盐胁迫对黄瓜幼苗根系脯氨酸和多胺代谢的影响

以2个黄瓜品种‘长春密刺’(抗盐性较强)和‘津春2号’(抗盐性较弱)为材料,采用营养液栽培,研究了盐胁迫对幼苗根系脯氨酸(Pro)和多胺(PAs)代谢的影响。结果表明,盐胁迫能提高黄瓜幼苗根系吡咯啉-5-羧酸合成酶(P5CS)活性,抑制脯氨酸脱氢酶(ProDH)活性,从而显著增加Pro含量,且‘长春密刺’变化幅度显著大于‘津春2号’;盐胁迫下,‘长春密刺’根系精氨酸脱羧酶(ADC)、鸟氨酸脱羧酶(ODC)和S-腺苷蛋氨酸脱羧酶(SAMDC)活性升高幅度显著大于‘津春2号’,而多胺氧化酶(PAO)活性升高幅度显著低于‘津春2号’,引起其根系内亚精胺(Spd)和精胺(Spm)含量显著增加;盐胁迫下,2品种根系腐胺(Put)含量呈先上升后下降的变化趋势,随着Put积累降低,Pro含量显著增加。可见,盐胁迫诱导根系较高的Pro、Spd和Spm积累有利于提高黄瓜幼苗对盐胁迫逆境的适应能力,盐胁迫下PAs代谢和Pro代谢密切相关,Put的积累一定程度上促进了Pro含量的增加。     

盐碱混合胁迫对裸燕麦幼苗叶片脯氨酸和多胺代谢的影响

为探讨裸燕麦对盐碱混合胁迫的脯氨酸(Pro)和多胺(PAs)响应机制,采用温室砂培试验,研究了低(25 mmol·L-1)、高(75 mmol·L~(-1))浓度盐碱混合胁迫0、1、3、5和7 d幼苗叶片Pro和PAs含量及相关代谢酶活性的变化。结果表明:与对照相比,盐碱混合胁迫在整个胁迫期间提高了裸燕麦叶片Pro含量,高浓度胁迫提高的幅度大于低浓度胁迫;低浓度胁迫下吡咯啉-5-羧酸合成酶(P5CS)活性提高,而鸟氨酸-δ-氨基转移酶(δ-OAT)和脯氨酸脱氢酶(Pro DH)活性受抑;高浓度胁迫下δ-OAT活性提高,而P5CS和Pro DH活性下降,说明低浓度胁迫下Pro合成以谷氨酸途径为主,高浓度胁迫下以鸟氨酸途径为主;低、高浓度盐碱混合胁迫下,游离态腐胺(Put)、亚精胺(Spd)和精胺(Spm)及结合态Spd和束缚态Spd、Spm含量提高,结合态Put、Spm和束缚态Put含量在胁迫1~5 d时提高,胁迫7 d时下降,且高浓度胁迫的变幅大于低浓度胁迫;Put/PAs和(c PAs+b PAs)/f PAs在胁迫1~3 d时提高,胁迫5~7 d时下降或变化不明显,(Spd+Spm)/Put的变化却相反;整个胁迫期间低浓度胁迫保持了较低的Put/PAs和较高的(Spd+Spm)/Put,(c PAs+b PAs)/f PAs除胁迫1 d时低浓度胁迫低于高浓度胁迫外,其余胁迫时间两浓度处理间差异不显著;精氨酸脱羧酶(ADC)和鸟氨酸脱羧酶(ODC)活性在低、高浓度盐碱混合胁迫期间提高,二胺氧化酶(DAO)活性在胁迫1~3 d下降或变化不大,胁迫5~7 d时升高,多胺氧化酶(PAO)活性的变化与DAO相反,且高浓度胁迫下ADC、ODC、DAO和PAO的变幅大于低浓度胁迫。说明盐碱混合胁迫初期(1~3 d)游离态PAs(f PAs)向结合态和束缚态PAs(cPAs+bPAs)转化增强,低浓度胁迫后期(5~7 d)Put向Spd和Spm转化的能力强于高浓度胁迫,但高浓度胁迫在整个胁迫期间PAs合成和分解的速率更高。     

