Cytoplasmic male sterility (CMS) is a maternally inherited trait that suppresses the production of viable pollen. CMS is a useful biological tool for confinement strategies to facilitate coexistence of genetically mod...Cytoplasmic male sterility (CMS) is a maternally inherited trait that suppresses the production of viable pollen. CMS is a useful biological tool for confinement strategies to facilitate coexistence of genetically modified (GM) and non-GM crops in case where it is required. The trait is reversible and can be restored to fertility in the presence of nuclear restorer genes (Rf genes) and by environmental impacts. The aim of this study was to investigate the influence of the level of irrigation on the stability of CMS maize hybrids under defined greenhouse conditions. Additionally the combination of irrigation and air temperature was studied. Three CMS maize hybrids were grown with different levels of irrigation and in different temperature regimes. Tassel characteristics, pollen production and fertility were assessed. The CMS stability was high in hot air temperatures and decreased in lower temperatures. The level of irrigation had no major effect on the level of sterility. The extent of these phenomena was depending on the genotype of CMS maize and should be known before using CMS for coexistence purposes.展开更多
Cytoplasmic male sterility(CMS) is a maternally inherited trait that results in the failure to produce functional pollen.It was identified in many plants,and it is widely used to exploit heterosis.
The Nsa cytoplasmic male sterility(CMS) system confers stable male sterility and offers great potential for production of hybrid seeds in oilseed rape. However, genes responsible for male sterility in Nsa CMS have not...The Nsa cytoplasmic male sterility(CMS) system confers stable male sterility and offers great potential for production of hybrid seeds in oilseed rape. However, genes responsible for male sterility in Nsa CMS have not been identified. By mitochondrial genome sequencing of Nsa CMS and its maintainer line,we identified in an Nsa CMS line several chimeric genes encoding hypothetical proteins harboring transmembrane domains. One novel chimeric gene orf346 showed high identity with cox1 at the 50 terminal region and was co-transcribed with nad3 and rps12 genes. Transgenic plants of orf346 fused with or without mitochondrial targeting peptide conferred complete male sterility in Arabidopsis. ORF346 was mitochondrion-localized. Expression of orf346 in Escherichia coli inhibited bacterial growth, with excessive accumulation of reactive oxygen species and decreased ATP content. These results reveal a link between the newly identified mitochondrial gene orf346 and the abortion of Nsa CMS. Inadequate energy supply and excessive accumulation of reactive oxygen species may account for pollen abortion in Nsa CMS plants.展开更多
Pepper hybrid seeds production using male sterility could lower cost by reducing time and labour, and increase thegenetic purity of the F1 seeds. To investigate the genetics of fertility restoration of the Peterson cy...Pepper hybrid seeds production using male sterility could lower cost by reducing time and labour, and increase thegenetic purity of the F1 seeds. To investigate the genetics of fertility restoration of the Peterson cytoplasmic sterility inpepper, a doubled haploid population of 115 pepper lines obtained from anther culture of the F1 hybrid between YoloWonder (sterility maintainer line) and Perennial (fertility restorer line) and the parental lines were test-crossed by 77013A(a strict cytoplasmic-genic male sterile line). The fertility of the test-crossed lines was assessed in greenhouse and openfield with the following three criteria: pollen index (PI, visual estimation of pollen amount per flower), pollen number (PN,pollen counting under microscope), and seed number (SN, the number of seeds per fruit in open pollination). Correlationsbetween the each couple of criteria within, as well as between the cultivation methods ranged from 0.55 to 0.84. Analysisof variance showed that the genotype (DH line) and environment were the significant sources of variation of the fertility.Narrow sense of heritance of fertility restoration ranged from 0.38 to 0.92, depending on the criteria and environment. Thedistribution of the progeny was continuous between the parental genotypes indicating the quantitative inheritance offertility restoration. Inferred from segregation according to Snape et al.(1984), the number of segregating genes wasestimated to be that three to four genetic factors were involved in pollen traits (PI and PN) and five to eight genetic factorsin seed production (SN). The heredity analysis of the CMS will be helpful for understanding of the genetic mechanism ofthe fertility restoration and the exploitation of the CMS in hybrid seed production.展开更多
