The thermal-elastic martensitic transformation from high-temperature Ni_(2)In-type hexagonal structure to low-temperature TiNiSi-type orthorhombic structure has been widely studied in MnMX(M=Ni or Co,and X=Ge or Si)al...The thermal-elastic martensitic transformation from high-temperature Ni_(2)In-type hexagonal structure to low-temperature TiNiSi-type orthorhombic structure has been widely studied in MnMX(M=Ni or Co,and X=Ge or Si)alloys.However,the answer to how the orthorhombic martensite nucleates and grows within the hexagonal parent is still unclear.In this work,the hexagonal-orthorhombic martensitic transformation in a Co and Ge co-substituted MnNiSi is investigated.One can find some orthorhombic laths embedded in the hexagonal parent at a temperature above the martensitic transformation start temperature(M_(s)).With the the sample cooing to M_(s),the laths turn broader,indicating that the martensitic transformation starts from these pre-existing orthorhombic laths.Microstructure observation suggests that these pre-existing orthorhombic laths do not originate from the hexagonal-orthorhombic martensitic transformation because of the difference between atomic occupations of doping elements in the hexagonal parent and those in the preexisting orthorhombic laths.The phenomenological crystallographic theory and experimental investigations prove that the pre-existing orthorhombic lath and generated orthorhombic martensite have the same crystallography relationship to the hexagonal parent.Therefore,the orthorhombic martensite can take these pre-existing laths as embryos and grow up.This work implies that the martensitic transformation in MnNiSi_(1-x)(CoNiGe)_(x) alloy is initiated by orthorhombic embryos.展开更多
In previous study we reported that pretreatment with plasmolysis enhanced somatic embryo formation in hypocotyls of Eleutherococcus senticosus.In the present study,the expression level of callose synthase gene in embr...In previous study we reported that pretreatment with plasmolysis enhanced somatic embryo formation in hypocotyls of Eleutherococcus senticosus.In the present study,the expression level of callose synthase gene in embryos of E.senticosus in response to 2,4-D,sucrose and mannitol treatments was analyzed by RT-PCR.The results show that plasmolysis pretreatment using sucrose and mannitol significantly promoted the expression of callose synthase gene.Also,the thicker cell walls of explant plasmolyzed compared with controls were observed during the somatic embryogenesis.We suggest that the callose may make the cells in epidermis separate from neighboring cells and then develop into embryogenic potential cells.展开更多
We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera U 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) ...We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera U 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) medium supplemented with 1.0 mg·L^-1 2,4-D and 0.5 mg·L^-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg·L^-1 6-BA and 2 mg·L^-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg·L^-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dynamic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic embryos, including typical epiderma, cotyledon primordium and vascular tissue.展开更多
With embryogenic callus of Larix olgensisis, we investigated the effects of inositol, glutamine, casein hydrolysate, carbohydrate, abscisic acid and silver nitrate concentration on the maturation of the somatic embryo...With embryogenic callus of Larix olgensisis, we investigated the effects of inositol, glutamine, casein hydrolysate, carbohydrate, abscisic acid and silver nitrate concentration on the maturation of the somatic embryo.Three dominant factors emerged, and we developed a response surface model based on the Box-Behnken design.We defined the optimal conditions for the maturation of somatic embryos. The contents of abscisic acid, silver nitrate, sucrose and casein hydrolysis significantly affected the amount of maturing embryos, but inositol, maltose and glutamine had no effect. By establishing a response surface model with multiple factors, we predicted that the optimal number of L. olgensis somatic embryos was 204 ± 4 gon basal medium, containing 18.28 mg Labscisic acid,5.46 mg Lsilver nitrate and 82.67 g Lsucrose. In the verification experiments, the addition of 20 mg Labscisic acid, 5 mg Lsilver nitrate and 80 g Lsucrose to BM yielded an average of 202.06 somatic embryos per gram. These results should guide large-scale breeding of L. olgensis.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
A description of a successful direct somatic embryogenesis induction from immature zygotic embryos of a camphor tree (Cinnamomum camphora L.) is presented. After a subculture of 2-3 years, embryogenic calli could be...A description of a successful direct somatic embryogenesis induction from immature zygotic embryos of a camphor tree (Cinnamomum camphora L.) is presented. After a subculture of 2-3 years, embryogenic calli could be derived from primary somatic embryos. Immature zygotic embryos were cultured on a Murashige and Skoog (MS) basal medium supplemented with a range of combinations of cytokinins (BA) and auxins (2,4-D or NAA) for somatic embryo induction. Primary somatic embryos could be induced directly in almost all PGR combinations. A positive effect of 2,4-D on somatic embryogenesis from immature zygotic embryos of camphor tree was obtained. BA at appropriate concentrations (〈 5 mg-L-1) had an effect similar to 2,4-D, whereas high concentrations (〉 5 mg·L^-1) of BA had the effect of restraining somatic embryo induction. NAA had a less positive effect on somatic embryogenesis than 2,4-D.展开更多
