Plant parasitic nematodes

A survey to study the presence and distribution of plant-parasitic nematodes associated with grape (Vitis vinifera L) was conducted in Ismailia vineyards. A totals of 460 plant and soil samples were collected from vineyards of five different localities in Ismailia governorate viz. " Ismailia, El-Tall al Kabir, Abou-Swair, El-Kantara Sharq, and El-Kassasen". The results obtained from the survey showed that the population varied in their frequency and distributed among surveyed vineyards. There were eleven plant parasitic nematode genera found associated with common grape plants in Ismailia Governorate. These genera were Meloidogyne spp.,

Endophytic bacteria isolated from internal tissue of surface sterilized jatropha curcas L. leaves, stem, frouts and roots. Collected from field plots produced secondary metabolites in nutrition broth media (NB) that were highly toxic to Meloidogyne spp. especially strains SJS54 followed by RJS175 were highly active among 23 strains producing secondary metabolite toxic to Root-knot nematodes. The mechanism of action of the toxic metabolites produced by the non-pathogenic strains SJS54 and RJS175 with proven biological control of Meloidogyne spp in Syracuse glass experiments was investigated. These metabolites reduced Meloidogyne spp mobility within 6-12 h of exposure. After 6 h, approximately ±50% of juveniles was inactivated. But with exposure of 12 h 80-90% of the juveniles were dead. The SJS54 and RJS175 toxicity were highly effective towards migratory parasites. Most of nematodes have been shown body wall degradation caused by hydrolytic enzymes produced by endophytic bacteria, a...

The Root-knot nematode Meloidogyne incognita is one of the most economically important pests causing severe damages and losses in a wide variety of crops. Its management using resistant plant cultivars or appropriate crop rotations is an effective and environmentally friendly method. However, this strategy requires an accurate and rapid discrimination of the populations regarding virulence on resistant cultivars or crop-/cultivar dependent aggressiveness. Here we report development of a polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) as a molecular tool to detect differences in DNA sequences or mutations of various genes to compare the distribution of variants of the effector gene msp1 between seven populations/races originated from different countries. Sequencing of cloned PCR products revealed five msp1 variants from these populations which were distinguishable in their reproduction on five host plants. PCR-DGGE facilitated the discrimination among these populations and/or races based on these msp1 variants that were detected by five clear bands at different positions. DGGE for replicated pools of juveniles from the seven populations revealed ten variants of msp1. A correlation between the presence of a particular gene variant and the reproductive potential on particular hosts was not evident. Especially race 3 showed substantial variation within the population. DGGE fingerprints of msp1 tended to cluster the populations according to their reproduction rate on pepper. The developed method could be useful for analyzing population heterogeneity and epidemiology of M. incognita. Root-knot nematodes, Meloidogyne spp., generally are the economically most important nematode pests that parasitise agri-and horticultural crops in South Africa. The aims of the study were to i) identify Meloidogyne spp. individuals that infect roots/tubers/pods of crop plants received for diagnostic analyses and from research sites across South Africa using molecular and morphological identification techniques and ii) determine the reproductive potential of Meloidogyne species populations identified during the study in a greenhouse experiment (randomised complete block design with six repli-cates). Deoxyribonucleic acid (DNA) was extracted from mature females obtained from infected, below ground parts of crops and subjected to polymerase chain reaction (PCR) analyses. For the reproduction-potential study, 1000 eggs and second-stage juveniles (J2) of the respective Meloidogyne spp. populations identified were inoculated on roots of a susceptible tomato cultivar Floradade. Nematode parameters assessed 56 days later included egg-laying female indices, egg and J2 numbers and reproduction factors/root system. Meloidogyne incognita and M. javanica proved to be the predominant species that infected maize, potato and soybean crops, while the emerging M. enterolobii (= M. mayaguensis) have also been identified from pepper and guava roots. Other unknown species have also been detected and are currently being identified. The reproduction potential of the various Meloidogyne spp. populations differed substantially within and among species. Positive identification of M. enterolobii, which is easily confused with M. incognita in terms of its morphological identification will contribute towards research aimed at determining the distribution, life cycle and path-ogenicity of this pest. This study is ongoing and knowledge generated will benefit the research fraternity as well as producers and ultimately consumers. A gram negative strain bacteria was isolated from healthy pepper plants grown on naturally infected soil by the plant pathogenic root knot nematode Meloidogyne located at the Agricultural Experimental Farm Station, Faculty of Agriculture, Ismailia. In the dual culture bioassay, an experiment was performed in Petri dishes to examine the effect of the isolated bacteria against egg-masses hatching of the root knot nematode Meloidogyne javanica. Results revealed that there was a 218

An investigation was carried out for two consecutive years, 2016 and 2017, to test the efficacy of different pre sowing nursery treatments with two fumigants viz., dazomet (@ 40g/ m) and metham sodium (@ 14ml/ m), one organic formulation i.e., jeevamrit (@ 10%), and two bioagents viz., Trichoderma viride (@ 50g/ m) and Pseudomonas flourescens (@ 50g/ m), along with untreated control, against root knot nematodes (Meloidogyne incognita) infesting tomato nursery. The results revealed that maximum percent germination and shoot lengths were of jeevamrit, dazomet and metham sodium followed by T. viride and P. flourescens treated seedlings, all being better than control. In reference to nematode soil populations and root gall index, dazomet and metham sodium had better efficacy in comparison to T. viride, P. flourescens, jeevamrit and control. There was no significant difference in nematode soil population between jeevamrit and control.

