Genetic and Phenotypic Diversity in the Graminaceous Cyst Nematode Complex, Inferred from PCR-RFLP of Ribosomal DNA and Morphometric Analysis

Graminaceous cyst nematodes form a group of eleven valid species including Heterodera avenae, Heterodera filipjevi and Heterodera latipons and constitute major pests to cereals. They are widely spread in circum-mediterranean areas where they are presumed to cause yield losses on bread and durum wheat. The objective was to document the diversity of these cereal cyst nematodes, in particular samples from Mediterranean regions, in comparison to species which develop on cultivated or wild grasses (H. arenaria, H. hordecalis, H. mani) and on rice or sugarcane (H. sacchari). Studies involved PCR-RFLP of ITS and morphometrics of the juvenile and cyst characters. UPGMA analysis of molecular data showed that the isolates segregated in two main clusters which seem to represent different evolutionary lineages. The H. avenae sensu lato cluster (I) contained H. arenaria, H. avenae, H. filipjevi and H. mani. The second cluster (II) contained isolates of H. hordecalis and H. latipons. Within H. avenae sensu lato, H. filipjevi was separated from the other related species with significant bootstrap value. The differentiation of H. arenaria, recognized for the first time based on molecular data, and H. mani with few restriction enzymes were the least significant. Intraspecific polymorphism allowed differentiation of isolates originating from Australia within H. avenae sensu stricto. The group H. hordecalis–H. latipons showed the greatest genetic variability between and within isolates. Two representatives of Heterodera sacchari, taxonomically included in the schachtiigrave group, were genetically as distant to this group as to the other graminaceous species belonging to either H. avenae sensu lato or H. hordecalis–H. latipons group. Results inferred from multivariate analysis applied on morphometrics of the cysts and juveniles showed agreement between genetic and phenotypic classifications. This study demonstrates the utility of combined molecular and classical methods to enhance our knowledge about the diversity within the complex of graminaceous cyst nematodes and to establish robust techniques to identify a wider set of nematode species.     

Influence of different fertilization regimes on plant-parasitic nematodes in the black soil region of Northeast China

Plant-parasitic nematodes can cause serious plant diseases and adversely affect crop production. We investigated the temporal and vertical dynamics of plant-parasitic nematodes in northeast China to determine if long-term fertilizer application suppresses the population of plant-parasitic nematodes and influences their vertical distribution in the black soil region. The long-term fertilizer application lasted for 14 years, and included three treatments: pig manure combined with chemical fertilizer (MCF), chemical fertilizer (urea and ammonium phosphate, CF), and no fertilizer (NF). A 3-year corn-wheat-soybean rotation was grown on the field site, soil samples were obtained in the corn phase and nematodes were extracted, identified and counted. Six plant-parasitic nematode genera with relative abundance over 0.1% were found in this study. Heterodera was the dominant genus in all three fertilizer treatments, and its relative abundance was highest in NF (45.4%) and lowest in MCF (32.8%). Fertilizer application had a significant effect on abundance of total soil nematodes but not on either plant-parasitic nematodes or Heterodera. Significant differences in the abundance of total soil nematodes, plant-parasitic nematodes and Heterodera were observed among soil depths and interaction of fertilizer × soil depth. Total soil nematodes were mainly distributed in the 0–20 cm soil layer, and plant-parasitic nematodes and Heterodera were mostly distributed in the 10–30 cm soil layer. Fertilizer application did not show a significant effect on plant-parasitic nematodes or Heterodera at any of the sampling depths from 0 to 80 cm. The results indicated that fertilizer has no effective control on plant-parasitic nematodes when dominant genera and their respective plant hosts exist in relatively high abundance.     

