Effect of root-knot nematodes on cajanol accumulation in the vascular tissues of pigeonpea after stem inoculation with Fusarium udum

The effect of root-knot nematodes ( Meloidogyne javanica and M. incognita ) on production of the isoflavonoid phytolexin, cajanol, was investigated in pigeonpea ( Cajanus cajan ) plants infected with Fusarium udum , the causal organism of Fusarium wilt. Seven-day-old seedlings of a wilt-resistant pigeonpea cultivar, ICP 9145 and a wilt-susceptible cultivar, Malawi Local, both of which were moderately susceptible to the nematode, were grown in soil infested with 2000 Meloidogyne juyeniles per plant. A duplicate set of plants remained free from nematodes. Twenty-one days later, all the plants were inoculated with F. udum by stem puncture. Quantitative estimates of cajanol in the vascular tissues were made at intervals up to 15 days after inoculation with the fungus. No external symptoms of wilt appeared in any plants of the wilt-resistant cultivar in the absence of the nematode. However, when inoculated with the nematode, two thirds of the plants developed wilt symptoms. Cajanol levels were lower in both the wilt-resistant and wilt-susceptible plants in the presence of the nematode than in its absence, although this effect was considerably more marked in the wilt-resistant cultivar. These results indicate that the root-knot nematode is capable of breaking resistance of ICP 9145 to Fusarium wilt and that at least part of the mechanism of this effect is retarded cajanol accumulation.     

土壤中烟草根黑腐病菌的实时定量PCR检测技术研究

 Thielaviopsis basicola is a soil-borne plant pathogen which causes root rot disease in tobacco plants. Detection and monitoring of T. basicolain soil is of great significance to control this disease. Based on the differences in internal transcribed spacer (ITS) sequences of T. basicola and other fungal pathogens, a specific primer pair Tb1/Tb2 for T. basicolawas developed. The results showed that the primer pair gave a single amplicon of 330 bp from T. basicola and revealed no undesirable cross-reaction with other seven soil-borne pathogen isolates and three tobacco rhizosphere dominant fungi isolates. With a series of 10-fold genomic DNA dilutions of T. basicola, the detection limit of 1 pg/μL in conventional PCRand100 fg/μL in real-time quantitative PCR was achieved. With DNA from the soil inoculated with different numbers of T. basicola conidia, the detection limit was 10 conidia per reaction in conventional PCR and 0.4 conidia per reaction in real-time quantitative PCR.     

The occurrence in Denmark of black root rot of pea caused by Thielaviopsis basicola

Thielaviopsis basicola has been shown to be a root pathogen of pea of considerable importance in Denmark. The fungus is only found in fields with one or more previous pea crops in the field history. In the dry and warm growing season of 1989 the fungus was found in 0·6% and 3·2% of the fields in two separate areas in Denmark. In the fields where T. basicola was detected the average disease severity index in plant samples was 51·8, whereas the average disease severity index in plant samples without the fungus was 27·0. The average yield of green peas was reduced from 5167 to 4171 kg/ha when T. basicola was present. For detection and isolation of T. basicola it is important to use a technique combining microscopic examination, a semi-selective medium and a dilution plate method.     

Reassessment of the Role of Saprophytic Activity in the Ecology of Thielaviopsis basicola

ABSTRACT The ability of Thielaviopsis basicola to survive saprophytically in soil was investigated using root tissue from susceptible hosts as organic substrates. Inoculum densities were lower in soils amended with root tissue than in nonamended controls after 2 and 4 weeks of incubation. The greatest decrease occurred in soils containing the highest concentration of root tissue or in soils in which root tissue included the soluble components of the living root. Reproduction by T. basicola also was examined in axenic media containing either killed root pieces or various carbohydrates as the sole carbohydrate source. T. basicola utilized killed root tissue as a carbohydrate source in axenic media, particularly in cultures in which root tissue included the soluble components. Enzymatic activities of T. basicola, however, did not result in maceration of the root tissue. T. basicola utilized sucrose and cellobiose, but did not utilize structural carbohydrates such as cellulose, hemicellulose, or pectin. Based on the absence of significant saprophytic ability, T. basicola should be classified ecologically as an obligate parasite.     

