Objective:To investigate the effect of saponin from the seed of Litchi chinensis Sonn(SLS)on the growth and apoptosis of human kidney epithelial cells(HKC)cultured in high glucose.Methods:HKC were cultured in DMEM/F12...Objective:To investigate the effect of saponin from the seed of Litchi chinensis Sonn(SLS)on the growth and apoptosis of human kidney epithelial cells(HKC)cultured in high glucose.Methods:HKC were cultured in DMEM/F12 medium supplemented with 30 mmol/L glucose and treated with or without SLS.In the normal group,isometric DMEM/F12 medium with 5.5mmol/L glucose was added.The secretion of TGF-β1 and fibronectin(FN)were detected by ELISA.Cell apoptosis was detected by the method of Annexin V-FITC/PI double staining.Western blot was used to detect the level of suppressor of cytokine signaling-1(SOCS-1).Results:The result of ELISA showed that the secretion of TGF-β1 and FN was decreased in SLS groups compared with those in 30 mmol/L glucose treated group(P<0.05).There were more cells apoptosis in 30 mmol/L glucose treated group than that in the normal group(P<0.01).Compared with the 30 mmol/L glucose treated group,the apoptosis of HKC were significantly decreased in SLS groups(P<0.01).Western blot showed that the level of SOCS-1 in high glucose+SLS group was decreased(P<0.01),compared with the high glucose group.Conclusion:SLS can reduce the secretion of TGF-β1 and FN in HKC by reducing the deposition of extracellular matrix.SLS also significantly reduced the apoptosis of HKC by inhibiting the level of SOCS-1.These results suggest the roles of SLS in preventing the progress of glomerular sclerosis.展开更多
Polyamines play an important role in plant response to abiotic stress. S-adenosyl-1-methionine decarboxylase (SAMDC) is one of the key regulatory enzymes in the biosynthesis of polyamines. In order to better understan...Polyamines play an important role in plant response to abiotic stress. S-adenosyl-1-methionine decarboxylase (SAMDC) is one of the key regulatory enzymes in the biosynthesis of polyamines. In order to better understand the effect of regulation of polyamine biosynthesis on the shelf life improvement of litchi fruit, SAMDC cDNA isolated from Datura stramonium cloned in pBI121 was introduced into litchi genome by means of Agrobacterium tumefaciens through zygote disc transformation. Transgene and its expression are confirmed by Southern and Northern blot analyses, respectively. Transgenic plants expressing Datura SAMDC produced 1.7- to 2.4-fold higher levels of spermidine and spermine than wildtype plants under normal environmental condition, which indicated that the transgenic litchi presented an enhanced polyamines synthesis compared to wildtype plants. Our results demonstrated clearly that increasing polyamine biosynthesis in plants may be a means of creating improved fruit shelf life germplasm.展开更多
Litchi(Litchi chinensis Sonn.)is a type of commercially prevalent subtropical and tropical fruit.Since litchi has a highly heterozygous genetic background and a long reproductive cycle,conventional breeding methods(su...Litchi(Litchi chinensis Sonn.)is a type of commercially prevalent subtropical and tropical fruit.Since litchi has a highly heterozygous genetic background and a long reproductive cycle,conventional breeding methods(such as hybridization)have limited ability to nurture new litchi cultivars.Here,an efficient and stable Agrobacterium tumefaciens-mediated genetic transformation of embryogenic callus was established in‘Feizixiao’litchi.Transgenic materials were verified using polymerase chain reaction(PCR)analysis,β-glucuronidase(GUS)assay,and green fluorescent protein(GFP)assay.To implement the technology of the Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)/associated protein 9(CRISPR/Cas9)technology in‘Feizixiao’litchi and verify the validity of these transformation systems,the litchi polyphenol oxidase gene(LcPPO,JF926153)was knocked out.Various categories of mutations,covering base insertions,deletions,and substitutions,were found in transgenic materials via sequence analysis.The transformation system achieved high feasibility and efficiency,and the system of CRISPR/Cas9 was successfully employed to edit genes in‘Feizixiao’litchi.This work provides an essential foundation for investigating the functions of genes and accelerating litchi genetic improvement.展开更多
