Pathogenicity and host‐parasite relationships of the root‐knot nematode Meloidogyne incognita on celery

Pathogenicity and host‐parasite relationships in root‐knot disease of celery (Apium graveolens ) caused by Meloidogyne incognita race 1 were studied under glasshouse conditions. Naturally and artificially infected celery cv. D’elne plants showed severe yellowing and stunting, with heavily deformed and damaged root systems. Nematode‐induced mature galls were spherical and/or ellipsoidal and commonly contained more than one female, males and egg masses with eggs. Feeding sites were characterized by the development of giant cells that contained granular cytoplasm and many hypertrophied nuclei. The cytoplasm of giant cells was aggregated along their thickened cell walls and consequently the vascular tissues within galls appeared disrupted and disorganized. The relationship between initial nematode population density (Pi) and growth of celery plants was tested in glasshouse experiments with inoculum levels that varied from 0 to 512 eggs and second‐stage juveniles (J₂) mL^?1 soil. Seinhorst's model y = m + (1 – m)z^P–T was fitted to height and top fresh weight data of the inoculated and control plants. The tolerance limit with respect to plant height and fresh top weight of celery to M. incognita race 1 was estimated as 0·15 eggs and J₂ mL^?1 soil. The minimum relative values (m) for plant height and top fresh weight were 0·37 and 0·35, respectively, at Pi ≥ 16 eggs and J₂ mL^?1 soil. The maximum nematode reproduction rate (Pf/Pi) was 407·6 at an initial population density (Pi) of 4 eggs and J₂ mL^?1 soil.     

Pathogenicity of the root-knot nematode Meloidogyne javanica on potato

Host–parasite relationships and pathogenicity of Meloidogyne javanica on potatoes (newly recorded from Malta) were studied under glasshouse and natural conditions. Potato cvs Cara and Spunta showed a typical susceptible reaction to M. javanica under natural and artificial infections, respectively. In potato tubers, M. javanica induced feeding sites that consisted of three to four hypertrophied giant cells per adult female. Infection of feeder roots by the nematode resulted in mature large galls which usually contained at least one mature female and egg mass. In both tubers and roots, feeding sites were characterized by giant cells containing granular cytoplasm and many hypertrophied nuclei. Cytoplasm in giant cells was aggregated alongside the thickened cell walls. Stelar tissues within galls appeared disorganized. The relationship between initial nematode population density ( P ) [0–64 eggs + second-stage juveniles (J2s) per cm³ soil] and growth of cv. Spunta potato seedlings was tested under glasshouse conditions. A Seinhorst model [ y = m  + (1 −  m ) z ^{( P − T )}] was fitted to fresh shoot weight and shoot height data of nematode-inoculated and control plants. Tolerance limits ( T ) for fresh shoot weight and shoot height of cv. Spunta plants infected with M. javanica were 0·50 and 0·64 eggs + J2s per cm³ soil, respectively. The m parameter in that model (i.e. the minimum possible y -values) for fresh shoot weight and shoot height were 0·60 and 0·20, respectively, at P  = 64 eggs + J2s per cm³ soil. Root galling was proportional to the initial nematode population density. Maximum nematode reproduction rate was 51·2 at a moderate initial population density ( P  = 4 eggs + J2s per cm³ soil).     

Solarization of soil in piles for the control of Meloidogyne incognita in olive nurseries in southern Spain

The potential of solarization to control Meloidogyne incognita in piles of soil used at olive nurseries in southern Spain was studied in 1999 and 2000. Kaolin and soil infested with free eggs and egg masses of the nematode in nylon bags were buried 20 and 40 cm deep inside conical piles of soil 80 cm high and with a base diameter of 1 m. Soil piles were solarized for 3 weeks in July and August. The effect of various periods of solarization was assessed by egg hatch bioassays in sterile water, and by infectivity to tomato plants. Maximum soil temperature at 20 cm depth in solarized piles was 47·4°C in 1999 and 48·2°C in 2000, compared with 32·9°C and 31·7°C in nonsolarized piles. Solarization reduced egg hatch by > 95% compared with nonsolarized samples, irrespective of type, burial depth and location of inocula in a soil pile. Egg hatch of egg mass-infested samples buried at 20 cm depth was higher than that of free eggs buried at the same depth. The differential effect associated with burial depth and type of inoculum was not found in solarized piles. In nonsolarized piles, hatch of free eggs from samples buried at 40 cm depth was higher than that from samples buried at 20 cm depth. Egg hatch in samples from solarized piles was lower than that from nonsolarized piles. A bioassay of tomato plants in 2000 confirmed the reduction in infectivity of free eggs buried in solarized soil piles. Under the conditions in southern Spain, solarization of 40 cm-high piles of soil for 3 weeks can therefore be used for the control of root-knot nematodes in potting soil for olive nursery production.     

