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1.
In 2017, during a survey on subsistence farms and gardens in Coimbra region, Portugal, 40 infected root samples were collected and 47 root-knot nematode (RKN) isolates identified, based on esterase phenotype. The phenotypes A2, H1, Hi2/Hi4, I1/I2/I3 and J3 associated to five Meloidogyne species (M. arenaria, M. hapla, M. hispanica, M. incognita and M. javanica) were found in 43 RKN isolates. The esterase phenotype En2/En4/En5, corresponding to M. enterolobii (=M. mayaguensis), was detected in four RKN isolates from Cereus hildmannianus (Cactaceae), Lampranthus sp. (Aizoaceae), Physalis peruviana (Solanaceae) and Callistemon sp. (Myrtaceae) infected roots. In order to validate the biochemical identification of the M. enterolobii isolates, molecular studies performed with species-specific primers yielded the expected fragment of c.520 bp, and the amplification of cytochrome oxidase subunits I and II regions of 800 bp. The DNA sequences of one of the isolates were compared with available Meloidogyne species sequences in databases. The Portuguese isolate grouped with 99–100% bootstrap support with all M. enterolobii sequences included for comparison, confirming the presence of this RKN species in Portugal. In the EPPO region, M. enterolobii has been reported in France and Switzerland and intercepted in the Netherlands, Germany and the UK associated with plant material from Asia, South America and Africa. Taking into account the pathogen aggressiveness and its distribution, there is a high probability of its spread not only in the Mediterranean region but also in Europe, and of it becoming a threat to the agricultural economy, where there are no effective strategies for its control.  相似文献   

2.
Abstract

Screening tests on 162 tomato cultivars and hybrids conducted at the Division of Horticulture, Indian Agricultural Research Institute, New Delhi during 1967-8 indicated a high degree of resistance to root-knot nematodes in Nematex, VFN-8, 65N215-1, 65N255-1 and S1-120, under field and laboratory conditions. The degree of resistance varied with the species of Meloidogyne. Nematex was immune to M. javanica, M. incognita and M. arenaria. VFN-8, 65N215-1 and 65N255-1 were immune to M. incognita and M. arenaria and resistant to M. javanica. S1-120, a commercial cultivar, showed a high degree of tolerance to M. javanica, M. incognita and M. arenaria. The degree of resistance varied with nematode population density.  相似文献   

3.
Seven root-knot nematodes (RKN), including Meloidogyne exigua, M. incognita, M. paranaensis, M. enterolobii, M. arabicida, M. izalcoensis and M. arenaria are major pathogens of coffee crop in the Americas. Species-specific primers for their identification have been developed for five of them and constitute a fast and reliable method of identification. Here we report a PCR-based assay for specific detection of M. arabicida and M. izalcoensis. Random Amplified Polymorphic DNA fragments specific for these two species were converted into sequence characterized amplified region (SCAR) markers. PCR amplification using the SCAR primers produced a specific fragment of 300 bp and 670 bp for M. arabicida and M. izalcoensis, respectively, which were absent in other coffee-associated Meloidogyne spp. tested. SCAR primers also allowed successful amplification of DNA from single second-stage juveniles (J2), males and females. In addition, these primers were able to unambiguously detect the target species in nematode suspensions extracted from soil and roots samples, in different isolates of the same species or when used in multiplex PCR reactions containing mixtures of species. These results demonstrated the effectiveness of these SCAR markers and their multiplex use with those previously developed for M. exigua, M. incognita, M. paranaensis, M. enterolobii and M. arenaria constitute an essential detection tool. This diagnostic kit will contribute for specific J2 identification of the major RKN infecting coffee from field samples in the Americas.  相似文献   