外源脯氨酸对盐胁迫下甜瓜脯氨酸代谢的影响

为探明外源脯氨酸对盐胁迫下甜瓜脯氨酸代谢的影响,以甜瓜品种‘雪美’为材料采用营养液栽培,对盐胁迫(100mmol·L-1 NaCl)、盐胁迫下添加外源脯氨酸(100mmol·L-1 NaCl+0.2mmol·L-1 Proline)以及对照3种处理后甜瓜幼苗叶片脯氨酸(Pro)含量、吡咯啉-5-羧酸合成酶(P5CS)、鸟氨酸转氨酶(OAT)和脯氨酸脱氢酶(ProDH)活性进行测定,并对OAT和ProDH基因进行克隆及半定量表达分析。结果显示:与对照相比较,盐胁迫条件下甜瓜幼苗叶片内Pro含量显著增加,P5CS活性增幅大于OAT活性,OAT基因表达量大部分时段内没有增加,ProDH活性下降,ProDH基因表达量减少;盐胁迫下添加外源脯氨酸进一步使幼苗叶片内Pro含量增加、OAT、ProDH活性提高、P5CS活性降低,并且使OAT基因表达量迅速增加、ProDH基因表达量先增加后回落。研究表明,盐胁迫条件下,甜瓜幼苗体内脯氨酸积累主要是通过增强脯氨酸的谷氨酸合成途径和抑制脯氨酸降解来实现;适量外源脯氨酸可以增强盐胁迫幼苗脯氨酸的鸟氨酸合成途径,但对谷氨酸合成途径有一定的抑制作用;通过调节合成和降解2种代谢途径进一步提高了脯氨酸含量,从而增强甜瓜幼苗耐盐胁迫能力。     

NaCl胁迫下海马齿(Sesuvium portulacastrum L.)植株内游离脯氨酸的合成积累途径

以海马齿(Sesuvium portulacastrum L.)为材料,测定了在6个NaCl胁迫水平上(0、0.2、0.4、0.6、0.8、1.0mol· L-1)的植株内游离脯氨酸的含量及其δ-OAT、PSCS、POD、SOD等酶的活性.结果表明:在NaCl胁迫下,海马齿植株内游离脯氨酸积累量比CK显著地增多了14.5％～33.5％,δ-OAT和P5CS活性分别比对照(CK)增强3.1％～10.1％和26.7％ ～46.1％,因此,在NaCl胁迫下海马齿植株内游离脯氨酸合成积累的两个途径均被启动和发生合成积累作用,并表现出以Glu→Pro途径为主,Orn→Pro途径为辅;在5个NaCl胁迫水平上,海马齿植株内游离脯氨酸有显著的积累效应,在0.6 mol·L-1处理水平时达到积累量的高峰值.同时,δ-OAT、P5CS、POD、SOD等酶的活性呈现出与游离脯氨酸积累量有大致相同的变化趋势.     

外源ABA对低温胁迫下小桐子幼苗脯氨酸积累及其代谢途径的影响

本文以小桐子为材料,研究了外源脱落酸（ABA）对低温胁迫下小桐子幼苗脯氨酸积累的影响。结果表明,低温胁迫（5℃）可促进小桐子幼苗体内脯氨酸的积累,上调脯氨酸合成关键酶Δ¹-吡咯琳-5-羧酸合成酶（P5CS）和鸟氨酸转氨酶（OAT）的活性,上调P5CS基因（JcP5CS）的表达,及抑制脯氨酸降解酶脯氨酸脱氢酶（Pro DH）的活性。用外源ABA处理低温胁迫下的小桐子幼苗,发现150μmol·L^-1的ABA可显著提高其低温胁迫下的脯氨酸含量,上调P5CS的活性和JcP5CS的表达水平,及抑制ProDH的活性。表明外源ABA可通过活化脯氨酸合成的谷氨酸途径和抑制脯氨酸的降解途径来促进低温胁迫下小桐子幼苗脯氨酸的积累。     

