant low temperature ...Albino mutants are useful genetic resource for studying chlorophyll biosynthesis and chloroplast development and cloning genes involved in these processes in plants. Here we report a novel rice mutant low temperature albino I （ltal） that showed albino leaves before 4-leaf stage when grown under temperature lower than 20℃, but developed normal green leaves under temperature higher than 24℃ or similar morphological phenotypes in dark as did the wild-type （WT）. Our analysis showed that the contents of chlorophylls and chlo- rophyll precursors were remarkably decreased in the Ital mutant under low temperature compared to WT. Transmission electron microscope observation revealed that chloroplasts were defectively developed in the albino ltal leaves, which lacked of well-stacked granum and contained less stroma lamellae. These results suggested that the ltal mutation may delay the light-induced thylakoid assembly under low temperature. Genetic analysis indicated that the albino phenotype was controlled by a single recessive locus. Through map-based approach, we finally located the Ltal gene to a region of 40.3 kb on the short arm of chromosome 11. There are 8 predicted open reading frames （ORFs） in this region and two of them were deleted in ltal genome compared with the WT genome. The further characterization of the Ltal gene would provide a good approach to uncover the novel molecular mechanisms involved in chloroplast development under low temperature stress.展开更多
Genic male sterility(GMS)is critical for heterosis utilization and hybrid seed production.Although GMS mutants and genes have been studied extensively in plants,it has remained unclear whether chloroplast-associated p...Genic male sterility(GMS)is critical for heterosis utilization and hybrid seed production.Although GMS mutants and genes have been studied extensively in plants,it has remained unclear whether chloroplast-associated photosynthetic and metabolic activities are involved in the regulation of anther development.In this study,we characterized the function of ZmMs33/ZmGPAT6,which encodes a member of the glycerol-3-phosphate acyltransferase(GPAT)family that catalyzes the first step of the glycerolipid synthetic pathway.We found that normal structure and function of endothecium(En)chloroplasts maintained by ZmMs33-mediated lipid biosynthesis in tapetal cells are crucial for maize anther development.ZmMs33 is expressed mainly in the tapetum at early anther developmental stages and critical for cell proliferation and expansion at late stages.Chloroplasts in En cells of wild-type anthers function as starch storage sites before stage 10 but as photosynthetic factories since stage 10 to enable starch metabolism and carbohydrate supply.Loss of ZmMs33 function inhibits the biosynthesis of glycolipids and phospholipids,which are major components of En chloroplast membranes,and disrupts the development and function of En chloroplasts,resulting in the formation of abnormal En chloroplasts containing numerous starch granules.Further analyses reveal that starch synthesis during the day and starch degradation at night are greatly suppressed in the mutant anthers,leading to carbon starvation and low energy status,as evidenced by low trehalose-6-phosphate content and a reduced ATP/AMP ratio.The energy sensor and inducer of autophagy,SnRK1,was activated to induce early and excessive autophagy,premature PCD,and metabolic reprogramming in tapetal cells,finally arresting the elongation and development of mutant anthers.Taken together,our results not only show that ZmMs33 is required for normal structure and function of En chloroplasts but also reveal that starch metabolism and photosynthetic activities of En chloroplasts at different devel展开更多
Carotenoids, a class of natural pigments found in all photosynthetic organisms, are involved in a variety of physiological processes, including coloration, photoprotection, biosynthesis of abscisic acid （ABA） and ch...Carotenoids, a class of natural pigments found in all photosynthetic organisms, are involved in a variety of physiological processes, including coloration, photoprotection, biosynthesis of abscisic acid （ABA） and chloroplast biogenesis. Although carotenoid biosynthesis has been well studied biochemically, the genetic basis of the pathway is not well understood. Here, we report the characterization of two allelic Arabidopsis mutants, spontaneous cell death1-1 （spcl-1） and spc1-2. The weak allele spc1-1 mutant showed characteristics of bleached leaves, accumulation of superoxide and mosaic cell death. The strong mutant allele spc1-2 caused a complete arrest of plant growth and development shortly after germination, leading to a seedling-lethal phenotype. Genetic and molecular analyses indicated that SPC1 encodes a putative ζ-carotene desaturase （ZDS） in the carotenoid biosynthesis pathway. Analysis of carotenoids revealed that several major carotenoid compounds downstream of SPC 1/ZDS were substantially reduced in spc1-1, suggesting that SPC 1 is a functional ZDS. Consistent with the downregulated expression of CAO and PORB, the chlorophyll content was decreased in spc1-1 plants. In addition, expression of Lhcb1. 1, Lhcbl. 4 and RbcS was absent in spc1-2, suggesting the possible involvement of carotenoids in the plastid-to-nucleus retrograde signaling. The spc1-1 mutant also displays an ABA-deficient phenotype that can be partially rescued by the externally supplied phytohormone. These results suggest that SPC1/ZDS is essential for biosynthesis of carotenoids and plays a crucial role in plant growth and development.展开更多
