The plants of two elfalfa （Medicago sativa L.） cultivars differing in salt tolerance were subjected to three salt treatments, 70, 140, and 210 mM NaCl for 7 days. Root, shoot, and leaf growths were inhibited by incr...The plants of two elfalfa （Medicago sativa L.） cultivars differing in salt tolerance were subjected to three salt treatments, 70, 140, and 210 mM NaCl for 7 days. Root, shoot, and leaf growths were inhibited by increased salt treatments in both cultivars, and at 140 and 210 mM salt treatments, Zhongmu 1 had significantly higher root, shoot, and leaf dry weights per plant than Deft. The malondialdehyde （MDA） accumulation in Deft was considerably greater than in Zhongmu 1, indicating a higher degree of lipid peroxidation at 140 and 210 mM salt treatments. The changes in the activity and active isoforms of antioxidant enzymes such as superoxide dismutase （SOD, EC 1.15.1.1）, catalase （CAT, EC 1.11.1.6）, peroxidase （POD, EC 1.11.1.7）, and ascorbate peroxidase （APOX, EC 1.11,1.11）, accumulation of free proline, and rate of lipid peroxidation in leaves of two alfalfa cultivars were also investigated. After stress, the activity and active isoforms of antioxidative enzymes were altered and the extent of alteration varied between the cultivar Deft and Zhongmu 1. The proline accumulation in Deft was considerably greater than in Zhongmu 1 at 210 mM salt treatment. This indicated that proline accumulation may be the result, instead of the cause, of salt tolerance.展开更多
Soil salinization and alkalization frequently co-occur in nature, but little is known about the mixed effects of salt-alkaline stresses on plants. An experiment with mixed salts （NaCI, Na2SO4, NaHCO3 and Na2CO3） and...Soil salinization and alkalization frequently co-occur in nature, but little is known about the mixed effects of salt-alkaline stresses on plants. An experiment with mixed salts （NaCI, Na2SO4, NaHCO3 and Na2CO3） and 30 salt-alkaline combinations （salinity 24-120 mmollL and pH 7.03-10.32） treating Medicago sativa seedlings was conducted. The results demonstrated that salinity and alkalinity significantly affected total biomass and biomass components of seedlings. There were interactive effects of salt composition and concentration on biomass （P 〈 0.001）. The interactions between salinity and alkalinity stresses led to changes in the root activity along the salinity gradient （P 〈 0.001）. The effects of alkalinity on seedling survival rate were more significant than those of salinity, and the seedlings demonstrated some physiological responses （leaf electrolyte leakage rate and proline content） in order to adapt to mixed salt-alkaline stresses. It was concluded that the mixed salt-alkaline stresses, which differ from either salt or alkali stress, emphasize the significant interaction between salt concentration （salinity） and salt component （alkalinity）. Further, the effects of the interaction between high alkalinity and salinity are more severe than those of either salt or alkali stress, and such a cooperative interaction results in more sensitive responses of ecological and physiological characteristics in plants.展开更多
In the Medicago genus, triterpenic saponins are bioactive secondary metabolites constitutively synthesized in the aerial and subterranean parts of plants via the isoprenoid pathway. Exploitation of saponins as pharmac...In the Medicago genus, triterpenic saponins are bioactive secondary metabolites constitutively synthesized in the aerial and subterranean parts of plants via the isoprenoid pathway. Exploitation of saponins as pharmaceutics, agrochemicals and in the food and cosmetic industries has raised interest in identifying the enzymes involved in their synthesis. We have identified a cytochrome P450 （CYP72A67） involved in hemolytic sapogenin biosynthesis by a reverse genetic TILLING approach in a Medicago truncatula ethyl- methanesulfonate （EMS） mutagenized collection. Genetic and biochemical analyses, mutant complementation, and expression of the gene in a microsome yeast system showed that CYP72A67 is responsible for hydroxylation at the C-2 position downstream of oleanolic acid synthesis. The affinity of CYP72A67 for substrates with different substitutions at multiple carbon positions was investigated in the same in vitro yeast system, and in relation to two other CYP450s （CYP72A68） responsible for the production of medicagenic acid, the main sapogenin in M. truncatula leaves and roots. Full sib mutant and wild-type plants were compared for their sapogenin profile, expression patterns of the genes involved in sapogenin synthesis, and response to inoculation with Sinorhizobium meliloti. The results obtained allowed us to revise the hemolytic sapogenin pathway in M. truncatula and contribute to highlighting the tissue specificities （leaves/roots） of sapogenin synthesis.展开更多
