一种准确测定土壤根结线虫种群数量的方法

为探索一种准确测定土壤根结线虫种群数量的方法,采用NaOCl消解附着在土壤病残体上的根结线虫卵囊与线虫常规离心技术相结合的方法,测试了不同NaOCl浓度、离心时间对根结线虫回收效率的影响,筛选出最佳测定条件,并与浅盘法、离心法、Byrd法等线虫常规分离方法进行了比较。结果表明,在9个NaOCl处理浓度中,1.0%~2.5%NaOCl处理对土壤根结线虫的回收效率显著高于3.0%NaOCl处理,达到71.6%~99.4%。在1.0%、1.5%、2.0%和2.5%NaOCl处理30 s后,分别离心2~5、2~3、2~3 min和2 min对根结线虫的回收效率明显高于其它时间处理,在每50 m L土壤平均接种976粒线虫卵的人工病土中,上述处理分离线虫数量分别达到804.3~930.0、810.7~838.7、843.7~867.0和820.7。本研究的改良方法对自然病土中根结线虫的分离数量分别是浅盘法、离心法和Byrd法的29.3、13.4和2.1倍,显著提高了土壤中根结线虫的分离效率,可准确测定土壤根结线虫种群的数量。     

Host status effect of cowpea and sunflower on the populations ofMeloidogyne javanica andRotylenchulus reniformis

A highly susceptible cowpea,Vigna sinensis cv. Baladi plants were tested as trap plants for eitherMeloidogyne javanica orRotylenchulus reniformis under greenhouse conditions. The plants were gathered by cutting them above the surface of the soil or by uprooting them, 1/2, 1, 3, 6, 12, 24, 36 and 48 days after nematode inoculation. Both of the mentioned nematodes began to mature and lay eggs after the 12th day from their inoculation. Hence, it is advised to pull up cowpea plants from 3–12 days after nematode inoculation. After planting sunflower,Helianthus annus cv. Miak replacing cowpea, the nematode populations were higher, in most cases, on sunflower plants replacing cutting cowpea than those on sunflower replacing uprooted cowpea. The highest percentages of nematode reduction were 98.55 and 99.57 forM. javanica and 95.09 and 92.90% forR. reniformis on sunflower plants replacing cutting and uprooted cowpea plants after 12 days from nematode inoculation, respectively.M. javanica andR. reniformis decreased the length and weight of sunflower plants as affected by planting time and method of cowpea harvest. This method of nematode control is cheaper, easy and pollution free.     

玫瑰黄链霉菌片剂对根结线虫病的防治作用及对土壤微生物区系的影响

玫瑰黄链霉菌Men-myco-93-63是从马铃薯疮痂病自然衰退土壤中筛选到的对多种重要植物病原菌具有较强抑制作用的拮抗链霉菌,其发酵液对根结线虫具有较好的防治作用。为明确Men-myco-93-63活菌片剂在蔬菜根结线虫防治中的应用前景,采用温室盆栽法和稀释平板法研究了Men-myco-93-63活菌片剂对黄瓜根结线虫病的防治效果及对土壤微生物区系的影响。结果表明:Men-myco-93-63片剂对黄瓜根结线虫病具有较好的防效,1.0和1.5 g/株用量下防效分别为51.85%和55.56%,而对照药剂淡紫拟青霉颗粒剂1.5 g/株及M en-myco-93-63固体发酵物(发酵物与病土体积比为1∶200)的防效分别为54.17%和50.00%;施药60d后,该片剂对植株根际土壤中微生物区系影响较大,1.0 g/株用量下,细菌和放线菌数量分别比空白对照增加了225.45%和75.45%,真菌数量降低了66.43%,3种可培养微生物的总量比空白对照增加了195.35%,表明M en-myco-93-63片剂的施用可明显改善根际土壤中的微生物结构。     

Nematicidal activity of Ochradenus baccatus against the root‐knot nematode Meloidogyne javanica

