Resistance of Brassicaceae plants to root-knot nematode (Meloidogyne spp.) in northern Australia

Abstract

Brassicaceae plants have the potential as part of an integrated approach to replace fumigant nematicides, providing the biofumigation response following their incorporation is not offset by reproduction of plant-parasitic nematodes on their roots. Forty-three Brassicaceae cultivars were screened in a pot trial for their ability to reduce reproduction of three root-knot nematode isolates from north Queensland, Australia: M. arenaria (NQ1), M. javanica (NQ2) and M. arenaria race 2 (NQ5/7). No cultivar was found to consistently reduce nematode reproduction relative to forage sorghum, the current industry standard, although a commercial fodder radish (Raphanus sativus) and a white mustard (Sinapis alba) line were consistently as resistant to the formation of galls as forage sorghum. A second pot trial screened five commercially available Brassicaceae cultivars, selected for their biofumigation potential, for resistance to two nematode species, M. javanica (NQ2) and M. arenaria (NQ5/7). The fodder radish cv. Weedcheck, was found to be as resistant as forage sorghum to nematode reproduction. A multivariate cluster analysis using the resistance measurements, gall index, nematode number per g of root and multiplication for two nematode species (NQ2 and NQ5/7) confirmed the similarity in resistance between the radish cultivar and forage sorghum. A field trial confirmed the resistance of the fodder radish cv. Weedcheck, with a similar reduction in the number of Meloidogyne spp. juveniles recovered from the roots 8 weeks after planting. The use of fodder radish cultivars as biofumigation crops to manage root-knot nematodes in tropical vegetable production systems deserves further investigation.     

Plant parasitic nematodes associated with olive1

Over 70 species, belonging to 33 genera, of plant parasitic nematodes have been reported in association with olive. Several species of Helicotylenchus have been observed to cause root necrosis and considered capable of affecting olive tree growth. Three species of Meloidogyne (M. incognita, M. lusitanica and M. javanica) were responsible for heavy galling and growth retardation in inoculation trials and natural infestation. Pratylenchus vulnus has been demonstrated in inoculation trials as a potential pathogen of olive. Xiphinema species commonly occur around olive roots and X. elongatum has been shown to affect olive plant growth. Several sedentary plant nematodes attack olive. Tylenchulus semipenetrans infects olive in California (US) and Italy. Trophotylenchulus saltensis was described from olive roots in Jordan and a cyst nematode, Heterodera mediterranea, was shown to feed and multiply on olive roots. The cortical feeding cells of roots fed upon by Gracilacus peratica showed thickened and lignified wall near the penetration points of the stylet and those exposed to Ogma rhombosquamatum also presented hypertrophic nuclei and nucleoli. Rotylenchulus macrodoratus induces the formation of an enlarged mononucleate stelar or cortical ‘nurse cell’ with dense cytoplasm and hypertrophic nucleolus.     

Does soil warming affect the interaction between Pasteuria penetrans and Meloidogyne javanica in tomato plants?

A system to grow tomato plants infected by Meloidogyne javanica under constant temperatures of 18, 21, 24, 27 and 30 °C was developed and used to assess how temperature and the application of the biological control bacterium Pasteuria penetrans affected plant growth, the nematode population and endospore production. Each plant was inoculated with 300 second‐stage juveniles (J2) with four or five spores of P. penetrans attached to their cuticles or with 300 nematodes without P. penetrans. Increasing soil temperature increased tomato growth, the number of endospores per female, and the number of galls of M. javanica at the end of 38 days. Increasing temperatures up to 27 °C also increased the number of egg masses produced by M. javanica. Presence of P. penetrans reduced the numbers of galls and egg masses at all temperatures by up to 52.2% and 61.4% at 27 and 30 °C, respectively. Pasteuria penetrans reduced the M. javanica population even at soil temperatures of 18 and 21 °C. However, temperatures of 27 and 30 °C enhanced nematode control and the production of P. penetrans endospores is faster. The system developed in this work is simple and efficient for growing plants under constant temperatures and can be used for different purposes.     

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

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

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

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

Weed hosts of Meloidogyne spp. and the effect of aqueous weed extracts on egg hatching

