大丽轮枝菌在茄田土壤中的分布及抽样方法

对茄子黄萎病在田间的空间分布及取样方法进行了研究,并探讨了田间土壤带菌量的准确测定方法。研究结果表明:用分布型指数法测定茄田土壤中微菌核的空间分布型,当菌核数量在8-63 ～39-93 个/g 干土范围内,为聚集分布。当茄田发病率小于75 % 时,病株在田间均属聚集分布;发病率大于75 % 时呈均匀分布。土壤中微菌核的调查方法以棋盘式为最适宜。     

茄田和菜用大豆田烟粉虱成虫数量及空间格局动态差异

运用随机抽样方法进行田间系统调查,并用t检验法、种群聚集强度指标及地统计学方法,对茄田和菜用大豆田烟粉虱成虫数量及空间格局差异进行研究。结果表明,7月7日至8月25日茄田烟粉虱成虫种群增长模型为y=3053e0.0053x(r=0.823*),菜用大豆田烟粉虱成虫种群增长模型为y=2356e-0.1662x(r=-0.9631**),对茄田和菜用大豆田烟粉虱成虫种群数量之间的差异性进行t检验,t值为3.3333,df=7时,t0.05=2.365,两者之间差异显著。这期间两种作物上烟粉虱成虫的田间水平分布和在茄株叶片上的垂直分布均呈聚集分布格局。     

北疆棉花黄萎病发病率调查及土壤中黄萎病菌微菌核数量与种群类型分析

为了解北疆棉花黄萎病的发生情况及发病与土壤中黄萎病菌微菌核数量的关系和病原菌种群类型, 2021年对北疆石河子?奎屯?博乐等8市(县)棉田棉花黄萎病发病率?土壤中黄萎病菌的微菌核数量?菌株种群类型进行了抽样调查?结果表明, 北疆未发生棉花黄萎病的棉田占49.2%, 0%<发病率<5.0%的棉田占32.7%, 发病率≥5.0%的棉田占18%?与2013年?2015年相比, 2021年无病田率分别增加17.7和12.7百分点, 发病率≥5%的棉田分别减少15.7和21.6百分点?从棉田黄萎病发病率与土壤中微菌核数量的关系来看, 整体上北疆棉田棉花黄萎病发病率与微菌核数量相关性不显著(r=0.119 1); 分区域看, 石河子-沙湾片区?奎屯-乌苏片区?精河-博乐片区棉田黄萎病发病率与微菌核数量的相关系数分别为0.033 2?0.007 6?0.062 3, 均无显著相关性; 而呼图壁-玛纳斯片区棉田黄萎病发病率与微菌核数量的相关系数为0.635 7, 呈中度正相关?土壤中的黄萎病菌菌株以菌核型为主, 占57.9%, 菌丝型占23.2%, 中间型占18.9%?用特异性引物进行PCR检测表明, 土壤中黄萎病菌落叶型菌株占97.6%, 占绝对优势?本研究将为北疆棉区棉花黄萎病的综合防控提供理论依据?     

姜瘟田间分布型的研究及其应用

在几年研究姜瘟流行规律的基础上,对姜瘟在田间周年流行时间序列中空间分布型的变化作了比较系统的调查研究。应用7种分析方法进行检验.扩散系数 C 检验结果表明,姜瘟在田间的扩散系数 C 都大于概率为95%的随机分布型 C 值置信区间上限;Taylor 指数法则检验得:S~2=2.4280X~(1·1722);(?)-M 回归方程为:(?)=1.0529+1.2597M,改进后的模型为:(?)=0.7933+1.8084M-0.1492M~2;I 值、Ca 值在整个姜瘟时间序列中始终大于0;(?)值、(?)/M 值始终大于1,这些检验结果表明姜瘟在田间呈聚集分布,但在极少数重病田当病蔸率达60%左右时,分布型呈随机分布,病蔸率达75%以上时,则趋均匀分布.抽样方法比较结果是以采用平行线、棋盘式和 Z 字型取样法为宜.文中还讨论了在不同发病率条件下的理论抽样数.     