Ochradenus baccatus is a widely distributed shrub in desert regions of the Middle East and North Africa. This plant's nematicidal activity against the root‐knot nematode Meloidogyne javanica was evaluated because it has been found to contain exceptionally high levels of glucosinolates. In in vitro assays with aqueous extracts of the plant, 100% of second‐stage juveniles were immobilized after exposure to 4% root‐core extract for 48 h; 8% root‐core extract suppressed their hatching by 87%, whereas stem, flower and root bark showed lower activity. Incorporation of root core or bark into the soil, as fresh or dry powder at 1 and 0·5% (w/w), respectively, reduced the number of nematodes recovered from the soil by 95–100%, whereas the flower and stem were much less effective. Results from further pot experiments indicated that only the root bark consistently contains nematicidal compounds which are effective in soil, whereas the nematicidal activity of the root core in soil was inconsistent. The presence of non‐volatile lipophilic and lipophobic nematicidal compounds in the root bark was suggested by extraction with different polar solvents, but these compounds do not seem to be isothiocyanates – glucosinolate‐hydrolysed compounds with nematicidal activity. Very poor host status of Ochradenus baccatus to M. javanica, M. incognita and M. hapla, but with root‐penetration rates of juveniles similar to those in tomato roots, suggest that this plant may be used as a cover plant or trap plant to reduce nematode populations in the soil.     

Accumulation and distribution of pp′-DDT and related compounds in an apple orchard. II. Residues in trees and herbage

DDT residues in or on the roots and leaves of the herbage and the roots, bark, leaves and fruit of the trees are given for an apple orchard sprayed annually (1953–1969). The distribution of DDT in both the grass and the grass roots was in circular areas of residues, with maximum values at each trunk and decreasing radially to each alley. Of the spray applied at the green cluster stage 80% was deposited on the grass sward and very little, if any, directly on the soil surface. The pp′-DDT content of the grass fell rapidly with successive mowings (from which the cuttings remained in situ) from 400 μg/g at spraying to 2 μg/g after nine months. 33 g/ha pp′-DDT was found in the herbage roots (0.87% of the total residues in the soil). The residues in the bark (87.5 g/ha) were much lower than expected after 13 years spray application. There were increased amounts of pp′-DDE, pp′-TDE and pp′-TDEE relative to pp′-DDT, indicating some breakdown on the bark, but the chief losses were attributed to volatilisation and to removal by wind and rain. The residue content of root bark varied from 3 μg/g near the emerging trunk to 0.05 μg/g at a depth of 90 cm. The pp′-DDT content of leaves at leaf fall rose from <1 ng/g after a single spring spray to 8.33 μg/g following an additional spray in late June. There was a large loss of DDT from the canopy between the June spray and leaf fall (440–480 g/ha down to 25 g/ha), attributed to volatilisation. The amount of pp′-DDT on the fruit, after a single spray, was 3 ng/g fresh weight (80.9 mg/ha out of a total of 1.0–1.5 kg/ha used).     

玉米矮化病株及其根际土壤内线虫的分离与分子鉴定

为探究玉米矮化病的病原,采用Ludox TM悬浮法对从吉林和辽宁采集的玉米矮化病株及其根际土壤和健康株根际土壤分离的线虫进行了鉴定和种群比较,并对矮化病株茎基组织PCR产物进行测序分析。结果表明:玉米健康株根际土壤分离出29属线虫,特有线虫是角咽线虫属Actinolaimus和木盾移线虫属Peltamigratus,矮化玉米病株根际土壤分离出28属线虫,特有线虫为锥线虫属Dolichorus,二者相同的线虫属有27个。矮化玉米病株根际土壤中植物线虫种类和数量明显比健康株根际土壤中高,线虫种群数量差异明显的主要是矮化线虫属Tylenchorhynchus、短体线虫属Pratylenchus和刺线虫属Belonolaimus,病株根际土壤中这3个属线虫总数约占病株土壤分离线虫总数的20.23%、11.27%和10.40%。玉米矮化病株茎基组织中短体线虫属和矮化线虫属数量占优势,2个属的线虫数量分别占测序总数的22%、14%(吉林长岭)和16%、20%(辽宁黑山)。表明玉米矮化病的发生与植物线虫相关,很可能是多种线虫共同引起的病害。     

Prevalence and diversity of plant parasitic nematodes in Northern Ireland grassland and cereals,and the influence of soils and rainfall

