Somatic embryogenesis of Fraxinus mandshurica has the problems of low somatic embryo(SE) yield,unsynchronized SE development,and a high percentage of deformed SEs.We aimed to improve F.mandshurica SE production by syn...Somatic embryogenesis of Fraxinus mandshurica has the problems of low somatic embryo(SE) yield,unsynchronized SE development,and a high percentage of deformed SEs.We aimed to improve F.mandshurica SE production by synchronizing SE development,improving SE quality,and inducing root formation to obtain complete regenerated plants.Cotyledons of immature zygotic embryos of F.mandshurica were induced to form callus and then SEs.The SE induction percentage from explants differed among 32 mother trees,and the one with the highest SE induction percentage(29.8%) was used for further experiments.The highest callus induction percentage was94.2% on 1/2-strength Murashige and Skoog medium(MS1/2)supplemented with 0.15 mg·L^(-1) naphthalene acetic acid.The highest callus proliferation coefficient(240.5) was obtained on McCown's Woody Plant Medium containing 0.1 mg·L^(-1)6-benzyl adenine and 0.15 mg·L^(-1) 2,4-dichlorophenoxyacetic acid.The highest number of SEs(1020.5 g~(-1) fresh weight) was obtained on MS1/2 medium supplemented with1 mg·L^(-1) 6-benzyladenine.The highest number of cotyledon embryos(397/g fresh weight) was obtained by incubating materials on medium containing 1 mg·L^(-1) abscisic acid and then applying a drying treatment.The cotyledon embryos were milky white,uniformly sized(average length 4.7 mm),and 80% of them were normal.The SE rooting percentage on 1/2 MS medium containing 0.01 mg·L^(-1) NAA was 37.5%.Overall,the germination percentage of SEs was 26.4%,and complete regenerated plants were obtained after transplanting and acclimation.These results provide more possibilities for the preservation and breeding of F.mandshurica.展开更多
Determining the influences of environmental factors on seedling growth and leaf color may improve our understanding of the chromogenic mechanisms in leaves and the biosynthesis of anthocyanin. To determine the optimum...Determining the influences of environmental factors on seedling growth and leaf color may improve our understanding of the chromogenic mechanisms in leaves and the biosynthesis of anthocyanin. To determine the optimum conditions for Betula ‘Royal Frost' seedling cultivation, we assessed seedling growth and anthocyanin content under natural conditions. The temperature had a significant influence on height but not on radial growth.Between June and September, the anthocyanin content was significantly positively correlated with light intensity and air moisture and significantly negatively correlated with temperature. In single-factor experiments to determine the variation in anthocyanin content in the greenhouse, anthocyanin content increased with increasing soil water content and at first increased, then decreased with increasing p H of the irrigation water and temperature, but decreased with increasing light intensity. K_2SO_4 can increase the anthocyanin content. So to maintain leaf color, seedlings should be cultivated with< 200 lmol m^(-2) s^(-1) light intensity at 20 °C and soil moisture content between 60 and 90%, and the irrigation water should be neutral or alkaline. Additionally, spraying with K_2SO_4 can be beneficial.展开更多
The induction and proliferation of embryogenic callus are key steps for large-scale propagation of somatic embryogenesis pathway and long-term preservation of coniferous germplasm.Callus can be induced from immature e...The induction and proliferation of embryogenic callus are key steps for large-scale propagation of somatic embryogenesis pathway and long-term preservation of coniferous germplasm.Callus can be induced from immature embryos of Korean pine(Pinus koraiensis Sieb.et Zucc.;Pinaceae)as explants,but there are problems,such as low proliferation efficiency,loss of embryogenicity,poor vigor;thus,best conditions for proliferation and culture of immature embryos of Korean pine are not yet clear.To solve the problems with somatic embryogenesis of Korean pine and determine the best culture conditions for callus induction and proliferation,we varied hormone concentration,subculture cycle of proliferation and other plant growth regulators combinations in media to induce callus formation by megagametophytes of three Korean pine families at different developmental stages,then analyzed the effects on embryogenic callus retention and cell proliferation using a quadratic regression orthogonal rotation design.The results showed that the family origin and collection date of explants significantly affected callus induction(induction rate reached 1.67%).Embryogenic maintenance and callus proliferation were best on DCR medium supplemented with 0.25 mg