Soil salinity badly affects agriculture productivity through accumulation of salts in upper layers of soils. The harmful effects of salts in arable lands have influenced modern as well as ancient civilizations. A pot ...Soil salinity badly affects agriculture productivity through accumulation of salts in upper layers of soils. The harmful effects of salts in arable lands have influenced modern as well as ancient civilizations. A pot study was carried out to test the performance of two PGPR isolates (KS 8, KS 28) on sunflower (SMH-0917) under different salinity levels (8, 10 and 12 dS·m-1). These salinity levels were developed by adding calculated amount of salts (NaCl, Na2SO4, CaCl2 and MgSO4) with ratio of 3:4:2:1. The bacterial strains KS 8 and KS 28 were applied separately in two treatments while third treatment was co-inoculation (KS mix). Completely randomized experimental design (CRD) was used and data were collected at flowering stage about pre-decided plant growth parameters (plant height, shoot dry weight and root dry weight). The bacterial isolate KS 8 showed an increase of 26, 102% and 83% in plant height, shoot dry weight and root dry weight at EC 8 dS·m-1, while this improvement was 67%, 163% and 296% at EC 10 dS·m-1, however an increase of 100%, 74% and 382% was recorded over control respectively at EC 12 dS·m-1. Similarly isolate KS 28 exhibited an increase of 14%, 69% and 54% in plant height;shoot dry weight and root dry weight at EC 8 dS·m-1, whereas this improvement was 56%, 163% and 188% at EC 10 dS·m-1, while an increase of 60%, 41% and 282% was registered respectively over control at EC 12 dS·m-1. The increase due to mixture treatments was 4%, 41% and 16% in plant height, shoot dry weight and root dry weight at EC 8 dS·m-1, while an increase of 33%, 57% and 100% at EC 10 dS·m-1, whereas an improvement of 53%, 33% and 164% respectively was noted at EC 12 dS·m-1 over un-inoculated. The isolate KS 8 performed better than KS 28 and mixture treatment. These two PGPR strains could be used to mitigate the adverse impact caused by salinity stress on sunflower.展开更多
To elucidate the relationship between ethylene evolution from the grains and the appearance quality of rice, ten different rice genotypes were used to determine the ethylene evolution rate, 1-aminocylopropane-1-carbox...To elucidate the relationship between ethylene evolution from the grains and the appearance quality of rice, ten different rice genotypes were used to determine the ethylene evolution rate, 1-aminocylopropane-1-carboxylic acid （ACC） concentration in grains during grain filling and the appearance quality of rice, and the effects of chemical regulators on concentrations of ethylene and ACC in the grains during grain filling were also investigated to verify the roles of ethylene in the rice quality formation. The ethylene evolution rates and ACC concentrations in grains during the mid and late grain filling stages were very significantly and positively correlated with chalky kernel percentage and chalkiness. The cultivars with a low ACC concentration in grains exhibited a close amyloplast arrangement and small space between starch granules, whereas those with a high ACC concentration in grains showed a loose arrangement and wide space between the granules. Application of 1 μmol/L ACC to panicles at mid and late grain filling stages significantly loosened amyloplast arrangement and increased chalky kernel percentage, chalky area and chalkiness, and the results were reversed when 1 μmol/L amino-ethoxyvinylglycine, an inhibitor of ACC synthesis enzyme, was applied to panicles. A practice of moderate dry-wet alternate irrigation reduced ethylene evolution and ACC concentration in grains and thereby reduced chalkiness. The results suggested that ethylene and ACC in grains play an important role in the endosperm structure and appearance quality of rice, and the appearance quality would be improved by reducing ethylene evolution and ACC in grains through either variety breeding and selection, or chemical regulations or cultivation techniques.展开更多