Using PCR degenerate primers, designed with reference to the sequences of the conserved amino acids of known expansins, to amplify cDNA fragments in litchi fruit by RT-PCR, two different cDNA fragments , named as Lc-E...Using PCR degenerate primers, designed with reference to the sequences of the conserved amino acids of known expansins, to amplify cDNA fragments in litchi fruit by RT-PCR, two different cDNA fragments , named as Lc-Exp1 and Lc-Exp2 , were cloned. Lc-Exp1 and Lc-Exp2 was respectively composed of 531 bp encoding 177 amino acids and 537 bp encoding 179 amino acids. Eight cysteine residues and three tryp-tophan residues, which is supposed to be the characteristics of expansins, are conserved in both Lc-Exp1 and c-Exp2. In addition, the homology between the two expansins is 71. 6% at nucleotide acid sequences and 76.3% at amino acid sequences. The homology of Lc-Exp1 with Fa-Exp2 or Pp-Exp1 was 92.7% or 92.1%, but that of Lc-Exp2 with Fa-Exp2 or Pp-Exp1 was only 77. 4% or 76.3% at amino acid sequences.展开更多
[Objective] Pericarp browning in the postharvest litchi significantly reduced its commercial value and limited the expanding of litchi markets. Physiological changes during the process of pericarp browning were determ...[Objective] Pericarp browning in the postharvest litchi significantly reduced its commercial value and limited the expanding of litchi markets. Physiological changes during the process of pericarp browning were determined in order to identify the underlying mechanisms. [Method] Matured Feizixiao fruits were stored at 25 ℃ and 70%±5% relative humidity. The physiological changes happened in pericarp during storage were tested at an 8-hour interval. [Result] The fruit of Feizixiao (Litchi chinensis Sonn. cv Feizixiao) turned completely brown within 72 h after being harvested under the experimental conditions. Sharp increase of the browning index occurred from 48 to 64 hours after harvest (HAH). With the browning of pericarp,water content of the whole fruit and pericarp decreased continuingly. In contrast,there were no significant changes in the water content of pulp during the same period. MDA content,pH value and relative leakage rate of pericarp were increased during storage. Most of pigment contents including anthocyanin,flavonoid,phenols,chlorophyll a and total chlorophyll decreased. POD activity was initially increased in 32 HAH and then decreased afterwards. PPO activity was decreased continuously,while the activities of catalase and superoxide dismutase exhibited the pattern of 'increasing-decreasing-increasing' as the storage time progressed. Correlation,stepwise regression and path analyses showed that water loss of pericarp was the major factor of pericarp browning. Principal and cluster analyses showed that there were two stages of pericarp browning during the course of litchi storage. [Conclusion] Water status of pericarp was the most important factor affecting pericarp browning. The pericarp browning happened by stages,which was mainly determined by the water loss of pericarp.展开更多
The regulatory role of calcium in fertility of pollen and pistil under simulated acid rain was investigated. The germination percentage of pollen treated with acid rain of pH 4.5 was 9.42% lower than that of control, ...The regulatory role of calcium in fertility of pollen and pistil under simulated acid rain was investigated. The germination percentage of pollen treated with acid rain of pH 4.5 was 9.42% lower than that of control, and that of pH 3.5, pH 2.5 and pH 1.5 were 22.47%, 45.49% and 71.62%, respectively. Simultaneously, the injury character of pollen was obviously observed when flowers were treated with acid rain of pH 3.5. The difference in fruit setting rate between the female flower treated with acid rain of pH 4.0 and the control was significant at p 〈 0.05. Ca(NO3 )5 of 0.2-0.4 mmol/L could promote pollen germination under the stress of acid rain. The beneficial function was reduced when calcium concentration surpassed 0.8 mmol/L. Spraying 2 mmol/L Ca(NO3 )5 reduced the injury of acid rain to pistil and increased fruit-setting rate significantly. The physiological importance of calcium during pollen germination and pistil development was also discussed.展开更多