L^(−1)6-benzyl adenine,1 mg L^(−1)naphthaleneacetic acid,30 g L^(−1)sucrose,500 mg L^(−1),L-glutamine,500 mg L^(−1)casein hydrolysis and 6.5 g L^(−1)agar.In addition,the combination of 2,4-dichlorophenoxyacetic acid+6-benzyl adenine also had a better proliferative effect on callus.The effects of different combinations of growth regulators on callus proliferation efficiency were significantly different.Transfer to new medium every 13–15 days not only maintained robust callus vigor,but also yielded a larger proliferation coefficient.The techniques and conditions for embryogenic callus induction and proliferation of Korean determined here will serve as a foundation for establishing a large-scale system for somatic embryogenesis and propagation of Korean pine.展开更多
We examined how reactive oxygen species, in the form of hydrogen peroxide (H2O2), affect osmotic stress–induced programmed cell death during somatic embryogenesis from cotyledon explants of Manchurian ash (Fraxinus m...We examined how reactive oxygen species, in the form of hydrogen peroxide (H2O2), affect osmotic stress–induced programmed cell death during somatic embryogenesis from cotyledon explants of Manchurian ash (Fraxinus mandshurica Rupr.). We found that substantial osmotic stress was essential for Manchurian ash somatic cells to obtain embryogenic competence. The explant cells displayed hallmarks of programmed cell death, chromatin condensation, and DNA fragmentation to oligonucleotides during somatic embryogenesis. Increasing concentrations of plant growth regulators and sucrose in the medium increased osmotic stress thereby inducing H2O2 accumulation in the explant cells. We found that H2O2 concentration was significantly decreased in explant cells when the induction medium was modified, i.e., when reducing the concentration of sucrose, which reduces the osmotic pressure of the medium, or by withdrawing plant growth regulators at mid-culture. These treatments also decreased the proportion of explant cells undergoing programmed cell death. Accordingly, a decreased rate of somatic embryo induction was observed. These results show that PCD occurred during tissue browning and death of some explant cells during somatic embryogenesis in F. mandshurica. The ROS contributed to PCD in abiotic stress stimulated F. mandshurica cells.展开更多
The effects of an exogenous nitric oxide donor(sodium nitroprusside, SNP), a NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxode(PTIO) and carboxy-PTIO potassium salt(c PTIO) on the embryo germination ...The effects of an exogenous nitric oxide donor(sodium nitroprusside, SNP), a NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxode(PTIO) and carboxy-PTIO potassium salt(c PTIO) on the embryo germination of Sorbus pohuashanensis were studied in a petri dish test. SNP at 0.5–5 mmol L-1 increased germination percentage, mean time to germination, germination index and germination energy compared with the control to different degrees. Treatment with 2 mmol L-1 SNP improved germination most significantly; embryo germination percentage for mother tree 1(91.11%) and mother tree 2(64.44%) were much higher than the control. In addition,excessive SNP levels did not enhance embryo germination.Combined treatment with SNP and an NO scavenger delayed embryo germination. Treatment with c PTIO inhibited embryo germination; germination percentage was 42.22% and was lower than that of the control. These results show that low concentrations of exogenous NO can enhance the embryo germination of S. pohuashanensis,providing a simple, effective way for promoting germination of S. pohuashanensis.展开更多
Acer mono Maxim. is one of the major components of cool temperate forests in Japan. Some of its many varieties are distributed sympatrically. Because of its great variability, the intraspecific taxonomy and nomenclatu...Acer mono Maxim. is one of the major components of cool temperate forests in Japan. Some of its many varieties are distributed sympatrically. Because of its great variability, the intraspecific taxonomy and nomenclature of the species are controversial. To understand the genetic relationships among these varieties and whether hybridization or introgression occurred among the sympatric varieties, we studied the genetic relationships among sympatric varieties of A. mono in the Chichibu Mountains(A. mono var. ambiguum, A. mono var. connivens, A. mono var. marmoratum) and Central Hokkaido(A. mono var.mayrii and A. mono var. glabrum) in Japan. Our results showed that varieties in Chichibu are genetically close,suggesting that hybridization or introgression might occur between these varieties, which could explain the higher genetic diversity of varieties in Chichibu than in Hokkaido.In contrast to the close relationships between the varieties in Chichibu, varieties in Hokkaido seemed relatively separated from each other; indeed, there may be reproductive isolation between the two varieties. The results provide new insight for the taxonomy of the varieties of A. mono,especially the sympatric varieties, in Japan.展开更多
