Soybean （Glycine max （L.） Merr.） is a major crop that provides plant-origin protein and oil for humans and livestock. Al- though the soybean vegetative tissues and seeds provide a major source of high-quality prot...Soybean （Glycine max （L.） Merr.） is a major crop that provides plant-origin protein and oil for humans and livestock. Al- though the soybean vegetative tissues and seeds provide a major source of high-quality protein, they suffer from low con- centration of an essential sulfur-containing amino acid, methionine, which significantly limits their nutritional quality. The level of methionine is mainly controlled by the first unique enzyme of methionine synthesis, cystathione y-synthase （CGS）. Aiming to elevate methionine level in vegetative tissues and seeds, we constitutively over-expressed a feedback-insensitive Arabidopsis CGS （AtD-CGS） in soybean cultivars, Zigongdongdou （ZD） and Jilinxiaoli 1 （JX）. The levels of soluble methionine increased remarkably in leaves of transgenic soybeans compared to wild-type plants （6.6- and 7.3-fold in two transgenic ZD lines, and 3.7-fold in one transgenic JX line）. Furthermore, the total methionine contents were significantly increased in seeds of the transgenic ZD lines （1.5- to 4.8-fold increase） and the transgenic JX lines （1.3- to 2.3-fold increase） than in the wild type. The protein contents of the transgenic soybean seeds were significantly elevated compared to the wild type, suggesting that the scarcity of methionine in soybeans may limit protein accumulation in soybean seeds. The increased protein content did not alter the profile of major storage proteins in the seeds. Generally, this study provides a promising strategy to increase the levels of methionine and protein in soybean through the breeding programs.展开更多
Objective To study the relationship between polymorphism of cystathionine beta synthase （CBS） gene and development of congenital heart disease （CHD）. Methods One hundred and twenty-seven CHD case-parent triads wer...Objective To study the relationship between polymorphism of cystathionine beta synthase （CBS） gene and development of congenital heart disease （CHD）. Methods One hundred and twenty-seven CHD case-parent triads were recruited from Liaoning Province as patient group, and 129 healthy subjects without family history of birth defect were simultaneously recruited as control group together with their biological parents. For all subjects the polymorphism of CBS gene G919A locus was examined by PCR-ARMS method, Results The frequencies of three genotypes （w/w, w/m, and m/m） in control group were 27.2%, 58,4%, and 14.4%, respectively, with no significant difference in gender. A significant difference in the allele frequency was found between CHD patients and controls, the wild allele frequency was 67,9% in patients and 55.7% in controls CHD parents＇ genotype distribution was significantly different from that in controls. Further comparison of each type of CHD showed that genotype frequencies in several CHD subtypes were significantly different from those in their corresponding controls. The results of TDT analysis showed that no allele transmission disequilibrium existed in CHD nuclear families. Conclusions CBS gene G919A mutation is associated with the development of CHD, and the mutated allele may decrease the risk of CHD.展开更多
