柑橘木虱对12种寄主植物嫩梢及其挥发物成分的趋性反应

为了发现对柑橘木虱Diaphorina citri Kuwayama具有引诱活性的寄主植物嫩梢挥发物,采用Y型嗅觉仪在室内测定柑橘木虱对12种寄主植物嫩梢及其挥发物成分的趋性反应,采用固相微萃取技术(SPME)和气-质联用分析技术(GC-MS)分析鉴定嫩梢挥发物的共同成分.结果表明,柑橘木虱对九里香Murraya ex...     

柑橘木虱胁迫对柠檬叶代谢组挥发性成分的影响

柑橘木虱是柑橘黄龙病的自然传播媒介,为了解柑橘应对柑橘木虱胁迫的代谢组学变化,本文以柠檬为受体,进行柑橘木虱取食胁迫处理,分别于胁迫不同阶段(0、12、24、48、72和96 h)采集柠檬叶为试验材料,静态顶空-气相色谱-质谱联用检测其挥发性物质。主要结果如下:结合保留指数从柠檬叶中共检出37种化合物,包含15种萜类,6种醛类,6种醇类等;含量较高的挥发性成分有(+)-柠檬烯、桧烯、月桂烯等;受柑橘木虱取食胁迫后,柠檬叶中的挥发性化合物含量产生明显变化,萜类、醛类和醇类物质相对含量均呈升高趋势。桧烯、里那醇、(+)-香茅醛和橙花醇4种物质的含量在柑橘木虱取食胁迫96 h时分别升高2.1,2.9,7.8和3.2倍;α-石竹烯的含量下降1.9倍。(+)-柠檬烯、β-红没药烯和反式-α-香柑油烯等物质含量变化不明显,胁迫后的样品中新检出玫瑰呋喃氧化物、香叶醇和2-异丙基-5-甲基茴香醚等3种化合物。皮尔森相关性分析表明,桧烯、β-蒎烯、甲基庚烯酮、月桂烯、水芹烯等9种物质含量与柑橘木虱胁迫时间呈极显著正相关(P0.01),α-蒎烯、莰烯、松油烯、γ-萜品烯等10种物质含量与柑橘木虱胁迫时间呈显著正相关(P0.05),而邻甲氨基苯甲酸甲酯含量与柑橘木虱胁迫时间呈极显著负相关(P0.01)。聚类和热图进一步分析表明,胁迫不同时间的样品具有显著差异,且胁迫时柠檬叶挥发性化合物大部分呈现上升的趋势,尤其是胁迫48、72和96 h后,物质含量显著增加。研究结果为了解柑橘应对柑橘木虱等生物胁迫的代谢组学变化及柑橘木虱的生物防治提供了理论基础。     

健康和感染黄龙病沙糖桔嫩梢挥发性成分的分析

亚洲柑桔木虱Diaphorina citri Kuwayama为黄龙病的主要媒介昆虫。与健康柑桔相比,带有嫩梢并感染黄龙病的柑桔对该木虱的吸引作用较强。本文采用顶空固相微萃取和气质联用(GC/MS)分析健康和黄龙病沙糖桔嫩梢挥发物成分。结果表明,健康沙糖桔和黄龙病沙糖桔挥发性主要成分都是烯烃类、醇类,其中健康柑桔嫩梢挥发物的主要组分为β-榄香烯(17.0%)、β-罗勒烯(15.22%)、β-水芹烯(14.29%),而黄龙病嫩梢挥发物的主要组分为β-水芹烯(59.98%)、β-榄香烯(9.04%)。健康和黄龙病沙糖桔释放的挥发性化学物质种类差异较大,同种化合物的含量差异也较大。     

受害马尾松、湿地松挥发性化学物质的释放

萜烯类化合物是针叶树防御昆虫为害的重要组分。对盆栽马尾松、湿地松进行未受害、接虫咬食为害、人工剪叶处理后,用TCT-GC-M S分析了叶片挥发物的成分及相对含量。结果表明:人工剪叶后马尾松挥发物相对含量除α-蒎烯降低外,其余都有所升高,但挥发性化学物质相对含量出现高峰的时间没有规律性;虫害后,挥发物的相对含量除水芹烯降低外,其余均在1h升高,同时检测到p-薄荷-1.4(δ)-二烯,但其变化较小。湿地松受害后检测到7种挥发性化学物质,除人工剪叶后α-蒎烯、虫害后α-蒎烯、水芹烯相对含量降低外,其它挥发物的相对含量在不同的时间内都有所升高。β-蒎烯在两种松树受害后都有显著的变化,马尾松中的石竹烯没有明显的变化,氨茴酸-3,5-二甲基-3-乙烯基-4-己烯酯、大香叶烯D只在湿地松中检测到,而且其变化显著。     