Interactions between the biocontrol agent Pseudomonas fluorescens CHA0 and Thielaviopsis basicola in tobacco roots observed by immunofluorescence microscopy

Pseudomonas fluorescens CHA0 protects plants from damage caused by several soilborne fungi. In this work, immunofluorescence microscopy was used to investigate the colonization of tobacco roots by CHA0 and its physical relationship with the black root rot fungus Thielaviopsis basicola . The pseudomonad colonized the rhizoplane shortly after planting of tobacco seedlings in sterile soil microcosms, in which it had been introduced as soil inoculant. CHA0 was found between and inside cells in the epidermis and the cortex, as well as in the xylem vessels, within 4–7 days after planting of seedlings. The presence of CHA0 delayed the colonization of the interior of tobacco roots by T. basicola compared with the treatment in which only the fungus had been inoculated. Likewise, the pseudomonad reduced the extent of black root rot from 82% to 28%. However, CHA0 was seldom found in contact with the mycelium of T. basicola or in its vicinity, indicating that direct colonization of the mycelium of T. basicola by CHA0 was not required for protection of tobacco against black root rot. Overall, the results suggest that the interior of the root is a key site for implementation of the strain's biocontrol activity against soilborne plant-pathogenic fungi.     

Differential Sensitivity of Phytophthora parasitica var. nicotianae and Thielaviopsis basicola to Monomeric Aluminum Species

ABSTRACT Aluminum (Al) is toxic to many plant pathogens, including Thielaviopsis basicola and Phytophthora parasitica var. nicotianae. Because fungi-toxicity of Al has been described in soils over a wide pH range, multiple species of Al may be responsible for pathogen suppression. The goals of this work were to determine the sensitivity of T. basicola and P. para-sitica var. nicotianae to Al over a range of pH values, quantify the toxicity of monomeric Al species to production of sporangia of P. parasitica var. nicotianae and chlamydospores of T. basicola, and detect the accumulation of Al in pathogen structures. A complete factorial treatment design was used with Al levels ranging from 0 to 100 muM and pH levels ranging from 4 to 6 in a minimal salts medium. The chemistry of test solutions was modeled using GEOCHEM-PC. Colonies were grown in 5% carrot broth, and after 1 or 2 days, the nutrient solution was removed, colonies were rinsed with water, and Al test solutions were added to each of four replicate plates. After 2 days, propagules were counted and colonies were stained with the Al-specific, fluorescent stain lumogallion. The oomycete P. parasitica var. nicotianae was sensitive to multiple monomeric Al species, whereas sensitivity of T. basicola to Al was pH-dependent, suggesting that only Al(3+) is responsible for suppression of this fungal pathogen. Chlamydospore production by T. basicola was inhibited at pH values <5.0 and Al levels >20 muM, whereas sporangia production by P. parasitica was inhibited at Al levels as low as 2 muM across all pH values tested. The lumogallion stain was an effective technique for detection of Al in fungal tissues. Aluminum accumulated in sporangia and zoospores of P. parasitica var. nicotianae and in nonmelanized chlamy-dospores of T. basicola, but not in cell walls of either organism. The differential sensitivity of the two organisms may indicate that true fungi respond differently to Al than members of the oomycota, which are more closely related to plants.     

Effects of soil moisture and sowing depth on the development of bean plants grown in sterile soil infested by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> and <Emphasis Type="Italic">Trichoderma harzianum</Emphasis>

The effects of soil moisture (varying from 15% to 42% (v/v)) and sowing depth (1.5–6.0 cm) on the development of bean plants grown in sterile soil infested by the pathogen Rhizoctonia solani and its antagonist Trichoderma harzianum were studied under greenhouse conditions. The four possible combinations of soil infestation with both fungi were tested. Disease severity, percentage of plants emerged, plant height and dry weight were evaluated 3 weeks after sowing. Emergence rate and growth of plants inoculated only with R. solani were not affected by soil moisture, but in the presence of both fungi, plant emergence, plant height and dry weight significantly decreased when soil moisture diminished. Deep sowing significantly reduced the emergence rate and growth of those plants that were inoculated with R. solani only. However, when the soil was infested with both fungi, the effect of sowing depth was not significant. At a sowing depth of 6.0 cm, the percentage of plants emerged was 50% in the presence of T. harzianum, but only 6.7% when the pathogen was inoculated alone. The antagonist protected bean seedlings from pre-emergence damping-off, reduced disease severity and increased plant growth in the presence of R. solani, especially in moist soil.     