We previously showed that hydrogen sulfide(H2S)has a neuroprotective effect in the context of hypoxic ischemic brain injury in neonatal mice.However,the precise mechanism underlying the role of H2S in this situation r...We previously showed that hydrogen sulfide(H2S)has a neuroprotective effect in the context of hypoxic ischemic brain injury in neonatal mice.However,the precise mechanism underlying the role of H2S in this situation remains unclear.In this study,we used a neonatal mouse model of hypoxic ischemic brain injury and a lipopolysaccharide-stimulated BV2 cell model and found that treatment with L-cysteine,a H2S precursor,attenuated the cerebral infarction and cerebral atrophy induced by hypoxia and ischemia and increased the expression of miR-9-5p and cystathionineβsynthase(a major H2S synthetase in the brain)in the prefrontal cortex.We also found that an miR-9-5p inhibitor blocked the expression of cystathionineβsynthase in the prefrontal cortex in mice with brain injury caused by hypoxia and ischemia.Furthermore,miR-9-5p overexpression increased cystathionine-β-synthase and H2S expression in the injured prefrontal cortex of mice with hypoxic ischemic brain injury.L-cysteine decreased the expression of CXCL11,an miR-9-5p target gene,in the prefrontal cortex of the mouse model and in lipopolysaccharide-stimulated BV-2 cells and increased the levels of proinflammatory cytokines BNIP3,FSTL1,SOCS2 and SOCS5,while treatment with an miR-9-5p inhibitor reversed these changes.These findings suggest that H2S can reduce neuroinflammation in a neonatal mouse model of hypoxic ischemic brain injury through regulating the miR-9-5p/CXCL11 axis and restoringβ-synthase expression,thereby playing a role in reducing neuroinflammation in hypoxic ischemic brain injury.展开更多
The gene expression and activity of （＋）-δ-cadinene synthase during cotton development and in response to stress, as well as the spatial and temporal pattern of sesquiterpene biosynthesis, constitute one of chemica...The gene expression and activity of （＋）-δ-cadinene synthase during cotton development and in response to stress, as well as the spatial and temporal pattern of sesquiterpene biosynthesis, constitute one of chemical defense mechanisms in cotton plants. In order to explore the effects of omethoate on the cotton defense in relation to （＋）-δ-cadinene synthase and gossypol, effects of omethoate treatments on activity of （＋）-δ-cadinene synthase and gossypol content in cotton seedlings were investigated. Cotton seedlings treated with 400 mg L-1 omethoate exhibited a significant decrease in the specific activity of （＋）-δ-cadinene synthase from 12 to 120 h after treating when compared to the untreated control; significantly lower （＋）-δ-cadinene synthase activity was also observed in cotton seedlings treated with 200 mg L-1 omethoate from 72 to 120 h after treating; but for cotton seedlings treated with 100 mg L-1 omethoate, from 12 to 120 h after treating, no significant changes were observed for activity of （＋）-δ-cadinene synthase. The gossypol content in cotton seedlings treated with 100, 200 or 400 mg L-1 omethoate for different time periods showed no significant changes compared to that of the control. These results indicated that the activity of （＋）-δ-cadinene synthase in cotton seedlings in responses to exposure of omethoate at three concentrations for different time periods followed dose- and time-dependent responses to omethoate exposure. With omethoate as a chemical stress factor for cotton seedlings, the cotton defense in relation to the activity of （＋）-δ-cadinene synthase in cotton seedlings may be affected by omethoate application.展开更多