Programmed cell death occurs in browning explants of Fraxinus mandshurica during somatic embryogenesis, but the underlying mechanism is unclear. In this study, single cotyledons of zygotic embryos of F. mandshurica we...Programmed cell death occurs in browning explants of Fraxinus mandshurica during somatic embryogenesis, but the underlying mechanism is unclear. In this study, single cotyledons of zygotic embryos of F. mandshurica were used as explants. Mitochondrial structure and function, caspase-3-like protease activity, hydrogen peroxide metabolism, and nitric oxide accumulation induced by high concentrations of sucrose and plant growth regulators were studied. The results show that plant growth regulators induced somatic embryogenesis and also promoted explant browning. High sucrose concentrations had similar effects. High concentrations of sucrose and plant growth regulators led to the accumulation of hydrogen peroxide and nitric oxide which induced changes in mitochondrial structure and function such as modifications in mitochondrial morphology, increased membrane permeability, decreased membrane potential, and the release of cytochrome c into the cytoplasm. An increase in caspase-3-like protease activity triggered programmed cell death in some browning explant cells. During somatic embryogenesis there were increased activities of superoxide dismutase, peroxidase, and catalase, which are associated with hydrogen peroxide metabolism and jointly maintain reactive oxygen species levels. Intracellular nitric oxide synthase and nitrate reductase activities were not significantly correlated with nitric oxide content. Instead, intracellular nitric oxide may be derived from non-enzymatic reactions. Our results indicate that hydrogen peroxide and nitric oxide may function as signals, playing key roles in somatic embryogenesis and programmed cell death of explant cells of F. mandshurica. The interaction between nitric oxide and reactive oxygen species determines the occurrence of programmed cell death in explant cells;somatic embryogenesis and programmed cell death are positively regulated by hydrogen peroxide. However, the regulation of nitric oxide is complex.展开更多
The breaking of dormancy mediated by reac-tive nitrogen species(RNS)is related to the accumulation of reactive oxygen species(ROS)in germinating embryos but the underlying mechanism is unclear.The objectives of this s...The breaking of dormancy mediated by reac-tive nitrogen species(RNS)is related to the accumulation of reactive oxygen species(ROS)in germinating embryos but the underlying mechanism is unclear.The objectives of this study were:(1)to explore the relationship between RNS-mediated dormancy release and ROS accumulation in germinating embryos of Sorbus pohuashanensis;and,(2)to investigate the relationships among germination time,ROS metabolism,and endogenous hormone synthesis.We studied the effects of exogenous nitric oxide(NO)donor sodium nitroprusside(SNP),the NO scavenger(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide(PTIO),abscisic acid(ABA),the exogenous ethylene donor ethrel,and the ethylene receptor inhibitor 2,5-norbornadien(NBD)on embryo germination and seedling growth.Embryos were released from dormancy by pretreatment with NO or ethylene, which was related to increased ethylene biosyn-thesis and decreased ABA levels. Breaking of dormancy by SNP was related to increased levels of ethylene, hydrogen peroxide, and glutathione, increased activities of superoxide dismutase and glutathione peroxidase, and decreased levels of ABA, superoxide anions, and malondialdehyde. These effects of nitric oxide were especially significant in seedling hypocotyls and radicles. These results demonstrate that NO can break S. pohuashanensis embryo dormancy by induc-ing ethylene biosynthesis, and that this signalling pathway is closely related to ROS accumulation and the antioxidant defence response.展开更多