El objetivo del presente estudio fue realizar una prospección de nematodos fitoparásitos asociados al cultivo de cebolla en el valle de Asunción Mita y relacionarlos con la presencia de Fusarium oxysporum. Se realizaron muestreos de suelo en diferentes puntos del valle de Asunción Mita en parcelas con cebolla. En cada punto se tomaron 10 submuestras para obtener una muestra compuesta de aproximadamente 1-2 Kg. Se trasladaron las muestras al laboratorio de patología vegetal en el Campus Central de la Universidad Rafael Landivar y se utilizó el método de tamizado y centrifugado con flotación en azúcar para la extracción de los nematodos. La identificación de géneros de los especímenes extraídos se realizó con base a la clave de identificación para hembras de Mai y Mullin (1996). Se cuantificó la incidencia de F. oxysporum en parcelas matrices de un metro cuadrado colocando un marco de madera en tablones con plantas sintomáticas y se contó la cantidad de plantas afectadas entre en total de plantas que cabían dentro del marco. Los resultados indican la presencia de diez géneros de nematodos: Aphelenchoides, Helicotylenchus, Meloidogyne, Paratrichodorus, Pratylenchus, Scutellonema, Hoplolaimus, Criconemoides, Tylenchorynchus y Tylenchus. En promedio se encontraron con mayor presencia el género Helicotylenchus (26.5 nematodos/100cc de suelo) y menor cantidad los géneros Meloidogyne y Paratrichodorus (1 nematodo/100cc de suelo). Se pudo observar una incidencia de F. oxysporum distribuido en parcelas del valle que va de 10%-50%. Los resultados indicaron que no existe  relación de la incidencia de F. oxysporum con la cantidad total de nematodos, lo cual se concluye que los nematodos fitoparásitos en el valle de Asunción Mita, no son factores inductivos de esta enfermedad.

Global agriculture is currently challenged to provide increasing supplies of food for a growing population due to the changing trends of insect attacks and climatic conditions. There is overwhelming evidence that the use of chemical pesticides have created many other serious problems like ecological backlashes in pest species, environmental pollution and degradation, threat to biodiversity conservation, loss of beneficial fauna (predators, parasites, pollinators etc) and human beings in particular. Keeping in view, the new generation of pesticides have attracted attention in pest management in recent years, Biorational pesticides as “third-generation pesticides” are derived from some natural source and impose minimum or no adversarial threats on the environment or beneficial organisms. Some examples of biorational pesticides are the microbial pesticide Bacillus thuringiensis (Kurstaki), neonicotinoids, avermectins, phenlpyrazoles, spinosyns, pyrroles, oxadiazines and various groups of insect growth regulators. With a narrower target range of pests, they also tend to have a more specific mode of action. This new class of pesticides are often designed to control a pest population to a manageable level rather than completely eradicate a target pest. These products have great potential for replacing the persistent conventional insecticides, confirming effective cost-benefit ratio, tackling ecological backlashes and ensuring food security with safe environment. The field of biorational pesticides is deep; consequently they are a source of both optimism and concern. However, these bio-products are not only winning the reliability of the market and end user; but also demonstrating their worth and potential in sustainable integrated pest management (IPM) program.

Національний університет біоресурсів і природокористування України НДІ фітомедицини, біотехнологій та екології Факультет захисту рослин, біотехнологій та екології Агенство з трансферу технологій, освіти і науки daRostim Німецький приватний інститут прикладної біотехнології daRostim БІОЛОГІЧНО АКТИВНІ ПРЕПАРАТИ В РОСЛИННИЦТВІ НАУКОВЕ ОБҐРУНТУВАННЯ-РЕКОМЕНДАЦІЇ-ПРАКТИЧНІ РЕЗУЛЬТАТИ Матеріали XV Міжнародної науково-практичної конференції Київ, 25-29 червня 2019 року

Nematoda puru akar (NPA) Meloidogyne spp. merupakan salah satu patogen penting pada berbagai tanaman hortikultura dan beberapa tanaman pangan di Indonesia. Infeksi NPA mampu menurunkan kuantitas dan kualitas panen, bahkan menyebabkan gagal panen apabila terjadi sinergisme dengan patogen lain. Penggunaan pestisida kimia sintetis untuk mengendalikan Meloidogyne spp. dan patogen lain di kalangan petani sering melebihi batas pemakaian normal. Selain menyebabkan kerusakan dan pencemaran lingkungan, penggunaan pestisida kimia sintetis dapat menyebabkan gangguan kesehatan pada petani, meninggalkan residu kimia yang berbahaya bagi konsumen, dan menurunkan jumlah mikroba dan mikro fauna yang berperan penting dalam pertanian berkelanjutan. Masalah tersebut dapat ditekan tingkat kejadiannya dengan mengurangi penggunaan pestisida kimia sintetis. Terdapat banyak pilihan cara untuk mengendalikan NPA tanpa menggunakan pestisida kimia sintetis, yaitu dengan pola tanam polikultur, memanfaatkan tanaman antagonis, teknik biofumigan, penggunaan ekstrak nabati (pestisida nabati), penggenangan sawah, aplikasi agens antagonis, dan aplikasi metabolit sekunder dari mikroba. Cara-cara tersebut diketahui efektif menekan populasi NPA pada berbagai skala percobaan, dan tidak menimbulkan masalah lingkungan. Beberapa teknik aplikasi seperti pola tanam polikultur, dan aplikasi agens antagonis diketahui tidak hanya menekan populasi NPA dan mengurangi kerusakan akibat infeksi NPA, namun juga mampu meningkatkan hasil panen. Teknik-teknik pengendalian patogen yang ramah lingkungan perlu dilakukan untuk mewujudkan swasembada pangan melalui pertanian berkelanjutan.