The prevalence and diversity of plant parasitic nematodes in Northern Ireland cereal and grassland was determined from 191 agricultural fields. A total of 18 nematode genera were detected, including economically important pests, Meloidogyne spp., Heterodera spp. and Pratylenchus spp., each of which were above economic damage thresholds in a significant proportion of the sites (92.4%, 70% and 28.6%, respectively). The detection of the root knot nematode, Meloidogyne minor (6% prevalence), was significant given its recent emergence across the turf grass sector and the prospect of M. minor becoming a common agricultural pest. Analyses of nematode prevalence and abundance highlighted significant associations with grass and cereals, soil types, soil grade (proxy for soil quality) and rainfall levels. Specifically, nematode populations varied between the two major soils (brown earths and gleys), while significant trends for increased nematode diversity and greater prevalence of both Meloidogyne and Pratylenchus with increasing rainfall were also observed. Multivariate analyses were performed to determine interactive effects and the relative importance of the factors affecting nematode populations. Notably, rainfall, in combination with either crop type or soil grade, had a significant effect on nematode abundance and diversity. The findings suggest significant changes in nematode populations have occurred over the last several decades and the possibility that these are linked to changing climate and cropping practices are discussed, as well as future concerns for plant parasitic nematode management.     

Nematodes of food legumes in the Mediterranean Basin1

Several endoparasitic nematodes have been reported on leguminous plants in the Mediterranean area. The most widespread are the root-lesion nematodes Pratylenchus mediterraneus, P. penetrans and P. thornei. Symptoms induced by these nematodes usually are not very impressive, but 50% yield loss of chickpea may occur. Among root-knot nematodes, Meloidogyne artiellia is associated with severe yield losses of chickpea in Italy, Spain and especially Syria. Tolerance limits of 0.14 and 0.02 of this nematode per ml soil are reported for winter and spring-sown chickpea, respectively. Meloidogyne incognita and M. javanica can be noxious to French bean and cowpea in sandy soil. The cyst nematode Heterodera goettingiana reduces yields of pea, broad bean and vetch when its population densities exceed 0.5, 1, and 2.1 eggs per g of soil, respectively. Heterodera ciceri occurs in northern Syria and Turkey and is responsible for economic yield losses of chickpea and lentil in fields infested with more than 1 or 2.5 eggs per g of soil, respectively. Pea and grass pea also suffer from infestation of this nematode. The stem and bulb nematode Ditylenchus dipsaci causes severe decline of broad bean, pea and probably lentil during wet seasons. Other nematodes, although present in moderate numbers, appear to have little importance.     

Potential of Heterorhabditis zealandica (Rhabditida: Heterorhabditidae) for the control of codling moth,Cydia pomonella (Lepidoptera: Tortricidae) in semi-field trials under South African conditions

Infection of codling moth, Cydia pomonella (L.), larvae by the entomopathogenic nematode, Heterorhabditis zealandica, was studied under semi-field conditions. Late-instar diapausing codling moth larvae, coccooned in perforated cardboard strips, were used in all trials to evaluate the nematode treatments involved. A morning application of 0.5, 1.0 and 1.5 MM infective juveniles (IJs) per tree resulted in 85%, 95% and 100% codling moth larval infection, respectively. Contrasting results were obtained for an evening application, with low levels of codling moth infection (<50%). Nematode treatment strips placed in direct sunlight resulted in almost no infection (<10%), opposed to strips attaining 67% infection placed in the shade.     

Direct molecular detection of the pinewood nematode, <Emphasis Type="Italic">Bursaphelenchus xylophilus</Emphasis>, from pine wood,bark and insect vector

The pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease. The international economic impact of the introduction of the PWN into new areas has highlighted the need for the development of accurate and reliable detection methods of B. xylophilus, which are essential to define aspects of its control and management. In the present study, a methodology was developed for the direct detection of PWN by conventional PCR assay, with a species specific set of primers based on PWN satellite DNA, using total DNA extracted directly from maritime pine, Pinus pinaster, wood and bark samples, and from the insect vector, Monochamus galloprovincialis. This methodology involves homogenisation of wood, bark and insects using liquid nitrogen, DNA extraction and one or two PCR amplification steps, which permit the rapid and direct detection of one single nematode present in 100 mg of wood and bark and in one entire insect without the preliminary steps of nematode extraction.     

