Plant resistance to aphid feeding: behavioral,physiological, genetic and molecular cues regulate aphid host selection and feeding

Aphids damage major world food and fiber crops through direct feeding and transmission of plant viruses. Fortunately, the development of many aphid‐resistant crop plants has provided both ecological and economic benefits to food production. Plant characters governing aphid host selection often dictate eventual plant resistance or susceptibility to aphid herbivory, and these phenotypic characters have been successfully used to map aphid resistance genes. Aphid resistance is often inherited as a dominant trait, but is also polygenic and inherited as recessive or incompletely dominant traits. Most aphid‐resistant cultivars exhibit constitutively expressed defenses, but some cultivars exhibit dramatic aphid‐induced responses, resulting in the overexpression of large ensembles of putative aphid resistance genes. Two aphid resistance genes have been cloned. Mi‐1.2, an NBS‐LRR gene from wild tomato, confers resistance to potato aphid and three Meloidogyne root‐knot nematode species, and Vat, an NBS‐LRR gene from melon, controls resistance to the cotton/melon aphid and to some viruses. Virulence to aphid resistance genes of plants occurs in 17 aphid species – more than half of all arthropod biotypes demonstrating virulence. The continual appearance of aphid virulence underscores the need to identify new sources of resistance of diverse sequence and function in order to delay or prevent biotype development. © 2013 Society of Chemical Industry     

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.     

Dry mycelium of<Emphasis Type="Italic">Penicillium chrysogenum</Emphasis> protects cucumber and tomato plants against the root-knot nematode<Emphasis Type="Italic">Meloidogyne javanica</Emphasis>

Incorporation into soil of dry mycelium ofPenicillium chrysogenum, a waste product of the pharmacological industry, enhanced plant growth and reduced root galling caused by the root-knot nematodeMeloidogyne javanica in cucumber and tomato plants. Incorporation into sandy loam soil in pots of dry mycelium at a concentration of 0.25% (w/w) resulted in complete protection of cucumber plants from the nematode. The number of juveniles recovered from soils containing dry mycelium was greatly reduced even at a concentration of 0.1% (w/w). In microplot studies conducted at two sites in two seasons, with three or four doses, dry mycelium caused a dose-dependent reduction in root galling index (GI) and promotion of plant growth of cucumber and tomato plants. Inin vitro studies, the water extract of dry mycelium immobilized nematode juveniles and reduced the egg hatching rate, but these effects were partly reversible after a rinse in water. Soil-drenching of cucumber and tomato seedlings with water extract of dry mycelium did not reduce GI or number of root-invading juveniles. The results show that dry mycelium promotes plant growth and protects plants against nematode infection. Protection, however, does not operatevia induced resistance. http://www.phytoparasitica.org posting April 6, 2003.     

Expression of the Arabidopsis MCM Gene PROLIFERA During Root-Knot and Cyst Nematode Infection

ABSTRACT Expression of the Arabidopsis thaliana gene PROLIFERA (PRL) was examined during development of root-knot and cyst nematode feeding sites. These obligate plant parasites establish specialized feeding structures in roots that allow them to withdraw nutrients from the host. In the process of establishing feeding sites, nematodes alter cell cycle regulation. PRL is normally expressed specifically in dividing cells at all stages of plant development and was used here as a marker for cell division. PRL expression, reported from a PRL::GUS fusion protein, was detected in nematode feeding sites of both root-knot and cyst nematodes from the earliest stages of infection in both giant cells and syncytia. However, unlike other cell cycle genes, expression of PRL was detected only occasionally in cells surrounding the feeding sites. PRL::GUS activity persisted until late in the infection cycle, past the time when other cell cycle genes are expressed. These data indicate that some aspects of the PRL expression pattern during nematode infection differ from that of other cell cycle genes.     

Adverse effect of the chitinolytic enzyme PjCHI‐1 in transgenic tomato on egg mass production and embryonic development of Meloidogyne incognita

A novel chitinase gene (PjCHI‐1) isolated from Paecilomyces javanicus, a non‐nematophagous fungus, and driven by a CaMV35S promoter, was delivered into CLN2468D, a heat‐tolerant cultivar of tomato (Solanum lycopersicum). T₁ tomato plants exhibited high endochitinase activity and reduced numbers of eggs and egg masses when infected with the root‐knot nematode (RKN) Meloidogyne incognita. The eggs found in transgenic tomato had lower shell chitin contents than eggs collected from control plants. Egg masses from transgenic plants exhibited higher chitinase activity than those from control plants. Moreover, only 30% of eggs from transgenic plants were able to develop to the multi‐cell/J1 stage, compared with more than 96% from control plants. The present study demonstrated that the expression of the PjCHI‐1 chitinase gene can effectively reduce the production of egg masses and repress the embryonic development of M. incognita, presenting the possibility of a novel agro‐biotechnological strategy for preventing crop damage by RKN.     