1-H-Pyrrole-2-carboxylic acid [2-（naphthalen-1-ylamino）-ethyl]-amide has been synthesized and characterized. Its crystal is of monoclinic, space group P2 1/n with a = 5.930（6）, b = 12.144（13）, c = 20.10（2） , ...1-H-Pyrrole-2-carboxylic acid [2-（naphthalen-1-ylamino）-ethyl]-amide has been synthesized and characterized. Its crystal is of monoclinic, space group P2 1/n with a = 5.930（6）, b = 12.144（13）, c = 20.10（2） , A, β = 95.709（17）°, V= 1441（3） ,A, Z= 4, C17H17N3O, Mr= 279.34, Dc= 1.288 g/cm^3, F（000） = 592, μ（MoKa） = 0.083 mm^-1, S = 1.019, R = 0.0473 and wR = 0.1181 for 1713 observed reflections with I 〉 2 σ（I）. X-ray diffraction reveals that two molecules of the title compound form a dimer through a pair of N-H…O hydrogen bonds.展开更多
BACKGROUND 3,6-dichlorobenzo[b]thiophene-2-carboxylic acid(BT2)is a benzothiophene carboxylate derivative that can suppress the catabolism of branched-chain amino acid(BCAA)-associated mammalian target of rapamycin co...BACKGROUND 3,6-dichlorobenzo[b]thiophene-2-carboxylic acid(BT2)is a benzothiophene carboxylate derivative that can suppress the catabolism of branched-chain amino acid(BCAA)-associated mammalian target of rapamycin complex 1(mTORC1)activation.Previous studies have demonstrated the therapeutic effects of BT2 on arthritis,liver cancer,and kidney injury.However,the effects of BT2 on ulcerative colitis(UC)are unknown.AIM To investigate the anti-UC effects of BT2 and the underlying mechanism.METHODS Mouse UC models were created through the administration of 3.5%dextran sodium sulfate(DSS)for 7 d.The mice in the treated groups were administered salazosulfapyridine(300 mg/kg)or BT2(20 mg/kg)orally from day 1 to day 7.At the end of the study,all of the mice were sacrificed,and colon tissues were removed for hematoxylin and eosin staining,immunoblot analyses,and immunohistochemical assays.Cytokine levels were measured by flow cytometry.The contents of BCAAs including valine,leucine,and isoleucine,in mouse serum were detected by liquid chromatography-tandem mass spectrometry,and the abundance of intestinal flora was analyzed by 16S ribosomal DNA sequencing.RESULTS Our results revealed that BT2 significantly ameliorated the inflammatory symptoms and pathological damage induced by DSS in mice.BT2 also reduced the production of the proinflammatory cytokines interleukin 6(IL-6),IL-9,and IL-2 and increased the anti-inflammatory cytokine IL-10 level.In addition,BT2 notably improved BCAA catabolism and suppressed mTORC1 activation and cyclooxygenase-2 expression in the colon tissues of UC mice.Furthermore,highthroughput sequencing revealed that BT2 restored the gut microbial abundance and diversity in mice with colitis.Compared with the DSS group,BT2 treatment increased the ratio of Firmicutes to Bacteroidetes and decreased the abundance of Enterobacteriaceae and Escherichia-Shigella.CONCLUSION Our results indicated that BT2 significantly ameliorated DSS-induced UC and that the latent mechanism involved the suppression of BCAA-associated mTORC1 activation and modulation of the intestinal flora.展开更多
