小兴安岭阔叶林沼泽土壤CO_2、CH_4和N_2O排放规律及其影响因子

采用野外静态箱-气相色谱法,研究了小兴安岭典型阔叶林沼泽生长季节土壤CO2、CH4和N2O排放季节变化规律、源/汇功能及主要影响因子。结果表明:①苔草沼泽、毛赤杨沼泽和白桦沼泽生长季节土壤CO2、CH4、N2O排放分别集中在夏季、夏秋季、春夏季,平均排放通量依次为487.89、382.27、514.63 mg.m-2.h-1,1.88、1.03、0.04 mg.m-2.h-1,3.70、58.61、11.73μg.m-2.h-1。②三者生长季节土壤CO2排放通量与气温和0-20 cm土壤温度均呈显著正相关;苔草沼泽CH4排放通量与30-40cm土壤温度呈显著正相关,毛赤杨沼泽CH4排放通量与地表温度呈显著负相关;白桦沼泽N2O排放通量与地表温度呈显著正相关。苔草沼泽N2O排放与水位呈显著负相关;毛赤杨沼泽CH4排放与水位呈显著正相关;白桦沼泽CO2排放与水位呈显著负相关。③三者生长季节土壤均为CO2、CH4、N2O排放源(17.56、13.76、18.53 t.hm-2;67.54、37.05、1.30 kg.hm-2;0.13、2.11、0.42 kg.hm-2),三者CO2排放量相近(5.5%-21.6%);苔草沼泽为CH4的强排放源,毛赤杨沼泽为中排放源,白桦沼泽为弱排放源;毛赤杨沼泽为N2O的强排放源,白桦沼泽为中排放源,苔草沼泽为弱排放源。     

火干扰对小兴安岭毛赤杨沼泽温室气体排放动态影响及其影响因素

利用静态箱-气相色谱法,对小兴安岭轻度火烧毛赤杨沼泽CH4、CO2、N2O生长季排放通量进行研究.结果表明:火烧使毛赤杨沼泽生长季CH4排放通量提高485.2％,CO2和N2O排放通量分别下降45.5％、24.8％.火烧未改变CH4季节性排放规律,但改变了CO2、N2O季节性变化规律.火烧样地CH4排放通量与土壤15cm温度间存在显著正相关性关系而与水位相关性不显著,火烧样地CO2排放与土壤0-30 cm温度呈显著或极显著正相关,与水位极显著负相关.对照样地CO2排放通量与土壤0-15 cm温度呈显著或极显著正相关,与水位极显著负相关,火烧使毛赤杨沼泽CH4排放源的强度增强,CO2、N2O的排放消弱,全球温室潜势下降约为43.34％.     

小兴安岭典型苔草和灌木沼泽N2O排放及影响因子

为了研究小兴安岭林区典型苔草和灌木沼泽N2O排放通量的季节动态、年际动态及其与环境因子的关系,并估算排放总量,2007和2008年在植物生长季采用静态箱-气相色谱法,对小兴安岭林区典型修氏苔草（Carex schmidtii）沼泽和油桦-修氏苔草（Betula ovalifolia-Carex schmidtii）灌木沼泽N2O排放进行了监测。结果表明,苔草和灌木沼泽2007年生长季N2O排放总量分别为0.14和0.29 kg/hm2;2008年分别为0.68和-0.10 kg/hm2。苔草和灌木沼泽N2O排放通量除灌木沼泽2008年变化规律性不明显外,均具有比较显著的季节变化,最大排放出现在夏季或夏、秋季节,其中2007年N2O排放平均通量为0.0037和0.0082 mg?m-2?h-1;2008年为0.016和-0.0025 mg?m-2?h-1。分析表明,苔草沼泽N2O排放年际差异不显著,灌木沼泽N2O排放年际差异显著;不同类型沼泽间N2O排放差异不显著;仅苔草沼泽2007年N2O排放通量与水位具有显著的负相关性（r=-0.52,P < 0.05,n=15）。     