Kata kunci: Meloidogyne spp., nematoda puru akar, pengendalian

Plant parasitic nematodes (PPNs) are important constraints in
crop production. On global basis plant parasitic nematodes are known
to cause estimated average crop losses of about12.3 per cent (Sasser

and Freckman, 1987). In developing countries having tropical and sub-
tropical climates, the losses due to plant parasitic nematodes have been

estimated at 14.6% as compared to 8.8 % in developed nations. In
another estimate plant parasitic nematodes are known to cause losses
equivalent to US$173 billion per year globally in major crops (Elling,
2013). Recent crop assessment carried under All India Coordinated
Research Project (AICRP) on nematodes in Agriculture revealed 21.3
per cent average crop losses in India amounting to RS 102,039.79
million (1.48 billion USD) annually (Kumar et al, 2020). The losses in
19 horticultural crops have been assessed at Rs. 40,224.98 million.
Morphologically, nematodes are triploblastic, bilaterally symmetrical
and cylindrical invertebrates without cilia and true coelom. Plant
parasitic nematodes bear a needle like structure in their mouth known
as stylet/ spear which is used to puncture plant roots and suck the cell
sap. PPNs feed on plant roots or other parts ectoparasitically or
endoparasitically depending upon their mode of parasitism. Further,
the endoparasitic nematodes are more damaging than ectoparasites.
PPNs are considered as hidden enemies as majority of them are soil
inhabiting; the above ground symptoms produced are mainly
nonspecific which are generally go unnoticed and confused with
nutritional deficiencies. Besides causing direct damage to plants, PPNs
also form interactions with other plant pathogens resulting in disease
complexes (Atkinson, 1892). Plant damage is increased manifold in
these disease complexes. PPNs are also known to transmit soil borne
plant viruses. The intensification of agriculture to meet the demands of
growing human population has resulted in increase in nematode
problems.

Nematology Nematology is the scientific discipline that deals with the study of nematodes. Nematology is conventionally concerned with the study of phyto-nematodes which parasitize plants and are of economic importance to agriculture, horticulture and forestry. "Nematode" is a greek word (nema = thread, oides = form) i.e. thread like organisms as they look like tiny threads moving under microscope. Nematodes Nematodes may be defined as bilaterally symmetrical, triploblastic, pseudocoelomic, unsegmented invertebrates with four hypodermal chords, a tri-radiate oesophagus, circum oesophageal nerve ring and a definite tail but lack circulatory muscles and specialized organs for locomotion and respiration. Nematodes belong to phylum Nematoda (also called Nemathelminthes). Synonyms: Threadworms, eelworms (serpentile eel like body), round worms, nemas, paudh/padap, krimi etc.

Three field trials was conducted in Ismailia governorate, Egypt during the seasons 2009, 2010 and 2011 to evaluate the efficacy of twelve granular and liquid chemical nematicides produced by different companies and three bio-nematicides on populations of root-knot nematodes, Meloidogyne javanica infesting banana and grape and citrus nematodes, Tylenchulus semipenetrans infesting orange as well as crop productivity. Among all chemical nematicides applied on banana, cure (oxamyl SL 24%) treatment at the rate of 15 ml/mat twice surpassed other treatments in reducing rate of nematode build-up with value of 0.24, followed by nemacur G 10% at 30 g/mat and vydate SL 24 % at 15 ml/mat twice treatments with the same value of 0.31 each, whereas nemathorin G 10 % treatment at 3 g/m 2 recorded the lowest reduction percentage of the same criterion with value of 0.50. Application of nemacur G 10% and vydate SL 24% gave the highest increment percentage in crop productivity with value of 65.6 % each, followed by cure treatment (59.2 %). Nemathorin treatment at 12.5 kg/feddan accomplished the best results in improving the production of orange fruits and reducing rate of T. semipenetrans build-up as well where the percentage increase value of productivity averaged to 91.8% and the reduction percentage value of nematode build-up averaged to 82.9%, but no significant differences were noticed among all nematicides tested. The highest reduction in rate of M. javanica build-up was obtained when grape trees treated with vydate 24% SL at the rate of 5 L/feddan twice that amounted to 0.29 as well as the highest percentage increase of crop production with value of 90.2%. It was clear that chemical nematicides gave more efficient control of nematode population density than the bio-nematicides for an immediate control on grape trees. Among all tested bio-nematicides, QL Agri 35 decreased rate of M. javanica build-up and increased grape production to the maximum with values averaged to (0.57 and 7.50).

A survey was conducted in Udaipur, Chittorgarh, Bhilwara, Kota and Bundi districts of Rajasthan to know the occurrence of entomopathogenic nematodes during 2007-08. Total 105 soil samples were collected from different localities by random sampling technique. The soil samples were baited twice using last instar larvae of the greater wax moth. The occurrence of entomopathogenic nematodes in the soil samples was established on the basis of mortality of G. mellonella larvae. Out of 105 soil samples, three samples were positive for EPNs in these surveyed districts of Rajasthan.

A system, method, and apparatus for quantification of pre-determined particles in a soil or feces sample with certain automated steps. In one aspect it includes an input station for inputting a soil or feces sample; a sieving/filtering station for separating soil or feces from particles or particle carriers, and/or separating particle carriers from particles; and a collection station for receiving the extracted particles. It can include quantification of the collected sample with an imaging station to digitally image the particles and recognize and count the particles collected. A mechanism can mechanically move the filtered particles from the sieve/filter station to the collection station. A controller can be programmed to automatically control at least certain functions of the mechanism. An optional feature includes acquisition of chemical, biological, physical, or other parameters of the sample with one or more sensors positioned or positionable at the sample.