Osmotic stress promotes somatic embryogenesis of Fraxinus mandshurica,which leads to accumulation of reactive oxygen species(ROS).The single pieces of cotyledons of F.mandshurica were used as explants to induce somati...Osmotic stress promotes somatic embryogenesis of Fraxinus mandshurica,which leads to accumulation of reactive oxygen species(ROS).The single pieces of cotyledons of F.mandshurica were used as explants to induce somatic embryogenesis in osmotic-stress medium.Furthermore,the hydrogen peroxide H_(2)O_(2) content of explanted cells was varied by adding exogenous H_(2)O_(2) or catalase solution to assess the effects of the exogenous H_(2)O_(2)on somatic embryogenesis,intracellular H_(2)O_(2)accumulation,and the relationship between signaling mediated by ROS or reactive nitrogen species.The results revealed that exogenous H_(2)O_(2)(100?300μmol L^(–1))increased the number of somatic embryos.On 60th day of exogenous H_(2)O_(2)(200μmol L^(–1))treatment,the number of somatic embryos of explants treated,which was 136.54%,was higher than the control.Moreover,exogenous H_(2)O_(2)(100μmol L^(–1))significantly increased the intracellular H_(2)O_(2)content and enhanced the activities of superoxidase dismutase and peroxidase.Finally,exogenous H_(2)O_(2)(100μmol L^(–1))activated the intracellular non-enzymatic pathway for nitric oxide(NO)synthesis.The somatic embryogenesis in broadleaf trees increases with the change of endogenic ROS content,and depends on the upregulation of antioxidant enzymes.Both H_(2)O_(2)and NO,as signaling molecules,were found to be involved in the process of somatic embryogenesis in broadleaf trees.In the process of exogenous H_(2)O_(2)promoting somatic embryogenesis,NO synthesis depended on non-enzymatic reactions.These results provide a scientific basis for resolving the mechanism by which ROS levels are regulated during somatic embryogenesis of broadleaf trees and establish a reasonable and efficient technology system for regulating somatic embryogenesis of trees.展开更多
Correction to:J.For.Res.https://doi.org/10.1007/s11676-021-01392-y In this article Ling Yang should have been denoted as a corresponding author.The original article has been corrected.Publisher’s Note Springer Nature...Correction to:J.For.Res.https://doi.org/10.1007/s11676-021-01392-y In this article Ling Yang should have been denoted as a corresponding author.The original article has been corrected.Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affi liations.展开更多
Korean pine is an important afforestation tree species in Northeast China,which has a high ecological and economic value.Although regeneration of somatic embryogenesis using immature zygotic embryos of Korean pine as ...Korean pine is an important afforestation tree species in Northeast China,which has a high ecological and economic value.Although regeneration of somatic embryogenesis using immature zygotic embryos of Korean pine as explants has been successful,it cannot be applied to automation and large-scale production.Therefore,we urgently need a method that can increase the output of somatic embryos(SEs)to meet the needs of large-scale production.We used Korean pine 1-1 and 1-100 cell lines as research materials to evaluate the effects of inoculum-density,culture time,orbiting speed,vessel volume,plant growth regulator(PGR)concentration,and carbon source on the proliferation of embryogenic tissue(ET).The somatic embryogenesis ability of ET cultured in different liquid suspension media was also evaluated.We found that during liquid suspension culture of Korean pine ET,the sedimented cell volume(SCV),fresh weight(FW)and dry weight(DW)were affected by inoculumdensity,culture time,orbiting speed,2,4-D concentration,6-BA concentration and carbon source type.Fourty mg⋅mL^(−1)ET were transferred to a 200 mL Erlenmeyer flask containing 20 mL liquid medium,and cultured at 100 rpm/min for 14 days to obtain the maximum proliferation.In addition,we also found that SCV,FW and DW were higher when PGRs were reduced in the liquid suspension medium.The substitution of maltose for sucrose resulted in slow growth of cultures and limited SE yield(13 SEs g^(−1)FW).Although culture proliferation was high at 50 rpm,SE yield was inhibited by 48%compared with 100 rpm(50 rpm=33 SEs g^(−1)FW;100 rpm/min=70 SEs g^(−1)FW).Cultivation in low-concentration PGR(1.15μM⋅L^(−1)2,4-D,0.25μM⋅L^(−1)6-BA)and sucrose liquid medium at 100 rpm/min(80 SEs g^(−1)FW)could not only promote culture proliferation but also increase SE yield.The determination of the suspension culture scheme of Korean pine ET provides a reference for further expansion to bioreactor culture in the future and lays a foundation for the automation and scale of somatic embryogenesis of Korean pine.展开更多
基金supported by the Fundamental Research Funds for the Central Universities of China(2572018BW02)the National Natural Science Foundation of China (31400535 and 31570596)+1 种基金the Innovation Project of State Key Laboratory of Tree Genetics and Breeding (2016C01)the National Key R&D Program of China (2017YFD0600600)。