Pinus patula is a species commonly used for reforestation in Mexico.However,efficient methods for the mass production seedlings are required.Micropropagation particularly by somatic embryogenesis provides an option fo...Pinus patula is a species commonly used for reforestation in Mexico.However,efficient methods for the mass production seedlings are required.Micropropagation particularly by somatic embryogenesis provides an option for the rapid multiplication of high-quality,genetically improved material.This study induces somatic embryogenesis in this species using the thin cell layer(TCL)technique.Two sources of explants(complete immature embryos;lTCL segments from immature embryos)were evaluated.The efficiency of TCL from longitudinal sections[lTCL]and transverse[tTCL]was evaluated.The results show using thin cell layers from immature embryos cultivated in 16 light/8 dark hours achieves induction of somatic embryos.A higher percentage of embryogenic callus was obtained when tTCL segments were used as an explant source.These results produced somatic embryos from tTCL segments of an immature embryo without germinating the seed,making the process more time efficient.In addition,this technique can be used to generate somatic embryogenesis in forest species that have low germination rates.展开更多
For the mass production of Koelreuteria bipinnata var. integrifoliola with selected, hybrid or genetically engineered genotypes, one potentially desirable propagation strategy is based on embryo culture. The immature ...For the mass production of Koelreuteria bipinnata var. integrifoliola with selected, hybrid or genetically engineered genotypes, one potentially desirable propagation strategy is based on embryo culture. The immature embryo development in vitro from K. bipinnata var. integrifoliola was studied under different conditions of embryo age, basic culture media and plant growth regulators. The results show that: 1) germination rate of grade 3 embryos in immature seeds with 0.6-0.8 cm diameter was 98.9%. The germination rate of grade 2 embryos in immature seeds with 0.4-0.6 cm diameter was 77,8% and the germination rate of grade 1 embryos in immature seeds with 0.4 cm diameter was 15.6%. 2) The amounts of macroelements in MS medium had no clear effect on the germination rate of immature grade 3 embryos and had a modest effect on plantlet growth, where the best medium was MS or 1/2 MS. The rates were all greater than 90%. 3) The germination rate of grade 3 embryos was greater than 87% when the medium contained a low concentration of NAA or no plant growth regulators at all and decreased markedly when BAP alone or BAP and NAA together were added to the media. We suggest that in vitro culture of immature embryos from K. bipinnata vat. integrifoliola can be enhanced when a small amount of plant growth regulators is added. The addition of BAP has an adverse reaction to the germination and development of immature embryos.展开更多
基金Project supported by the National Natural Science Foundation of China (Grant No.11974184)。
文摘The thermal-elastic martensitic transformation from high-temperature Ni_(2)In-type hexagonal structure to low-temperature TiNiSi-type orthorhombic structure has been widely studied in MnMX(M=Ni or Co,and X=Ge or Si)alloys.However,the answer to how the orthorhombic martensite nucleates and grows within the hexagonal parent is still unclear.In this work,the hexagonal-orthorhombic martensitic transformation in a Co and Ge co-substituted MnNiSi is investigated.One can find some orthorhombic laths embedded in the hexagonal parent at a temperature above the martensitic transformation start temperature(M_(s)).With the the sample cooing to M_(s),the laths turn broader,indicating that the martensitic transformation starts from these pre-existing orthorhombic laths.Microstructure observation suggests that these pre-existing orthorhombic laths do not originate from the hexagonal-orthorhombic martensitic transformation because of the difference between atomic occupations of doping elements in the hexagonal parent and those in the preexisting orthorhombic laths.The phenomenological crystallographic theory and experimental investigations prove that the pre-existing orthorhombic lath and generated orthorhombic martensite have the same crystallography relationship to the hexagonal parent.Therefore,the orthorhombic martensite can take these pre-existing laths as embryos and grow up.This work implies that the martensitic transformation in MnNiSi_(1-x)(CoNiGe)_(x) alloy is initiated by orthorhombic embryos.
基金supported by the 948 Program of State Forestry Administration (No2009-4-26)co-sponsored by SRF for ROCS and the National Natural Science Foundation (No 30972390) of China
文摘In previous study we reported that pretreatment with plasmolysis enhanced somatic embryo formation in hypocotyls of Eleutherococcus senticosus.In the present study,the expression level of callose synthase gene in embryos of E.senticosus in response to 2,4-D,sucrose and mannitol treatments was analyzed by RT-PCR.The results show that plasmolysis pretreatment using sucrose and mannitol significantly promoted the expression of callose synthase gene.Also,the thicker cell walls of explant plasmolyzed compared with controls were observed during the somatic embryogenesis.We suggest that the callose may make the cells in epidermis separate from neighboring cells and then develop into embryogenic potential cells.