Plant cytoplasmic male sterility(CMS)is maternally inherited and often manifested as aborted pollen development,but the molecular basis of abortion remains to be identified.To facilitate an investigation of CMS in cot...Plant cytoplasmic male sterility(CMS)is maternally inherited and often manifested as aborted pollen development,but the molecular basis of abortion remains to be identified.To facilitate an investigation of CMS in cotton,the complete sequence of cotton mitochondrial(mt)genome for CMS-D2 line ZBA was determined.The mt genome was assembled as a single circular molecule with 634,036 bp in length.A total of 194 ORFs,36 protein-coding genes,six r RNAs,and 24 t RNAs were identified.Several chimeric genes encoding hypothetical proteins with transmembrane domains were identified.Among them,a previously unknown chimeric gene,orf610a,which is composed of atp1 and a 485-bp downstream sequence of unknown nature,was identified.RT-PCR and q RT-PCR validation indicated that orf610a was expressed specifically in a sterile line.Ectopic expression of orf610a in yeast resulted in excessive accumulation of reactive oxygen species and reduction in ATP content,in addition to inhibition of cellular growth.Transgenic A.thaliana overexpressing orf610a fused with a mitochondrial targeting peptide displayed partial male sterility.Interaction between ORF610a and the nuclear-encoded protein RD22 indicated an association between ORF610a and pollen abortion.Positive feedback during transcriptional regulation between nuclear regulatory factors and the mt CMS gene may account for the male sterility of ZBA.展开更多
Moderate stimuli in mitochondria improve wideranging stress adaptability in animals, but whether mitochondria play similar roles in plants is largely unknown. Here, we report the enhanced stress adaptability of S-type...Moderate stimuli in mitochondria improve wideranging stress adaptability in animals, but whether mitochondria play similar roles in plants is largely unknown. Here, we report the enhanced stress adaptability of S-type cytoplasmic male sterility(CMS-S) maize and its association with mild expression of sterilizing gene ORF355. A CMS-S maize line exhibited superior growth potential and higher yield than those of the near-isogenic N-type line in saline fields. Moderate expression of ORF355 induced the accumulation of reactive oxygen species and activated the cellular antioxidative defense system. This adaptive response was mediated by elevation of the nicotinamide adenine dinucleotide concentration and associated metabolic homeostasis. Metabolome analysis revealed broad metabolic changes in CMS-S lines, even in the absence of salinity stress. Metabolic products associated with amino acid metabolism and galactose metabolism were substantially changed, which underpinned the alteration of the antioxidative defense system in CMS-S plants. The results reveal the ORF355-mediated superior stress adaptability in CMS-S maize and might provide an important route to developing salt-tolerant maize varieties.展开更多
Coordination between mitochondria and the nucleus is crucial for fertility determination in plants with cytoplasmic male sterility(CMS).Using yeast one-hybrid screening,we identified a transcription factor,ZmDREB1.7,t...Coordination between mitochondria and the nucleus is crucial for fertility determination in plants with cytoplasmic male sterility(CMS).Using yeast one-hybrid screening,we identified a transcription factor,ZmDREB1.7,that is highly expressed in sterile microspores at the large vacuole stage and activates the expression of mitochondria-encoded CMS gene orf355.Δpro,a weak allele of ZmDREB1.7 with the loss of a key unfolded protein response(UPR)motif in the promoter,partially restores male fertility of CMS-S maize.ZmDREB1.7 expression increases rapidly in response to antimycin A treatment,but this response is attenuated in theΔpro allele.Furthermore,we found that expression of orf355 in mitochondria activates mitochondrial retrograde signaling,which in turn induces ZmDREB1.7 expression.Taken together,these findings demonstrate that positive-feedback transcriptional regulation between a nuclear regulator and a mitochondrial CMS gene determines male sterility in maize,providing new insights into nucleus-mitochondria communication in plants.展开更多
Cytoplasmic male sterility(CMS)is a powerful tool for the exploitation of hybrid heterosis and the study of signaling and interactions between the nucleus and the cytoplasm.C-type CMS(CMS-C)in maize has long been used...Cytoplasmic male sterility(CMS)is a powerful tool for the exploitation of hybrid heterosis and the study of signaling and interactions between the nucleus and the cytoplasm.C-type CMS(CMS-C)in maize has long been used in hybrid seed production,but the underlying sterility factor and its mechanism of action remain unclear.In this study,we demonstrate that the mitochondrial gene atp6c confers male sterility in CMS-C maize.The ATP6C protein shows stronger interactions with ATP8 and ATP9 than ATP6 during the assembly of F1F0-ATP synthase(F-type ATP synthase,ATPase),thereby reducing the quantity and activity of assem-bled F_(1)F_(o)-ATP synthase.By contrast,the quantity and activity of the F1'component are increased in CMS-C lines.Reduced F1F0-ATP synthase activity causes accumulation of excess protons in the inner membrane space of the mitochondria,triggering a burst of reactive oxygen species(ROS),premature programmed cell death of the tapetai cells,and pollen abortion.Collectively,our study identifies a chimeric mitochondrial gene(ATP6C)that causes CMS in maize and documents the contribution of ATP6C to F1F0-ATP synthase assembly,thereby providing novel insights into the molecular mechanisms of male sterility in plants.展开更多