外源亚精胺对盐胁迫下毕氏海蓬子体内多胺含量的影响

用含不同浓度(0、100、200、300、400和500 mmol·L 1)NaCl的Hoagland营养液及叶面喷施0.1 mmol·L-1外源亚精胺(Spd)处理毕氏海蓬子幼苗,研究外源Spd对NaCl胁迫下海蓬子体内游离态、结合态和束缚态3种形态多胺含量的影响,分析内源多胺含量的变化与植物耐盐性的关系.结果表明:(1)随盐胁迫浓度的升高,海蓬子幼苗叶片中3种形态腐胺(Put)含量均先降后升;同期的游离态Spd含量持续上升,结合态和束缚态Spd含量均先升后降;同期的游离和结合态精胺(Spm)含量均先升后降,而其束缚态Spm含量呈上升趋势;游离态和束缚态多胺(PAs)总量变化随盐浓度升高均呈上升趋势,而结合态PAs总量先升后降.(2)3种形态(Spd+ Spm) /Put比值均先升后降,而3种形态Put/PAs比值则均呈先降后升的相反趋势.(3)外源Spd处理提高了海蓬子幼苗叶片中结合态和束缚态PAs总量,也提高了游离态和束缚态(Spd+Spm)/Put比值.研究发现,外源Spd参与了NaCl胁迫下海蓬子内源PAs代谢的调节,可能通过促进盐胁迫植株中Put向Spd和Spm的转化,以及游离态多胺向结合态和束缚态多胺的转化来增强海蓬子耐盐性.     

一氧化氮参与调节盐胁迫下脱落酸诱导的小麦幼苗叶片脯氨酸的累积

不同浓度(0.01～5.00mmol/L)的外源一氧化氮(NO)供体硝普钠(SNP)以浓度依赖性的性式诱导150mmol/L NaCl胁迫下小麦(Triticum aestivum L.cv.Yangmai 158)幼苗叶片脯氨酸的累积。其中0.1 mmol/L的SNP效果最明显,而结合采用NO清除剂c-PTIO和血红蛋白的处理均分别逆转了该效应。研究结果还发现:0.1 mmol/L SNP诱导的脯氨酸累积还可能有利于盐胁迫下小麦幼苗的保水性;0.1 mmol/L的SNP显著激活了内源ABA的合成,而结合血红蛋白的处理则证实,在外源ABA诱导脯氨酸累积的过程中NO可能作用于ABA信号分子的下游,但NO和ABA信号分子在此诱导反应中不存在累积效应。进一步研究脯氨酸合成和降解的酶促反应途径,发现外源NO处理前4天内可能主要是通过提高Δ~1-吡咯啉-5-羧酸合成酶(P5CS)的活性来促进脯氨酸的合成,以后直至第8天主要是通过抑制脯氨酸脱氢酶(ProDH)的活性来抑制脯氨酸的降解;ABA对于P5CS和ProDH活性的调节能力弱于NO。此外,Ca~(2 )在NO诱导的盐胁迫下小麦叶片脯氨酸累积的信号分子途径中起重要的介导作用。     

钙对低氧胁迫下黄瓜幼苗体内多胺及多胺氧化酶的影响

以‘中农8号’黄瓜品种为实验材料,采用营养液栽培法研究了钙对根际低氧胁迫下黄瓜幼苗体内多胺(PAs)含量及多胺氧化酶(PAO)活性的影响。结果表明:(1)各处理黄瓜幼苗根系和叶片中的PAs含量以及3种形态的腐胺(Put)、亚精胺(Spd)、精胺(Spm)含量均表现为低氧高钙(8 mmol.L-1Ca2 )>低氧常钙(2 mmol.L-1Ca2 )>低氧缺钙(0 mmol.L-1Ca2 )>通气常钙(2 mmol.L-1Ca2 )处理,而PAO活性却表现出相反的趋势(通气常钙>低氧缺钙>低氧常钙>低氧高钙),且处理间大多存在显著差异(P<0.05);根系中的PAs含量明显高于叶片,而PAO活性明显低于叶片。(2)黄瓜幼苗体内3种形态的PAs以游离态含量最高,其次是结合态,最低为束缚态;游离态和结合态PAs在叶片中均以Spd为主,在根系中均以Put为主,束缚态PAs含量在根系和叶片中均为Spd>Put>Spm。研究表明,在低氧胁迫下,营养液加钙引起黄瓜幼苗体内多胺含量的上升和PAO活性下降,钙参与了黄瓜幼苗体内多胺的代谢过程,对缓解低氧胁迫有重要作用。     

NaCl和Na2CO3对盐地碱蓬胁迫效应的比较

在相同的Na 浓度(如100 mmol/L)下,NaCl处理促进碱蓬植株干重增加,提高根系活力,而Na2CO3处理导致植株干重减少,根系活力降低;与NaCl胁迫相比,Na2CO3胁迫下叶片内Na 含量上升和K 含量下降幅度更大,叶肉细胞质Na 含量和叶内脯氨酸含量增加幅度更大,而V-H -ATPase(液泡膜H -ATPase)和V-H -PPase (液泡膜H -PPase)增加幅度较少;与NaCl胁迫不同,Na2CO3胁迫下SOD(超氧化物歧化酶)活性不是增加,而是降低,与此相一致,MDA(丙二醛)含量大幅度增加.上述结果表明,碱蓬对Na2CO3胁迫的抗性低于对NaCl的抗性,这可能与Na2CO3胁迫引起的Na 、K 离子严重失衡、活性氧清除能力降低有关.     