美洲斑潜蝇空间分布型研究

通过采用聚集度指标的方法,对美洲斑潜蝇在茄子、豆角田空间分布起先了系统研究,测定结果呈聚集分布,为田间调查及防治提供了依据。     

分葱田甜菜夜蛾空间分布型及抽样技术初步研究

应用聚集指标、Iwao回归法、Taylor幂法则对分葱田甜菜夜蛾的空间分布型进行测定。结果表明甜菜夜蛾卵在分葱田呈均匀分布或聚集分布,幼虫呈聚集分布,分布的基本成分是个体群,且个体间相互吸引。并探讨了甜菜夜蛾在田间的聚集原因。通过不同抽样方式比较,Z字形抽样方式最佳。在空间分布型研究的基础上,提出了理论抽样数。     

大栗鳃金龟幼虫空间分布型研究

通过采用空间分布型统计分析方法对大栗鳃金龟3龄幼虫的分布格局进行研究,结果表明：大栗鳃金龟3龄幼虫在0～10 cm土壤中呈聚集分布,分布的基本单位是个体群,为田间调查及防治提供了科学依据。     

籼-粳杂交稻稻曲病空间分布和抽样技术研究

本文应用聚集度指标法、Iwao法和Taylor幂法则等方法,测定了籼-粳杂交稻稻曲病在田间的空间分布型和抽样技术。结果表明,稻曲病田间空间分布主要为非随机性的聚集型分布。m~*-x回归分析表明,稻曲病空间分布的基本成分是个体群,个体间相互吸引。Taylor幂法则分析显示,稻曲病病株聚集度依赖于密度。不同取样方法准确度比较结果指出,稻曲病田间抽样调查宜采用平行跳跃式取样,在此基础上提出了稻曲病田间理论抽样公式和序贯抽样表。     

稻曲病田间分布型及抽样技术初探

近年来稻曲病的危害越来越重,为探讨其田间可靠抽样方法及产量损失估算准确性,作者于1987年和1988年在晚稻黄熟期对稻曲病的田间分布型及抽样技术进行了研究。分布型采用聚块指标(M/M),群聚均数(λ)、K 值指数等聚集度指标及 Iwao 的 M—M 回归和 Tay-lor 幂的法则测定。结果表明,当稻曲病发病较轻时分布属聚集型,当发病较重时(病穗率大于45.8%),分布趋于随机型。抽样技术采用平行跳跃式、对角线式、棋盘式三种方法进行     

土壤大丽轮枝菌微菌核的快速定量检测

 微菌核是大丽轮枝菌在土壤中的主要存活结构和黄萎病的初侵染来源。对土壤中大丽轮枝菌微菌核进行定量是黄萎病监测和预警的基础。本研究以大丽轮枝菌Internal Transcribed Spacer (ITS)区特异性引物对P1/P2扩增产物的重组质粒为标准品,构建SYBR Green I实时荧光定量PCR反应的标准曲线,结合土样水筛法建立了土壤大丽轮枝菌微菌核定量检测体系。同时,建立了土壤中微菌核数量与棉花黄萎病发病率的关系模型。结果表明,实时定量PCR检测灵敏度比常规PCR高10倍,检测下限为1个微菌核/克土,在5.54×10²～5.54×10⁷copies范围内,DNA拷贝数的对数值与Ct值具有良好的线性关系。建立的土壤中微菌核个数n与Ct值之间的关系为n=e^7.3-Ct/3.905。温室人工接种微菌核数量与棉花黄萎病发病率间的线性关系为y=2.710n+0.251。     

Spatial Patterns of Microsclerotia of Verticillium dahliae in Soil and Verticillium Wilt of Cauliflower