建立不同品种荔枝核的定性分析方法,并探讨不同品种荔枝核间共有成分的变化规律。采用超高效液相色谱-串联四极杆-飞行时间质谱技术(ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry,UPLC...建立不同品种荔枝核的定性分析方法,并探讨不同品种荔枝核间共有成分的变化规律。采用超高效液相色谱-串联四极杆-飞行时间质谱技术(ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry,UPLC-Q-TOF/MS)[ACQUITY UPLC HSS T3色谱柱;流动相以0.1%甲酸水溶液为A,以0.1%甲酸乙腈溶液为B,进行梯度洗脱;体积流量0.4 mL/min;柱温30℃,电喷雾离子源负离子模式(ESI-)]分析不同品种荔枝核中共有化学成分及含量高低。结果显示4种不同品种的荔枝核供试品共检识化合物99个,其中共有成分有9个,分别是维生素C、喇叭茶醇、去氢白菖蒲烯、别香橙烯、芍药二酮、缬氨酸、紫苏醇、环-(亮氨酸-异亮氨酸)、甲基麦冬黄烷酮B。展开更多
单锌指DNA结合蛋白(DNA binding with one finger,Dof)是植物中特有的一类转录因子,在植物生长发育与非生物胁迫响应中发挥非常重要的作用。利用妃子笑荔枝果实发育RNA-seq数据库,采用生物信息学分析方法,对荔枝Dof(LcDof)基因家族的基...单锌指DNA结合蛋白(DNA binding with one finger,Dof)是植物中特有的一类转录因子,在植物生长发育与非生物胁迫响应中发挥非常重要的作用。利用妃子笑荔枝果实发育RNA-seq数据库,采用生物信息学分析方法,对荔枝Dof(LcDof)基因家族的基本理化特性,亚细胞定位,蛋白质保守结构域,进化关系等进行分析,同时对LcDof基因家族在果实发育中的表达情况进行分析。结果表明:荔枝中包含有19个LcDof基因家族成员,LcDof编码蛋白范围在157~497个氨基酸残基(Amino acid,aa),对应的分子量为17.70~54.35 kDa,等电点(pI)范围为4.49~9.42;预测LcDof家族成员亚细胞定位均定位于细胞核。系统进化关系分析表明,LcDof基因家族分为4组(GroupⅠ~GroupⅣ),LcDof在不同发育阶段的表达模式不同,其中LcDof7、LcDof9、LcDof12、LcDof15在果肉不同发育时期表达量较高,而LcDof3、LcDof10、LcDof16、LcDof17、LcDof19表达量较低。展开更多
With the bioactivity-guided method,a new flavanone glycoside,together with nine known flavonoids were isolated from 50% aqueous ethanol of litchi(Litchi chinensis Sonn.) seeds.The chemical structure of the new compo...With the bioactivity-guided method,a new flavanone glycoside,together with nine known flavonoids were isolated from 50% aqueous ethanol of litchi(Litchi chinensis Sonn.) seeds.The chemical structure of the new compound was elucidated via 1D and 2D nuclear magnetic resonance(NMR) techniques and mass spectrometry to be (2S)-pinocembrin-7-O-(6"-O-α-L-arabinosyl-β-D-glucopyranoside)(1),and the nine known compounds were determined to be quercetin(2),phlorhizin(3),pinocembrin-7-O-glucoside(4),kaempferol-7-O-β-D-glucopyranoside(5),onychin(6),nairutin(7),narcissin(8),pinocembrin-7-O-[(6"-O-β-D-glucopyranoside)-β-D-glucopyranoside](9) and pinocembrin-7-O-[(2",6"-di-O-α-L-rhamnopyranosyl)-β-D-glucopyranoside](10).Some of the isolated flavonoids were tested for their inhibitory effects on α-glucosidase.And compounds 2 and 3 showed stronger inhibitory activity than positive control.展开更多
基金National Natural Science Foundation of China(No.81374025)the Education Department of Jilin Province“13th Five-Year”science and technology research project.
文摘Objective:To investigate the effect of saponin from the seed of Litchi chinensis Sonn(SLS)on the growth and apoptosis of human kidney epithelial cells(HKC)cultured in high glucose.Methods:HKC were cultured in DMEM/F12 medium supplemented with 30 mmol/L glucose and treated with or without SLS.In the normal group,isometric DMEM/F12 medium with 5.5mmol/L glucose was added.The secretion of TGF-β1 and fibronectin(FN)were detected by ELISA.Cell apoptosis was detected by the method of Annexin V-FITC/PI double staining.Western blot was used to detect the level of suppressor of cytokine signaling-1(SOCS-1).Results:The result of ELISA showed that the secretion of TGF-β1 and FN was decreased in SLS groups compared with those in 30 mmol/L glucose treated group(P<0.05).There were more cells apoptosis in 30 mmol/L glucose treated group than that in the normal group(P<0.01).Compared with the 30 mmol/L glucose treated group,the apoptosis of HKC were significantly decreased in SLS groups(P<0.01).Western blot showed that the level of SOCS-1 in high glucose+SLS group was decreased(P<0.01),compared with the high glucose group.Conclusion:SLS can reduce the secretion of TGF-β1 and FN in HKC by reducing the deposition of extracellular matrix.SLS also significantly reduced the apoptosis of HKC by inhibiting the level of SOCS-1.These results suggest the roles of SLS in preventing the progress of glomerular sclerosis.
