Studies on the biology and control of cavity spot of carrots

Using a selective medium of corn meal agar with pimaricin and rifamycin, isolations from asymptomatic periderm of carrots grown on experimental plots or in commercial crops most frequently yielded the fast-growing species Pythium intermedium, P. sylvaticum or P. ultimum. In contrast isolations from cavity spot lesions mostly produced the slow-growing species P. violae and P. sulcatum. Following treatment of crops with metalaxyl + mancozeb, few isolations from asymptomatic periderm produced Pythium spp. and generally there was a reduction in the number of cavities. Treatment had little or no effect on size distribution of cavities. Cavity spot incidence was significantly less at higher pH values, fields of pH 8·0 and above producing carrots with little or no disease.     

Detection of Pythium violae and Pythium sulcatum in carrots with cavity spot using competition ELISA

Conventional methods indicated that Pythium violae was most commonly isolated from carrot cavity spot samples from 14 UK sites. For one site the most frequently isolated species was Pythium sulcatum. Results of similar isolation work were compared with the assay of cavity spot lesions using polyclonal antibodies, raised to P. violae or P. sulcatum, in competition ELISA. Where lesions were artificially induced the test confirmed which pathogen was causal. With cavities developed on the field-grown carrots P. violae again predominated and the ELISA confirmed this. In one sample P. sulcatum was also isolated from a small number of lesions and was not detected in ELISA. The competition ELISA did not indicate presence of either Pythium in a range of non-cavity spot lesions from which attempts at isolation were negative.     

Biochemical characterisation of Pythium spp. involved in cavity spot of carrots in France

The pathogenicity and growth rate in vivo were assessed on 27 isolates of Pythium spp. recovered from cavity spot lesions on carrots grown in various parts of northwest France. Polyacrylamide gel electrophoresis of isoesterases was used to identify the Pythium spp. involved. Slow-growing isolates were more aggressive than fast-growing ones when inoculated on carrot tap roots. Isoesterase patterns identified the slow-growing isolates as P. violae and P. sulcatum; P. ultimum and P. intermedium were identified among the less aggressive fast-growing isolate group, in which some isolates were also classed as P. sylvaticum or P. irregulare, which have similar electrophoretic profiles. The incidence of Pythium spp. associated with the disease in France is discussed in regard to cavity spot in other countries.     

Influence of parental density on the distribution pattern of eggs in the common cabbage butterfly,Pieris rapae crucivora

To investigate the relation between the distribution pattern of eggs and the parental density in the common cabbage butterfly, Pieris rapae crucivora, the countings of egg number per plant were made on both cabbage plants cultivated in the farm and planted in the net house in which the female butterflies were released at various densities. The frequency distribution of eggs fits well to the negative binomial excepting the cases where they agree withPoisson series, and the degree of aggregation expressed as the reciprocal of the parameter, 1/k, tends to decrease as the egg or parental density increases. At the same parental density, however, the distribution of eggs can be described by the negative binomial with a common parameter, k_c, regardless of the difference in the density of laid eggs. In the case where a single female butterfly lays eggs, the spatial pattern of egg distribution is always lean, while its frequencies conform toPoisson or the negative binomial series. This lean changes toward patchy with increasing the parental density. From these results, it is concluded that the degree of aggregation in the distribution of eggs decreases with the increase of the parental density.     

The spatial distribution of the wheat-bulb fly,Leptohylemyia coarctata (fall.) (Diptera: Anthomyiidae)

The spatial distribution of the eggs, larvae, pupae and adults of the wheat-bulb fly was investigated by fitting 42 sets of data comprising 1334 samples to the Poisson and negative binomial distributions, and by using the power law (S²=am^b). In general, the tests indicated that all stages were aggregated and fitted the negative binomial model.     

Effect of soil cultivation and anaerobiosis on cavity spot of carrots

Cultivating between rows and narrow beds of carrots reduced the severity of cavity spot in one year and reduced the incidence and severity in single rows in a second year on land where the disorder was endemic. Growing carrots on ridges initially reduced symptoms more than inter-row cultivation. Soluble carbohydrates leached from carrot roots in vitro and the quantities increased when anaerobic conditions were imposed. Untreated and cell-free extracts of soil suspensions amended with 1% glucose and incubated anaerobically applied to carrots in the laboratory caused the outer layers of the secondary phloem to collapse resembling a cavity spot lesion. Unamended and fresh soil solutions had no effect.     

