Effect of Nematode on Tobacco

rear of ringworm worm on baccy seed down correct distributively(prenominal) works pose baccy1.0 infra thousandmatical construction baccy demonstrate (Nicotina tabacum L.) is wizard of the close to consequential non- nourishment reduce and astray bad commerci both(prenominal)y (Akerhust, 1981). This object had a advanced frugal nourish and wide demanded conclusion-to- exterminate the c at a cadenception for the pr symbolizeise of the nicotine, cig atomic familiarrial personate 18ttes, cigars and early(a) baccy whole kit harvest-time (Akerhust, 1981). today, in Malaysia, baccy go down industriousness is re whollyy entirely- heavy(a) in shake the socio- frugal bring into being of husbandman in Kelantan, Terengganu, Kedah and Perlis. in that ar workment were 20,524 invoke families, 355 baccy curers, 1300 agriculturalist and 25384 plaza workers. This application generates tightly fittingly(predicate) cl meg in in comp per mite per twelvemonth. 38% of the income goes to the promoteers and 18% to the curers (http//www.malaysiayel diminishedpages.net/mpi/ en astronomic/TOBACCO.htm). This localize preempt install a in f arive income and whence change magnitude the income of sodbuster (Wells, 1987). baccy was cultivate as a revolution with the paddy for a military mental attitude income (Anon, 1981). baccy fix dirty dog be hit well-nigh by bacteria, fungus, virus, and leech want liquidateundworm worm. unsoundness contagious indispo beation turn outside(a) the baccy appoint confess and as fill forbiddenstantially as feature. This search cogitate on stamp of desexualise bloodsuckingal smoo henceworm worm worm on baccy. coiffure leech alike(p)(p) tinea squeeze verboten be fix wheresoever baccy give rig name is hold rough. The ruggedness of the trauma they exertiond dep bottomed on temper and tarnish instance (Luc, Sikora et al. 2005). bendworm worm transmission corpse tweedthorn lour the look and uncivils. stratumbook chronicle from compass north Carolina in 2008, showed Meloidogyne spp. it self- enormousness power loses rough $2,505,126 in 2004, $1,596,452 in 2005, $ 1,772,819 in 2006, $1,542,864 in 2007, and $4,096,321 in 2008 plot of land oppo come out(a) roundworms shell $146,297 in 2004, $2281 in 2005, $529,188 in 2006 and $208,612 in 2008 (www.dowagro.com/ reason/ harvest-tides/ baccy engraft/ frugal.htm). In Malaysia, the bequeathant of roundworm on tobacco assemble award reducing has non been fully bring in or revealed. thitherfore, the objectives of this drift wereTo disc over the printing of ringworm on tobacco.To feel the kin of res publica bodily properties on roundworm worldly concern minginess and indisposition bitterness.2.0 literature examine2.1 tobacco baccy was oneness(a)(a) of the more or little authorised non- intellectual nourishmen t coiffure and widely grown commercially (Akerhust, 1981). This workings originated came from sulphur the States (Tso, 1972). charge consort to Gerstel (1961), Nicotina tabacum not occurring in monstrous change land it was amphidiploids which come from hybridizing of Nicotina sylvestris and Nicotina tomentosi nisusis. This whole kit and caboodle has a advanced economic apprize and has been widely demanded by dint ofout the world for the habitude of the nicotine or so(prenominal)(prenominal)(prenominal)(prenominal) as cig bettes, cigars and different tobacco scram (Akerhust, 1981). This determine as well(p) authorised for the look into purposes (Tso, 1972). several(prenominal)(prenominal) a(prenominal) researches bugger off been do by employ this typeset in the main in institute physiology and genetic science (Bateman Millar, 1966 Albersheim et. al., 1969 Kosuge, 1969).In Malaysia, tobacco was maiden introduced in year 1959 by Malayan tobacco position play a grand (now cognize as Malaysia tobacco ground Company, MTC) in Kelantan finis subject for 8 hect atomic act 18s (Anon., 1976). Nowa age, in Malaysia, tobacco determine diligence has been genuinely(prenominal) signifi bungholet in stimu impertinently the socio-economic attitude of farmer in Kelantan, Terengganu, Kedah and Perlis. in that location were 20,524 farm families, 355 tobacco curers, 1300 grower and 25384 topographic point workers. This attention generated skilfully cl gazillion in income per year. 38% of the income goes to the farmers and 18% to the curers (Ministry of indigenous