A cell line, SHK, was derived from the kidney of spotted halibut Verasper variegates. The cell line was subcultured more than 40 passages in minimum essential medium (MEM) supplemented with fetal bovine serum (FBS...A cell line, SHK, was derived from the kidney of spotted halibut Verasper variegates. The cell line was subcultured more than 40 passages in minimum essential medium (MEM) supplemented with fetal bovine serum (FBS) and 10 ng ml4 basic fibroblast growth factor (bFGF). Cell morphology from primary culture and subculture was observed continuously by microscopy. The SHK cell line consisted predominantly of fibroblast-like cells. The cell line was able to grow between 20℃ and 30℃ with the optimum growth at 24℃ and with a reduced growth between 12℃ and 20℃. The growth rate of the cells increased as the proportion of FBS increased from 10% to 20% at 28℃ with optimum growth at the concentration of 20%. The doubling time of the cells was determined to be 44.8 h. Chromosome analysis revealed that 52% of the SHK cells maintained a normal diploid chromosome number (2n=46). The cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies. The cells were infected by lymphosystis disease virus (LCDV) and found to be susceptible to the virus in cytopathic effect (CPE) observation. The infection was confirmed by PCR and electron microscopy experiments, which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.展开更多
Female grasshoppers can affect the fitness of their offspring through their selection of oviposition site. Knowledge of soil type on oviposition, and its effects on subsequent development can provide guidelines for ha...Female grasshoppers can affect the fitness of their offspring through their selection of oviposition site. Knowledge of soil type on oviposition, and its effects on subsequent development can provide guidelines for habitat manipulations that reduce the harmful effects of these pests on farmers fields. The influence of soil types on the oviposition site preference of variegated grasshopper (Zonocerus variegatus L.) reared some cassava (Manihot esculenta Crantz) varieties, was investigated in a cage trial carried out at the Bio factory laboratory, School of Agriculture and Food Sciences, Njala University, Sierra Leone during 2022/2023. The treatments comprised three soil types (Sandy, Loamy and Clay), each with three replications laid out in a randomized complete block design (RCBD) in wooden cages. Data were collected on the following development parameters including, Net reproductive growth ratio (R0), Generation time (Tc), Intrinsic rate of increase (rm), Finite rate of increase (), Doubling time (Dt), and overall survivorship. Findings revealed that, Z. variegatus L. preferred sandy soil in which, on average, most eggs were deposited (338, 6.62 4.40), followed by loamy soil, 286 (5.53 3.96), and then, clayey soil, 200 (3.91 3.85);though, the differences were not significant. This study established that Z. variegatus deposited more eggs in sandy soil > loamy soil > clayey soil, respectively;and subsequent survivorship of the immature unto mature adult insect, revealed a similar order. This indicates that the sandy soil is the most preferred substrate for oviposition and subsequent development into adult insects.展开更多
Isolated protoplasts from thalli of Porphyra haitanensis and Porphyra yezoensis were treated with colchicine or irradiated by ultraviolet (UV ). Several types of color variants were observed among the protoplast offsp...Isolated protoplasts from thalli of Porphyra haitanensis and Porphyra yezoensis were treated with colchicine or irradiated by ultraviolet (UV ). Several types of color variants were observed among the protoplast offspring. After treatment with colchicine: (1) 0.04-0.09% of red type variants in P. haitanensis were obtained; (2) The rate of red type variants and the variegated chimeral thalli composed of red type and wild type of sectors were 6.31- 1.11% in P. yezoensis. After irradiation with UV: (1) 3.5- 10.5% of red type variants in P. yezoensis were obtained: (2) 0.5-2-0% of red type variants and the variegated chimeral thalli composed of red type and wild type of sectors were obtained in P. haitanensis. Colchicine and UV’s mutangenic effects on P. yezoensis protoplasts were stronger than those on P. haitanensis protoplasts. The most efficient concentration of colchicine was 0.05%. The optimal length of UV-radiation was 1/2 min (radiation distance 5 cm). The red type variants induced, by colchicine展开更多