Reaction of six Musa genotypes to root-parasitic nematodes

The reaction of six Musa genotypes to root-parasitic nematodes was evaluated under field conditions, together with the horizontal, and vertical distributions of the neomatodes within the soil profile. The numbers of Radopholus similis, Helicotylenchus spp., Meloidogyne spp. and of all nematodes in the excavated soil quadrant varied with Musa genotype. All genotypes were very susceptible to at least one nematode. Yangambi km5, Pisang Jari Buaya and FHIA-23 supported the lowest number of R. similis and Pisang Jari Buaya, the lowest number of Helicotylenchus spp. These three cultivars supported high numbers of Meloidogyne spp. Furthermore, FHIA-23 supported high numbers of Helicotylenchus spp. The horizontal and vertical distributions of R. similis in the root system of Valery, Gros Michel and FHIA-18 were very similar. By constrast, the distributions of Helicotylenchus spp., Meloidogyne spp. and the number of all nematodes varied slightly among the genotypes. Because Musa genotypes susceptible to a particular nematode showed a similar horizontal and vertical nematode distribution with populations concentrated in the vicinity of the plant base (0–30 cm horizontal distance and 30 cm depth), sampling at this site would be sufficient for the identification of the Musa plant reaction.     

Acidified peel of litchi fruits selects for postharvestPenicillium decay

Litchi fruits are fumigated after harvest with sulfur dioxide (SO₂) to prevent their rapid browning. SO₂ blocks enzymatic activity but bleaches the fruits and, if this process is followed by dipping the fruit in dilute hydrochloric acid, the appealing red color is regained. Hot water brushing (HWB) is among the alternative methods that were developed to replace the use of SO₂. HWB reduced fungal population size on the surface of the fruit peel after treatment but did not eliminate fruit infection after storage. Whereas untreated fruits were infected with a variety of fungal species,Penicillium sp. was the only fungus that developed on the pericarp after storage in fruits that had been dipped in 1.5M HCl. Fruit treated by HWB followed by handling and storage under sterile conditions suffered greater decay than fruit stored under non-sterile conditions but with more ventilation. APenicillium sp. isolated from litchi grew well in liquid medium acidified to the pH range reported for SO₂ and HCl-treated litchi fruits. Morphological analysis identified fungal isolates asP. aurantiogriseum. Internal transcribed spacer sequence analysis of five isolates suggested a sequence similarity toP. commune. Our data support the hypothesis that dipping litchi fruit in hydrochloric acid eliminates infection by common opportunistic fungi and selects forPenicillium species that tolerate low pH. http://www.phytoparasitica.org posting April 30, 2004.     

脱硫石膏对苏打盐碱土水盐入渗过程的影响

为研究脱硫石膏对苏打盐碱土水盐入渗过程的影响,2020年6月进行了一项室内土柱入渗试验,供试土壤离子组成主要为Na_2CO_3和NaHCO_3、电导率为1.89 mS·cm~(-1)、钠吸附比为10.01、碱化度为40.20%,设置了5个脱硫石膏施用水平(施用量分别为土重的0.0%,0.5%,1.0%,1.5%,2.0%),并利用Philip和Kostiakov两种入渗模型描述土壤水分入渗过程。结果表明:在入渗时间为630 min时,0.5%、1.0%、1.5%、2.0%处理累积入渗量与0.0%处理相比分别增加20.00%、43.03%、61.21%、55.15%,入渗率分别增加64.94%、233.33%、271.86%、339.83%;当湿润锋运移至60 cm时,0.5%、1.0%、1.5%、2.0%处理所用时间比0.0%处理分别减少33.33%、63.89%、63.89%、70.83%;同时,脱硫石膏显著影响了Philip和Kostiakov入渗公式参数,随脱硫石膏用量的增加而增加,吸渗率S从0.0647 cm·min~(-0.5)增加到0.0957 cm·min~(-0.5),稳定入渗率A从0.0024 cm·min~(-1)增加到0.0189 cm·min~(-1),经验指数K从0.0165增加到了0.0698;在50～60 cm土层深度随着脱硫石膏用量的增加电导率逐渐增加,土壤脱盐效率增加。综上,脱硫石膏可以加快苏打盐碱土水分入渗速率,增加脱盐效率,对苏打盐碱土改良有很好的效果。     