文摘Somatic embryogenesis of Fraxinus mandshurica has the problems of low somatic embryo(SE) yield,unsynchronized SE development,and a high percentage of deformed SEs.We aimed to improve F.mandshurica SE production by synchronizing SE development,improving SE quality,and inducing root formation to obtain complete regenerated plants.Cotyledons of immature zygotic embryos of F.mandshurica were induced to form callus and then SEs.The SE induction percentage from explants differed among 32 mother trees,and the one with the highest SE induction percentage(29.8%) was used for further experiments.The highest callus induction percentage was94.2% on 1/2-strength Murashige and Skoog medium(MS1/2)supplemented with 0.15 mg·L^(-1) naphthalene acetic acid.The highest callus proliferation coefficient(240.5) was obtained on McCown's Woody Plant Medium containing 0.1 mg·L^(-1)6-benzyl adenine and 0.15 mg·L^(-1) 2,4-dichlorophenoxyacetic acid.The highest number of SEs(1020.5 g~(-1) fresh weight) was obtained on MS1/2 medium supplemented with1 mg·L^(-1) 6-benzyladenine.The highest number of cotyledon embryos(397/g fresh weight) was obtained by incubating materials on medium containing 1 mg·L^(-1) abscisic acid and then applying a drying treatment.The cotyledon embryos were milky white,uniformly sized(average length 4.7 mm),and 80% of them were normal.The SE rooting percentage on 1/2 MS medium containing 0.01 mg·L^(-1) NAA was 37.5%.Overall,the germination percentage of SEs was 26.4%,and complete regenerated plants were obtained after transplanting and acclimation.These results provide more possibilities for the preservation and breeding of F.mandshurica.
基金supported by the Postdoctoral Science Foundation of Heilongjiang Province of China(LBHQ14003)the National Natural Science Foundation of China(No.31400535)
文摘Determining the influences of environmental factors on seedling growth and leaf color may improve our understanding of the chromogenic mechanisms in leaves and the biosynthesis of anthocyanin. To determine the optimum conditions for Betula ‘Royal Frost' seedling cultivation, we assessed seedling growth and anthocyanin content under natural conditions. The temperature had a significant influence on height but not on radial growth.Between June and September, the anthocyanin content was significantly positively correlated with light intensity and air moisture and significantly negatively correlated with temperature. In single-factor experiments to determine the variation in anthocyanin content in the greenhouse, anthocyanin content increased with increasing soil water content and at first increased, then decreased with increasing p H of the irrigation water and temperature, but decreased with increasing light intensity. K_2SO_4 can increase the anthocyanin content. So to maintain leaf color, seedlings should be cultivated with< 200 lmol m^(-2) s^(-1) light intensity at 20 °C and soil moisture content between 60 and 90%, and the irrigation water should be neutral or alkaline. Additionally, spraying with K_2SO_4 can be beneficial.
基金supported by the National Key R&D Program of China(2017YFD0600600).
文摘The induction and proliferation of embryogenic callus are key steps for large-scale propagation of somatic embryogenesis pathway and long-term preservation of coniferous germplasm.Callus can be induced from immature embryos of Korean pine(Pinus koraiensis Sieb.et Zucc.;Pinaceae)as explants,but there are problems,such as low proliferation efficiency,loss of embryogenicity,poor vigor;thus,best conditions for proliferation and culture of immature embryos of Korean pine are not yet clear.To solve the problems with somatic embryogenesis of Korean pine and determine the best culture conditions for callus induction and proliferation,we varied hormone concentration,subculture cycle of proliferation and other plant growth regulators combinations in media to induce callus formation by megagametophytes of three Korean pine families at different developmental stages,then analyzed the effects on embryogenic callus retention and cell proliferation using a quadratic regression orthogonal rotation design.The results showed that the family origin and collection date of explants significantly affected callus induction(induction rate reached 1.67%).Embryogenic maintenance and callus proliferation were best on DCR medium supplemented with 0.25 mg L^(−1)6-benzyl adenine,1 mg L^(−1)naphthaleneacetic acid,30 g L^(−1)sucrose,500 mg L^(−1),L-glutamine,500 mg L^(−1)casein hydrolysis and 6.5 g L^(−1)agar.In addition,the combination of 2,4-dichlorophenoxyacetic acid+6-benzyl adenine also had a better proliferative effect on callus.The effects of different combinations of growth regulators on callus proliferation efficiency were significantly different.Transfer to new medium every 13–15 days not only maintained robust callus vigor,but also yielded a larger proliferation coefficient.The techniques and conditions for embryogenic callus induction and proliferation of Korean determined here will serve as a foundation for establishing a large-scale system for somatic embryogenesis and propagation of Korean pine.