Host suitability and response of different vegetable genotypes to Meloidogyne incognita race 2 and Meloidogyne javanica in South Africa

The host suitability of 20 locally available genotypes of Capsicum, 10 of Daucus carota, 7 of Beta vulgaris- and 3 of Spinacea oleracea were assessed in separate greenhouse studies for resistance to Meloidogyne incognita race 2 and M. javanica, respectively. Substantial variation existed amongst the vegetable genotypes in the greenhouse screenings with regard to their host status to the respective root-knot nematode species. None of the genotypes of D. carota, B. vulgaris and S. oleracea showed resistance to the nematode species tested. However, resistance to M. incognita race 2 was identified in Capsicum genotype “Tobasco”, which was subsequently verified in a follow-up microplot trial using a range of initial population densities together with a susceptible Capsicum genotype “Paprika”. Reproduction factors of the nematodes were used as the main criterion to evaluate resistance. In the microplot trial, genotype “Tobasco” showed resistance at the lower inoculum levels but not at the higher nematode population levels. The need exists for more frequent and extensive screenings of the various vegetable genotypes in order to provide small-scale farmers with better options for improved and sustainable yields.     

粉红螺旋聚孢霉NF-06固体发酵条件优化及对南方根结线虫的防治效果

南方根结线虫是一种严重危害我国蔬菜安全生产的土传病害,生产上亟需一种有效的防控方法。本研究从根结线虫卵囊中分离到一株生防真菌NF-06,通过形态学和分子生物学方法将其鉴定为粉红螺旋聚孢霉Clonostachys rosea。通过离体活性测定的方法测定NF-06发酵滤液对根结线虫2龄幼虫的致死作用,发现24、48和72 h后2龄幼虫的相对死亡率分别达63.6%,75.5%和87.0%。以产孢量为指标,优化菌株NF-06固体发酵条件,确定最佳基础培养基为麦麸和麦秸秆混合物,其比例为4：1（wt/wt）,在基础培养基中添加10%玉米粉、2%蚕豆粉和0.05% KH₂PO₄可获得最大产孢量;最优培养条件为初始pH值7.0,初始含水量40%,接种量5%,产孢量达8.9×10¹⁰ CFU/g。室内盆栽防治效果测定显示,菌株NF-06固体发酵物能够有效降低土壤中的线虫数量和番茄根结指数,其中5 g/株穴施处理的线虫减退率和防治效果分别达67.9%和69.6%。本研究为生物杀线虫剂的开发和利用提供理论依据。     

Host suitability of Solanum torvum cultivars to Meloidogyne incognita and M. javanica and population dynamics

Several experiments were carried out to assess the performance of commercial Solanum torvum cultivars against the root knot nematodes Meloidogyne incognita and M. javanica in Spain. The response of S. torvum rootstock cultivars Brutus, Espina, Salutamu and Torpedo against M. incognita and Mi-1.2 (a)virulent M. javanica isolates was determined in pot experiments, and of ‘Brutus’ to an N-virulent isolate of M. incognita, compared with that of the eggplant S. melongena ‘Cristal’. The relationship between the initial and final population densities of M. javanica on ungrafted and grafted ‘Cristal’ onto the S. torvum ‘Brutus’ was assessed, together with the effect on dry shoot biomass. Finally, the population growth rate and the resistance level of the four S. torvum cultivars against M. incognita was assessed under plastic greenhouse conditions in two cropping seasons. All S. torvum rootstocks responded as resistant to the M. incognita isolates and from highly resistant to susceptible against M. javanica isolates. The maximum multiplication rates of M. javanica on the ungrafted or grafted eggplant were 270 and 49, respectively, and the equilibrium densities were 1318 and 2056 eggs and J2 per 100 cm³ soil, respectively. The tolerance of the ungrafted eggplant was 10.9 J2 per 100 cm³ soil, and the minimum relative dry shoot biomass was 0.76. The population growth rate of M. incognita on eggplant cv. Cristal differed from that of the S. torvum cultivars in both cropping seasons. These results suggest that S. torvum is a valuable rootstock for managing the two Meloidogyne species irrespective of the (a)virulence status.     