文摘Polyamines play an important role in plant response to abiotic stress. S-adenosyl-1-methionine decarboxylase (SAMDC) is one of the key regulatory enzymes in the biosynthesis of polyamines. In order to better understand the effect of regulation of polyamine biosynthesis on the shelf life improvement of litchi fruit, SAMDC cDNA isolated from Datura stramonium cloned in pBI121 was introduced into litchi genome by means of Agrobacterium tumefaciens through zygote disc transformation. Transgene and its expression are confirmed by Southern and Northern blot analyses, respectively. Transgenic plants expressing Datura SAMDC produced 1.7- to 2.4-fold higher levels of spermidine and spermine than wildtype plants under normal environmental condition, which indicated that the transgenic litchi presented an enhanced polyamines synthesis compared to wildtype plants. Our results demonstrated clearly that increasing polyamine biosynthesis in plants may be a means of creating improved fruit shelf life germplasm.
基金supported by grants from the National Key R&D Program of China(Grant No.2019YFD1000900)the Hainan Province Science and Technology Special Fund(Grant No.ZDYF2022XDNY253)the earmarked fund for CARS(Grant No.CARS-32-01)。
文摘Litchi(Litchi chinensis Sonn.)is a type of commercially prevalent subtropical and tropical fruit.Since litchi has a highly heterozygous genetic background and a long reproductive cycle,conventional breeding methods(such as hybridization)have limited ability to nurture new litchi cultivars.Here,an efficient and stable Agrobacterium tumefaciens-mediated genetic transformation of embryogenic callus was established in‘Feizixiao’litchi.Transgenic materials were verified using polymerase chain reaction(PCR)analysis,β-glucuronidase(GUS)assay,and green fluorescent protein(GFP)assay.To implement the technology of the Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)/associated protein 9(CRISPR/Cas9)technology in‘Feizixiao’litchi and verify the validity of these transformation systems,the litchi polyphenol oxidase gene(LcPPO,JF926153)was knocked out.Various categories of mutations,covering base insertions,deletions,and substitutions,were found in transgenic materials via sequence analysis.The transformation system achieved high feasibility and efficiency,and the system of CRISPR/Cas9 was successfully employed to edit genes in‘Feizixiao’litchi.This work provides an essential foundation for investigating the functions of genes and accelerating litchi genetic improvement.
文摘Using PCR degenerate primers, designed with reference to the sequences of the conserved amino acids of known expansins, to amplify cDNA fragments in litchi fruit by RT-PCR, two different cDNA fragments , named as Lc-Exp1 and Lc-Exp2 , were cloned. Lc-Exp1 and Lc-Exp2 was respectively composed of 531 bp encoding 177 amino acids and 537 bp encoding 179 amino acids. Eight cysteine residues and three tryp-tophan residues, which is supposed to be the characteristics of expansins, are conserved in both Lc-Exp1 and c-Exp2. In addition, the homology between the two expansins is 71. 6% at nucleotide acid sequences and 76.3% at amino acid sequences. The homology of Lc-Exp1 with Fa-Exp2 or Pp-Exp1 was 92.7% or 92.1%, but that of Lc-Exp2 with Fa-Exp2 or Pp-Exp1 was only 77. 4% or 76.3% at amino acid sequences.
基金Supported by National Science Foundation of China ( GrantNo.30460085, 30960233)Open Foundation of Provincial Key Laboratory for Fruit and Vegetable Preservation of Hainan ( GrantNo. CH001)National Non-profit Institute Grant (ITBBZD2007-3-1)~~
文摘[Objective] Pericarp browning in the postharvest litchi significantly reduced its commercial value and limited the expanding of litchi markets. Physiological changes during the process of pericarp browning were determined in order to identify the underlying mechanisms. [Method] Matured Feizixiao fruits were stored at 25 ℃ and 70%±5% relative humidity. The physiological changes happened in pericarp during storage were tested at an 8-hour interval. [Result] The fruit of Feizixiao (Litchi chinensis Sonn. cv Feizixiao) turned completely brown within 72 h after being harvested under the experimental conditions. Sharp increase of the browning index occurred from 48 to 64 hours after harvest (HAH). With the browning of pericarp,water content of the whole fruit and pericarp decreased continuingly. In contrast,there were no significant changes in the water content of pulp during the same period. MDA content,pH value and relative leakage rate of pericarp were increased during storage. Most of pigment contents including anthocyanin,flavonoid,phenols,chlorophyll a and total chlorophyll decreased. POD activity was initially increased in 32 HAH and then decreased afterwards. PPO activity was decreased continuously,while the activities of catalase and superoxide dismutase exhibited the pattern of 'increasing-decreasing-increasing' as the storage time progressed. Correlation,stepwise regression and path analyses showed that water loss of pericarp was the major factor of pericarp browning. Principal and cluster analyses showed that there were two stages of pericarp browning during the course of litchi storage. [Conclusion] Water status of pericarp was the most important factor affecting pericarp browning. The pericarp browning happened by stages,which was mainly determined by the water loss of pericarp.
