Isotopic fractionation during N2 fixation by four tropical legumes

To quantify the contribution of biological nitrogen fixation (BNF) to legume crops using the ¹⁵N natural abundance technique, it is necessary to determine the ¹⁵N abundance of the N derived from BNF—the B value. In this study, we used a technique to determine B whereby both legume and non-N₂-fixing reference plants were grown under the same conditions in two similar soils, one artificially labelled with ¹⁵N, and the other not. The proportion of N derived from BNF (%Ndfa) was determined from the plants grown in the ¹⁵N-labelled soil and it was assumed that the %Ndfa values of the legumes grown in the two soils were the same, hence the B value of the legumes could be calculated. The legumes used were velvet bean (Mucuna pruriens), sunnhemp (Crotalaria juncea), groundnut (Arachis hypogaea) and soybean (Glycine max) inoculated, or not, with different strains of rhizobium. The values of %Ndfa were all over 89%, and all the legumes grown in unlabelled soil showed negative δ¹⁵N values even though the plant-available N in this soil was found to be approximately +6.0‰. The B values for the shoot tissue (B_s) were calculated and ranged from approximately −1.4‰ for inoculated sunnhemp and groundnut to −2.4 and −4.5‰ for soybean inoculated with Bradyrhizobium japonicum strain CPAC 7 and Bradyrhizobium elkanii strain 29W, respectively. The B (B_wp) values for the whole plants including roots, nodules and the original seed N were still significantly different between the soybean plants inoculated with CPAC 7 (−1.33‰) and 29W (−2.25‰). In a parallel experiment conducted in monoxenic culture using the same soybean variety and Bradyrhizobium strains, the plants accumulated less N from BNF and the values were less negative, but still significantly different for soybean inoculated with the two different Bradyrhizobium strains. The results suggest that the technique utilized in this study to determine B with legume plants grown in soil in the open air, yields B values that are more appropriate for use under field conditions.     

植物病原真菌尖孢镰刀菌检测与定量研究进展

尖孢镰刀菌（Fusarium oxysporum）是一种危害严重的土传病原真菌,被列为世界十大植物病原真菌之一。该菌能够侵染棉花（Gossypium hirsutum）、大豆（Glycine max）、西瓜（Citrullus lanatus）、香蕉（Musa nana）、番茄（Lycopersicon esculentum）和苜蓿（Medicago sativa）等100多种具有重要经济价值的作物,引起枯萎病和根腐病等。对土壤和植物根组织中的尖孢镰刀菌进行检测和定量是病害早期诊断和有效防治的基础。本文对国内外关于尖孢镰刀菌检测和定量方法（主要包括培养基菌落平板稀释法、常规PCR和实时荧光定量PCR等）及其应用进行总结,旨在对农牧业生产中尖孢镰刀菌检测和定量提供理论指导。     

辐射松根朽病与林龄和环境条件关系的数量化分析

对新西兰辐射松根朽病（Ａｒｍｉｌｌａｒｉａｓｐｐ．）的发病程度与林令和环境条件的关系进行了数量化分析。结果表明影响该病发生的主要因素有：年降雨量、地形、土壤类型、前作以及它们之间的互作关系。以此建立的多无线性回归模型为：＝－４．３＋５．４Ｒ２＋１２．４ｓ４＋７．８ｐ１＋６．７ｐ２＋２９Ｌ２＋３１．１Ｒ２ｐ１Ｌ２．根据这一模型可预测新西兰境内任一给定的条件下根朽病的可能侵染水平,以供造林设计和防治时参考。     

Factors associated with variation in Neospora caninum bulk-milk S/P ratios in initially bulk-milk negative testing Dutch dairy herds

We conducted a study on 81 initially bulk-milk ELISA negative dairy herds taken from a random sample of Dutch dairy herds to evaluate variation in bulk-milk S/P ratios and to study reasons for bulk-milk conversion. These herds were repeatedly (3-month intervals) tested between April 2004 and August 2005 and serostatus of all animals had previously been established as negative (N), low-positive (LP) or high-positive (HP). Of these herds, herd- and test-related factors associated with variation in sample over positive (S/P) ratios were analysed using a multivariable linear-mixed model with ‘herd’ as random effect. In addition, changes of animal serostatus in converting herds were described. S/P ratios were calculated as the optical density of the bulk-milk sample minus the optical density of the negative serum control divided by the difference in optical density between the positive and negative serum control.

Sixteen bulk-milk conversions in 12 dairy herds were seen with few indications of serological conversion in lactating cattle except for one herd in which recrudescence of infection appeared likely in nine cows. The effect of HP serostatus on bulk-milk S/P ratio was 2–3 times stronger compared with LP serostatus. In addition, bulk-milk S/P-ratio increased when the proportion of HP animals between 1 and 60 days in milk increased and decreased when the average milk-production level of the herd increased. Besides these herd-related factors, the use of different ELISA-testkits between test rounds had a significant effect on the S/P-ratio in bulk-milk samples.     

Tracking fungi in soil with monoclonal antibodies

Species of the genus Trichoderma are ubiquitous soil-borne fungi that exhibit antagonism towards a number of economically important plant-pathogenic fungi and oomycetes. This review discusses recent developments in the use of monoclonal antibodies to detect these fungi in their natural soil environments and to quantify their population dynamics during antagonistic interactions with saprotrophic competitors in soil-based systems. Immunological approaches to detection and quantification are examined in relation to conventional plate enrichment techniques and to nucleic acid-based procedures. An example of recent research using a mAb-based assay to quantify the effects of saprotrophic competition on the growth of Trichoderma isolates in mixed species, soil-based, microcosms is presented. Future technological developments in immunoassays for tracking Trichoderma populations in soil are discussed and results presented showing the accurate detection and visualization of a plant growth-promoting isolate of T. hamatum in the rhizosphere of lettuce using mAb-based immunodiagnostic assays.     