Plant-parasitic nematodes are economically important pests of banana (Musa spp.) and compromise its productivity. Controlling nematode populations and good agronomic practices are prerequisites for a good yield. A survey of farmers in the Rusitu Valley, Zimbabwe, was conducted to assess their current banana production practices and knowledge of plant-parasitic nematodes associated with banana. Respondents were selected using systematic sampling from three wards. Most farmers (61.9%) grew bananas as a monoculture, and 38.1% intercrop banana with other crops. All the farmers neither rotated banana with other crops nor practiced pest and disease control measures, and only 11.4% apply a fertiliser to their banana crops. Most (82.9%) farmers in Rusitu Valley had little or no knowledge of plant-parasitic nematodes that damage bananas. Sound extension programme in Rusitu Valley should educate farmers on the importance of managing plant-parasitic nematodes and using better banana crop production practices.

The bacteria Xenorhabdus nematophilus and Xenorhabdus bovienii, isolated from the entomopathogenic nematodes Steinernema carpocapsae and Stein-ernema feltiae respectively, were assayed against three stages of the insect Phaedon cochleariae (adults, larvae and pupae). The aim of this work was to assess the use of bacteria isolated from entomopathogenic nematodes (EPN) against insect in an attempt to propose new approaches of biocontrol insects. Experiments had the broad objective to investigate the mortality of P. cochleariae when exposed to the bacteria suspensions both in vivo and in vitro. High mortality was obtained in all experiments when the bacterial suspensions were applied to the insects. There was no significant difference between the mortality of the three different developmental stages of P. cochleariae, which indicates that the bacteria were high virulent to P. cochleariae. The mortality of P. cochleariae in vivo experiments was found to be lower than in vitro and the mortality in the assessment with X. bovienii in vivo was found higher than in X. nematophilus. In addition, a small experiment was carried out using Galleria mellonella larvae. The results from this experiment revealed the larvae of the larger G. mellonella took longer to die than the larva of P. cochleariae suggesting that body size is important. Also, it seems that bacteria isolated from entomopathogenic nematodes could be used successfully to control foliar insect pests.

A pot experiment was conducted in order to study the impact of time application of four animal manures i.e. chicken, horse, farmyard and pigeon manures and town refuse added separately either pre or post planting treatments comparing to oxamyl on tomato plant cv. Castle Rock infected with M. incognita under greenhouse conditions (25±3 o C). Results revealed that all tested materials clearly enhanced plant growth characters to great degree and reduce gall and eggmass numbers. Plant receiving pigeon manure as pre-planting or post planting applications at the level of 5g/plant obviously surpassed other tested animal manures and town refuse in the increment values of plant length (85.7 and 71.4%), total plant fresh weight (66.4 and 37.9%) number of flowers (100 and 100%), fruits (300 and 400%), and branches/plant(100 and 300%) and shoot dry weight (42.9 and 38.1%), as well as achieved the highest reduction percentage for number of root galls(89.1 and 81.5 %), eggmasses (90 and 84.2%), for pre or post planting applications, respectively followed by farmyard manure where as chicken treatment as a pre-planting soil amendments exhibited the lesser values in this respect. All treatments obviously gave sizeable percentage increase values of nitrogen (N), phosphorus(P), potassium(K), organic matter(O.M), and organic carbon(O.C) in leaves of tomato plant. The pigeon or chicken manures showed also very narrow C/N ratio (12.6:1) or (14.3:1), respectively, in this work. Among the tested materials, pigeon manure ranked first in percentage increase values of chemical components of leaves as pre-planting application, whereas chicken manure showed this position as post-planting treatment.

Weeds with allelopathic properties can influence vegetational patterns in agroecosystems. Wild mustard (Brassica kaber), has the potential to form and release biologically-toxic isothiocyanates into the soil. The effect of mustard management on the incidence of soil-borne pathogens on corn was investigated. Under field conditions a significant negative correlation was found between plant-parasitic nematode numbers and mustard biomass. The most abundant parasitic nematode genera were: Meloidogyne, Tylenchorhynchus, Helicotylenchus, and Pratylenchus. The incidence of root- pathogenic fungi (Fusarium spp., Rhizoctonia solani, and Pythium aphanidermatum) in corn was low. No differences in soil fertility or moisture were found. Greenhouse experiments confirmed these results and showed no differences in corn-root biomass that could explain the differences in nematode populations. Secondary compounds exuded by mustard roots were hypothesized to explain the phenomenon. Neither Water and acetone extracts of mustard roots or carbofuran alone affected nematodes movement. However, after exposed to root extracts carbofuran had a highly toxic effect on nematodes of the same genera as mentioned above. Allyl, benzyl and phenylethyl isothiocyanates, either as volatiles or in solution, were also toxic to the same nematodes genera. Isothiocyanates volatiles were more toxic to nematodes than they were in sterile soil. Soil microorganisms seem to enhance the toxic effect on nematodes. Volatiles of isothiocyanates, singly and mixed, inhibited hyphal growth of three isolates of Pythium aphanidermatum in a dose-dependent response. Pythium isolates were obtained, one each, from corn and mustard roots growing together in greenhouse, and a third one from an area near Mexico City where the wild mustard is not common. Isothiocyanate mixtures had additive toxic effect on isolates from corn and mustard, and a synergistic toxic effect on the Mexican isolate. When fully inhibited by allyl isothiocyanate, a bacterial colony, identified as Pseudomonas, grew from all Pythium inocula. The nature of the infection of P. aphanidermatum by Pseudomonas was assessed. All results are discussed in the context of controlling soil-borne pathogens through the management of weeds with the ability to release allelochemicals into the soil.