文摘The regulatory role of calcium in fertility of pollen and pistil under simulated acid rain was investigated. The germination percentage of pollen treated with acid rain of pH 4.5 was 9.42% lower than that of control, and that of pH 3.5, pH 2.5 and pH 1.5 were 22.47%, 45.49% and 71.62%, respectively. Simultaneously, the injury character of pollen was obviously observed when flowers were treated with acid rain of pH 3.5. The difference in fruit setting rate between the female flower treated with acid rain of pH 4.0 and the control was significant at p 〈 0.05. Ca(NO3 )5 of 0.2-0.4 mmol/L could promote pollen germination under the stress of acid rain. The beneficial function was reduced when calcium concentration surpassed 0.8 mmol/L. Spraying 2 mmol/L Ca(NO3 )5 reduced the injury of acid rain to pistil and increased fruit-setting rate significantly. The physiological importance of calcium during pollen germination and pistil development was also discussed.
文摘建立不同品种荔枝核的定性分析方法,并探讨不同品种荔枝核间共有成分的变化规律。采用超高效液相色谱-串联四极杆-飞行时间质谱技术(ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry,UPLC-Q-TOF/MS)[ACQUITY UPLC HSS T3色谱柱;流动相以0.1%甲酸水溶液为A,以0.1%甲酸乙腈溶液为B,进行梯度洗脱;体积流量0.4 mL/min;柱温30℃,电喷雾离子源负离子模式(ESI-)]分析不同品种荔枝核中共有化学成分及含量高低。结果显示4种不同品种的荔枝核供试品共检识化合物99个,其中共有成分有9个,分别是维生素C、喇叭茶醇、去氢白菖蒲烯、别香橙烯、芍药二酮、缬氨酸、紫苏醇、环-(亮氨酸-异亮氨酸)、甲基麦冬黄烷酮B。
文摘单锌指DNA结合蛋白(DNA binding with one finger,Dof)是植物中特有的一类转录因子,在植物生长发育与非生物胁迫响应中发挥非常重要的作用。利用妃子笑荔枝果实发育RNA-seq数据库,采用生物信息学分析方法,对荔枝Dof(LcDof)基因家族的基本理化特性,亚细胞定位,蛋白质保守结构域,进化关系等进行分析,同时对LcDof基因家族在果实发育中的表达情况进行分析。结果表明:荔枝中包含有19个LcDof基因家族成员,LcDof编码蛋白范围在157~497个氨基酸残基(Amino acid,aa),对应的分子量为17.70~54.35 kDa,等电点(pI)范围为4.49~9.42;预测LcDof家族成员亚细胞定位均定位于细胞核。系统进化关系分析表明,LcDof基因家族分为4组(GroupⅠ~GroupⅣ),LcDof在不同发育阶段的表达模式不同,其中LcDof7、LcDof9、LcDof12、LcDof15在果肉不同发育时期表达量较高,而LcDof3、LcDof10、LcDof16、LcDof17、LcDof19表达量较低。
基金Supported by the National Natural Science Foundation of China(No.30873370) and the Doctor Fund of Jilin Agricultural University of China(No.201210).
文摘With the bioactivity-guided method,a new flavanone glycoside,together with nine known flavonoids were isolated from 50% aqueous ethanol of litchi(Litchi chinensis Sonn.) seeds.The chemical structure of the new compound was elucidated via 1D and 2D nuclear magnetic resonance(NMR) techniques and mass spectrometry to be (2S)-pinocembrin-7-O-(6"-O-α-L-arabinosyl-β-D-glucopyranoside)(1),and the nine known compounds were determined to be quercetin(2),phlorhizin(3),pinocembrin-7-O-glucoside(4),kaempferol-7-O-β-D-glucopyranoside(5),onychin(6),nairutin(7),narcissin(8),pinocembrin-7-O-[(6"-O-β-D-glucopyranoside)-β-D-glucopyranoside](9) and pinocembrin-7-O-[(2",6"-di-O-α-L-rhamnopyranosyl)-β-D-glucopyranoside](10).Some of the isolated flavonoids were tested for their inhibitory effects on α-glucosidase.And compounds 2 and 3 showed stronger inhibitory activity than positive control.