Weeds have a great economic impact on agricultural production because they compete with crops for resources and are alternative hosts for pests, microbial pathogens and plant-parasitic nematodes. This study aimed to investigate the susceptibility of weeds to the root-knot nematodes Meloidogyne javanica and Meloidogyne incognita and assess the effect of aqueous weed extracts on M. javanica egg hatching. Four experiments were conducted, two for each nematode species. Weeds were inoculated with 2000 nematode eggs and grown for 60 days under greenhouse conditions. Soyabean cv. Monsoy 7110 was used as control. The weeds Ipomoea grandifolia, Solanum americanum, Digitaria horizontalis, Amaranthus deflexus, Sorghum halepense and Commelina benghalensis were susceptible to M. javanica and M. incognita in at least one experiment (reproduction factor, RF >1). Crotalaria juncea and Eleusine indica were susceptible to M. incognita in one experiment, and Digitaria insularis, Sida rhombifolia, Bidens pilosa, Urochloa decumbens, Crotalaria breviflora, Cenchrus echinatus, Crotalaria ochroleuca and Crotalaria spectabilis were immune or resistant (RF <1 or RF = 0). Alternanthera tenella, C. juncea, S. rhombifolia, C. ochroleuca, C. spectabilis, C. breviflora, B. pilosa, E. indica, U. decumbens and C. echinatus were resistant or immune to M. javanica (RF <1 or RF = 0). Compared with the control (water), all weed extracts reduced M. javanica egg hatch. Our results highlight the importance of weed control in agricultural systems, as nematodes can survive and multiply in weed roots during the off-season. Weed leaves and shoots, however, may be an interesting source of compounds with nematicidal activity.     

Pathogenicity of Meloidogyne incognita and M. javanica on recombinant inbred lines from a crossing of Cucurbita pepo subsp. pepo × C. pepo subsp. ovifera

The response of recombinant inbred lines (RILs) from a cross of zucchini × scallop (Cucurbita pepo subsp. pepo ‘Murcia MU-CU-16’ × C. pepo subsp. ovifera ‘Scallop UPV-196’) to Meloidogyne incognita and M. javanica was determined after completion of a nematode reproduction cycle in experiments carried out in a growth chamber. The nematode differentiated the C. pepo genotypes at the subspecies level due to lower egg mass production on subspecies pepo than ovifera, and thus subspecies pepo was a poorer host than ovifera. In addition, Murcia MU-CU-16 discriminated M. incognita from M. javanica in terms of egg masses (EM), eggs per gram of root and reproduction factor (Rf), whereas Scallop UPV-196 did so in eggs per gram of root and Rf. The RILs differed in gall formation and EM production depending on the nematode × line combination. Comparisons between nematode isolates resulted in four significant combinations for pathogenic potential (galls/initial population (Pi) × 100), seven for parasitic success (egg masses/Pi × 100), and nine for host efficiency (egg masses/galls per root system × 100) which included all the lines tested against both isolates. Lines that restricted nematode development by at least 90% were considered as having intermediate resistance to M. incognita based on the definition of the International Seed Federation. They included lines 28-1, 35A, 107A, 110-3 and 153-2. All the RILs were susceptible hosts for M. javanica. The information presented here will be helpful for nematode management and also for plant breeders working on pathogen resistance on C. pepo.     

Nematicidal Activity of Chrysanthemum coronarium

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

Use of green leaves from olive trees as soil amendment for the control of Meloidogyne1

Plant extracts have been used against nematodes mainly in the third-world countries. The possibility of using leaves of olive trees for reducing populations of nematodes (Meloidogyne spp.) in the soil was investigated in the present work. The root-knot incidence in tomato roots was reduced as well as the presence of nematodes in the soil. The methanol extract of the leaves inhibited hatching of the eggs almost completely.     

Carbon Fixation and Partitioning in Coffee Seedlings Infested with <Emphasis Type="Italic">Pratylenchuscoffeae</Emphasis>

The objective was to study CO₂ fixation and photoassimilate partition in coffee (Coffea arabica) seedlings infested with the lesion nematode Pratylenchus coffeae. Seedlings infested with 0, 1000 and 8000 Pratylenchus coffeae nematodes were exposed to ¹⁴CO₂ and the incorporation and distribution of radioactivity were followed in the roots, stems and leaves. Fresh mass, pigments, soluble sugars, sucrose and specific radioactivity of sucrose in the plant parts were determined. At the highest level of infestation almost all the parameters were significantly changed showing the carbon fixation in the leaves and partitioning to the roots were decreased. Since lesion nematodes are not sedentary and do not form feeding sites that could be characterised as metabolic sinks, it is suggested that their damage is more readily expressed by the leaves, through a reduction in photosynthesis and phloem transport.     

Susceptibility of hairy root lines of Brassica species to Plasmodiophora brassicae and in an in vitro subculture system

To investigate the susceptibility of hairy root lines of Brassica species to Plasmodiophora brassicae, hairy roots were induced in a number of Brassica species with Agrobacterium rhizogenes. Turnip hairy root was highly susceptible to P. brassicae; infection rates were high and large galls formed. In contrast, the rates of root hair infection and gall formation on intact Brassica plants did not differ significantly from the control. To induce resting spore formation, turnip hairy roots were incubated at 15°, 20°, or 25°C after 3 weeks of incubation at 25°C. The number and fresh mass of the galls per hairy root were higher and formation of resting spores was greatest after a 7-week incubation at 20°C. To subculture P. brassicae using turnip hairy root, turnip hairy roots were reinoculated with resting spores and gall with resting spores then formed on the hairy roots. In this way, P. brassicae using hairy roots could be subcultured in vitro two or three times on three single-spore isolates of P. brassicae. This is the first report of in vitro subculture of P. brassicae using hairy root.     

