Glandular hairs providing resistance to aphids in certain wild potato species

Glandular hairs occur abundantly on Solanum polyadenium, S. tarijense and S. berthaultii. An exudate is discharged from the glandular hair when an aphid mechanically ruptures the cell walls. On contact with atmospheric oxygen, a chemical reaction is initiated in which the clear, water-soluble material contained in the hair changes to a black insoluble material which is precipitated on the aphid's limbs. This initially impedes its movements and further accumulations eventually stick the aphids to the plant so that they are immobilized and starve to death. In experiments described, about 30 % of infesting aphids were completely immobilized within 24 h. The potential value of this source of resistance to aphids, if incorporated into the cultivated potato, is discussed, especially with reference to the advantages of its being combined with other forms of resistance and to its effectiveness against virus vectors.     

Chemistry of the secretion from the cornicles of various species of aphids

The cornicle secretions from 28 species of aphids were examined directly by mass spectrometry and, in selected cases, by chemical and chromatographic methods. The major components of the secretions are triglycerides, the acid moieties of which are hexanoic, hexadienoic (sorbic), tetradecanoic (myristic), and hexadecanoic (palmitic) acids, with smaller amounts of C₁₂ and C_18:1 acids. The mixtures from the different species fall into six groups which are readily distinguished by mass spectrometry. The composition was independent of the host plant or of the stage of development where these have been investigated. Biochemical classification is not consistent with morphological classification in the species examined.     

Chemical basis of host-plant resistance to aphids

Abstract The importance of host-plant resistance of crop plants in the biological control of aphids and the formation of aphid biotypes is discussed. The rapid response of plant breeders to the formation of new aphid biotypes requires detailed knowledge on the mechanism of host-plant resistance. Current knowledge on the chemical basis of host-plant resistance to aphids is summarized. Of central importance to this relationship is the aphid-pectinase plant-pectin interdependence. Parallels between the mechanism of aphid-plant and pathogen-plant attack are pointed out and how aphid bacterial endo-symbiotes play a central role in host-plant selection.     

Detection of resistance to aphids in cereals

Resistance to Sitobion avenae and Metopolophium dirhodum was assessed by caging aphids on the upper part of mature tillers. Despite variation in the results, considered to be due mainly to differences amongst plants of different ages, certain cultivars consistently supported fewer aphids. Winter wheats Anna Migliori and Marsters Al and Indian spring barleys EB 921 and DL 107 were identified as resistant cultivars useful for reference in further work.     

Alkaloid secretion inhibited by antibiotics in Aphaenogaster ants

Although alkaloids are frequent in the poison glands of ants of the genus Aphaenogaster, this is not the case for A. iberica. Hypothesizing that in the genus Aphaenogaster, alkaloids are produced by symbiotic bacteria, except for A. iberica, we treated an experimental lot of both A. iberica and a ‘classical’ Aphaenogaster species, A. senilis, with an antibiotic. Compared to workers from a control lot, this treatment reduced considerably alkaloid production in A. senilis, whereas A. iberica did not react to the treatment. Furthermore, the treatment induced an increase in cuticular hydrocarbon quantities in A. senilis, but not in A. iberica. An analysis of the ant microbiota will be the next step to confirm our hypothesis.     

Morphology and secretion of glandular trichomes in Tamus communis

Light and scanning electron microscopical investigations were carried out to study distribution, morphology and secretion of glandular trichomes during leaf development in Tamus communis . During leaf growth glandular trichomes arise continously from single protodermal cells. At maturity, they are composed of a 6-celled gland head, a secretory neck cell, an "endodermal" cell, and one basal or reservoir cell. During the early stage of secretion, several regularly arranged crater-like pores differentiate on the cuticular surface of the gland head. Through these pores (1–1.5 μm in diameter), the secretion flows out as a thin film or as rods (0.4 μm in diameter) spreading on the trichomes and on the leaf surface. Histochemical tests indicate that the secretion is composed of a small amount of carbohydrates and an abundant fraction of lipophilic material. Proceeding towards senescence, the wall degeneration of the gland head gives rise to a strong histochemical reaction for insoluble polysaccharides. The results presented are compared with those of other investigations on the pattern of secretion in glandular trichomes.     