1-Aminocyclopropane-1-carboxylic acid （ACC） synthase （ACS） is the principal enzyme in phytohormone ethylene biosynthesis. Previous studies have shown that the hypervariable C-terminus of ACS is proteolytically pro...1-Aminocyclopropane-1-carboxylic acid （ACC） synthase （ACS） is the principal enzyme in phytohormone ethylene biosynthesis. Previous studies have shown that the hypervariable C-terminus of ACS is proteolytically processed in vivo. However, the protease responsible for this has not yet been identified. In the present study, we investigated the processing of the 55-kDa full-length tomato ACS （LeACS2） into 52-, 50- and 49-kDa truncated isoforms in ripening tomato （Lycopersicon esculentum Mill. cv. Cooperation 903） fruit using the sodium dodecyl sulfate-boiling method. Meanwhile, an LeACS2-processing protease was purified via multi-step column chromatography from tomato fruit. Subsequent biochemical analysis of the 64-kDa purified protease revealed that it is a metalloprotease active at multiple cleavage sites within the hypervariable C-terminus of LeACS2. N-terminal sequencing and matrix-assisted laser desorption/ionization time-of-flight analysis indicated that the LeACS2-processing metalloprotease cleaves at the C-terminal sites Lys^438, Glu^447, Lys^448, Asn^456, Ser^460, Ser^462, Lys^463, and Leu^474, but does not cleave the N- terminus of LeACS2. Four C-terminus-deleted （26-50 amino acids） LeACS2 fusion proteins were overproduced and subjected to proteolysis by this metalloprotease to identify the multiple cleavage sites located on the N-terminal side of the phosphorylation site Ser^460. The results indisputably confirmed the presence of cleavage sites within the region between the α-helix domain （H14） and Ser^460 for this metalloprotease. Furthermore, the resulting C-terminally truncated LeACS2 isoforms were active enzymatically. Because this protease could produce LeACS2 isoforms in vitro similar to those detected in vivo, it is proposed that this metalloprotease may be involved in the proteolysis of LeACS2 in vivo.展开更多
Summary Although it has been reported previously that ethylene plays a critical role in sex determination in cucurbit species, how the andromonoecy that carries both the male and hermaphroditic flowers is determined i...Summary Although it has been reported previously that ethylene plays a critical role in sex determination in cucurbit species, how the andromonoecy that carries both the male and hermaphroditic flowers is determined in watermelon is still unknown. Here we showed that the watermelon gene 1-aminocyclopropane-1-carboxylate syn- thase 4 CCitACS4）, expressed specifically in carpel primor- dia, determines the andromonoecy in watermelon. Among four single nucleotide polymorphism （SNPs） and one lnDel identified in the coding region of CitACS4, the C364W mutation located in the conserved box 6 was co- segregated with andromonoecy. Enzymatic analyses showed that the C364W mutation caused a reduced activity in CitACS4. We believe that the reduced CitACS4 activity may hamper the programmed cell death in stamen primordia, leading to the formation of hermaphroditic flowers.展开更多