Plant-parasitic nematodes of the genus Ditylenchus infect a wide range of plants causing serious economic losses in the temperate climatic zones. Among them, two species, Ditylenchus destructor and D. dipsaci, able to infect potato, are the most destructive and are included in the lists of quarantine and regulated pests in many countries. A proper diagnostics of these species is very relevant for timely and reliable evaluation of the quality of seed potato and the phytosanitary state of potato fields. Based on ITS1-5.8S-ITS2-28S rDNA sequences, test systems for detection of these two species and also for D. weischeri, a confounding species, were developed suitable for the use in complex qPCR micromatrices (microarrays) under a standardised real-time PCR protocol. The test systems demonstrated high specificity and reproducibility and satisfactory analytical sensitivity (5 juveniles for D. destructor and D. dipsaci and 50 juveniles for D. weischeri) suitable for practical use. The detection limit was 8-10 pg of DNA for D. destructor and D. dipsaci and 77 pg of DNA for D. weischeri. Working efficiency of the developed test systems after their freeze-drying on preserved silicon micromatrices was not significantly reduced. Due to the simplicity of the use of qPCR micromatrices, flexibility of their structure, and reduced time of analysis, the developed test systems may be recommended for the use in field laboratories including possible cases of combined infection, when the presence of harmful Ditylenchus species is masked by symptoms caused by other pathogens and may be overlooked.

The aim of the study was to evaluate the biocontrol potential of Trichoderma harzianum against Meloidogyne incognita and decipher mechanisms of induced systemic resistance and disease suppression in tomato grown in net house conditions. The fungal biocontrol agent T. harzianum UBSTH-501 was evaluated against M. incognita on dual plate under in vitro conditions and in planta under nethouse conditions. The results of in vitro parasitism on dual plates showed that T. harzianum causing infection on the eggs and juveniles of root-knot nematode, whereas, in planta assay showed that plants treated with talc based bioformulation T. harzianum UBSTH-501 exhibited manifold increase in the accumulation of total chlorophyll and enzymes, viz. chitinase, phenylalanine ammonia lyase (PAL) and peroxidase which is known to confer systemic resistance in tomato against M. incognita resulting into decreased nematode population and disease severity. Results revealed that T. harzianum UBSTH-501modulated phenylpropanoid pathways led to enhanced accumulation of defence related mediator molecules and enzymes in tomato resulted in disease suppression to a significant extents.

The burrowing nematode of banana, Radopholus similis, is difficult, if not
impossible, to control effectively. The lack of resistance in commercially acceptable cultivars and the perennial nature of the crop limit management options. Multiple- cycle treatment with systemic nematocides is presently the only effective tool available for reducing damage and sustaining yield. Most of these compounds are not toxic to the burrowing nematode, but only inhibit nematode activity for short periods of time. The repetitive application of non-fumigant nematocides has led to an increase in the rapidity of their biodegradation. From environmental, toxicological and consumer viewpoints, this type of pest management is unacceptable. Many attempts have been made to improve the situation with little or no impact to date. Many nematocides have been or are being removed from the market, and new replacement compounds have not been developed. Resistance has never been detected in commercial banana cultivars, and genetically modified cultivars are still unacceptable to a large number of banana consumers. Microbial- based strategies are considered to be a promising alternative approach to nematode management in banana. Progress can only be made if research and crop production systems are streamlined to support new management concepts. The following are discussed: 1) biological enhancement of planting material with fungal endophytes; 2) treatments with multiple nematode antagonists with diverse modes of action; 3) importance of induced systemic resistance in biological control systems; and 4) use of molecular tools to detect effective antagonists.

This study was conducted to find the incidence of root knot nematode (M. javanica) in winter weeds of tomato at the root knot infested areas of Dargai and Jabban in Malakand division of Khyber Pakhtunkhwa province of Pakistan. For this purpose survey was conducted on 20 fields of each region and the fifteen weed plants were checked for galls caused by M. javanica on their roots. Weeds samples from these regions were bagged and labeled properly and brought to the Department of Weed Science, The University of Agriculture, Peshawar Pakistan for identification. The nematode was also identified by perennial pattern. Galls were observed on commonly occurring fifteen weeds. Incidence of M. javanica on Cannabis sativa was at the range of 9.9-11.9%, while it was 33.3% on Digitaria sanguinalis. The highest incidence of M. javanica were recorded on the roots of Fumeria indica (21.05-43.47%). Moreover the incidence of M. javanica on Rumex crispus and Melilotus indica was 22.72% and 22.13% respectively. However no galls (No incidence) of M. javanica were found on Asphodelus tenuifolius. Our finding demonstrates that M. javanica is a damaging pathogen of winter weeds of tomato in Malakand division of Khyber Pakhtunkhwa province of Pakistan.