河南省禾谷类作物孢囊线虫的发生分布与种类鉴定

为探究河南省主要禾谷类作物孢囊线虫的发生分布,明确孢囊线虫对不同作物的危害情况,于2017—2020年对河南省18个市50个县（区）的小麦、玉米和水稻作物的孢囊线虫种类和发生分布进行随机取样调查,采用形态学特征、分子生物学鉴定和rDNA-ITS序列进化树分析技术鉴定不同作物孢囊线虫的种类,并根据土壤中孢囊基数和单孢囊卵数明确不同作物孢囊线虫的发病严重度。结果显示,共采集土壤样品308份,其中224份样品检测到孢囊,孢囊检出率为72.7%。小麦孢囊线虫发生分布范围覆盖40个县（区）,其中15个县（区）为禾谷孢囊线虫Heterodera avenae侵染,23个县（区）为菲利普孢囊线虫H. filipjevi侵染,南阳市西峡县西坪镇和开封市尉氏县张市镇为禾谷孢囊线虫和菲利普孢囊线虫混合侵染发生区;玉米孢囊线虫H. zeae在濮阳市清丰县韩村镇、许昌市长葛市董村镇和禹州市范坡镇检测点首次被发现;旱稻孢囊线虫H. elachista在信阳市潢川县魏岗乡、来龙乡和新乡市获嘉县亢村镇检测点首次被发现。孢囊线虫发病严重度数据表明,小麦田平均孢囊含量高达17.3个/100 mL;玉米田平均孢囊量为11.0个/100 mL;水稻田平均孢囊量为4.4个/100 mL。表明河南省孢囊线虫高发地块主要集中在豫北、豫东和豫中平原区。     

The incredible fungal genus —Drechslera — and its phytotoxic ophiobolins

ManyDrechslera species that haveCochliobolus sp. as a perfect stage produce a related group of ophiobolins, sesterterpenoids (C-25’s), which are phytotoxic to a wide range of plants. One of the most toxic ophiobolins is 6-epiophiobolin A and it is produced by all of theDrechslera species thus far studied. On the other hand, some novel ophiobolins have been found inD. oryzae: ophiobolin J, 6-epiophiobolin I, and 8-deoxyophiobolin J. Potentially, genetic crosses made between these fungi could yield novel organisms, producing novel combinations of the ophiobolins, and thus new pathotypes. Ophiobolins are being used in tissue culture systems to screen plants for sensitivity to these toxins. Paper presented at the Bat-Sheva Seminar on Host - Fungus Interaction, Jerusalem, Israel (March 14–25,1988).     

Interaction of Root-knot Nematodes (RKN) and the Bacterium Agrobacterium Tumefaciens in Roots of Prunus Cerasifera: Evidence of the Protective Effect of the Ma RKN Resistance Genes Against Expression of Crown Gall Symptoms

Agrobacterium tumefaciens (AT) is the causal agent of crown gall, a major problem in the family Rosaceae and particularly for Prunus spp. Crown gall symptoms result from the bacterial infection of the cells damaged mechanically at the collar or by root parasitic nematodes. Myrobalan plum (P. cerasifera) is susceptible to AT and is not a host for the root-knot nematode (RKN), M. hapla. Some clones of this plum carry single Ma resistance genes that control M. arenaria, M. incognita and M. javanica. The four above mentioned RKN and Myrobalan progenies segregating for Ma were used in experiments aimed at obtaining a better knowledge of the interaction between AT and RKN in relation to the RKN resistance genes. Prunus rooted cuttings, naturally infected with the bacterium were repotted, grown and inoculated individually with RKN. In a first experiment, Prunus plants were (i) either inoculated with 10,000 juveniles (J2s) of M. arenaria to provide a short inoculum pressure (SIP) or (ii) inoculated by association with one M. arenaria-galled tomato root system that produced a high and durable inoculum pressure of the same nematode species. Four months after RKN inoculation, plants were rated for nematode and bacterial root galling symptoms. RKN and AT galls were more numerous and more homogenous under DIP than under SIP. Nevertheless, for both inoculum regimes, AT galls were present in the RKN-susceptible clones (= carrying none of the Ma genes) and absent in the RKN-resistant clones. Subsequent experiments, conducted under DIP with M. arenaria, M. incognita, M. javanica and M. hapla, also showed, for the three first species, the presence of AT galls only in RKN-susceptible clones whereas Prunus plants inoculated with M. hapla and nematode-free controls were free of AT galls. Consequently RKN act as a wound agent in the AT infection process of Myrobalan plum only when the plant develops a compatible reaction (i.e. when it lacks the Ma resistance genes). Considering that J2s do penetrate the roots of resistant plants, the absence of crown gall symptoms on this material even under durable inoculum pressure strengthens the hypothesis that this nematode stage has a very weak effect on plant cells during the infection process. This is the first evidence of the protective effect of a RKN resistance gene against the expression of root crown gall consecutive to RKN infection. The protective effect of Ma and presumably of other RKN resistance genes against AT is a strong argument for their introgression into Prunus and other Rosaceae or bacterium-susceptible crops.     