施氮对桉树人工林生长季和非生长季土壤温室气体通量的影响

森林在调控温室气体排放方面有重要作用,随着人工林的迅速发展,其温室气体通量和对施肥的响应逐渐引起广泛关注。为了解施氮对桉树人工林生长季和非生长季土壤温室气体通量的影响,在广西东门林场尾巨桉人工林样地设置低(84.2 kg N·hm-2)、中(166.8 kg N·hm-2)、高(333.7 kg N·hm-2)3个施氮水平和不施氮对照,采用静态箱-气相色谱法监测土壤CO2、N2O和CH4通量。结果表明:(1)不同施氮处理的桉树人工林土壤CO2、CH4和N2O年均排放通量分别为214~271 mg CO2·m-2·h-1、-47~-37 kg CH4·m-2·h-1和16~203 kg N2O·m-2·h-1;土壤CO2排放通量在生长季高于非生长季,CH4和N2O通量未表现出明显季节变化。(2)施氮显著增加了土壤CO2和N2O年均排放通量,其促进效应主要集中在生长季(施氮后的4个月,即6—9月),且随时间增加,效应减弱。(3)施氮显著降低了土壤CH4年均吸收通量。因此,在维持桉树人工林生产力的基础上,结合季节变化,合理调控施氮量将有助于减少桉树林土壤温室气体排放。     

火干扰对小兴安岭草丛、灌丛沼泽温室气体短期排放的影响

利用静态箱-气相色谱法,研究了火烧干扰对小兴安岭草丛、灌丛沼泽生长季CH4、CO2、N2O排放的季节变化及影响因子结果表明:火干扰使草丛、灌丛沼泽生长季的平均气温和各层土壤温度提高0.1—2.0℃,水位平均下降2.7 cm。火干扰使草丛、灌丛沼泽样地CH4排放通量提高了56%、524.9%,CO2排放通量分别下降了57.3%、14.5%,N2O排放通量分别下降27.1%,64.9%。火烧前后草丛沼泽CH4、N2O与灌丛沼泽CO2排放通量季节性规律未发生变化。火干扰改变了草丛沼泽生长季CO2、灌丛沼泽N2O排放通量的季节性变化规律。草丛沼泽对照样地CH4排放通量与5 cm土壤温度存在显著相关性,草丛沼泽CH4排放通量与水位相关性不显著。灌丛沼泽CH4排放通量与各层土壤温度及水位均无显著相关性。草丛、灌丛沼泽对照样地土壤CO2排放通量与0—15 cm土壤温度呈显著或极显著正相关,火烧样地与0—30 cm土壤温度呈显著或极显著正相关。草丛、灌丛沼泽对照、火烧样地土壤CO2排放通量与水位极显著负相关。火干扰使草丛、灌丛沼泽CH4排放源的强度增强,CO2、N2O的排放消弱,全球温室潜势下降约为23.3%。火干扰能够减少草丛、灌丛沼泽温室气体排放。     

干旱年份三江平原沼泽甲烷(CH4)排放及影响因子

为了研究淡水沼泽CH4排放的影响因子及其机制,在中国东北部的三江平原样地观测了CH4排放通量以及部分影响因子.结果表明,在2003年4～10月生长季内,毛果苔草沼泽CH4通量平均值为3.30 mg·m-2·h-1(最小值为0.65 mg·m-2·h-1,最大值为13.47 mg·m-2·h-1),低于小叶章沼泽化草甸CH4通量平均值4.91 mg·m-2·h-1(最小值为-0.12 mg·m-2·h-1,最大值为16.25 mg·m-2·h-1).在不同阶段,两种类型沼泽CH4通量有明显的变化,控制因子也不一样.4月份低温限制了CH4通量,两种类型沼泽差异很小.5～7月下旬,小叶章沼泽化草甸CH4通量显著高于毛果苔草沼泽CH4通量(P＜0.01),这种差异是由融冻作用造成的.8月份,植株密度和剖面中活性有机碳的浓度成为主要的影响因子,毛果苔草沼泽CH4通量比小叶章沼泽化草甸高.     

小兴安岭典型草丛沼泽湿地CO2、CH4和N2O的排放动态及其影响因素

2007年6～10月,采用静态箱-气相色谱法,同步研究了小兴安岭典型修氏苔草(Carex schmidtii)沼泽湿地CO2、CH4和N2O排放通量的季节动态及其与环境因子的关系,估算CO2、CH4和N2O的生长季排放量,探讨了沼泽湿地碳与氮的源汇关系.结果表明:草丛沼泽生长季节温室气体排放量以CO2占绝对优势(99.61%),CH4的排放量次之(0.39%),N2O的排放量最低(0.000 7%),且为碳、氮的吸收汇(分别为固定量的53.93%和0.04%);CO2、CH4和N2O生长季平均排放通量依次为487.89、1.88和0.004 mg·m-2·h-1,且具有明显的季节变化特征,CO2和N2O的最高排放量均出现在夏季(6月24日至8月14日和7月14日至8月14日),CH4的最高排放量出现在夏秋季(8月24日至9月24日),其中,CO2季节变化与空气温度和0～20 cm土壤温度具有显著相关性(p＜0.05),CH4与空气温度具有显著相关性(p<0.01),N2O与水位具有显著的负相关性(p＜0.05).     