[Objective] The study aimed to understand the mutagenic effects of space flight on alfalfa seeds.[Method] Seeds of three lines of alfalfa were carried into orbit by the satellite 'Shijian-8' for space fight,th...[Objective] The study aimed to understand the mutagenic effects of space flight on alfalfa seeds.[Method] Seeds of three lines of alfalfa were carried into orbit by the satellite 'Shijian-8' for space fight,the indices including seed germination rate,plant height and growth rate,were measured after the seeds been retrieved.[Result] Remarkable mutagenic effects occurred on the flight seeds,which were mainly presented by abnormal cotyledon,extended variation range of plant height and growth rate.Nineteen plants with increased plant height were preliminarily screened from the T0 progenies,but whether the mutated trait could inherit should be confirmed in further study.Three flight lines of alfalfa are different in the percentages of abnormal cotyledon(10%-18%),variation range of plant height(increased 30%-150% compared to control),selected plant number with increased plant height(5-7 plants),suggesting that differences exits between the three lines in mutation efficiency.[Conclusion] Valuable mutated materials could be obtained by space flight and applied in modern agriculture.展开更多
The R2R3-MYB genes make up one of the largest transcription factor families in plants, and play regulatory roles in various biological processes such as development, metabolism and defense response. Although genome-wi...The R2R3-MYB genes make up one of the largest transcription factor families in plants, and play regulatory roles in various biological processes such as development, metabolism and defense response. Although genome-wide analyses of this gene family have been conducted in several species, R2R3-MYB genes have not been systematically analyzed in Medicago truncatula, a sequenced model legume plant. Here, we performed a comprehensive, genome-wide computational analysis of the structural characteristics, phylogeny, functions and expression patterns of M. truncatula R2R3-MYB genes. DNA binding domains are highly conserved among the 155 putative MtR2R3-MYB proteins that we identified. Chromosomal location analysis revealed that these genes were distributed across all eight chromosomes. Results showed that the expansion of the MtR2R3-MYB family was mainly attributable to segmental duplication and tandem duplication. A comprehensive classification was performed based on phylogenetic analysis of the R2R3-MYB gene families in M. truncatula, Arabidopsis thaliana and other plant species. Evolutionary relationships within clades were supported by clade-specific conserved motifs outside the MYB domain. Species-specific clades have been gained or lost during evolution, resulting in functional divergence. Also, tissue-specific expression patterns were investigated. The functions of stress response-related clades were further verified by the changes in transcript levels of representative R2R3-MYB genes upon treatment with abiotic and biotic stresses. This study is the first report on identification and characterization of R2R3-MYB gene family based on the genome of M. truncatula, and will facilitate functional analysis of this gene family in the future.展开更多
Radicle emergence and reserves mobilization are two distinct programmes that are thought to control germination. Both programs are influenced by abscissic acid （ABA） but how this hormone controls seed germination is...Radicle emergence and reserves mobilization are two distinct programmes that are thought to control germination. Both programs are influenced by abscissic acid （ABA） but how this hormone controls seed germination is still poorly known. Phenotypic and microscopic observations of the embryo axis of Medicago truncatula during germination in mitotic inhibition condition triggered by 10 μM oryzalin showed that cell division was not required to allow radicle emergence. A suppressive subtractive hybridization showed that more than 10% of up-regulated genes in the embryo axis encoded proteins related to cell-wall biosynthesis. The expression of α-expansins, pectin-esterase, xylogucan-endotransglycosidase, cellulose synthase, and extensins was monitored in the embryo axis of seeds germinated on water, constant and transitory ABA. These genes were overexpressed before completion of germination in the control and strongly inhibited by ABA. The expression was re-established in the ABA transitory-treatment after the seeds were transferred back on water and proceeded to germination. This proves these genes as contributors to the completion of germination and strengthen the idea that cell-wall loosening and remodeling in relation to cell expansion in the embryo axis is a determinant feature in germination. Our results also showed that ABA controls germination through the control of radicle emergence, namely by inhibiting cell-wall loosening and expansion.展开更多