钝叶榕榕果挥发物成分及其构成特征分析

为揭示钝叶榕与其传粉榕小蜂及两种协同进果的非传粉榕小蜂Diaziella yangi和Lipothymus sp.间的化学联系与分配机制,采用顶空动态法提取钝叶榕(Ficus curtipes Corner)雌花期和传粉后的榕果挥发物,用定性和定量的方法鉴定、分析了挥发物的成分和变化动态。结果表明:从钝叶榕榕果中共鉴定出45种挥发物成分,主要是单萜类和倍半萜类化合物。6-甲基-5-庚烯-2-酮、β-罗勒烯、反-β-金合欢烯、α-金合欢烯、α-派烯、香桧烯、顺-β-罗勒烯、顺-β-香柠檬烯、大香叶烯D和4,8-二甲基-1,3,7-壬三烯可能是构建钝叶榕特异性化学信息的基础。雌花期榕果挥发物释放量明显高于传粉后挥发物的释放量,且两者在时间和空间上存在异质性。这说明榕蜂育幼繁殖系统间存在着化学信息的联系。     

稻虱缨小蜂对水稻品种挥发物的行为反应

四臂嗅觉仪的行为试验表明、稻虱缨小蜂（Anagrus nilapareatae Pang et Wang)对不同水稻品种挥发物的行为反应存在显差异。在测试的6个未受害水稻品种中。浙852和TN1对稻虱缨小蜂的引诱作用显强于Nabeshi。受褐飞虱[Nitapareata lugens(Stal)]为害后．品种间引诱作用的差异更趋明显。在同一品种内,稻虱缨小蜂对受橱飞虱为害后的稻椿挥发物比未受害的稻株挥发物具更强的行为反应。然而．在稻椿、褐飞虱若虫复合体与稻株、褐飞虱卵,雌成虫复合体的挥发物间不存在显差异。     

假臭草花精油的化学组成及对柑橘木虱的驱避和致死活性

【目的】为了让入侵植物假臭草Praxelis clematidea(Griseb)R.M.KingH.Rob.变废为宝,首次探讨了假臭草花精油对柑橘木虱Diaphorina citri Kuwayama的驱避和致死活性,以用于生物防治。【方法】采用水蒸气蒸馏法提取了假臭草不同部位的精油,并通过气相色谱-质谱联用仪(GC-MS)分析花精油的化学成分,并检测了其对柑橘木虱的生物活性。【结果】假臭草花精油得率(0.31±0.03)%高于茎叶精油的得率(0.23±0.02)%;GC-MS检测显示,假臭草花精油至少含有25种化合物,其中β-荜澄茄油烯(43.85%)和β-石竹烯(30.34%)是其主要成分;花精油浓度为40和50μL·mL~(-1)处理30 min时,对柑橘木虱的驱避反应率均可达到100%。不同浓度的花精油对柑橘木虱具有明显的致死活性。当浓度为50μL·mL~(-1)的时候,浸液法处理九里香Murraya exotica L.嫩梢18 h时,柑橘木虱校正死亡率分别达到(56.25±4.79)%,LC50值为44.83μL·mL~(-1),而熏蒸法48 h时的柑橘木虱校正死亡率仅为(66.33±5.00)%,LC50为24.80μL·mL~(-1)。假臭草花精油对柑橘木虱的驱避和致死活性可能与其主要成分的作用有关。【结论】研究结果证明假臭草花精油对柑橘木虱具有良好的驱避和致死活性,具有开发成为柑橘木虱驱避剂及杀虫剂的良好潜能。     