A fungistatic chromene from<Emphasis Type="Italic">Ageratum conyzoides</Emphasis>

Ageratum conyzoides L. is an annual herb in the tropics and subtropics whose extracts are known to possess pharmacological and biocidal activity. We report on the bioactivity of a secondary metabolite (a chromene) isolated from the shoots ofA. conyzoides against some plant pathogenic fungi. Organic solvent extracts from the shoots were tested for antifungal activity against the plant pathogenic fungiRhizoctonia solani, Sclerotium rolfsii, Botryodiplodia theobromae, Phomopsis theae andFusarium species growingin vitro on potato dextrose agar medium. The cruden-hexane extract completely inhibited the growth ofR. solani andS. rolfsii. Then-hexane extract was chromatographed over a column of silica gel followed by activity-guided fractionation to give an antifungal principle. Structure elucidation by detailed analysis of¹H,¹³C NMR and mass spectroscopy identified the active compound as precocene II. The growth ofR. solani andS. rolfsii was completely inhibited by precocene II at a concentration of 80–100 ppm. The sclerotia ofR. solani andS. rolfsii were also completely suppressed by 150 ppm of precocene II. Sub-culture of these inhibited fungi onto precocene II-free medium restored growth of the fungus, indicating that precocene II is fungistatic. Crude or refined extracts fromA. conyzoides offer the possibility of biocontrol of plant pathogenic fungi. http://www.phytoparasitica.org posting Feb. 11, 2004.     

Note: Extrapolating the use of an entomopathogenic nematode and fungus as control agents forFrankliniella occidentalis toThrips palmi

Little is known regarding non-chemical control measures againstThrips palmi Karny. Since entomopathogenic fungi and nematodes have been found to be active against thrips species such asFrankliniella occidentalis Pergande, comparative bioassays were conducted to determine the extent to which they also show activity againstT. palmi. Significant mortality of the larvae of the species was recorded following treatment withLecanicillium muscarium (Petch) Zare and Gams, and addition of the wetting agent Agral enhanced pathogenicity toT. palmi. T. palmi pupae were not affected byS. feltiae. The potential for use of these agents againstT. palmi in the field is discussed in the light of these results. http://www.phytoparasitica.org posting Aug. 28, 2005.     

Toxicity of fungal endophyte secondary metabolites to plant parasitic nematodes and soil-borne plant pathogenic fungi

Fungi isolated from the cortical tissue of surface sterilized tomato roots collected from field plots produced secondary metabolites in nutrition broth that were highly toxic toMeloidogyne incognita. Especially strains ofFusarium oxysporum were highly active with 13 of 15 strains producing culture filtrates toxic to nematodes. The mechanism of action of the toxic metabolites produced by the non-pathogenicF. oxysporum strain 162 with proven biological control ofM. incognita in pot experiments was investigated. These metabolites reducedM. incognita mobility within 10 min of exposure. After 60 min, 98% of juveniles were inactivated. Juveniles were initially inactivated within a few minutes of exposure, but with exposure of 5 h 50% of the juveniles were dead and 24 h exposure resulted in 100% mortality. In a bioassay with lettuce seedlings metabolite concentrations > 100 mg/l reduced the number ofM. incognita juveniles on the roots comparing to the water control. TheF. oxysporum toxins were highly effective towards sedentary parasites and less effective towards migratory endoparasites. Nonparasitic nematodes were not influenced at all. Metabolites of strain 162 also reduced significantly the growth ofPhytophthora cactorum, Pythium ultimum andRhizoctonia solani in vitro. Secondary metabolites of endophytic fungi on plant-parasitic nematodes and soil-borne fungi should be considered for control of plant parasitic nematodes and plant pathogenic fungi. The results also show the need for proper selection of target nematodes inin vitro bioassays.     