Leaf-color mutations have been studied extensively in plants.However,to better understand the complex mechanisms underlying the formation of leaf color,it is essential to continue discover novel genes involved in the ...Leaf-color mutations have been studied extensively in plants.However,to better understand the complex mechanisms underlying the formation of leaf color,it is essential to continue discover novel genes involved in the process of leaf color development.In this study,we identified a variegated-leaf(vg)mutant in tomato that exhibited defective phenotypes in thylakoids and photosynthesis.To clone the vg locus,an F2population was constructed from the cross between the vg mutant(Solanum lycopersicum)and the wild tomato LA1589(S.pimpinellifolium).Using the map-based cloning approach,the vg locus was mapped on chromosome 7 and narrowed down to a 128 kb region that contained 21 open reading frames(ORFs).The expression levels of ORF9,ORF10,and ORF13 were significantly lower in vg than in the wild-type plants,while the ORF11 transcript level was elevated in vg.We then mutagenized ORF9,ORF10,and ORF13 by the CRISPR/Cas9 system in the wild-type tomato background and found that only the ORF10 mutation reproduced the phenotype of variegated leaves,indicating that ORF10 represents VG and its down-regulated expression was responsible for the variegated leaf phenotype.ORF10 encodes a thylakoid formation protein and its mutant lines showed reduced levels of chlorophyll synthesis and photosynthesis.Taken together,these results suggest that VG is necessary for chloroplast development,chlorophyll synthesis,and photosynthesis in tomato.展开更多
Leaf variegation,the mosaic of colors on the leaf surface,can be developed by certain plant species without external influence.Although it may be associated with a variety of functions,the stable existence of differen...Leaf variegation,the mosaic of colors on the leaf surface,can be developed by certain plant species without external influence.Although it may be associated with a variety of functions,the stable existence of different leaf color morphs within a plant species has not been fully explained by previous studies.This study focuses on the two leaf morphs of Cypripedium forrestii,an endangered lady slipper orchid,and compares their micromorphological structure,photosynthetic potential,differentially expressed genes(DEGs),and ecological features to gain a comprehensive understanding of the underlying leaf variegation polymorphism.Our findings demonstrate that leaf variegation is not pathological and does not affect photosynthetic potential.Additionally,it significantly reduces herbivory damage.We found that the probability of herbivory and leaf area loss for variegated leaves was notably higher under drought conditions.Therefore,variegated individuals may be more adaptive under such conditions,while non-variegated ones may be more cost-effective in normal years.These results suggest that different leaf color morphs may be favored by varying environmental conditions,and leaf polymorphism may be a legacy of ancient climate and herbivore fluctuations.展开更多
Maize leaves are produced from polarized cell divisions that result in clonal cell lineages arrayed along the long axis of the leaf. We utilized this stereotypical division pattern to identify a collection of mutants ...Maize leaves are produced from polarized cell divisions that result in clonal cell lineages arrayed along the long axis of the leaf. We utilized this stereotypical division pattern to identify a collection of mutants that form chloroplast pigmentation sectors that violate the clonal cell lineages. Here, we describe the camouflage1 (cfl) mutant, which develops nonclonal, yellow-green sectors in its leaves. We cloned the cfl gene by transposon tagging and determined that it encodes porphobilinogen deaminase (PBGD), an enzyme that functions early in chlorophyll and heme biosynthesis. While PBGD has been characterized biochemically, no viable mutations in this gene have been reported in plants. To investigate the in vivo function of PBGD, we characterized the cfl mutant. Histological analyses revealed that cfl yellow sectors display the novel phenotype of bundle sheath cell-specific death. Light-shift experiments determined that constant light suppressed cfl sector formation, a dark/light transition is required to induce yellow sectors, and that sectors form only during a limited time of leaf development. Biochemical experiments determined that of 1 mutant leaves have decreased PBGD activity and increased levels of the enzyme substrate in both green and yellow regions. Furthermore, the cfl yellow regions displayed a reduction in catalase activity. A threshold model is hypothesized to explain the cfl variegation and incorporates photosynthetic cell differentiation, reactive oxygen species scavenging, and PBGD function.展开更多