Chloroplasts are important for plant growth and development.RNA editing in chloroplast converts cytidines(Cs)to uridine s(Us)at specific transcript positions and provides a correction mechanism to restore conserved co...Chloroplasts are important for plant growth and development.RNA editing in chloroplast converts cytidines(Cs)to uridine s(Us)at specific transcript positions and provides a correction mechanism to restore conserved codons or creates start or stop codons.However,the underlined molecular mechanism is not yet fully unders tood.In the present study,we identi fied a thermo-sensi tive mutantin leaf color 1(tst1)and found that TSL1 is allelic to DELAYED GREENING 1(DG1).The mis sense mutation of DG1 in tsl1 mutant confers a high temperature sensitivity and impaired chloroplast development at an elevated ambient temperature in Arabidopsis.Subsequent analysis showed that chloroplast RNA editing at seve ral sites including accD-2568,ndhD-2,and petL-5 is impaired in tsl1 mutant plants grown at an elevated temperature.DG1 interacts with MORF2 and other proteins such as DYW1 and DYW2 involved in chloroplast RNA editing.In vitro RNA electrophoretic mobility shift assay demonstrated that DG1 binds to RNA targets such as accD,ndhD,and petL.Thus,our results revealed that DG1 is important for maintaining chloroplast mRNA editing in Arabidopsis.展开更多
Plants contain various factors that transiently interact with subunits or intermediates of the thylakoid multiprotein complexes, promoting their stable association and integration. Hence, assembly factors are essentia...Plants contain various factors that transiently interact with subunits or intermediates of the thylakoid multiprotein complexes, promoting their stable association and integration. Hence, assembly factors are essential for chloroplast development and the transition from heterotrophic to phototrophic growth. Snowy cotyledon 2 （SCO2） is a DNAJ-like protein involved in thylakoid membrane biogenesis and interacts with the light-harvesting chlorophyll-binding protein LHCBI. In Arabidopsis thaliana, SCO2 function was previ- ously reported to be restricted to cotyledons. Here we show that disruption of SC02 in Lotus japonicus results not only in paler cotyledons but also in variegated true leaves. Furthermore, smaller and pale- green true leaves can also be observed in A. thaliana sco2 （atsco2） mutants under short-day conditions. In both species, SCO2 is required for proper accumulation of PSlI-LHCll complexes. In contrast to other variegated mutants, inhibition of chloroplastic translation strongly affects L. japonicus sco2 mutant devel- opment and fails to suppress their variegated phenotype. Moreover, inactivation of the suppressor of variegation AtClpR1 in the atsco2 background results in an additive double-mutant phenotype with variegated true leaves. Taken together, our results indicate that SCO2 plays a distinct role in PSll assembly or repair and constitutes a novel factor involved in leaf variegation.展开更多
基金supported by Natural Science Foundation of China (no. 30971552)Shanghai Municipal Education Commission of China (no. 14YZ076)+2 种基金Shanghai Municipal Science and Technology Commission of China (10DZ2271800, 12ZR1422000)Leading Academic Discipline Project of Shanghai Municipal Education Commission (no. J50401)the Food Safety and Nutrition Program of Shanghai Normal University (DXL123)
文摘Pentatricopeptide repeat （PPR） proteins, charac- terized by tandem arrays of a 35 amino acid motif, have been suggested to play central and broad roles in modulating the expression of organelle genes in plants. However, the molecular mechanisms of most rice PPR genes remains unclear. In this paper, we isolated and characterized a temperature-conditional virescent mutant, OsV4, in rice （Oryza sativa cultivar Jiahual （WT, japonica rice variety））. The mutant displays albino phenotype and abnormal chloroplasts at the three leaf stage, which gradually turns green after the four leaf stage at a low temperature （20℃）. But the mutant always develops green leaves and well-developed chloroplasts at a high temperature （32℃）. Genetic and molecular analyses uncovered that OsV4 encodes a novel chloroplast-targeted PPR protein including four PPR motifs. Further investigations show that the mutant phenotype is associated with changes in chlorophyll content and chloroplast development. The OsV4 transcripts only accumulate to high levels in young leaves, indicating that its expression is tissue-specific. In addition, transcript levels of some ribosomal components and plastid- encoded polymerase-dependent genes are dramatically re- duced in the albino mutants grown at 20℃. These findings suggest that OsV4 plays an important role during early chloroplast development under cold stress in rice.