目的探讨健脾疏肝丸对非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)大鼠肝X受体(liver X receptorα,LXRα)-固醇调节元件结合蛋白-1(sterolregulatory element-binding protein-1,SREBP-1)-脂肪酸合成酶(fatty acid s...目的探讨健脾疏肝丸对非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)大鼠肝X受体(liver X receptorα,LXRα)-固醇调节元件结合蛋白-1(sterolregulatory element-binding protein-1,SREBP-1)-脂肪酸合成酶(fatty acid synthase,FAS)信号转导通路的影响。方法将32只无特定病原体(specific pathogen free,SPF)SD雄性大鼠按照随机数字表法分为正常组、模型组、健脾疏肝丸组和易善复组,每组8只。正常组进食普通饲料,其他组进食高脂饲料,健脾疏肝丸组给予4.86 g/(kg·d)灌胃,易善复组给予0.123 g/(kg·d)灌胃,正常组和模型组给予等量蒸馏水灌胃。灌胃与造模同时进行,共8周。检测大鼠血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、高密度脂蛋白(high-density lipoprotein cholesterol,HDL-C)、低密度脂蛋白(low-density lipoprotein cholesterol,LDL-C)、甘油三酯(triglycerides,TG)、总胆固醇(total cholesterol,TC)、肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)及白细胞介素-6(interleukin-6,IL-6)水平。对大鼠肝组织切片进行HE染色和油红O染色,于光学显微镜下观察。采用免疫组织化学法、蛋白质免疫印迹法及实时荧光定量聚合酶链式反应(real time polymerase chain reaction,RT-PCR)检测大鼠LXRα、SREBP-1及FAS的表达水平。结果 (1)正常组、模型组、健脾疏肝丸组和易善复组大鼠ALT[(3.40±0.81)U/L vs(9.98±2.27)U/L vs(7.80±1.52)U/L vs(6.43±1.89)U/L]、AST [(10.61±1.17)U/L vs(23.63±4.82)U/L vs(18.04±2.98)U/L vs(16.42±3.30)U/L]、TNF-α[(2.40±0.96)×10-3μg/L vs(6.64±0.92)×10-3μg/L vs(4.87±1.35)×10-3μg/L vs(4.45±1.39)×10-3μg/L]、IL-6 [(0.95±0.81)pg/ml vs(7.88±3.08)pg/ml vs(3.17±1.26)pg/ml vs(1.64±0.55)pg/ml]、TG [(0.33±0.13)mmol/L vs(0.90±0.24)mmol/L vs(0.62±0.37)mmol/L vs 0.62(0.46,0.66)mmol/L]、TC [(2.10±0.42)mmol/L vs 5.34(5.17,6.12)mmol/Lvs(3.68±0.63)mmol/Lvs(3.41±0.81)mmol/L]、HDL-C [(1.07±0.17)mmol/Lvs(0.62±0.14)mmol/Lvs(0.78±0.13)mmol/Lvs(0.79±0.12)mmol/L]及LDL-C[0.38(0.26,0.41)mmol/L vs(0.69±0.11)mmol/L vs(0.41±0.13)mmol/L vs(0.43±0.10)mmol/L]水平差异均有统计学意义(P <0.05)。与正常组相比,模型组AST、ALT、LDL-C、TG、TC、IL-6及TNF-α显著升高,HDL-C显著降低(P <0.05);与模型组相比,健脾疏肝丸组和易善复组AST、ALT、LDL-C、TG、TC、IL-6、TNF-α显著降低,HDL-C显著升高(P<0.05);与健脾疏肝丸组相比,易善复组大鼠血清TC显著降低(P <0.05),其他各指标差异无统计学意义(P> 0.05)。(2)肝组织HE染色和油红O染色示:与正常组相比,模型组大鼠肝脏脂肪变严重;与模型组相比,健脾疏肝丸组大鼠肝脏脂肪变减轻。(3)免疫组织化学结果表明,正常组、模型组、健脾疏肝丸组和易善复组大鼠LXRα(345872±52737 vs 544998±55506 vs 436319±65076 vs 448588±104641)、SREBP-1(259408±71143 vs 538701±62336vs 399705±102395 vs 394167±158047)和FAS(201683±48205 vs 466884±74934 vs 425589±63672 vs417852±84373)相对表达水平差异有统计学意义(P <0.05)。与正常组相比,模型组各蛋白相对表达水平均显著升高(P <0.05);与模型组相比,健脾疏肝丸组与易善复组各蛋白相对表达水平显著下降(P <0.05);健脾疏肝丸组与易善复组各蛋白相对表达水平差异无统计学意义(P> 0.05)。(4)Western blot结果表明,正常组、模型组、健脾疏肝丸组和易善复组大鼠LXRα[0.80±0.29 vs 1.57(1.30,1.67) vs 1.09±0.30 vs 1.10±0.36]、SREBP-1(0.42±0.12 vs 1.15±0.45 vs 0.86±0.20 vs 0.84±0.20)和FAS(0.43±0.12 vs 1.10±0.40 vs 0.81±0.26 vs 0.80±0.28)相对表达水平差异有统计学意义(P <0.05)。与正常组对比,模型组小鼠各蛋白相对表达水平显著升高(P <0.05);与模型组对比,健脾疏肝丸组及易善复组小鼠LXRα蛋白相对表达水平显著降低(P <0.05),健脾疏肝丸组与易善复组相比,各蛋白相对表达水平差异无统计学意义(P> 0.05)。(5)正常组、模型组、健脾疏肝丸组和易善复组大鼠肝组织LXRα[1.13±0.38 vs 4.14(4.01,4.35) vs 2.65±1.85 vs 1.35(0.54,4.23)]、SREBP-1、FAS[1.37±0.49 vs 4.35±1.97 vs 1.98(1.88,3.22)m RNA相对表达量差异有统计学意义(P <0.05)。与正常组相比,模型组LXRα[1.46±0.51 vs 6.13±1.17 vs 3.82±2.06 vs 1.56(1.19,4.74)]、SREBP-1、FAS vs1.83(1.64,4.29)] m RNA相对表达量显著升高(P <0.05);与模型组对比,健脾疏肝丸组及易善复组LXRα、SREBP-1、FAS m RNA表达均显著降低(P <0.05),健脾疏肝丸组与易善复组相比,LXRα、SREBP-1、FAS m RNA表达差异无统计学意义(P> 0.05)。结论健脾疏肝丸对大鼠NAFLD的改善作用可能与其抑制LXRα-SREBP-1-FAS信号转导通路有关。展开更多
文摘Soil salinity badly affects agriculture productivity through accumulation of salts in upper layers of soils. The harmful effects of salts in arable lands have influenced modern as well as ancient civilizations. A pot study was carried out to test the performance of two PGPR isolates (KS 8, KS 28) on sunflower (SMH-0917) under different salinity levels (8, 10 and 12 dS·m-1). These salinity levels were developed by adding calculated amount of salts (NaCl, Na2SO4, CaCl2 and MgSO4) with ratio of 3:4:2:1. The bacterial strains KS 8 and KS 28 were applied separately in two treatments while third treatment was co-inoculation (KS mix). Completely randomized experimental design (CRD) was used and data were collected at flowering stage about pre-decided plant growth parameters (plant height, shoot dry weight and root dry