Leaf variegation resulting from nuclear gene mutations has been used as a model system to elucidate the molecular mechanisms of chloroplast development. Since most variegation genes also function in photosynthesis, it...Leaf variegation resulting from nuclear gene mutations has been used as a model system to elucidate the molecular mechanisms of chloroplast development. Since most variegation genes also function in photosynthesis, it remains unknown whether their roles in photosynthesis and chloroplast development are distinct. Here, using the variegation mutant thylakoid formation1 (thfl) we show that variegation formation is light independent. It was found that slow and uneven chloroplast development in thfl can be attributed to defects in etioplast development in darkness. Ultrastructural analysis showed the coexistence of plastids with or without prolamellar bodies (PLB) in cells of thfl, but not of WT. Although THF1 mutation leads to significant decreases in the levels of Pchlide and Pchliide oxidoreductase (POR) expression, genetic and 5-aminolevulinic acid (ALA)-feeding analysis did not reveal Pchlide or POR to be critical factors for etioplast formation in thfl. Northern blot analysis showed that plastid gene expression is dramatically reduced in thfl compared with that in WT, particularly in the dark. Our results also indicate that chlorophyll biosynthesis and expression of plastidic genes are coordinately suppressed in thfl. Based on these results, we propose a model to explain leaf variegation formation from the plastid development perspective.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
VAR2 is an integral thylakoid membrane protein and a member of the versatile FtsH class of metalloproteases in prokaryotes and eukaryotes. Recessive mutations in the VAR2 locus give rise to variegated plants (var2) ...VAR2 is an integral thylakoid membrane protein and a member of the versatile FtsH class of metalloproteases in prokaryotes and eukaryotes. Recessive mutations in the VAR2 locus give rise to variegated plants (var2) that contain white sectors with abnormal plastids and green sectors with normal-appearing chloroplasts. In a continuing effort to isolate second-site suppressors of var2 variegation, we characterize in this report ems2505, a suppressor strain that has a virescent phenotype due to a missense mutation in At4g28590, the gene for a pioneer protein. We designated this gene SVR4 (for SUPPRESSOR OF VARIEGATION4) and the mutant allele in ems2505 as svr4-1. We demonstrate that SVR4 is located in chloroplasts and that svr4-1 single mutants are normal with respect to chloroplast anatomy and thylakoid membrane protein accumulation. However, they are modestly impaired in several aspects of photochemistry and have enhanced non-photochemical quenching (NPQ) capacity. A T-DNA insertion allele of SVR4, svr4-2, is seedling-lethal due to an early blockage of chloroplast development. We conclude that SVR4 is essential for chloroplast biogenesis, and hypothesize that SVR4 mediates some aspect of thylakoid structure or function that controls NPQ. We propose that in the suppressor strain, photoinhibitory pressure caused by a lack of VAR2 is ameliorated early in chloroplast development by enhanced NPQ capacity caused by reduced SVR4 activity. This would result in an increase in the number of chloroplasts that are able to surmount a threshold necessary to avoid photo-damage and thereby develop into functional chloroplasts.展开更多
The development of a plant leaf is a meticulously orchestrated sequence of events producing a complex organ comprising diverse cell types. The reticulate class of leaf variegation mutants displays contrasting pigmenta...The development of a plant leaf is a meticulously orchestrated sequence of events producing a complex organ comprising diverse cell types. The reticulate class of leaf variegation mutants displays contrasting pigmentation between veins and interveinal regions due to specific aberrations in the development of mesophyll cells. Thus, the reticulate mutants offer a potent tool to investigate cell-type-specific developmental processes. The discovery that most mutants are affected in plastid-localized, metabolic pathways that are strongly expressed in vasculature-associated tis- sues implicates a crucial role for the bundle sheath and their chloroplasts in proper development of the mesophyll cells. Here, we review the reticulate mutants and their phenotypic characteristics, with a focus on those in Arabidopsis thali- ana. Two alternative models have been put forward to explain the relationship between plastid metabolism and meso- phyll cell development, which we call here the supply and the signaling hypotheses. We critically assess these proposed models and discuss their implications for leaf development and bundle sheath function in C3 species. The characteriza- tion of the reticulate mutants supports the significance of plastid retrograde signaling in cell development and highlights the significance of the bundle sheath in C3 photosynthesis.展开更多