Pathozone dynamics of Meloidogyne incognita in the rhizosphere of tomato plants in the presence and absence of the nematophagous fungus, Pochonia chlamydosporia

Pathozone dynamics were derived for approx. 50 Meloidogyne incognita juveniles infecting a single root of tomato under controlled conditions from a point source of inoculum and described using simple, non-linear models. The pathozone decayed sigmoidally with distance, but increased over time as progressively more nematodes were able to infect the root. Despite the reported ability of M. incognita juveniles to travel up to 50 cm in some conditions, the maximum width of the pathozone for a single tomato root was estimated as 18·1 mm. It is conjectured that this was because of (i) diffusion from a point source of inoculum, (ii) a small infection court (a single root tip) and (iii) the limited life span of the nematode. A second experiment was used to assess the effect of the nematophagous fungus Pochonia chlamydosporia on the pathozone dynamics of the nematode. This fungus is known to produce nematicidal products in vitro , which may affect invasion of roots by the free-living nematode. To examine the possibility of a change in the position of the site of infection, changes in the probability of gall formation along the root length were also examined. In the absence of P. chlamydosporia , the pathozone dynamics of M. incognita were very similar to those of the first experiment. It was shown that P. chlamydosporia did not significantly affect the pathozone dynamics of M. incognita nor the site of gall formation, which appear to have little importance for the role of the fungus as a biological control agent.     

Screening of chemical attractants for second-stage juveniles of Meloidogyne species on agar plates

The attractiveness of 60 aromatic compounds, mainly carboxylic acids, alcohols, aldehydes, and phenols, to second-stage juveniles (J2) of four Meloidogyne species (M. hapla, M. incognita, M. javanica, and M. marylandi) was evaluated based on the relative density of J2 attracted to a test compound on an agar plate in an 8.5-cm Petri dish. Three types of nematode responses were observed in the single-compound assays: attraction, in which J2 were attracted to test compounds; concentration-dependent attraction, in which J2 were attracted to the zone located 1–2 cm from test compounds; and no response. None of the test compounds effectively attracted M. incognita or repelled J2 of any species. Thirty-five compounds attracted J2 of at least one of the other three nematode species. Highly effective attractants were trans-cinnamic acid, salicylic acid, 4′-hydroxy-3′-methoxyacetophenone, o-vanillin, carvacrol, 3-methoxybenzyl alcohol, 2-methoxycinnamaldehyde, trans-p-methoxycinnamaldehyde, 4-methoxy-3-methylbenzaldehyde, 2-methoxy-4-propenylphenol, and thymol. Salicylic acid dissolved in the agar reduced the attraction of M. marylandi J2 to salicylic acid and to o-vanillin. Two- and three-compound assays revealed differences in attractiveness to M. marylandi J2 between or among structural isomers of the attractants. o-Vanillin and salicylic acid were much more attractive to M. marylandi than vanillin and the two isomers of salicylic acid, respectively. It is not clear whether nematodes utilize the attractants found in the study for locating hosts in nature, but the attractants may have potential for use in nematode control.     