若尔盖高原沼泽湿地N_2O排放通量研究

应用静态箱/气相色谱法,测定了若尔盖高原沼泽N2O排放能量,测定期为该地植物生长期,即2004年4 月末至10月初。结果表明,若尔盖高原沼泽湿地N2O排放通量平均值为0.010 mg·m-2h-1,最大值为0.079 mg·m-2h-1, 最小值为-0.051mg·m-2h-1。高峰排放期为5月,最低排放期为地表水深最大的6月。沼泽湿地N2O排放通量季节变化与沼泽湿地水深呈负相关关系。沼泽湿地N2O排放通量日变化与大气温度呈正相关关系,排放高值出现在午后。若尔盖高原沼泽湿地在植物生长期的年排放总量约为0.159Gg·a-1。     

玉渡山水库生长季温室气体排放特征及其影响因素

为了探讨温带水库温室气体排放规律,采用静态箱-色谱分析法,研究了温带地区库龄10年内的北京玉渡山水库生长季3种温室气体CO2、CH4及N2O排放特征,及其影响因子。结果表明:样地类型、测定月份与样地类型交互作用对3种温室气体通量影响极显著,5月消落带CO2通量(664.31mg·m-2·h-1)达到最大,显著高于入库口和浅水区;8月消落带CH4通量(0.87mg·m-2·h-1)及N2O通量(3.05mg·m-2·h-1)最大;8月,切除消落带样地地上植物后,3种温室气体通量均有所降低。CO2通量与地下5cm地温、氧化还原电位和水体总氮显著正相关,与地上生物量和水体pH显著负相关;CH4通量与地表温度、地上生物量、水体pH呈显著相关,与水体总氮和水体铵态氮显著负相关;N2O通量与水体总氮含量显著相关,与水体pH显著负相关。采取平均估值法初步推测,在生长季,水库消落带、入库口及浅水区CO2排放量依次为15960、2160、-70kg·hm-2;CH4排放量依次20.04、-7.05、14.8kg·hm-2;N2O排放量依次83.42、3.79、-1.54kg·hm-2;表明消落带3种温室气体的排放量均较高,为玉渡山水库3种温室气体排放的重点区域。     

青藏高原高寒草甸非生长季温室气体排放特征及其年度贡献

高寒草甸是青藏高原地区的主要植被类型,目前对其温室气体研究多集中于生长季.本文利用静态箱-气相色谱法,对非生长季高寒草甸温室气体排放特征及其与主要环境因子的关系进行了研究.结果表明:非生长季高寒草甸表现为CO2和N2O的源、CH4的汇.其中非生长季CO2通量平均值为89.33 mg·m-2·h-1,累积排放通量为280.01g· m-2;CH4通量平均值为-11.35 μg·m-2·h-1,累积吸收通量为124.74 mg·m-2;N2O通量平均值为8.02 μg·m-2·h-1,累积排放通量为39.51 mg·m-2.非生长季CO2、CH4和N2O累积排放通量分别占全年的13.33％、53.47％和62.67％.冻融期(2012年4月)CH4累积吸收通量较小,只占非生长季的4.5％;而CO2和N2O累积排放通量较大,分别占非生长季的25.8％和20.8％.非生长季CO2通量与温度(气温、5和10 cm土壤温度)和5 cm土壤湿度均存在显著正相关关系,而CH4和N2O通量仅与5 cm土壤湿度存在显著正相关.研究表明,虽然冻融期CH4累积吸收通量在非生长季累积量中比重较小,但非生长季CH4和N2O累积排放量却占全年累积排放量的1/2以上,在温室气体累积通量评估中不容忽视.     