Arabidopsis thaliana L. yellow variegated (var2) mutant is defective in a chloroplast FtsH family metalloprotease, AtFtsH2/VAR2, and displays an intriguing green and white leaf variegation. This unique var2-mediated...Arabidopsis thaliana L. yellow variegated (var2) mutant is defective in a chloroplast FtsH family metalloprotease, AtFtsH2/VAR2, and displays an intriguing green and white leaf variegation. This unique var2-mediated leaf variegation offers a simple yet powerful tool for dissecting the genetic regulation of chloroplast development. Here, we report the isolation and characterization of a new var2 suppressor gene, SUPPRESSOR OF VARIEGA TION8 (SVR8), which encodes a putative chloroplast ribosomal large subunit protein, L24. Mutations in SVR8 suppress var2 leaf variegation at ambient temperature and partially suppress the cold-induced chlorosis phenotype of var2. Loss of SVR8 causes unique chloroplast rRNA processing defects, particularly the 23S-4.5S dicistronic precursor. The recovery of the major abnormal processing site in svr823S-4.5S precursor indicate that it does not lie in the same position where SVR8/L24 binds on the ribosome. Surprisingly, we found that the loss of a chloroplast ribosomal small subunit protein, $21, results in aberrant chloroplast rRNA processing but not suppression of var2 variegation. These findings suggest that the disruption of specific aspects of chloroplast translation, rather than a general impairment in chloroplast translation, suppress var2variegation and the existence of complex genetic interactions in chloroplast development.展开更多
On the morning of August 4,2024,the ChinaAustralia-New Zealand Buddhist Forum held three sub-forums at the University of Sydney,with themes respectively focusing on"Diverse Faiths,Variegated Cultures,and a Multic...On the morning of August 4,2024,the ChinaAustralia-New Zealand Buddhist Forum held three sub-forums at the University of Sydney,with themes respectively focusing on"Diverse Faiths,Variegated Cultures,and a Multicolour World:Constructing a Community of Shared Future for Mankind","The Interdependence Between Human and Nature",and"Chan Buddhism and the Modern World".展开更多
On August 2-3,2024,delegates from the Buddhist communities and academia of China,Australia and New Zealand,as well as other distinguished vips from all walks of life gathered in Sydney,Australia for the China-Austra...On August 2-3,2024,delegates from the Buddhist communities and academia of China,Australia and New Zealand,as well as other distinguished vips from all walks of life gathered in Sydney,Australia for the China-Australia-New Zealand Buddhist Forum.The forum centered on the theme of"Compassionate Harmony and Multicultural Coexistence"and conducted in-depth discussions on the main topics of"Diverse Faiths,Variegated Cultures and a Multicolour World:Constructing a Community of Shared Future for Mankind","Chan Buddhism and the Modern World"and"he Interdependence Between Human and Nature".The forum built a consensus and achieved positive results on these important issues.展开更多
基金supported by grants from State 863High-Technology Rand Project of China(2006AA09Z406,2006AA10A401)Taishan Scholar Project of Shan-dong Province
文摘A cell line, SHK, was derived from the kidney of spotted halibut Verasper variegates. The cell line was subcultured more than 40 passages in minimum essential medium (MEM) supplemented with fetal bovine serum (FBS) and 10 ng ml4 basic fibroblast growth factor (bFGF). Cell morphology from primary culture and subculture was observed continuously by microscopy. The SHK cell line consisted predominantly of fibroblast-like cells. The cell line was able to grow between 20℃ and 30℃ with the optimum growth at 24℃ and with a reduced growth between 12℃ and 20℃. The growth rate of the cells increased as the proportion of FBS increased from 10% to 20% at 28℃ with optimum growth at the concentration of 20%. The doubling time of the cells was determined to be 44.8 h. Chromosome analysis revealed that 52% of the SHK cells maintained a normal diploid chromosome number (2n=46). The cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies. The cells were infected by lymphosystis disease virus (LCDV) and found to be susceptible to the virus in cytopathic effect (CPE) observation. The infection was confirmed by PCR and electron microscopy experiments, which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.