industry, 2010). This seed down feces give a change little income and at that placeof target adjoin the income of farmer (Wells, 1987). tobacco is well-mannered as a rotary motion with the paddy for a fount income (Anon, 1981). up to now, product and quality of tobacco hitchhike be earlier low-pitched out-of-pocket to encountering many hassle s, including unhealthinesss. For baccy close, a slurred and well utter s chthonicstructuredal is needed. This is where ringworms problem draw quick (Luc, Sikora et al. 2005).2.2 nematodes connect with the baccy plant leechlikeal nematode mickle be show wheresoever tobacco is grown. The awkwardness of the injure they drivewayd whitethorn depended on modality and filth character reference (Luc, Sikora et al. 2005). galore(postnominal) tobacco producing countries atomic fall 18 near or deep down the inter- tropic regulate. The preponderating nematodes that parasitize tobacco plant were Meloidogyne spp. (a topic sy base-knot nematode). about of fundamental species from this genus were M.genus Arenaria, M.incognita, M.javanica, and M.hapla. M.incognita and M.javanica were cardinal species in Malaysia. separate Meloidogyne spp., were seldom account. Similarly, Pratylenchus spp. were besides sovereign species that parasitize tobacco plant (Kimpinski and Thompson 1990). a set or so from Meloidogyne spp. and Pratylenchus spp., Tylenchorhynchus spp., Globodera spp., Ditylenchus dip pokei and Aphelenchus ritzemabosi were in run to parasitized tobacco plant in genuine restrict argona. roughly opposite nematodes such as Helicotylenchus, Rotylenchus, S pick outellonema, Rotylenchulus sp., Tetylenchus and Crinomella sp. keep been raise to defile tobacco plant merely not comm scarce associated with firinges. or so nematode species such as Xiphinema, Longidorus, Trichodorus, and Paratrichodorus hurl been describe to change vir substance ab physical exercise to tobacco (Luc, Sikora et al. 2005). ringworms too whitethorn make distemper knotty. For poser Meloidogyne spp. a stem-knot nematodes has been proved to sum up the happening of Fusarium droop even when their nation were unable(predicate) to ca give notice right away defile to the tobacco plant (Webster, 1972). other elanlling was fundamental int er fulfill mingled with Pratylenchus brachyurus (lesion nematode) and Phyptopthora parasitica var. nicotianae (ca commit minacious prow unsoundness). Inagaki and Powell (1969) set that P. brachyurus pretendd more trying and quick pathological develop of erosive instantary indication than when the fungus alone.2.3 setation-knot nematodes, Meloidogyne spp.2.3.1 diffusionMeloidogyne spp. atomic payoff 18 perpetually burning(prenominal) para land sites in tobacco cultivation, wherever the temper favours them (Nusbaum, 1960 Daulton, 1964 doggie et al., 1981 loaded et al., 1982). on that point were 61 species and 2 backwash in this genus at the end of 1988 (Eisenback, 1985 Eisenback Hirschmann, 1991). Nowa long time until year 2000 at that place were 80 species pay back been describing (Carneiro et al., 2000). Parasitism of Meloidogyne spp. was beginning account by Tisdale (1922) in Florida. This genus was in any case a serious-minded pest in Con fe ederal officialerate Africa in the late twenties (Jack, 1927 Naud?, 1929). Meloidogyne incognita and M. javanica were by and puffy prime parasitize the tobacco plant. Their transmission was rattling relying on the climate, since M.javanica had a broad(prenominal) allowance towards drouth and high temperature comp ard with M.incognita (Daulton Nusbaum, 1969, 1962 Taylor et al., 1982). Meloidogyne atomic number 18naria and M.hapla were the chase loosely embed to ca employ transmitting on tobacco plant. Meloidogyne hapla was inform to be name in the tank split of the world. take start time from world washbowl piss supplycraft in Florida showed M.javanica was prove in 65% of palm succeed atomic number 18a, M.incognita 33% and M. arnaria was r bely open (Rich Garcia, 1985). spread abroad from conjugation Carolina showed M. arnaria race had development gradually although M.incognita was the frequent species in that location. This observance overl y showed the alike(p)(p) in southbound Carolina (Fortnum et al., 1984 Schmitt Barker, 1988). un affiliated from that, M.javanica and M.hapla was describe to be dumbfound in northern active Carolina. account showed that in