Quantification of DNA of citrus huanglongbing pathogen in diseased leaves using competitive PCR

A quantification system for huanglongbing pathogen using a competitive polymerase chain reaction method and image-analyzing software were developed to obtain precise results. Significant differences in the quantity of pathogen were thus determined in leaves of two citrus cultivars commonly cultivated in southern Vietnam. Less pathogen-related DNA was detected from the tissue of citrus cultivars that are believed to be more tolerant than susceptible cultivars. The quantification system will be used in studies on pathogen proliferation and movement inside citrus tissue.     

天然次生林群落垂直结构的定量研究

本文以三个次生林群落类型为对象,运用相对多度(RA)、相对频度(RF)、相对显著度(RD)及重要值(Ⅳ)等指标,对其垂直结构进行了深入分析,运用填充系数(F)对群落的层次结构研究提供重要依据;在参考多种分层系统的基础上运用动态的分层方法对群落的层次结构进行了较为合理的划分;分析了次生林群落的垂直结构特征对于群落生产力及其动态演替趋势的意义.     

土地利用对水体营养物影响的研究进展

日益严重的水质污染威胁到人类健康与生存,水体富营养化是其中一大治理难题。流域土地利用深刻影响水体中的营养物,定量刻画两者关系,可有效指导土地利用优化以改善水体质量。在国内外研究的基础上,本文梳理了样地实验、统计分析、经验模型和机理模型等4类研究土地利用和水体营养物关系的方法,并分析了不同方法应用的优缺点。以往研究多侧重土地利用数量结构,对强度和空间分布的量化及其与水体营养物关系的模型构建等研究尚显不足,多从单一或部分信息进行松散研究,导致不同研究中土地利用对水体营养物的解释能力差异显著。本文着重综述土地利用类型、强度差异和空间分布等3个方面对水体营养物影响的研究进展,并阐述关于两者定量关系尺度效应的争议和不确定性。目前主要存在问题包括现有研究土地利用和水体营养物的关系手段不足、土地利用多组分信息和水体营养物关系规律不清和尺度效应不确定性等3方面。因此,本文以水体污染物的产生、迁移、转化等的生态水文过程为线索,提出土地利用多信息空间综合表达和量化研究框架作为核心突破口。应用该框架,未来围绕融合过程机理与经验统计的模型开发、富营养化过程关联的土地利用多信息综合量化和多尺度结构的构建等3个方面开展研究,旨在为丰富土地利用对水体营养物影响研究提供一定的参考价值。     

Quantitative analysis of biochar in field soil

Biochar is used with increasing frequency as a soil amendment because of its potentially beneficial effects on soil carbon sequestration, crop yield, nutrient leaching and greenhouse gas emissions. Simple methods for the analysis of biochar in soil, however, are currently unavailable. Therefore, we have adapted the “loss on ignition” method for this purpose. The technique requires knowledge of the proportions of both biochar and biochar-free soil that are lost on ignition. One can use values determined prior to the amendment of the soil with biochar, assuming that the values do not change after biochar is incorporated in the soil. We tested these assumptions. Over the course of 15 months, the assumptions proved to be valid under our test conditions. The technique accurately determined a wide range of biochar concentrations in field soil.     

虾类行动障碍野田村病毒(MDNV) TaqMan RT-PCR检测方法的建立与应用

基于虾类行动障碍野田村病毒(movement disorder nodavirus, MDNV)的RNA依赖的RNA聚合酶(RdRp)基因设计寡核苷酸引物和探针,以含有MDNV靶基因的pMD18-MDNV质粒和RNA标准品为模板,通过优化反应体系和反应程序,建立了MDNV的TaqMan RT-PCR检测方法。优化后的反应组分：20.0 μL TaqMan RT-PCR反应液预混液中包含11.0 μL一步法RT-PCR缓冲液、0.8 μL酶混合物、0.3 μmol/L正向引物、0.3 μmol/L反向引物、0.4 μmol/L探针、1.0 μL模板和5.2 μL RNA-free H2O;优化后的反应程序：54.5℃ 15 min,95℃ 1 min;45个热循环扩增(95℃ 10 s,60.3℃ 30 s)。本研究所建立的方法能实现对MDNV的特异性检测,在1.4×1010~1.4×101 copies/μL标准质粒浓度范围内,起始模板浓度对数值与反应循环数间呈良好的线性关系;该方法最低可检测5.5×101拷贝的RNA标准品或1.4×101拷贝的pMD18-MDNV标准质粒。分析结果还显示,该检测方法批内Ct值和批间Ct值的变异系数(CV)分别小于1.27%和1.83%,具有良好的重复性和稳定性。利用该方法对2019年采自我国部分省市的样品进行检测发现,凡纳滨对虾(Litopenaeus vannamei)中MDNV的阳性检出率为23.5% (16/68)。本研究建立的MDNV TaqMan RT-PCR方法具有特异性、快速和灵敏等特点,可为对虾养殖实践中MDNV的定性、定量检测与监测以及有效防控提供技术支持。