Normal microsporogenesis is determined by both nuclear and mitochondrial genes. In maize C-type cytoplasmic male sterility, it is unclear how the development of meiocytes and microspores is affected by the mitochondri...Normal microsporogenesis is determined by both nuclear and mitochondrial genes. In maize C-type cytoplasmic male sterility, it is unclear how the development of meiocytes and microspores is affected by the mitochondrial sterility gene and the nuclear restorer gene. In this study, we sequenced the transcriptomes of single meiocytes(tetrad stage) and early mononucleate microspores from sterile and restorer lines. The numbers of expressed genes varied in individual cells and fewer than half of the expressed genes were common to the same cell types. Four comparisons revealed 3379 differentially expressed genes(DEGs), with 277 putatively associated with mitochondria, 226 encoding transcription factors,and 467 possibly targeted by RF4. KEGG analysis indicated that the DEGs in the two lines at the tetrad stage were involved predominantly in carbon metabolism and in amino acid biosynthesis and metabolism, whereas the DEGs during the transition from the tetrad stage to the early mononucleate stage were associated mostly with regulation of protein metabolism, fatty acid metabolism, and anatomical structure morphogenesis. Thus, meiocyte and microspore development was affected by the surrounding cells and the restorer gene, and the restorer gene helped restore the redox homeostasis of microspores and the normal cellular reconstruction during the transition.展开更多
The integrated use of molecular marker-assisted selection (MAS) and anther culture has potential to significantly increase efficiency in plant breeding;however, reports on this kind of practical use are very limited. ...The integrated use of molecular marker-assisted selection (MAS) and anther culture has potential to significantly increase efficiency in plant breeding;however, reports on this kind of practical use are very limited. In the present study, we report the development of cytoplasmic male sterile (CMS) maintainers with aroma, disease resistance and red-brown hulls, as an example of integration of MAS and anther culture in rice breeding. A high-resolution melting (HRM)-based functional molecular marker was developed for the red-brown hull trait caused by a unique mutation (rbh1) in OsCAD2. Functional molecular markers for genes of rice blast resistance (Pi2), aroma (fgr) and red-brown hull (rbh1) were used for precise genotyping of individual plants in the BC1 and BC2F2 populations derived from a cross between CMS maintainers Huaxiang B (pi2–/rbh1–/fgr–) and Rong 3B (Pi2+/RBH1+/Fgr+). A total of 89 doubled haploid (DH) lines were generated from selected BC2F2 plants (Pi2+/rbh1–/fgr–) by anther culture. Seven DH lines were subsequently selected as the potential new CMS maintainers based on their overall performance and high resistance to blast. Our study demonstrated that integration of MAS and anther culture significantly accelerated the development of CMS maintainers with multiple stacked genes.展开更多
Cytoplasmic male sterility(CMS)-restorer system is a useful tool to exploit heterosis in soybean.The major restorer gene for the M-type CMS is known as Rf-m,located in the 162.4-kb region on chromosome 16.Sequence ana...Cytoplasmic male sterility(CMS)-restorer system is a useful tool to exploit heterosis in soybean.The major restorer gene for the M-type CMS is known as Rf-m,located in the 162.4-kb region on chromosome 16.Sequence analysis has revealed that the Rf-m locus in Glycine max consists of seven penta tricopeptide repeat(GmPPR)genes.The deduced amino acid sequences contain 8 to 14 PPR motifs,and a phylogenetic analysis grouped these GmPPR proteins into two PPR subfamilies:Glyma.16G161800 belongs to the PLS subfamily,and the P subfamily consists.of Glyma.16G161900,Glyma 16G162000,Glyma.16G162100,Glyma.16G162700,Glyma.16G162800,and Gly-ma 16G163100.The phylogenetic analysis of seven GmPPR proteins and 27 other plant PPR proteins also showed that proteins in the same subfamilies cluster together.Comparative sequence analysis was conducted using the seven Rf-m candidate GmPPR genes from the sterile line W931A,the maintainer line W931B,and the restorer line WR016,the result showed that Glyma 16G161900 had higher polymorphism than the other candidate genes.Based on real-time quantitative RT-PCR data,all seven GmPPR genes were differentially expressed but showed constitutive expression in roots,stems,leaves,and pollen grains.Additionally,the expression level of Gly-ma 16G161900 in the sterile line W931 A was significantly higher in all tissues than in the restorer line WR016.Taken together,these results suggest that Glyma 16G161900 is the most likely candidate for the restorer gene Rf-m.This study is the first report and analysis of candidate fertility restorer(Rf)genes encoding PPR proteins in soybean.展开更多