Mapping resistance to cereal aphids in barley

 A set of 150 doubled-haploid (DH) barley (Hordeum vulgare L.) lines derived from the cross of Harrington/TR306 was used to determine the number and chromosomal location of quantitative trait loci (QTLs) controlling resistance to cereal aphids. The experiments were conducted under natural infestation in the field during two growing seasons: 1994 and 1995. Aphid resistance was measured by counting the number of aphids per plot. Counts were made on a weekly basis. Each year at the time of maximum aphid density there was an obvious difference in reaction between the parental genotypes. The DH lines showed continuous variation for aphid density. Simple interval mapping and simplified composite interval mapping revealed that the principal QTL determining cereal aphid resistance is on the distal region of the short arm of chromosome 1. This aphid-resistance QTL is linked with a heading-date QTL. At the time of highest aphid infestation, this QTL accounted for 31% and 22% of the total variance of aphid density in 1994 and 1995, respectively. A QTL on chromosome 5 was also detected but only by simplified composite interval mapping. However, the largest consistent effect was due to the QTL on the short arm of chromosome 1. This QTL may be a useful target for marker-assisted selection for adult plant cereal aphid resistance in barley. Received: 10 September 1996/Accepted: 11 October 1996     

Variation of Alkaloid Production in Nicotiana rustica Callus Cultures

Callus cultures have been established from the seed, root and leaf of Nicotiana rustica L. var. brasilia in a synthetic medium containing 1 μM 2,4-D and μM kinetin. These callus tissues behaved similarly not only in growth and organogenesis but also in nicotine production. The nicotine contents of callus cultures, which were in the order of 0.25–0.58% of dry weight during a few passages subsequent to callus induction, rapidly decreased to trace amounts in succeeding subcultures in association with the decline of the root-regenerating activity. On the other hand, free cells prepared from a callus tissue in the third passage developed into individual clones showing wide differences in growth and nicotine production. One of these clones gave rise to a relatively stable strain which is capable of producing nicotine at a high rate (0.29% of dry weight) in the absence of organization. The significance of these findings is discussed in connection with some results which have been reported for other callus cultures of Nicotiana species.     

Biosynthesis of labdenediol and sclareol in cell-free extracts from trichomes of Nicotiana glutinosa

Biosynthesis of the diterpenes labdenediol and sclareol from all trans-geranylgeranyl pyrophosphate was observed in cell-free extracts prepared from leaf midvein epidermal peels of Nicotiana glutinosa 24A. The bioactivities were shown to be exclusively localized in trichomes, to be conferred by a soluble enzyme(s), and to resemble other plant terpene cyclase activities in basic characteristics. Chromatographic methods for protein purification including gel filtration, anion exchange, hydroxyapatite chromatography, and free-flow isoelectric focusing. Thermal inactivation and end-product inhibition experiments did not afford separation of the biosynthetic activities. Results indicate that these labdane diterpenes are direct products of cyclization reactions catalysed by a soluble cyclase(s). To the best of our knowledge, this report represents the first case for direct biosynthesis of diterpene di-alcohols by cyclization of the acyclic precursor. A unified scheme for formation of labdane mono- and di-alcohols in Nicotiana species is presented.Abbreviations DEAE diethylaminoethyl - GGPP geranyl-geranyl pyrophosphate     

Glandular trichomes of Solanum berthaultii and resistance to the Colorado potato beetle

The defensive mechanisms of the wild potato, solanum berthaultii Hawkes, to larvae of the Colorado potato beetle, Leptinotarsa decemlineata (Say), were studied by selective removal of glandular trichomes and trichome exudates from leaflets, and by comparing performance on S. berthaultii and on the cultivated potato, S. tuberosum L., which lacks defensively active type A and B glandular trichomes. Removal of type A trichomes increased the proportion of larvae that fed on S. berthaultii. Removal of the exudate from type B trichomes increased the proportion of larvae that fed and led to a decrease in mortality. The predominant active compounds in type B exudate, i.e. fatty acid esters of sucrose, were only effective in the presence of type A trichomes. Sucrose esters did not affect larval feeding on S. tuberosum leaflets or on S. berthaultii leaf discs from which the type A trichomes had been removed. Growth of surviving larvae was not significantly affected by removing type A trichomes or type B exudate. Growth of larvae was significantly increased when S. berthaultii leaflets were presented in artificial diet which eliminated the physical barrier of the type B stalks. Growth was no different on artificial diet containing either S. berthaultii or S. tuberosum leaf material (fresh or lyophilized powder) but was poorer on these diets than on S. tuberosum leaflets. The presence of type A trichomes is a fundamental requirement for expression of S. berthaultii resistance to L1 L. decemlineata. Type B droplets containing sucrose esters increase the expression of resistance in the presence of defensively-active type A trichomes.