文摘Female grasshoppers can affect the fitness of their offspring through their selection of oviposition site. Knowledge of soil type on oviposition, and its effects on subsequent development can provide guidelines for habitat manipulations that reduce the harmful effects of these pests on farmers fields. The influence of soil types on the oviposition site preference of variegated grasshopper (Zonocerus variegatus L.) reared some cassava (Manihot esculenta Crantz) varieties, was investigated in a cage trial carried out at the Bio factory laboratory, School of Agriculture and Food Sciences, Njala University, Sierra Leone during 2022/2023. The treatments comprised three soil types (Sandy, Loamy and Clay), each with three replications laid out in a randomized complete block design (RCBD) in wooden cages. Data were collected on the following development parameters including, Net reproductive growth ratio (R0), Generation time (Tc), Intrinsic rate of increase (rm), Finite rate of increase (), Doubling time (Dt), and overall survivorship. Findings revealed that, Z. variegatus L. preferred sandy soil in which, on average, most eggs were deposited (338, 6.62 4.40), followed by loamy soil, 286 (5.53 3.96), and then, clayey soil, 200 (3.91 3.85);though, the differences were not significant. This study established that Z. variegatus deposited more eggs in sandy soil > loamy soil > clayey soil, respectively;and subsequent survivorship of the immature unto mature adult insect, revealed a similar order. This indicates that the sandy soil is the most preferred substrate for oviposition and subsequent development into adult insects.
文摘Isolated protoplasts from thalli of Porphyra haitanensis and Porphyra yezoensis were treated with colchicine or irradiated by ultraviolet (UV ). Several types of color variants were observed among the protoplast offspring. After treatment with colchicine: (1) 0.04-0.09% of red type variants in P. haitanensis were obtained; (2) The rate of red type variants and the variegated chimeral thalli composed of red type and wild type of sectors were 6.31- 1.11% in P. yezoensis. After irradiation with UV: (1) 3.5- 10.5% of red type variants in P. yezoensis were obtained: (2) 0.5-2-0% of red type variants and the variegated chimeral thalli composed of red type and wild type of sectors were obtained in P. haitanensis. Colchicine and UV’s mutangenic effects on P. yezoensis protoplasts were stronger than those on P. haitanensis protoplasts. The most efficient concentration of colchicine was 0.05%. The optimal length of UV-radiation was 1/2 min (radiation distance 5 cm). The red type variants induced, by colchicine
基金supported by the National Natural Science Foundation of China(Grant Nos.31672149,31772317,and 32072595)China Postdoctoral Science Foundation(Grant No.2021M691174)。
文摘Leaf-color mutations have been studied extensively in plants.However,to better understand the complex mechanisms underlying the formation of leaf color,it is essential to continue discover novel genes involved in the process of leaf color development.In this study,we identified a variegated-leaf(vg)mutant in tomato that exhibited defective phenotypes in thylakoids and photosynthesis.To clone the vg locus,an F2population was constructed from the cross between the vg mutant(Solanum lycopersicum)and the wild tomato LA1589(S.pimpinellifolium).Using the map-based cloning approach,the vg locus was mapped on chromosome 7 and narrowed down to a 128 kb region that contained 21 open reading frames(ORFs).The expression levels of ORF9,ORF10,and ORF13 were significantly lower in vg than in the wild-type plants,while the ORF11 transcript level was elevated in vg.We then mutagenized ORF9,ORF10,and ORF13 by the CRISPR/Cas9 system in the wild-type tomato background and found that only the ORF10 mutation reproduced the phenotype of variegated leaves,indicating that ORF10 represents VG and its down-regulated expression was responsible for the variegated leaf phenotype.ORF10 encodes a thylakoid formation protein and its mutant lines showed reduced levels of chlorophyll synthesis and photosynthesis.Taken together,these results suggest that VG is necessary for chloroplast development,chlorophyll synthesis,and photosynthesis in tomato.
基金supported by:Science&Technology Basic Resources Investigation Program of China for Survey and Germplasm Conservation of PSESP in Southwest China(2017FY100100)the Second Tibetan Plateau Scientific Expedition and Research Program(2019QZKK0502)+1 种基金the PSESP project of Yunnan Forestry and Grassland Bureau(2021SJ14X-09)and the project“Collection and Conservation of Plant Species with Extremely Small Populations of Polystichum glaciale and Cypripedium forrestii in Lijiang”(2021SJ14X-11).