不同种类寄主对柑橘黄龙病菌侵染能力及虫媒适应性的影响

柑橘木虱Diaphorina citri是柑橘黄龙病菌“Candidatus Liberibacter asiaticus”的主要媒介昆虫,探明不同植物种类寄主对柑橘黄龙病菌侵染能力及柑橘木虱适应性的影响,在设计黄龙病防控策略及寄主抗性研究中具有参考价值。本文选取蕉柑Citrus reticulata、蜜柚Citrus grandis、黄皮Clausena lansium和九里香Murraya exotica为研究材料,对比其田间植株黄龙病发病率和柑橘木虱数量及带菌情况,并在实验室条件下比较柑橘木虱对不同寄主的适应性（包括发育历期、存活率、携菌能力和传菌能力）。结果发现：（1）田间蕉柑和蜜柚的感染率最高,分别为47%和37%;而黄皮和九里香的带菌率较低,分别为18%和4%,且菌浓度均较轻微;（2）田间九里香上柑橘木虱卵、若虫及成虫数量均最多,蕉柑和黄皮其次,蜜柚最少,但蕉柑和蜜柚上柑橘木虱的携菌率和携菌量最高;（3）4种寄主上柑橘木虱卵和1~3龄若虫的发育历期没有显著性差异,柑橘木虱4龄、5龄若虫和总若虫历期在九里香上最短,而在蜜柚上最长;4种寄主上4龄、5龄若虫存活率没有显著性差异,但蜜柚上木虱卵的孵化率、1~3龄及总若虫存活率最低。（4）蕉柑和蜜柚上带菌木虱的携菌能力及传菌能力最高,其次为黄皮,九里香最低。该研究证明了除了寄主植物本身抗病性差异外,虫媒对寄主植物的适应性差异也是造成田间黄龙病发病差异的重要因素。     

二球悬铃木叶片挥发物与悬铃木方翅网蝽的相互作用研究

[目的]为了理解悬铃木方翅网蝽Corythucha ciliata(Say)与二球悬铃木间的和昆虫与寄主互作机制,本实验测定了悬铃木方翅网蝽取食对其寄主植物二球悬铃木的挥发性化合物的影响,以及该虫对这些化合物的行为反应。[方法]运用固相微萃取结合气相色谱与质谱联用技术,测定了不同为害时间段中悬铃木叶片挥发物的变化。通过"Y"形嗅觉仪检测了该虫对不同处理的悬铃木叶片和其挥发物成分的行为反应。[结果]随着为害时间的增加,挥发性化合物的种类和数量总体呈上升趋势。悬铃木未处理叶片中的挥发物有4种成分,包括:顺-3-己烯醇,1,8-桉叶素,(E)-4,8-二甲基-1,3,7-壬三烯和反式-β-石竹烯。在为害后叶片中,共检测到9种挥发性化合物,除了上述4种成分外,新增加了α-侧柏烯,桧烯,月桂烯,顺-3-己烯醇乙酸酯和γ-松油烯。行为学研究发现,悬铃木叶片能显著地吸引悬铃木方翅网蝽,其中顺-3-己烯醇,1,8-桉叶素对该虫具有吸引作用,而反式-β-石竹烯具有趋避作用。[结论]悬铃木叶片被悬铃木方翅网蝽危害后会产生多种挥发物,其中一些挥发物能显著影响该虫的行为,可能在悬铃木方翅网蝽的寄主定位中起着重要的作用,另一些化合物可能参与悬铃木对该虫的间接防御反应。     

不同寄主植物对柑橘木虱发育和繁殖的影响

【目的】研究柑橘木虱Diaphorina citri Kuwayama在柚、酸橘、黄皮、九里香、砂糖橘5种代表性芸香科寄主植物上的发育、存活和繁殖情况,为柑橘木虱及黄龙病的可持续防控提供参考。【方法】利用种群生命表的方法,分析了柑橘木虱在5种不同寄主植物上的发育历期、存活率、成虫寿命、性比及产卵量等数据。【结果】柑橘木虱卵、1龄若虫以及整个若虫的发育历期受寄主植物的影响较为明显,在26℃条件下在柚子植物上柑橘木虱若虫的存活率最高(58.10%),在黄皮上最低(46.04%),两者差异显著。寄主植物影响柑橘木虱成虫寿命,在柚子上柑橘木虱成虫的寿命显著长于黄皮上的寿命。黄皮上的柑橘木虱的单雌产卵量(298粒/雌)显著低于其他4种寄主植物。柑橘木虱在九里香上的内禀增长率(rm)最高(0.133 7)、酸橘上最低(0.129 8);而净增值率(R0)在砂糖橘上最高(187.74)、黄皮上最低(145.27)。【结论】在5种寄主植物中,除黄龙病隐症寄主九里香之外,显症寄主中砂糖橘是柑橘木虱的最适寄主。     

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.