The suppressive effects of six species of entomopathogenic nematodes (EPNs) against Meloidogyne incognita infected tomato were assessed in two separate trials under greenhouse conditions. The tested EPN species were Heterorhabditis bacteriophora, H. megidis, H. zealandica, Steinernema feltiae, S. glaseri and S. riobravae. In the first trial , EPNs were applied as a liquid suspension of alive or dead infective juveniles (IJs) at rate of 5000 IJs/plant in sandy and clay soils, while in the second trial, two Galleria mellonella infected cadavers of six-day old per plant were used in sandy soil. In the two experiments, application of EPNs was accomplished simultaneously with inoculation of M. incognita. Carbofuran as a standard synthetic nematicide was used as comparison treatment at recommended rate of 0.2 g/plant. Control treatments received only water and M. incognita at rate of1000 IJs/ plant. Two months after inoculation, galling (as indicated by number of galls/plant) and reproduction (as indicated by number of egg masses /plant) as well as damage (as indicated by fresh and dry weight of areal parts) were assessed. Data showed that, treatment of carbofuran surpassed all other treatments in minifying galling and reproduction of M. incognita in sandy and clay soils. On the other hand, curative applications of alive or heat-killed IJs significantly (P≤ 0.05) diminished gall formation and egg mass production in tomato roots with slightly amelioration in fresh and dry weight of tomato shoot. Steinernematid species were more comparatively effective than heterorhabditid ones. General means for number of galls and egg masses for steinernematid species were 52.50 and 25.33 with percent reduction of 55.45 and 64.49 %, respectively. Whereas, the parallel values for heterorhabditid species were 69.67 and 36.33 with percent reduction of 40.88 and 49.07%, respectively compared to treatment of M. incognita alone. Treatments of alive IJs overwhelmed those of dead IJs in decreasing number of galls (with percent reduction of 56.53 and 39.79% respectively) and egg masses (with percent reduction of 63.16 and 50.39 %, successively). Moreover, utilization of two G. mellonella infected cadavers markedly lowered number of galls and egg masses and insignificantly (P≤ 0.05) improved plant growth parameters to certain extent. General means of percentage reduction in galls and egg masses were 58.46 and 54.74%, consecutively.

ABSTARCT A pot trail was conducted in order to study the influence of four organic acids i.e. salicylic , citric , oxalyic and humic acids at two concs.0.1& 0.05% in comparison with oxamyl against Meloidogyne incognita infecting tomato cv.9065FI under greenhouse(24±2C •).Results indicated that all tested treatments clearly improved tomato plant characters and reduced nematode development. Among the organic acid tested, salicylic acid at the two concs. surpassed other application in values of percentage increase of plant length (23.37&40.54%), number of leaves (28.03&55.3%), number of branches (16.7&54.2), total plant fresh weight (32.3&57.23%) and shoot dry weight (25.2&33.84%), and achieved the highest percentage reduction values of final nematode population(78.08%), number of egg-masses (89.47%), respectively. In the mean time, oxalic acid applications gave the least values in this respect comparing to the check. Salicylic acid and humic acid at 0.05% since their eggmasses indices were in par (2) vs (4) for nematode alone. Among tested concentrations , 0.1 g/L of such organic acids was higher in values of N, P , K , C , O.M and total chlorophyll as compared to nematode alone , whereas the concentration of 0.05g/L of the four organic acids ranked first in percentage increase values of total phenol which were amounted to 8.39 , 6.36 , 4.43 and 2.68% for salicylic , citric , oxalic and humic acids comparing to nematode alone , respectively.

Four untraditional materials were tested to control Meloidogyne incognita juveniles under laboratory conditions, i.e. magnetic iron, tourmaline, phosphate granules and saponines extract compared to traditional nematicide fenamifos 40% EC. These materials were tested at the concentrations of 500, 1000 and 2000 ppm directly against M. incognita juveniles in Petri dishes and the results were obtained after 24, 72 and 144h. All tested materials showed nematicidal effect on nematode juveniles and their mortality increased with increasing concentration and exposure period. fenamifos 40% EC was the most effect one on juvenile activity followed by saponines extract, phosphate granules, tourmaline, and finally magnetic iron. Numbers of active juveniles were 50.0, 90.34, 260.67, 280.34 and 460.67 juveniles/1 ml, respectively, compared to control 850.11 juveniles /1ml at a concentration of 2000 ppm after 144 hours of treatment. On the other hand, fenamifos 40% EC was the most effective one in inhibiting egg hatching followed by saponines extract, phosphate granules, tourmaline and magnetic iron in the concentrations 500,1000 and 2000 ppm after 144 hours of exposure. Generally the tested eco-friendly materials showed nematicidal effect on juveniles and eggs of M. incognita under laboratory conditions and these materials should be tested under field or greenhouse conditions to show the effect on plant growth and nematode population.

Distilled water and acetone extracts from roots of Brassica kaber were tested in vitro as nematicides on J2 and J3 stage of nematodes recovered from soils from a field where barley and fava bean were growing. No effect on nematode motility was noted for up to 48 h. When carbofuran was added nematodes were rapidly (12 h or less) immobilized. Allyl, benzyl and phenylethyl isothiocyanates dissolved in acetone also showed strong nematicidal activity between 30-120 min, depending on concentration. Natural and sterile soil containing a known number of nematodes were used in bioassays to test the nematicidal properties of the extracts and volatiles from pure isothiocyanates. After exposure, a high number of nematodes became inactive in both natural and sterile soil, but numbers were greater in natural soil. The effect of root extracts was enhanced by carbofuran extract.

Five cowpea varieties viz Asomdwe, Asetenapa, Hewale, Nhyira and Videza, were evaluated for their reaction to plant parasitic nematodes under natural infestation. Meloidogyne incognita, Pratylenchus brachyurus, and Rotylenchulus reniformis were isolated from the rhizosphere of the cowpea varieties although each of the varieties reacted differently to these nematode species. The highest number of M. incognita was recovered from the rhizosphere of Asomdwe which was 83% higher than Asetenapa which recorded the least number of M. incognita. Similarly, there was a significant difference (P<0.05) between R. reniformis juveniles recovered from the rhizosphere of Hewale and all the other varieties. There was however no significant differences in the number of P. brachyurus recovered although Nhyira recorded the highest. Two of the nematode genera-Meloidogyne incognita and Pratylenchus brachyurus were extracted from the roots of the various varieties with varied population densities. The highest grain yield was recorded in Hewale. It out yielded Nhyira, Videza, Asomdwe and Asetenapa by (58, 49.8, 41.5 and 11.9)% respectively.