Isolation of high-quality mitochondrial DNA(mtDNA) is an important premise for researching molecular mechanisms in cytoplasmic male sterility of cabbage(Brassica oleracea L.var.capitata). An efficient protocol for sep...Isolation of high-quality mitochondrial DNA(mtDNA) is an important premise for researching molecular mechanisms in cytoplasmic male sterility of cabbage(Brassica oleracea L.var.capitata). An efficient protocol for separation and purification of mitochondria and extraction of mitochondrial DNA(mtDNA) from etiolated tissues of cabbage was developed. We took a method combined mannitol density gradient with differential centrifugation, selected appropriate rotational speed, extended DNase I treating time and changed mitochondria cracking condition. The results showed that the extracted mitochondria in this protocol had complete structure, appeared to ellipsoid and had not been contaminated with other impurities under the Jannus Green B staining. The isolated mitochondrial DNA had high purity and yield through detecting the optical density, nuclear specific primer PCR and agarose gel electrophoresis. The results indicated that mitochondrial DNA extracted by this protocol had high quality and enabled to be used in futher genetic studies.展开更多
The male sterility(MS)line is a prerequisite for efficient production of hybrid seeds in rice,a self-pollinating species.MS line breeding is pivotal for hybrid rice improvement.Understanding the historical breeding tr...The male sterility(MS)line is a prerequisite for efficient production of hybrid seeds in rice,a self-pollinating species.MS line breeding is pivotal for hybrid rice improvement.Understanding the historical breeding tra-jectory will help to improve hybrid rice breeding strategies.Maternally inherited cytoplasm is an appro-priate tool for phylogenetic reconstruction and pedigree tracing in rice hybrids.In this study,we analyzed the cytoplasmic genomes of 1495 elite hybrid rice varieties and identified five major types of cytoplasm,which correspond to different hybrid production systems.As the cytoplasm donors for hybrids,461 MS lines were also divided into five major types based on cytoplasmic and nuclear genomic architecture.Spe-cific core accessions cooperating with different fertility-associated genes drove the sequence divergence of MS lines.Dozens to hundreds of convergent and divergent selective sweeps spanning several agro-nomic trait-associated genes were identified among different types of MS lines.We further analyzed the cross patterns between different types of MS lines and their corresponding restorers.This study system-atically analyzed the cytoplasmic genomes of rice hybrids revealed their relationships with nuclear ge-nomes of MS lines,and illustrated the trajectory of hybrid rice breeding and the strategies for breeding different types of MS lines providing new insights for future improvement of hybrid rice.展开更多
Ogura cytoplasmic male sterility(Ogura CMS)is extensively applied in hybrid seed production in cruciferous crops.However,the posttranscriptional molecular basis of Ogura CMS in cruciferous crops remains elusive.Here,a...Ogura cytoplasmic male sterility(Ogura CMS)is extensively applied in hybrid seed production in cruciferous crops.However,the posttranscriptional molecular basis of Ogura CMS in cruciferous crops remains elusive.Here,a data-independent acquisition-based proteomic approach coupled with a parallel reaction monitoring-based targeted proteomic assay was used to analyze the proteome dynamics of Ogura CMS cabbage line RM and its maintainer line RF during floral bud development to obtain insights into the mechanism underlying Ogura CMS in cruciferous crops.A total of 9162 proteins corresponding to 61464 peptides were identified in RM and RF floral buds.The proteomic fluctuation of RM was weaker than that of RF.Differences in protein expression between RM and RF gradually enlarged with floral bud development.Fifteen continually up-regulated and eight continually down-regulated proteins were found in RM relative to RF throughout floral bud development.Differentially expressed proteins between RM and RF during floral bud development were implicated in the endoplasmic reticulum(ER)-associated protein processing pathway,in which most of them exhibited down-regulated expression in RM.These data suggest that ER-associated protein processing may be involved in pollen abortion in Ogura CMS cabbage by inhibiting the expression of critical factors.Our findings not only deepen the understanding of the molecular mechanisms of Ogura CMS in cruciferous crops but also provide better guidance for applying Ogura CMS in the hybrid breeding of cruciferous crops.展开更多
文摘Cytoplasmic male sterility (CMS) is a maternally inherited trait that suppresses the production of viable pollen. CMS is a useful biological tool for confinement strategies to facilitate coexistence of genetically modified (GM) and non-GM crops in case where it is required. The trait is reversible and can be restored to fertility in the presence of nuclear restorer genes (Rf genes) and by environmental impacts. The aim of this study was to investigate the influence of the level of irrigation on the stability of CMS maize hybrids under defined greenhouse conditions. Additionally the combination of irrigation and air temperature was studied. Three CMS maize hybrids were grown with different levels of irrigation and in different temperature regimes. Tassel characteristics, pollen production and fertility were assessed. The CMS stability was high in hot air temperatures and decreased in lower temperatures. The level of irrigation had no major effect on the level of sterility. The extent of these phenomena was depending on the genotype of CMS maize and should be known before using CMS for coexistence purposes.