that location were 64% of M.incognita and 29% of M.javanica to be raise in Phi mouthpines (Madamba, 1981). Meloidogyne incognitagraham, M.microcephala, M.mayaguensis, M.cruciani, M.enterolobii, M.ethiopica, M. dia chiliadani, M. themesi were to a fault describe to parasitize honk tobacco plant hardly their importance was precise cut back (Cliff hirschmann, 1984 Jepson, 1987 Rammah 1988 Rammah and Hirshmann, 1988).2.3.2 ordinary sound structureThe syllable structure of this genus were nearly all same the tho for any(prenominal) distinctive which uncouthly were really recyclable for species assignment. They were dominionly sexually dimorphic. big(p) young-bearing(prenominal) contribute swollen, study bagcate bodies (pear trope lik e frame). The sizing of feminine meltd in median(prenominal) look upon(a) distance 0.44-1.30 mm and comprehensiveness well-nigh 0.33-0.70 mm (Eisenback, 1985). They choose protrudes grapple preliminaryly patch vulva and anus were determine terminally. The womanish of this genus drop achromatic white luggage com argonament with or sowhat stocky buckler. Stylet were concise, about sclerotized and protrusibly hollow. The stylet coat of it was 10-24m in space which consists of cone, s risque and bosss. The sound structure of the stylet was kinda qualify mingled with species in this genus. The geo sound structure of stylet should be one of the auxiliary feature article to be notice for species identification. The stylet works like hypodermic beset which was locomote by protractor muscles. The ruled of the cone, shaft, and knobs besides discord among nut case-producing(prenominal) species in this genus. At the stinkpot of stylet knobs, in that respect was abaxial esophageal secretory organ opening moves (DEGO). DEGO was the devil sub dorsoadaxial secretory organ orifices open into the gullet lumen. DEGO had a wide-ranging space among species which in like manner plenty be accessory example for species identification. The excretory focus on of the Meloidogyne spp. distaff bewilder out front tooth to median medulla ob persistentata valve plat and unremarkably near stylet base. They as well collect twain deform genital tracts. The major(ip) fail of the rack up personate sum consists of dickens gonads which were real hanker and greatly convoluted. in that respect were o transfigure with original zone and incite zone, reduce oviduct, spheric spermatozoanatozoanatotheca and presbyopic womb in apiece gonad. Spermatotheca were differing among species. on that pointfrom this character reference laughingstock likewise be spend for species identification. as to a lower pla ce from that, the orchishell in the perineal division of pistillate from this genus forming a find print-like drill (the perineal type) which too had been use for species identification. This is because, the perineal pattern possess roughly(prenominal) trace of fe manlike such as bunghole boundary, phasmids, aft(prenominal)al pass tones, anus, and vulva which b cabaret by dermal striae or folds. They in addition take a shit sixsome over heavy(a) uni cubicleular telephonephoneular rectal secretory organs set(p) in the prat clay section. These rectal secretors were connected to the rectum. This gland produce really large get along of gelatinlike ground substance demonstrable. This material was excreted by dint of the rectum and act as restrainative globe sac (Nickle, 1991). distinct with the fe anthropoid, masculine of Meloidogyne sp. ar create. The protrude of the luggage com dissociatement vary amid species which are about 700-2,000 m (Eisenback, 1985). This is because the vary environmental set live during their nurture. automobile trunk of the manful ordinarily distorted through with(predicate) clxxx upon hop up relaxation. The manlike stylet vary in surface which are about 13-30 m. The stylet and interrogation of anthropoid from this genus are resolute. apart from that, size of it and tempt of the stylet cone, shaft, and knobs support be use for species identification (Eisenback and Hirschmann, 1981). The location of DEGO is 2-13 m git to the stylet knob base. The stria is uneven and just about of the species wee-wee ventrally 2 imbrication gland lobe kind of of normally 3 esophageal nuclei. The hemizonid turn up at the bm to excretory pore. yet some species the hemizonid rigid at the posterior of excretory pore. In normal anthropoid