Experimental studies on the distribution of the aphid counts

The present paper dealt with the sequential changes of the distribution pattern of apterous females aphid populations, that were artificially settled at the beginning on the experimental barley ‘field'. The aphids were settled at random or even with a fixed denisty per plant. For five or six days after the settling, the number of individuals followed the negative binomial distributions in all cases while the parameters k and p were varying. The estimated values of k were rather small for the first one week after the settling, which may suggest that the number of moving aphids between plants was relatively small and the degree of concentration expressing the intrinsic increase was high. After that, as the number of individuals increased, the number of moving aphids between plants would be considered to be increased. It was found that with the lapse of time the degree of concentration decreased or k became larger. The distribution of aphids per blade in a plant was also described briefly.     

Cavity spot of carrots: Interactions between the host and pathogen, related to the cell wall

This study investigates the structural aspects of cavity spot pathogenesis. Different Pythium spp. isolated from infected carrots, apples and melons were cultured on agar in Petri dishes and used for inoculation of uninfected carrots. Only slow-growing Pythium spp. (< 15 mm day^-1), such as P. violae and P. sulcatum caused cavity spot lesions. It is suggested that slow-growing species are able to penetrate, albeit slowly, into the plant tissue for 3 to 4 days before a hypersensitive reaction develops. Fast-growing species, however, did not cause lesions. Based on ultrastructural observations, we suggest that the following sequence of events occurs between the plant and the pathogen: The fungus infects the walls and grows for several days, during which time small amounts of wall-degrading enzymes are secreted. Phenylalanine ammonia lyase (PAL) activity and phenols increase linearly immediately upon inoculation. There was a lag phase of about 5 days before lignin began to increase linearly for about a month. Dissolution of wall components decreases the solute potential and water potential in the apoplast. Thus, water moves from the symplast into the apoplast, the turgor pressure gradually dissipates, and the cells shrink and eventually die.     

Process generating the ditribution pattern of eggs of the common cabbage butterfly,Pieris rapae crucivora

The process generating the negative binomial in the distribution pattern of eggs of the common cabbage butterfly, Pieris rapae crucivora, was investigated by releasing the female adults in a net house where cabbages were planted. The distribution of butterflies visited and laid an egg or more per plant followed thePoisson series under the uniform light condition, while that of eggs laid per visit conformed to the logarithmic distribution. From these results, it may be concluded that the negative binomial arises from compounding of thePoisson and the logarithmic distribution. The observed frequency of eggs found per plant fitted to the negative binomial with parameter thus computed theoretically. The change in the degree of aggregation with the increase of the parental density was considered in connection with the above results.     

Evaluation of statistical precision and design of efficient sampling for the population estimation based on frequency of occurrence

The binomial sampling to estimate population density of an organism based simply upon the frequency of its occurrence among sampled quadrats is a labour-saving technique which is potentially useful for small animals like insects and has actually been applied occasionally to studies of their populations. The present study provides a theoretical basis for this convenient technique, which makes it statistically reliable and tolerable for consistent use in intensive as well as preliminary population censuses. Firs, the magnitude of sampling error in relation to sample size is formulated mathematically for the estimate to be obtained by this indirect method of census, using either of the two popular models relating frequency of occurrence (p) to mean density (m), i.e. the negative binomial model, p=1−(1+m/k)^−k, and the empirical model, p=1−exp(−am^b). Then, the equations to calculate sample size and census cost that are necessary to attain a given desired level of precision in the estimation are derived for both models. A notable feature of the relationship of necessary sample size (or census cost) to mean density in the frequency method, in constrast to that in the ordinary census, is that it shows a concave curve which tends to rise sharply not only towards lower but also towards higher levels of density. These theoretical results make it also possible to design sequential estimation procedures based on this convenient census technique, which may enable us with the least necessary cost to get a series of population estimates with the desired precision level. Examples are presented to explain how to apply these programs to acutal censuses in the field.     