ABSTRACT The spatial patterns of microsclerotia of Verticillium dahliae in soil and wilt symptoms on cauliflower were determined at three sites in each of two fields in 1994 and 1995. Each site was an 8 x 8 grid divided into 64 contiguous quadrats (2 by 2 m each). Soil samples were collected to a depth of 15 cm with a probe (2.5 cm in diameter), and samples from four sites in each quadrat were bulked. Plants in each quadrat were cut transversely, and the number of plants with vascular discoloration and the number without discoloration were recorded. The soil was assayed for microsclerotia by the modified Anderson sampler technique. Lloyd's index of patchiness (LIP) was used as an indicator to evaluate the aggregation of microsclerotia in the field. Spatial autocorrelation and geostatistical analyses were also used to assess the autocorrelation of microsclerotia among quadrats. The LIP for microsclerotia was greater than 1, indicating aggregation of propagules; however, the degree of aggregation at most sites was not high. Significant autocorrelation within or across rows was detected in some spatial autocorrelograms of propagules, and anisotropic patterns were also detected in some oriented semivariograms from geostatistical analyses for microsclerotia, indicating the influence of bed preparation in the fields on pathogen distribution. The parameter estimates p and theta in the beta-binomial distribution and the index of dispersion (D) associated with the distribution were used to assess the aggregation of diseased plants at each site. A random pattern of wilt incidence was detected at 7 of 12 sites, and an aggregated pattern was detected at 5 of 12 sites. The degree of aggregation was not high. A regular pattern of wilt severity was detected at all sites. The high disease incidence (77 to 98%) observed at 11 of the 12 sites could be explained by high inoculum density.     

Detection of Colletotrichum coccodes and Helminthosporium solani in soils by bioassay

The sensitivity of a bioassay in detecting soil inoculum of Colletotrichum coccodes and Helminthosporium solani was examined using potato minitubers and microplants. Tests were conducted on soils which were collected from fields in which the interval after a previous potato crop differed, and which were also artificially infested with conidia or microsclerotia. For C. coccodes , determining plant infection based on the occurrence of infected roots after 9–12 weeks was a sensitive method for detecting and quantifying the amount of inoculum in soil. Infestations of less than 0·4 microsclerotia per g soil were detected in artificially infested soils. A semiselective medium, developed for isolating C. gloeosporioides from pepper, detected soil infestations by C. coccodes as low as nine conidia or one microsclerotium per g soil in artificially infested soil. For H. solani , infection on minitubers was a sensitive measure, with soil inoculum of fewer than 10 conidia per g soil being detected. Soil infestation could be quantified by assessing the percentage surface area of minitubers covered by sporulating lesions, which was strongly related to the amount of soil infestation. The results of these bioassay tests were compared with published results for real-time quantitative PCR assays on the same soils. The two methods were in good agreement in artificially infested soils, but the bioassay appeared to be more sensitive with naturally infested soils.     

The relationship between soil inoculum density and plant infection as a basis for a quantitative bioassay of Verticillium dahliae

Using potato, eggplant and thorn apple as test plants, the relationship between soil inoculum density and plant infection was studied as a basis for the development of a quantitative bioassay of Verticillium dahliae. A linear relationship was demonstrated (P < 0.05) between soil inoculum density and population density on roots for all three test plants and for soil inoculum density and population density in sap extracted from stems for eggplant. Correlation coefficients were higher with densities on or in roots (R² varying from 0.45 to 0.99) than with densities in stems (R² varying from 0.04 to 0.26). With eggplant, population densities on/in root and in sap extracted from stems were significantly correlated at 20 and 25°C with Pearson's correlation coefficients of 0.41 and 0.53, respectively. For potato, root colonization was higher at 15 than at 20°C, whereas the reverse applied to eggplant. Stems of potato were less colonized than stems of eggplant. The pathozone sensu Gilligan (1985) was calculated to be <300 µm, indicating that infection was caused by microsclerotia which were located close to the roots. To assess the density of V. dahliae in plant tissue pipetting infested plant sap on solidified ethanol agar medium without salts yielded higher densities than using pectate medium or mixing sap with molten agar. A bioassay for determining effects of (a)biotic factors on development of V. dahliae in the plant is recommended with eggplants as a test plant, grown in soil infested with 300 single, viable microsclerotia g^-1 soil at a matric potential of –6.2 kPa, and incubated at 20°C for 8 weeks.     