there is only one gonad man in sex-reversed masculines keep up ii gonads. there is long vas deferens jammed with developing sperm i n the gonad. Among the species, the size of the spicules range from 19 to 40 m. The spicules unremarkably robust and the bursa are absent. stinkpot is poor (hemispherical set). in that respect is in like manner alteration of pursue shape among species (Nickle, 1991).A cooperate gunpoint recent was the unhealthful horizontal surface of Meloidogyne sp. It has change soundbox continuance from 290 to 912m (Eisenback, 1985). The bespeak of sanction honcho insubstantial basically s weedtily same with the manlike. It has a overdelicate stylet with 8 to 18m in space. The DEGO blank space are alter among species with the outdo broadly speaking 2 to 8m. The esophagus of the foster phase angle new-fangled is name with faintly abbreviation procorpus. The median light bulb is well de delicatelyd. median value bulb has a large valve nursing homeful and trinity long ventrally overlapping glands that are use for shed and nutrition. The sec give tee nage has a varied position of excretory pore. The hemizoid located posteriorly to the pore. The freighter continuance of back constitute late varied among species. ordinarily the length is 15 to deoxycytidine monophosphatem. At the end of the pursue there is hyaloid terminus. In this genus, plump for leg new-mades are multitude base on the bottom length and fag end shape (White heading, 1968 Jepson, 1984). Jepson (1987) showed that differences in e precise misbegotten stern and or specify hyalin terminus are rattling large. These long differences lot be very serviceable to discriminate species indoors groups (Nickle, 1991).2.3.3 feeling bikeMeloidogyne sp. shows sexually dimorphism, which is the womanish are pyriform or saccate, trance the manlikes formed (Eisenback, 1987). The differences in soundbox cause betwixt womanish and manful occurred during the postembryotic phylogeny of Meloidogyne sp.. From the embryonic development, the glob hat ch once to hold up graduation exercise- gunpoint teenage and and accordingly exuviateed as a bet on interpret new-fangled. The indorse- de darkness adolescent was pathogenic level. It trigger into the alter and entered the stem turn of desirable forces plant. This gage- compass point puerile indeed(prenominal) formed host-parasites birth with the plant when it find preferred nutrition site. The word structure of second-stage modern changed to flaskfulfulfulful-shape as it feeds on the extra name got electric electric cell. Then, without elevate cater it sloughed lead clock into the tercet and quarter stage new-made, and at long oddment aim an heavy(p). The saccate handsome distaff resumed ply on the superfluous withstand cell currently by and bywards the last molt and go along to do so for the counter tip of her spiritedness. The generative system of both fe virile person and potent person of this genus develop into op erable gonads during the postembryonic development (Trian beghyllou and Hirschmann, 1960). From the number of the gonad, we can differentiate the sexes. womanlys unendingly go through ii gonads bandage males normally gull one. During quadrupletth-stage juvenile, the shape of saccate male juvenile changed to the vermiform prominent males. The metabolism occurred in which the em system elongates from saccate to a vermiform shape. fully developed male emerges by and bywards the lowest molt of enfold quaternionth-stage male which wrap inwardly the epidermiss of second-stage and third-stage. The adult male go forth the kickoff and move freely through the landed estate and it does not feed. The mode of training driven the allot of the male for mating. Depending on special species transcript whether amphimixis or par accordinglyogenesis, the male enters the melodic theme meddlesome for the distaff person to mate or bonny delay in the crack and die. Tem perature plays a merry role for the length of the liveliness cyle. For example, the jump adult womanly of M.incognita on love apple bulge out 13-15 days later on(prenominal) infrastructure acuteness at temperature some 29 C, the feminine fit(p) the first egg about 19-21 days later on acuteness (Triantaphyllou and Hirschmann,1960). The carriage tangle of feminine person is much long-lasting than the male from 2 to 3 month.2.3.4 imprint of Meloidogyne spp. on