4.
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5.
Several experiments were carried out to assess the performance of commercial Solanum torvum cultivars against the root knot nematodes Meloidogyne incognita and M. javanica in Spain. The response of S. torvum rootstock cultivars Brutus, Espina, Salutamu and Torpedo against M. incognita and Mi-1.2 (a)virulent M. javanica isolates was determined in pot experiments, and of ‘Brutus’ to an N-virulent isolate of M. incognita, compared with that of the eggplant S. melongena ‘Cristal’. The relationship between the initial and final population densities of M. javanica on ungrafted and grafted ‘Cristal’ onto the S. torvum ‘Brutus’ was assessed, together with the effect on dry shoot biomass. Finally, the population growth rate and the resistance level of the four S. torvum cultivars against M. incognita was assessed under plastic greenhouse conditions in two cropping seasons. All S. torvum rootstocks responded as resistant to the M. incognita isolates and from highly resistant to susceptible against M. javanica isolates. The maximum multiplication rates of M. javanica on the ungrafted or grafted eggplant were 270 and 49, respectively, and the equilibrium densities were 1318 and 2056 eggs and J2 per 100 cm3 soil, respectively. The tolerance of the ungrafted eggplant was 10.9 J2 per 100 cm3 soil, and the minimum relative dry shoot biomass was 0.76. The population growth rate of M. incognita on eggplant cv. Cristal differed from that of the S. torvum cultivars in both cropping seasons. These results suggest that S. torvum is a valuable rootstock for managing the two Meloidogyne species irrespective of the (a)virulence status.  相似文献   

6.
In 2013, during a field survey conducted in Portugal on potato, Solanum tuberosum, an unusual esterase (EST) phenotype was detected in a root‐knot nematode (RKN) from potato roots collected in Coimbra. This Portuguese isolate was purified and maintained on tomato, S. lycopersicum, and morphological, biochemical and molecular characteristics were studied. Perineal pattern morphology was highly variable, similar to Meloidogyne ethiopica and not useful for identification. The EST phenotype, from young egg‐laying females, displayed three bands similar to the Brazilian M. luci (L3) and distinct from M. ethiopica (E3). Phylogenetic analyses of mitochondrial cytochrome oxidase subunit I and the mitochondrial DNA region between COII and 16S rRNA genes revealed that the Portuguese isolate grouped with M. luci isolates close to M. ethiopica isolates. However, considering the ITS1‐5.8S‐ITS2 region, the Portuguese isolate grouped with isolates of M. luci, M. ethiopica and M. hispanica, which limits the confidence of this region for M. luci diagnosis, and its differentiation from other species with morphological similarities. The M. luci pathogenicity to potato was also assessed in 16 commercial cultivars and compared with M. chitwoodi, considered to be a quarantine RKN species by EPPO. All potato cultivars were susceptible to both Meloidogyne species with gall indices of 5 and higher reproduction factor values ranging from 12.5 to 122.3, which suggests that M. luci may constitute a potential threat to potato production. In the present study, M. luci is reported for the first time attacking potato in Portugal.  相似文献   

7.
The root-knot nematode Meloidogyne incognita is widely distributed and a major pathogen of cotton (Gossypium spp.) worldwide. The objectives of this study were to assess the genetic variability and aggressiveness of Brazilian populations of M. incognita in cotton. Five populations of M. incognita and one isolate of M. enterolobii (outgroup) were used in the molecular analysis. Our results showed that only 2.7 % of the RAPD and AFLP fragments were polymorphic. Despite the existence of two races (races 3 and 4) and two esterase phenotypes (I1 and I2), a low genetic variability among populations was observed, which might be due to the mitotic parthenogenetic mode of reproduction of this pathogen. The aggressiveness/virulence among populations towards different cotton genotypes was also studied. None of the populations was virulent to the resistant cotton genotypes M-315 RNR, TX-25, CIR1343, Wild Mexican Jack Jones and CIR1348 (reproduction factor <1). Two populations of M. incognita from the states of Mato Grosso do Sul and Parana (Umuarama) (races 4 and 3, respectively) were highly aggressive to the susceptible control FM966 and virulent to the accessions LA-887 and Clevewilt-6 that showed moderate resistance to other populations tested.  相似文献   