基金supported by the National Key R&D Program of China(2017YFD0600600)the National Natural Science Foundation of China(Nos.31400535 and 31570596)+2 种基金the National Basic Science Personnel Training Fund of China(No.J1210053)the Innovation Project of State Key Laboratory of Tree Genetics and Breeding(Northeast Forestry University,No.2016C01)the Fundamental Research Funds for the Central Universities(Nos.2572014CA13 and 2572018BW02)
文摘We examined how reactive oxygen species, in the form of hydrogen peroxide (H2O2), affect osmotic stress–induced programmed cell death during somatic embryogenesis from cotyledon explants of Manchurian ash (Fraxinus mandshurica Rupr.). We found that substantial osmotic stress was essential for Manchurian ash somatic cells to obtain embryogenic competence. The explant cells displayed hallmarks of programmed cell death, chromatin condensation, and DNA fragmentation to oligonucleotides during somatic embryogenesis. Increasing concentrations of plant growth regulators and sucrose in the medium increased osmotic stress thereby inducing H2O2 accumulation in the explant cells. We found that H2O2 concentration was significantly decreased in explant cells when the induction medium was modified, i.e., when reducing the concentration of sucrose, which reduces the osmotic pressure of the medium, or by withdrawing plant growth regulators at mid-culture. These treatments also decreased the proportion of explant cells undergoing programmed cell death. Accordingly, a decreased rate of somatic embryo induction was observed. These results show that PCD occurred during tissue browning and death of some explant cells during somatic embryogenesis in F. mandshurica. The ROS contributed to PCD in abiotic stress stimulated F. mandshurica cells.
基金supported by the Science Foundation of Heilongjiang Province of China(C201407)the National Natural Science Foundation of China(No.31400535)
文摘The effects of an exogenous nitric oxide donor(sodium nitroprusside, SNP), a NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxode(PTIO) and carboxy-PTIO potassium salt(c PTIO) on the embryo germination of Sorbus pohuashanensis were studied in a petri dish test. SNP at 0.5–5 mmol L-1 increased germination percentage, mean time to germination, germination index and germination energy compared with the control to different degrees. Treatment with 2 mmol L-1 SNP improved germination most significantly; embryo germination percentage for mother tree 1(91.11%) and mother tree 2(64.44%) were much higher than the control. In addition,excessive SNP levels did not enhance embryo germination.Combined treatment with SNP and an NO scavenger delayed embryo germination. Treatment with c PTIO inhibited embryo germination; germination percentage was 42.22% and was lower than that of the control. These results show that low concentrations of exogenous NO can enhance the embryo germination of S. pohuashanensis,providing a simple, effective way for promoting germination of S. pohuashanensis.
基金supported by National Postdoctoral Daily Fund of China and Heilongjiang Postdoctoral Fund(LBHZ13007)
文摘Acer mono Maxim. is one of the major components of cool temperate forests in Japan. Some of its many varieties are distributed sympatrically. Because of its great variability, the intraspecific taxonomy and nomenclature of the species are controversial. To understand the genetic relationships among these varieties and whether hybridization or introgression occurred among the sympatric varieties, we studied the genetic relationships among sympatric varieties of A. mono in the Chichibu Mountains(A. mono var. ambiguum, A. mono var. connivens, A. mono var. marmoratum) and Central Hokkaido(A. mono var.mayrii and A. mono var. glabrum) in Japan. Our results showed that varieties in Chichibu are genetically close,suggesting that hybridization or introgression might occur between these varieties, which could explain the higher genetic diversity of varieties in Chichibu than in Hokkaido.In contrast to the close relationships between the varieties in Chichibu, varieties in Hokkaido seemed relatively separated from each other; indeed, there may be reproductive isolation between the two varieties. The results provide new insight for the taxonomy of the varieties of A. mono,especially the sympatric varieties, in Japan.