Weed hosts of Meloidogyne spp. and the effect of aqueous weed extracts on egg hatching

Weeds have a great economic impact on agricultural production because they compete with crops for resources and are alternative hosts for pests, microbial pathogens and plant-parasitic nematodes. This study aimed to investigate the susceptibility of weeds to the root-knot nematodes Meloidogyne javanica and Meloidogyne incognita and assess the effect of aqueous weed extracts on M. javanica egg hatching. Four experiments were conducted, two for each nematode species. Weeds were inoculated with 2000 nematode eggs and grown for 60 days under greenhouse conditions. Soyabean cv. Monsoy 7110 was used as control. The weeds Ipomoea grandifolia, Solanum americanum, Digitaria horizontalis, Amaranthus deflexus, Sorghum halepense and Commelina benghalensis were susceptible to M. javanica and M. incognita in at least one experiment (reproduction factor, RF >1). Crotalaria juncea and Eleusine indica were susceptible to M. incognita in one experiment, and Digitaria insularis, Sida rhombifolia, Bidens pilosa, Urochloa decumbens, Crotalaria breviflora, Cenchrus echinatus, Crotalaria ochroleuca and Crotalaria spectabilis were immune or resistant (RF <1 or RF = 0). Alternanthera tenella, C. juncea, S. rhombifolia, C. ochroleuca, C. spectabilis, C. breviflora, B. pilosa, E. indica, U. decumbens and C. echinatus were resistant or immune to M. javanica (RF <1 or RF = 0). Compared with the control (water), all weed extracts reduced M. javanica egg hatch. Our results highlight the importance of weed control in agricultural systems, as nematodes can survive and multiply in weed roots during the off-season. Weed leaves and shoots, however, may be an interesting source of compounds with nematicidal activity.     

Fitness cost but no selection for virulence in Meloidogyne incognita after two consecutive crops of eggplant grafted onto Solanum torvum

The eggplant Solanum melongena cv. Cristal, either ungrafted or grafted onto the Solanum torvum rootstock cv. Brutus, was cultivated for two consecutive years in the same plots in a plastic greenhouse to assess the level of resistance to Meloidogyne incognita and crop yield. At the end of the second crop, the putative selection for virulence of the nematode subpopulations coming from infected ungrafted and grafted eggplant was assessed in the eggplant and in S. torvum in a pot experiment. Nematode population densities at transplantation in 2017 ranged from 2 to 378 per 100 cm³ of soil and did not differ between ungrafted and grafted eggplant. At the end of each crop, a higher galling index and number of nematodes in soil and in roots was registered in ungrafted compared to grafted eggplant. The grafted eggplant was categorized as resistant in 2017 and as highly resistant in 2018. Eggplant yield did not differ irrespective of grafting in 2017 after being cultivated for 135 days, but it differed after 251 days of cultivation in 2018. In the pot experiment, S. torvum was categorized as resistant to both M. incognita subpopulations. However, the M. incognita subpopulation obtained from roots of S. torvum produced 49.4% fewer egg masses and 56% fewer eggs per plant in the eggplant than the nematode subpopulation obtained from roots of the eggplant cv. Cristal. The results of this study revealed that the infective and reproductive fitness of the nematode decreased without having been selected for virulence.     

Selection of virulent populations of Meloidogyne javanica by repeated cultivation of Mi resistance gene tomato rootstocks under field conditions

Field trials were conducted in a plastic house artificially infested with an avirulent population of Meloidogyne javanica to determine the durability of the resistance mediated by the Mi gene in tomato rootstocks after repeated cultivation for three consecutive years. Treatments included an experimental rootstock cv. PG76 ( Solanum lycopersicum  ×  Solanum sp.), a commercial rootstock cv. Brigeor ( S. lycopersicum  ×  S. habrochaites ), a resistant tomato cv. Monika ( S. lycopersicum ), and a susceptible cv. Durinta ( S. lycopersicum ). Based on the reproduction index (RI: number of eggs per g root on the resistant cultivar divided by number of eggs per g root on the susceptible cultivar × 100), rootstock cv. PG76 responded as highly resistant (RI = 7%) after the first cropping cycle (3·4 nematode generations), showed intermediated resistance (RI = 33%) after the second cropping cycle (3·3 generations), and was fully susceptible (RI = 94%) after the third cycle (3·3 generations). In contrast, rootstock cv. Brigeor and resistant cv. Monika retained intermediate resistance levels (RI = 41 and 25%, respectively) after the third cropping cycle. Virulent nematode populations were rapidly selected from an avirulent one after repeated cultivation of resistant tomatoes under field conditions. Bioassays conducted under controlled conditions confirmed that selection for virulence occurred more rapidly in plots with cv. PG76 followed by Brigeor and Monika. The nematode population in the field not exposed to Mi resistance remained avirulent to Mi genotypes. The genetic background of the resistant rootstocks and the frequency of cropping were critical factors for the appearance of virulent nematode populations. Irrespective of nematode infection, all resistant tomatoes yielded more than the susceptible cultivar.     