Note: Molecular identification of three entomopathogenic nematodes from turkey by PCR-RFLP of the ITS regions

Molecular identification methods are widely used for the classification of organisms worldwide. Entomopathogenic nematodes are the most often isolated insect parasitic nematodes in the tropical and subtropical regions. In our investigation, PCR-RFLP (Polymerase Chain Reaction — Restriction Fragment Length Polymorphism) of the ITS region (Internal Transcribed Spacer) on the ribosomal (r) DNA of three entomopathogenic nematodes isolated from Ankara, Turkey, was analyzed for identification. The ITS region of rDNA was amplified by PCR and then digested with the following nine restriction enzymes: Alu I, Dde I, Hae III, Hha I, Hind III, Hinf I, Hpa II, Rsa I and Sau 3AI. The amplified and restricted sequences of the ITS regions were separated by agarose gel electrophoresis and the RFLP patterns of these three species were shown in this study. According to our results, these species were identified asSteinernema feltiae, Steinernema carpocapsae andHeterorhabditis bacteriophora. http://www.phytoparasitica.org posting Nov. 4, 2005.     

Evaluation of Natural Chemical Compounds Against Root-lesion and Root-knot Nematodes and Side-effects on the Infectivity of Arbuscular Mycorrhizal Fungi

The survival of two species of plant parasitic nematodes: the root-lesion nematode Pratylenchus brachyurus, and the root-knot nematode Meloidogyne javanica, was evaluated in saturated atmospheres of 12 natural chemical compounds. The infectivity of two isolates of arbuscular mycorrhizal fungi: Glomus mosseae and Glomus intraradices, under identical experimental conditions, was also determined. All the compounds tested exerted a highly significant control against M. javanica and among them, benzaldehyde, salicilaldehyde, borneol, p-anisaldehyde and cinnamaldehyde caused a mortality rate above 50% over P. brachyurus. The infectivity of G. intraradices was inhibited by cinnamaldehyde, salicilaldehyde, thymol, carvacrol, p-anisaldehyde, and benzaldehyde, while only cinnamaldehyde and thymol significantly inhibited mycorrhizal colonization by G. mosseae.     

Production of phytotoxic metabolites by five species of Botryosphaeriaceae causing decline on grapevines, with special interest in the species Neofusicoccum luteum and N. parvum

In recent years an increasing number of species of Botryosphaeriaceae have been associated with grapevine decline worldwide. Five species isolated from declining grapevines in Spain (Botryosphaeria dothidea, Diplodia seriata, Dothiorella viticola, Neofusicoccum luteum and N. parvum) were checked for toxin production in liquid cultures. Cultural conditions for all fungi were adjusted to obtain optimal production of phytotoxic culture filtrates, by growing the fungi in steady liquid cultures of Czapek–Dox broth for different time intervals. Phytotoxicity of D. seriata and N. parvum reached a maximum after 14 days while the remaining species showed the highest phytotoxicity levels after 21 days in culture. All fungi produced hydrophilic high-molecular weight compounds with phytotoxic properties. In addition, N. luteum and N. parvum produced lipophilic low-molecular weight phytotoxins, not detected consistently among the remaining species. This led to a more exhaustive study on the phytotoxicity of N. luteum and N. parvum. Culture filtrates and corresponding extracts of both species were consistently highly phytotoxic in different assays. The gas-chromatography analysis of the acetylated O-methyl glycosides of the phytotoxic exopolysaccharides produced by N. parvum showed these substances to be composed mainly of glucose, mannose and galactose. Results suggest that phytotoxic metabolites could be involved in the virulence of both species in planta.     

The use of Solacol® (validamycin) as a growth retardant in the isolation of soil fungi