Relationships Between Verticillium dahliae Inoculum Density and Wilt Incidence, Severity, and Growth of Cauliflower

ABSTRACT Microplot and field experiments were conducted to evaluate the effects of inoculum density on Verticillium wilt and cauliflower growth. Soil containing Verticillium dahliae microsclerotia was mixed with various proportions of fumigated soil to establish different inoculum densities (fumigated soil was used as the noninfested control). Seven inoculum density treatments replicated four times were established, and the treatments were arranged in a randomized complete block design. Soil was collected from each microplot immediately after soil infestation for V. dahliae assay by plating onto sodium polypectate agar (NP-10) selective medium using the Anderson sampler technique. Five-week-old cauliflower was transplanted into two beds within each 1.2- by 1.2-m microplot. At the same time, several extra plants were also transplanted at the edge of each bed for destructive sampling to examine the disease onset (vascular discoloration) after planting. Cauliflower plants were monitored for Verticillium wilt development. Stomatal resistance in two visually healthy upper and two lower, diseased leaves in each microplot was measured three times at weekly intervals after initial wilt symptoms occurred. At maturity, all plants were uprooted, washed free of soil, and wilt incidence and severity, plant height, number of leaves, and dry weights of leaves and roots were determined. The higher the inoculum density, the earlier was disease onset. A density of 4 microsclerotia per g of dry soil caused 16% wilt incidence, but about 10 microsclerotia per g of soil caused 50% wilt incidence. Both wilt incidence and severity increased with increasing inoculum density up to about 20 microsclerotia per g of soil, and additional inoculum did not result in significantly higher disease incidence and severity. A negative exponential model described the disease relationships to inoculum levels under both microplot and field conditions. Stomatal resistance of diseased leaves was significantly higher at higher inoculum densities; in healthy leaves, however, no treatment differences occurred. The height, number of leaves, and dry weights of leaves and roots of plants in the fumigated control were significantly higher than in infested treatments, but the effects of inoculum density treatments were variable between years. Timing of cauliflower infection, crop physiological processes related to hydraulic conductance, and wilt intensity (incidence and severity) were thus affected by the inoculum density. Verticillium wilt management methods used in cauliflower should reduce inoculum density to less than four micro-sclerotia per g of soil to produce crops with the fewest number of infected plants.     

Effects of crop rotation and removal of crop debris on the soil population of two isolates of Verticillium dahliae

Microsclerotia of Verticillium dahliae are produced in large numbers on senescing parts of host plants and remain viable in the soil for many years. Changes in the population density, i.e. density of microsclerotia, in the soil were measured in micro-plots using two isolates of V. dahtiae , specific to either field bean or potato, several crop sequences comprising potato, field beans and barley, and either the removal of aerial debris of the crops or incorporation into soil.
Potato was more susceptible to the potato isolate and field bean more susceptible to the field bean isolate. Removal of debris of potato and field bean reduced numbers of microsclerotia in the soil in the subsequent years, but removal of barley straw had no effect. Initially non-infested control micro-plots became infested, probably by the growth of potato roots into the naturally infested subsoil. The rate of increase of the microsclerotial population in the non-infested control micro-plots was larger than in the initially infested treatments, because more colonized debris was produced. It is concluded that removal of aerial debris of host crops is important to reduce the soil population of V. dahtiae.     

Theoretical approach to the dynamics of the inoculum density of Verticillium dahliae in the soil: first test of a simple model

A mathematical equation was developed that describes the inoculum densities of microsclerotia of Verticillium dahliae in the soil over a long time span. The equation was based on measurable parameters and ecologically meaningful principles. In the model, the number of systemic infections of plant roots during crop growth was related to soil inoculum density. In turn, formation of microsclerotia in debris and reduction of the amount of crop growth were related to the number of systemic infections. Finally, a gradual release and mortality of microsclerotia in the soil were included to calculate subsequent inoculum densities in the soil.
Fitting the function to experimental data of potato cvs Element, Ostara, Mirka and Astarte, flax, pea, barley, sugar beet, onion and faba bean gave a very high correlation between observed and predicted soil inoculum densities. The clear differences in inoculum production among potato cultivars and other crops were expressed in quantitative terms. The highest inoculum density after incorporation of the debris of a susceptible crop was estimated to occur at 2.3 thermal time units of 3600 degree days (base 0°C). Ten per cent of the initial input of inoculum was still present after 4.5 thermal time units. The model was used to predict the dynamics of soil inoculum densities for V. dahliae under various cropping frequency schemes and performed satisfactorily.     