Effects of DL-methionine on hatching and activity of Meloidogyne incognita eggs and juveniles

Several concentrations of DL-methionine were tested in the laboratory for their effects on Meloidogyne incognita (Kofoid & White) Chitwood egg hatching and juvenile activity in aqueous suspensions and infested soil. After 7 days in methionine solutions, the proportions of hatched eggs were reduced by 23.3% at 0.25 mg litre(-1) methionine and by 76.4% at 25 g litre(-1) when compared with controls in tap water. An effect of methionine solutions on juvenile activity was also apparent after 24 h in the 25 g litre(-1) treatment, where the percentage of active M incognita juveniles was reduced by 16.3%. Further reductions in nematode mobility were observed as the time of exposure increased, and at lower methionine concentrations over longer exposure times. When methionine solutions were applied to soil infested with M incognita, reductions in egg hatching and juvenile activity were observed at 0.1 and 1 mg methionine g(-1) soil in both sand and clay-loam soils. The percentage of hatched eggs one week after methionine application at rates of 1 mg g(-1) was lower in sand (5.6%) and clay-loam soil (20.8%) than in controls where egg hatch was 52.0% and 48.0% in sand and soil, respectively. Non-active juveniles were found at 1 mg methionine g(-1) soil one week after its application to soil.     

生姜根结线虫病原鉴定及发生规律

采用田间调查、接种试验、电镜与显微镜观察以及酯酶同工酶电泳等方法,对引发生姜癞皮病的病原及发生规律进行了研究.结果表明,引起生姜癞皮病的病原为南方根结线虫Meloidogyne incognita.该病在每年6月中旬开始发生,8、9月份危害严重.病原在生姜上一年可发生完整的4代,完成1代平均约需35天.病原主要在0～40cm的土层内分布和危害,但具体分布情况依寄主生长状况而稍有差异.南方根结线虫繁殖速率受初始接种密度的影响也很大,当初始接种密度较低时,线虫繁殖速率较高,初始接种密度增大,繁殖速率降低,其平衡密度为每100g干土746.20个卵.     

Inoculum Densities of Fusarium oxysporum f. sp. vasinfectum and Meloidogyne incognita in Relation to the Development of Fusarium Wilt and the Phenology of Cotton Plants (Gossypium hirsutum)

ABSTRACT Development of Fusarium wilt in upland cotton (Gossypium hirsutum) usually requires infections of plants by both Meloidogyne incognita and Fusarium oxysporum f. sp. vasinfectum. In this study, the soil densities of M. incognita and F. oxysporum f. sp. vasinfectum and the incidence of Fusarium wilt in three field sites were determined in 1982-1984. Multiple regression analysis of percent incidence of Fusarium wilt symptoms on population densities of M. incognita and F. oxysporum f. sp. vasinfectum yielded a significant fit (R (2) = 0.64) only on F. oxysporum f. sp. vasinfectum. Significant t-values for slope were also obtained for the interaction of M. incognita and F. oxysporum f. sp. vasinfectum, but densities of M. incognita and F. oxysporum f. sp. vasinfectum were also related on a log(10) scale. The physiological time of appearance of first foliar symptoms of Fusarium wilt, based on a degree-days threshold of 11.9 degrees C (53.5 degrees F), was used as a basis for determining disease progress curves and the phenology of cotton plant growth and development. Effects of Fusarium wilt on plant height and boll set were determined in three successive years. Increases in both of these plant characteristics decreased or stopped before foliar symptoms were apparent. Seed cotton yields of plant cohorts that developed foliar wilt symptoms early in the season (before 2,000 F degree-days) were variable but not much different in these years. This contrasted with cohorts of plants that first showed foliar symptoms late in the season (after 2,400 F degree-days) and cohorts of plants that showed no foliar symptoms of wilt. Regression analyses for 1982-1984 indicated moderate to weak correlations (r = 0.16-0.74) of the time of appearance of the first foliar symptoms and seed cotton yields.     