Virulence of root-knot nematodes, Meloidogyne spp., on tomato
bearing the Mi gene for resistance
MUWAFFAQ KARAJEH1
, WALID ABU-GHARBIEH2
and SAMEER MASOUD1
1Department of Plant Protection and Integrated Pest Management, Faculty of Agriculture,
Mu’tah University, PO Box 7, Karak, Jordan 2
Plant Protection Department, Faculty of Agriculture, University of Jordan, Amman, Jordan
Summary. Three species of root-knot nematodes, Meloidogyne javanica, M. incognita races 1 and 2, and M. arenaria race 2 occur in Jordan. These species and their races were identified using morphological characters, the North
Carolina differential host test, and SCAR-PCR. The virulence of 83 isolates belonging to Meloidogyne species and
races was assayed. The virulence assay was based on the isolate reproduction rate on a resistant tomato cultivar
Betterboy bearing the Mi gene for resistance, and was compared with that on the susceptible tomato cultivar Rutgers.
Three M. javanica isolates were highly virulent on the resistant cv. Betterboy as indicated by their high root gall
index (4.73) and high reproduction factor (3.73). The horticultural parameters (shoot and root fresh weights and root dry weight) were negatively correlated with the reproduction factor.
Key words: race, reproduction, susceptibility, SCAR-PCR.

Soybeans are an important crop for global food security. Every year, soybean yields are reduced by numerous soybean diseases, particularly the soybean cyst nematode (SCN). It is difficult to visually identify the presence of SCN in the field, let alone its population densities or numbers, as there are no obvious aboveground disease symptoms. The only definitive way to assess SCN population densities is to directly extract the SCN cysts from soil and then extract the eggs from cysts and count them. Extraction is typically conducted in commercial soil analysis laboratories and university plant diagnostic clinics and involves repeated steps of sieving, washing, collecting, grinding, and cleaning. Here we present a robotic instrument to reproduce and automate the functions of the conventional methods to extract nematode cysts from soil and subsequently extract eggs from the recovered nematode cysts. We incorporated mechanisms to actuate the stage system, manipulate positions of individual sieves using the gripper, recover cysts and cyst-sized objects from soil suspended in water, and grind the cysts to release their eggs. All system functions are controlled and operated by a touchscreen interface software. The performance of the robotic instrument is evaluated using soil samples infested with SCN from two farms at different locations and results were comparable to the conventional technique. Our new technology brings the benefits of automation to SCN soil diagnostics, a step towards long-term integrated pest management of this serious soybean pest.

Despite being the single largest cassava-producing country, yields in Nigeria remain consistently poor and among the lowest. Regionally, yields are also particularly low across Africa. Pests and pathogens, including plant-parasitic nematodes, play an important role in this current yield deficit. African countries are not only faced with the problem of food security but also that of nutritional deficiency, due to limited micronutrients in the diet. In this study, six biofortified cultivars were evaluated for their response to inoculation with approximately 30 000 root-knot nematode (Meloidogyne incognita) eggs in 30 l pots in Nigeria. All cassava cultivars proved highly susceptible to M. incognita infection after 6 months, with nematode reproduction factor ranging from 7.0 to 44.8. Galling was common on feeder roots and gall index scores were recorded between 4 to 5 (on a scale of 1-5 where 5 100 galls). Meloidogyne incognita infection significantly reduced plant height, stem girth, fresh plant mass, fresh storage root number and storage root weight. Percentage yield loss of between 41.8-88.4% was recorded in M. incognita-infected plants compared with non-infected controls. Although M. incognita reduced storage root weight, it did not necessarily affect the nutritional quality (total carotenoid) or dry weight percentage of the biofortified cassava cultivars. Total carotenoid and dry weight contents of the control cultivar were similar to some of the biofortified cultivars. The high susceptibility of the biofortified cassava cultivars to M. incognita infection indicates that substantial yield losses are likely being experienced by farmers, as this nematode pest is prevalent across sub-Saharan Africa and the tropics.

The following aqueous leaf extracts; Sinopis arvensis, Sisymbrium irio, Solanum nigrum, Chenopodium album and Portulace oleraceae alone and in combination with the nematophagus fungus, Hirsutella rhossiliensis were evaluated against the reniform nematode, Rotylenchulus reniformis infecting tomato cv. Strain-B3 under greenhouse conditions. Results indicated that most treatments tested obviously caused remarkable increase in tomato plant growth with various degrees. The application of the fungus + S. irio together revealed the highest value of percentage increase of the whole plant fresh weight as well as shoot dry weight which was amounted to 54.15 %, 114.81 % followed by S. irio alone (53.75 %, 107.41 %), respectively. All treatments showed significant reduction in total number of the reniform nematode in soil and roots of tomato. Higher reduction in the reniform nematode population density was achieved by the application of S. irio integrated with H. rhossiliensis (87.71 %) followed by S. arvensis plus H. rhossiliensis (85.89 %), S. irio (82.92 %) and then P. oleraceae combined with H. rhossiliensis (82.49 %). However, significant reduction in egg-masses numbers was achieved from all treatments in relation to nematode or wheat grains alone.