基金supported by "948" Ad-vanced Forestry Technology Introduction Program (Grant No. 2006-4-73).
文摘We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera U 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) medium supplemented with 1.0 mg·L^-1 2,4-D and 0.5 mg·L^-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg·L^-1 6-BA and 2 mg·L^-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg·L^-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dynamic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic embryos, including typical epiderma, cotyledon primordium and vascular tissue.
基金supported by the National High-tech R&D Program(863 Program)of China(2013AA102704)
文摘With embryogenic callus of Larix olgensisis, we investigated the effects of inositol, glutamine, casein hydrolysate, carbohydrate, abscisic acid and silver nitrate concentration on the maturation of the somatic embryo.Three dominant factors emerged, and we developed a response surface model based on the Box-Behnken design.We defined the optimal conditions for the maturation of somatic embryos. The contents of abscisic acid, silver nitrate, sucrose and casein hydrolysis significantly affected the amount of maturing embryos, but inositol, maltose and glutamine had no effect. By establishing a response surface model with multiple factors, we predicted that the optimal number of L. olgensis somatic embryos was 204 ± 4 gon basal medium, containing 18.28 mg Labscisic acid,5.46 mg Lsilver nitrate and 82.67 g Lsucrose. In the verification experiments, the addition of 20 mg Labscisic acid, 5 mg Lsilver nitrate and 80 g Lsucrose to BM yielded an average of 202.06 somatic embryos per gram. These results should guide large-scale breeding of L. olgensis.
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 the Natural Science Foundation of Henan Province of China(0611033300).
文摘A description of a successful direct somatic embryogenesis induction from immature zygotic embryos of a camphor tree (Cinnamomum camphora L.) is presented. After a subculture of 2-3 years, embryogenic calli could be derived from primary somatic embryos. Immature zygotic embryos were cultured on a Murashige and Skoog (MS) basal medium supplemented with a range of combinations of cytokinins (BA) and auxins (2,4-D or NAA) for somatic embryo induction. Primary somatic embryos could be induced directly in almost all PGR combinations. A positive effect of 2,4-D on somatic embryogenesis from immature zygotic embryos of camphor tree was obtained. BA at appropriate concentrations (〈 5 mg-L-1) had an effect similar to 2,4-D, whereas high concentrations (〉 5 mg·L^-1) of BA had the effect of restraining somatic embryo induction. NAA had a less positive effect on somatic embryogenesis than 2,4-D.
基金supported by grant from the Mexican Forest Service(CONAFOR)(No.11385232709—“Promotion and Operation of the Forest Genetic Resources Subsystem,within CNRGINIFAP’’)
文摘Pinus patula is a species commonly used for reforestation in Mexico.However,efficient methods for the mass production seedlings are required.Micropropagation particularly by somatic embryogenesis provides an option for the rapid multiplication of high-quality,genetically improved material.This study induces somatic embryogenesis in this species using the thin cell layer(TCL)technique.Two sources of explants(complete immature embryos;lTCL segments from immature embryos)were evaluated.The efficiency of TCL from longitudinal sections[lTCL]and transverse[tTCL]was evaluated.The results show using thin cell layers from immature embryos cultivated in 16 light/8 dark hours achieves induction of somatic embryos.A higher percentage of embryogenic callus was obtained when tTCL segments were used as an explant source.These results produced somatic embryos from tTCL segments of an immature embryo without germinating the seed,making the process more time efficient.In addition,this technique can be used to generate somatic embryogenesis in forest species that have low germination rates.
基金supported by the Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education, Beijing Forestry University, China (05–04)
文摘For the mass production of Koelreuteria bipinnata var. integrifoliola with selected, hybrid or genetically engineered genotypes, one potentially desirable propagation strategy is based on embryo culture. The immature embryo development in vitro from K. bipinnata var. integrifoliola was studied under different conditions of embryo age, basic culture media and plant growth regulators. The results show that: 1) germination rate of grade 3 embryos in immature seeds with 0.6-0.8 cm diameter was 98.9%. The germination rate of grade 2 embryos in immature seeds with 0.4-0.6 cm diameter was 77,8% and the germination rate of grade 1 embryos in immature seeds with 0.4 cm diameter was 15.6%. 2) The amounts of macroelements in MS medium had no clear effect on the germination rate of immature grade 3 embryos and had a modest effect on plantlet growth, where the best medium was MS or 1/2 MS. The rates were all greater than 90%. 3) The germination rate of grade 3 embryos was greater than 87% when the medium contained a low concentration of NAA or no plant growth regulators at all and decreased markedly when BAP alone or BAP and NAA together were added to the media. We suggest that in vitro culture of immature embryos from K. bipinnata vat. integrifoliola can be enhanced when a small amount of plant growth regulators is added. The addition of BAP has an adverse reaction to the germination and development of immature embryos.