Occurrence of Polyscytalum pustulans, Phoma foveata and Fusarium solani var. coemleum in field soils in Scotland

The effectiveness of various methods for detecting three fungal potato pathogens was compared with artificially infested soil, naturally infested tuber-borne soil and field soil. In the spring of 1985 and 1986 field soils from 30 farms in north-east Scotland were sampled just before planting a seed potato crop and 6 months after harvesting such a crop. The minimum statutory gap between crops is 5 years. Polyscytalum pustulans was recovered from 32 out of 60 field soil samples taken 6 months after harvest while from fields sampled in the spring before a potato crop was planted the fungus was isolated from 10 out of 30 soils in 1985 and five out of 30 in 1986. Phoma foveata was isolated from only one out of 60 pre-planting soil samples but Fusarium solani var. coeruleum was recovered from eight of these soils.
Microplant bait plants were grown over 3 years at an experimental farm near Edinburgh in various fields at different intervals after a previous potato crop. Contamination by P . pustulans was not related to interval after potatoes between 1 - 7 years. No contamination was recorded in fields where potatoes had not been grown for more than 30 years.     

Comparison of Soil Receptivity to Thielaviopsis basicola, Aphanomyces euteiches, and Fusarium solani f. sp. pisi Causing Root Rot in Pea

ABSTRACT Soil receptivity as a quantifiable characteristic ranging from conduciveness to suppressiveness to soilborne pea pathogens Thielaviopsis basicola and Aphanomyces euteiches was determined by analysis of differences in disease response curves obtained by artificial introduction of inoculum into natural field soil samples. Several parameters, including maximum root rot severity, the area under the health index curve, scores on the first axis of a principal component analysis (PCA) on dose responses, and Weibull model fitting were used to describe the disease responses. In all cases, the Weibull model gave satisfactory fits. PCA yielded a first axis that comprised 86% of the variance found when using Weibull predicted responses for T. basicola and 74% of the variance found for A. euteiches. This PCA axis essentially represented the average increase in disease severity due to the addition of increasing doses of inoculum to the soil. The Weibull scale parameter B, which represents the amount of inoculum necessary to increase root rot severity by 63% with respect to the level caused by pathogens naturally present in the soil, is another means of quantifying the receptivity of soils to these plant pathogens. Weibull parameter B, maximum root rot severity, the areaunder the health index curve, and the scores on the first PCA axis were strongly correlated for each of the pathogens tested individually. To compare the extent and behavior of soil receptivity responses to different pathogens, Weibull parameters B and C (slope at dose B) were chosen because of their universal definition, in contrast to PCA scores. Comparison of the average levels of Weibull parameters B and C indicated significant differences between the pathogens. Yet, no significant similarity in the ranking of the soils was found for the three pathogens, demonstrating that individual soils may interact with different pathogens in totally different ways. In general, soils were suppressive to T. basicola but conducive to A. euteiches, whereas their response to Fusarium solani f. sp. pisi ranged from conducive to suppressive. Therefore, risk assessment of soils prior to planting may require different strategies for each pathogen. Bioassays with soil samples taken before the last pea crop in 1987 and 1991 revealed a significant increase in the natural inoculum potential of soils that mainly was accounted for by A. euteiches and Pythium spp. These results strongly indicate that A. euteiches must be considered one of the most threatening pathogens to pea crops in the Netherlands.     

