植物化感作用的研究方法及影响因素

介绍了植物化感作用研究的几种生物测定方法,并对影响化感作用效应的因子进行了论证     

生化他感作用研究中的生物测定方法

为了提高生化他感研究的可靠性、可比性等,应不断地发展和完善生物测定技术,建立合理的生物测定技术系统．为此,讨论了生化他感物质的提取和试样的制备、指示植物的选择、合适的评价指标和统计方法的建立等,为生化他感作用研究中建立系统的生物测定方法提供参考．     

地黄对芝麻的化感作用及其化感物质的分离鉴定

通过分离地黄(Rehmannia glutinosa)组织中抑制芝麻(Sesamum indicum)生长发育的化感物质, 以弄清地黄茬种植芝麻造成芝麻严重减产的原因。以芝麻为受体的生物测定为检测跟踪手段, 通过浸提、萃取、硅胶柱层析分离方法及气相色谱-质谱(GC-MS)鉴定方法, 分离鉴定了地黄组织影响芝麻的化感物质。对地黄不同组织的浸提液进行的生物测定结果表明: 影响芝麻生长发育的化感物质在地黄叶中含量高。地黄叶浸提液的不同有机相萃取组分的生物测定结果表明: 乙酸乙酯相化感活性最强, 乙酯乙酯相通过硅胶柱层析分离获得的洗脱组分Fr7化感活性最强。通过GC-MS及验证试验证明: 地黄抑制芝麻生长的化感物质是月桂酸和2,6-二叔丁基苯酚。研究表明地黄他感作用与自毒作用的化感物质是不同的。     

假臭草水浸提液对4种作物的化感作用

【背景】假臭草是近几年来入侵我国南方的外来杂草,目前已成功侵占海南、福建、广东等省沿海的大部分果园、草地、农田等农业生态系统,给当地农业生产造成极大的危害。【方法】采用室内生物测定方法研究不同浓度的假臭草水浸提液(0.010、0.025、0.050 g·m L~(-1))对4种作物(西瓜、水稻、萝卜和辣椒)种子萌发和幼苗生长的化感作用。【结果】各浓度的假臭草水浸提液对4种作物种子萌发和幼苗生长都有较强的化感作用,并且随着浸提液浓度的增大,化感效应显著增强。当浓度为0.050 g·m L~(-1)时,4种受体作物的化感综合效应顺序为西瓜萝卜水稻辣椒。【结论与意义】明确假臭草对不同供试作物的化感作用,能够为该入侵杂草的科学防治提供理论依据。     

三七自毒与化感作用初步研究

采用生物测定方法,对三七的自毒与化感作用进行初步研究。结果表明:(1)三七种子萌发过程中的自毒作用随其播种密度不同而有一定差异,但无明显规律性。三七种子萌发过程中的分泌物对油菜生长具有化感作用,主要表现为对油菜种子发芽率、发芽指数及苗高的抑制效应;(2)三七水浸液对不同受体植物的化感作用不尽相同,对小麦主要表现为对苗鲜重、苗干重、根鲜重、苗高及须根数有不同程度的促进或抑制作用;对油菜则表现为对其种子发芽率具有抑制作用,而对苗鲜重、苗干重、根鲜重、最长根长具有促进作用;(3)三七存在明显的化感自毒作用,其自毒物质可能为影响其连作的一个重要因素。     

高寒草场优势杂草黄帚橐吾水浸液对牧草的化感作用

研究了高寒草场中主要毒杂草——黄帚橐吾水浸液对同域分布的5种牧草的化感作用.将反应指数(RI)相加平均后分为一、二、三级敏感指数,分别评价不同测定指标、不同发育期和物种水平对化感作用的敏感性;从供体角度对相应RI值做同样处理,以综合评价不同水浸液及其不同浓度的化感效应.结果表明,5种牧草对黄帚橐吾化感作用的敏感性从强到弱依次为早熟禾>大雀麦>中华羊茅>羊茅>垂穗披碱草;牧草种子萌发期的敏感性强于幼苗生长期,尤以萌发指数受化感作用的影响明显.黄帚橐吾根、叶水浸液对牧草生物测定结果大多达到显著差异水平,物种水平的化感效应均表现为抑制作用,说明雨水淋溶是黄帚橐吾向环境中释放化感物质的途径之一.黄帚橐吾根部化感效应强于叶片,可能与其地下种间激烈的资源竞争有关.化感作用在增强黄帚橐吾生存竞争力、扩大种群和入侵草场中起着不容忽视的作用,可能是黄帚橐吾单优势种群落形成和草场退化的原因之一.     