植物与病原菌互作的蛋白质组学研究进展

深入认识植物与病原菌的识别方式、亲和性或非亲和性的互作模式,对于揭示植物-病原菌互作机制研究具有重要意义.利用蛋白质组学方法研究病原菌侵染植物过程,分析相关的基因和蛋白,有助于从分子水平上探究植物-病原菌相互作用机制.本文概述了植物-病原菌的互作机制,系统介绍了差异蛋白质组学分析方法在植物-病原真菌、植物-病原细菌两类互作系统中的应用,分析了植物与病原菌互作过程中可能涉及的差异表达功能蛋白,并对当前蛋白质组学技术在植物与病原菌互作研究中存在的诸多问题进行了探讨.     

Spatial distribution of the green peach aphid used in estimating the populations of gynoparae

Summary In 2 years, during the initial invasion of peach leaves by the green peach aphid,Myzus persicae (Sulzer), the number of gynoparae was low, and the distribution on leaves was random. Then as the mean number increased, the distribution became intermediate and could not be distinguished from either a Poisson or a negative binomial. Finally, as the mean continued to increase, the variance increased rapidly, and the population was found to fit a negative binomial distribution. Thus the aggregation response was verified because the dispersion pattern fitted a contagious distribution. A sampling plan was devised by which the dispersion parameterk was used to estimate the density of aphids per leaf based on the percentage of leaves infested. Sampling the third year of the study confirmed the validity of the sampling parameter that had been calculated from data for the 2 previous years.     

Microbiological differences between limed and unlimed soils and their relationship with cavity spot disease of carrots (Daucus carota L.) caused by Pythium coloratum in Western Australia

Application of lime (4000 kg ha^-1) to a soil used for commercial carrot production (pH 6.9) significantly (p<0.05) reduced the incidence of cavity spot disease of carrots compared to unlimed soil (pH 5.1). It significantly (p<0.01) increased soil microbial activity as measured by the hydrolysis of fluorescein diacetate and arginine ammonification. The application of lime resulted in a significant (p<0.01) increase in the total numbers of colony forming units (efu) of aerobic bacteria, fluorescent pseudomonads, Gram negative bacteria, actinomycetes and a significant (p<0.01) decrease in the cfu of filamentous fungi and yeasts compared to unlimed soil. Liming also increased the cfu of non-streptomycete actinomycetes rarely reported in similar studies. These non-streptomycete actinomycetes were estimated and isolated using polyvalent Streptomyces phages and the dry heat technique to reduce the dominance of streptomycetes on isolation plates. The non-streptomycete actinomycetes isolated included species of Actinoplanes, Micromonospora, Streptoverticillium, Nocardia, Rhodococcus, Microbispora, Actinomadura, Dactylosporangium and Streptosporangium. The numbers of actinomycetes antagonistic to Pythium coloratum, a causal agent of cavity spot disease of carrots increased in soil amended with lime. Application of lime also reduced the isolation frequency of P. coloratum from asymptomatic carrot roots grown in soil artificially infested with the pathogen, 3, 4 and 5 weeks after sowing.     

Cavity spot of carrots: II. Cell-wall-degrading enzymes secreted by Pythium and pathogen-related proteins produced by the root cells

This study investigates the biochemical relationships between carrot roots and Pythium violae, the pathogen responsible for cavity spot (CS) disease. P. violae isolates obtained from CS lesions, cultured in Petri dishes on agar were used for inoculation of uninfected mature carrots. The fungus secreted a wide spectrum of enzymes that degraded the cellulose and pectic substances of the carrot cell walls. Cellulase and polygalacuronase (pg) showed the highest activity during the first day post-inoculation, subsequently declining. Pectin lyase (PnL), pectate lyase (PeL) and pectin methylesterase (PME) gradually increased to their highest levels of activity 14 to 30 days post-inoculation. This pattern of activity enables the penetration of the fungus through the walls of the host cells and the establishment of the hyphae. Several plant pathogen-related substances such as peroxidase, chitinase, glucanase and polyphenol oxidase were produced in the infected tissue. Peroxidase activity rose in the inoculated roots from day 1 post-inoculation. Chitinase, glucanase and polyphenol oxidase activities first appeared 3–4 days post-inoculation. At this time, two bands corresponding to chitinase at about 26 and 33 KDa and one band corresponding to glucanase at about 24 KDa could be resolved by SDS-PAGE.