Aerial Transmission of Thielaviopsis basicola, a Pathogen of Corn-Salad, by Adult Shore Flies

ABSTRACT Chlamydospores of Thielaviopsis basicola were consistently observed in frass excreted by adults and larvae of shore flies that were collected in the immediate vicinity of naturally infected corn-salad plants obtained from a commercial greenhouse production facility. Approximately 95% of the adult flies and 85% of the larvae were internally infested with the pathogen. Pathogen-free adult shore flies were subsequently shown to acquire the pathogen by ingestion after feeding on naturally infected plants. Viable propagules of the pathogen were excreted by these internally infested adults and were capable of transmitting the pathogen to healthy seedlings, which subsequently became infected.     

Rhizosphere Competence of Pythium oligandrum

ABSTRACT The associations of Pythium oligandrum with the root cortex, rhizoplane, and rhizosphere were measured with 11 crop species. This work was expedited by the use of a semiselective technique for isolation of P. oligandrum from soil and plant material. Cortical colonization of roots by P. oligandrum was not detected, and the rhizoplanes of the roots of most crops were free of the fungus. However, P. oligandrum was detected in large quantities with every crop tested when roots with adhering soil (rhizosphere soil) were assayed. Different crop species and cultivars of cantaloupe, cauliflower, and tomato varied in rhizosphere densities of P. oligandrum, but rhizosphere population densities of the fungus were consistently higher than in nonrhizosphere soils with plants grown in P. oligandrum-infested sterilized potting mix or an unsterilized mineral soil. After transplanting tomatoes into potting mix infested with P. oligandrum, increases in CFU occurred over time in the rhizosphere but not in the nonrhizosphere soil. In trials on delivery methods of inoculum of P. oligandrum, the rhizosphere populations of tomato plants grown in potting mix were about sixfold higher compared to seed-coat treatments when ground, alginate pelleted oospores were applied to seedlings growing in plug containers prior to transplanting or to pots containing potting mix before direct seeding.     

Use of crop residues for the control of Meloidogyne incognita under laboratory conditions

This laboratory study evaluates the biofumigant effect of different organic materials with the aim of developing non-chemical alternatives for the management of Meloidogyne incognita (Kofoid & White) Chitwood populations. Sources of organic material from the production system were selected with the aim of reducing agricultural residue accumulation problems as well as decreasing the costs due to the use of chemical fertilizers and pesticides. The selected materials were residues from pepper, strawberry, tomato and cucumber crops, orange juice industry residues, commercial manure and sheep manure, applied at different dosages. Two biofumigation assays were performed under laboratory conditions, using alkaline soils from the Torreblanca area (Murcia, Spain) and acidic soils from the Villa del Prado area (Madrid, Spain). The assays evaluated the effect of the treatments on M. incognita juveniles and other soil organisms, the nematode galling index on tomato roots (susceptible cv. Marmande) grown in the biofumigated soil and soil fertility parameters. The results showed that all biofumigant materials significantly decreased M. incognita populations and galling indices in tomato cv. Marmande. A greater effect was observed on galling indices when applying crop residues together with manure than with the residues alone. Biofumigation had a general beneficial effect on soil fertility, generally increasing nitrogen, organic carbon, pH and potassium levels, and also calcium levels when crop residues of pepper and strawberry were applied. There were no important variations in the number of saprophagous nematodes, dorylaimids and enchytraeids.     

象耳豆根结线虫的PCR鉴定和检测方法

 象耳豆根结线虫是一种在中国具有潜在经济重要性的农作物病原物。为提供有助于控制象耳豆根结线虫传播扩散的方法,研制了该线虫的快速PCR鉴定和检测法。该方法PCR引物的扩增目标为rDNA-IGS2区域,其设计依据象耳豆根结线虫与南方、爪哇、花生和北方根结线虫在该区域核酸序列的差异。通过对6种近似根结线虫的不同地理群体及自然土壤线虫群体的测试,验证了设计的PCR引物针对象耳豆根结线虫的特异性和可靠性。本方法具有快速灵敏的特点,可用于象耳豆根结线虫单条线虫的直接鉴定以及混合土壤线虫群体中象耳豆根结线虫的检测。     