Biocontrol traits of plant growth suppressive arbuscular mycorrhizal fungi against root rot in tomato caused by <Emphasis Type="Italic">Pythium aphanidermatum</Emphasis>

Arbuscular mycorrhizal (AM) fungi known to cause plant growth depressions in tomato were examined for their biocontrol effects against root rot caused by Pythium aphanidermatum. The main hypothesis was that plant growth suppressive AM fungi would elicit a defence response in the host plant reducing Pythium root rot development. To test this hypothesis a fully factorial experiment was performed with AM fungi (Glomus intraradices, G. mosseae, G. claroideum or nonmycorrhizal), Pythium (± P. aphanidermatum) and harvest (7 and 14 days after pathogen inoculation (dapi)) as the main factors. Two weeks after AM fungi inoculation, roots were challenged with P. aphanidermatum. Variables evaluated at each harvest were root colonization levels of the interacting fungi, plant growth responses, and expression of a plant pathogenesis related protein gene (PR-1). All of the tested AM fungi caused marked growth suppressions, but did not affect PR-1 gene expression or the phosphorous concentration in the host plant. Plants singly inoculated with P. aphanidermatum had an increased PR-1 expression and phosphorous concentration. Among the AM fungi included in the study only G. intraradices reduced the pathogen root infection level, measured both in terms of Pythium ELISA and by recovery on selective media and only at the first harvest. Likewise, P. aphanidermatum root infection reduced colonization levels of G. intraradices, but not that of the two other AM fungi. In conclusion, plant growth suppressive AM fungi may offer plant beneficial traits in terms of biocontrol of root cortical pathogens.     

Biological and molecular characterization of the response of tomato plants treated with Trichoderma koningiopsis

Tomato-Fusarium oxysporum f.sp. radicis-lycopersici pathosystem was used to study induced systemic resistance elicited by Trichoderma koningiopsis (Th003) using the split root model. The ability of the antagonist to promote plant growth was also established. Stem colonization by the pathogen was significantly reduced in treated plants. The induction of resistance was enhanced 6 days after elicitation and when the antagonist was used in a concentration of 10⁵ conidia per ml. Th003 application in seed priming and nursery significantly stimulated plant growth. Gene expression induced by Th003 was evaluated using the tomato TOM1 microarray. Plant treatment with T. koningiopsis affected mRNA levels of 45 genes: 41 in roots and 4 in leaves. Of particular interest was the induction of genes involved in the jasmonic and ethylene transduction pathways found in the microarray analysis and qRT-PCR, which suggest a temporary increment of defense related gene expression response to T. koningiopsis Th003.     

膏体迷向剂对苹果园梨小、桃小食心虫的防效

2013年在宁夏苹果园进行了复合膏体迷向剂对梨小、桃小食心虫防效的进一步试验,试验设3个处理(涂抹高度3.5m、3.5m/4.5m交叉涂抹、交叉涂抹空白区域)、1个对照,2种膏剂附着方式(树干附着、膏剂填装塑料瓶悬挂)。通过监测全年诱蛾量、调查果实膨大期与成熟期蛀果率分析防控效果。结果显示,复合式膏体迷向剂可有效防止梨小食心虫对果实的为害,涂抹高度3.5m时,梨小食心虫诱蛾量下降76.68%;交叉涂抹时,下降70.8%;交叉涂抹空白区域下降36.8%。3个处理成熟期蛀果率分别下降84.88%、91.39%、24.19%。由于试验区桃小食心虫种群密度小,试验无法确定该迷向剂对桃小食心虫的防治效果。     

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.     

The Exploitation of Nematode-Responsive Plant Genes in Novel Nematode Control Methods

The molecular interactions between plants and sedentary nematodes are undergoing intense study, not only for reasons of fundamental research but also for the potential benefits to agriculture. The present technology allows the transformation of an increasing number of crop plants, providing new ways to introduce resistance against plant-parasitic nematodes. The ability of sedentary nematodes to induce specialized feeding sites in plant roots is one of the most fascinating aspects of this host–parasite interaction. Molecular approaches have been initiated to identify and characterize plant genes altered in expression after infection by sedentary nematodes. The results obtained indicate that many genes indeed become up-regulated upon nematode infection. Surprisingly, several so-called constitutive promoters that are normally used to achieve high expression in plant cells are completely ‘silenced’ in the feeding sites within days after nematode infection. Generally, there are two options available for the genetic engineering of nematode resistance: the synthesis of anti-nematode proteins or the localized production of a cytotoxic protein that interferes with the development of feeding cells. Nematode-induced promoters are very useful for the production by plants of sufficiently high levels of anti-nematode proteins at feeding sites. Alternatively, interfering with feeding-cell development is somewhat similar to the hypersensitive response evoked by nematodes in a naturally resistant plant. Here, destruction of specific plant cells can be achieved by the localized expression of a cytotoxin such as barnase, a potent ribonuclease. This approach, however, calls for a highly specific ‘non-leaky’ promoter, which is active only in the feeding cells. Another possibility is to use a two-component system, where the leakiness of the promoter in other tissues is counterbalanced by the constitutive expression of a neutralizing gene.     