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Résumé Les mécanismes de défense de la pomme de terre sauvage, S. berthaultii Hawkes, aux larves de Leptinotarsa decemlineata Say, ont été étudiés par ablation sélective des trichomes glandulaires et par l'élimination de leur exsudat des folioles, et par comparaison avec S. tuberosum L. qui a perdu les trichomes glandulaires défensifs A et B. L'ablation des trichomes A a augmenté la proportion de larves ayant consommé S. berthaultii. L'élimination de l'exsudat des trichomes B a augmenté la proportion de consommatrices et réduit la mortalité. Les principaux composés actifs de l'exsudat B, c'est-à-dire des esters d'acides gras de sucrose, n'étaient actifs qu'en présence de trichomes A. Les esters de sucrose n'ont pas modifié la consommation larvaire sur folioles de S. tuberosum, ou sur disques de feuilles de S. berthaultii dont les trichomes A avaient été enlevés. La croissance des larves survivantes n'a pas été modifiée significativement par l'ablation des trichomes A ou l'élimination de l'exsudat de B. La croissance des larves a été significativement augmentée quand les folioles de S. berthaultii ont été incorporés dans l'aliment artificiel après élimination de la barrière physique due aux pédoncules B. La croissance a été de même importance sur aliments artificiels contenant des feuilles (fraiches ou en poudre lyophylisée) de S. berthaultii ou de S. tuberosum, mais plus faible que sur folioles de S. tuberosum. La présence de trichomes A est indispensable à la résistance de S. berthaultii aux L, de L. decemlineata. Les gouttelettes de type B contenant des esters de sucrose augmentent l'expression de la résistance en présence d'une défense active par trichomes A.

     

Alpha-momorcharin enhances Nicotiana benthamiana resistance to tobacco mosaic virus infection through modulation of reactive oxygen species

Alpha-momorcharin (α-MMC), a member of the plant ribosomal inactivating proteins (RIPs) family, has been proven to exhibit important biological properties in animals, including antiviral, antimicrobial, and antitumour activities. However, the mechanism by which α-MMC increases plant resistance to viral infections remains unclear. To study the effect of α-MMC on plant viral defence and how α-MMC increases plant resistance to viruses, recombinant DNA and transgenic technologies were employed to investigate the role of α-MMC in Nicotiana benthamiana resistance to tobacco mosaic virus (TMV) infection. Treatment with α-MMC produced through DNA recombinant technology or overexpression of α-MMC mediated by transgenic technology alleviated TMV-induced oxidative damage and reduced the accumulation of reactive oxygen species (ROS) during TMV-green fluorescent protein infection of N. benthamiana. There was a significant decrease in TMV replication in the upper leaves following local α-MMC treatment and in α-MMC-overexpressing plants relative to control plants. These results suggest that application or overexpression of α-MMC in N. benthamiana increases resistance to TMV infection. Finally, our results showed that overexpression of α-MMC up-regulated the expression of ROS scavenging-related genes. α-MMC confers resistance to TMV infection by means of modulating ROS homeostasis through controlling the expression of antioxidant enzyme-encoding genes. Overall, our study revealed a new crosstalk mechanism between α-MMC and ROS during resistance to viral infection and provides a framework to understand the molecular mechanisms of α-MMC in plant defence against viral pathogens.     

Wild species of Nicotiana as a new source of tobacco resistance to the tobacco hornworm Manduca sexta

Sixteen species of Nicotiana were examined for resistance to second instar larvae of the tobacco hornworm, Manduca sexta (L.). The tests were designed so as to discriminate between antibiosis and nonpreference. High levels of antibiosis resistance were observed in several wild species such as N. gossei and species in the Repandae section. Several other species, such as N. excelsior, showed antibiosis that appeared to be due to a different mechanism than the alkaloid-trichome exudate based resistance of the above species. The data indicate that these species of Nicotiana may be used as a new source of resistance to larvae of M. sexta.

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D'espèces sauvages de Nicotiania comme nouvelle source de résistance du tabac à Manduca sexta

Résumé La résistance aux chenilles de second stade de Manduca sexta L. a été examinée chez seize espèces de Nicotiana. Les essais ont été conçus pour distinguer absence d'appétance et antibiose. Plusieurs espèces sauvages comme N. gossei et les espèces de la section Repandae ont présenté un niveau élevé d'antibiose. D'autres espèces, comme N. excelsior ont présenté une antibiose vraisemblablement due à un mécanisme différent de la résistance des espèces précédentes provoquée par un exsudat alcaloïde des trichomes. Les résultats indiquent que ces espèces de Nicotiana peuvent être utilisées comme nouvelle source de résistance aux chenilles de M. sexta.