Systemic nematicidal activity of fluensulfone against the root-knot nematode Meloidogyne incognita on pepper

BACKGROUND: Fluensulfone, a new nematicide of the fluoroalkenyl group, has proved to be very effective in controlling root‐knot nematodes, Meloidogyne spp., by soil application. The systemic activity of this compound against M. incognita on peppers via soil drenching and foliar spray was evaluated. RESULTS: Root application of fluensulfone via soil drenching showed slight and no nematode control activity when applied 4 and 10 days, respectively, after inoculation. A single foliar spray of peppers with a fluensulfone solution at 3.0 g L^?1 prior to inoculation reduced the galling index by 80% and the number of nematode eggs by 73–82% of controls. The reduction in these parameters by fluensulfone was much higher than that obtained with oxamyl or fenamiphos at the same concentration. This activity was also observed when the plants were sprayed 21 days before inoculation. A series of experiments suggested that foliar spray with fluensulfone prior to inoculation reduces nematode invasion. However, foliar spray after inoculation did not inhibit nematode development inside roots. CONCLUSION: Fluensulfone showed relatively high nematode control activity when sprayed on the foliage before inoculation. Fluensulfone may be used as a foliar application, in addition to soil application, for root‐knot nematode control. Copyright © 2011 Society of Chemical Industry     

植物源提取物防治根结线虫的分子研究

针对内生真菌的生物防治,重点阐述了中草药提取物的杀虫活性、植物抗线虫基因的发现以及植物防御系统激发的最新研究.植物源提取物作为潜在的线虫杀虫剂,为生物源农药开发和利用提供新的思路.同时,对于今后线虫杀虫剂开发中存在的问题及解决方案进行了探讨,就其发展方向提出了建议.     

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.     

植物根结线虫基因组学研究进展

根结线虫是世界农业生产中危害最大的植物病原之一,目前仍缺乏安全有效的防治措施。深入揭示寄生线虫与植物之间互作的分子机制,利用生物技术进行抗性育种被认为是最有前景的抗线虫策略。在根结线虫基因组学研究方面,目前已经构建了北方根结线虫AFLP遗传连锁图谱,南方根结线虫和北方根结线虫基因组测序也已完成;基因组的注释和比较基因组学分析,较全面地描述了根结线虫的遗传组成;以差异表达分析和比较基因组学为主的方法鉴定了大量的重要基因;以RNA干扰、植物转化和蛋白互作为主的根结线虫基因功能研究也取得了一些进展。本文就根结线虫基因组学研究予以综述,并进一步探讨其研究方向和可持续抗线虫新策略的发展前景。     

Molecular diagnostic key for identification of single juveniles of seven common and economically important species of root-knot nematode (Meloidogyne spp.)

A molecular protocol is presented for distinguishing seven of the most common and economically important Meloidogyne spp. DNA was extracted from individual second-stage juvenile (J2) nematodes of Meloidogyne spp. and amplified by PCR (polymerase chain reaction). Fifteen PCRs including amplification of rDNA, specific SCAR (sequence characterized amplified region) and RAPD (random amplified polymorphic DNA) fragments were possible from the extracted DNA. This enabled a molecular diagnostic key for M. incognita , M. javanica , M. arenaria , M. mayaguensis , M. hapla , M. chitwoodi and M. fallax to be designed. The key unifies published methods into a single logical schematic using primer combinations that were previously validated and shown to work reliably and specifically. The protocol can be used with single juvenile or adult nematodes and the schematic can readily be expanded to accommodate more species. The use of RAPD amplification to assist with identification of samples which do not yield diagnostic amplification products after the first three steps of the molecular key is also described.