基金supported financially by the Educational Committee of Beijing and Construction Project of Key Lab and Subject of Beijing,China (XK100190552,JD100190537)
文摘The plants of two elfalfa （Medicago sativa L.） cultivars differing in salt tolerance were subjected to three salt treatments, 70, 140, and 210 mM NaCl for 7 days. Root, shoot, and leaf growths were inhibited by increased salt treatments in both cultivars, and at 140 and 210 mM salt treatments, Zhongmu 1 had significantly higher root, shoot, and leaf dry weights per plant than Deft. The malondialdehyde （MDA） accumulation in Deft was considerably greater than in Zhongmu 1, indicating a higher degree of lipid peroxidation at 140 and 210 mM salt treatments. The changes in the activity and active isoforms of antioxidant enzymes such as superoxide dismutase （SOD, EC 1.15.1.1）, catalase （CAT, EC 1.11.1.6）, peroxidase （POD, EC 1.11.1.7）, and ascorbate peroxidase （APOX, EC 1.11,1.11）, accumulation of free proline, and rate of lipid peroxidation in leaves of two alfalfa cultivars were also investigated. After stress, the activity and active isoforms of antioxidative enzymes were altered and the extent of alteration varied between the cultivar Deft and Zhongmu 1. The proline accumulation in Deft was considerably greater than in Zhongmu 1 at 210 mM salt treatment. This indicated that proline accumulation may be the result, instead of the cause, of salt tolerance.
基金Supported by the State Key Basic Research and Development Plan(2007CB106801)the Program for Changjiang Scholars and Innovative Research Team in Universities (IRT0519)the National Natural Science Foundation of China (No. 30571318, 30600427)
文摘Soil salinization and alkalization frequently co-occur in nature, but little is known about the mixed effects of salt-alkaline stresses on plants. An experiment with mixed salts （NaCI, Na2SO4, NaHCO3 and Na2CO3） and 30 salt-alkaline combinations （salinity 24-120 mmollL and pH 7.03-10.32） treating Medicago sativa seedlings was conducted. The results demonstrated that salinity and alkalinity significantly affected total biomass and biomass components of seedlings. There were interactive effects of salt composition and concentration on biomass （P 〈 0.001）. The interactions between salinity and alkalinity stresses led to changes in the root activity along the salinity gradient （P 〈 0.001）. The effects of alkalinity on seedling survival rate were more significant than those of salinity, and the seedlings demonstrated some physiological responses （leaf electrolyte leakage rate and proline content） in order to adapt to mixed salt-alkaline stresses. It was concluded that the mixed salt-alkaline stresses, which differ from either salt or alkali stress, emphasize the significant interaction between salt concentration （salinity） and salt component （alkalinity）. Further, the effects of the interaction between high alkalinity and salinity are more severe than those of either salt or alkali stress, and such a cooperative interaction results in more sensitive responses of ecological and physiological characteristics in plants.
文摘In the Medicago genus, triterpenic saponins are bioactive secondary metabolites constitutively synthesized in the aerial and subterranean parts of plants via the isoprenoid pathway. Exploitation of saponins as pharmaceutics, agrochemicals and in the food and cosmetic industries has raised interest in identifying the enzymes involved in their synthesis. We have identified a cytochrome P450 （CYP72A67） involved in hemolytic sapogenin biosynthesis by a reverse genetic TILLING approach in a Medicago truncatula ethyl- methanesulfonate （EMS） mutagenized collection. Genetic and biochemical analyses, mutant complementation, and expression of the gene in a microsome yeast system showed that CYP72A67 is responsible for hydroxylation at the C-2 position downstream of oleanolic acid synthesis. The affinity of CYP72A67 for substrates with different substitutions at multiple carbon positions was investigated in the same in vitro yeast system, and in relation to two other CYP450s （CYP72A68） responsible for the production of medicagenic acid, the main sapogenin in M. truncatula leaves and roots. Full sib mutant and wild-type plants were compared for their sapogenin profile, expression patterns of the genes involved in sapogenin synthesis, and response to inoculation with Sinorhizobium meliloti. The results obtained allowed us to revise the hemolytic sapogenin pathway in M. truncatula and contribute to highlighting the tissue specificities （leaves/roots） of sapogenin synthesis.
