内蒙古荒漠草原亚洲小车蝗的种群动态

2010—2011年于内蒙古乌兰察布市四子王旗格根塔拉草原对亚洲小车蝗Oedaleus asiaticus(B.-Bienko)种群动态进行了研究。结果表明,亚洲小车蝗种群空间格局为随机分布;亚洲小车蝗于6月中旬开始孵化出土,1～3龄蝗蝻高峰期在6月中下旬至7月初,终见期在7月下旬;4～5龄蝗蝻于6月下旬始见,高峰期在7月上中旬,终见期在7月末;成虫于7月上旬始见,高峰期在7月中旬至8月下旬,终见期在9月上旬。应用最优分割法将亚洲小车蝗种群动态划分为3个阶段:(1)6月中旬,为蝗蝻开始出土期,数量稀少,空间格局为聚集分布或随机分布;(2)6月下旬至7月上中旬,为蝗蝻发生盛期,密度低时为随机分布,密度高时为聚集分布;(3)7月中下旬至9月上旬,为成虫发生期,空间格局为随机分布。     

内蒙古亚洲小车蝗种群遗传多样性的微卫星分析

亚洲小车蝗Oedaleus asiaticus Bei-Bienko是我国北方草原和农牧交错区主要的害虫之一。为了从分子水平评价内蒙古地区亚洲小车蝗种群的遗传多样性和种群间遗传分化, 本文采用8对微卫星引物对内蒙古15个地点的亚洲小车蝗种群进行遗传多样性分析。结果表明： 各位点有效等位基因数为3.4517~13.2881, 多态信息含量值为0.5601~0.8563, Shannon氏多样性指数在0.7018~4.1789之间。15个种群的平均期望杂合度为0.6836, Nei氏期望杂合度为0.5303~0.6513, 群体遗传距离为0.1092~0.4235, 群体分化率Fst平均值为0.1612, 基因流Nm平均值为1.6164。8个微卫星位点均具有较高的多态性, 各种群间的遗传分化水平较大, 基因交流程度中等, 个体间的遗传变异大于种群间的遗传变异。15个不同地点的亚洲小车蝗种群根据遗传距离共聚为6支。种群间遗传分化与地理距离呈正相关关系。高山和沙漠对不同地区亚洲小车蝗种群的迁移具有阻碍作用, 可能是形成遗传分化的主要原因。研究结果从分子水平探索不同地区亚洲小车蝗种群间的内在联系, 为制定亚洲小车蝗的综合治理策略提供了分子生物学的基础资料。     

型变过程中亚洲小车蝗体内保幼激素滴度及其代谢相关基因表达量的变化

【目的】本研究旨在明确亚洲小车蝗Oedaleus asiaticus在不同发育阶段和型变过程中的保幼激素(juvenile hormone, JH) JHⅢ滴度及其代谢相关基因表达量的变化情况，为阐明保幼激素调控亚洲小车蝗型变中的生理功能奠定基础。【方法】采用高效液相色谱法测定散居型和群居型亚洲小车蝗不同发育阶段(4和5龄若虫及1, 4, 7, 10, 13和20日龄成虫)和3龄若虫群居化处理1, 3, 5和7 d时亚洲小车蝗血淋巴中的保幼激素JHⅢ的滴度变化；用qRT-PCR检测在群居化处理亚洲小车蝗3龄若虫1, 3, 5和7 d时保幼激素代谢相关的3种基因保幼激素酯酶(juvenile hormone esterase, JHE)基因、保幼激素甲基转移酶(juvenile hormone methyltransferase, JHAMT)基因和保幼激素环氧水解酶(juvenile hormone epoxide hydrolase, JHEH)基因的表达量；利用JHⅢ浸液处理散居型亚洲小车蝗3龄若虫后进行群居化处理，测定亚洲小车蝗型变率。【结果】JHⅢ在亚洲小车蝗血淋巴中的滴度甚微...     