Sesquiterpenoids play an import role in the direct or indirect defense of plants.Farnesyl pyrophosphate synthases(FPSs)catalyze the biosynthesis of farnesyl pyrophosphate,which is a key precursor of farnesol and(E)-β...Sesquiterpenoids play an import role in the direct or indirect defense of plants.Farnesyl pyrophosphate synthases(FPSs)catalyze the biosynthesis of farnesyl pyrophosphate,which is a key precursor of farnesol and(E)-β-farnesene.In the current study,two FPS genes in Gossypium hirsutum,GhFPS1 and GhFPS2,were heterologously cloned and functionally characterized in a greenhouse setting.The open reading frames for full-length GhFPS1 and GhFPS2 were each 1029 nucleotides,and encoded two proteins of 342 amino acids with molecular weights of 39.4 kDa.The deduced amino acid sequences of GhFPS1–2 showed high identity to FPSs of other plants.Quantitative real-time PCR analysis revealed that GhFPS1 and GhFPS2 were highly expressed in G.hirsutum leaves,and were upregulated in methyl jasmonate(MeJA)-,methyl salicylate(MeSA)-and aphid infestation-treated cotton plants.The recombinant proteins of either GhFPS1 or GhFPS2 plus calf intestinal alkaline phosphatase could convert geranyl diphosphate(GPP)or isopentenyl diphosphate(IPP)to one major product,farnesol.Moreover,in electrophysiological response and Y-tube olfactometer assays,farnesol showed obvious attractiveness to female Aphidius gifuensis,which is an important parasitic wasp of aphids.Our findings suggest that two GhFPSs are involved in farnesol biosynthesis and they play a crucial role in indirect defense of cotton against aphid infestation.展开更多
目的热性惊厥(febrile se izure,FS)是婴幼儿时期最常见的惊厥性疾患之一,阐明其发生机制一直是该领域的重要研究课题。该课题前期的研究证明,γ-氨基丁酸B受体(-γam inobutyric ac id B receptor,GABABR)亚基和气体信号分子硫化氢(H2S...目的热性惊厥(febrile se izure,FS)是婴幼儿时期最常见的惊厥性疾患之一,阐明其发生机制一直是该领域的重要研究课题。该课题前期的研究证明,γ-氨基丁酸B受体(-γam inobutyric ac id B receptor,GABABR)亚基和气体信号分子硫化氢(H2S)均在反复热性惊厥中发挥了重要作用。该文使用GABABR激动剂ba-c lofen,抑制剂Phac lofen,探讨GABABR对FS大鼠硫化氢/胱硫醚-β-合成酶(cystath ion ine--βsynthase,CBS)体系表达的影响。方法大鼠随机分为对照组,FS组,FS+bac lofen组,FS+phac lofen组。采用热水浴诱导大鼠FS,隔日诱导1次,共10次。采用分光光度计法测定大鼠血浆中H2S含量;用原位杂交方法观察CBS mRNA表达情况;用免疫组化方法观察CBS蛋白表达情况。结果FS+bac lofen组H2S含量较FS组升高427.45±15.91μmol/L vs362.14±19.71μmol/L,同时CBS表达也较FS组增强;而FS+phac lofen组H2S含量较FS组降低189.72±21.53μmol/L vs 362.14±19.71μmol/L,同时CBS表达也较FS组减弱。结论反复热性惊厥过程中,GABABR的改变可影响H2S/CBS体系的表达。展开更多
基金supported by the Major Science and Technology Projects of China (2014ZX08004-003,2014ZX08010-004)the Natural Science Foundation of China (31471571)+2 种基金the earmarked fund for China Agriculture Research System (CARS-04)the Agricultural Science and Technology Innovation Project of CAASIsrael Science Foundation (ISF grant 231-09)
文摘Soybean （Glycine max （L.） Merr.） is a major crop that provides plant-origin protein and oil for humans and livestock. Al- though the soybean vegetative tissues and seeds provide a major source of high-quality protein, they suffer from low con- centration of an essential sulfur-containing amino acid, methionine, which significantly limits their nutritional quality. The level of methionine is mainly controlled by the first unique enzyme of methionine synthesis, cystathione y-synthase （CGS）. Aiming to elevate methionine level in vegetative tissues and seeds, we constitutively over-expressed a feedback-insensitive Arabidopsis CGS （AtD-CGS） in soybean cultivars, Zigongdongdou （ZD） and Jilinxiaoli 1 （JX）. The levels of soluble methionine increased remarkably in leaves of transgenic soybeans compared to wild-type plants （6.6- and 7.3-fold in two transgenic ZD lines, and 3.7-fold in one transgenic JX line）. Furthermore, the total methionine contents were significantly increased in seeds of the transgenic ZD lines （1.5- to 4.8-fold increase） and the transgenic JX lines （1.3- to 2.3-fold increase） than in the wild type. The protein contents of the transgenic soybean seeds were significantly elevated compared to the wild type, suggesting that the scarcity of methionine in soybeans may limit protein accumulation in soybean seeds. The increased protein content did not alter the profile of major storage proteins in the seeds. Generally, this study provides a promising strategy to increase the levels of methionine and protein in soybean through the breeding programs.