培养基组分对沙打旺(Astragalus adsurgens Pall)组培根增殖的影响及其培养滤液提取物的化感活性

采用L9 (315 )正交设计,研究了B5培养基营养组分对沙打旺组培根增殖的影响;并采用玻璃皿滤纸培养法,对其培养滤液提取物进行生物测定以验证沙打旺组培根的化感活性.结果显示:培养基的所有营养组分中,Fe2 对沙打旺组培根增殖的影响最大,蔗糖、H2PO 4、 Mg2 、 Mn2 、 Cu2 、 Zn2 、 BO3-3、 Co2 、 I-、C8H12ClNO3 C12H18Cl2N4OS C6H5O2N C6H12O6的影响次之,氮、Ca2 、MoO2-4 和NAA的影响最小.根据不同养分条件下沙打旺组培根干重的极差分析,筛选出适宜沙打旺组培根快速增殖的优化培养基.培养滤液提取物的生物测定结果表明沙打旺组培根培养过程中可能产生化感物质;化感作用强度的差异预示营养胁迫可能影响其化感物质的产生.研究为沙打旺组培根再生与繁殖提供一定依据,并揭示养分条件可能是该植物表达化感作用的影响因素.     

伴生杂草播娘蒿对小麦的化感效应

用室内生物测定方法,对播娘蒿浸提液处理过的不同品系小麦的萌发率、发芽指数、活力指数、根长、苗高、干重、鲜重以及综合效应进行了统计,对作用效果较明显的5个品系进行了丙二醛含量、叶绿素a和叶绿素b比值、胡萝卜素含量的测定以及有丝分裂指数的统计.结果表明,播娘蒿浸提液对所有供试小麦均具有一定的化感效应,但化感效应的强度在不同品系的小麦中存在差异.对效果较明显的5种供试小麦生理指标的测定结果表明,播娘蒿中化感物质的作用位点可能是小麦细胞膜.此外,试验结果还表明,播娘蒿浸提液对小麦的有丝分裂有抑制作用.     

浮游植物的化感作用

生物化感作用研究是近年来兴起的交叉学科,是化学生态学研究的重要领域。研究水域浮游植物化感作用对了解浮游植物之间、浮游植物与其他生物之间的相互作用及作用机理具有重要意义,对了解赤潮和水华的发生机制及其生态控制等具有非常重要的作用。综述了海洋和湖泊浮游植物化感作用和化感物质的内涵,讨论了水体浮游植物化感作用的特点、研究化感作用的基本方法、化感物质的种类以及影响化感物质作用的生物和非生物因素,详细介绍了浮游植物化感物质的作用机理以及逃避和拈抗化感作用的方式,同时对目前研究的热点问题及未来研究的方向做了简要概述。     

喜旱莲子草组织水浸液对黑麦草种子和幼苗的化感效应

以喜旱莲子草(Alternanthera philoxeroides)地上和地下部组织为供体,黑麦草(Lolium perenne L.)为受体,运用室内培养皿生物测定方法研究了喜旱莲子草组织水浸液的化感作用.结果表明:喜旱莲子草组织水浸提液对黑麦草种子的萌发具有明显抑制作用,显著降低了其终萌发率,也明显抑制了黑麦草幼苗根和茎的生长;喜旱莲子草组织水浸提液处理使黑麦草幼苗叶片的电导率显著增加,但其叶绿素含量却明显降低;喜旱莲子草不同组织部位水浸提液的化感作用强弱不同,表现为地下部分水浸提液化感作用明显强于地上部分.     

An improved cage system for the bioassay of Metarhizium anisopliae on Thrips tabaci (Thysanoptera: Thripidae)

The ability of thrips and other minute insects to escape from conventional assay cups led to development of a successful assay chamber. In the present study, we evaluated three bioassay systems for thrips; Tashiro cage, microtube assay and leaf sandwich assay and introduced an improved thrips entomopathogenic bioassay system (TEBS). Our objective was to investigate the effect of Metarhizium anisopliae on Thrips tabaci using an easy-to-use and inexpensive holding chamber that would produce bioassay results with acceptable levels of accuracy and precision. In our assays, we found that escape of second-instar larvae from TEBS was significantly less than those of other methods which we tested. Our results indicated that our assay system was more suitable than the other conventional assays for the bioassay of entomopathogenic fungi on T. tabaci.     