weight). The bacterial isolate KS 8 showed an increase of 26, 102% and 83% in plant height, shoot dry weight and root dry weight at EC 8 dS·m-1, while this improvement was 67%, 163% and 296% at EC 10 dS·m-1, however an increase of 100%, 74% and 382% was recorded over control respectively at EC 12 dS·m-1. Similarly isolate KS 28 exhibited an increase of 14%, 69% and 54% in plant height;shoot dry weight and root dry weight at EC 8 dS·m-1, whereas this improvement was 56%, 163% and 188% at EC 10 dS·m-1, while an increase of 60%, 41% and 282% was registered respectively over control at EC 12 dS·m-1. The increase due to mixture treatments was 4%, 41% and 16% in plant height, shoot dry weight and root dry weight at EC 8 dS·m-1, while an increase of 33%, 57% and 100% at EC 10 dS·m-1, whereas an improvement of 53%, 33% and 164% respectively was noted at EC 12 dS·m-1 over un-inoculated. The isolate KS 8 performed better than KS 28 and mixture treatment. These two PGPR strains could be used to mitigate the adverse impact caused by salinity stress on sunflower.
基金This paper was translated from its Chinese version in Chinese Journal of Rice Science.
文摘To elucidate the relationship between ethylene evolution from the grains and the appearance quality of rice, ten different rice genotypes were used to determine the ethylene evolution rate, 1-aminocylopropane-1-carboxylic acid （ACC） concentration in grains during grain filling and the appearance quality of rice, and the effects of chemical regulators on concentrations of ethylene and ACC in the grains during grain filling were also investigated to verify the roles of ethylene in the rice quality formation. The ethylene evolution rates and ACC concentrations in grains during the mid and late grain filling stages were very significantly and positively correlated with chalky kernel percentage and chalkiness. The cultivars with a low ACC concentration in grains exhibited a close amyloplast arrangement and small space between starch granules, whereas those with a high ACC concentration in grains showed a loose arrangement and wide space between the granules. Application of 1 μmol/L ACC to panicles at mid and late grain filling stages significantly loosened amyloplast arrangement and increased chalky kernel percentage, chalky area and chalkiness, and the results were reversed when 1 μmol/L amino-ethoxyvinylglycine, an inhibitor of ACC synthesis enzyme, was applied to panicles. A practice of moderate dry-wet alternate irrigation reduced ethylene evolution and ACC concentration in grains and thereby reduced chalkiness. The results suggested that ethylene and ACC in grains play an important role in the endosperm structure and appearance quality of rice, and the appearance quality would be improved by reducing ethylene evolution and ACC in grains through either variety breeding and selection, or chemical regulations or cultivation techniques.
基金The program was supported by University Science Foundation of TianjinEducational Committee (20050609) and Tianjin Normal University
文摘1-H-Pyrrole-2-carboxylic acid [2-（naphthalen-1-ylamino）-ethyl]-amide has been synthesized and characterized. Its crystal is of monoclinic, space group P2 1/n with a = 5.930（6）, b = 12.144（13）, c = 20.10（2） , A, β = 95.709（17）°, V= 1441（3） ,A, Z= 4, C17H17N3O, Mr= 279.34, Dc= 1.288 g/cm^3, F（000） = 592, μ（MoKa） = 0.083 mm^-1, S = 1.019, R = 0.0473 and wR = 0.1181 for 1713 observed reflections with I 〉 2 σ（I）. X-ray diffraction reveals that two molecules of the title compound form a dimer through a pair of N-H…O hydrogen bonds.