基金This work was supported by the Fundamental Research Funds for the Central Universities(2572018BW02)the Innovation Project of State Key Laboratory of Tree Genetics and Breeding(2016C01)+1 种基金the National Key R&D Program of China(2017YFD0600600)the National Natural Science Foundation of China(31400535 and 31570596).
文摘Programmed cell death occurs in browning explants of Fraxinus mandshurica during somatic embryogenesis, but the underlying mechanism is unclear. In this study, single cotyledons of zygotic embryos of F. mandshurica were used as explants. Mitochondrial structure and function, caspase-3-like protease activity, hydrogen peroxide metabolism, and nitric oxide accumulation induced by high concentrations of sucrose and plant growth regulators were studied. The results show that plant growth regulators induced somatic embryogenesis and also promoted explant browning. High sucrose concentrations had similar effects. High concentrations of sucrose and plant growth regulators led to the accumulation of hydrogen peroxide and nitric oxide which induced changes in mitochondrial structure and function such as modifications in mitochondrial morphology, increased membrane permeability, decreased membrane potential, and the release of cytochrome c into the cytoplasm. An increase in caspase-3-like protease activity triggered programmed cell death in some browning explant cells. During somatic embryogenesis there were increased activities of superoxide dismutase, peroxidase, and catalase, which are associated with hydrogen peroxide metabolism and jointly maintain reactive oxygen species levels. Intracellular nitric oxide synthase and nitrate reductase activities were not significantly correlated with nitric oxide content. Instead, intracellular nitric oxide may be derived from non-enzymatic reactions. Our results indicate that hydrogen peroxide and nitric oxide may function as signals, playing key roles in somatic embryogenesis and programmed cell death of explant cells of F. mandshurica. The interaction between nitric oxide and reactive oxygen species determines the occurrence of programmed cell death in explant cells;somatic embryogenesis and programmed cell death are positively regulated by hydrogen peroxide. However, the regulation of nitric oxide is complex.
基金provided by National Natural Science Foundation of China(No.32071757)National Natural Science Foundation of Heilongjiang Province of China(C201407).
文摘The breaking of dormancy mediated by reac-tive nitrogen species(RNS)is related to the accumulation of reactive oxygen species(ROS)in germinating embryos but the underlying mechanism is unclear.The objectives of this study were:(1)to explore the relationship between RNS-mediated dormancy release and ROS accumulation in germinating embryos of Sorbus pohuashanensis;and,(2)to investigate the relationships among germination time,ROS metabolism,and endogenous hormone synthesis.We studied the effects of exogenous nitric oxide(NO)donor sodium nitroprusside(SNP),the NO scavenger(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide(PTIO),abscisic acid(ABA),the exogenous ethylene donor ethrel,and the ethylene receptor inhibitor 2,5-norbornadien(NBD)on embryo germination and seedling growth.Embryos were released from dormancy by pretreatment with NO or ethylene, which was related to increased ethylene biosyn-thesis and decreased ABA levels. Breaking of dormancy by SNP was related to increased levels of ethylene, hydrogen peroxide, and glutathione, increased activities of superoxide dismutase and glutathione peroxidase, and decreased levels of ABA, superoxide anions, and malondialdehyde. These effects of nitric oxide were especially significant in seedling hypocotyls and radicles. These results demonstrate that NO can break S. pohuashanensis embryo dormancy by induc-ing ethylene biosynthesis, and that this signalling pathway is closely related to ROS accumulation and the antioxidant defence response.