Designing the ideal habitat for entomopathogen use in nursery production

BACKGROUND: Greenhouse and nursery producers use entomopathogens (nematodes and fungi) to control soil pests. Although it is known that the physical and chemical properties of mineral soil significantly impact upon soil pathogens, the influence of soilless media used for plant production on entomopathogen performance is poorly understood. RESULTS: Survival and foraging distance were differently affected by sand:peat, bark and sawdust media for entomopathogenic nematodes, but not for the immobile fungus Metarhizium anisopliae. Redwood sawdust medium consistently had a negative impact upon entomopathogenic nematodes. Dividing media into individual components supported the hypothesis that redwood sawdust reduced foraging and infection abilities of S. riobrave and H. bacteriophora. Physically altering the components by adding sand significantly improved foraging and infection success for S. riobrave in media not optimum for foraging. CONCLUSION: This study is the first to highlight the importance of selecting the appropriate soilless media and pathogen species combinations to increase efficacy of biological control. H. bacteriophora was able to find hosts in a wider diversity of medium components than S. riobrave, although both nematode species performed well in peat moss and recycled plant material. These results suggest that peat moss, recycled plant material and hardwood bark are components amenable to EPN biological control programs. Copyright © 2012 Society of Chemical Industry     

Survival of Bursaphelenchus xylophilus and Monochamus galloprovincialis in pine branches and wood packaging material

Survival and development of Bursaphelenchus xylophilus (pine wood nematode) was studied for up to 40 weeks in Pinus pinaster sawn wood and branches: 30 boards (1200 × 100 × 25 mm), 30 long‐blocks (1200 × 95 × 95 mm), 10 pine branches with bark and nine branches without bark (1200 mm long). The nematode was found in all materials and through the entire sampling period, with higher abundance in the sawn wood (boards and long‐blocks). In the initial period B. xylophilus reproduced abundantly and a population peak was detected at 8–12 weeks. Subsequently, the populations declined and became dominated by third‐stage resistant larvae (J_III), and in the final sample nematode abundance was very low. Nematode decline in the wood was correlated with a decrease in the moisture content (MC) to below fibre saturation. Survival of the insect vector Monochamus galloprovincialis was also assessed in sawn boards (1200 × 100 × 25 mm, n = 31) and blocks (160 × 95 × 95 mm, n = 40). The majority of the larvae were killed when sawing the wood, although some adults successfully emerged from the boards (10% survival) and blocks (37%). These results represent a contribution to the quantification of the risks of dispersing pine wilt disease through wood packaging materials, confirming that untreated wood can support healthy and abundant B. xylophilus populations for sufficient time for vectors surviving the sawing process to complete their development, to emerge and disperse the nematode.     

长期施肥对渭北旱塬麦田土壤线虫群落特征的影响

为探究长期定位施肥对黄土高原渭北旱塬麦田土壤线虫群落的影响,以长武国家黄土高原农业生态试验站的长期定位试验（1984—2018年）为平台,调查裸地（L）以及不施肥（CK）、施氮磷肥（NP）、单施有机肥（M）、氮磷肥配施有机肥（MNP）的小麦田土壤线虫群落数量、组成结构和生态功能指数,并分析其与土壤理化性质的关系。结果表明：（1）小麦田土壤线虫数量显著高于裸地,而CK、M和MNP处理使小麦田中土壤线虫数量比NP处理提高了31.63%~56.20%;（2）施肥减缓了长期种植小麦导致的食细菌线虫（特别是头叶属）相对丰度的下降,单施有机肥的处理（M）植食性线虫相对丰度显著降低,同时,相比于CK,施用有机肥的M和MNP处理杂食/捕食线虫相对丰度分别增加了18.4%和8.24%,表明有机肥对杂食/捕食线虫的促进作用;（3）长期施肥处理土壤线虫Shannon多样性指数（H）和指示土壤健康状况的瓦斯乐卡指数（WI）分别为1.80~2.19和0.36~0.68,相较于裸地（H=2.36;WI=1.57）有所下降,但M和MNP处理的线虫成熟度指数（MI）和结构指数（SI）均高于其他处理（L、CK和NP处理）,表明施用有机肥有助于土壤食物网维持复杂的结构和成熟稳定的状态;（4）不同施肥措施下土壤全氮、有机碳、微生物生物量碳和氮以及可溶性碳的变化是影响土壤线虫数量和群落特征的重要环境因素。     

Development of a real‐time PCR method for the detection of the dagger nematodes Xiphinema index,X. diversicaudatum,X. vuittenezi and X. italiae,and for the quantification of X. index numbers