基金Supported by National Natural Science Foundation of ChinaNo. 82074241+1 种基金Project of Jiangsu Province Hospital of Traditional Chinese Medicine Peak TalentNo. y2021rc36
文摘BACKGROUND 3,6-dichlorobenzo[b]thiophene-2-carboxylic acid(BT2)is a benzothiophene carboxylate derivative that can suppress the catabolism of branched-chain amino acid(BCAA)-associated mammalian target of rapamycin complex 1(mTORC1)activation.Previous studies have demonstrated the therapeutic effects of BT2 on arthritis,liver cancer,and kidney injury.However,the effects of BT2 on ulcerative colitis(UC)are unknown.AIM To investigate the anti-UC effects of BT2 and the underlying mechanism.METHODS Mouse UC models were created through the administration of 3.5%dextran sodium sulfate(DSS)for 7 d.The mice in the treated groups were administered salazosulfapyridine(300 mg/kg)or BT2(20 mg/kg)orally from day 1 to day 7.At the end of the study,all of the mice were sacrificed,and colon tissues were removed for hematoxylin and eosin staining,immunoblot analyses,and immunohistochemical assays.Cytokine levels were measured by flow cytometry.The contents of BCAAs including valine,leucine,and isoleucine,in mouse serum were detected by liquid chromatography-tandem mass spectrometry,and the abundance of intestinal flora was analyzed by 16S ribosomal DNA sequencing.RESULTS Our results revealed that BT2 significantly ameliorated the inflammatory symptoms and pathological damage induced by DSS in mice.BT2 also reduced the production of the proinflammatory cytokines interleukin 6(IL-6),IL-9,and IL-2 and increased the anti-inflammatory cytokine IL-10 level.In addition,BT2 notably improved BCAA catabolism and suppressed mTORC1 activation and cyclooxygenase-2 expression in the colon tissues of UC mice.Furthermore,highthroughput sequencing revealed that BT2 restored the gut microbial abundance and diversity in mice with colitis.Compared with the DSS group,BT2 treatment increased the ratio of Firmicutes to Bacteroidetes and decreased the abundance of Enterobacteriaceae and Escherichia-Shigella.CONCLUSION Our results indicated that BT2 significantly ameliorated DSS-induced UC and that the latent mechanism involved the suppression of BCAA-associated mTORC1 activation and modulation of the intestinal flora.
基金Supported by Hong Kong Research Grant Council （HKUST6102/02M and HKUST6105/01M） and the National Natural Science Foundation of China （30129001）.
文摘1-Aminocyclopropane-1-carboxylic acid （ACC） synthase （ACS） is the principal enzyme in phytohormone ethylene biosynthesis. Previous studies have shown that the hypervariable C-terminus of ACS is proteolytically processed in vivo. However, the protease responsible for this has not yet been identified. In the present study, we investigated the processing of the 55-kDa full-length tomato ACS （LeACS2） into 52-, 50- and 49-kDa truncated isoforms in ripening tomato （Lycopersicon esculentum Mill. cv. Cooperation 903） fruit using the sodium dodecyl sulfate-boiling method. Meanwhile, an LeACS2-processing protease was purified via multi-step column chromatography from tomato fruit. Subsequent biochemical analysis of the 64-kDa purified protease revealed that it is a metalloprotease active at multiple cleavage sites within the hypervariable C-terminus of LeACS2. N-terminal sequencing and matrix-assisted laser desorption/ionization time-of-flight analysis indicated that the LeACS2-processing metalloprotease cleaves at the C-terminal sites Lys^438, Glu^447, Lys^448, Asn^456, Ser^460, Ser^462, Lys^463, and Leu^474, but does not cleave the N- terminus of LeACS2. Four C-terminus-deleted （26-50 amino acids） LeACS2 fusion proteins were overproduced and subjected to proteolysis by this metalloprotease to identify the multiple cleavage sites located on the N-terminal side of the phosphorylation site Ser^460. The results indisputably confirmed the presence of cleavage sites within the region between the α-helix domain （H14） and Ser^460 for this metalloprotease. Furthermore, the resulting C-terminally truncated LeACS2 isoforms were active enzymatically. Because this protease could produce LeACS2 isoforms in vitro similar to those detected in vivo, it is proposed that this metalloprotease may be involved in the proteolysis of LeACS2 in vivo.