Nematicidal activity of the leaf powder and extracts of Myrtus communis against the root‐knot nematode Meloidogyne javanica

Nematicidal activity of the leaf powder, leaf extracts and formulated leaf extracts of Myrtus communis, an evergreen shrub that is widely distributed in Israel and other Mediterranean countries, was evaluated using the root‐knot nematode Meloidogyne javanica in in vitro and pot experiments. Leaf powder added to sand at 0·1% (w/w) reduced the number of juveniles recovered from the sand by more than 50%. Reduction in galling index and number of nematode eggs on tomato roots was also observed by incorporating the leaf powder at 0·1–0·4% (w/w) in the soil in pot experiments. Leaf powder extracts with methanol or ethanol showed the highest nematicidal activity among all extracts tested. Emulsifiable concentrates of leaf‐paste extract at a concentration as low as 0·005% (a.i., w/w) reduced the number of juveniles recovered from treated sand and the gall index of cucumber seedlings. The extract paste at 26 g m^?2 was also effective in reducing the gall index of tomato plants in field‐plot experiments. The leaf powder at 0·2% and the formulated leaf‐paste extract at 0·02% were also nematicidal to Tylenchulus semipenetrans and Ditylenchus dipsaci, but not to Pratylenchus penetrans or Steinernema feltiae. At least three nematicidal compounds were found in the leaf extract upon fractionation by thin‐layer chromatography. The results suggest that the leaf powder and paste extract of M. communis are potential nematicides against root‐knot nematodes.     

Histopathology of roots of three tomato cultivars infected with two separate isolates of the false root-knot nematode <Emphasis Type="Italic">Nacobbus aberrans</Emphasis>

The plant-parasitic nematode Nacobbus aberrans attacks weeds and cultivated plants, causing drastic crop yield losses. Several tomato cultivars, such as Superman and Mykonos, are produced by Seminis® as nematode resistant and are widely used in Argentina; their specific resistance response to different nematode species, however, is still unknown. In this study we explored the response of tomato cultivars to the infection of N. aberrans isolates, and determined the host status by performing histological analyses and estimating egg mass index (EMI). Two Argentina isolates (from Lules-Tucumán and Río Cuarto-Córdoba) were tested separately in plants of Superman, Mykonos and Platense cultivars. Plants were maintained in a greenhouse for 90 days; then EMI was estimated in root systems and the material was processed to prepare histological slides and for histochemical test. Infected roots exhibited galls with females and a syncytium (feeding site) developed inside them. The vascular tissues were disorganized and displaced to the periphery; xylem percentage was lower than that in the control plants. All the cultivars were susceptible and developed a close plant-parasite relationship, with Mykonos and Platense cultivars infected with the Lules isolate being the most highly affected, as indicated by their highest EMI values. Superman was the least susceptible cultivar, as evidenced by its lowest EMI values, the amount of starch observed, the presence of thickened cell walls around the nematode and the egg mass, and the low percentage of gall occupied by syncytium.     

Post‐infection development and histopathology of Meloidogyne arenaria race 1 on Arachis spp.

The reproductive behaviour of the root‐knot nematode Meloidogyne arenaria race 1 was compared on two wild species of Arachis (A. duranensis and A. stenosperma) and cultivated peanut (A. hypogaea cv. IAC‐Tatu‐ST). The three species were considered moderately susceptible, resistant, and susceptible, respectively. Penetration and development of the root‐knot nematode in the resistant species was reduced in comparison with that occurring in susceptible plants. Several cell features, including dark blue cytoplasm and altered organelle structure were observed in the central cylinder of A. stenosperma, indicating a hypersensitive‐like response (HR) of infested host cells. Neither giant cells, nor nematodes developed beyond the second stage, were found on A. stenosperma. Arachis duranensis showed a delay in the development of nematodes in the roots compared to A. hypogaea. The two wild peanut species were chosen to be the contrasting parents of a segregating population for mapping and further investigation of resistance genes.     

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.