     

Hooked trichomes and resistance of Phaseolus vulgaris to Empoasca fabae (Harris)

Two cultivars of the bean, Phaseolus vulgaris L., differing in densities of hooked trichomes were examined for resistance to the potato leafhopper, Empoasca fabae. The major plant factor affecting leafhopper performance was the density of hooked trichomes. Leafhoppers were impaled by these epidermal appendages, leading to wounding and death. Nymphal survival on the cultivar, California Light Red Kidney (hooked trichome density: 2000/cm²) was significantly less than that on Brasil 343 (hooked trichome density: 400/cm²). Hooked trichomes limit damage to beans by reducing the population density of the potato leafhopper.

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Zusammenfassung Hakenhaare der Bohne, Phaseolus vulgaris L. schützen gegen Befall durch die Kartoffelzikade, Empoasca fabae (Harris). Zikadenlarven werden auf diesen Haaren aufgespiesst. Sie werden verwundet, bleiben hängen und sterben vorzeitig. Zwei Bohnensorten mit unterschiedlicher Dichte an Hakenhaaren wurden auf ihren Einfluss auf Eiablage und Entwicklungsbiologie eingezwingerter Zikaden geprüft. Eiablage, Entwicklung und Wachstum unterschieden sich auf beiden Sorten nicht. Dagegen war die Larvenmortalität, welche durch das Aufspiessen an Haaren verursacht wird, auf der resistenten Sorte California Light Red Kidney 4–10 mal grösser als auf der anfälligen Sorte Brasil 343, welche eine fünfmal geringere Dichte an Hakenhaaren hat.Im Zeltversuch wurden Populationen von Imagines und von Larven auf der Sorte California Light Red Kidney um 66%, beziehungsweise 88% reduziert verglichen mit solchen auf Brasil 343. Das Wachstum zikadenbefallener und zikadenfreier Pflanzen beider Sorten wurde ebenfalls im Zeltversuch untersucht. Die Reduktion der Blattfläche und des Trockengewichts von Blättern, Stengeln und Wurzeln war bei zikadenbefallenen Brasil 343 signifikant grösser als bei der resistenten Sorte. Demgemäss schliessen wir, dass Hakenhaare für sich allein wirksam sind, den Zikadenschaden bei California Light Red Kidney zu reduzieren und zwar durch Begrenzung der Überlebensrate und der Endpopulation.

     

Uncoupling resistance from cell death in the hypersensitive response of Nicotiana species to cauliflower mosaic virus infection

Cauliflower mosaic virus strain W260 elicits a hypersensitive response (HR) in leaves of Nicotiana edwardsonii, an interspecific hybrid derived from a cross between N. glutinosa and N. clevelandii. Interestingly, we found that N. glutinosa is resistant to W260, but responds with local chlorotic lesions rather than necrotic lesions. In contrast, N. clevelandii responds to W260 with systemic cell death. The reactions of the progenitors of N. edwardsonii to W260 infection indicated that each contributed a factor toward the development of HR. In this study, we present two lines of evidence to show that the resistance and cell death that comprise the HR elicited by W260 can indeed be uncoupled. First, we showed that the non-necrotic resistance response of N. glutinosa could be converted to HR when these plants were crossed with N. clevelandii. Second, we found that cell death and resistance segregated independently in the F2 population of a cross between N. edwardsonii and N. clevelandii. We concluded that the resistance of N. edwardsonii to W260 infection was conditioned by a gene derived from N. glutinosa, whereas cell death was conditioned by a gene derived from N. clevelandii. An analysis of pathogenesis-related (PR) protein expression in response to W260 infection revealed that elicitation of PR proteins was associated with resistance rather than with the onset of cell death.     

Enhancement of resistance to aphids by introducing the snowdrop lectin genegna into maize plants

In order to enhance the resistance to pests, transgenic maize (Zea mays L.) plants from elite inbred lines containing the gene encoding snowdrop lectin (Galanthus nivalis L. agglutinin; GNA) under control of a phloem-specific promoter were generated through theAgrobacterium tumefaciens- mediated method. The toxicity of GNA-expressing plants to aphids has also been studied. The independently derived plants were subjected to molecular analyses. Polymerase chain reaction (PCR) and Southern blot analyses confirmed that thegna gene was integrated into maize genome and inherited to the following generations. The typical Mendelian patterns of inheritance occurred in most cases. The level of GNA expression at 0.13%-0.28% of total soluble protein was observed in different transgenic plants. The progeny of nine GNA-expressing independent transformants that were derived separately from the elite inbred lines DH4866, DH9942, and 8902, were selected for examination of resistance to aphids. These plants synthesized GNA at levels above 0.22% total soluble protein, and enhanced resistance to aphids was demonstrated by exposing the plants to corn leaf aphid (Rhopalosiphum maidis Fitch) under greenhouse conditions. The nymph production was significantly reduced by 46.9% on GNA-expressing plants. Field evaluation of the transgenic plants supported the results from the inoculation trial. After a series of artificial self-crosses, some homozygous transgenic maize lines expressing GNA were obtained. In the present study, we have obtained new insect-resistant maize material for further breeding work.     