8.
The present research was undertaken to evaluate the effects of soil temperature on the life cycle of root-knot nematodes (RKN) on zucchini-squash in growth chambers and to assess the relationship between Meloidogyne incognita soil population densities at planting (Pi), its multiplication rate, and crop losses of zucchini in field conditions. Thermal requirements for M. incognita and M. javanica were determined by cultivating zucchini plants in pots inoculated with 200 second stage juveniles (J2) of each Meloidogyne species at constant temperatures of 17, 21, 25, and 28 °C. Number of days from nematode inoculation until appearance of egg laying females and until egg hatching were separately recorded. For life cycle completion, base temperatures (Tb) of 12?ºC and 10.8?ºC and accumulated degree-days above Tb (S) of 456 and 526, were estimated for M. incognita and M. javanica, respectively. The relationship between fruit weight and M. incognita Pi fits the Seinhorst damage function, but differed accordingly to the cropping season, spring or autumn. Tolerance limits for M. incognita on zucchini were 8.1 J2 per 250 cm3 of soil in spring and 1.5 in autumn cropping cycles, and the minimum relative yields were 0.61 in spring and 0.69 in autumn. Zucchini-squash was a poorer host for M. incognita in spring than in autumn, since maximum multiplication rates (a) and equilibrium densities (E) were lower in spring (a?=?16–96; E?=?274–484) than in autumn (a?=?270–2307; E?=?787–1227).  相似文献   

9.
Root‐knot nematodes (RKNs), Meloidogyne spp., are a major disease problem in solanaceous crops worldwide, including pepper (Capsicum spp.). Genetic control provides an economically and environmentally sustainable protection alternative to soil fumigants. In pepper, resistance to the main RKN species (M. incognita, M. javanica and M. arenaria) is conferred by the major genes (R genes) Me1, Me3 and N. However, RKNs are able to develop virulence, thus endangering the efficiency of R genes. Quantitative resistance (QR) against Meloidogyne spp. is expected to provide an alternative to R genes, or to be combined with R genes, to increase the resistance efficiency and durability in pepper. In order to explore the ability of QR to protect pepper against RKNs, five pepper inbred lines, differing in their QR level, were tested directly, or after combination with the Me1 and Me3 genes, for their resistance to a panel of M. arenaria, M. javanica and M. incognita isolates. The M. arenaria and M. javanica isolates showed low pathogenicity to pepper, unlike the M. incognita isolates. The QR, controlled by the pepper genetic background, displayed a high resistance level with a broad spectrum of action, protecting pepper against Me3‐virulent as well as avirulent M. incognita isolates. The QR was also expressed when combined with the Me1 and Me3 genes, but presented additive genetic effects so that heterozygous F1 hybrids proved less resistant than homozygous inbred lines. The discovery of this QR is expected to provide promising applications for preserving the efficiency and durability of nematode resistance.  相似文献   

10.
Root-knot nematodes (Meloidogyne spp.) are a major problem facing crop production globally including potatoes. During the 2011/2012 potato growing season, root-knot nematode infected potato tubers were obtained from different potato growing regions in South Africa for identification of Meloidogyne spp. Using the intergenic region of the ribosomal DNA (IGS-rDNA) together with the region between the cytochrome oxidase small subunit II (COII) and the 16S rRNA gene in the mitochondrial DNA (mtDNA), five of the 78 composite samples received produced amplicon sizes of 705 bp for COII and 780 bp for IGS typical of M. enterolobii. These five samples were from the KwaZulu-Natal potato producing region. Nucleotide sequencing and phylogenetic analysis of the COII and IGS fragment showed that the five Meloidogyne populations were 100 % similar and they clustered closely with those of M. enterolobii in the GenBank database. The high damage potential of resistance-breaking populations of Meloidogyne species is a threat to profitable potato production and will require effective pest management programmes to be put in place.  相似文献   