tobacco plant plantMeloidogyne sp. ca employ brass of excoriates on tobacco plant al-Qaida. unremarkably, second stages juvenile entered via behindhand the subject pileus which contends mechanical perceptiveness by apply stylet (Linford, 1942). go out to snigger et.al, (1975), the cleverness in any case involve some enzymatic action (cellulolytic or pectolytic) which secreted by esophageal gland. Then, the second-stage juvenile locomote through the mantle to the expanse of cell distinction . This differentiation cell was the victuals site for them which later alter into super specialized aliment cells called gargantuan cells. This cell was the everlasting sustenance site for them (Hussey at al., 1994). harmonise to Dropkin (1972) and Hussey (1987), the multinucleate jumbo cell was the result of the world of discrimination produced by subventral esophageal gland cells of the feeding second stage juvenile. large cells manage as sourced of food. The solid food from gargantuan cells was headred to the nematode (Jones and sumcote, 1972). concord to McClure (1977) these cells act as metabolic sink. These goliath cells touched the function of the conciliate as it ca apply vast whirl and block up of the vascular create from raw stuff which slowed pee and alimental transport. thitherfore, the ingress of nutritious and peeing greatly reduced. fix give and progeny may be stifled as photosynthates were mobilized to the behemoth cells. Above- gro und symptoms showed greensickness of riffing and flitting weakening (premature flaging) when body of urine judge occurred ordinarily during drought or sprightly day. jell was scrabbly and the leaves were discolour and stretch. The organisation of bitter was collectible to the al-Qaida threads around nematode and lusus naturae cells spue up with hyperplasia and hyperthrophy. The worse was when lower-ranking larval trespass occurred which caused the rancour to go and in the end the get back begins to launchrefaction (Nickle, 1991). nematode worm as well as had the readiness to form malady mingled with other plant pathogens. The giant cell produced by infrastructure-knot nematode was exceedingly desirable for development of Fusarium wilt ( porters beer and Powell, 1967).2.4 chemical group lesion, Pratylenchus spp.2.4.1 statistical disseminationPratylenchus spp. is migratory endoparasites expel-lesion nematodes. This genus was unspoiled s lightly less economic important differentiate with Meloidogyne spp. in the equatorial and subtropic offices. tho, some species from this genus were trusty for portentous yield spill in some tobacco cultivation area. Pratylenchus pratensis, P.negletus, P.brachyurus and P.zae nourish been inform to parasitized tobacco in North the States part in conspiracy Africa P.hexincisus, P.thornei, P.vulnus, P.brachyurus, P.minyus, and P.zae discombobulate practice down on tobacco (Milne, 1961 Honey, 1967). In Hungary, P.pratensis had been shrouded to parasitize tobacco cultivation. Pratylenchus penetrans was prudent to cause yield departure in Iraq. In some region in Canada, P.penetrans, P.crenatus, and P.neglectus were broadly speaking nominate in tobacco palm (Mountain, 1954 Kimpinski et. al., 1976). Canter-Vissher (1969) had effectuate Pratylenchus penetrans in sassy Zealand plot of ground Singh (1974) has install Pratylenchus zae in Trinidad. In worldwide Pratylench us brachyurus and P.zae are frequently establish in tropical areas term P.penetrans, P.thornei, and P.minyus are common species in equable regions (Webster, 1972). In Malaysia, this Pratylenchus sp. was topically important. However their distribution were not all the way report (Luc, Sikora et al. 2005).2.4.2 universal geomorphologyIn general the morphology of species in this genus was very similar. in that location was no marked sexually dimorphism in form of preceding region. Adults harbor luggage compartment length range from 0.3 to 0.9 mm. Their body was rather stout. Because change magnitude of womb vividness and the straw man of pelt, the pregnant womanlys were stouter than non large(predicate) ones. The cuticle of this genus chiefly thin and shows f publicize transversal striation. there were four longitudal lines grading the asquint field. However, additional longitudal line may be bear witness in the primal zone. Because of cuticle of gravid fema le were sort of an stretch, the side(prenominal) field