基金This work was supported by Major State Basic Research Development Program of the People's Republic of China (No. 2001CB510305).
文摘Objective To study the relationship between polymorphism of cystathionine beta synthase （CBS） gene and development of congenital heart disease （CHD）. Methods One hundred and twenty-seven CHD case-parent triads were recruited from Liaoning Province as patient group, and 129 healthy subjects without family history of birth defect were simultaneously recruited as control group together with their biological parents. For all subjects the polymorphism of CBS gene G919A locus was examined by PCR-ARMS method, Results The frequencies of three genotypes （w/w, w/m, and m/m） in control group were 27.2%, 58,4%, and 14.4%, respectively, with no significant difference in gender. A significant difference in the allele frequency was found between CHD patients and controls, the wild allele frequency was 67,9% in patients and 55.7% in controls CHD parents＇ genotype distribution was significantly different from that in controls. Further comparison of each type of CHD showed that genotype frequencies in several CHD subtypes were significantly different from those in their corresponding controls. The results of TDT analysis showed that no allele transmission disequilibrium existed in CHD nuclear families. Conclusions CBS gene G919A mutation is associated with the development of CHD, and the mutated allele may decrease the risk of CHD.
基金supported by the National Natural Science Foundation of China,Nos.82271327(to ZW),82072535(to ZW),81873768(to ZW),and 82001253(to TL).
文摘We previously showed that hydrogen sulfide(H2S)has a neuroprotective effect in the context of hypoxic ischemic brain injury in neonatal mice.However,the precise mechanism underlying the role of H2S in this situation remains unclear.In this study,we used a neonatal mouse model of hypoxic ischemic brain injury and a lipopolysaccharide-stimulated BV2 cell model and found that treatment with L-cysteine,a H2S precursor,attenuated the cerebral infarction and cerebral atrophy induced by hypoxia and ischemia and increased the expression of miR-9-5p and cystathionineβsynthase(a major H2S synthetase in the brain)in the prefrontal cortex.We also found that an miR-9-5p inhibitor blocked the expression of cystathionineβsynthase in the prefrontal cortex in mice with brain injury caused by hypoxia and ischemia.Furthermore,miR-9-5p overexpression increased cystathionine-β-synthase and H2S expression in the injured prefrontal cortex of mice with hypoxic ischemic brain injury.L-cysteine decreased the expression of CXCL11,an miR-9-5p target gene,in the prefrontal cortex of the mouse model and in lipopolysaccharide-stimulated BV-2 cells and increased the levels of proinflammatory cytokines BNIP3,FSTL1,SOCS2 and SOCS5,while treatment with an miR-9-5p inhibitor reversed these changes.These findings suggest that H2S can reduce neuroinflammation in a neonatal mouse model of hypoxic ischemic brain injury through regulating the miR-9-5p/CXCL11 axis and restoringβ-synthase expression,thereby playing a role in reducing neuroinflammation in hypoxic ischemic brain injury.
基金supported by the National Natural Science Foundation of China (30771426)the National "973"Program of China (2006CB102003)+1 种基金the Mechanisms and Detection of Insecticide Resistance of Important Pests Insects in China and Japan (NSFC and JSPS, 30911140107)the Program for China New Century Excellent Talentsin University of China (NCET-06-0113)
文摘The gene expression and activity of （＋）-δ-cadinene synthase during cotton development and in response to stress, as well as the spatial and temporal pattern of sesquiterpene biosynthesis, constitute one of chemical defense mechanisms in cotton plants. In order to explore the effects of omethoate on the cotton defense in relation to （＋）-δ-cadinene synthase and gossypol, effects of omethoate treatments on activity of （＋）-δ-cadinene synthase and gossypol content in cotton seedlings were investigated. Cotton seedlings treated with 400 mg L-1 omethoate exhibited a significant decrease in the specific activity of （＋）-δ-cadinene synthase from 12 to 120 h after treating when compared to the untreated control; significantly lower （＋）-δ-cadinene synthase activity was also observed in cotton seedlings treated with 200 mg L-1 omethoate from 72 to 120 h after treating; but for cotton seedlings treated with 100 mg L-1 omethoate, from 12 to 120 h after treating, no significant changes were observed for activity of （＋）-δ-cadinene synthase. The gossypol content in cotton seedlings treated with 100, 200 or 400 mg L-1 omethoate for different time periods showed no significant changes compared to that of the control. These results indicated that the activity of （＋）-δ-cadinene synthase in cotton seedlings in responses to exposure of omethoate at three concentrations for different time periods followed dose- and time-dependent responses to omethoate exposure. With omethoate as a chemical stress factor for cotton seedlings, the cotton defense in relation to the activity of （＋）-δ-cadinene synthase in cotton seedlings may be affected by omethoate application.