Solacol^®, a formulation of the antibiotic validamycin, at 0.33% in 2% malt extract agar, reduced the spread of fungi on dilution plates drastically and allowed twice as much incubation time before subculturing; this resulted in an elevated number of species isolated. Using pure cultures of 62 common soil fungi, it was shown that all fast-growing species (exceptPythium ultimum) were efficiently inhibited but not completely suppressed. Inhibition was comparable to that by 0.5% oxgall, though, while this substance completely suppressed several species, Solacol very strongly inhibited onlyGaeumannomyces graminis, Gerlachia nivalis, Harzia acremonioides, Verticillium biguttatum andRhizoctonia solani. In a further experiment each separate constituent of Solacol was tested against 22 fungi at equivalent concentrations. Validamycin strongly inhibitedChaetomium globosum and two Basidiomycetes, though hardly more than the non-ionic detergent which mainly inhibited the other fungi. A few species were, however, more inhibited by Solacol than by the detergent alone. Solacol at 0.33% is a suitable aid in dilution plating of soil fungi, by increasing the number of colonies and species observed.Samenvatting Solacol^®, een formulering van het antibioticum validamycine, remde de groei van schimmels in verdunningsplaten met een concentratie van 0.33% in 2% moutagar en maakte het mogelijk de periode tot afenten met een factor 2 te verlengen; daardoor was het aantal geïsoleerde soorten duidelijk toegenomen. Met reincultures van 62 algemene grondschimmelsoorten werd aangetoond, dat alle snelgroeiende soorten (met uitzondering vanPythium ultimum) voldoende geremd, maar niet volkomen onderdrukt werden. Het remmingspercentage was vergelijkbaar met dat van 0.5% ossegal, hoewel dit laatste sommige soorten volkomen onderdrukte; Solacol remde alleenGaeumannomyces graminis, Gerlachia nivalis, Harzia acremonioides, Verticillium biguttatum enRhizoctonia solani zeer sterk. In een volgend experiment werden de componenten van Solacol t.o.v. 22 fungi apart getoetst in concentraties equivalent aan 0.33% Solacol. Validamycine remde alleenChaetomium globosum en twee basidiomyceten behoorlijk, maar nauwelijks meer dan de niet-ionische uitvloeier, die in hoofdzaak de overige remeffecten veroorzaakte. Enkele soorten werden echter door het complete Solacol veel sterker geremd dan door de uitvloeier alleen. Solacol in een verdunning van 0,33% wordt aanbevolen bij verdunningsplaten voor het isoleren van grondschimmels ten einde het aantal kolonies en soorten te verhogen.     

The influence of some agricultural practices on soil organisms and plant establishment of sugar beet