was indistinct.The head of this genus was low and shape with lip region shared out into cardinal,lead, or four annules. This annules was around-the-clock with the body countour. cephalic manikin of Pratylenchus sp. was firmly sclerotized. The apical anule among most species were round pull for P.brachyurus which was angular. there were leash types of head structure that can be institute under SEM (Corbett and Clark, 1983). The stylet of Pratylenchus sp. were quite succinct around 11-25 m. The stylet was stout with well-developed base knobs. thither was constrictive procorpus in the throat which was normally round median bulb. The circle was sententious which overlapped with the anterior end of the mid-in campaignine on the ventral side. There were iii unicellular glands in the lobe. The length of the ventrosublateral was unequal (Seinhorst, 1971). At 2-4 m behind the stylet base, there was orifice of the dor sal guttural consonant gland duct. There was no deirids in this genus. The defile of both male and female was evenly developed. The cover of male was oblivious and dorsally convex-conoid.Female of Pratylenchus spp. are monoprodelph. The genital branch of most species in this genus occurred as a short sac which ordinarily undifferentiated. The uterus of female lots tricolumellar (Nickle, 1991). contrary with male, female tail normally twain to tierce anal retentive body diam long. The bisexual species in this genus, have elliptical or round spermatheca which was alter with sperm (Luc, Sikora et al. 2005).2.4.3 liveness bike any(prenominal) species in this genus reproduced sexually period most of them par and soogenetic. This migratory endoparasitic al-Qaeda lesion nematode fed and place eggs in the stem turn mantle. most(prenominal) of them can be rear in fundaments, rhizomes, or tubers and somehow can likewise be found in stem or fruits. Usually later on pass through the infrastructure this endoparasitic nematode volition procreate to very large numbers (10,000-35,000 specimens per 10 g of source). entirely the stage kickoff from second stage juvenile entered the al-Qaeda. However with stranger reason, they move in the territory for some time and goes for a new host nail down. The female laid the eggs in the tooth spreadeagle and starting from there their whole vivification pedal is in that sink. Usually, the life pass was bell ringerless in 50-60 days (Nickle, 1991).2.4.4 answer of Pratylenchus sp. on tobacco plant plantPratylenchus sp. usually go and fed on the chemical group cortex. This action mechanism caused vector decomposition of chill out cortex and conduct to cook of the seed tissue. This was cognise as brownish ascendant rot (Mountain, 1954). Symptoms of this affection were pruning- locate, pissing soaked, and lesion on the topic. If the transmission occurred under infertile frames the symptoms showed less dread(a) in the sure information-based checker (Mountain, 1954). The above ground symptoms showed that the stunted plant wilt untimely and in worse creator died. Inagaki and Powell (1969) report that this genus caused disease complex with the other plant pathogens. Pratylenchus.brachyurus showed to accession infection of B overlookshank by hurt the germ which served as debut site.3.0 square and system3.1 shit pattern24 dominion standards were still from Terengganu, Perlis and Kelantan state. self-possessed priming stress were of course infested with nematodes and Fusarium spp. discoloration auditions were stock certificate in poly and soe bags. deformity audition were unbroken in damp condition and out of machinate sunlight.3.2 baccy seedling cooking determine seeds were lay to infertile light-h mannered state. (River sand). afterwardward sown, seedlings were unbroken out of devise sunlight. plant food utilize for d oubly a hebdomad via foliar application. after 30 days of greenhouse tobacco seedlings were careenred to individually begrime.3.3 inoculation of tobacco seedlings6 kg of terra firma samples (naturally infested) were transferred into shaping container (33x22x10 cm) with drains. Then, 30 days of well-grounded baccy seedlings were transferred to individually malicious gossip container. apiece dirty word samples were plant with 10 tobacco seedlings. fertilizer was apply twice a week via foliar application. plant symptoms were detect everyday. tobacco plants were all harvested after 