Inhibition of thymidylate synthase by 2′,2′-difluoro-2′-deoxycytidine (Gemcitabine) and its metabolite 2′,2′-difluoro-2′-deoxyuridine

2′,2′-Difluoro-2′-deoxycytidine (dFdC, gemcitabine) is a cytidine analogue active against several solid tumor types, such as ovarian, pancreatic and non-small cell lung cancer. The compound has a complex mechanism of action. Because of the structural similarity of one metabolite of dFdC, dFdUMP, with the natural substrate for thymidylate synthase (TS) dUMP, we investigated whether dFdC and its deamination product 2′,2′-difluoro-2′-deoxyuridine (dFdU) would inhibit TS. This study was performed using two solid tumor cell lines: the human ovarian carcinoma cell line A2780 and its dFdC-resistant variant AG6000. The specific TS inhibitor Raltitrexed (RTX) was included as a positive control. Using the in situ TS activity assay measuring the intracellular conversion of [5-³H]-2′-deoxyuridine or [5-³H]-2′-deoxycytidine to dTMP and tritiated water, it was observed that dFdC and dFdU inhibited TS. In A2780 cells after a 4 h exposure to 1 μM dFdC tritium release was inhibited by 50% but did not increase after 24 h, Inhibition was also observed following dFdU at 100 μM. No effect was observed in the dFdC-resistant cell line AG6000; in this cell line only RTX had an inhibitory effect on TS activity. In the A2780 cell line RTX inhibited TS in a time dependent manner. In addition, DNA specific compounds such as 2′-C-cyano-2′-deoxy-1-beta-D-arabino-pentafuranosylcytosine and aphidicoline were utilized to exclude DNA inhibition mediated down regulation of the thymidine kinase.Inhibition of the enzyme resulted in a relative increase of mis-incorporation of [5-³H]-2′-deoxyuridine into DNA. In an attempt to elucidate the mechanism of in situ TS inhibition the ternary complex formation and possible inhibition in cellular extracts of A2780 cells, before and after exposure to dFdC, were determined. With the applied methods no proof for formation of a stable complex was found. In simultaneously performed experiments with 5FU such a complex formation could be demonstrated. However, using purified TS it was demonstrated that dFdUMP and not dFdCMP competitively inhibited TS with a Ki of 130 μM, without ternary complex formation. In conclusion, in this paper we reveal a new target of dFdC: thymidylate synthase.     

A concise synthesis of 4-nitrophenyl 2-azido-2-deoxy- and 2-acetamido-2-deoxy-D-mannopyranosides

4-nitrophenyl 3,4,6-tri-O-acetyl-2-azido-2-deoxy-alpha- and beta-D-mannopyranosides were prepared from methyl 4,6-O-benzylidene-alpha-D-glucopyranoside and 1,3,4,6-tetra-O-acetyl-alpha-D-glucopyranose, respectively. Chemoselective reduction of both azides with hydrogen sulfide readily afforded 4-nitrophenyl 2-acetamido-4,6-di-O-acetyl-2-deoxy-alpha-D- and -beta-D-mannopyranosides in higher yields than reduction with triphenylphosphine or a polymer-supported triarylphosphine. Subsequent de-O-acetylation yielded 4-nitrophenyl 2-acetamido-2-deoxy-alpha-D-mannopyranoside and 4-nitrophenyl 2-acetamido-2-deoxy-beta-D-mannopyranoside in 20% and 44% overall yields, respectively.     

Ca2+位于H2O2上游参与H2S诱导的拟南芥气孔关闭过程

以拟南芥为材料,利用药理学实验,结合分光光度法和激光共聚焦显微技术,研究了Ca2+在硫化氢(H2S)诱导拟南芥气孔关闭过程中的作用及其与过氧化氢(H2O2)的关系。结果表明： H2S诱导气孔关闭, Ca2+螯合剂EGTA和质膜Ca2+通道阻断剂硝苯地平(Nif)能不同程度抑制H2S诱导的气孔关闭,而内质网钙泵阻断剂毒胡萝卜素(Thaps)对H2S的作用无显著影响。由此推测, Ca2+参与调节H2S诱导的拟南芥气孔关闭过程,且胞质中Ca2+来源于胞外Ca2+的内流。另外, H2S诱导拟南芥叶片NADPH氧化酶基因AtRBOHD和AtRBOHF以及细胞壁过氧化物酶基因AtPRX34表达增强,促进叶片和保卫细胞中H2O2积累, EGTA对此起抑制作用,而外源CaCl2处理上调AtRBOHD、AtRBOHF和AtPRX34的表达。表明Ca2+可能位于H2O2上游参与H2S诱导的拟南芥气孔关闭过程。     

Smurf2 is a TRAF2 binding protein that triggers TNF-R2 ubiquitination and TNF-R2-induced JNK activation