文摘Although the multiple organellar RNA editing factors （MORFs） in the plastids of Arabidopsis thaliana have been extensively studied, molecular details underlying how MORFs affect plant development in other species, particularly in rice, remain largely unknown. Here we describe the characterization of wspl, a rice mutant with white-stripe leaves and panicles. Notably, wspl exhibited nearly white immature panicles at the heading stage. Transmission electron microscopy analysis and chlorophyll content measurement re- veale i a chloroplast developmental defect and reduced chlorophyll accumulation in wspl. Positional cloning of WSP1 found a point mutation in OsO4g51280, whose putative product shares high sequence similarity with MORF proteins. Complementation experiments demonstrated that WSP1 was responsible for the variegated phenotypes of wspl. WSP1 is localized to chloroplasts and the point mutation in wspl affected the editing of multiple organellar RNA sites. Owing to the defect in plastid RNA editing, chloroplast ribosome biogenesis and ndhA splicing were also impaired in wspl, which may affect normal chloroplast development in the leaves and panicles at the heading stage. Together, our results demonstrate the importance of rice WSP1 protein in chloroplast development and broaden our knowledge about MORF family members in rice.
基金supported by the grants from the National Basic Research Program of China(No.2009CB119000)the Ministry of Agriculture of China for Transgenic Research (Nos.2011ZX08009-003,2011ZX08001-005)the National Science Foundation of China(Nos.31000094,30970246, 31100188 and 31161130533)
文摘Albino mutants are useful genetic resource for studying chlorophyll biosynthesis and chloroplast development and cloning genes involved in these processes in plants. Here we report a novel rice mutant low temperature albino I （ltal） that showed albino leaves before 4-leaf stage when grown under temperature lower than 20℃, but developed normal green leaves under temperature higher than 24℃ or similar morphological phenotypes in dark as did the wild-type （WT）. Our analysis showed that the contents of chlorophylls and chlo- rophyll precursors were remarkably decreased in the Ital mutant under low temperature compared to WT. Transmission electron microscope observation revealed that chloroplasts were defectively developed in the albino ltal leaves, which lacked of well-stacked granum and contained less stroma lamellae. These results suggested that the ltal mutation may delay the light-induced thylakoid assembly under low temperature. Genetic analysis indicated that the albino phenotype was controlled by a single recessive locus. Through map-based approach, we finally located the Ltal gene to a region of 40.3 kb on the short arm of chromosome 11. There are 8 predicted open reading frames （ORFs） in this region and two of them were deleted in ltal genome compared with the WT genome. The further characterization of the Ltal gene would provide a good approach to uncover the novel molecular mechanisms involved in chloroplast development under low temperature stress.
基金the National Key Research and Development Program of China(2017YFD0102001,2018YFD0100806,2017YFD0101201)the National Transgenic Major Program of China(2018ZX0801006B,2018ZX0800922B)+3 种基金the National Natural Science Foundation of China(31971958,31771875,31871702)the Fundamental Research Funds for the Central Universities of China(06500136)the"Ten Thousand Plan"-National High Level Talents Special Support Plan(to X.W.)the Beijing Science&Technology Plan Program(Z191100004019005).
文摘Genic male sterility(GMS)is critical for heterosis utilization and hybrid seed production.Although GMS mutants and genes have been studied extensively in plants,it has remained unclear whether chloroplast-associated photosynthetic and metabolic activities are involved in the regulation of anther development.In this study,we characterized the function of ZmMs33/ZmGPAT6,which encodes a member of the glycerol-3-phosphate acyltransferase(GPAT)family that catalyzes the first step of the glycerolipid synthetic pathway.We found that normal structure and function of endothecium(En)chloroplasts maintained by ZmMs33-mediated lipid biosynthesis in tapetal cells are crucial for maize anther development.ZmMs33 is expressed mainly in the tapetum at early anther developmental stages and critical for cell proliferation and expansion at late stages.Chloroplasts in En cells of wild-type anthers function as starch storage sites before stage 10 but as photosynthetic factories since stage 10 to enable starch metabolism and carbohydrate supply.Loss of ZmMs33 function inhibits the biosynthesis of glycolipids and phospholipids,which are major components of En chloroplast membranes,and disrupts the development and function of En chloroplasts,resulting in the formation of abnormal En chloroplasts containing numerous starch granules.Further analyses reveal that starch synthesis during the day and starch degradation at night are greatly suppressed in the mutant anthers,leading to carbon starvation and low energy status,as evidenced by low trehalose-6-phosphate content and a reduced ATP/AMP ratio.The energy sensor and inducer of autophagy,SnRK1,was activated to induce early and excessive autophagy,premature PCD,and metabolic reprogramming in tapetal cells,finally arresting the elongation and development of mutant anthers.Taken together,our results not only show that ZmMs33 is required for normal structure and function of En chloroplasts but also reveal that starch metabolism and photosynthetic activities of En chloroplasts at different devel