In a long term field experiment the influence of crop rotation and the application of insecticides and herbicides on soil arthropods (mesofauna), nematodes, soil fungi and weeds was investigated. Possible changes in numbers of not only pathogenic organisms, but also of predators or parasites of pathogens, could affect seedling establishment and/or yield of sugar beet. In soil samples taken during the spring, high population of the springtailOnychiurus armatus ware detected which caused considerable damage in the first two years. Later also rather severe infestations byAtomaria linearis occurred, probably caused by frequent sugar beet cropping.No significant influence of crop rotation on damage caused by soil arthropods could be detected yet, but in the 1/3 sugar beet and 2/3 winter wheat rotation more micro-arthropods and a higher diversity of species were found than in the blocks with continuous sugar beet growing. In the latter ones the numbers ofHeterodera schachtii increased and in 1980 the tolerance level was exceeded. On the contrary in the 1/3 sugar beet blocksPratylenchus spp. were dominant. Since a number of perennial weeds cannot be controlled sufficiently in sugar beet, some of them created a problem in the 1/1 sugar beet blocks.In the plots not treated with a soil herbicide large numbers of annual weeds provided alternative native food forO. armatus, which resulted in less plant damage. No significant effect of chloridazon on the micro-arthropod fauna could be detected. The insecticides lindane and aldicarb controlledO. armatus andA. linearis partially, and improved seedling emergence and to a lesser extent yield. Both insecticides had a variable effect on the relative abundancy ofCollembola and in addition lindane suppressed numbers ofAcariformes and aldicarb those ofParasitiformes. No negative influence on the species diversity could be detected. All effects did not last long and varied considerably from year to year.Indications were found that, if species ofCollembola, which could act as predators of nematodes, were suppressed, the numbers ofPratylenchus spp. andH. schachtii increased.Samenvatting In een veeljarig proefveld (1976–1979) werd de invloed van de vruchtopvolging en toepassing van insecticiden en herbiciden op bodemarthropoden (mesofauna), nematoden, bodemschimmels en onkruiden onderzocht. Niet alleen mogelijke veranderingen in de aantallen schadeverwekkers, maar ook in de aantallen predatoren en pathogenen van bodemplagen zouden het plantbestand en/of de opbrengst van suikerbieten kunnen beïnvloeden.Gedurende de eerste twee jaar van de proef werden in grondmonsters, die gedurende het voorjaar werden genomen, grote aantallen springstaarten (Onychiurus armatus) aangetroffen, die aanzienlijke schade veroorzaakten. Later kwamen eveneens aantastingen door het bietekevertje (Atomaria linearis) voor, waardoor in meerdere of mindere mate planten wegvielen.Er kon nog geen duidelijke invloed van de vruchtopvolging op de schade door bodemarthropoden worden vastgesteld, maar in de rotatie met 1/3 suikerbieten en 2/3 wintertarwe werden meer micro-arthropoden en een grotere soortenrijkdom gevonden dan in de gedeelten met een continuteelt van suikerbieten. In laatstgenoemde veldjes zijn de aantallen bietecystenaaltjes (Heterodera schachtii) toegenomen, waarbij in 1980 de tolerantiegrens werd overschreden. Daarentegen vond in de veldjes met 1/3 suikerbieten in de rotatie een sterkere vermeerdering vanPratylenchus spp. plaats dan in de continuteelt. Aangezien een aantal wortelonkruiden in suikerbieten niet voldoende kan worden bestreden, veroorzaakten enkele na enige jaren een probleem in de continue bietenteelt.In de veldjes die niet met een bodemherbicide waren behandeld vormden grote aantallen zaadonkruiden een alternatieve voedselbron voorOnychiurus armatus, wat een geringere aantasting van kiemplanten tot gevolg had. Er werd geen aantoonbaar effect van chloridazon op de micro-arthropoden vastgesteld.De insecticiden lindaan en aldicarb konden schade doorOnychiurus armatus enAtomaria linearis slechts gedeeltelijk voorkomen; er werd een duidelijk betere opkomst en soms ook een hogere opbrengst verkregen. Beide insecticiden hadden een wisselende invloed op de relatieve talrijkheid vanCollembola en bovendien onderdrukte lindaan de aantallenAcariformes en aldicarb deParasitiformes. Er kon geen negatieve invloed op de soortenrijkdom worden vastgesteld. Al deze effecten waren niet van langdurige aard en verschilden sterk van jaar to jaar.Er werden aanwijzingen gevonden dat, indien bepaalde soortenCollembola, die als predatoren van nematoden kunnen optreden, werden onderdrukt, de aantallenPratylenchus spp. enHeterodera schachtii toenamen.     

Occurrence and distribution of <Emphasis Type="Italic">Phytophthora</Emphasis> species in European chestnut stands,and their association with Ink Disease and crown decline

The Phytophthora complex associated with Castanea sativa Mill. was investigated in five European countries in 35 regions and with respect to various domestication levels. Annual precipitation and length of drought season were the main parameters that regulated the presence of Phytophthora species in the chestnut stands. Seven species of Phytophthora were detected; three of these, P. megasperma, P. cryptogea and P. syringae had not been previously reported on sweet chestnut. P. cinnamomi. P. cambivora and P. citricola were most frequently isolated. P. cinnamomi and P. cambivora were the species significantly associated with declining trees with symptoms of Ink Disease. P. cinnamomirequired distinct ecological conditions compared to the other species. P. cinnamomi was never detected in sites characterized by minimum temperatures below 1.4 °C, maximum temperature above 28 °C, or soil pH below 5.4. The results obtained provide useful information for modeling the probability of Ink Disease, crown decline and associated Phytophthora species in chestnut groves in global climatic change scenarios.     