文摘Cytoplasmic male sterility(CMS) is a maternally inherited trait that results in the failure to produce functional pollen.It was identified in many plants,and it is widely used to exploit heterosis.
基金supported by the National Key Research and Development Program of China (2016YFD0101300)the Natural Science Foundation of China (30871553)+3 种基金the Fundamental Research Funds for Central Nonprofit Scientific Institution (1610172017005)the Agricultural Science and Technology Innovation Program of CAAS (Group No. 118)the Hubei Agricultural Science and Technology Innovation Center (201620000001048)the China Agriculture Research System (CARS-12)。
文摘The Nsa cytoplasmic male sterility(CMS) system confers stable male sterility and offers great potential for production of hybrid seeds in oilseed rape. However, genes responsible for male sterility in Nsa CMS have not been identified. By mitochondrial genome sequencing of Nsa CMS and its maintainer line,we identified in an Nsa CMS line several chimeric genes encoding hypothetical proteins harboring transmembrane domains. One novel chimeric gene orf346 showed high identity with cox1 at the 50 terminal region and was co-transcribed with nad3 and rps12 genes. Transgenic plants of orf346 fused with or without mitochondrial targeting peptide conferred complete male sterility in Arabidopsis. ORF346 was mitochondrion-localized. Expression of orf346 in Escherichia coli inhibited bacterial growth, with excessive accumulation of reactive oxygen species and decreased ATP content. These results reveal a link between the newly identified mitochondrial gene orf346 and the abortion of Nsa CMS. Inadequate energy supply and excessive accumulation of reactive oxygen species may account for pollen abortion in Nsa CMS plants.
基金The study was funded by the National 863 Program, China （2002AA207012-1-3, 2001AA241121-9）the National Natural Science Foundation of China （3980453）.
文摘Pepper hybrid seeds production using male sterility could lower cost by reducing time and labour, and increase thegenetic purity of the F1 seeds. To investigate the genetics of fertility restoration of the Peterson cytoplasmic sterility inpepper, a doubled haploid population of 115 pepper lines obtained from anther culture of the F1 hybrid between YoloWonder (sterility maintainer line) and Perennial (fertility restorer line) and the parental lines were test-crossed by 77013A(a strict cytoplasmic-genic male sterile line). The fertility of the test-crossed lines was assessed in greenhouse and openfield with the following three criteria: pollen index (PI, visual estimation of pollen amount per flower), pollen number (PN,pollen counting under microscope), and seed number (SN, the number of seeds per fruit in open pollination). Correlationsbetween the each couple of criteria within, as well as between the cultivation methods ranged from 0.55 to 0.84. Analysisof variance showed that the genotype (DH line) and environment were the significant sources of variation of the fertility.Narrow sense of heritance of fertility restoration ranged from 0.38 to 0.92, depending on the criteria and environment. Thedistribution of the progeny was continuous between the parental genotypes indicating the quantitative inheritance offertility restoration. Inferred from segregation according to Snape et al.(1984), the number of segregating genes wasestimated to be that three to four genetic factors were involved in pollen traits (PI and PN) and five to eight genetic factorsin seed production (SN). The heredity analysis of the CMS will be helpful for understanding of the genetic mechanism ofthe fertility restoration and the exploitation of the CMS in hybrid seed production.