八种杀虫剂对愈纹萤叶甲成虫的毒力及田间防控效果

【目的】为探索适用于愈纹萤叶甲Galeruca reichardti成虫的防治药剂室内毒力测定方法,并筛选其防治的有效药剂。【方法】分别采用浸虫法、浸叶法和胃毒触杀联合毒力法测定了8种药剂对其成虫的毒力,并通过大田试验进一步验证。【结果】浸虫法测得除虫菊素对愈纹萤叶甲成虫的毒力最高,其LC50值为68.24 mg/L;浸叶法测得氟啶虫胺腈的毒力最高,其LC50值为111.11 mg/L;胃毒触杀联合法测得氟啶虫胺腈的毒力最高,其LC50值为67.94 mg/L。其中,除虫菊素毒力值变化最大,浸虫法毒力是浸叶法的28.97倍,胃毒联合法毒力是浸叶法的17.83倍。蛇床子素、苦参碱在3种生测方法中毒力较低。田间试验结果显示,50%氟啶虫胺腈水分散粒剂562.5 g/hm2防治效果达94.90%,对韭菜苗保苗效果为81.45%,且显著高于其他药剂。【结论】3种毒力测定方法均可用于该虫的毒力测定,而从此虫的为害特点来看,胃毒触杀联合毒力法更能显示药剂的毒力性质。在愈纹萤叶甲成虫盛发期选择氟啶虫胺腈喷施可以有效减轻该虫为害。     

水稻化感生物测试方法的比较及应用

选用8个化感作用潜力各异的水稻品种(系)：PI312777、Lemont、Moroberekan、IAC25、IACA7、IAC120、Batatais、Iguape Cateto为供试材料,以田间伴生稗草为受体材料,比较分析了目前较为通用的3种生物测试方法：琼脂迟播共培法(RSA)、石英砂迟播共培法(RSS)和根系分泌物培养法(SRE)对水稻化感潜力的测试效果．结果表明,RSA法应用于测试水稻化感作用潜力的效果最好,RSS法和SRE法的测试结果虽较为一致,但两者都存在不足．据此,运用RSA法对57个水稻材料进行了化感抗草种质资源的初步筛选,从中获得5个强化感作用潜力的水稻材料：Iguape Cateto、PI312777、Azucena、Taichung Native 1和IAC25．     

An appraisal of bioassay methods for the detection of mycotoxins—a review

The literature on bioassay methods for mycotoxin detection has been reviewed. An outline of the range of bioassay methods is given and the role of cytotoxicity tests in particular has been emphasized.     

Assessing the use of known mutagens to calibrate the Salmonella typhimurium mutagenicity assay: I. Without exogenous activation

There has been an increasing need in genetic toxicology to progress from strictly qualitative tests to more quantitative tests. This, in turn, has increased the need to develop better quality assurance and comparative bioassay methods. In this paper, two laboratories tested 10 Salmonella mutagens in order to determine the usefulness of selected chemicals as potential reference materials to calibrate the Salmonella assay. If variance within a bioassay is sufficiently low and the rankings of the compounds are of acceptable consistency, the chemicals later could be evaluated for use as standard control compounds, as audit materials, and as standard reference materials for comparative bioassay efforts. The results demonstrated that the chosen chemicals (with the possible exception of dimethylcarbamylchloride) provide such consistent results in the Salmonella mutagenicity bioassay that they can be used for semi-quantitative calibration and as possible bioassay controls, special audit chemicals, and potentially as reference standards in comparative bioassay efforts. Reference standards, whether used as audit materials or in comparative bioassays, must be used concurrently with the test substances of interest; used without bias; used in a standardized, highly controlled bioassay; and be tested across an appropriate dose range. The study also shows that when these compounds are used as reference standards much care must be given to the number and spacing of doses if highly reproducible slope values are to be generated. We recommend use of a pilot test to establish a dose range for definitive tests and the placement of doses for the definitive tests within the first half of the linear dose-response curve. For appropriate comparisons, one should replicate the tests using the defined dose range and analyze the results in a non-biased statistical manner.     

Testing for bacterial resistance to arsenic in monitoring well water by the direct viable counting method.

Direct viable counting of metal-resistant bacteria (DVCMR) has been found to be useful in both enumerating and differentiating metal-resistant and metal-sensitive strains of bacteria. The DVCMR bioassay was used to detect effects of low and high concentrations of arsenic and arsenicals on bacterial populations in groundwater. The level of resistance of the bacterial populations to arsenate was determined by the DVCMR bioassay, and the results showed a linear correlation with the total arsenic concentrations in the monitoring well water samples; no correlation was observed by culture methods with the methods employed. Bacteria resistant to 2,000 micrograms of arsenate per ml were isolated from all monitoring well water samples studied. Strains showed similar antibiotic and heavy-metal profiles, suggesting that the arsenic was not a highly selective pressure for arsenic alone. The monitoring well water samples were amended with arsenate and nutrients to determine the biotransformation mechanisms involved. Preliminary results suggest that bacteria indigenous to the monitoring well water samples did not directly transform, i.e., precipitate or volatilize, dissolved arsenic. It was concluded that arsenic contamination of the groundwater can be monitored by the DVCMR bioassay.     

How many high production chemicals are rodent carcinogens? Why should we care? What do we need to do about it?