11.
Root-knot nematodes (Meloidogyne spp.) are important pests of numerous crops worldwide. Some members of this genus have a quarantine status, and accurate species identification is required to prevent further spreading. DNA barcoding is a method for organism identification in non-complex DNA backgrounds based on informative motifs in short DNA stretches (≈600 bp). As part of the EU 7th Framework project QBOL, 15 Meloidogyne species were chosen to compare the resolutions offered by two typical DNA barcoding loci, COI and COII, with the distinguishing signals produced by two ribosomal DNA genes (small and large subunit rDNA; SSU?≈?1,700 and LSU?≈?3,400 bp). None of the four markers distinguished between the tropical species Meloidogyne incognita, M. javanica and M. arenaria. Taking P ID (Liberal) values ≥0.93 as a measure for species delimitation, the four mtDNA and rDNA markers performed well for the tropical Meloidogyne species complex, M. enterolobii, M. hapla, and M. maritima. Within cluster III A (Holterman et al. Phytopathology, 99, 227–235, 2009), SSU rDNA did not offer resolution at species level. Both mtDNA loci COI and COII did, whereas for LSU rDNA a longer fragment (≥700 bp) is required. The high level of mitochondrial heteroplasmy recently reported for M. chitwoodi (Humphreys-Pereira and Elling Nematology, 15, 315–327, 2013) was not found in the populations under investigation, suggesting this could be a regional phenomenon. For identification of RKNs, we suggest the combined use of SSU rDNA with one of three other markers presented here.  相似文献   

12.
Nine populations of Meloidogyne spp. from Greece have been identified as M. javanica or M. incognita using either isozyme phenotypes or the sequence characterized amplified region-polymerase chain reaction (SCAR-PCR) technique. Virulence against the Mi resistance gene was assayed by pot experiments in controlled conditions and revealed the ability of five populations of M. javanica and one population of M. incognita to reproduce on tomato cultivars containing that gene. A resistance-breaking population of M. incognita is reported for the first time in the country; the M. javanica populations constitute new records for the Greek mainland.  相似文献   

13.
A molecular‐based assay was employed to analyse and accurately identify various root‐knot nematodes (Meloidogyne spp.) parasitizing potatoes (Solanum tuberosum) in South Africa. Using the intergenic region (IGS) and the 28S D2–D3 expansion segments within the ribosomal DNA (rDNA), together with the region between the cytochrome oxidase subunit II (COII) and the 16S rRNA gene of the mtDNA, 78 composite potato tubers collected from seven major potato growing provinces were analysed and all Meloidogyne species present were identified. During this study, M. incognita, M. arenaria, M. javanica, M. hapla, M. chitwoodi and M. enterolobii were identified. The three tropical species M. javanica, M. incognita and M. arenaria were identified as the most prevalent species, occurring in almost every region sampled. Meloidogyne hapla and M. enterolobii occurred in Mpumalanga and KwaZulu‐Natal, respectively, while M. chitwoodi was isolated from two growers located within the Free State. Results presented here form part of the first comprehensive surveillance study of root‐knot nematodes to be carried out on potatoes in South Africa using a molecular‐based approach. The three genes were able to distinguish various Meloidogyne populations from one another, providing a reliable and robust method for future use in diagnostics within the potato industry for these phytoparasites.  相似文献   

14.
Commercial vineyards in southern Spain were surveyed and sampled during October to December 2004 to determine the extent to which common weeds present were suitable hosts of root-knot nematodes infesting soils of those vineyards. Seven weed species commonly growing in grapevine soils in southern Spain were found infected by either Meloidogyne incognita or M. javanica: Amaranthus retroflexus (redroot pigweed), Anchusa azurea (ox-tongue), Chenopodium album (goosefoot), Erodium moschatum (musk stork’s bill), Malva rotundifolia (low mallow), Sinapis alba (white mustard), and Solanum nigrum (black nightshade). The host suitability of the weeds to root-knot nematodes was evaluated on the basis of root galling severity and nematode population densities in soil and roots. Also, the host–parasite relationship in these naturally Meloidogyne-infected weeds was examined. All the weed species in the study were considered suitable hosts for M. incognita and M. javanica because: (a) high Meloidogyne spp. populations occurred in roots and surrounding soil of the weed species; (b) the severity of root galling was high, and (c) well-established permanent feeding sites were observed in the histopathological studies of infected root tissues. In addition, this study presents the first reports of S. alba and A. azurea as hosts for M. incognita, and of E. moschatum as a new host for M. javanica, thus increasing the list of reported weed hosts for Meloidogyne spp. These results indicate that noticeable population densities of M. incognita and M. javanica can be maintained or increased in these weeds, at population levels higher than those previously reported for the same nematodes infecting grapevine roots. The weeds infesting vineyards thus represent an important source of inoculum of Meloidogyne spp., and furthermore may act as reservoirs of these nematodes which can be disseminated within or among vineyards by agricultural operations.  相似文献   