基金supported by the National Natural Science Foundation of China(31570596 and 31400535)the Fundamental Research Funds for the Central Universities(2572018BW02)+1 种基金the Innovation Project of State Key Laboratory of Tree Genetics and Breeding(Northeast Forestry University,2016C01)the National Key R&D Program of China(2017YFD0600600)。
文摘Osmotic stress promotes somatic embryogenesis of Fraxinus mandshurica,which leads to accumulation of reactive oxygen species(ROS).The single pieces of cotyledons of F.mandshurica were used as explants to induce somatic embryogenesis in osmotic-stress medium.Furthermore,the hydrogen peroxide H_(2)O_(2) content of explanted cells was varied by adding exogenous H_(2)O_(2) or catalase solution to assess the effects of the exogenous H_(2)O_(2)on somatic embryogenesis,intracellular H_(2)O_(2)accumulation,and the relationship between signaling mediated by ROS or reactive nitrogen species.The results revealed that exogenous H_(2)O_(2)(100?300μmol L^(–1))increased the number of somatic embryos.On 60th day of exogenous H_(2)O_(2)(200μmol L^(–1))treatment,the number of somatic embryos of explants treated,which was 136.54%,was higher than the control.Moreover,exogenous H_(2)O_(2)(100μmol L^(–1))significantly increased the intracellular H_(2)O_(2)content and enhanced the activities of superoxidase dismutase and peroxidase.Finally,exogenous H_(2)O_(2)(100μmol L^(–1))activated the intracellular non-enzymatic pathway for nitric oxide(NO)synthesis.The somatic embryogenesis in broadleaf trees increases with the change of endogenic ROS content,and depends on the upregulation of antioxidant enzymes.Both H_(2)O_(2)and NO,as signaling molecules,were found to be involved in the process of somatic embryogenesis in broadleaf trees.In the process of exogenous H_(2)O_(2)promoting somatic embryogenesis,NO synthesis depended on non-enzymatic reactions.These results provide a scientific basis for resolving the mechanism by which ROS levels are regulated during somatic embryogenesis of broadleaf trees and establish a reasonable and efficient technology system for regulating somatic embryogenesis of trees.
文摘Correction to:J.For.Res.https://doi.org/10.1007/s11676-021-01392-y In this article Ling Yang should have been denoted as a corresponding author.The original article has been corrected.Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affi liations.
基金This work was supported by the Fundamental Research Funds for the Central Universities(2572020AW15)the National Key R&D Program of China(2017YFD0600600).
文摘Korean pine is an important afforestation tree species in Northeast China,which has a high ecological and economic value.Although regeneration of somatic embryogenesis using immature zygotic embryos of Korean pine as explants has been successful,it cannot be applied to automation and large-scale production.Therefore,we urgently need a method that can increase the output of somatic embryos(SEs)to meet the needs of large-scale production.We used Korean pine 1-1 and 1-100 cell lines as research materials to evaluate the effects of inoculum-density,culture time,orbiting speed,vessel volume,plant growth regulator(PGR)concentration,and carbon source on the proliferation of embryogenic tissue(ET).The somatic embryogenesis ability of ET cultured in different liquid suspension media was also evaluated.We found that during liquid suspension culture of Korean pine ET,the sedimented cell volume(SCV),fresh weight(FW)and dry weight(DW)were affected by inoculumdensity,culture time,orbiting speed,2,4-D concentration,6-BA concentration and carbon source type.Fourty mg⋅mL^(−1)ET were transferred to a 200 mL Erlenmeyer flask containing 20 mL liquid medium,and cultured at 100 rpm/min for 14 days to obtain the maximum proliferation.In addition,we also found that SCV,FW and DW were higher when PGRs were reduced in the liquid suspension medium.The substitution of maltose for sucrose resulted in slow growth of cultures and limited SE yield(13 SEs g^(−1)FW).Although culture proliferation was high at 50 rpm,SE yield was inhibited by 48%compared with 100 rpm(50 rpm=33 SEs g^(−1)FW;100 rpm/min=70 SEs g^(−1)FW).Cultivation in low-concentration PGR(1.15μM⋅L^(−1)2,4-D,0.25μM⋅L^(−1)6-BA)and sucrose liquid medium at 100 rpm/min(80 SEs g^(−1)FW)could not only promote culture proliferation but also increase SE yield.The determination of the suspension culture scheme of Korean pine ET provides a reference for further expansion to bioreactor culture in the future and lays a foundation for the automation and scale of somatic embryogenesis of Korean pine.