基金supported by funds from the National Natural Science Foundation of China(31871679)the Tianshan Youth Program(2018Q010)the Central Public-interest Scientific Institution Basal Research Fund(1610162021015)。
文摘Plant cytoplasmic male sterility(CMS)is maternally inherited and often manifested as aborted pollen development,but the molecular basis of abortion remains to be identified.To facilitate an investigation of CMS in cotton,the complete sequence of cotton mitochondrial(mt)genome for CMS-D2 line ZBA was determined.The mt genome was assembled as a single circular molecule with 634,036 bp in length.A total of 194 ORFs,36 protein-coding genes,six r RNAs,and 24 t RNAs were identified.Several chimeric genes encoding hypothetical proteins with transmembrane domains were identified.Among them,a previously unknown chimeric gene,orf610a,which is composed of atp1 and a 485-bp downstream sequence of unknown nature,was identified.RT-PCR and q RT-PCR validation indicated that orf610a was expressed specifically in a sterile line.Ectopic expression of orf610a in yeast resulted in excessive accumulation of reactive oxygen species and reduction in ATP content,in addition to inhibition of cellular growth.Transgenic A.thaliana overexpressing orf610a fused with a mitochondrial targeting peptide displayed partial male sterility.Interaction between ORF610a and the nuclear-encoded protein RD22 indicated an association between ORF610a and pollen abortion.Positive feedback during transcriptional regulation between nuclear regulatory factors and the mt CMS gene may account for the male sterility of ZBA.
基金supported by grants from the Beijing Scholars Program (BSP041)Youth Research Fund (QNJJ202128) of the Beijing Academy of Agriculture and Forestry SciencesScience and Technology Xing Meng Action Focus Project of Inner Mongolia Autonomous Region (KJXM2020001)。
文摘Moderate stimuli in mitochondria improve wideranging stress adaptability in animals, but whether mitochondria play similar roles in plants is largely unknown. Here, we report the enhanced stress adaptability of S-type cytoplasmic male sterility(CMS-S) maize and its association with mild expression of sterilizing gene ORF355. A CMS-S maize line exhibited superior growth potential and higher yield than those of the near-isogenic N-type line in saline fields. Moderate expression of ORF355 induced the accumulation of reactive oxygen species and activated the cellular antioxidative defense system. This adaptive response was mediated by elevation of the nicotinamide adenine dinucleotide concentration and associated metabolic homeostasis. Metabolome analysis revealed broad metabolic changes in CMS-S lines, even in the absence of salinity stress. Metabolic products associated with amino acid metabolism and galactose metabolism were substantially changed, which underpinned the alteration of the antioxidative defense system in CMS-S plants. The results reveal the ORF355-mediated superior stress adaptability in CMS-S maize and might provide an important route to developing salt-tolerant maize varieties.
基金supported by grants from the National Key Research and Development Programs of China(2016YFD0101803 and 2017YFD0300301).
文摘Coordination between mitochondria and the nucleus is crucial for fertility determination in plants with cytoplasmic male sterility(CMS).Using yeast one-hybrid screening,we identified a transcription factor,ZmDREB1.7,that is highly expressed in sterile microspores at the large vacuole stage and activates the expression of mitochondria-encoded CMS gene orf355.Δpro,a weak allele of ZmDREB1.7 with the loss of a key unfolded protein response(UPR)motif in the promoter,partially restores male fertility of CMS-S maize.ZmDREB1.7 expression increases rapidly in response to antimycin A treatment,but this response is attenuated in theΔpro allele.Furthermore,we found that expression of orf355 in mitochondria activates mitochondrial retrograde signaling,which in turn induces ZmDREB1.7 expression.Taken together,these findings demonstrate that positive-feedback transcriptional regulation between a nuclear regulator and a mitochondrial CMS gene determines male sterility in maize,providing new insights into nucleus-mitochondria communication in plants.
基金supported by the National Natural Science Foundation of China(31971893 and 31571745).
文摘Cytoplasmic male sterility(CMS)is a powerful tool for the exploitation of hybrid heterosis and the study of signaling and interactions between the nucleus and the cytoplasm.C-type CMS(CMS-C)in maize has long been used in hybrid seed production,but the underlying sterility factor and its mechanism of action remain unclear.In this study,we demonstrate that the mitochondrial gene atp6c confers male sterility in CMS-C maize.The ATP6C protein shows stronger interactions with ATP8 and ATP9 than ATP6 during the assembly of F1F0-ATP synthase(F-type ATP synthase,ATPase),thereby reducing the quantity and activity of assem-bled F_(1)F_(o)-ATP synthase.By contrast,the quantity and activity of the F1'component are increased in CMS-C lines.Reduced F1F0-ATP synthase activity causes accumulation of excess protons in the inner membrane space of the mitochondria,triggering a burst of reactive oxygen species(ROS),premature programmed cell death of the tapetai cells,and pollen abortion.Collectively,our study identifies a chimeric mitochondrial gene(ATP6C)that causes CMS in maize and documents the contribution of ATP6C to F1F0-ATP synthase assembly,thereby providing novel insights into the molecular mechanisms of male sterility in plants.