文摘Leaf variegation,the mosaic of colors on the leaf surface,can be developed by certain plant species without external influence.Although it may be associated with a variety of functions,the stable existence of different leaf color morphs within a plant species has not been fully explained by previous studies.This study focuses on the two leaf morphs of Cypripedium forrestii,an endangered lady slipper orchid,and compares their micromorphological structure,photosynthetic potential,differentially expressed genes(DEGs),and ecological features to gain a comprehensive understanding of the underlying leaf variegation polymorphism.Our findings demonstrate that leaf variegation is not pathological and does not affect photosynthetic potential.Additionally,it significantly reduces herbivory damage.We found that the probability of herbivory and leaf area loss for variegated leaves was notably higher under drought conditions.Therefore,variegated individuals may be more adaptive under such conditions,while non-variegated ones may be more cost-effective in normal years.These results suggest that different leaf color morphs may be favored by varying environmental conditions,and leaf polymorphism may be a legacy of ancient climate and herbivore fluctuations.
文摘Maize leaves are produced from polarized cell divisions that result in clonal cell lineages arrayed along the long axis of the leaf. We utilized this stereotypical division pattern to identify a collection of mutants that form chloroplast pigmentation sectors that violate the clonal cell lineages. Here, we describe the camouflage1 (cfl) mutant, which develops nonclonal, yellow-green sectors in its leaves. We cloned the cfl gene by transposon tagging and determined that it encodes porphobilinogen deaminase (PBGD), an enzyme that functions early in chlorophyll and heme biosynthesis. While PBGD has been characterized biochemically, no viable mutations in this gene have been reported in plants. To investigate the in vivo function of PBGD, we characterized the cfl mutant. Histological analyses revealed that cfl yellow sectors display the novel phenotype of bundle sheath cell-specific death. Light-shift experiments determined that constant light suppressed cfl sector formation, a dark/light transition is required to induce yellow sectors, and that sectors form only during a limited time of leaf development. Biochemical experiments determined that of 1 mutant leaves have decreased PBGD activity and increased levels of the enzyme substrate in both green and yellow regions. Furthermore, the cfl yellow regions displayed a reduction in catalase activity. A threshold model is hypothesized to explain the cfl variegation and incorporates photosynthetic cell differentiation, reactive oxygen species scavenging, and PBGD function.
基金supported by the Ministry of Science and Technology of China (2007CB108800 and 2009CB118504 to J. H.)Science and Technology Commission of Shanghai Municipality (09ZR1436300 to L. Z.)National Special Grantfor Transgenic Crops (2009ZX08009-081B to J. H.)
文摘Leaf variegation resulting from nuclear gene mutations has been used as a model system to elucidate the molecular mechanisms of chloroplast development. Since most variegation genes also function in photosynthesis, it remains unknown whether their roles in photosynthesis and chloroplast development are distinct. Here, using the variegation mutant thylakoid formation1 (thfl) we show that variegation formation is light independent. It was found that slow and uneven chloroplast development in thfl can be attributed to defects in etioplast development in darkness. Ultrastructural analysis showed the coexistence of plastids with or without prolamellar bodies (PLB) in cells of thfl, but not of WT. Although THF1 mutation leads to significant decreases in the levels of Pchlide and Pchliide oxidoreductase (POR) expression, genetic and 5-aminolevulinic acid (ALA)-feeding analysis did not reveal Pchlide or POR to be critical factors for etioplast formation in thfl. Northern blot analysis showed that plastid gene expression is dramatically reduced in thfl compared with that in WT, particularly in the dark. Our results also indicate that chlorophyll biosynthesis and expression of plastidic genes are coordinately suppressed in thfl. Based on these results, we propose a model to explain leaf variegation formation from the plastid development perspective.
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.
文摘VAR2 is an integral thylakoid membrane protein and a member of the versatile FtsH class of metalloproteases in prokaryotes and eukaryotes. Recessive mutations in the VAR2 locus give rise to variegated plants (var2) that contain white sectors with abnormal plastids and green sectors with normal-appearing chloroplasts. In a continuing effort to isolate second-site suppressors of var2 variegation, we characterize in this report ems2505, a suppressor strain that has a virescent phenotype due to a missense mutation in At4g28590, the gene for a pioneer protein. We designated this gene SVR4 (for SUPPRESSOR OF VARIEGATION4) and the mutant allele in ems2505 as svr4-1. We demonstrate that SVR4 is located in chloroplasts and that svr4-1 single mutants are normal with respect to chloroplast anatomy and thylakoid membrane protein accumulation. However, they are modestly impaired in several aspects of photochemistry and have enhanced non-photochemical quenching (NPQ) capacity. A T-DNA insertion allele of SVR4, svr4-2, is seedling-lethal due to an early blockage of chloroplast development. We conclude that SVR4 is essential for chloroplast biogenesis, and hypothesize that SVR4 mediates some aspect of thylakoid structure or function that controls NPQ. We propose that in the suppressor strain, photoinhibitory pressure caused by a lack of VAR2 is ameliorated early in chloroplast development by enhanced NPQ capacity caused by reduced SVR4 activity. This would result in an increase in the number of chloroplasts that are able to surmount a threshold necessary to avoid photo-damage and thereby develop into functional chloroplasts.