Effect of host and inoculum patterns on take-all disease of wheat incidence,severity and disease gradient

The importance of the spatial aspect of epidemics has been recognized from the outset of plant disease epidemiology. The objective of this study was to determine if the host spatial structure influenced the spatio-temporal development of take-all disease of wheat, depending on the inoculum spatial structure. Three sowing patterns of wheat (broadcast sowing, line sowing and sowing in hills) and three patterns of inoculum (uniform, aggregated and natural infestation) were tested in a field experiment, repeated over 2 years. Disease (severity, root disease incidence, plant disease incidence and, when applicable, line and hill incidences) was assessed seven times during the course of each season and the spatial pattern was characterized with incidence-incidence relationships. In the naturally infested plots, disease levels at all measurement scales were significantly higher in plots sown in hills, compared to plots sown in line, which were in turn significantly more diseased than plots with broadcast sowing. Disease aggregation within roots and plants was stronger in line and hill sowing than in broadcast sowing. Analysis of the disease gradient in the artificially infested plots showed that the disease intensified (local increase of disease level) more than it extensified (spatial spread of the disease), the effect of the introduced inoculum was reduced by 95% at a distance of 15 cm away from the point of infestation. Yield was not significantly affected by sowing pattern or artificial infestation.     

Control of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> at a strawberry nursery by paddy-upland rotation

Effects of crop rotation between rice paddy fields and strawberry nurseries on the control of Verticillium wilt of strawberry were studied. For detecting Verticillium dahliae, the causal agent of Verticillium wilt, in soil, eggplant was used as an indicator plant. We were thus able to detect as low as 1 microsclerotium/g dry soil. In field surveys of Chiba and Hokkaido from 2000 to 2003, V. dahliae was detected in 9 of 10 upland fields but in none of 21 paddy-upland fields. In Hokkaido during 2000–2007, strawberry mother plants were planted, and plantlets were produced in upland and paddy-upland fields to assess V. dahliae infestation. Verticillium wilt of strawberry had never occurred in 72 tested paddy-upland fields, compared to 13.2–73.9% of plantlets infected with V. dahliae in upland fields. In a pot experiment in a greenhouse, two flooding treatments or two paddy rice cultivations suppressed Verticillium wilt symptoms on eggplant. In field experiments, one paddy rice cultivation in Chiba and two in Hokkaido prevented development of Verticillium wilt symptoms on eggplant. Verticillium wilt of strawberry was controlled completely with one paddy rice cultivation in infested fields in Chiba. In these field experiments, the number of microsclerotia of V. dahliae decreased under the flooding conditions for paddy rice cultivation. Based on the reduction in microsclerotia, a crop rotation system with paddy rice for 3 years (three times), green manure for 1 year, and strawberry nursery for 1 year was designed for Hokkaido.     

一种新的棉花黄萎病快速接种技术及其在病原菌致病力和寄主抗病性鉴定上的应用

 棉花品种抗黄萎病鉴定一般在田间病圃中进行,其结果受病圃中病原菌分布均匀程度、气象等因素影响极大,往往导致鉴定结果不准确。为了使鉴定方法简单、科学、可靠,我们在温室条件下比较了3种苗期接种棉花黄萎病的方法,即切根蘸孢子法(接种浓度为10⁶分生孢子/mL);菌培养物土壤接种法(0.5%、1%、2%,w/w);微菌核土壤接种法(10³个微菌核/g土)。结果表明,切根蘸孢子法导致棉苗发病均匀、严重、迅速,播种35~45 d后即可得到均匀一致的发病结果。而其它2种接种方法在播种75 d后才得到相对稳定的发病结果。同时,研究还表明接种浓度为10⁴分生孢子/mL所导致的黄萎病显著比10⁵或10⁶的轻。利用切根蘸孢子法在室内鉴定12个棉花品种或品系的抗黄萎病能力,证明该方法是抗黄萎病快速鉴定的有效方法。此外,该鉴定方法还可快速鉴定黄萎病菌不同菌株的致病性,并可应用于作物对其它土传病害的抗病性鉴定上。