亚洲小车蝗对内蒙古典型草原3种禾本科植物取食特性研究

通过了解亚洲小车蝗Oedaleus asiaticus的取食特性,为有害生物的防治和草原生态环境的保护提供理论依据。在分析亚洲小车蝗发生与羊草Leymus chinensis、克氏针茅Stipa krylovii及糙隐子草Cleistogenes squarrosa等3种禾本科植物特征参数关系的基础上,通过罩笼试验调查了罩笼内亚洲小车蝗3~5龄蝗蝻的存活率,比较了亚洲小车蝗的取食数量、取食高度及取食率等特性。结果表明,克氏针茅和糙隐子草生态优势度与亚洲小车蝗发生数量显著正相关,亚洲小车蝗喜食克氏针茅和糙隐子草(0.25≤RFN<0.5),少食羊草(0.025≤RFN<0.25)。4龄和5龄蝗蝻对克氏针茅和糙隐子草取食量显著大于羊草。亚洲小车蝗取食高度位于植物中下部(9~10cm),与3种植被高度无关;取食后上部掉落造成损失,特别是对克氏针茅的为害更大;亚洲小车蝗取食克氏针茅和糙隐子草时的存活率显著高于取食羊草时的存活率。结论:以克氏针茅和糙隐子草为优势植物的典型草原更适合亚洲小车蝗的发生与为害,应加强该类草原亚洲小车蝗的监测预警和防控。     

亚洲小车蝗不同地理种群遗传多样性的等位酶分析

采用等位酶电泳技术对亚洲小车蝗8个地理种群遗传多样性和遗传分化进行了研究,并对8种等位酶系统:苹果酸脱氢酶(MDH)、苹果酸酶(ME)、乙醇脱氢酶(ADH)、谷氨酸脱氢酶(GDH)、谷氨酸-草酰乙酸转氨酶(GOT)、异柠檬酸脱氢酶(IDH)、腺苷酸激酶(AK)和己糖激酶(HK)进行聚丙烯酰胺凝胶电泳分析。结果表明:在亚洲小车蝗8个地理种群中共检测到14个基因位点,其中7个位点为多态位点,检测到25个等位基因;种群总体水平多态位点比率P=42.86%,平均有效基因数A=1.786,平均期望杂合度He=0.072,种群平均遗传距离为0.069～0.235。聚类分析表明,遗传距离与地理距离存在一定相关性。亚洲小车蝗8个种群的遗传分化系数Fst=0.086,基因流Nm=3.142,表明亚洲小车蝗种群间有一定程度的遗传分化及基因交流。     

蝗虫微孢子虫病在草原蝗虫优势种种群及空间的分布

在采用蝗虫微孢子虫Nosema locustae防治过的草场中进行抽样调查,研究了草原蝗虫优势种类、混合种群平均密度与蝗虫微孢子虫疾病分布之关系,以及该疾病的空间分布。在防治后的当年,蝗虫微孢子虫疾病的感染率随着混合种群平均密度及靶标蝗虫亚洲小车蝗Oedaleus asiaticus的感病率的下降而降低。但是,次靶标蝗虫如宽须蚁蝗Myrmeleotettixpalpalis(一种中后期发生的种类)其感病率呈上升趋势,表明该疾病可在不同发生期种类蝗虫之间进行有效地传播。病蝗虫在防治后第7d其空间分布呈随机分布(Poisson),第28d 则是聚集分布,第40d时也呈聚集分布。于1993年、1994年对1988年(样区Ⅱ)、1989 年(样区Ⅲ)采用微孢子虫防治过的草场进行抽样调查。结果表明,在二个样区中,二年混合种群平均虫口密度与混合种群的平均感病率呈正相关(相关系数分别为r=0.289, r=0.479)。蝗虫微孢子虫病在主要优势种,如亚洲小车蝗、宽须蚁蝗、白边痂蝗Bryode maluctuosumluctuosum、皱膝蝗Angaracris /I>spp.、毛足棒角蝗Dasyhippus barbipes均有分布。二个样区中的混合蝗虫种群的平均感病率在1994年显著低于1993年。混合蝗虫种群的种类组成也有所变化,与1993年相比,1994年宽须蚁蝗及白边痂蝗的比例上升较大,而亚洲小车蝗的比例下降。经过5—7年的扩散,蝗虫微孢子虫病至少可扩散距防治区1 000m,其扩散方向可能与风及地势等有关。     