氯化钠胁迫下野生和栽培大豆幼苗体内的多胺水平变化

以通用的较耐盐的栽培大豆Lee68品种和对盐敏感的野生大豆N23232种群为参照,研究了盐胁迫下耐盐野生大豆BB52种群幼苗体内多胺(PAs)组分、含量及多胺氧化酶(PAO)活性的变化。结果表明,盐胁迫下BB52幼苗根PAs中Put和Spm含量下降较Lee68和N23232显著,但Spd含量下降较少．BB52叶片PAs中Put含量下降,Spd上升,(Spd+Spin)／Put值增加和Put／PAs值降低幅度与耐盐性呈正相关趋势．盐胁迫下,各材料根和叶中PAO活性上升,N23232上升最明显．探讨了多胺水平与BB52耐盐性的关系。     

壳聚糖酶的克隆表达与壳寡糖的制备分析

目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。     

国内外蝗害治理技术现状与展望

本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...     

Kinetics and thermodynamics of peroxidase- and laccase-catalyzed oxidation of N-substituted phenothiazines and phenoxazines

N -substituted phenothiazines (PTs) and phenoxazines (POs) catalyzed by fungal Coprinus cinereus peroxidase and Polyporus pinsitus laccase were investigated at pH 4–10. In the case of peroxidase, an apparent bimolecular rate constant (expressed as k _cat/K _m) varied from 1 ×10⁷M⁻¹s⁻¹to 2.6×10⁸M⁻¹s⁻¹ at pH 7.0. The constants for PO oxidation were higher in comparison to PT. pH dependence revealed two or three ionizable groups with pK _a values of 4.9–5.7 and 7.7–9.7 that significantly affected the activity of peroxidase. Single-turnover experiments showed that the limiting step of PT oxidation was reduction of compound II and second-order rate constants were obtained which were consistent with the constants at steady-state conditions. Laccase-catalyzed PT and PO oxidation rates were lower; apparent bimolecular rate constants varied from 1.8×10⁵M⁻¹s⁻¹ to 2.0×10⁷M⁻¹s⁻¹ at pH 5.3. PO constants were higher in comparison to PT, as was the case with peroxidase. The dependence of the apparent bimolecular constants of compound II or copper type 1 reduction, in the case of peroxidase or laccase, respectively, was analyzed in the framework of the Marcus outer-sphere electron-transfer theory. Peroxidase-catalyzed reactions with PT, as well as PO, fitted the same hyperbolic dependence with a maximal oxidation rate of 1.6×10⁸M⁻¹s⁻¹ and a reorganization energy of 0.30 eV. The respective parameters for laccase were 5.0×10⁷M⁻¹s⁻¹ and 0.29 eV. Received: 20 September 1999 / Accepted: 24 February 2000     

Coiled-coil nanomechanics and uncoiling and unfolding of the superhelix and alpha-helices of myosin

The nanomechanical properties of the coiled-coils of myosin are fundamentally important in understanding muscle assembly and contraction. Force spectra of single molecules of double-headed myosin, single-headed myosin, and coiled-coil tail fragments were acquired with an atomic force microscope and displayed characteristic triphasic force-distance responses to stretch: a rise phase (R) and a plateau phase (P) and an exponential phase (E). The R and P phases arise mainly from the stretching of the coiled-coils, with the hinge region being the main contributor to the rise phase at low force. Only the E phase was analyzable by the worm-like chain model of polymer elasticity. Restrained molecular mechanics simulations on an existing x-ray structure of scallop S2 yielded force spectra with either two or three phases, depending on the mode of stretch. It revealed that coiled-coil chains separate completely near the end of the P phase and the stretching of the unfolded chains gives rise to the E phase. Extensive conformational searching yielded a P phase force near 40 pN that agreed well with the experimental value. We suggest that the flexible and elastic S2 region, particularly the hinge region, may undergo force-induced unfolding and extend reversibly during actomyosin powerstroke.     