基金supported by the National Natural Science Foundation of China (31571745 and 31971893)the Key Technology Research and Development Program of Henan Province (202102110164 and 212102110061)+1 种基金the Zhengzhou Major Science and Technology Innovation Project (188PCXZX803)the Open Funds of the State Key Laboratory of Crop Genetics and Germplasm Enhancement (ZW202001)。
文摘Normal microsporogenesis is determined by both nuclear and mitochondrial genes. In maize C-type cytoplasmic male sterility, it is unclear how the development of meiocytes and microspores is affected by the mitochondrial sterility gene and the nuclear restorer gene. In this study, we sequenced the transcriptomes of single meiocytes(tetrad stage) and early mononucleate microspores from sterile and restorer lines. The numbers of expressed genes varied in individual cells and fewer than half of the expressed genes were common to the same cell types. Four comparisons revealed 3379 differentially expressed genes(DEGs), with 277 putatively associated with mitochondria, 226 encoding transcription factors,and 467 possibly targeted by RF4. KEGG analysis indicated that the DEGs in the two lines at the tetrad stage were involved predominantly in carbon metabolism and in amino acid biosynthesis and metabolism, whereas the DEGs during the transition from the tetrad stage to the early mononucleate stage were associated mostly with regulation of protein metabolism, fatty acid metabolism, and anatomical structure morphogenesis. Thus, meiocyte and microspore development was affected by the surrounding cells and the restorer gene, and the restorer gene helped restore the redox homeostasis of microspores and the normal cellular reconstruction during the transition.
基金This research was supported by the National Key Technology Research and Development Program of China(2016YFD0102102)the Sichuan Innovation Ability Rising Government Financial Project,China(2016ZYPZ-001)the Sichuan Rice Breeding Project for Tackling Key Problems,China(2016NYZ0028-1-1).
文摘The integrated use of molecular marker-assisted selection (MAS) and anther culture has potential to significantly increase efficiency in plant breeding;however, reports on this kind of practical use are very limited. In the present study, we report the development of cytoplasmic male sterile (CMS) maintainers with aroma, disease resistance and red-brown hulls, as an example of integration of MAS and anther culture in rice breeding. A high-resolution melting (HRM)-based functional molecular marker was developed for the red-brown hull trait caused by a unique mutation (rbh1) in OsCAD2. Functional molecular markers for genes of rice blast resistance (Pi2), aroma (fgr) and red-brown hull (rbh1) were used for precise genotyping of individual plants in the BC1 and BC2F2 populations derived from a cross between CMS maintainers Huaxiang B (pi2–/rbh1–/fgr–) and Rong 3B (Pi2+/RBH1+/Fgr+). A total of 89 doubled haploid (DH) lines were generated from selected BC2F2 plants (Pi2+/rbh1–/fgr–) by anther culture. Seven DH lines were subsequently selected as the potential new CMS maintainers based on their overall performance and high resistance to blast. Our study demonstrated that integration of MAS and anther culture significantly accelerated the development of CMS maintainers with multiple stacked genes.
基金the National Key Research and Development Program of China(Grant No.2016YFD0101503)the Key Research and Development Program of Anhui Province(Grant No.202004a06020034)+1 种基金the Major Science and Technology Project of Anhui Province(Grant No.18030701178)the Program on Industrial Technology System of National Soybean(Grant No.CARS-04-PS07)。
文摘Cytoplasmic male sterility(CMS)-restorer system is a useful tool to exploit heterosis in soybean.The major restorer gene for the M-type CMS is known as Rf-m,located in the 162.4-kb region on chromosome 16.Sequence analysis has revealed that the Rf-m locus in Glycine max consists of seven penta tricopeptide repeat(GmPPR)genes.The deduced amino acid sequences contain 8 to 14 PPR motifs,and a phylogenetic analysis grouped these GmPPR proteins into two PPR subfamilies:Glyma.16G161800 belongs to the PLS subfamily,and the P subfamily consists.of Glyma.16G161900,Glyma 16G162000,Glyma.16G162100,Glyma.16G162700,Glyma.16G162800,and Gly-ma 16G163100.The phylogenetic analysis of seven GmPPR proteins and 27 other plant PPR proteins also showed that proteins in the same subfamilies cluster together.Comparative sequence analysis was conducted using the seven Rf-m candidate GmPPR genes from the sterile line W931A,the maintainer line W931B,and the restorer line WR016,the result showed that Glyma 16G161900 had higher polymorphism than the other candidate genes.Based on real-time quantitative RT-PCR data,all seven GmPPR genes were differentially expressed but showed constitutive expression in roots,stems,leaves,and pollen grains.Additionally,the expression level of Gly-ma 16G161900 in the sterile line W931 A was significantly higher in all tissues than in the restorer line WR016.Taken together,these results suggest that Glyma 16G161900 is the most likely candidate for the restorer gene Rf-m.This study is the first report and analysis of candidate fertility restorer(Rf)genes encoding PPR proteins in soybean.