High production volume (HPV) chemicals are produced in or imported to the US in amounts greater than 1 million pounds per chemical per year. The EPA has identified thousands of HPVs. Due to their abundance, such chemicals bring a substantial risk for human exposure, and as a result some level of adverse consequences to health are to be expected. In order to examine the potential for cancer risk associated with HPVs, this paper examines HPVs that have been tested in the National Toxicology Program's rodent cancer bioassay. The chemicals tested in the bioassay represent a small sample of the universe of environmental chemicals to which people are exposed. Unexpectedly, 60% of the 128 HPVs evaluated in the bioassay proved to be rodent carcinogens. This value compares to a predicted prevalence of only 16.5% carcinogens in a previous study. The previous study concluded that HPVs were less likely to be toxic by a variety of other test criteria as well. However, the approach involved identifying putative carcinogens using quantitative chemical structure-activity relationships (QSAR) in contrast to the direct tabulation of bioassay test results performed here. Detailed examination of bioassay results reveals that test outcomes depend heavily on route of administration as well as on dose level, sex, strain, and species used. Since most of these factors have little or no apparent relationship to chemical structure, results of this study suggest that QSAR, as well as virtually all other alternative methods, may not generally provide accurate predictions of carcinogenic potential. As we wait for efficient and effective methods for predicting carcinogens to be developed, people continue to be exposed to environmental carcinogens. Progress on regulating environmental carcinogens as well as on developing more effective test methods has been minimal since "war on cancer" began approximately 30 years ago. The present study questions whether the current strategy for dealing with environmental carcinogens will ever be successful. Close examination of rodent cancer test results seems to suggest that almost all chemicals may have carcinogenic potential in some genotypes under some exposure circumstances. If this hypothesis is correct, it would explain the general lack of progress in developing methods to differentiate carcinogens from noncarcinogens. A completely new strategy for dealing with cancer caused by exposures to environmental chemicals seems to be needed.     

Testing for bacterial resistance to arsenic in monitoring well water by the direct viable counting method

Direct viable counting of metal-resistant bacteria (DVCMR) has been found to be useful in both enumerating and differentiating metal-resistant and metal-sensitive strains of bacteria. The DVCMR bioassay was used to detect effects of low and high concentrations of arsenic and arsenicals on bacterial populations in groundwater. The level of resistance of the bacterial populations to arsenate was determined by the DVCMR bioassay, and the results showed a linear correlation with the total arsenic concentrations in the monitoring well water samples; no correlation was observed by culture methods with the methods employed. Bacteria resistant to 2,000 micrograms of arsenate per ml were isolated from all monitoring well water samples studied. Strains showed similar antibiotic and heavy-metal profiles, suggesting that the arsenic was not a highly selective pressure for arsenic alone. The monitoring well water samples were amended with arsenate and nutrients to determine the biotransformation mechanisms involved. Preliminary results suggest that bacteria indigenous to the monitoring well water samples did not directly transform, i.e., precipitate or volatilize, dissolved arsenic. It was concluded that arsenic contamination of the groundwater can be monitored by the DVCMR bioassay.     

Potential role of ultra-sensitive estradiol assays in estimating the risk of breast cancer and fractures

Radioimmunoassays (RIA) for the measurement of estradiol are sufficiently sensitive to assess the reproductive status of pre-menopausal women but lack sufficient sensitivity for low concentrations found in post-menopausal women. Bioassays have been used in the past to measure low estrogen levels but are impractical for handling high volumes of tests, particularly routine and non-research specimens. In this study, we compared results for estradiol using several different methods including bioassay, RIA, and two tandem mass spectrometry methods. At the lower tertile of estradiol measurements by RIA, the overall excellent correlation with results obtained by tandem mass spectrometry (i.e. r=0.83) was lost (i.e. r=0.29). In addition, results were much lower with bioassay and mass spectrometry than with RIA suggesting that RIA measures undesired noise or estrogen metabolites. The mass spectrometry methods correlate best with isotopic kinetic methods when assessing aromatase inhibition. On this basis, we conclude that mass spectrometry assays are the best option for measurement of low estradiol concentrations. With such assays, greater discrimination should be achievable when using estradiol levels as a predictor of the risks for breast cancer and for fractures.     

Quantification of Endospore Concentrations of Pasteuria penetrans in Tomato Root Material

Six methods for quantification of the endospore concentrations of Pasteuria penetrans from tomato roots are described. Mortar disruption and machine disruption methods gave the highest estimations (endospores per gram of root material) of 83.7 and 79.0 million, respectively. These methods were significantly superior to incubation bioassay (47.7 million), enzymatic disruption (32.1 million), and enzymatic disruption + flotation (25.8 million) methods. A centrifugation bioassay method gave the lowest estimation of 12.7 million.