基金Supported by Special Fund on Fundamental Scientific Research as Operating Expenses among Commonweal Scientific Research Institutions at Na-tional Level(Grassland Research Institute,Chinese Academy of Agricultur-al Sciences)National Key Project of Scientific and Technical Supporting Programs Funded by Ministry of Science & Technology of China(2008BADB3B04)~~
文摘[Objective] The study aimed to understand the mutagenic effects of space flight on alfalfa seeds.[Method] Seeds of three lines of alfalfa were carried into orbit by the satellite 'Shijian-8' for space fight,the indices including seed germination rate,plant height and growth rate,were measured after the seeds been retrieved.[Result] Remarkable mutagenic effects occurred on the flight seeds,which were mainly presented by abnormal cotyledon,extended variation range of plant height and growth rate.Nineteen plants with increased plant height were preliminarily screened from the T0 progenies,but whether the mutated trait could inherit should be confirmed in further study.Three flight lines of alfalfa are different in the percentages of abnormal cotyledon(10%-18%),variation range of plant height(increased 30%-150% compared to control),selected plant number with increased plant height(5-7 plants),suggesting that differences exits between the three lines in mutation efficiency.[Conclusion] Valuable mutated materials could be obtained by space flight and applied in modern agriculture.
基金supported by the National Natural Science Foundation of China(31372362)
文摘The R2R3-MYB genes make up one of the largest transcription factor families in plants, and play regulatory roles in various biological processes such as development, metabolism and defense response. Although genome-wide analyses of this gene family have been conducted in several species, R2R3-MYB genes have not been systematically analyzed in Medicago truncatula, a sequenced model legume plant. Here, we performed a comprehensive, genome-wide computational analysis of the structural characteristics, phylogeny, functions and expression patterns of M. truncatula R2R3-MYB genes. DNA binding domains are highly conserved among the 155 putative MtR2R3-MYB proteins that we identified. Chromosomal location analysis revealed that these genes were distributed across all eight chromosomes. Results showed that the expansion of the MtR2R3-MYB family was mainly attributable to segmental duplication and tandem duplication. A comprehensive classification was performed based on phylogenetic analysis of the R2R3-MYB gene families in M. truncatula, Arabidopsis thaliana and other plant species. Evolutionary relationships within clades were supported by clade-specific conserved motifs outside the MYB domain. Species-specific clades have been gained or lost during evolution, resulting in functional divergence. Also, tissue-specific expression patterns were investigated. The functions of stress response-related clades were further verified by the changes in transcript levels of representative R2R3-MYB genes upon treatment with abiotic and biotic stresses. This study is the first report on identification and characterization of R2R3-MYB gene family based on the genome of M. truncatula, and will facilitate functional analysis of this gene family in the future.
文摘Radicle emergence and reserves mobilization are two distinct programmes that are thought to control germination. Both programs are influenced by abscissic acid （ABA） but how this hormone controls seed germination is still poorly known. Phenotypic and microscopic observations of the embryo axis of Medicago truncatula during germination in mitotic inhibition condition triggered by 10 μM oryzalin showed that cell division was not required to allow radicle emergence. A suppressive subtractive hybridization showed that more than 10% of up-regulated genes in the embryo axis encoded proteins related to cell-wall biosynthesis. The expression of α-expansins, pectin-esterase, xylogucan-endotransglycosidase, cellulose synthase, and extensins was monitored in the embryo axis of seeds germinated on water, constant and transitory ABA. These genes were overexpressed before completion of germination in the control and strongly inhibited by ABA. The expression was re-established in the ABA transitory-treatment after the seeds were transferred back on water and proceeded to germination. This proves these genes as contributors to the completion of germination and strengthen the idea that cell-wall loosening and remodeling in relation to cell expansion in the embryo axis is a determinant feature in germination. Our results also showed that ABA controls germination through the control of radicle emergence, namely by inhibiting cell-wall loosening and expansion.