内蒙古地区亚洲小车蝗不同地理种群的RAPD分析

采用随机扩增多态性DNA(RAPD)技术对内蒙古地区亚洲小车蝗Oedaleus asiaticus(B.-bienko)9个不同地理种群90个个体进行扩增,8条随机引物扩增共产生了78条带,多态性片段为62条。对Nei′s基因多样性指数和遗传距离进行分析,结果表明:种群间的遗传分化系数为0.2343,即23.43%的遗传变异存在于种群间,种群内的遗传分化系数为0.7657,即76.57%的遗传多样性存在于种群内,群体内遗传多样性大于群体间遗传多样性。用NJ法对这3个种群的Nei′s遗传距离作聚类分析,结果表明亚洲小车蝗不同种群的遗传分化程度与地理距离具有正相关关系。     

黄胫小车蝗受精囊内含物研究

用组织化学、亲和组织化学方法研究了黄胫小车蝗 Oedaleus infernalis Saussure交配前后受精囊内含物的化学组成。结果表明,黄胫小车蝗受精囊内含物有蛋白质、脂类和碳水化合物。交配前后黄胫小车蝗受精囊腔及腺细胞中蛋白质、碳水化合物的含量有较大差异,交配后的含量明显高于交配前的,说明交配活动启动了受精囊腺细胞的分泌,使受精囊腔及受精囊管中积聚大量的碳水化合物及蛋白质。交配前后受精囊脂类含量没有明显变化。用亲和组织化学方法对交配后受精囊进行染色分析,表明受精囊腔内含物的碳水化合物、蛋白质主要以糖蛋白形式存在,糖残基主要有半乳糖、甘露糖及α-葡萄糖。     

意大利蝗Calliptamus italicus(L.)飞行能力的研究

【目的】明确意大利蝗Calliptamus italicus(L.)飞行能力,丰富意大利蝗迁飞生物学,提高预测预报水平。【方法】2014年6—9月间利用昆虫飞行磨测定意大利蝗的飞行能力,研究其不同日龄、卵巢发育等级的飞行能力。【结果】白日(8:00—20:00),意大利蝗雌成虫自2日龄开始飞行,至6日龄(飞行时间18.22 min,飞行距离231.01 m,飞行速度12.59 m·min-1,飞行百分比76.00%)飞行能力达显著最高(P<0.05),6~10日龄飞行能力逐渐降低;雄成虫2日龄的飞行能力显著最高(P<0.05),其飞行时间、飞行距离、飞行速度和飞行百分比分别为0.64 min、13.34 m、15.70 m·min-1、14.29%,2~10日龄的飞行能力逐渐降低。不同等级卵巢与不同日龄的飞行能力的变化规律一致,其中Ⅱ级卵巢(6日龄)的飞行能力显著最高(P<0.05)。【结论】意大利蝗乌鲁木齐地理种成虫具有一定的飞行能力。     

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、...     

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,对于揭未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。     

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.     

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。     

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     

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.     

Synthesis and Antiviral Activity of Ureides and Carbamates of Betulinic Acid and Its Derivatives

Ureides and carbamates of betulinic acid and its derivatives were prepared in good yields by interaction of betulinic acid, betulonic acid, and betulonic acid 3-oxime with amines, amino acids, and alcohols. Ureides of betulonic acid containingL-Val and L-Met residues were found to be effective against herpes simplex type 1 virus.