Biological control potential of Turkish entomopathogenic nematodes against the Mediterranean fruit fly <Emphasis Type="Italic">Ceratitis capitata</Emphasis>

The entomopathogenic nematodes Steinernema weiseri, S. feltiae, S. carpocapsae and two strains of Heterorhabditis bacteriophora, isolated from Turkish soils, were evaluated against larvae of the Mediterranean fruit fly (medfly) Ceratitis capitata in plastic cups under laboratory conditions with sandy loam soil and 10% moisture level. At a rate of 100 infective juveniles (IJs)/cm², the last instar larvae of C. capitata were susceptible to the entomopathogenic nematodes: the S. feltiae 09-31 strain recovered from Aydin provided 78% mortality, whereas S. weiseri and S. carpocapsae killed 50% and 56% of the larvae, respectively. Both strains of H. bacteriophora species caused less than 50% mortality. Except for S. feltiae, the majority of infected medflies died as prepupae or pupae within the puparia. More than 90% larval mortality was recorded at 200 and 400 IJs/cm² for S. feltiae. None of the nematode isolates infected the medfly pupae within the puparia. In pot experiments containing soil, S. feltiae caused 96% and 97% mortality at 100 and 200 IJs/cm², respectively. In pot experiments with grass present, more than 94% mortality was obtained in the presence of grass roots.     

Nematicidal Activity of Chrysanthemum coronarium

Organic amendments and green manure are potential alternatives to the harmful chemical control means currently used against plant-parasitic nematodes. In this work, Chrysanthemum coronarium was applied to the soil as a green manure to control the root-knot nematodes Meloidogyne incognita and M. javanica. Chrysanthemum coronarium significantly reduced nematode infection of tomato roots and improved plant-top fresh weight, both in the greenhouse and in microplots. Other green manures, derived from Anthemis pseudocotula, wild chickpea (Cicer pinnatifidum), Geranium spp. and wheat, were not as effective as C. coronarium. Chrysanthemum coronarium, retained its nematicidal activity even when applied as a dried material. Only mature C. coronarium plants, in their flowering stage, exhibited nematode control activity, but the green plant parts were more effective than the flowers. An aqueous extract of C. coronarium exhibited in vitro, nematostatic activity towards M. incognita and M. javanica second-stage juveniles and inhibited their hatching from eggs and egg-masses; its nematostatic activity was expressed also against other phytonematode species such as Heterodera avenae and Pratylenchus mediterraneus, but did not affect the beneficial entomopathogenic nematode Steinernema feltiae.     

The impact of resistant and susceptible wheat cultivars on the multiplication of Heterodera filipjevi and H. avenae in parasite‐infested soil

Cereal cyst nematode (CCN) severely threatens wheat production in many regions of China. Cultivars susceptible to CCN are the main reason for its spread. This study was initiated to determine whether wheat cultivars conferring different resistance levels are the dominant determinants of CCN populations in the rhizospheric soil. Field experiments were conducted at two locations in Henan province, China, where high populations of Heterodera filipjevi or H. avenae have occurred, during the growing seasons of 2010/11 and 2011/12. Conventional enumeration of white female nematodes on the plant roots, reproductive factor (Rf) and a molecular diagnostic approach, PreDicta B test, a soil testing service based on a sensitive quantitative PCR technique, were used to determine the nematode populations in the rhizospheric soils of seven common wheat and durum wheat cultivars with different reactions to CCN. The resistant responses to CCN conferred by durum wheat Wascana and Wakooma and common wheat Madsen were effective against both H. filipjevi and H. avenae and resulted in significantly fewer nematodes (<5 females) on the roots, and lower Rf. Those cultivars were effective in limiting nematode propagation, resulting in fewer CCN eggs in their rhizospheric soils. Taikong 6 conferred moderate resistance (5–10 females) to both Heterodera species. Tianmin 668 only showed resistance to H. avenae. Aikang 58 and Wenmai 19 were susceptible to both CCN species, which facilitated increases in the nematode populations. These results demonstrate that the reactions to CCN of wheat genotypes have obvious impact on the propagation of nematodes.     