The ectoparasitic dagger nematodes Xiphinema index and Xiphinema diversicaudatum, often at low numbers in the soil, are vectors of grapevine nepoviruses, which cause huge agronomical problems for the vineyard industry. This study reports a method, based on real‐time PCR, for the specific detection of these species and of the closely related non‐vector species Xiphinema vuittenezi and Xiphinema italiae. Specific primers and TaqMan probes were designed from the ribosomal DNA internal transcribed spacer 1 (ITS1), enabling the specific detection of single individuals of each of the X. index, X. diversicaudatum, X. italiae and X. vuittenezi species whatever the nematode population. The specificity of detection and absence of false positive reaction were confirmed in samples of each species mixed with the three other Xiphinema species or mixed with nematodes representative from other genera (non‐plant‐parasitic Dorylaimida, Longidorus sp., Meloidogyne spp., Globodera spp. and Pratylenchus sp.). The method was shown to be valid for the relative quantification of X. index numbers through its use, from crude nematode extracts of soil samples, in a greenhouse assay of grapevine accessions ranging from highly susceptible to resistant. As an alternative to time‐consuming microscopic identification and counting, this real‐time PCR method will provide a fast, sensitive and reliable diagnostic and relative quantification technique for X. index nematodes extracted from fields or controlled conditions.     

Estimation of Loss in Wheat Yield Due to the Cerealcyst Nematode Heterodera avenae

Abstract

The cereal cyst nematode, Heterodera avenae Wollenweber, 1924, occurs worldwide including India where it causes ‘molya’ disease of wheat and barley. Wheat yield losses due to the nematode were estimated during 1974-76 at a few villages in Mohindergarh district, Haryana, India, by applying dichloropropane-dichloropropene mixture and dibromochloropropane singly or in combination as soil f umigants. In light soil the cereal cyst nematode infestation at 4/5 eggs and larvae/g soil approximated the economic threshold level. However, depending on the initial population density of H. avenae and other field conditions, such as soil type, moisture, soil condition, chemical application method, chemical application time etc., the increase in yield of wheat cv. Kalyan Sona ranged from 8 to 49% at nematode infestation levels of about 6 to 13 eggs and larvae/g soil when treated with these nematicides.     

生物熏蒸用于治理蔬菜根结线虫病的研究

将甘蓝(Brassica oleracea)、芥菜(B.juncea)和雪里蕻(B.junceavar.multiceps)的叶及芥菜疙瘩(B.napifor-mis)的块根组织以3.5 kg/m²的量分别施入密封于棉纱缸中的严重感染根结线虫(Meloidogyne incognita)的土壤中,在25℃和32℃下熏蒸6 d后对活体线虫的测定结果表明,在25℃下用上述4种材料熏蒸后活体线虫数量分别比对照减少80%、85%、87%和91%。32℃下的熏蒸效果明显优于25℃下的熏蒸效果,各处理土壤中均未发现活体线虫,而无处理对照土壤中的活体线虫数量为1 540条/kg土壤。对各类添加物增效作用的调查结果表明,添加麦麸的增效作用最明显。田间试验结果表明,植物材料单独或与添加物混合使用对根结线虫病的防效均显著高于太阳能和土壤还原消毒法的防效。其中芥菜添加麦麸在开花期和盛果期的防效分别为65.2%和66.0%,与对照药剂98%棉隆可湿性粉剂的防效无显著差异;各植物材料熏蒸效果大小依次为芥菜>雪里蕻>芥菜疙瘩>甘蓝。     

Development and validation of species-specific primers that provide a molecular diagnostic for virus-vector longidorid nematodes and related species in German viticulture

The nematode species Longidorus attenuatus, L. elongatus, L. macrosoma and Paralongidorus maximusare economically important pests to the viticulture industry due to their ability to vector two nepoviruses (Raspberry Ringspot Virus and Tomato Black Ring Virus) to grapevines. In Germany, these species occur in vineyard soil with other non-vector but morphologically similar longidorid species, L. helveticus, L.profundorum and L. sturhani. Species-specific primers were designed from ribosomal DNA for all seven species to facilitate taxonomic identification for non-specialists. Primers were assessed for their reliability by screening, where possible, a number of populations of each species. Furthermore, their selectivity and sensitivity were determined when challenged with closely related longidorid species and general nematode communities typical of vineyard soil. A multiplex approach using a common forward primer combined with species-specific reverse primers enabled three target nematode species to be detected in the same PCR reaction. All primers were highly specific, detecting all nematode developmental forms from disparate populations and were sufficiently sensitive to detect a single target nematode within a whole nematode community typical of a vineyard soil comprising of a range of non-target species. Given their specificity, sensitivity and reliability, these diagnostic primers should be of great benefit to both phytosanitary/quarantine services related to the viticulture industry and also as a decision management tool for growers.