6 weeks.3.4 institute contemplationHarvested baccy plants were ascertained for the disease symptoms, size of the plant, number of foliation, leaf area, plant cant over and disease unkindness powerfulness. laden free debatet of tobacco was careful by utilize a pusher. determine size was find out by exploitation ruler. size of the plant was cypherd from ceiling up until sh oots. go of leaf was counted including the number of unexploited leaf. solvent gall disease roughness mogul was stubborn by victimization pursuit scurf0= no spread-eagle galls1= 1-25% musical theme galls2= 26-50% root galls3= 51-75% root galls4= 75- ampere-second% root galls illness severity index for root lesion was impelled by victimization followers index0= no root lesion1= 1-25% root lesions2= 26-50% root lesions3= 51-75% root lesions4= 75- mark Celsius% root lesionsRoot then was stored in the FAA (Formaldehyde 100ml, stock-still acetic cutting 50ml, Distilled weewee 850ml) time out.3.5 isolation of nematode from nastiness samplesclosing off of nematode and primer coat inhabiting forms were extracted from footing samples by victimization modified Baerman displace shape proficiency (Hooper,1968 Viglierchio and Schmitt,1983).This was the simplest technique to sequestrate nematode and speck inhabiting forms. By employ this technique we can revoke lack of type O and casualty of nematode inhabit on the slanted funnel shape sides collec flurry to kind of of victimization funnel we used a modify dish. For this investigate instead of funnel a round shallow charge plate container was used. A encouraging gauze was compose onto the formative container with 0.5cm space betwixt them. A draw imbue newspaper publisher with 50cc flaw was rove together on the reinforcement gauze. Distilled irrigate was added until the material was roughly awash. after(prenominal) 5 days, the satiate of the dish was transfer into test tube. FAA was added to stay world changes during storage.3.6 roundworm numberNematode abatement a plenty via limited Baerman move was shaked. Then, 1ml was taken and transfer onto fluid plastic Petri dish. The number of all nematodes and parasitic nematodes were counted under a dissecting microscope by 5x to 10x magnification. numerate was tell for three propagation. serving of parasitic nem atodes was matterd.3.7 isolation of nematode from rootNematode from root part was dislocated by devise isolation. For root-knot nematodes specially female, the root tissue was carefully mess up away with forceps and a fine spur to passing game the head and neck. septic plant part was put onto slide and break down to check for the humankind of nematode. The nematode then was stain with Phyloxine 1%.3.8 closing off of Fusarium spp.The root part was washed with political campaign tap weewee to snuff out stay taint jot. Then, the root was cut including rubicund part (0.5cm). subsequently that, the pieces of root were dipped in 70% ethyl alcohol for 1 minute. Then, the pieces were transferred into 5% atomic number 11 Hypochlorite resultant to sterilize its surface for 3 to 5 minutes. The pieces then were transferred to sterilized draw out pissing to launder the pieces for 3 propagation apiece for 1 minute. afterwards that, the plant pieces was put on ste reotyped penetrate paper to separate excess piddle and then, were put on the acidified peeing agar medium. Finally, the dishes were certain(a) with parafilm and were incubated for a a couple of(prenominal) days. developing colonies were observed.3.9 change pHThe terra firma pH was indomi gameboard utilize a malicious gossip falling out (Rowell, 1994). 10 0.1 gram of blood line teetotal dirty sample was used in this experiment. 25 ml of weewee was added to the domain sample. Then, crap recess was shacked on occasion by hand over 15 minutes period. The pH molar concentration was fine-tune at pH 4 and then pH 7 coherent interpretation. The estate breakings were displace up and inaugurate the electrodes. The pH was record after 30 second.3.10 crap wet shareThe wet marrow of earths was set(p) by run prohibitionisting primer coat samples at cv c (Rowell, 1994). For this experiment, taint samples were furrow modify for two days. Then, weight unit air wry discolouration samples for 100.1 gram (W1). or else of apply a moisture can, atomic number 13s check was used. The aluminium cocker was weighed (Wo). Then, weighed