TRAF2 plays a central role in TNF-induced signalling to NF-κB and JNK/p38 MAPK. To better understand the molecular mechanisms that mediate this dual function of TRAF2, we performed a yeast two-hybrid screening for TRAF2 interacting proteins using the Sos recruitment system. This resulted in the identification of the E3 ubiquitin ligase Smurf2 as a TRAF2 binding protein. TRAF2 overexpression was shown to trigger Smurf2 ubiquitination and the formation of a TNF-R2/Smurf2 complex. Smurf2 on its turn promoted TNF-R2 ubiquitination and the relocalization of TNF-R2 as well as TRAF2 to a detergent-insoluble cell fraction. This was associated with enhanced TNF-R2-induced JNK activation, whereas TNF-R2-induced NF-κB activation remained unaffected. These results suggest an important role for Smurf2 binding to TRAF2 in determining specific signalling outputs of TNF-R2.     

Synthesis of [2S-2-H]- and [2R-2-H]hexadecanoic acids

[2S-2-²H]- and [2R-2-²H]hexadecanoic acids were synthesized in overall yields of 59–67%. Methyl(2R)-2-hydroxyhexadecanoate, from the acid produced by Hansenula sydowiorum, was converted to the p-toluenesulphonate, reduced to trideutero alcohol with lithium aluminium deuteride and oxidized to [2S-2-²H]hexadecanoic acid. Methyl (2S)-2-chlorohexadecanoate, which was a by-product of tosylation and was also prepared by chlorinatioon of the hydroxy ester with thionyl chloride, on reduction and oxidation as before gave [2R-2-²H]-hexadecanoic acid. Intermediates were fully characterized, isotopic purity was 97% and optical purity was maintained throughout the syntheses. Attempts to reduce the tosyl or chloro groups, only, with sodium borodeuteride gave low yields probably due to preferential reduction of the ester group; 1,2-epoxyhexadecane was obtained from the tosylate and 2-chlorohexadecan-1-ol from the chloro ester.     

拟南芥血红蛋白1(AtGLB1)与过氧化氢的相互作用

拟南芥的血红蛋白1(AtGLB1)属于非共生的血红蛋白。在低氧胁迫中对植物细胞中过氧化氢(H2O2)内稳态的维持起了很重要的作用。为了检测AtGLB1与H2O2能否直接相互作用,我们扩增了拟南芥的AtGLB1基因,并将其克隆到原核表达质粒pET32a中,测序鉴定正确后转化大肠杆菌BL21。IPTG诱导目的蛋白表达后,镍离子亲和层析柱(Ni2+-NTA)纯化了靶蛋白。体外表达的氧合的AtGLB1能与H2O2直接相互作用。因此,与H2O2反应可能是AtGLB1清除低氧胁迫下产生的H2O2的一种方式。     

过氧化氢和氯化钙对香蕉幼苗抗寒性的影响

用H₂O₂和CaCl₂单独或混合使用的方法喷洒香蕉幼苗,并置于低温培养箱中进行冷胁迫处理,发现它们可提高香蕉幼苗冷胁迫期间叶片POD活性,降低细胞质泄漏,增加可溶性糖含量及减缓叶绿素降解,从而减轻冷伤害程度。H₂O₂和CaCl₂混合处理的效果优于单独处理,二者有协同效应。     

PPP2R2A在乳腺癌细胞中结合GFPT2并导致其去磷酸化

PPP2R2A是PP2A磷酸酶的调控亚基之一,以往的研究报道显示,PPP2R2A可促进肿瘤细胞生存和生长。本研究通过串联亲和纯化联合HPLC-Chip-ESI/MS/MS筛选PPP2R2A的相互作用蛋白质,分析结果显示,L-谷氨酰胺-D-果糖-6-磷酸转氨酶1(Glutamine-fructose-6-phosphate transaminase 1,GFPT1)和L-谷氨酰胺-D-果糖-6-磷酸转氨酶2(Glutamine-fructose-6-phosphate transaminase 2,GFPT2)是PPP2R2A可能的结合蛋白。通过免疫荧光共定位、GST Pull-down和免疫共沉淀等方法,进一步确认了PPP2R2A和GFPT1及GFPT2的相互结合。通过shRNA下调PPP2R2A后,GFPT2的磷酸化水平显著增加,但GFPT1的磷酸化水平改变不明显。GFPT2是O-GlcNAC糖基化修饰通路中的一个限速酶,在乳腺癌细胞MDA-MB-231中下调PPP2R2A后,蛋白质O-GlcNAC糖基化修饰水平增加。这些结果表明,PPP2R2A可直接结合GFPT2,并导致其去磷酸化,进而影响细胞内O-GlcNAC糖基化修饰。