Synthesis of hapten and conjugates of coumestrol and development of immunoassay

3-O-Carboxymethylcoumestrol was prepared as the hapten for immunoassay by a partial alkylation of coumestrol with ethyl chloroacetate in acetone alkalized with potassium carbonate. 3-O-Ethoxycarbonylmethylcoumestrol was separated by column chromatography and finally was hydrolyzed with formic acid. 1H and 13C NMR data (APT, COSY, HMQC, and HMBC) revealed that the reaction was regioselective, as 3-O-ethoxycarboxymethylcoumestrol was the only monosubstituted derivative. The hapten was then conjugated to bovine serum albumin and used for immunization of rabbits. A radioimmunoassay (RIA) system was established based on the polyclonal antiserum and a 125I-labeled hapten-tyrosine methyl ester conjugate as the radioligand. Parameters of the RIA: sensitivity: 12 pg per tube, 50% intercept: 140 pg per tube, working range: 20-4000 pg per tube. The cross-reactivity of a panel isoflavonoid and lignan phytoestrogens was either negligible (e.g. formononetin 0.07%; biochanin A 0.06%) or not detectable at all. The major immunoreactive peak in HPLC fractions from an alfalfa extract had the same retention time as coumestrol standard and represented 94.8% of the signal. The remaining 5.2% of immunoreactivity was distributed between five minor peaks. We conclude that after the validation for particular matrices, the method will be a useful tool for analysis of coumestrol, especially in low volume and low concentration samples.     

白术同源四倍体的诱导和鉴定及其与二倍体过氧化物酶的比较

以白术（Atractylodes macrooephala Koidz.）二倍体组培苗为材料,对其四倍体诱导方法进行研究,共获得45个白术同源四倍体株系,为优良株系的选育提供了材料。此外,还分析比较了其中8个白术四倍体株系与二倍体的过氧化物酶同工酶（POD）的酶谱差异,发现四倍体各株系过氧化物酶同工酶谱比二倍体的均多了Rf0.310的谱带,且总过氧化物酶比活力也发生了很大改变,对探讨白术四倍体优良株系的生理生化机理具有一定的参考价值。     

Effect of soil salinity and water stresses on growth and content of nitrogen,chloride and phosphate of wheat and triticale

Summary Three wheats and one triticale were grown, up to flowering stage, in pots on calcareous soil adjusted to a range of salinities (S₁=3.5, S₂=6, S₃=8.5, and S₄=11 mmhos/cm, 20°C, soilpaste extract) by adding solution consisting of 3∶2∶1 of Na-, Ca- and Mg chlorides in chemical equivalent amounts. Moisture in the pots was kept at 100% (W₁), 40% (W₂) and 20% (W₃) of the available water. The vegetative growth, nitrogen and phosphate were affected by S and W treatments, chloride was affected only by S. The interaction S×W affected only dry weight. Varietal effect was observed between wheat as a group and triticale. Multiple quadratic regression equations of these properties on salinity and water revealed that the higher the available water the wider the range of tolerable salinity. Triticale was relatively more tolerant to water stress. Salinity increases Cl and decreases N, whereas water stress enhances N accumulation to a certain extent. However, in triticale at S₃ and S₄ the effect of water stress on N was overshadowed by the excessive salinity. This did not occur for the wheat (Florence). P trends were described. R² for P was low (0.7435–0.3603) which made interpretations rather difficult.     

放牧对牧草光合作用、呼吸作用和氮、碳吸收与转运的影响

研究放牧对草地植物生理活动的影响,对于揭示草地放牧演替的生理机制有重要意义.大量研究表明,家畜放牧对牧草光合作用、呼吸作用以及C和N吸收与转运的影响,可以分为生理伤害和生理恢复2个阶段.放牧通过改变草地冠层结构影响牧草光合作用,净光合作用速率短期内迅速下降,随着叶面积指数增加又逐渐上升,呼吸作用有相似的变化趋势.牧草放牧后再生长所需的C和N最初主要来自根系和留茬中的贮藏物质,此后随着牧草生长恢复逐渐由同化作用供给,C代谢与土壤N水平负相关.放牧后牧草生理活动变化与牧草遗传特性、种间竞争、家畜放牧特征、非生物环境等因素密切相关.