基金supported by grants from National Natural Science Foundation (31361140355, 31401893, 31272184)National Key Development Plan Pilot Projects (Functional genomics research and application of wheat and other crops)+3 种基金Beijing Scholar Program and Beijing Excellent Talents Program (2014000021223TD03)Beijing Municipal Science & Technology Commission (6141001, 6144023, WRDMC01)Ministry of Science and Technology of China (2015BAD02B0202, 2014BAD01B08)Ministry of Agriculture of China (CARS-26)
文摘Summary Although it has been reported previously that ethylene plays a critical role in sex determination in cucurbit species, how the andromonoecy that carries both the male and hermaphroditic flowers is determined in watermelon is still unknown. Here we showed that the watermelon gene 1-aminocyclopropane-1-carboxylate syn- thase 4 CCitACS4）, expressed specifically in carpel primor- dia, determines the andromonoecy in watermelon. Among four single nucleotide polymorphism （SNPs） and one lnDel identified in the coding region of CitACS4, the C364W mutation located in the conserved box 6 was co- segregated with andromonoecy. Enzymatic analyses showed that the C364W mutation caused a reduced activity in CitACS4. We believe that the reduced CitACS4 activity may hamper the programmed cell death in stamen primordia, leading to the formation of hermaphroditic flowers.
文摘目的探讨健脾疏肝丸对非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)大鼠肝X受体(liver X receptorα,LXRα)-固醇调节元件结合蛋白-1(sterolregulatory element-binding protein-1,SREBP-1)-脂肪酸合成酶(fatty acid synthase,FAS)信号转导通路的影响。方法将32只无特定病原体(specific pathogen free,SPF)SD雄性大鼠按照随机数字表法分为正常组、模型组、健脾疏肝丸组和易善复组,每组8只。正常组进食普通饲料,其他组进食高脂饲料,健脾疏肝丸组给予4.86 g/(kg·d)灌胃,易善复组给予0.123 g/(kg·d)灌胃,正常组和模型组给予等量蒸馏水灌胃。灌胃与造模同时进行,共8周。检测大鼠血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、高密度脂蛋白(high-density lipoprotein cholesterol,HDL-C)、低密度脂蛋白(low-density lipoprotein cholesterol,LDL-C)、甘油三酯(triglycerides,TG)、总胆固醇(total cholesterol,TC)、肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)及白细胞介素-6(interleukin-6,IL-6)水平。对大鼠肝组织切片进行HE染色和油红O染色,于光学显微镜下观察。采用免疫组织化学法、蛋白质免疫印迹法及实时荧光定量聚合酶链式反应(real time polymerase chain reaction,RT-PCR)检测大鼠LXRα、SREBP-1及FAS的表达水平。