基金Supported by Funding of Utilization of Heterosis and Breeding of New Variety in Brassicaceous Vegetable(2012BAD02B01)
文摘Isolation of high-quality mitochondrial DNA(mtDNA) is an important premise for researching molecular mechanisms in cytoplasmic male sterility of cabbage(Brassica oleracea L.var.capitata). An efficient protocol for separation and purification of mitochondria and extraction of mitochondrial DNA(mtDNA) from etiolated tissues of cabbage was developed. We took a method combined mannitol density gradient with differential centrifugation, selected appropriate rotational speed, extended DNase I treating time and changed mitochondria cracking condition. The results showed that the extracted mitochondria in this protocol had complete structure, appeared to ellipsoid and had not been contaminated with other impurities under the Jannus Green B staining. The isolated mitochondrial DNA had high purity and yield through detecting the optical density, nuclear specific primer PCR and agarose gel electrophoresis. The results indicated that mitochondrial DNA extracted by this protocol had high quality and enabled to be used in futher genetic studies.
基金supported by grants from the National Natural Science Foundation of China(31788103)the Chinese Academy of Sciences(XDB27010301).
文摘The male sterility(MS)line is a prerequisite for efficient production of hybrid seeds in rice,a self-pollinating species.MS line breeding is pivotal for hybrid rice improvement.Understanding the historical breeding tra-jectory will help to improve hybrid rice breeding strategies.Maternally inherited cytoplasm is an appro-priate tool for phylogenetic reconstruction and pedigree tracing in rice hybrids.In this study,we analyzed the cytoplasmic genomes of 1495 elite hybrid rice varieties and identified five major types of cytoplasm,which correspond to different hybrid production systems.As the cytoplasm donors for hybrids,461 MS lines were also divided into five major types based on cytoplasmic and nuclear genomic architecture.Spe-cific core accessions cooperating with different fertility-associated genes drove the sequence divergence of MS lines.Dozens to hundreds of convergent and divergent selective sweeps spanning several agro-nomic trait-associated genes were identified among different types of MS lines.We further analyzed the cross patterns between different types of MS lines and their corresponding restorers.This study system-atically analyzed the cytoplasmic genomes of rice hybrids revealed their relationships with nuclear ge-nomes of MS lines,and illustrated the trajectory of hybrid rice breeding and the strategies for breeding different types of MS lines providing new insights for future improvement of hybrid rice.
基金supported by Heilongjiang Provincial Natural Science Foundation of China(Grant No.YQ2022C012)China Postdoctoral Science Foundation(Grant No.2022MD713728)+1 种基金Heilongjiang Provincial Postdoctoral Fund(Grant No.LBHZ21046)the Open Project of Key Laboratory of Biology and Genetic Improvement of Horticultural Crops(Northeast Region),Ministry of Agriculture and Rural Affairs,and National Key Research and Development Program of China(Grant No.2023YFD1201501).
文摘Ogura cytoplasmic male sterility(Ogura CMS)is extensively applied in hybrid seed production in cruciferous crops.However,the posttranscriptional molecular basis of Ogura CMS in cruciferous crops remains elusive.Here,a data-independent acquisition-based proteomic approach coupled with a parallel reaction monitoring-based targeted proteomic assay was used to analyze the proteome dynamics of Ogura CMS cabbage line RM and its maintainer line RF during floral bud development to obtain insights into the mechanism underlying Ogura CMS in cruciferous crops.A total of 9162 proteins corresponding to 61464 peptides were identified in RM and RF floral buds.The proteomic fluctuation of RM was weaker than that of RF.Differences in protein expression between RM and RF gradually enlarged with floral bud development.Fifteen continually up-regulated and eight continually down-regulated proteins were found in RM relative to RF throughout floral bud development.Differentially expressed proteins between RM and RF during floral bud development were implicated in the endoplasmic reticulum(ER)-associated protein processing pathway,in which most of them exhibited down-regulated expression in RM.These data suggest that ER-associated protein processing may be involved in pollen abortion in Ogura CMS cabbage by inhibiting the expression of critical factors.Our findings not only deepen the understanding of the molecular mechanisms of Ogura CMS in cruciferous crops but also provide better guidance for applying Ogura CMS in the hybrid breeding of cruciferous crops.