Deplorable soil fertility characterising resource-poor agrarian communities remain a major constraint for sustainable global food security. Re-mineralisation through application of rock dust encourages maintainable productivity of degraded soils. This
study was planned to appraise the effect of rock dust on root-knot disease of Citrullus lanatus caused by Meloidogyne incognita (Mi). The experiment was laid
out in Randomised Complete Block Design with three replications. Soil and root populations of the nematode were estimated and galling damage of the crop was
diagnosed and censured at 60 days after inoculation. Additional data were collected on vine length, number of leaves, days to 50% flowering and dry matter content.
Data collected were subjected to analysis of variance and the means were separated using least significant difference. The treated watermelon plants were low in rootknot disease, significantly reduced Mi nematode populations and flowered far much earlier ( p = 0.05) than the untreated organic watermelon plants.
Keywords: Citrullus lanatus; Meloidogyne incognita; non-chemical control; organic
farming

We now know, of course, that microorganisms have always played an important, if not essential role, in the biosphere with fermented foods and beverages, nutrient cycling, plant and animal diseases and agriculture foremost in their sphere of activities. Within the last twenty years, microbiology has received two enormous boosts with the developments in microbial genetics and genetic engineering probably being the most influential and the greater awareness of pollution and environmental sustainability following a close second. Biological control of plant pests including nematodes has still a long way to go and one cannot negate its efficiency in making a significant contribution to the integrated management programmes and strategies against phytoparasitic nematodes. Robust and durable system of natural suppression are almost certainly operating in some fields, but they need to be sought out and contribution factors should be identified, so that farming system can be modified to enhance suppressive mechanisms. Understanding microbial diversity and the multitrophic interactions that are manifested in the rhizosphere will play an important role in managing plant-parasitic pest nematodes.

Surveys were conducted in Rabia Road nursery, Daniyal Nursery, Noorani chowk and Gujrabad nursery, Khuzdar; Umrani nursery, Wadh, Kalat Town and Surab nursery during February and March 2013 and 2014 to study the occurrence and control of nematodes associated with pomegranate nurseries in Balochistan, Pakistan. Among them Helicotylenchus digonicus was the most frequently observed nematode. Different treatments were used to control the nematode associated with pomegranate seedlings and the results obtained were subjected to factorial analysis of variance (ANOVA), followed by least significant difference (LSD) at p < 0.05.

Kabupaten Cianjur merupakan salah satu sentra produksi tomat di Jawa Barat. Pada tahun 2011-2012 luas lahan tomat di Jawa Barat meningkat 6,31%, namun produksi menurun 17,14%. Survei ini bertujuan untuk mengetahui keragaman dan kelimpahan nematoda pada tanaman tomat serta pengaruhnya terhadap pertumbuhan tanaman tomat. Contoh tanah diambil dari dua petak lahan yang diperlakukan dengan sistem tanam monokultur dan polikultur. Survei dilaksanakan di Desa Pasir Kampung Kabupaten Cianjur - Jawa Barat, dan analisis di Laboratorium Nematologi Departemen Proteksi Tanaman IPB. Setiap petak tanah diambil 10 contoh tanah. Variabel yang diamati pada survei adalah: genus nematoda, kerapatan dan penyebaran nematoda, frekuensi penyebaran nematoda, nilai dominansi nematoda serta pengaruh kerapatan nematoda parasit tanaman terhadap pertumbuhan akar dan tinggi tanaman tomat. Hasil survei menunjukkan bahwa pada setiap contoh tanah mengandung nematoda parasit. Terdapat tujuh genus nematoda parasit pada kedua lahan, yaitu Meloidogyne spp., Pratylenchus sp., Rotylenchus sp., Ditylenchus sp., Helicotylenchus sp., Hoplolaimus sp., dan Xiphinema sp. Nematoda dari genus Meloidogyne spp. memiliki kerapatan tertinggi dibandingkan dengan nematoda lain. Pada lahan monokultur penyebaran Meloidogyne spp., Pratylenchus sp., Rotylenchus sp., Ditylenchus sp. mencapai 100%, sedangkan pada lahan polikultur nematoda yang memiliki penyebaran hingga 100% adalah Meloidogyne spp., Pratylenchus sp., Rotylenchus sp. Kerapatan tujuh nematoda tersebut memiliki pengaruh terhadap tingkat kerusakan akar dan tinggi tanaman tomat. Semakin melimpah jumlah nematoda parasit tanaman, tingkat kerusakan akar semakin tinggi. Semakin rendah jumlah nematoda parasit tanaman, tanaman tomat berkembang lebih tinggi.

Kata kunci: monokultur, nematoda parasit tanaman, polikultur, tomat

The soybean cyst nematode (SCN), Heterodera glycines, is the most damaging pathogen of soybeans in the United States. To assess the severity of nematode infestations in the field, SCN egg population densities are determined. Cysts (dead females) of the nematode must be extracted from soil samples and then ground to extract the eggs within. Sucrose centrifugation commonly is used to separate debris from suspensions of extracted nematode eggs. We present a method using OptiPrep as a density gradient medium with improved separation and recovery of extracted eggs compared to the sucrose centrifugation technique. Also, computerized methods were developed to automate the identification and counting of nematode eggs from the processed samples. In one approach, a high-resolution scanner was used to take static images of extracted eggs and debris on filter papers, and a deep learning network was trained to identify and count the eggs among the debris. In the second approach, a lensless imaging setup was developed using off-the-shelf components, and the processed egg samples were passed through a microfluidic flow chip made from double-sided adhesive tape. Holographic videos were recorded of the passing eggs and debris, and the videos were reconstructed and processed by custom software program to obtain egg counts. The performance of the software programs for egg counting was characterized with SCN-infested soil collected from two farms, and the results using these methods were compared with those obtained through manual counting.