基金This work was supported by the National Natural Science Foundation of China(31772176,31672038 and 31621064)and the National Key Research and Development Program of China(2017YFDO201900 and 2017YFD0200400).
文摘Sesquiterpenoids play an import role in the direct or indirect defense of plants.Farnesyl pyrophosphate synthases(FPSs)catalyze the biosynthesis of farnesyl pyrophosphate,which is a key precursor of farnesol and(E)-β-farnesene.In the current study,two FPS genes in Gossypium hirsutum,GhFPS1 and GhFPS2,were heterologously cloned and functionally characterized in a greenhouse setting.The open reading frames for full-length GhFPS1 and GhFPS2 were each 1029 nucleotides,and encoded two proteins of 342 amino acids with molecular weights of 39.4 kDa.The deduced amino acid sequences of GhFPS1–2 showed high identity to FPSs of other plants.Quantitative real-time PCR analysis revealed that GhFPS1 and GhFPS2 were highly expressed in G.hirsutum leaves,and were upregulated in methyl jasmonate(MeJA)-,methyl salicylate(MeSA)-and aphid infestation-treated cotton plants.The recombinant proteins of either GhFPS1 or GhFPS2 plus calf intestinal alkaline phosphatase could convert geranyl diphosphate(GPP)or isopentenyl diphosphate(IPP)to one major product,farnesol.Moreover,in electrophysiological response and Y-tube olfactometer assays,farnesol showed obvious attractiveness to female Aphidius gifuensis,which is an important parasitic wasp of aphids.Our findings suggest that two GhFPSs are involved in farnesol biosynthesis and they play a crucial role in indirect defense of cotton against aphid infestation.
文摘目的热性惊厥(febrile se izure,FS)是婴幼儿时期最常见的惊厥性疾患之一,阐明其发生机制一直是该领域的重要研究课题。该课题前期的研究证明,γ-氨基丁酸B受体(-γam inobutyric ac id B receptor,GABABR)亚基和气体信号分子硫化氢(H2S)均在反复热性惊厥中发挥了重要作用。该文使用GABABR激动剂ba-c lofen,抑制剂Phac lofen,探讨GABABR对FS大鼠硫化氢/胱硫醚-β-合成酶(cystath ion ine--βsynthase,CBS)体系表达的影响。方法大鼠随机分为对照组,FS组,FS+bac lofen组,FS+phac lofen组。采用热水浴诱导大鼠FS,隔日诱导1次,共10次。采用分光光度计法测定大鼠血浆中H2S含量;用原位杂交方法观察CBS mRNA表达情况;用免疫组化方法观察CBS蛋白表达情况。结果FS+bac lofen组H2S含量较FS组升高427.45±15.91μmol/L vs362.14±19.71μmol/L,同时CBS表达也较FS组增强;而FS+phac lofen组H2S含量较FS组降低189.72±21.53μmol/L vs 362.14±19.71μmol/L,同时CBS表达也较FS组减弱。结论反复热性惊厥过程中,GABABR的改变可影响H2S/CBS体系的表达。