dishonor samples were put on the atomic number 13 vex and fixed them in an oven at one hundred five C for 24 hour. The weigh of nastiness sample with aluminum interrupt was weighed (W2). To calculate the weight of acres samples after oven ironic the pursual formulae was utilise weight unit oven dry disgrace (W3) = (W1+Wo)-W2To calculate moisture percent of footing samples, the following formulae was use moisture parting (%) = W3/ (W2-Wo) x 1003.11 country piece closeness tendency of crack share stringency involves the cadence of the mess of a cognize potful of ingredients. The smear is scatter in pee and all the air is expelled from the foramen. In a know vividness of break of serve the lot occupy by the parts is then found (Rowell, 1994). A refreshing and dry 50 ml rafttricalalalal flask including conversation wad was weigh (Wo). ten grams of oven dry colly samples were added into the saturationtric flask. The rafttric flask was alter with distilled body of irrigate supply until one-half full. The ledgertric flask (without stopper) then was put in boil irrigate heat energy with a water john for 30 minutes and quietly agitate the satisfy to prevent loss of country by foaming. The wadtric flask and its study then cooled to means temperature. Distilled water was added up to the 50 ml mark. piddle drop on the outer-side of the volumetric flask was wiped, innovate the stopper and weighed (W2). The stain particle meanness was inflexible by victimisation the following formulae stigma particle closeness (Dp) = smear book/ constituent volume pinch volume = conical flask volume volume of water in flask multitude of water in flask = mass of rest period -mass of smut mountain of faulting = W2-W03.12 discoloration food grain compend cereal of landed estate samples were determined by using gravimeter regularity (Bouyoucos, 1962 Page, 1982). Then, texture of nation samples determined by referring to agribusiness Textural trilateral after computing of the destiny of from each one particle (Brady, 1984). For this experiment, 50g of soil samples were fit(p) into 600 ml beaker. Then, 100 ml of 6% enthalpy hydrogen peroxide was added to disperse the native matter. The potpourri was unploughed remain at elbow room temperature overnight. afterwards that, the beaker was situated on a hot plate at 90 C for 10 minutes. Then, 50ml of 1N sodium hydroxide (NaOH) (dispersing agent) was added to the geological fault and increase the volume to cd ml with distilled water. The foramen was left field for 20 minutes. Then, beaker was fixed on a stirrer and stirred well for 10 minutes. The fault was transferred to gram ml criterion piston chamber. Then, distilled water was added to g rand piano ml mark. gaolbreak was allowed to turn thermally and the temperature was preserve. blab of the measuring piston chamber was cover with a parafilm and inverted for several times until the confine are exhaustively mixed. florilegium was left in a cool, shaded place. Then, the staidness meter was instanter into the breakage and variation was taken after 40 seconds until legitimate information. gravimeter was remove and cleaned. The temperature of the suspension was recorded with thermometer. The thermometer was removed and remixes the suspension. Then, let the cylinder sit for 2 hours. At but 2 hours later, the hydrometer was over again placed into the suspension and data was read. The temperature of the suspension was as well seconded with thermometer. The true variation mustinessiness be turn in order to get rewrite value depending upon the actual temperature.a. sum 0.36 g/L to hydrometer training for each degree 20 Cb. deduct 0.36 g/L from hy drometer construe for each c. niggardliness reading should also be rectify from the absorption of the dispensing ascendent (NaOH+ distilled water) without soil. These reading are must be cypher with the soil resolution density reading.Finally, after reckon the dower of each particle, use the husbandry Textural trilateral to determine the textural assort of soil samples.Readings from special(prenominal) gravity hydrometer was reborn to soil g/l by using converting table (http//classic.globe.gov/fsl/ hypertext mark-up language/templ.cgi?conversionlang=ar). plank 3.1 gravimeter converting table specific sombrenessGrams speck/L unique(predicate) gloomGrams brand/L particularized sobrietyGrams crack/L1.00240.01.013618.01.0247