15.
Meloidogyne ethiopica is an important nematode pathogen causing serious economic damage to grapevine in Chile. In Brazil, M. ethiopica has been detected with low frequency in kiwifruit and other crops. The objectives of this study were to evaluate the intraspecific genetic variability of M. ethiopica isolates from Brazil and Chile using AFLP and RAPD markers and to develop a species‐specific SCAR‐PCR assay for its diagnosis. Fourteen isolates were obtained from different geographic regions or host plants. Three isolates of an undescribed Meloidogyne species and one isolate of M. ethiopica from Kenya were included in the analysis. The results showed a low level of diversity among the M. ethiopica isolates, regardless of their geographical distribution or host plant origin. The three isolates of Meloidogyne sp. showed a high homogeneity and clustered separately from M. ethiopica (100% bootstrap). RAPD screenings of M. ethiopica allowed the identification of a differential DNA fragment that was converted into a SCAR marker. Using genomic DNA from pooled nematodes as a template, PCR amplification with primers designed from this species‐specific SCAR produced a fragment of 350 bp in all 14 isolates of M. ethiopica tested, in contrast with other species tested. This primer pair also allowed successful amplification of DNA from single nematodes, either juveniles or females and when used in multiplex PCR reactions containing mixtures of other root‐knot nematode species, thus showing the sensitivity of the assay. Therefore, the method developed here has potential for application in routine diagnostic procedures.  相似文献   

16.
Plant-parasitic nematodes are serious pests causing important crop losses worldwide. After extensive screening of non-tuber-bearing Solanaceae, a resistant trap crop, Solanum sisymbriifolium, with a high production level of hatching agents, seemed an ideal control method for potato cyst nematodes (PCN), Globodera spp. Recently, root-knot nematodes (RKN), Meloidogyne spp., were found coexisting with PCN. Therefore, it is important to find alternative methods to control both nematode genera. The chemical properties of S. sisymbriifolium turns this plant into an excellent candidate for further nematicidal studies and to develop new crop production models. Studies concerning the effects of this plant on plant-parasitic nematodes are presented. Pathogenicity studies with four S. sisymbriifolium cvs (Domino, Pion, Sis 4004 and Sharp) and five Meloidogyne species showed that all cultivars of S. sisymbriifolium studied were resistant to M. chitwoodi and hypersusceptible to M. arenaria and M. hapla. For M. hispanica only cv Pion was susceptible. M. javanica induced different responses: cvs Pion and Sharp were susceptible; cv Domino resistant and Sis 4004 hypersusceptible. The studies of the hatching effects of root exudates from these cvs showed that they had an influence on the hatching inhibition of second stage juveniles of the five Meloidogyne species tested.  相似文献   

17.
The life cycle of a Portuguese Meloidogyne hispanica isolate on susceptible cv. Easypeel and resistant (Mi‐1.2 gene) cv. Rossol tomato plants was studied in growth chambers at constant temperatures (10–35°C). The development within the egg and hatching were compared to those of a Portuguese M. arenaria isolate. The base temperature was 10·11 and 8·31°C with 179·5 and 235·3 thermal units for M. hispanica and M. arenaria, respectively, suggesting better potential adaptation to low temperatures by M. arenaria than M. hispanica. No egg development occurred at 10 or 35°C. An increase in invasion of tomato roots by M. hispanica second‐stage juveniles (J2s) was correlated with an increase in temperature on both tomato cultivars. Tomato cv. Rossol limited M. hispanica development at 20, 25 and 30°C, but not at 35°C, indicating that these high temperatures blocked the resistance mechanism provided by the Mi‐1.2 gene. At 15°C, J2s penetrated tomato cv. Rossol roots, but failed to develop and establish feeding sites. On tomato cv. Easypeel, nematode development and reproduction occurred at 20, 25 and 30°C, but at 20°C the life cycle was 1·5 and 2·0 times longer than at 25 and 30°C, respectively. No egg production was observed at 15°C. The results of this study showed that M. hispanica is most suited to soil temperatures around 25°C. Predicted climate change might favour the spread of this nematode species into southern Europe and northwards. The thermal requirements for M. hispanica development are analysed and compared with those of M. arenaria, M. hapla, M. incognita and M. javanica.  相似文献   