基金grants from National Natural Science Foundation of China （Grant Nos. 30330360, 30125025 , 30221002） Chinese Academy of Sciences （Grant No. KSCX2- YW-N-015）
文摘Carotenoids, a class of natural pigments found in all photosynthetic organisms, are involved in a variety of physiological processes, including coloration, photoprotection, biosynthesis of abscisic acid （ABA） and chloroplast biogenesis. Although carotenoid biosynthesis has been well studied biochemically, the genetic basis of the pathway is not well understood. Here, we report the characterization of two allelic Arabidopsis mutants, spontaneous cell death1-1 （spcl-1） and spc1-2. The weak allele spc1-1 mutant showed characteristics of bleached leaves, accumulation of superoxide and mosaic cell death. The strong mutant allele spc1-2 caused a complete arrest of plant growth and development shortly after germination, leading to a seedling-lethal phenotype. Genetic and molecular analyses indicated that SPC1 encodes a putative ζ-carotene desaturase （ZDS） in the carotenoid biosynthesis pathway. Analysis of carotenoids revealed that several major carotenoid compounds downstream of SPC 1/ZDS were substantially reduced in spc1-1, suggesting that SPC 1 is a functional ZDS. Consistent with the downregulated expression of CAO and PORB, the chlorophyll content was decreased in spc1-1 plants. In addition, expression of Lhcb1. 1, Lhcbl. 4 and RbcS was absent in spc1-2, suggesting the possible involvement of carotenoids in the plastid-to-nucleus retrograde signaling. The spc1-1 mutant also displays an ABA-deficient phenotype that can be partially rescued by the externally supplied phytohormone. These results suggest that SPC1/ZDS is essential for biosynthesis of carotenoids and plays a crucial role in plant growth and development.
基金financially supported by grants from the National Natural Science Foundation of China(Nos.31625004 and 31872653)the 111 Project(B14027)。
文摘Chloroplasts are important for plant growth and development.RNA editing in chloroplast converts cytidines(Cs)to uridine s(Us)at specific transcript positions and provides a correction mechanism to restore conserved codons or creates start or stop codons.However,the underlined molecular mechanism is not yet fully unders tood.In the present study,we identi fied a thermo-sensi tive mutantin leaf color 1(tst1)and found that TSL1 is allelic to DELAYED GREENING 1(DG1).The mis sense mutation of DG1 in tsl1 mutant confers a high temperature sensitivity and impaired chloroplast development at an elevated ambient temperature in Arabidopsis.Subsequent analysis showed that chloroplast RNA editing at seve ral sites including accD-2568,ndhD-2,and petL-5 is impaired in tsl1 mutant plants grown at an elevated temperature.DG1 interacts with MORF2 and other proteins such as DYW1 and DYW2 involved in chloroplast RNA editing.In vitro RNA electrophoretic mobility shift assay demonstrated that DG1 binds to RNA targets such as accD,ndhD,and petL.Thus,our results revealed that DG1 is important for maintaining chloroplast mRNA editing in Arabidopsis.
文摘Plants contain various factors that transiently interact with subunits or intermediates of the thylakoid multiprotein complexes, promoting their stable association and integration. Hence, assembly factors are essential for chloroplast development and the transition from heterotrophic to phototrophic growth. Snowy cotyledon 2 （SCO2） is a DNAJ-like protein involved in thylakoid membrane biogenesis and interacts with the light-harvesting chlorophyll-binding protein LHCBI. In Arabidopsis thaliana, SCO2 function was previ- ously reported to be restricted to cotyledons. Here we show that disruption of SC02 in Lotus japonicus results not only in paler cotyledons but also in variegated true leaves. Furthermore, smaller and pale- green true leaves can also be observed in A. thaliana sco2 （atsco2） mutants under short-day conditions. In both species, SCO2 is required for proper accumulation of PSlI-LHCll complexes. In contrast to other variegated mutants, inhibition of chloroplastic translation strongly affects L. japonicus sco2 mutant devel- opment and fails to suppress their variegated phenotype. Moreover, inactivation of the suppressor of variegation AtClpR1 in the atsco2 background results in an additive double-mutant phenotype with variegated true leaves. Taken together, our results indicate that SCO2 plays a distinct role in PSll assembly or repair and constitutes a novel factor involved in leaf variegation.