结果 (1)正常组、模型组、健脾疏肝丸组和易善复组大鼠ALT[(3.40±0.81)U/L vs(9.98±2.27)U/L vs(7.80±1.52)U/L vs(6.43±1.89)U/L]、AST [(10.61±1.17)U/L vs(23.63±4.82)U/L vs(18.04±2.98)U/L vs(16.42±3.30)U/L]、TNF-α[(2.40±0.96)×10-3μg/L vs(6.64±0.92)×10-3μg/L vs(4.87±1.35)×10-3μg/L vs(4.45±1.39)×10-3μg/L]、IL-6 [(0.95±0.81)pg/ml vs(7.88±3.08)pg/ml vs(3.17±1.26)pg/ml vs(1.64±0.55)pg/ml]、TG [(0.33±0.13)mmol/L vs(0.90±0.24)mmol/L vs(0.62±0.37)mmol/L vs 0.62(0.46,0.66)mmol/L]、TC [(2.10±0.42)mmol/L vs 5.34(5.17,6.12)mmol/Lvs(3.68±0.63)mmol/Lvs(3.41±0.81)mmol/L]、HDL-C [(1.07±0.17)mmol/Lvs(0.62±0.14)mmol/Lvs(0.78±0.13)mmol/Lvs(0.79±0.12)mmol/L]及LDL-C[0.38(0.26,0.41)mmol/L vs(0.69±0.11)mmol/L vs(0.41±0.13)mmol/L vs(0.43±0.10)mmol/L]水平差异均有统计学意义(P <0.05)。与正常组相比,模型组AST、ALT、LDL-C、TG、TC、IL-6及TNF-α显著升高,HDL-C显著降低(P <0.05);与模型组相比,健脾疏肝丸组和易善复组AST、ALT、LDL-C、TG、TC、IL-6、TNF-α显著降低,HDL-C显著升高(P<0.05);与健脾疏肝丸组相比,易善复组大鼠血清TC显著降低(P <0.05),其他各指标差异无统计学意义(P> 0.05)。(2)肝组织HE染色和油红O染色示:与正常组相比,模型组大鼠肝脏脂肪变严重;与模型组相比,健脾疏肝丸组大鼠肝脏脂肪变减轻。(3)免疫组织化学结果表明,正常组、模型组、健脾疏肝丸组和易善复组大鼠LXRα(345872±52737 vs 544998±55506 vs 436319±65076 vs 448588±104641)、SREBP-1(259408±71143 vs 538701±62336vs 399705±102395 vs 394167±158047)和FAS(201683±48205 vs 466884±74934 vs 425589±63672 vs417852±84373)相对表达水平差异有统计学意义(P <0.05)。与正常组相比,模型组各蛋白相对表达水平均显著升高(P <0.05);与模型组相比,健脾疏肝丸组与易善复组各蛋白相对表达水平显著下降(P <0.05);健脾疏肝丸组与易善复组各蛋白相对表达水平差异无统计学意义(P> 0.05)。(4)Western blot结果表明,正常组、模型组、健脾疏肝丸组和易善复组大鼠LXRα[0.80±0.29 vs 1.57(1.30,1.67) vs 1.09±0.30 vs 1.10±0.36]、SREBP-1(0.42±0.12 vs 1.15±0.45 vs 0.86±0.20 vs 0.84±0.20)和FAS(0.43±0.12 vs 1.10±0.40 vs 0.81±0.26 vs 0.80±0.28)相对表达水平差异有统计学意义(P <0.05)。与正常组对比,模型组小鼠各蛋白相对表达水平显著升高(P <0.05);与模型组对比,健脾疏肝丸组及易善复组小鼠LXRα蛋白相对表达水平显著降低(P <0.05),健脾疏肝丸组与易善复组相比,各蛋白相对表达水平差异无统计学意义(P> 0.05)。(5)正常组、模型组、健脾疏肝丸组和易善复组大鼠肝组织LXRα[1.13±0.38 vs 4.14(4.01,4.35) vs 2.65±1.85 vs 1.35(0.54,4.23)]、SREBP-1、FAS[1.37±0.49 vs 4.35±1.97 vs 1.98(1.88,3.22)m RNA相对表达量差异有统计学意义(P <0.05)。与正常组相比,模型组LXRα[1.46±0.51 vs 6.13±1.17 vs 3.82±2.06 vs 1.56(1.19,4.74)]、SREBP-1、FAS vs1.83(1.64,4.29)] m RNA相对表达量显著升高(P <0.05);与模型组对比,健脾疏肝丸组及易善复组LXRα、SREBP-1、FAS m RNA表达均显著降低(P <0.05),健脾疏肝丸组与易善复组相比,LXRα、SREBP-1、FAS m RNA表达差异无统计学意义(P> 0.05)。结论健脾疏肝丸对大鼠NAFLD的改善作用可能与其抑制LXRα-SREBP-1-FAS信号转导通路有关。