Molecular cloning and virus-induced gene silencing of MiASB in the southern root-knot nematode, Meloidogyne incognita

Root-knot nematodes (Meloidogyne spp.), cause serious damage to agricultural production worldwide. In this study, we designed special primers based on the predicted Mitochondrial ATP synthase b subunit gene (ASB) sequence to clone the same gene in M. incognita (MiASB). The identity between the cloned MiASB and the predicted MiASB was as high as 100 %. Using the tobacco rattle virus (TRV)-mediated virus-induced gene silencing (VIGS) system, we delivered MiASB RNAi triggers to the M. incognita feeding site on tomato seedlings, resulting in significantly fewer galls on the seedlings. Sixty days after inoculation with M. incognita, the number of root galls induced on the MiASB silence-treated seedlings was reduced by 64.3 % compared to that on the control seedlings, and reduced by 64.1 % compared to that on untreated control seedlings. This study revealed the MiASB silencing had a positive effect on the control of root-knot nematodes, and MiASB may be associated with the formation of galls caused by the nematode.     

Crown gall of tobacco caused by <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> biovar 1 in tobacco fields

Crown gall disease of tobacco was found in Iwate Prefecture, Japan in 1995. Ten bacterial isolates, obtained from the galls of tobacco, were identified as Agrobacterium tumefaciens (Smith and Townsend 1907) Conn 1942 biovar 1 based on their ability to induce galls on the 14 tested plants, including tobacco after needle-prick inoculation, and on 12 cultural, physiological, and biological characteristics. The growth of the causal organism was not inhibited in vitro by agrocin of A. radiobacter strain K84. This report is the first on the natural occurrence of crown gall caused by A. tumefaciens on tobacco plants.     

Host suitability of Vitis rootstocks to root‐knot nematodes (Meloidogyne spp.) and the dagger nematode Xiphinema index,and plant damage caused by infections

The host suitability of commercial Vitis rootstocks commonly used in Spain (161‐49C, 41B, 1103P, 110R, 140Ru and SO4) to root‐knot nematodes (Meloidogyne arenaria, M. incognita, M. javanica) and Xiphinema index, and damage caused by nematode infection were determined under controlled conditions. The three root‐knot nematodes reproduced with a rate higher than one in all rootstocks, indicating that they are suitable hosts for these nematodes. Growth of rootstocks infected with the root‐knot nematodes was less vigorous than that of nematode‐uninfected controls in the majority of the rootstocks studied. Root infection resulted in moderate to severe root galling in all rootstocks. The shoot and main stem diameters appeared to be the most sensitive variables of damage caused by infection by Meloidogyne spp., with reduction rates from 36% and 53% in 161‐49C to 57% and 66% in 140Ru, respectively. The shoot height was not significantly affected by the root‐knot nematodes and the root fresh weight generally increased as a consequence of intensive galling. The nematode X. index caused significant root damage with a reproduction factor higher than one in all rootstocks. However, reproduction factor was significantly influenced by the rootstock and significantly decreased by about 12‐fold (5·7 to 18·1‐fold) with the increase in inoculum density from 100 to 1000 nematodes per plant. The root dry weight was reduced by X. index infections, and was the plant growth variable most affected by the nematode infection in all rootstocks at both inoculum densities. Meloidogyne arenaria, M. incognita, M. javanica and X. index, prevalent in many world vineyards, are all shown to have a damaging effect on the six tested rootstocks.     

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

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

Occurrence of virulent root-knot nematode populations on tomatoes bearing the <Emphasis Type="Italic">Mi</Emphasis> gene in protected vegetable-growing areas of Turkey

Root-knot nematodes (Meloidogyne spp.; RKN) are one of the most important pathogens of vegetables in Turkey. Assessing the existing virulent RKN populations is of importance for pathogen mapping in the west Mediterranean region of Turkey. Therefore, 95 populations of RKN were collected from different protected vegetable-growing locations in the region. Pure cultures were obtained and identified by means of species-specific primers. Virulence of the populations against the Mi-1 gene conferring resistance to Meloidogyne incognita, M. javanica and M. arenaria was determined according to their egg masses and gall rating on resistant and susceptible tomato varieties. Results showed that seven populations of M. incognita and six populations of M. javanica were able to overcome the resistance controlled by the Mi-1 gene. The frequency of virulent populations of M. incognita and M. javanica collected from different protected-grown vegetables was 11.7% and 21.4%, respectively. To our knowledge, this is the first report of populations of RKN virulent to the Mi-1 gene in Turkey.