18.
Pot experiments were carried out to characterize the response of two Cucumis metuliferus accessions (BGV11135 and BGV10762) against Mi1.2 gene (a)virulent Meloidogyne arenaria, M. incognita and M. javanica isolates and to determine the compatibility and the effect on physicochemical properties of fruit melons. In addition, histopathological studies were conducted. One week after transplanting, plants were inoculated with one J2 cm?3 of sterilized sand (200 cm3 pots) and maintained in a growth chamber at 25 °C for 40 days. The susceptible cucumber cv. Dasher II or melon cv. Paloma were included for comparison. The number of egg masses and number of eggs per plant were assessed, and the reproduction index (RI) was calculated as the percentage of eggs produced on the C. metuliferus accessions compared to those produced on the susceptible cultivars. The compatibility and fruit quality were assessed by grafting three scions, two of Charentais type and one of type piel de sapo, under commercial greenhouse conditions. The resistance level of both C. metuliferus accessions ranged from highly resistant (RI < 1%) to resistant (1% ≤ RI ≤ 10%) irrespective of Meloidogyne isolates. Melon plants grafted onto C. metuliferus accession BGV11135 grew as self‐grafted plants without negatively impacting fruit quality traits. Giant cells induced by Meloidogyne spp. on C. metuliferus were in general poorly developed compared to those on cucumber. Furthermore, necrotic areas surrounding the nematode were observed. Cucumis metuliferus accession BGV11135 could be a promising melon rootstock to manage Meloidogyne spp., irrespective of their Mi1.2 (a)virulence, without melon fruit quality reduction.  相似文献   

19.
土壤中南方根结线虫的实时荧光PCR检测和定量   总被引：3，自引：2，他引：1  
为了准确测定土壤中南方根结线虫Meloidogyne incognita的群体密度,根据序列特异性扩增区标记设计了南方根结线虫特异性引物和TaqMan探针,分别建立了卵和2龄幼虫的实时荧光PCR定量检测体系,并将该检测体系与显微镜计数法进行了比较.结果表明,引物Mi-F/R及探针Mi-probe对南方根结线虫具有高度特异性,建立的标准曲线循环阈值与卵和2龄幼虫数量的对数值之间具有良好的线性关系,相关系数分别为0.9771和0.9853,扩增效率均为90％,检测灵敏度为10-1个卵及10-3条2龄幼虫;对10个田间土壤样本的检测显示,实时荧光PCR定量检测与显微镜计数法测定的南方根结线虫卵和2龄幼虫数量呈显著正相关,且两种方法对2龄幼虫的测定结果无显著差异.  相似文献   

20.
The response of four Mi-resistance gene tomato rootstocks to seven populations of Meloidogyne was determined in pot tests conducted in a glasshouse. Rootstocks PG76 (Solanum lycopersicum × Solanum sp.) and Brigeor (S. lycopersicum × S. habrochaites) and resistant cv. Monika (S. lycopersicum) were assessed against one population of M. arenaria, three of M. incognita, and three of M. javanica. Rootstocks Beaufort and Maxifort were assessed against one population of M. arenaria, two of M. incognita and two of M. javanica. Rootstock PG76 was highly resistant (reproduction index <10%) to all the populations, whereas rootstock Brigeor and cv. Monika were highly to moderate resistant. Rootstocks Beaufort and Maxifort showed reduced resistance or inability to suppress nematode reproduction, and their responses varied according to the population tested. Beaufort and Maxifort were susceptible to the two populations of M. javanica as Maxifort was to one of M. incognita. The reproduction index of the nematode was higher (P < 0.05) on Maxifort than Beaufort for all root-knot nematode populations.  相似文献   

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