Host–parasite relationships of Meloidogyne incognita on spinach

Host–parasite relationships in root-knot disease of spinach caused by Meloidogyne incognita race 1 were studied under glasshouse conditions. Nematode-induced mature galls were large and usually contained one or more females and egg masses with eggs. Feeding sites were characterized by the development of giant cells containing granular cytoplasm and many hypertrophied nuclei. The cytoplasm in these giant cells was aggregated alongside the thickened cell walls. Stelar tissues within galls appeared disorganized. The relationship between initial nematode population density ( P _i) in a series from 0–128 eggs and second-stage juveniles per cm³ soil and growth of spinach cv. Symphony F₁ seedlings was tested under glasshouse conditions. A Seinhorst model [ y = m  + (1 −  m ) z ^P–T ] was fitted to fresh top- and total plant-weight data for inoculated and control plants. Tolerance limits ( T ) of spinach cv. Symphony F₁ to M. incognita race 1 for fresh top and total plant weights were 0·25 and 0·5 eggs and second-stage juveniles per cm³ soil, respectively. The minimum relative values for fresh top and total plant weights were zero in both cases at P _i ≥ 32 eggs and second-stage juveniles per cm³ soil. Root galling was least at low initial population densities and greatest at 16 eggs and second-stage juveniles per cm³ soil. Maximum nematode reproduction rate was 33·1-fold at the lowest P _i.     

AM真菌诱导的番茄信号物质及其对根结线虫的 抑制 效应

丛枝菌根(AM)真菌能诱导植物合成一些信号物质,如茉莉酸(JA)、水杨酸(SA)、一氧化氮(NO)、H2O2等,这些信号在AM真菌与植物识别、共生体建立和激活植物防御系统过程中发挥着重要作用[1].     

Development of a new management strategy for the control of root-knot nematodes (Meloidogyne spp) in organic vegetable production

The nematophagous fungus, Pochonia chlamydosporia (Goddard) Zare & Gams, has been investigated as a potential biological control agent for use in integrated pest management strategies for Meloidogyne incognita (Kof & White) Chitwood in vegetable crops. The release of the fungus as a biological control agent requires a diagnostic method to monitor its spread in the environment and to gain knowledge of its ecology. Only molecular methods are sufficiently discriminating to enable the detection of specific isolates of fungi in soil. A method to extract DNA from soil was developed to increase the efficacy of PCR-based diagnostic tests that use specific primers. A selected isolate of P chlamydosporia var catenulata was applied at densities similar to those that occur naturally in nematode-suppressive soils. The fungus significantly reduced nematode infestations in soil following a tomato crop, in a strategy that combined the use of the fungus with crop rotation. The survival of the fungus in soil was also examined in controlled conditions in which it remained in soil in densities significantly greater than its original application rate for at least 5 months. Hence, it seems that populations of this fungus may be built up in soil and have significant effects on the regulation of root-knot nematode populations.     

Rice nematode problems in Nigeria: Their occurrence,distribution and pathogenesis

Abstract

In a survey of plant parasitic nematodes associated with or affecting rice throughout Nigeria, some important nematode pests, especially the white tip disease nematode, Aphelenchoides besseyi and the rice root nematodes Hirschmanniella spinicaudata and H. oryzae were identified from seed, soil and root samples from swamp rice fields respectively. The sugarcane cyst nematode, Heterodera sacchari occurred in swamp rice fields only around the major sugarcane estates in Nigeria. The root‐knot Meloidogyne incognita and the root lesion nematode, Pratylenchus brachyurus were also encountered in upland (rainfed) rice fields. The white tip disease nematode, A. besseyi occurred at low levels in soils and rice seeds throughout the country. High population levels of H. spinicaudata and H. oryzae were encountered especially in areas where monoculture of rice is practised. General chlorosis, poor tillering and significantly reduced yield have been observed due to H. spinicaudata. Rice plants attacked by H. sacchari also showed intense chlorosis, delayed and reduced tillering and reduced grain yield. The roots of attacked plants were twiggy, very necrotic and blackened. The root‐knot M. incognita and the root lesion nematodes P. brachyurus have both been observed to reduce rice yields. Rice cultivars screened for reactions to the nematodes showed varying degrees of susceptibilities. Some varieties were however resistant to the root‐knot nematode, M. incognita.