Laboratory assessments of resistance to the aphids Sitobion avenae and Metopolophium dirhodum in three Triticum species and two modern wheat cultivars

Three ancient varieties of wheat and two spring-sown modern varieties were screened in the laboratory to assess their resistance to the cereal aphids Sitobion avenae and Metopolophium dirhodum. Resistance was measured in terms of non-preference and antibiosis on plants at three growth stages. The ancient variety Einkorn was the most resistant in terms of both non-preference and antibiosis to both species of aphids at all growth stages examined. The ancient variety Emmer and the modern Sicco exhibited some resistant properties, whereas the ancient Spelt and modern Timmo were relatively susceptible to aphid attack.     

Role of trichomes in soybean resistance to cabbage looper, Trichoplusia ni

Role of leaf trichome density and length in Glycine max (L.) Merr. resistance to Trichoplusia ni (Hübner) was evaluated at different leaf positions on a relatively resistant soybean, plant introduction (PI) 227687, and a relatively susceptible cultivar, Davis. The uppermost (juvenile) leaf within both PI 227687 and Davis plants, which is densely covered with trichomes, was most resistant to T. ni larval feeding and survival. When these trichomes were shaven off, such leaves became as susceptible to T. ni larval feeding as unshaven TL3 and TL5 leaves (PI 227687) or all other unshaven leaves (Davis). Bioassays for antifeedants in ethyl acetate and hexane extractables from leaves at the different positions on PI 227687 and Davis plants showed that the resistance observed in the uppermost Davis leaf is attributable to trichomes (i.e., a morphological factor); whereas, in the uppermost PI 227687 leaf it involves both morphological (trichomes) and chemical factors, but the trichome parameter is dominant.

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Résumé L'influence de la densité et de la longueur des trichomes de Glycine max sur la résistance à Trichoplusia ni a été évaluée suivant la position des feuilles sur une lignée résistante (PI 227687) et sur un cultivar sensible (Davis).Les feuilles apicales (juvéniles) tant de PI 227687 que de Davis, recouvertes de trichomes denses, résistèrent mieux à l'alimentation larvaire et à la survie de T. ni.Quand ces trichomes étaient éliminés, ces feuilles ne résistaient pas plus à l'alimentation des larves de T. ni que les feuilles non rasées TL3 et TL5 de PI 227687 ou toutes les autres feuilles de Davis.Les tests avec des extraits dans l'acétate d'éthyle et l'hexane de feuilles provenant de différentes positions de PI 227687 et Davis, destinés à mettre en évidence des phagodissuadants, ont montré que la résistance observée chez les feuilles apicales de Davis était attribuable aux trichomes (c'est-à-dire à un caractère morphologique), tandis que chez les mêmes feuilles de PI 227687 elle impliquait à la fois des trichomes (morphologie) et des substances chimiques, mais avec une prédominance de l'influence des trichomes.

     

The evolutionary ecology of symbiont-conferred resistance to parasitoids in aphids

Abstract Aphids may harbor a wide variety of facultative bacterial endosymbionts. These symbionts are transmitted maternally with high fidelity and they show horizontal trans-mission as well, albeit at rates too low to enable infectious spread. Such symbionts need to provide a net fitness benefit to their hosts to persist and spread. Several symbionts haveachieved this by evolving the ability to protect their hosts against parasitoids. Reviewing empirical work and some models, I explore the evolutionary ecology of symbiont-conferredresistance to parasitoids in order to understand how defensive symbiont frequencies are maintained at the intermediate levels observed in aphid populations. I further show thatdefensive symbionts alter the reciprocal selection between aphids and parasitoids by augmenting the heritable variation for resistance, by increasing the genetic specificity of thehost-parasitoid interaction, and by inducing environment-dependent trade-offs. These effects are conducive to very dynamic, symbiont-mediated coevolution that is driven by frequency-dependent selection. Finally I argue that defensive symbionts represent a problem for biological control of pest aphids, and I propose to mitigate this problem byexploiting the parasitoids＇ demonstrated ability to rapidly evolve counteradaptations to symbiont-conferred resistance.