Suitability of weed species prevailing in Spanish vineyards as hosts for root-knot nematodes

Commercial vineyards in southern Spain were surveyed and sampled during October to December 2004 to determine the extent to which common weeds present were suitable hosts of root-knot nematodes infesting soils of those vineyards. Seven weed species commonly growing in grapevine soils in southern Spain were found infected by either Meloidogyne incognita or M. javanica: Amaranthus retroflexus (redroot pigweed), Anchusa azurea (ox-tongue), Chenopodium album (goosefoot), Erodium moschatum (musk stork’s bill), Malva rotundifolia (low mallow), Sinapis alba (white mustard), and Solanum nigrum (black nightshade). The host suitability of the weeds to root-knot nematodes was evaluated on the basis of root galling severity and nematode population densities in soil and roots. Also, the host–parasite relationship in these naturally Meloidogyne-infected weeds was examined. All the weed species in the study were considered suitable hosts for M. incognita and M. javanica because: (a) high Meloidogyne spp. populations occurred in roots and surrounding soil of the weed species; (b) the severity of root galling was high, and (c) well-established permanent feeding sites were observed in the histopathological studies of infected root tissues. In addition, this study presents the first reports of S. alba and A. azurea as hosts for M. incognita, and of E. moschatum as a new host for M. javanica, thus increasing the list of reported weed hosts for Meloidogyne spp. These results indicate that noticeable population densities of M. incognita and M. javanica can be maintained or increased in these weeds, at population levels higher than those previously reported for the same nematodes infecting grapevine roots. The weeds infesting vineyards thus represent an important source of inoculum of Meloidogyne spp., and furthermore may act as reservoirs of these nematodes which can be disseminated within or among vineyards by agricultural operations.     

Effects of antagonistic fungi,plant growth-promoting rhizobacteria,and arbuscular mycorrhizal fungi alone and in combination on the reproduction of <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> and growth of tomato

Antagonistic fungi (Aspergillus niger CA and Penicillium chrysogenum CA1), plant growth-promoting rhizobacteria (PGPR) (Burkholderia cepacia 4684 and Bacillus subtilis 7612) and AM fungi (Glomus intraradices KA and Gigaspora margarita AA) were assessed alone and in combination for their effects on the growth of tomato and on the reproduction of Meloidogyne incognita in glasshouse experiments. Application of antagonistic fungus, PGPR, or AM fungus alone or in combination significantly increased the length and shoot dry mass of plants both with and without nematodes. The increase in shoot dry mass caused by Gl. intraradices KA in plants without nematodes was greater than that caused by PGPR or antagonistic fungi. Similarly, the increase in shoot dry mass caused by Bu. cepacia 4684 in plants with nematodes was greater than that caused by P. chrysogenum CA1. Application of Bu. cepacia 4684 caused a 36.1% increase in shoot dry mass of nematode-inoculated plants followed by Ba. subtilis 7612 (32.4%), A. niger CA (31.7%), Gl. intraradices KA (30.9%), Gi. margarita AA (29.9%) and P. chrysogenum CA1 (28.8%). Use of Bu. cepacia 4684 with A. niger CA caused a highest (65.7%) increase in shoot dry mass in nematode-inoculated plants followed by Ba. subtilis 7612 plus A. niger CA (60.9%). Burkholderia cepacia 4684 greatly reduced (39%) galling and nematode multiplication, and the reduction was even greater (73%) when applied with A. niger CA. Antagonistic fungi had no significant effect on root colonization caused by AM fungi. Applying Bu. cepacia 4684 with A. niger CA may be useful in the biocontrol of M. incognita on tomato.     

Compounds inhibitory to nematophagous fungi produced by Bacillus sp. strain H6 isolated from fungistatic soil

Soil fungistasis can cause inconsistent control by nematophagous fungi of plant-pathogenic nematodes in field situations. Recent studies have shown that production of fungistatic compounds by bacteria was the principal explanation for soil fungistasis. The culture filtrate of Bacillus sp. strain H6, a strain representative of the dominant colony types isolated from fungistatic soils, showed strong inhibitory activity against nematophagous fungi by inducing unusual swelling in the conidia and the germ tubes of nematophagous fungi, thereby preventing the fungi from proliferating. This inhibitory mechanism is novel in comparison with other known mechanisms. Antifungal activity of the culture filtrate of strain H6 was maximal after culture in Luria Bertani (LB) broth (pH 7.0) at 36°C for 36 h. The inhibitory effect of the compounds produced by Bacillus sp. strain H6 was not significantly influenced by pH, and the inhibitory compounds in the culture filtrate were thermostable. After being partially purified by thin layer chromatography (TLC) on silica gel plates, characterization by colour reactions and positive fast atom bombardment mass spectrometry indicated that the inhibitory compounds showed similarity to iturin A group compounds.