文摘The development of a plant leaf is a meticulously orchestrated sequence of events producing a complex organ comprising diverse cell types. The reticulate class of leaf variegation mutants displays contrasting pigmentation between veins and interveinal regions due to specific aberrations in the development of mesophyll cells. Thus, the reticulate mutants offer a potent tool to investigate cell-type-specific developmental processes. The discovery that most mutants are affected in plastid-localized, metabolic pathways that are strongly expressed in vasculature-associated tis- sues implicates a crucial role for the bundle sheath and their chloroplasts in proper development of the mesophyll cells. Here, we review the reticulate mutants and their phenotypic characteristics, with a focus on those in Arabidopsis thali- ana. Two alternative models have been put forward to explain the relationship between plastid metabolism and meso- phyll cell development, which we call here the supply and the signaling hypotheses. We critically assess these proposed models and discuss their implications for leaf development and bundle sheath function in C3 species. The characteriza- tion of the reticulate mutants supports the significance of plastid retrograde signaling in cell development and highlights the significance of the bundle sheath in C3 photosynthesis.
基金supported by funds from National Natural Science Foundation of China to F.Y. (31071073 31170219) and to X.L. (31100864)+2 种基金by Chinese Ministry of Education Program of New Century Excellent Talents in University (NCET 09 0657) to F.Yby funding to S.R. from the US Department of Energy Energy Biosciences panel (DE FG02 94ER20147)
文摘Arabidopsis thaliana L. yellow variegated (var2) mutant is defective in a chloroplast FtsH family metalloprotease, AtFtsH2/VAR2, and displays an intriguing green and white leaf variegation. This unique var2-mediated leaf variegation offers a simple yet powerful tool for dissecting the genetic regulation of chloroplast development. Here, we report the isolation and characterization of a new var2 suppressor gene, SUPPRESSOR OF VARIEGA TION8 (SVR8), which encodes a putative chloroplast ribosomal large subunit protein, L24. Mutations in SVR8 suppress var2 leaf variegation at ambient temperature and partially suppress the cold-induced chlorosis phenotype of var2. Loss of SVR8 causes unique chloroplast rRNA processing defects, particularly the 23S-4.5S dicistronic precursor. The recovery of the major abnormal processing site in svr823S-4.5S precursor indicate that it does not lie in the same position where SVR8/L24 binds on the ribosome. Surprisingly, we found that the loss of a chloroplast ribosomal small subunit protein, $21, results in aberrant chloroplast rRNA processing but not suppression of var2 variegation. These findings suggest that the disruption of specific aspects of chloroplast translation, rather than a general impairment in chloroplast translation, suppress var2variegation and the existence of complex genetic interactions in chloroplast development.
文摘On the morning of August 4,2024,the ChinaAustralia-New Zealand Buddhist Forum held three sub-forums at the University of Sydney,with themes respectively focusing on"Diverse Faiths,Variegated Cultures,and a Multicolour World:Constructing a Community of Shared Future for Mankind","The Interdependence Between Human and Nature",and"Chan Buddhism and the Modern World".
文摘On August 2-3,2024,delegates from the Buddhist communities and academia of China,Australia and New Zealand,as well as other distinguished vips from all walks of life gathered in Sydney,Australia for the China-Australia-New Zealand Buddhist Forum.The forum centered on the theme of"Compassionate Harmony and Multicultural Coexistence"and conducted in-depth discussions on the main topics of"Diverse Faiths,Variegated Cultures and a Multicolour World:Constructing a Community of Shared Future for Mankind","Chan Buddhism and the Modern World"and"he Interdependence Between Human and Nature".The forum built a consensus and achieved positive results on these important issues.